CN111718351B - Oxygen-containing substituted pyrazolopyrimidine compound, pharmaceutical composition and application thereof - Google Patents

Abstract

The invention relates to the field of biomedicine, and discloses an oxygen-containing substituted pyrazolopyrimidine compound, a pharmaceutical composition and application thereof. The oxygen-containing substituted pyrazolopyrimidine compound having the structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite or a prodrug thereof shows excellent inhibitory activity on TRK kinase, and simultaneously shows good antitumor activity on an animal level.

Description

Oxygen-containing substituted pyrazolopyrimidine compound, pharmaceutical composition and application thereof

Technical Field

The invention relates to the field of biomedicine, and particularly relates to an oxygen-containing substituted pyrazolopyrimidine compound, a pharmaceutical composition containing the oxygen-containing substituted pyrazolopyrimidine compound, and application of the oxygen-containing substituted pyrazolopyrimidine compound and the pharmaceutical composition.

Background

NTRK/TRK (neurotrophin receptor kinase) is a neurotrophic factor tyrosine receptor, belonging to the receptor tyrosine kinase family. The TRK family mainly comprises 3 members, NTRK1/TRKA, NTRK2/TRKB and NTRK 3/TRKC. The complete TRK kinase comprises three parts, namely an extracellular region, a transmembrane region and an intracellular region. The extracellular region of TRK kinase can cause the change of kinase configuration after being combined with corresponding ligand to form dimer. The intracellular region of TRK kinase is autophosphorylated to activate the kinase activity of the TRK kinase, and further activate the downstream signal transduction pathway (such as MAPK, AKT, PKC and the like) to generate corresponding biological functions; wherein NGF (nerve growth factor) binds TRKA, BDNF (derived neurotrophic factor) binds TRKB, and NT3 (neurotrophic factor 3) binds TRKC.

TRK kinases play important physiological roles in the development of nerves, including the growth and functional maintenance of neuronal axons, the development of memory, and the protection of neurons from injury, among others. Meanwhile, a large number of researches show that activation of a TRK signal transduction pathway is closely related to generation and development of tumors, and activated TRK signal proteins are found in neurocytoma, prostatic cancer, breast cancer and the like.

In recent years, the discovery of various TRK fusion proteins shows that the TRK fusion proteins can promote the biological function of tumorigenesis. The earliest TPM3-TRKA fusion protein was found in colon cancer cells, with an incidence of about 1.5% in the clinical patients tested. Later, different types of TRK fusion proteins were found in different types of clinical tumor patient samples, such as lung cancer, head and neck cancer, breast cancer, thyroid cancer, glioma and the like, such as CD74-NTRK1, MPRIP-NTRK1, QKI-NTRK2, ETV6-NTRK3, BTB1-NTRK3 and the like. Under the condition that ligand binding is not needed, the different NTRK fusion proteins are in a highly activated kinase activity state, so that downstream signal pathways can be continuously phosphorylated, cell proliferation is induced, and generation and development of tumors are promoted.

Therefore, in recent years, TRK fusion proteins have become a potent anticancer target and a hotspot for research, for example, WO2010048314, WO2012116217, WO2011146336, W02010033941, WO2018077246 and the like all disclose TRK kinase inhibitors with different structural types.

In addition, target mutations occurring after continuous administration are important causes of tumor resistance, and recent clinical cases of TRK mutations, such as TRKA G595R, G667C, G667S and F589L (Russo M et al; Cancer Discovery,2016,6(1), 36-44), TRKC G623R and G696A (Drilon A. et al Annals of Oncology 2016,27(5), 920-.

In addition, nitrogen-containing aromatic heterocycles are generally preferred for their potency, a typical example being the ALK kinase inhibitor crizotinib (Cui J. et al. J. Med. chem.2011,54, 6342-. WO2007147647 and WO2007025540 also disclose pyrazole substituted pyrazolopyridine compounds and pyrazole substituted imidazopyridazine compounds, respectively, as ALK kinase inhibitors and their use in the treatment of disease.

Disclosure of Invention

An object of the present invention is to provide a novel oxygen-containing substituted pyrazolopyrimidine compound having excellent antitumor activity.

Although the typical compound a and the typical compound B provided in the prior art have good inhibitory activity on ALK kinase, the inhibitory effect of structural analogs represented by the typical compound a and the typical compound B on TRK kinase is not good. Through a large number of scientific researches, the inventor of the invention finds that the oxygen-containing substituted pyrazolopyrimidine compound having the structure shown in formula (I) of the invention has excellent inhibitory activity on TRK kinase, and the inhibitory activity is obviously superior to that of a typical compound A and a typical compound B in the prior art. More importantly, the pyrazolopyrimidine compound containing the phenoxy structure has the antitumor activity on the animal level which is obviously better than that of the typical compound A and the typical compound B, so that the pyrazolopyrimidine compound has more excellent tumor treatment effect than that of the prior art.

