A pyrazolo pyridine derivative and use thereof

Abstract

The present invention is in the field of medicinal chemistry and relates to a novel pyrazolopyridine derivative and its preparation and use as therapeutic agents, in particular as TRK inhibitors. Compounds of general formula (I) and their geometric isomers or pharmaceutically acceptable salts thereof and their preparation. Preferred compounds have activity as protein kinase inhibitors, in particular tropomyosin receptor kinase (TRK) inhibitors.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry and relates to a novel pyrazolopyridine derivative, its preparation method, and its application as a therapeutic agent, particularly as a TRK inhibitor. Background Technology

[0002] For a long time, people have suffered from malignant tumors without a perfect cure. The fight against cancer has gradually shifted from the use of traditional non-selective chemotherapy drugs to highly selective targeted therapies. Since the successful launch of the first kinase inhibitor, imatinib, and its promising clinical results, research on kinase inhibitors has continued to deepen.

[0003] TRK, short for tropomyosin receptor kinase, belongs to the transmembrane protein kinase family. It consists of a ligand-binding extracellular domain, a kinase-active intracellular domain, and a transmembrane domain. There are three subtypes of TRK: TRKA, TRKB, and TRKC, each binding to different extracellular ligands to exert its biological activity. TRKA primarily binds to nerve growth factor (NGF), TRKB primarily binds to brain-derived neurotrophic factor (BDNF), and TRKC primarily binds to neurotrophin-3 (NT-3). The intracellular kinase domain then undergoes continuous phosphorylation and autophosphorylation, activating downstream signaling pathways and exerting biological activities, including inducing cell proliferation and differentiation, and inhibiting apoptosis. Continuous activation of TRK may lead to cancer. NTRK is the DNA sequence encoding TRK. NTRK has three homologs: NTRK1, NTRK2, and NTRK3, corresponding to the three TRK subtypes. NTRK gene fusion is a major pathogenic factor in TRK-related cancers. NTRK gene fusion refers to the linking of the 3' end of the NTRK gene encoding the TRK protein with the 5' end of a gene encoding other unrelated proteins. This results in the loss of the extracellular domain of the synthesized TRK protein, while the intracellular kinase domain remains continuously activated, potentially leading to cancer. Researchers have observed NTRK gene fusion in various tumors, such as the TPM3-NTRK1 fusion in colorectal cancer (CRC) and the ETV6-NTRK3 fusion in secretory mesodermal fibrosarcoma and infantile fibrosarcoma. Due to the loss of the extracellular domain, monoclonal antibody inhibitors are ineffective. Therefore, small molecule inhibitors are an important therapeutic approach targeting TRK-related cancers. Summary of the Invention

[0004] The purpose of this invention is to provide a novel pyrazolopyridine derivative, its preparation method, and its use as a therapeutic agent, particularly in TRK inhibitors.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] A pyrazolopyridine derivative, wherein the derivative is a compound represented by general formula I, and its geometric isomers, enantiomers, or pharmaceutically acceptable salts thereof; the compound represented by general formula I is as follows:

[0007]

[0008] Among them, L1 and L2 are independently selected from (NH) and may be identical or different. n (CH2) n where n = 0 or 1;

[0009] L3 is selected from (C=O) m m = 0 or 1;

[0010] Ring A is selected from unsubstituted or aryl groups substituted by 1-5 identical or different Ra groups;

[0011] The B ring is selected from unsubstituted aryl groups, wherein L2 and L3 are in the adjacent, meta, or para positions of the B ring;

[0012] The C ring is selected from unsubstituted or substituted rings with at least one R ring. b Substituted C3-C6 aliphatic rings containing 1-2 heteroatoms;

[0013] Ra is selected from hydrogen, halogen, nitro or C1-C4 haloalkyl;

[0014] R b Selected from C1-C6 alkyl, hydroxyl, or amino groups.

[0015] Preferably, the derivative is a compound represented by general formula I, its geometric isomers, enantiomers, or pharmaceutically acceptable salts thereof;

[0016] Among them, L1 and L2 are independently selected from (NH) and may be identical or different. n (CH2) n where n = 0 or 1;

[0017] L3 is selected from (C=O) m m = 0 or 1;

[0018] Ring A is selected from unsubstituted or aryl groups substituted by 1-5 identical or different Ra groups;

[0019] The B ring is selected from unsubstituted aryl groups, wherein L2 and L3 are in the adjacent, meta, or para positions of the B ring;

[0020] The C ring is selected from unsubstituted or substituted rings with at least one R ring. b Substituted C3-C5 aliphatic rings containing 1-2 heteroatoms;

[0021] Ra is selected from hydrogen, halogen, nitro or C1-C2 haloalkyl;

[0022] R b Selected from C1-C3 alkyl, hydroxyl, or amino groups.

[0023] More preferably, the derivative is a compound represented by general formula I, and its geometric isomers, enantiomers, or pharmaceutically acceptable salts thereof;

[0024] Among them, L1 and L2 are independently selected from (NH) and may be identical or different. n (CH2) n where n = 0 or 1;

[0025] L3 is selected from (C=O) m m = 0 or 1;

[0026] Ring A is selected from unsubstituted or aryl groups substituted by 1-5 identical or different Ra groups;

[0027] Ring B is selected from benzene ring, wherein L2 and L3 are in the ortho, meta, or para position of ring B;

[0028] The C ring is selected from morpholino, N-methylpiperazinyl, piperazinyl, piperidine-4-amino, 4-aminopiperidinyl, (R / S)-3-hydroxypiperidinyl, and 4-hydroxypiperidinyl.

[0029] Ra is selected from hydrogen, fluorine, chlorine, bromine, nitro or C1-C2 haloalkyl.

[0030] Further preferred, the derivative is a compound represented by general formula I, and its geometric isomers, enantiomers, or pharmaceutically acceptable salts thereof;

[0031] L1 is selected from methylene;

[0032] L2 is selected from (NH) n n = 0 or 1;

[0033] L3 is selected from (C=O) m m = 0 or 1;

[0034] Ring A is selected from unsubstituted or aryl groups substituted by 1-5 identical or different Ra groups;

[0035] Ring B is selected from the benzene ring, wherein L2 and L3 are in the meta or para position of ring B;

[0036] The C ring is selected from morpholino, N-methylpiperazinyl, piperazinyl, piperidine-4-amino, 4-aminopiperidinyl, (R / S)-3-hydroxypiperidinyl, and 4-hydroxypiperidinyl.

[0037] Ra is selected from hydrogen, fluorine, chlorine, bromine, nitro or trifluoromethyl;

[0038] More preferably, the derivative is:

[0039] (3-(5-(3-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0040] (3-(5-benzyl-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0041] (3-(5-(3-bromobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0042] (3-(5-(3-chlorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0043] Morpholinyl(3-(5-(3-nitrobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl) methyl ketone;

[0044] Morpholinyl(3-(5-(3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl) methyl ketone;

[0045] (3-(5-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0046] (3-(5-(4-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0047] (3-(5-(2,3-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0048] (3-(5-(2,4-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0049] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0050] (3-(5-(2,6-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0051] (3-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0052] Morpholinyl(3-(5-(3,4,5-trifluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl) methyl ketone;

[0053] 5-(2,5-difluorobenzyl)-N-(3-morpholinophenyl)-1H-pyrazolo[3,4-b]pyridine-3-amine;

[0054] 5-(2,5-difluorobenzyl)-N-(4-morpholinophenyl)-1H-pyrazolo[3,4-b]pyridine-3-amine;

[0055] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-methylpiperazin-1-yl)methyl ketone;

[0056] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone;

[0057] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone;

[0058] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(piperazin-1-yl)methyl ketone;

[0059] (4-aminopiperidin-1-yl)(3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone;

[0060] 3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)-N-(piperidin-4-yl)benzamide;

[0061] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone;

[0062] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-methylpiperazin-1-yl)methyl ketone;

[0063] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone;

[0064] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone;

[0065] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(piperazin-1-yl)methyl ketone;

[0066] (4-aminopiperidin-1-yl)(4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone;

[0067] 4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)-N-(piperidin-4-yl)benzamide;

[0068] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-methylpiperazin-1-yl)methyl ketone;

[0069] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone;

[0070] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone;

[0071] (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(piperazin-1-yl)methyl ketone;

[0072] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-methylpiperazin-1-yl)methyl ketone;

[0073] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone;

[0074] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone;

[0075] (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(piperazin-1-yl)methyl ketone;

[0076] (R)-(3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone;

[0077] (S)-(3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone.

[0078] The use of pyrazolopyridine derivatives, compounds of general formula I, and their geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof in the preparation of medicaments for the prevention or treatment of diseases related to the expression or activity of TRK kinases.

[0079] The use of the compound represented by general formula I, its geometric isomers, or pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof in the preparation of a treatment for the prevention or treatment of tumors, cancers, or severe pain caused by the TRK pathway.

[0080] A pharmaceutical composition comprising a compound of general formula I and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs as an active ingredient and pharmaceutically acceptable excipients.

[0081] The use of the pharmaceutical composition, specifically its use in the preparation of medicaments for the prevention or treatment of diseases related to the expression or activity of TRK kinases.

[0082] The composition is used in the preparation of medicaments for the prevention or treatment of tumors, cancers, or severe pain caused by the TRK pathway.

[0083] In the definitions of compounds of general formula I given above, the general definitions used in this compilation are as follows:

[0084] Halogens: refer to fluorine, chlorine, bromine, or iodine. Substituents: such as methyl, nitro, halogen, etc. Haloalkyl groups: straight-chain or branched alkyl groups, where the hydrogen atoms on these alkyl groups may be partially or completely replaced by halogen atoms, for example, chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, etc. Substituted aliphatic rings, such as morpholino, N-methylpiperazinyl, piperazinyl, piperidin-4-amino, 4-aminopiperidinyl, (R / S)-3-hydroxypiperidinyl, 4-aminopiperidinyl, etc.

[0085] The preparation method of the derivatives shown in General Formula I above is as follows:

[0086]

[0087] (1) The target derivative has a structure as shown in general formula 10 or a similar structure, wherein Ar is 0-5 R a The substituted benzene ring structure, and R a It is a halogen, nitro, or haloalkyl group. Prepared according to the method shown in Route 1:

[0088] Step a: m-aminobenzoic acid and morpholine undergo a condensation reaction at room temperature to obtain intermediate (3);

[0089] Step b; 5-Bromo-1H-pyrazolo[3,4-b]pyridine reacts with NIS via an iodination reaction to give intermediate (5);

[0090] Step c: Intermediate (5) reacts with PMBCl under low-temperature alkaline conditions to obtain intermediate (6);

[0091] Step d: Intermediate (6) undergoes Bulchwald coupling with intermediate 3 to obtain intermediate (7);

[0092] Step e: Intermediate (7) reacts with pinacol diboronate via the Miyaura reaction to obtain intermediate (8);

[0093] Step f: Intermediate (8) reacts with benzyl bromide of different substitutions in a Suzuki reaction to give intermediate (9);

[0094] Step g: The intermediate (9) is deprotected from PMB under acidic conditions to obtain the final product (10).

[0095] The preferred conditions are as follows:

[0096] In step a, morpholine (2) (26.69 mmol) was dissolved in N,N-dimethylformamide, and HOBt (29.11 mmol), EDCI (29.11 mmol), and DIPEA (48.51 mmol) were added dropwise with stirring at room temperature. After the addition was complete, the reaction was carried out at room temperature for 2 h. The reaction solution was poured into water and extracted with ethyl acetate (250 mL × 3). The organic layer was washed with saturated ammonium chloride (100 mL × 2) and saturated brine (100 mL × 2). The solution was dried over anhydrous sodium sulfate and the solvent was evaporated to obtain intermediate (3). The condensing agent can be selected from HOBt plus EDCI, HATU, HBTU and PyBop, etc., with HOBt plus EDCI being preferred; the acid binding agent can be selected from DIPEA, TEA and DMAP, etc., with DIPEA being preferred; the solvent can be selected from tetrahydrofuran, acetonitrile, N,N-dimethylformamide and dimethyl sulfoxide, etc., with N,N-dimethylformamide being preferred.