In order to achieve the above objects, a first aspect of the present invention provides an oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, or prodrug thereof,

wherein, in the formula (I),

R₁、R₂、R₃and R₄Each independently selected from H, halogen, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl groups of (a);

R₅selected from H, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, alkoxy-substituted C_2-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl groups of (a);

R₆selected from H, C_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl and halogen of (a);

R₇is selected from C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl of, C_2-12Acyl group and sulfonyl group of (1).

A second aspect of the present invention provides use of the oxygen-containing substituted pyrazolopyrimidine compound having the structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof, described in the first aspect, for the preparation of a medicament for the prevention and/or treatment of a TRK kinase receptor-mediated disease.

A third aspect of the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or diluent, and, as an active ingredient, an oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof, according to the first aspect of the present invention.

A fourth aspect of the invention provides the use of a pharmaceutical composition according to the third aspect of the invention in the manufacture of a medicament for the prevention and/or treatment of a TRK kinase receptor mediated disease.

A fifth aspect of the present invention provides an oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof, described in the first aspect of the present invention, or an application of a pharmaceutical composition described in the third aspect of the present invention in preparation of a medicament for preventing and/or treating tumors.

The oxygen-containing substituted pyrazolopyrimidine compound having the structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite or a prodrug thereof shows excellent inhibitory activity on TRK kinase, and simultaneously shows good antitumor activity on an animal level.

Detailed Description

The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.

As previously mentioned, the first aspect of the present invention provides an oxygen-containing substituted pyrazolopyrimidine compound or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof.

Some of the terms involved in the present invention are explained below:

“C_1-12the alkyl group of (1) represents an alkyl group having 1 to 12 carbon atoms in total, and includes a straight-chain alkyl group, a branched-chain alkyl group or a cyclic alkyl group, and may be, for example, a straight-chain alkyl group, a branched-chain alkyl group or a cyclic alkyl group having 1,2, 3, 4,5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms in total, and may be, for example, a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, an isobutyl group, a tert-butyl group, a n-pentyl group, an isopentyl group, a n-hexyl group, a cyclopropyl group, a methylcyclopropyl group, an ethylcyclopropyl group, a cyclopentyl group, a methylcyclopentyl group, a cyclohexyl group or the like. For "C_1-8Alkyl of (2), "" C_1-6The "alkyl group" has a similar explanation except that the number of carbon atoms is different.

"C substituted by 1-6 halogens_1-12The "alkyl group" of (A) represents an alkyl group having a total number of carbon atoms of 1 to 12, including a straight-chain alkyl group, a branched-chain alkyl group and a cycloalkyl group, and the C_1-121-6H in the alkyl group of (A) are substituted by halogen atoms selected from halogen, e.g. C_1-12In the alkyl group of (1), (2), (3), (4), (5) or (6) H is substituted by any one or more halogen atoms selected from fluorine, chlorine, bromine and iodine, and may be, for example, trifluoromethyl, difluoromethyl, monofluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, etc. For "C substituted by 1-6 halogen_1-8Alkyl of (2), "" C substituted by 1 to 6 halogens_1-6The "alkyl group" has a similar explanation except that the number of carbon atoms is different.

"hydroxy-substituted C_1-12The "alkyl group" of (A) represents an alkyl group having a total number of carbon atoms of 1 to 12, including a straight-chain alkyl group, a branched-chain alkyl group or a cyclic alkyl group, and the C_1-12At least one H in the alkyl group of (a) is substituted by a hydroxyl group.

"alkoxy-substituted C_2-12The alkyl group of (1) represents a group having 2 to 12 carbon atoms in total, and the structural formula of the group may be represented by-R¹OR²Wherein R is¹And R²Wherein the sum of the number of carbon atoms is 2 to 12, and R¹Directly connected with the phenoxy in the oxygen-containing substituted pyrazolopyrimidine compound with the structure shown in the formula (I) of the invention.

"cyano-substituted C_2-12The "alkyl group" of (A) represents an alkyl group having 2 to 12 carbon atoms in total, including a straight-chain alkyl group, a branched-chain alkyl group or a cyclic alkyl group, and the C_2-12At least one H in the alkyl group of (a) is substituted with a cyano group, and the carbon atoms in the "cyano" group count the total number of carbon atoms in the group.

"C containing 1-3 heteroatoms selected from N, O and S_2-12The "cycloalkyl group" of (a) represents a cycloalkyl group having 2 to 12 carbon atoms in total, and 1 to 3 atoms of the atoms forming the ring are hetero atoms selected from N, O and S, and the atoms forming the ring may have an alkyl substituent group having a carbon atom number included in the aforementioned range of the total carbon atom number. "C containing 1-3 heteroatoms selected from N, O and S_2-12The "cycloalkyl group" of (a) may be, for example, a three-membered ring, a four-membered ring, a five-membered ring, a six-membered ring, a seven-membered ring, an eight-membered ring, a nine-membered ring, a ten-membered ring, an eleven-membered ring or a twelve-membered ring, and H in the cycloalkyl group may be optionally substituted with, if substituted, at least one substituent independently selected from the group consisting of halogen, hydroxyl, nitro and mercapto.

“C_2-12The "acyl group" of (a) represents an acyl group having 2 to 12 carbon atoms in total, and may be, for example, an acetyl group, a propionyl group or the like.

The "sulfonyl group" may contain C_1-6The sulfonyl group may be, for example, -SO₂R³Is represented by, wherein R³Can be C_1-6Alkyl group of (1).

Several preferred embodiments of the oxygen-containing substituted pyrazolopyrimidine compounds of the structure of formula (I) according to the invention are provided below:

embodiment mode 1:

in the formula (I), the compound represented by the formula (I),

R₁、R₂、R₃and R₄Each independently selected from H, fluorine, chlorine, bromine, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl groups of (a);

R₅selected from H, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl groups of (a);

R₆selected from H, C_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl and halogen of (a);

R₇is selected from C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl of, C_2-8Acyl group and sulfonyl group of (1).

Embodiment mode 2:

in the formula (I), the compound represented by the formula (I),

R₁、R₂、R₃and R₄Each independently selected from H, fluorine, chlorine, bromine, C_1-6Alkyl of (2), C substituted by 1 to 4 halogens selected from fluorine, chlorine and bromine_1-6Alkyl groups of (a);

R₅selected from H, C_1-6Alkyl of (2), C substituted by 1 to 4 halogens selected from fluorine, chlorine and bromine_1-6Alkyl, hydroxy-substituted C of_1-6Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-6Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-8Cycloalkyl groups of (a);

R₆selected from H, C_1-6Alkyl, hydroxy-substituted C of_1-6Alkyl and halogen of (a);

R₇is selected from C_1-6Alkyl of (2), C substituted by 1 to 4 halogens selected from fluorine, chlorine and bromine_1-6Alkyl, hydroxy-substituted C of_1-6Alkyl, cyano-substituted C_2-6Alkyl group containing 1-C with 3 hetero atoms selected from N, O and S_2-8Cycloalkyl of, C_2-6Acyl group and sulfonyl group of (1).

In particular, the inventors of the present invention found that when said R is₂When the compound is halogen, the oxygen-containing substituted pyrazolopyrimidine compound with the structure shown in the formula (I) shows higher inhibitory activity on TRK kinase, particularly mutant TRK kinase; and simultaneously, the compound has more reasonable pharmacokinetic property and more excellent in-vivo anti-tumor activity. Therefore, the following embodiment 3, embodiment 4, embodiment 5 and embodiment 6 are provided to illustrate more excellent effects of the compound provided by the present invention.

Embodiment mode 3:

in the formula (I), the compound represented by the formula (I),

R₁、R₃and R₄Each independently selected from H, halogen, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl groups of (a);

R₂is halogen;

R₅selected from H, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, alkoxy-substituted C_2-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl groups of (a);

R₆is selected from C_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl and halogen of (a);

R₇is selected from C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl of, C_2-12Acyl group and sulfonyl group of (1).

Embodiment 4:

in the formula (I), the compound represented by the formula (I),

R₁、R₃and R₄Each independently selected from H, fluorine, chlorine, bromine, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl groups of (a);

R₂is F;

R₅selected from H, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl groups of (a);

R₆is selected from C_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl and halogen of (a);

R₇is selected from C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, cyano-substituted C_1-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl of, C_2-8Acyl group and sulfonyl group of (1).

Embodiment 5:

in the formula (I), the compound represented by the formula (I),

R₁、R₃and R₄Are all H; r₂Is F;

R₅selected from H, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl groups of (a);

R₆is selected from C_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl and halogen of (a);

R₇is selected from C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl of, C_2-8Acyl of (2)A sulfonyl group.