[0097] In step b, 5-bromo-1H-pyrazolo[3,4-b]pyridine (4) (121.20 mmol) and NIS (133.00 mmol) were dissolved in N,N-dimethylformamide, and the mixture was stirred at 60 °C for 12 h. After cooling to room temperature, the reaction solution was poured into water, filtered, washed, and dried to obtain intermediate (5). The reaction solvent can be acetonitrile, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, etc., preferably N,N-dimethylformamide.

[0098] In step c, intermediate (5) (92.90 mmol) was dissolved in anhydrous N,N-dimethylformamide. Sodium hydride (111.48 mmol) was added in portions at 0°C and stirred for 2 h. PMBCl (102.19 mmol) was added at 0°C, and the mixture was transferred to room temperature and stirred for 4 h. The reaction solution was poured into ice water, and the system was adjusted to neutral with ammonium chloride. The mixture was filtered, washed with a mixture of ethyl acetate and methanol (1:1), and then recrystallized from N,N-dimethylformamide to obtain the purified intermediate (6). The reaction solvent must be anhydrous and can be acetonitrile, dimethyl sulfoxide, tetrahydrofuran, or N,N-dimethylformamide, preferably N,N-dimethylformamide. The reaction temperature is -78 to 0°C, preferably 0°C. The base in the reaction can be sodium hydride, sodium hydroxide, potassium hydroxide, etc., preferably sodium hydride.

[0099] In step d, intermediate (6) (15.76 mmol), intermediate (3) (18.91 mmol), XantPhos (1.57 mmol) and cesium carbonate (31.53 mmol) were dissolved in anhydrous 1,4-dioxane, and Pd2(dba)3 (1.57 mmol) was added under argon protection. The reaction was carried out at 90 °C for 8 h under argon atmosphere. The organic solvent was removed by evaporation, and intermediate (7) was obtained by column chromatography. The palladium complex can be Pd(PPh3)4, Pd(dppf)Cl2, Pd2(dba)3, and Pd(OAc)2, with Pd2(dba)3 being preferred. The inorganic base can be selected from potassium acetate, potassium fluoride, sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, potassium hydroxide, sodium tert-butoxide, etc., with cesium carbonate being preferred. The solvent is anhydrous and can be 1,4-dioxane, tetrahydrofuran, toluene, N,N-dimethylformamide, dimethyl sulfoxide, etc., with anhydrous 1,4-dioxane being preferred.

[0100] In step e, intermediate (7) (0.38 mmol), pinacol diborate (0.96 mmol), and potassium acetate (1.15 mmol) were dissolved in anhydrous 1,4-dioxane (10 mL), and Pd(dppf)Cl2 (0.019 mmol) was added under argon protection. The mixture was reacted at 100 °C for 4 h under argon atmosphere and then proceeded directly to the next step without further treatment. The palladium complex can be Pd(PPh3)4, Pd(dppf)Cl2, Pd2(dba)3, and Pd(OAc)2, with Pd(dppf)Cl2 being preferred. The inorganic base can be selected from potassium acetate, potassium fluoride, sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, potassium hydroxide, sodium tert-butoxide, etc., with potassium acetate being preferred. The solvent is anhydrous and can be 1,4-dioxane, tetrahydrofuran, toluene, N,N-dimethylformamide, dimethyl sulfoxide, etc., with anhydrous 1,4-dioxane being preferred.

[0101] In step f, 2 mL of water, cesium carbonate (1.15 mmol), benzyl bromo with different substitutions (1.15 mmol) and Pd(PPh3)4 (0.019 mmol) were added to the reaction solution in step e. The mixture was stirred at 80 °C for 2 h under an argon atmosphere, cooled, the solvent was evaporated, and the intermediate (9) was obtained by column chromatography. The palladium complex can be Pd(PPh3)4, Pd(dppf)Cl2, Pd2(dba)3, and Pd(OAc)2, with Pd(PPh3)4 being preferred. The inorganic base can be selected from potassium acetate, potassium fluoride, sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, potassium hydroxide, sodium tert-butoxide, etc., with cesium carbonate being preferred. The solvent can be 1,4-dioxane, tetrahydrofuran, toluene, N,N-dimethylformamide, dimethyl sulfoxide, water, and ethylene glycol dimethyl ether, etc., or it can be a mixed solvent composed of two solvents, with a mixed solvent of 1,4-dioxane and water (5:1) being preferred.

[0102] In step g, intermediate (9) was dissolved in trifluoroacetic acid (5 mL), reacted at 60 °C for 4 h with stirring, trifluoroacetic acid was removed by evaporation, the pH of the system was adjusted to alkaline by saturated sodium bicarbonate, filtered and washed, and the filter cake was washed with methanol with stirring to obtain the final product (10).

[0103]

[0104] (2) The target derivative has a structure as shown in general formula 17 or a similar structure, wherein R is 3-ethoxycarbonyl or 4-ethoxycarbonyl, R1 is 3-carboxylic acid or 4-carboxylic acid, and R2 is an unsubstituted or substituted C3-C6 aliphatic ring containing 1-2 heteroatoms linked by a carbonyl group. Prepared according to the method shown in route 2:

[0105] Step h, intermediate (6) undergoes Bulchwald coupling with ethyl 3-aminobenzoate or ethyl 4-aminobenzoate to obtain intermediate 12;

[0106] Step i, intermediate (12) reacts with pinacol diboronate via the Miyaura reaction to obtain intermediate (13);

[0107] Step j, intermediate (13) undergoes Suzuki coupling with 2,5-difluorobenzyl bromide to obtain intermediate (14);

[0108] Step k, intermediate (14) is deprotected under trifluoroacetic acid conditions to obtain intermediate (15);

[0109] Step 1: The intermediate (15) is deethylated under strongly alkaline conditions to obtain intermediate (16);

[0110] In step m, intermediate (16) undergoes a condensation reaction with different amines to obtain the final product (17). If the amine contains a tert-butoxycarbonyl group for protection, the protection is then removed in the next step.

[0111] The preferred conditions are as follows;

[0112] Step h is similar to step d, except that the solvent is replaced with N,N-dimethylformamide and the temperature is raised to 120°C.

[0113] Step i is similar to step e, except that the inorganic base is replaced by cesium carbonate instead of potassium acetate.

[0114] Step j is similar to step f, except that the reaction time is extended to 4 hours.

[0115] In step k, intermediate (14) (1.14 mmol) was dissolved in trifluoroacetic acid (10 mL), and the mixture was stirred at 60 °C for 5 h. The trifluoroacetic acid was then removed by evaporation and the process proceeded directly to the next step.

[0116] In step 1, intermediate (15) is dissolved in methanol (9 mL), water (3 mL) and sodium hydroxide (2.0 g) are added, and the mixture is reacted at 60 °C with stirring for 4 h. After cooling, the reaction solution is poured into water, washed once with ethyl acetate (20 mL), and the pH of the aqueous layer is adjusted to 4 with concentrated hydrochloric acid. The mixture is filtered, washed, and dried to obtain intermediate (16). The solvent can be a mixture of methanol, ethanol, and water, preferably a mixture of methanol and water; the strong base can be sodium hydroxide, potassium hydroxide, etc., preferably sodium hydroxide; the temperature can be 50–80 °C, preferably 60 °C.

[0117] In step m, intermediate (16) (0.11 mmol) and PyBop (0.21 mmol) were dissolved in tetrahydrofuran, and DIPEA (0.32 mmol) was added at 0 °C. The mixture was stirred for 1 h, and a tetrahydrofuran solution of the corresponding amine was added dropwise. The mixture was stirred for another 1 h. The reaction solution was poured into water and extracted with ethyl acetate (10 mL × 3). The organic phase was washed with saturated ammonium chloride (10 mL), the solvent was evaporated, and column chromatography was used to obtain the final product (17). If the amine was protected with a tert-butoxycarbonyl group, the column chromatography product was dissolved in a saturated solution of hydrogen chloride in ethyl acetate, stirred at room temperature for 0.5 h, the solvent was evaporated, the pH of the system was adjusted to 8 with saturated sodium bicarbonate, and the mixture was extracted with ethyl acetate (10 mL × 3). The product was dried over anhydrous sodium sulfate, and the solvent was evaporated to obtain the final product (17). The condensing agent can be selected from HOBt plus EDCI, HATU, HBTU, PyBop, etc., with PyBop being preferred; the solvent can be selected from dichloromethane, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, etc., with tetrahydrofuran being preferred; the reaction temperature can be selected from -40 to 20℃, with 0℃ being preferred; the acid-binding agent can be selected from DIPEA, TEA, DMAP, etc., with DIPEA being preferred.

[0118]

[0119] (3) The target derivative has a structure as shown in general formula 24 or a similar structure, wherein R is 3-ethoxycarbonyl or 4-ethoxycarbonyl, R1 is 3-carboxylic acid or 4-carboxylic acid, and R2 is an unsubstituted or substituted C3-C6 aliphatic ring containing 1-2 heteroatoms linked by a carbonyl group. Prepared according to the method shown in route 3:

[0120] In step n, intermediate (6) and intermediate (18) undergo a Suzuki reaction to obtain intermediate (19);

[0121] Step o, intermediate (19) reacts with pinacol diboronic acid ester to obtain intermediate (20);

[0122] Step p, intermediate (20) reacts with 2,5-difluorobenzyl bromide in a Suzuki reaction to give intermediate (21);

[0123] Step q: Intermediate (21) is deprotected under strong acid conditions to obtain intermediate (22);

[0124] Step r, intermediate (22) is deprotected under strong alkaline conditions to obtain intermediate (23);

[0125] In step s, the intermediate (23) is condensed with different amines to obtain the final product (24). If the amine contains a tert-butoxycarbonyl group for protection, the protection is then removed in the next step.

[0126] The preferred conditions are as follows:

[0127] In step n, intermediates (6) (6.76 mmol) and (18) (7.43 mmol) were dissolved in 1,4-dioxane (50 mL), potassium fluoride (13.5 mmol), S-Phos (1.35 mmol) were added, 10 mL of water was added, Pd(PPh3)4 (0.34 mmol) was added under an argon atmosphere, and the reaction was carried out at 100 °C for 24 h under argon protection. The solvent was removed by evaporation, and the intermediate (19) was obtained by column chromatography and stirring with methanol. The palladium complex can be Pd(PPh3)4, Pd(dppf)Cl2, Pd2(dba)3, and Pd(OAc)2, with Pd(PPh3)4 being preferred. The inorganic base can be selected from potassium acetate, potassium fluoride, sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, potassium hydroxide, sodium tert-butoxide, etc., with potassium fluoride being preferred. The solvent can be 1,4-dioxane, tetrahydrofuran, toluene, N,N-dimethylformamide, dimethyl sulfoxide, water, and ethylene glycol dimethyl ether, etc., or a mixed solvent composed of two solvents, with a mixed solvent of 1,4-dioxane and water being preferred.

[0128] Step o is similar to step e, except that the inorganic base is replaced with potassium acetate, the palladium catalyst is replaced with Pd(OAc)2, and the organophosphorus ligand XantPhos (0.2 eq) is added;

[0129] Step p is similar to step f, except that the temperature is raised to 100°C and the reaction time is extended to 7 hours;

[0130] Step q is the same as step k;

[0131] Step r is the same as step l;

[0132] Step s is the same as step m.

[0133] The use of pyrazolopyridine derivatives, compounds of general formula I, and their geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof in the preparation of medicaments for the prevention or treatment of diseases related to the expression or activity of TRK kinases.

[0134] The use of the compound represented by general formula I, its geometric isomers, or pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof in the preparation of a treatment for the prevention or treatment of tumors, cancers, or severe pain caused by the TRK pathway.

[0135] A pharmaceutical composition comprising a compound of general formula I and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs as an active ingredient and pharmaceutically acceptable excipients.

[0136] The composition is used in the preparation of medicaments for the prevention or treatment of diseases related to the expression or activity of TRK kinase.

[0137] The composition is used in the preparation of drugs for the prevention or treatment of tumors, cancers, or severe pain caused by the TRK pathway.

[0138] Advantages of this invention:

[0139] This invention focuses on NTRK gene fusion-type cancers, designs a series of pyrazolo[3,4-b]pyridine derivatives, and finds that compounds with such structures exhibit good TRK inhibitory activity, which can be used to treat cancers caused by NTRK gene fusions or other diseases related to abnormal TRK expression. Detailed implementation method:

[0140] The examples are intended to illustrate, and not limit, the scope of the invention. The 1H NMR spectra of the compounds were determined using a Bruker ARX-600 or Bruker ARX-400; all reagents used were analytically or chemically pure.

[0141] The specific implementation structure is as follows:

[0142] Example 1

[0143] The preparation route of (3-(5-(3-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone is shown below:

[0144] Synthesis of (3-aminophenyl)(morpholino)methyl ketone (3)

[0145] Morpholine (2) (26.69 mmol) was dissolved in N,N-dimethylformamide, and HOBt (29.11 mmol), EDCI (29.11 mmol), and DIPEA (48.51 mmol) were added. A solution of m-aminobenzoic acid (1) (25.00 mmol) in N,N-dimethylformamide was added dropwise with stirring at room temperature. After the addition was complete, the reaction mixture was allowed to react at room temperature for 2 h. The reaction solution was then poured into water, extracted with ethyl acetate (250 mL × 3), washed with saturated ammonium chloride (100 mL × 2) and saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, and the solvent was evaporated to obtain a brown oily substance with a yield of 82.3%.

[0146] 2) Synthesis of 5-bromo-3-iodo-1H-pyrazolo[3,4-b]pyridine (5)

[0147] 5-Bromo-1H-pyrazolo[3,4-b]pyridine (4) (121.10 mmol) and NIS (133.00 mmol) were dissolved in N,N-dimethylformamide, heated to 60 °C and stirred for 12 h, cooled to room temperature, the reaction solution was poured into water, filtered, washed and dried to give a pale yellow solid with a yield of 82.4%.

[0148] 3) Synthesis of 5-bromo-3-iodo-1-(4-methoxybenzyl)-1H-pyrazole[3,4-b]pyridine (6)

[0149] The obtained compound (5) (92.90 mmol) was dissolved in anhydrous N,N-dimethylformamide. Sodium hydride (111.48 mmol) was added in portions at 0 °C and stirred for 2 h. PMBCl (102.19 mmol) was added at 0 °C, and the mixture was transferred to room temperature and stirred for 4 h. The reaction solution was poured into ice water, and the system was adjusted to neutral with ammonium chloride. The mixture was filtered, and the filter cake was washed with ethyl acetate and methanol (1:1) with stirring. The mixture was then recrystallized from N,N-dimethylformamide to obtain a white solid with a yield of 50.2%.

[0150] Synthesis of (4)(3-(5-bromo-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (7)

[0151] Intermediate (6) (15.76 mmol), intermediate (3) (18.91 mmol), XantPhos (1.57 mmol), and cesium carbonate (31.53 mmol) were dissolved in anhydrous 1,4-dioxane. Pd2(dba)3 (1.57 mmol) was added under argon protection, and the reaction was carried out at 90 °C for 8 h under argon atmosphere. The organic solvent was removed by evaporation, and the solid was purified by column chromatography to obtain a yellow solid with a yield of 48.5%.

[0152] Synthesis of (5)(3-(1-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (8)

[0153] Intermediate (7) (0.38 mmol), pinacol diboronate (0.96 mmol), and potassium acetate (0.96 mmol) were dissolved in anhydrous 1,4-dioxane (10 mL). Pd(dppf)Cl2 (0.019 mmol) was added under argon protection, and the mixture was reacted at 100 °C for 4 h under argon atmosphere. The mixture was then proceeded directly to the next step without further treatment.

[0154] Synthesis of (6)(3-(5-(3-fluorobenzyl)-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (9)

[0155] Add 2 mL of water, cesium carbonate (1.15 mmol), 3-fluorobenzyl bromide (1.15 mmol), and Pd(PPh3)4 (0.019 mmol) to the reaction solution obtained in step 5) above. Stir the reaction at 80 °C for 2 h under an argon atmosphere, cool, evaporate the solvent, and obtain a yellow solid by column chromatography with a yield of 48.4%–53.5%.

[0156] 7. Preparation of (3-(5-(3-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (Example 1)

[0157] Intermediate (9) was dissolved in trifluoroacetic acid (5 mL) and reacted at 60 °C with stirring for 4 h. The trifluoroacetic acid was removed by evaporation, and the pH of the system was adjusted to alkaline using saturated sodium bicarbonate. The mixture was filtered and washed, and the filter cake was washed with methanol with stirring to obtain a pale yellow solid, with a yield of 49.9%. Ms(ESI)m / z 431.2, [M+H] + 432.1. 1 H NMR (400MHz, DMSO-d6) δ12.55(s,1H),9.19(s,1H),8.45(d,J=2.0Hz,1H),8.18(d,J=1.8Hz,1H),7.80(t,J=3.2,J=1.6,1H),7.61(dd,J=8, J=1.6,1H),7.30-7.39(m,2H),7.11(dd,J=8.8,J=1.2,2H),7.06-7.01(m,1H),6.83(dt,J=7.4,1.3Hz,1H),4.11(s,2H),3.60-3.57(m,8H). 13 C NMR (151MHz, DMSO-d6) δ169.94,163.61,161.99,151.25,151.00,144.79(d,J=7.6Hz),143.81,143.06,136.57,130.98(d,J=8.8Hz),129.29 (d,J=3.3Hz),127.68,125.26(d,J=3.2Hz),117.93,117.31,115.87(d,J=20.5Hz),114.69,113.44(d,J=21.3Hz),106.94,66.68(2C),38.06.

[0158] Example 2

[0159] Preparation of (3-(5-benzyl-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone (Example 2)

[0160] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with benzyl bromide in an equal proportion to obtain Example 2. Ms(ESI)m / z 413.2, [M+H] + 414.2. 1 H NMR(600MHz,DMSO-d6)δ12.70(s,0.2H),12.54(s,0.8H),9.44(s,0.2H),9.25(s,0.8H),8.80(s,0.2H),8.75(s,0.2H), 8.43(s,0.8H),8.21(s,0.8H),7.89(s,0.2H),7.81(s,0.8H),7.75-7.68(m,0.8H),7.62(d,J=8.3Hz,0.8H),7.54-7.51 (m,0.4H),7.41-7.39(m,0.2H),7.36(d,J=7.8Hz,0.2H),7.33(s,0.4H),7.31(s,0.8H),7.30(s,0.8H),7.28(s,0.8H), 7.26(s,0.8H),7.20(t,J=7.3Hz,1H),6.87(d,J=7.8Hz,0.2H),6.83(d,J=7.5Hz,0.8H),4.23-4.08(m,2H),3.60(s,8H). 13 C NMR(151MHz,DMSO-d6)δ169.95,151.21,151.01,143.80,143.09,141.87,136.54,129.64,129.1 1(2C),129.08(2C),128.26,127.19,126.62,117.89,117.30,114.67,106.94,66.68(2C),38.54.

[0161] Example 3

[0162] Preparation of (3-(5-(3-bromobenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl) methyl ketone (Example 3)

[0163] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with m-bromobenzyl bromide in equal proportion, thus obtaining Example 3. Ms(ESI)m / z 492.0, [M+H] + 493.0. 1H NMR (600MHz, DMSO-d6) δ12.57(s,1H),9.23(s,1H),8.45(d,J=1.9Hz,1H),8.17(d,J=1.7Hz,1H),7.81(t,J=1.8,1H),7.61(dd,J=7.8,J=1.2, 1H),7.50(s,1H),7.41(dq,J=5.6,3.6,2.8Hz,1H),7.32(t,J=7.8Hz,1 H),7.30-7.23(m,2H),6.83(d,J=7.8Hz,1H),4.10(s,2H),3.62(m,8H). 13 C NMR(151MHz,DMSO-d6)δ169.94,151.25,151.00,144.78,143.81,143.05,136.56,131.81,131.26,129.5 5,129.31(2C),128.29,127.64,122.36,117.93,117.32,114.70,106.94,66.68(2C),65.39(2C),37.91.

[0164] Example 4

[0165] Preparation of (3-(5-(3-chlorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone (Example 4)

[0166] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 3-chlorobenzyl bromide in equal proportion, resulting in Example 4. Ms(ESI)m / z 447.1, [M+H] + 448.1. 1 H NMR (600MHz, DMSO-d6) δ12.58(s,1H),9.23(s,1H),8.46(d,J=1.8,1H),8.18(s,1H),7.81(s,1H),7.62(d,J=7.9Hz,1H ),7.36-7.31(m,3H),7.28(d,J=7.7Hz,1H),7.25(d,J=7.2,1H),6.84(d,J=7.2Hz,1H),4.11(s,2H),3.62-3.39(m,8H). 13C NMR(151MHz,DMSO-d6)δ169.93,151.25,151.01,144.47,143.81,143.05,136.57,133.65,130.9 5,129.30,128.95,127.90,127.64,126.66,117.93,117.31,114.70,106.94,66.68(2C),37.94.

[0167] Example 5

[0168] Preparation of morpholino(3-(5-(3-nitrobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl) methyl ketone (Example 5)

[0169] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 3-(nitro)benzyl bromide in proportion, thus obtaining Example 5. Ms(ESI)m / z 458.2, [M+H] + 459.1. 1 HNMR (600MHz, DMSO-d6) δ12.60(s,1H),9.25(s,1H),8.50(d,J=1.8Hz,1H),8.22(d,J=2.2Hz,1H),8.16(t,J=2.0Hz,1H),8.09(dd,J=8.4 ,J=1.8,1H),7.81(s,1H),7.76(d,J=7.7Hz,1H),7.64-7.60(m,2H),7.32(t,J=7.8Hz,1H),6.83(m,1H),4.26(s,2H),3.62-3.38(m,8H). 13 C NMR(151MHz,DMSO-d6)δ169.93,151.27,151.06,148.44,144.23,143.84,143.04,136.56,136.08,1 30.62,129.52,129.29,127.23,123.66,121.78,117.94,117.32,114.70,106.97,66.68(2C),37.74.

[0170] Example 6

[0171] Preparation of morpholino(3-(5-(3-(trifluoromethyl)benzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)phenyl) methyl ketone (Example 6)

[0172] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 3-(trifluoromethyl)benzyl bromide in equal proportion, thus obtaining Example 6. Ms(ESI)m / z 481.2, [M+H] + 482.1. 1 H NMR (600MHz, DMSO-d6) δ12.59(s,1H),9.25(s,1H),8.48(d,J=2.0Hz,1H),8.20(s,1H),7.81(t,J=1.9Hz,1H), 7.66(s,1H),7.62–7.55(m,4H),7.32(t,J=7.8Hz,1H),6.83(d,J=7.8Hz,1H),4.21(s,2H),3.66-3.40(m,8H). 13 C NMR (151MHz, DMSO-d6) δ169.93,151.25,151.06,143.82,143.40,143.04,136.56,133.38,130.16(2C),129.86(d,J= 31.2Hz), 129.40 (d, J = 16.2Hz), 127.51, 125.56 (m), 123.60 (m), 117.93, 117.31, 114.70, 106.95, 66.68 (2C), 37.98.

[0173] Example 7

[0174] Preparation of (3-(5-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl) methyl ketone (Example 7)

[0175] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 2-fluorobenzyl bromide in equal proportion, resulting in Example 7. Ms(ESI)m / z 431.2, [M+H] + 432.1. 1 H NMR (600MHz, DMSO-d6) δ12.56(s,1H),9.24(s,1H),8.44(d,J=2.1Hz,1H),8.18(d,J=2.1Hz,1H),7.81(t,J=1.9Hz,1H),7.62(dd,J=8.2 ,2.3Hz,1H),7.37(td,J=7.7,1.9Hz,1H),7.33-7.28(m,2H),7.20-7.16(m,2H),6.83(d,J=7.8Hz,1H),4.11(s,2H),3.66-3.61(m,8H). 13C NMR (151MHz, DMSO-d6) δ169.94,161.64,160.02,151.23,150.85,143.77,143.06,136.55,131.64(d,J=4.4Hz),129.28,129.04( d,J=10.0Hz),128.45(d,J=15.6Hz),126.89,125.15,117.91,117.32,115.87(d,J=21.8Hz),114.71,106.85,66.68(2C),31.82.

[0176] Example 8

[0177] Preparation of (3-(5-(4-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone (Example 8)

[0178] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 4-fluorobenzyl bromide in proportion, resulting in Example 8. Ms(ESI)m / z 431.2, [M+H] + 432.2. 1 H NMR (600MHz, DMSO-d6) δ12.56(s,1H),9.21(s,1H),8.44(d,J=1.8,1H),8.15(s,1H),7.81(s,1H),7.61(dd ,J=8.1,2.3Hz,1H),7.33-7.29(m,3H),7.16-7.13(m,2H),6.83(d,J=7.2Hz,1H),4.08(s,2H),3.61(s,8H). 13 C NMR(151MHz,DMSO-d6)δ169.93,162.08,160.48,151.22,150.99,143.78,143.07,138.00(d,J=2.8Hz),136.56,130 .95(d,J=7.9Hz,2C),129.30,129.16,128.19,117.91,117.30,115.84(d,J=21.3Hz,2C),114.67,66.68(2C),37.56.

[0179] Example 9

[0180] Preparation of (3-(5-(2,3-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (Example 9)

[0181] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 2,3-difluorobenzyl bromide in equal proportion, resulting in Example 9. Ms(ESI)m / z 449.2, [M+H] + 450.1. 1 H NMR (600MHz, DMSO-d6) δ12.59(s,1H),9.24(s,1H),8.47(d,J=2.1Hz,1H),8.18(d,J=2.1Hz,1H),7.82(t,J= 1.9Hz,1H),7.62(ddd,J=8.3,2.4,1.0Hz,1H),7.34-7.30(m,2H),7.20-7.18(m,2H),6.84(dt,J=7.8,1.8Hz 1H),4.17(s,2H),3.51(m,8H). 13 C NMR (151MHz, DMSO) δ169.93, 151.23, 151.16 (dd, J = 245.3, 12.4Hz) 150.83, 149.36,(dd,J=245.3,12.4Hz),143.81,143.00,136.56,131.15(d,J=12.4H z),129.29,129.14,126.76(t,J=3.3Hz),126.32,125.48(dd,J=6.7,4.6Hz) ,117.96,117.34,116.19(d,J=16.8Hz),114.73,106.85,66.68(2C),31.53.

[0182] Example 10

[0183] Preparation of (3-(5-(2,4-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (Example 10)

[0184] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 2,4-difluorobenzyl bromide in equal proportion, resulting in Example 10. Ms(ESI)m / z 449.2, [M+H] + 450.1. 1HNMR(600MHz,DMSO-d6)δ12.57(s,1H),9.23(s,1H),8.44(s,1H),8.15(s,1H),7.81(s,1H),7.62(d,J=8.2Hz,1H),7.43(q ,J=7.8Hz,1H),7.32(t,J=7.8Hz,1H),7.26-7.22(m,1H),7.09-7.06(m,1H),6.84(m,1H),4.09(s,2H),3.66-3.62(m,8H). 13 C NMR(151MHz,DMSO-d6)δ169.94,162.45(dd,J=128.9,12.4Hz),160.83(dd,J=13 1.0,12.3Hz),151.22,150.83,143.79,143.03,136.56,132.63(dd,J=9.8,6.1H z),129.29,128.99,126.73,124.81(dd,J=15.7,3.9Hz),117.94,117.33,114.7 3,112.17(dd,J=21.0,3.7Hz),106.83,104.60(t,J=26.0Hz),66.68(2C),31.26.

[0185] Example 11

[0186] Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone (Example 11)

[0187] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 2,5-difluorobenzyl bromide in equal proportion, resulting in Example 11. Ms(ESI)m / z 449.2, [M+H] + 450.1. 1 H NMR (600MHz, DMSO-d6) δ12.58(s,1H),9.24(s,1H),8.46(d,J=2.1Hz,1H),8.18(d,J=2.1Hz,1H),7.82(t,J=1.9Hz,1H),7.62(dd,J= 8.3, 2.4Hz, 1H), 7.32 (t, J = 7.8Hz, 1H), 7.29-7.23 (m, 2H), 7.16-7.12 (m, 1H), 6.83 (d, J = 7.2Hz, 1H), 4.10 (s, 2H), 3.66-3.39 (m, 8H). 13CNMR(151MHz,DMSO-d6)δ169.93,159.43,157.84(d,J=5.2Hz),156.22,151.23,150.84,143.81,143.01,136.56,130.59(dd,J=18.3,7.9Hz), 129.29,129.10,126.32,118.01(m),117.85(dd,J=58.9,6.8Hz),117.48(m),115.41(dd,J=23.8,8.9Hz),114.73,106.84,66.68(2C),31.82.

[0188] Example 12

[0189] Preparation of (3-(5-(2,6-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (Example 12)

[0190] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 2,6-difluorobenzyl bromide in equal proportion, resulting in Example 12. Ms(ESI)m / z 449.2, [M+H] + 450.1. 1 H NMR(600MHz,DMSO-d6)δ12.87(m,0.23H),12.59(s,0.77H),9.43(m,0.15H),9.29(s,0.77H),9.26(m,0.08H),8.82(m,0.08H),8.74(m,0.15H), 8.54(d,J=2.4Hz,0.08H),8.44(d,J=2.1Hz,0.77H),8.30(d,J=2.7Hz,0 .08H),8.19(d,J=2.1Hz,1H),8.14(m,0.08H),7.90(m,0.08H),7.85(m,1 H),7.76(m,0.17H),7.70(m,0.08H),7.65(m,1H),7.56(m,0.23H),7.38 (tt,J=8.1,6.5Hz,1H),7.32(t,J=7.8Hz,0.77H),7.22(m,0.23H),7.15 (t,J=7.8Hz,1.54H),6.88(d,J=6.96Hz,0.23H),6.84(dt,J=7.5,1.3Hz ,0.77H),5.33(m,0.08H),5.19(s,0.15H),4.12(s,1.54H),3.51(m,8H).

[0191] Example 13

[0192] Preparation of (3-(5-(3,5-difluorobenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)phenyl)(morpholinyl)methyl ketone (Example 13)

[0193] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 3,5-difluorobenzyl bromide in equal proportion, resulting in Example 13. Ms(ESI)m / z 449.2, [M+H] + 450.1. 1 HNMR (400MHz, DMSO-d6) δ12.57(s,1H),9.21(s,1H),8.46(d,J=2.1Hz,1H),8.19(d,J=2.1Hz,1H),7.81(t,J=1.9Hz,1H),7.62(dd,J=8. 2,2.3Hz,1H),7.32(t,J=7.9Hz,1H),7.10-7.05(m,1H),7.05-7.00(m,2H),6.83(dt,J=7.5,1.2Hz,1H),4.12(s,2H),3.60-3.44(m,8H). 13 C NMR(151MHz,DMSO-d6)δ169.94,163.82(d,J=13.5Hz),162.18(d,J=12.7Hz),151.24,150.94,146.55,146.49(t,J=9.0Hz),143.03,136.56,12 9.42(d,J=17.8Hz),127.08,117.95,117.33,114.70,112.39(dd,J=19. 7,4.9Hz),106.97,102.37(t,J=25.9Hz),66.68(2C),55.38(2C),37.89.

[0194] Example 14

[0195] Preparation of morpholino(3-(5-(3,4,5-trifluorobenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)phenyl) methyl ketone (Example 14)

[0196] Referring to the preparation method of Example 1, the 3-fluorobenzyl bromide raw material in step 6) was replaced with 3,4,5-trifluorobenzyl bromide in equal proportion, thus obtaining Example 14. Ms(ESI)m / z 467.2, [M+H] + 468.1. 1HNMR(600MHz,DMSO-d6)δ12.60(s,1H),9.20(s,1H),8.46(s,1H),8.15(s,1H),7.81(s,1H),7.61(d,J=8.2H z,1H),7.33(t,J=7.8Hz,1H),7.27(t,J=7.8Hz,2H),6.84(d,J=7.8Hz,1H),4.10(s,2H),3.66-3.61(m,8H). 13 C NMR(151MHz,DMSO-d6)δ169.93,151.53(dd,J=9.9,4.0Hz),151.28,150.95,149.89(d d,J=10.1,3.2Hz),143.83,143.04,139.26(m),138.74(dt,J=247.0,15.6Hz),138.64 ,138.54,137.11,137.00,136.90,136.58,129.31(2C),127.00,117.95,117.32,114. 70,113.80(d,J=4.0Hz),113.69(d,J=4.0Hz),106.94,66.68(2C),55.77(2C),37.43.

[0197] Example 15

[0198] Preparation of 5-(2,5-difluorobenzyl)-N-(3-morpholinophenyl)-1H-pyrazolo[3,4-b]pyridine-3-amine (Example 15) Referring to the preparation method of Example 1, in step 4), intermediate (3) was replaced with 3-(4-morpholino)aniline in equal proportion, and the solvent was replaced with N,N-dimethylformamide. The temperature was then raised to 120°C to obtain Example 15. Ms(ESI)m / z 421.2, [M+H] + 422.2. 1 H NMR (600MHz, DMSO-d6) δ12.48(s,1H),8.91(s,1H),8.44(d,J=2.0Hz,1H),8.17(d,J=2.1Hz,1H),7.32(t,J=1.9Hz,1H),7.28–7.23 (m,2H),7.14(td,J=8.8,8.4,3.8Hz,1H),7.10–7.08(m,2H),6.45-6.42(m,1H),4.09(s,2H),3.75-3.74(m,4H),3.08–3.07(m,4H).

[0199] Example 16

[0200] Preparation of 5-(2,5-difluorobenzyl)-N-(4-morpholinophenyl)-1H-pyrazolo[3,4-b]pyridine-3-amine (Example 16) Referring to the preparation method of Example 1, in step 4), intermediate (3) was replaced with 4-(4-morpholino)aniline in equal proportion, and the solvent was replaced with N,N-dimethylformamide. The temperature was then raised to 120°C to obtain Example 16. Ms(ESI)m / z 421.2, [M+H] + 422.2. 1 H NMR (600MHz, DMSO-d6) δ12.33(s,1H),8.78(s,1H),8.42(s,1H),8.15(s,1H),7.56(d,J=8.5Hz,2H),7.25(m,2H), 7.14(dq,J=8.7,4.7,4.1Hz,1H),6.89(d,J=8.5Hz,2H),4.08(s,2H),3.73(t,J=4.8Hz,4H),2.99(t,J=4.7Hz,4H).

[0201] Example 17

[0202] The preparation route of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-methylpiperazin-1-yl)methyl ketone is shown below:

[0203]

[0204] 1) Synthesis of ethyl 3-(5-bromo-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)benzoate (12)

[0205] Following the synthesis method of intermediate (7) in Example 1, intermediate (3) was replaced with ethyl 3-aminobenzoate (11), the solvent was replaced with N,N-dimethylformamide, and the temperature was raised to 120°C. A yellow solid was obtained, with a yield of 65.9%.

[0206] 2) Synthesis of ethyl benzoate (13) of 3-(1-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)benzoate

[0207] The synthesis method is the same as that of intermediate (8) in Example 1.

[0208] 3) Synthesis of ethyl benzoate (14) of 3-(5-(2,5-difluorobenzyl)-1-(4-methoxybenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)benzoate

[0209] The synthesis method of intermediate (9) in Example 1 was followed, with 3-fluorobenzyl bromide replaced by 2,5-difluorobenzyl bromide in equal proportion. 4) Synthesis of ethyl 3-(5-(2,5-difluorobenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)aminobenzoate (15)

[0210] Intermediate (14) (1.14 mmol) was dissolved in 10 mL of trifluoroacetic acid and reacted at 60 °C with stirring for 5 h. The solvent was then evaporated, and the next step was carried out directly.

[0211] 5) Synthesis of 3-(5-(2,5-difluorobenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)benzoic acid (16)

[0212] The product from step 4) was dissolved in a mixture of 9 mL methanol and 3 mL water. 2.0 g sodium hydroxide was added, and the mixture was stirred at 60 °C for 4 h. The reaction solution was poured into water and washed once with 10 mL ethyl acetate. The pH of the aqueous layer was adjusted to 4 with concentrated hydrochloric acid. The solution was extracted, washed, and dried to obtain a brown solid with a yield of 65.7%.

[0213] 6) Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-methylpiperazin-1-yl)methyl ketone (Example 17)

[0214] Intermediate (16) (40 mg, 0.11 mmol) and PyBop (0.21 mmol) were dissolved in 2 mL of tetrahydrofuran. DIPEA (0.32 mmol) was added at 0 °C and the mixture was stirred for 1 h. Then, 1 mL of a tetrahydrofuran solution of N-methylpiperazine (0.21 mmol) was added dropwise at 0 °C, and the reaction was continued for another 1 h. The reaction mixture was poured into water and extracted with ethyl acetate (10 mL × 3). The organic layer was washed with saturated ammonium chloride (10 mL). The organic phase was then subjected to column chromatography to give a white solid with a yield of 52.4%. Ms(ESI) m / z 462.2, [M+H] + 463.1. 1HNMR(600MHz,DMSO-d6)δ12.58(s,1H),9.22(s,1H),8.46(d,J=2.1Hz,1H),8.18( d,J=2.1Hz,1H),7.77(t,J=2.0Hz,1H),7.63(dd,J=8.2,2.3Hz,1H),7.31(t,J=7. 8Hz,1H),7.29-7.23(m,2H),7.13(ddd,J=8.7,6.4,3.6Hz,1H),6.80(dt,J=7.4,1 .3Hz,1H),4.10(s,2H),3.61(s,2H),3.33(s,2H),2.35-2.28(m,4H),2.19(s,3H). 13 C NMR (151MHz, DMSO-d6) δ169.81, 159.43 (d, J = 240.6Hz), 157.80 (d, J = 240.0Hz), 151.24, 150.83, 143.82, 142.99, 136.99, 130.5 9(dd,J=18.3,7.9Hz),129.28,129.11,126.32,118.00(dd,J=24.5,5.1Hz) 117.81,117.46(dd,J=25.2,8.8Hz),117.18,115.39 115.28(dd,J=24.2,8.5Hz),114.58,106.86,55.38-54.83(m,2C),49.08(2C),46.07,31.83.

[0215] Example 18

[0216] Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone (Example 18)

[0217] Referring to the preparation method of Example 17, N-methylpiperazine in step 6) was replaced with 4-hydroxypiperidine in equal proportion, resulting in Example 18. Ms(ESI) m / z 463.2, [M+H] + 464.1. 1HNMR(600MHz,DMSO-d6)δ12.57(s,1H),9.23(s,1H),8.46(d,J=1.8,1H),8.19 (s,1H),7.76(s,1H),7.63(d,J=7.7Hz,1H),7.32-7.23(m,3H),7.14(tt,J=8. 3,3.6Hz,1H),6.79(d,J=7.8Hz,1H),4.80(d,J=3.7Hz,1H),4.10(s,2H),4.02 (s,1H),3.75-3.73(m,1H),3.55(s,1H),3.20-3.16(m,2H),2.02-1.70(m,4H). 13 C NMR (151MHz, DMSO-d6) δ169.77, 159.42 (d, J = 240.2Hz), 157.80 (d, J = 240.7Hz), 151.22,150.82,143.84,142.96,137.44,130.60(dd,J=18.3,7.9Hz),129.26,1 29.14,126.29,118.01(dd,J=24.2,4.8Hz),117.54,117.47(dd,J=24.9,8.9Hz) ,117.01,115.40(dd,J=23.8,8.9Hz),114.31,106.85,66.04,45.12(2C),31.82.

[0218] Example 19

[0219] Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone (Example 19)

[0220] Referring to the preparation method of Example 17, N-methylpiperazine in step 6) was replaced with 3-hydroxypiperidine in equal proportion, resulting in Example 19. Ms(ESI) m / z 463.2, [M+H] + 464.1. 1HNMR (600MHz, DMSO-d6) δ12.57(s,1H),9.25(s,1H),8.45(s,1H),8.21(s,1H),8.16(d,J=8.5Hz,1H),7.99(s,1H),7.92(d,J=8.1Hz,1H),7.3 2(t,J=7.9Hz,1H),7.27-7.24(m,2H),7.15-7.12(m,1H),4.10(s,2H), 3.82-3.78(m,1H),3.00(m,4H),2.96-2.94(m,1H),1.73-1.71(m,4H). 13 C NMR(151MHz,DMSO-d6)δ170.16,159.43(d,J=240.0Hz),157.82(d,J=243.6Hz),151.23,1 50.81,143.85,142.96,137.52,130.60(dd,J=18.4,7.7Hz),129.15(2C),126.30,118.00( dd,J=24.3,4.9Hz),117.59,117.46(dd,J=24.9,9.0Hz),116.97,115.39(dd,J=24.1,8.6 Hz),114.51(d,J=45.9Hz),106.87,65.74-65.58(m),49.13,47.64,31.82,27.29,23.94..

[0221] Example 20

[0222] Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(piperazin-1-yl)methyl ketone (Example 20)

[0223] Following the preparation method of Example 17, N-methylpiperazine in step 6) was replaced proportionally with tert-butyl piperazine-1-carboxylate. The product obtained by column chromatography was then dissolved in a saturated solution of hydrogen chloride and ethyl acetate, stirred at room temperature for 1 h, the solvent was evaporated, the pH of the system was adjusted to 8 with saturated sodium bicarbonate, extracted with ethyl acetate (10 mL × 3), washed with saturated brine (10 mL × 3), and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration under reduced pressure, and ethyl acetate was removed by rotation to obtain a white solid. Ms(ESI) m / z 448.2, [M+H] + 449.1. 1H NMR (600MHz, DMSO-d6) δ12.58(s,1H),9.24(s,1H),8.46(d,J=2.1Hz,1H),8.19(d,J=2.1Hz, 1H),7.77(t,J=1.9Hz,1H),7.62(dd,J=8.2,2.3Hz,1H),7.31(t,J=7.8Hz,1H),7.28-7.23(m 2H),7.13(tt,J=8.2,3.6Hz,1H),6.79(d,J=7.8Hz,1H),4.10(s,2H),3.54(s,2H),3.28-3.26(m,2H),2.72-2.64(m,4H),2.43(s,1H). 13 C NMR(151MHz,DMSO-d6)δ169.79,159.42(d,J=240.7Hz),157.80(d,J=240.8Hz),15 1.23,150.82,143.84,142.97,137.25,130.60(dd,J=18.9,7.9Hz),129.25,129.1 5,126.30,118.00(dd,J=24.3,5.1Hz),117.77,117.46(dd,J=24.9,9.0Hz),117.0 5,115.39(dd,J=23.9,8.2Hz),114.54,106.86,46.51,46.34,46.31,46.03,31.82.

[0224] Example 21

[0225] Preparation of (4-aminopiperidin-1-yl)(3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone (Example 21)

[0226] Referring to Example 20, piperazine-1-carboxylic acid tert-butyl ester in step 6) was replaced proportionally with 4-tert-butoxycarbonylaminopiperidine, resulting in Example 21. Ms(ESI) m / z 462.2, [M+H] + 463.2. 1H NMR (600MHz, DMSO-d6) δ12.58(s,1H),9.22(s,1H),8.46(s,1H),8.18(s,1H),7.76(s,1H),7.62(d,J=8.2Hz,1H),7. 31-7.25(m,3H),7.14(s,1H),6.78(d,J=7.5Hz,1H),4.28(s,1H),4.10(s,2H),3.61-3.33(m,4H),1.99-1.66(m,4H).

[0227] Example 22

[0228] Preparation of 3-(5-(2,5-difluorobenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)-N-(piperidin-4-yl)benzamide (Example 22)

[0229] Referring to Example 20, piperazine-1-carboxylic acid tert-butyl ester in step 6) was replaced proportionally with 1-tert-butoxycarbonyl-4-aminopiperidine, resulting in Example 22. Ms(ESI) m / z 462.2, [M+H] + 463.1. 1 H NMR (600MHz, DMSO-d6) δ12.57(s,1H),9.25(s,1H),8.45(s,1H),8.21(s,1H),8.16(d,J=8.5Hz,1H),7.99(s,1H),7.92(d,J= 8.1Hz,1H),7.32(t,J=7.8Hz,1H),7.27-7.23(m,3H),7.15-7.12(m,1H),4.10(s,2H),3.81(s,1H),3.01(s,4H),1.72(s,4H). 13 C NMR(151MHz,DMSO-d6)δ166.47,159.43(d,J=239.4Hz),157.81(d,J=240.5Hz),151 .27,150.74,143.95,142.96,136.26,130.60(dd,J=18.8,7.7Hz),129.20,128.96,1 26.24,118.41,118.11,118.00(dd,J=24.4,5.0Hz),117.46(dd,J=24.9,9.0Hz),11 5.91,115.39(dd,J=23.9,8.3Hz),106.91,46.34,46.31,45.82,31.8,26.41,26.36.

[0230] Example 23

[0231] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(morpholino)methyl ketone (Example 23)

[0232] Referring to the preparation method of Example 17, ethyl 3-aminobenzoate in step 1) was replaced with ethyl 4-aminobenzoate in equal proportion, and N-methylpiperazine in step 6) was replaced with morpholine in equal proportion, thus obtaining Example 23. Ms(ESI) m / z 449.2, [M+H] + 450.1. 1 H NMR(600MHz,DMSO-d6)δ12.65(s,1H),9.36(s,0.87H),8.99(s,0.13H),8.70(s ,0.13H),8.47(s,0.87H),8.20(s,0.87H),8.12(s,0.13H),7.83(d,J=8.4Hz,0. 26H),7.69(d,J=8.3Hz,1.74H),7.44(d,J=7.8Hz,0.26H),7.37(d,J=8.3Hz,1. 74H),7.26(m,2H),7.13(tt,J=8.2,3.5Hz,1H),4.11(s,2H),3.60-3.51(m,8H).

[0233] Example 24

[0234] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-methylpiperazin-1-yl)methyl ketone (Example 24)

[0235] Referring to the preparation method of Example 23, morpholine in step 6) was replaced with N-methylpiperazine in proportion, resulting in Example 24. Ms(ESI)m / z 462.2, [M+H] + 463.2. 1 H NMR (600MHz, DMSO-d6) δ12.64(s,1H),9.33(s,1H),8.47(d,J=2.1Hz,1H),8.19(d,J=2.1Hz,1H),7.68(d,J=8.3Hz,2H), 7.34(d,J=8.5Hz,2H),7.29-7.23(m,2H),7.16-7.12(m,1H),4.11(s,2H),3.51(s,4H),2.33-2.31(m,4H),2.20(s,3H). 13C NMR(151MHz,DMSO-d6)δ169.84,159.43(d,J=240.0Hz),157.80(d,J=240.1Hz),151.23 ,150.86,144.30,143.56,130.59(dd,J=18.8,7.9Hz),129.15,129.04(2C),126.41,126 .29,118.01(dd,J=24.3,4.9Hz),117.47(dd,J=24.8,9.0Hz),115.48(2C),115.40(dd, J=23.9,8.9Hz),115.34,115.24,115.18,106.92,55.07(2C),49.07(2C),46.08,31.82.

[0236] Example 25

[0237] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone (Example 25)

[0238] Referring to the preparation method of Example 23, morpholine in step 6) was replaced with 4-hydroxypiperidine in an equal proportion to obtain Example 25. Ms(ESI) m / z 463.2, [M+H] + 464.1. 1 H NMR (600MHz, DMSO-d6) δ12.63(s,1H),9.31(s,1H),8.47(d,J=2.2Hz,1H),8.19(d,J=2.0Hz,1H),7.67(m,2H),7.33(m,2H),7.26 (m,2H),7.14(m,1H),4.77(d,J=4.0Hz,1H),4.11(s,2H),3.82-3.70(m,2H),3.19-3.15(m,2H),1.74(s,2H),1.38-1.23(m,3H).

[0239] Example 26

[0240] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone (Example 26)

[0241] Referring to the preparation method of Example 23, morpholine in step 6) was replaced with 3-hydroxypiperidine in an equal proportion, resulting in Example 26. Ms(ESI)m / z 463.2, [M+H]+ 464.2. 1 H NMR(600MHz,DMSO-d6)δ12.75(s,0.25H),12.62(s,0.75H),9.41(s,0.25H),9.30(s,0.75H),8.81(s,0.25H),8.71(s ,0.25H),8.47(s,0.75H),8.19(s,0.75H),7,74(m,0.5H),7.67(d,J=8.4Hz,1.5H),7.53(t,J=7.5Hz,0.5H),7.42(d,J =7.2Hz,0.25H),7.38(d,J=8.2Hz,0.5H),7.33(d,J=8.1Hz,1.5H),7.29-7.24(m,1.5H),7.13(dq,J=8.2,3.9,3.5Hz, 0.75H),4.90(s,1H),4.11(s,2H),3.81-3.48(m,3H),3.09-2.88(m,2H),1.87(s,1H),1.70(S,1H),1.41-1.38(m,2H).

[0242] Example 27

[0243] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(piperazin-1-yl)methyl ketone (Example 27)

[0244] Following the preparation method of Example 23, morpholine in step 6) was replaced proportionally with piperazine-1-carboxylic acid tert-butyl ester. The product obtained by column chromatography was then dissolved in a saturated solution of hydrogen chloride and ethyl acetate, stirred at room temperature for 1 h, and the solvent was evaporated. The pH of the system was adjusted to 8 with saturated sodium bicarbonate, extracted with ethyl acetate (10 mL × 3), washed with saturated brine (10 mL × 3), and dried over anhydrous sodium sulfate. Sodium sulfate was removed by vacuum filtration, and ethyl acetate was removed by rotary evaporation to give a pale yellow solid with a yield of 42.6%. Ms(ESI) m / z 448.2, [M+H] + 449.4. 1 H NMR(600MHz,DMSO-d6)δ12.63(s,1H),9.31(s,1H),8.47(s,1H),8.19(s,1H),7.67(d,J=8.2Hz,2H), 7.32(d,J=8.0Hz,2H),7.26(m,2H),7.14(m,1H),4.11(s,2H),3.43(s,4H),2.68(s,4H),1.23(s,1H). 13C NMR (151MHz, DMSO-d6) δ169.82, 159.42 (d, J = 240.7Hz), 157.81 (d, J = 242.1Hz), 151.23, 150.84, 144.16, 143.59, 130.59 (dd, J = 18.6, 7.9Hz), 129.1 5,128.98,126.59,126.39,118.00(dd,J=23.9,3.6Hz),117.46(dd,J=24. 9,9.1Hz),115.48,115.40(dd,J=24.5,9.4Hz),106.92,46.27(2C),31.82.

[0245] Example 28

[0246] Preparation of (4-aminopiperidin-1-yl)(4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone (Example 28)

[0247] Referring to the preparation method of Example 27, piperazine-1-carboxylic acid tert-butyl ester in step 6) was replaced with 4-tert-butoxycarbonylaminopiperidine in equal proportion, thus obtaining Example 28. Ms(ESI)m / z 462.2, [M+H] + 463.2. 1 H NMR(600MHz,DMSO-d6)δ12.63(s,1H),9.33(s,0.8H),8.99(s,0.2H),8.69(s,0.2H),8.46( s,0.8H),8.20(s,0.8H),8.12(s,0.2H),7.80(d,J=8.4Hz,0.4H),7.67(d,J=8.2Hz,1.6H), 7.37(d,J=8.4Hz,0.4H),7.31(d,J=8.3Hz,1.6H),7.25(m,2H),7.13(tt,J=8.2,3.5Hz,1H) ,4.11(s,2H),3.33(s,2H),2.97(s,2H),2.82-2.77(m,1H),1.71(s,2H),1.19–1.14(m,2H).

[0248] Example 29

[0249] Preparation of 4-(5-(2,5-difluorobenzyl)-1H-pyrazol[3,4-b]pyridin-3-yl)amino)-N-(piperidin-4-yl)benzamide (Example 29)

[0250] Referring to the preparation method of Example 27, piperazine-1-carboxylic acid tert-butyl ester in step 6) was replaced with 1-tert-butoxycarbonyl-4-aminopiperidine in equal proportion, thus obtaining Example 29. Ms(ESI)m / z 462.2, [M+H] + 463.1. 1 H NMR(600MHz,DMSO-d6)δ12.78(s,0.2H),12.66(s,0.8H),9.51(s,0.2H),9.40(s,0.8H),8.82(s,0.2H),8.73(s,0.2H ),8.47(s,0.8H),8.21(s,0.8H),7.97(d,J=7.8Hz,0.4H),7.85(d,J=8.4Hz,0.4H),7.80(d,J=8.3Hz,2H),7.75-7.72( m,1H),7.67(d,J=8.3Hz,1.6H),7.54-7.52(m,0.4H),7.42-7.39(m,0.2H),7.29–7.24(m,1.6H),7.15–7.12(m,0.8H) ,4.11(s,2H),3.81(s,1H),3.33(s,2H),2.95(d,J=11.9Hz,2H),1.99(m,0.5H),1.72-1.70(m,2H),1.43–1.38(m,2H).

[0251] Example 30

[0252] The preparation route of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-methylpiperazin-1-yl)methyl ketone is shown below.

[0253]

[0254] 1) Synthesis of ethyl 3-(5-bromo-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)benzoate (19)

[0255] Intermediate (6) (6.76 mmol) and 3-ethoxycarbonylphenylboronic acid (18) (7.43 mmol) were dissolved in 1,4-dioxane (50 mL), potassium fluoride (13.5 mmol) and S-Phos (1.35 mmol) were added, followed by 10 mL of water. Pd(PPh3)4 (0.34 mmol) was added under an argon atmosphere, and the reaction was carried out at 100 °C for 24 h under argon protection. The solvent was removed by evaporation, and the mixture was separated by column chromatography and washed with methanol to obtain a white solid with a yield of 53.7%.

[0256] 2) Synthesis of ethyl 3-(1-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl)benzoate (20)

[0257] Referring to the synthesis of intermediate (8) in Example 1, the base was replaced with an equal proportion of potassium acetate, the catalyst was replaced with an equal proportion of palladium acetate, and 0.2 equivalents of XantPhos were added.

[0258] 3) Synthesis of ethyl 3-(5-(2,5-difluorobenzyl)-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)benzoate (21)

[0259] Referring to the synthesis of intermediate (9) in Example 1, 3-fluorobenzyl bromide was replaced by 2,5-difluorobenzyl bromide in equal proportion.

[0260] 4) Synthesis of ethyl 3-(5-(2,5-difluorobenzyl)-1H-pyrazole[3,4-b]pyridin-3-yl)benzoate (22)

[0261] The synthesis is the same as that of intermediate (15) in Example 16.

[0262] 5) Synthesis of 3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)benzoic acid (23)

[0263] The synthesis is the same as that of intermediate (16) in Example 18.

[0264] 6. Synthesis of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-methylpiperazin-1-yl)methyl ketone (Example 30)

[0265] Referring to step 6) of Example 17, a white solid was obtained with a yield of 60.7%. Ms(ESI) m / z 447.2, [M+H] + 448.2. 1 H NMR (600MHz, DMSO-d6) δ13.89(s,1H),8.48(m,2H),8.08(d,J=7.8Hz,1H),7.93(s,1H),7.61(t,J=7.7Hz,1H),7.43 (d,J=7.6Hz,1H),7.27-7.21(m,2H),7.12-7.09(m,1H),4.18(s,2H),3.66(s,4H),2.42-2.32(m,4H),2.22(s,3H). 13C NMR(151MHz,DMSO-d6)δ169.11,159.43(d,J=240.0Hz),157.67(d,J=239.9Hz),1 52.48,150.56,142.02,137.19,133.79,130.59(dd,J=18.2,7.9Hz),130.01,129 .75,128.89,127.93,127.03,125.00,117.85(dd,J=23.9,4.6Hz),117.38(dd,J= 24.9, 9.0Hz), 115.36 (dd, J = 23.9, 8.9Hz), 112.35, 46.17 (2C), 45.97 (3C), 31.79.

[0266] Example 31

[0267] Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone (Example 31)

[0268] Referring to the preparation method of Example 30, N-methylpiperazine in step 6) was replaced with 4-hydroxypiperidine in proportion, resulting in Example 31. Ms(ESI) m / z 448.2, [M+H] + 449.1. 1 H NMR (600MHz, DMSO-d6) δ13.88(s,1H),8.51(d,J=2.0Hz,1H),8.44(d,J=2.0Hz,1H),8.07(dt,J=7.8,1 .4Hz,1H),7.92(t,J=1.7Hz,1H),7.61(t,J=7.7Hz,1H),7.42(dt,J=7.6,1.4Hz,1H),7.28(ddd,J=9.1 ,5.9,3.2Hz,1H),7.23(td,J=9.2,4.6Hz,1H),7.13-7.09(m,1H),4.80(d,J=4.0Hz,1H),4.18(s,2H), 4.05(s,1H),3.78-3.74(m,1H),3.56(s,1H),3.26-3.16(m,2H),1.83-1.71(m,2H),1.44-1.35(m,2H). 13C NMR (151MHz, DMSO-d6) δ169.05, 159.43 (d, J = 240.6Hz), 157.69 (d, J = 240.2Hz), 152.46,150.54,142.08,137.63,133.77,130.58(dd,J=18.2,7.9Hz),129.99,1 29.73,128.91,127.78,126.78,124.75,117.89(dd,J=24.4,4.9Hz),117.40(dd ,J=25.2,8.7Hz),115.37(dd,J=24.2,8.5Hz),112.35,65.9,46.22(2C),31.80.

[0269] Example 32

[0270] Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone (Example 32)

[0271] Referring to the preparation method of Example 30, N-methylpiperazine in step 6) was replaced with 3-hydroxypiperidine in proportion, resulting in Example 32. Ms(ESI) m / z 448.2, [M+H] + 449.0. 1 H NMR (600MHz, DMSO-d6) δ13.87(s,1H),8.51(s,1H),8.45(s,1H),8.06(d,J=7.7Hz, 1H),7.93(d,J=13.5Hz,1H),7.60(t,J=7.7Hz,1H),7.42(s,1H),7.28(ddd,J=9.0,5 .8,3.3Hz,1H),7.23(td,J=9.2,4.5Hz,1H),7.13-7.09(m,1H),5.01-4.85(m,1H), 4.18(s,2H),3.82(s,1H),3.57-3.42(m,2H),3.31-2.87(m,2H),1.90-1.40(m,4H). 13C NMR (151MHz, DMSO-d6) δ169.52 (m), 159.43 (d, J = 240.1Hz), 157.68 (d, J = 240.2Hz), 152.46, 150.52, 142.12, 137.71,133.70,130.59(dd,J=18.3,7.9Hz),130.00,129.65,128.91,127.70,127.13(m),125.18(m),117.8 9(dd,J=24.3,4.9Hz),117.38(dd,J=24.9,9.0Hz),115.36(dd,J=24.2,8.5Hz),112.36,65.50,54.41(0.5C) ,49.20(0.5C),47.73(0.5C),46.20(0.5C),33.43(0.5C),32.96(0.5C),31.81,23.87(0.5C),22.29(0.5C).

[0272] Example 33

[0273] Preparation of (3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(piperazin-1-yl)methyl ketone (Example 33)

[0274] Following the preparation method of Example 30, N-methylpiperazine in step 6) was replaced proportionally with tert-butyl piperazine-1-carboxylate. The resulting product was then dissolved in a saturated solution of hydrogen chloride in ethyl acetate, stirred at room temperature for 1 h, the solvent was evaporated, the pH of the system was adjusted to 8 with saturated sodium bicarbonate, extracted with ethyl acetate (10 mL × 3), washed with saturated brine (10 mL × 3), and dried over anhydrous sodium sulfate. Sodium sulfate was removed by vacuum filtration, and ethyl acetate was removed by rotary evaporation to give a white solid with a yield of 59.5%. Ms(ESI) m / z 433.2, [M+H] + 434.1. 1 H NMR(600MHz,DMSO-d6)δ13.89(s,1H),8.52(d,J=2.0Hz,1H),8.44(d,J=2.0 Hz,1H),8.07(d,J=7.8Hz,1H),7.92(d,J=1.9Hz,1H),7.61(t,J=7.7Hz,1H), 7.42(d,J=7.6Hz,1H),7.28(ddd,J=9.1,5.9,3.2Hz,1H),7.24(dt,J=9.2,4 .6Hz,1H),7.12(m,1H),4.19(s,2H),3.59(s,4H),2.71(m,4H),2.00(m,1H).13 C NMR(151MHz,DMSO-d6)δ169.08,159.43(d,J=239.5Hz),157.68(d,J=240.0Hz),1 52.47,150.54,142.03,137.45,133.78,130.58(dd,J=18.8,8.1Hz),129.97,129 .71,128.92,127.78,126.99,124.97,117.89(dd,J=24.7,4.6Hz),117.39(dd,J= 24.9, 9.0Hz), 115.38 (dd, J = 23.9, 8.8Hz), 112.34, 65.50 (2C), 46.34 (2C), 31.80.

[0275] Example 34

[0276] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-methylpiperazin-1-yl)methyl ketone (Example 34)

[0277] Referring to the preparation method of Example 30, 3-ethoxycarbonylphenylboronic acid in step 1) was replaced with 4-ethoxycarbonylphenylboronic acid in proportion, while N-methylpiperazine in step 6) remained unchanged, thus obtaining Example 35. Ms(ESI) m / z 447.2, [M+H] + 448.1. 1 H NMR (600MHz, DMSO-d6) δ13.90(s,1H),8.54(d,J=2.0Hz,1H),8.51(d,J=2.0Hz,1H),8.08(d,J=8.0Hz,2H),7.55(d,J=8.0Hz,2H),7.28( ddd,J=9.2,5.9,3.2Hz,1H),7.24(td,J=9.3,4.6Hz,1H),7.13-7.09(m,1H),4.18(s,2H),3.64-3.42(m,4H),2.37(m,4H),2.23(s,3H). 13C NMR (151MHz, DMSO-d6) δ169.14, 159.43 (d, J = 240.1Hz), 157.66 (d, J = 240.3Hz), 15 2.49,150.54,142.00,135.82,134.73,130.62(dd,J=18.2,7.8Hz),130.20,128.9 3,128.21(2C),126.80(2C),117.85(dd,J=24.3,5.0Hz),117.38(dd,J=24.9,9.0H z),115.35(dd,J=24.1,8.6Hz),112.44,49.20(2C),46.00(2C),31.82,27.30(3C).

[0278] Example 35

[0279] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone (Example 36)

[0280] Referring to the preparation method of Example 34, N-methylpiperazine in step 6) was replaced with 4-hydroxypiperidine in equal proportion, resulting in Example 35. Ms(ESI) m / z 448.2, [M+H] + 449.1. 1 H NMR(600MHz,DMSO-d6)δ13.89(s,1H),8.53(d,J=2.0Hz,1H),8.50(d,J=1.9Hz,1H),8 .07(d,J=7.9Hz,2H),7.53(d,J=7.9Hz,2H),7.27(ddd,J=9.4,5.9,3.4Hz,1H),7.22(d d,J=9.2,4.5Hz,1H),7.12-7.08(m,1H),4.81(d,J=3.6Hz,1H),4.17(s,2H),4.06-4.0 4(m,1H),3.78-3.73(m,2H),3.22-3.16(s,2H),1.81-1.73(m,2H),1.39-1.34(m,2H). 13C NMR(151MHz,DMSO-d6)δ169.11,159.43(d,J=240.0Hz),157.66(d,J=239.9H z),152.47,150.51,142.08,136.29,134.58,130.63(dd,J=18.9,7.9Hz),13 0.23,128.91,127.95(2C),126.79(2C),117.85(dd,J=24.3,4.9Hz),117.37 (dd,J=24.9,9.0Hz),115.34(dd,J=24.2,8.6Hz),112.45,65.98(2C),31.82.

[0281] Example 36

[0282] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone (Example 36)

[0283] Referring to the preparation method of Example 34, N-methylpiperazine in step 6) was replaced with 3-hydroxypiperidine in equal proportion, resulting in Example 36. Ms(ESI) m / z 448.2, [M+H] + 449.1. 1 H NMR (600MHz, DMSO-d6) δ13.88(s,1H),8.52(m,2H),8.07(d,J=7.9Hz,2H),7.54(s,2H),7.28(m,1H),7.22(dd,J=9.2,4.5Hz,1 H),7.10(m,1H),5.01-4.86(m,1H),4.18(s,2H),3.83(s,1H),3.54(s,2H),3.16-2.86(m,2H),1.86-1.78(m,2H),1.45(s,2H). 13C NMR(151MHz,DMSO-d6)δ169.50,159.43(d,J=240.3Hz),157.66(d,J=240.0H z),152.47,150.51,142.09,136.38,134.52,130.64(dd,J=18.2,7.9Hz),13 0.23,128.92,128.28,127.96,126.73,117.86(dd,J=24.3,4.9Hz),117.38( dd,J=24.9,9.0Hz),115.34(dd,J=23.9,8.8Hz),112.45,65.52(2C),31.82.

[0284] Example 37

[0285] Preparation of (4-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(piperazin-1-yl)methyl ketone (Example 37)

[0286] Following the preparation method of Example 34, N-methylpiperazine in step 6) was replaced proportionally with tert-butyl piperazine-1-carboxylate. The resulting product was then dissolved in a saturated solution of hydrogen chloride in ethyl acetate, stirred at room temperature for 1 h, the solvent was evaporated, the pH of the system was adjusted to 8 with saturated sodium bicarbonate, extracted with ethyl acetate (10 mL × 3), washed with saturated brine (10 mL × 3), and dried over anhydrous sodium sulfate. Sodium sulfate was removed by vacuum filtration, and ethyl acetate was removed by rotary evaporation to give a white solid with a yield of 58.1%. Ms(ESI) m / z 433.2, [M+H] + 434.1. 1 H NMR (600MHz, DMSO-d6) δ13.89(s,1H),8.52(m,2H),8.07(d,J=7.9Hz,2H),7.53(d,J=7.9Hz,2H),7.28-7.25(m,1H) ,7.23(td,J=9.3,4.5Hz,1H).,7.12-7.08(m,1H),4.23-4.20(m,1H),4.17(s,2H),3.57(s,4H),2.74-2.68(m,4H). 13C NMR(151MHz,DMSO-d6)δ169.11 159.42(d,J=240.2Hz),157.66(d,J=239.9Hz),152.49,150.52,142.03,136.10,134.58,132.00,130.63(dd,J=18.8,8.0Hz),130.21,129.1 4,128.91,128.17,126.78,117.84(dd,J=24.1,4.7Hz),117.37(dd,J=24.9,9.0Hz),115.34(dd,J=24.4,8.7Hz),112.44,65.50(2C),31.82.

[0287] Example 38

[0288] Preparation of (R)-(3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone (Example 38)

[0289] Referring to the preparation method of Example 30, N-methylpiperazine in step 6) was replaced with (R)-3-hydroxypiperidine, resulting in Example 38. Ms(ESI) m / z 448.2, [M+H] + 449.1. 1 H NMR (600MHz, DMSO-d6) δ13.88(s,1H),8.51(d,J=1.9Hz,1H),8.45(s,1H),8.07(d,J=7.8Hz ,1H),7.96-7.34(m,1H),7.60(t,J=7.7Hz,1H),7.43(t,J=7.3Hz,1H),7.27(ddd,J=9.1,5.9 ,3.2Hz,1H),7.22(td,J=9.2,4.5Hz,1H),7.10(tt,J=8.2,3.5Hz,1H),5.03-4.874.95(m,1H ),4.18(s,2H),3.83-3.82(m,1H),3.58-3.50(m,2H),3.28–2.85(m,2H),1.93–1.42(m,4H). 13CNMR(151MHz,DMSO-d6)δ169.55-169.31(m),158.63(d,J=240.2Hz),156.89(d,J=239.6Hz),152.47 ,150.52,142.13,137.69,133.70,130.48(dd,J=18.2,7.9Hz),129.99,129.66,128.92(2C),127.72, 117.78(dd,J=24.4,4.9Hz),117.25(dd,J=24.9,9.0Hz),115.23(dd,J=24.2,8.5Hz),112.37(2C),65 .50, 51.80 (d, J = 784.9Hz), 44.97 (d, J = 831.7Hz), 33.18 (d, J = 71.0Hz), 31.82, 23.05 (d, J = 237.4Hz).

[0290] Example 39

[0291] Preparation of (S)-(3-(5-(2,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone (Example 39)

[0292] Referring to the preparation method of Example 30, N-methylpiperazine in step 6) was replaced with (S)-3-hydroxypiperidine, resulting in Example 39. Ms(ESI) m / z 448.2, [M+H] + 449.1. 1 H NMR (600MHz, DMSO-d6) δ13.88(s,1H),8.51(d,J=2.0Hz,1H),8.45(s,1H),8.07(dt,J=7.8,1 .5Hz,1H),7.95(d,J=8.2Hz,1H),7.60(t,J=7.7Hz,1H),7.44-7.41(m,1H),7.27(ddd,J=9.1 ,5.9,3.2Hz,1H),7.22(td,J=9.2,4.6Hz,1H),.10(ddd,J=12.1,8.3,3.5Hz,1H),5.03-4.87 (m,1H),4.18(s,2H),3.82(s,1H),3.59-3.50(m,2H),3.20–2.85(m,2H),1.92–1.41(m,4H). 13CNMR(151MHz,DMSO-d6)δ169.54-169.31(m),158.63(d,J=240.5Hz),156.89(d,J=240.0Hz),152.46,1 50.51,137.70,133.71,130.49(dd,J=18.9,7.9Hz),129.99,129.64,128.91,128.29,127.72,119.58, 117.78(dd,J=23.9,4.6Hz),117.25(dd,J=24.9,9.0Hz),115.23(dd,J=24.2,8.5Hz),112.37,110.11, 65.50, 51.81 (d, J = 785.0Hz), 44.97 (d, J = 831.3Hz), 33.18 (d, J = 71.1Hz), 31.81, 23.06 (d, J = 238.6Hz).

[0293] Example 40: In vitro enzyme inhibitory activity study of some pyrazolopyridine derivatives of the present invention

[0294] Experimental materials:

[0295] Tecan F500 microplate reader.

[0296] KinEASE™-STK kit (containing biotinylated peptide substrate S2, Eu) 3+ Labeled monoclonal antibodies targeting specific phosphorylation sites, Sa-XL665-labeled streptavidin, kinase reaction buffer (KinEASE enzyme reaction buffer), 384 shallow well plates, full-length TRKA protein.

[0297] TRKA protein concentration 0.111 ng / μl, MgCl2, EDTA, DL-Dithiothreitol (DTT), DMSO.

[0298] Experimental methods:

[0299] Step 1: Kinase Response

[0300] First, the compound samples prepared in the above examples were dissolved in DMSO to form a 20 mM solution. Then, according to the testing requirements, the solutions were diluted with kinase reaction buffer to concentrations of 100 μM, 10 μM, and 1 μM. Next, TRKA kinase (0.111 ng / μL), ATP (4 μM), biotin-labeled peptide substrate TK (1 μM), and the compound sample (4 μL) were added to 10 μL of kinase reaction buffer (containing 5 mM MgCl2 and 1 mM DTT). The mixture was incubated at room temperature for 40 minutes, during which the kinase phosphorylated the substrate TK. Then, 10 μL of an EDTA-containing detection reagent (included in the kit) was added to detect the phosphorylation product.

[0301] Step 2: Detection of phosphorylation products:

[0302] rare earth element europium (Eu) 3+ The XL665-labeled antibody recognizes phosphorylated substrates, and streptavidin binds to biotin on the substrate. Eu3 + It is a fluorescent donor, and XL665 is a fluorescent acceptor. When Eu 3+ Similar to XL665, Eu 3+ Energy is transferred to the XL665 to generate an HTRF signal.

[0303] Result evaluation method: Fluorescence signal is derived from Eu 3+ The HTRF absorption signals at 620 nm and 665 nm of XL665 were generated. Therefore, the ratio of the HTRF signal (665 / 620) for each well plate reaction was calculated. The result was characterized as Delta F (DF%):

[0304]

[0305] Calculation of inhibition rate (activity %): The DF% of kinase activity without the addition of the compound sample is defined as 100%. When the compound sample is added, the kinase activity rate is:

[0306]

[0307] Computing IC 50 With the addition of the compound, the DF% of kinase activity was plotted on the Y-axis, and the logarithm of the compound concentration was plotted on the X-axis. IC 50 The values ​​were obtained by fitting the data into an S-type stoichiometric response curve.

[0308] The above test results show that all the above embodiments have a good inhibitory effect on TRKA kinase, and the IC50 of most embodiments is [missing information]. 50 The value is in the range of nM.

[0309] Table 1: IC50 of some aminopyrimidine derivatives of the present invention50 value

[0310] Example <![CDATA[IC 50 (μM)]]> Example <![CDATA[IC 50 (μM)]]> Example 1 0.294 Example 2 0.479 Example 3 4.430 Example 4 0.802 Example 5 1.400 Example 6 16.690 Example 7 0.978 Example 8 2.780 Example 9 1.966 Example 10 0.788 Example 11 0.178 Example 12 0.838 Example 13 0.759 Example 14 51.070 Example 15 0.507 Example 16 0.519 Example 17 0.118 Example 18 0.613 Example 19 0.205 Example 20 0.117 Example 21 0.437 Example 22 0.630 Example 23 0.980 Example 24 0.389 Example 25 0.129 Example 26 0.156 Example 27 0.409 Example 28 0.263 Example 29 0.321 Example 30 0.106 Example 31 0.245 Example 32 0.0560 Example 33 0.391 Example 34 0.296 Example 35 0.383 Example 36 0.571 Example 37 0.251 Example 38 0.057 Example 39 0.026

[0311] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A pyrazolopyridine derivative, characterized in that: The derivative is a compound represented by general formula I, or a pharmaceutically acceptable salt thereof; The compounds represented by general formula I are as follows: General Formula I Wherein, L1 is methylene; L2 is (NH) n , where n = 0 or 1; L3 is selected from (C=O) m m = 0 or 1; L2 and L3 are positioned between or opposite each other in ring B; Ring A is selected from unsubstituted phenyl groups or phenyl groups substituted with 1-5 identical or different Ra atoms; Ring B is selected from unsubstituted phenyl groups; The C ring is selected from morpholinoyl group, N -methylpiperazinyl, piperazinyl, piperidine-4-amino, 4-aminopiperidineyl, ( R )-3-hydroxypiperidinyl, ( S )-3-hydroxypiperidinyl, 4-hydroxypiperidinyl; Ra is selected from hydrogen, halogen, nitro or C1-C4 haloalkyl.

2. The pyrazolopyridine derivative according to claim 1, characterized in that: The derivative is a compound represented by general formula I, or a pharmaceutically acceptable salt thereof; Wherein, L1 is methylene; L2 is (NH) n , where n = 0 or 1; L3 is selected from (C=O) m m = 0 or 1; L2 and L3 are positioned between or opposite each other in ring B; Ring A is selected from unsubstituted phenyl groups or phenyl groups substituted with 1-5 identical or different Ra atoms; Ring B is selected from unsubstituted phenyl groups; The C ring is selected from morpholinoyl group, N -methylpiperazinyl, piperazinyl, piperidine-4-amino, 4-aminopiperidineyl, ( R )-3-hydroxypiperidinyl, ( S )-3-hydroxypiperidinyl, 4-hydroxypiperidinyl; Ra is selected from hydrogen, halogen, nitro or C1-C2 haloalkyl.

3. The pyrazolopyridine derivative according to claim 2, characterized in that: The derivative is a compound represented by general formula I, or a pharmaceutically acceptable salt thereof; Wherein, L1 is methylene; L2 is (NH) n , where n = 0 or 1; L3 is selected from (C=O) m m = 0 or 1; L2 and L3 are positioned between or opposite each other in ring B; Ring A is selected from unsubstituted phenyl groups or phenyl groups substituted with 1-5 identical or different Ra atoms; Ring B is selected from unsubstituted phenyl groups; The C ring is selected from morpholinoyl group, N -methylpiperazinyl, piperazinyl, piperidine-4-amino, 4-aminopiperidineyl, ( R )-3-hydroxypiperidinyl, ( S )-3-hydroxypiperidinyl, 4-hydroxypiperidinyl; Ra is selected from hydrogen, fluorine, chlorine, bromine, nitro or C1-C2 haloalkyl.

4. The pyrazolopyridine derivative according to claim 3, characterized in that: The derivative is a compound represented by general formula I, or a pharmaceutically acceptable salt thereof; L1 is selected from methylene; L2 is (NH) n , where n = 0 or 1; L3 is selected from (C=O) m m = 0 or 1; L2 and L3 are positioned between or opposite each other in ring B; Ring A is selected from unsubstituted phenyl groups or phenyl groups substituted with 1-5 identical or different Ra atoms; Ring B is selected from unsubstituted phenyl groups; The C ring is selected from morpholinoyl group, N -methylpiperazinyl, piperazinyl, piperidine-4-amino, 4-aminopiperidineyl, ( R )-3-hydroxypiperidinyl, ( S )-3-hydroxypiperidinyl, 4-hydroxypiperidinyl; Ra is selected from hydrogen, fluorine, chlorine, bromine, nitro or trifluoromethyl.

5. The pyrazolopyridine derivative according to claim 4, characterized in that: The derivative is: (3-(5-(3-fluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-benzyl-) 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(3-bromobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(3-chlorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; Morpholinyl (3-(5-(3-nitrobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone; Morpholinyl (3-(5-(3-(trifluoromethyl)benzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone; (3-(5-(2-fluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(4-fluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(2,3-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(2,4-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(2,6-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (3-(5-(3,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; Morpholinyl (3-(5-(3,4,5-trifluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone; 5-(2,5-Difluorobenzyl)-N-(3-morpholinophenyl)- 1H -Pyrazolo[3,4-b]pyridine-3-amine; 5-(2,5-Difluorobenzyl)-N-(4-morpholinophenyl)- 1H -Pyrazolo[3,4-b]pyridine-3-amine; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-methylpiperazin-1-yl)methyl ketone; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)3-hydroxypiperidin-1-yl)methyl ketone; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)piperazin-1-yl)methyl ketone; (4-aminopiperidin-1-yl)(3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone; 3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)-N-(piperidin-4-yl)benzamide; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)morpholino)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-methylpiperazin-1-yl)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)3-hydroxypiperidin-1-yl)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)piperazin-1-yl)methyl ketone; (4-Aminopiperidin-1-yl)(4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)phenyl)methyl ketone; 4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)amino)-N-(piperidin-4-yl)benzamide; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-methylpiperazin-1-yl)methyl ketone; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone; (3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(piperazin-1-yl)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-methylpiperazin-1-yl)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(4-hydroxypiperidin-1-yl)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone; (4-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(piperazin-1-yl)methyl ketone; ( R )-(3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone; ( S )-(3-(5-(2,5-difluorobenzyl)- 1H -pyrazolo[3,4-b]pyridin-3-yl)phenyl)(3-hydroxypiperidin-1-yl)methyl ketone.

6. The use of the pyrazolopyridine derivative according to any one of claims 1-5, characterized in that: The use of the compound represented by general formula I, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for the prevention or treatment of diseases related to the expression or activity of TRKA kinase.

7. The use of the pyrazolopyridine derivative according to any one of claims 1-5, characterized in that: The compound represented by general formula I, or a pharmaceutically acceptable salt thereof, is used in the preparation of a compound for the prevention or treatment of tumors or severe pain caused by the TRKA pathway.

8. A pharmaceutical composition, characterized in that: The active ingredient comprises a compound of general formula I according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

9. The use of the pharmaceutical composition according to claim 8, characterized in that: The composition is used in the preparation of medicaments for the prevention or treatment of diseases related to the expression or activity of TRKA kinase.

10. The use of the pharmaceutical composition according to claim 8, characterized in that: The composition is used in the preparation of medicaments for the prevention or treatment of tumors or severe pain caused by the TRKA pathway.

Images