Embodiment 6:

the oxygen-containing substituted pyrazolopyrimidine compound is compound a1-2, compound a2-3, compound a3-4, compound a4-8, compound a5-9, compound a6-10, compound a7-12, compound a8-13, compound a9-14, compound a10-17, compound a11-18, compound a12-19, compound a13-20, compound a14-21, compound a15-22, compound a16-23, compound a17-24, compound a18-25, compound a19-26, compound a20-27, compound a21-28, compound a22-29, compound a23-31, compound a24-32, compound a25-35, compound a26-38, compound a27-41, compound a 3625-28, compound a22-29, compound a23-31, compound a24-32, compound a25-35, compound a26-38, At least one of compounds a28-43, or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, or prodrug thereof.

It is to be noted that, in the aforementioned compound a1-2, compound a2-3, compound a3-4, compound a4-8, compound a5-9, compound a6-10, compound a7-12, compound a8-13, compound a9-14, compound a10-17, compound a11-18, compound a12-19, compound a13-20, compound a14-21, compound a15-22, compound a16-23, compound a 16-24, compound a 16-25, compound a 16-26, compound a 16-27, compound a 16-28, compound a 16-29, compound a 16-31, compound a 16-32, compound a 16-35, compound a 16-38, compound a 16-41, and compound a 16-43, compounds not indicating a specific configuration of the chiral center, i.e. as racemates.

The method for producing the oxygen-containing substituted pyrazolopyrimidine compound having the structure represented by formula (I) is not particularly limited in the present invention, and for example, the following production method can be used:

the preparation method involves Suzuki coupling reaction, the reaction conditions of the coupling reaction are not particularly limited, and those skilled in the art can obtain appropriate reaction conditions according to common general knowledge in the field of organic synthesis and specific examples provided in the examples section of the present invention.

As described above, the second aspect of the present invention provides the use of the oxygen-containing substituted pyrazolopyrimidine compound having the structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof, described in the first aspect of the present invention, for the preparation of a medicament for the prevention and/or treatment of a TRK kinase receptor-mediated disease.

As described above, the third aspect of the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or diluent, and, as an active ingredient, an oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof, according to the first aspect of the present invention.

As mentioned above, a fourth aspect of the present invention provides the use of a pharmaceutical composition as described in the third aspect of the present invention in the manufacture of a medicament for the prevention and/or treatment of a TRK kinase receptor mediated disease.

As described above, the fifth aspect of the present invention provides an oxygen-containing substituted pyrazolopyrimidine compound having the structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof, according to the first aspect of the present invention, or a pharmaceutical composition according to the third aspect of the present invention, for use in the preparation of a medicament for the prevention and/or treatment of tumors.

Preferably, the tumor is at least one of breast cancer, large intestine cancer, lung cancer, thyroid cancer, skin cancer, bone cancer, melanoma, leukemia, salivary gland tumor, neuroendocrine tumor, lymphoma, brain tumor, neuroblastoma, ovarian cancer, pancreatic cancer, mesothelioma, esophageal cancer, pulmonary sarcoma, medulloblastoma, glioblastoma, colon cancer, liver cancer, retinoblastoma, kidney cancer, bladder cancer, osteosarcoma, stomach cancer, uterine cancer, vulval cancer, small intestine cancer, prostate cancer, bile duct cancer, ureteral cancer, adrenocortical cancer, or head and neck cancer.

The present invention will be described in detail below by way of examples. In the following examples, the various starting materials used are commercially available and are of analytical purity, unless otherwise specified.

Example 1: preparation of 4-fluoro-2- (1- ((3- (1-methyl-1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-yl) amino) ethyl) phenol (compound a1-2)

Step 1): 1- (5-fluoro-2-hydroxyphenyl) ethanone (32.4mmol), 4-methoxybenzyl bromide (38.9mmol), cesium carbonate (48.6mmol) were added to a 200mL pear-shaped vial, to which was added acetonitrile (50 mL). Heated overnight in an oil bath (80 ℃). The reaction was cooled to ambient temperature and the crude product was purified by column chromatography to give a white solid with 78% yield.

Step 2): 1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethanone (25.3mmol), hydroxylamine hydrochloride (30.4mmol), cesium carbonate (38.0mmol) were added to a 200mL pear-shaped bottle, to which methanol (50mL) was added. The reaction was carried out for 4h at room temperature with magnetic stirring. The reaction solution was poured into water, filtered with suction, and dried to obtain a white solid with a yield of 98%.

Step 3): 1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethanone oxime (24.8mmol), zinc powder (372mmol), ammonium chloride (372mmol), and acetic acid (372mmol) were charged to a 200mL pear-shaped bottle, to which methanol (50mL) was added. Heated overnight in an oil bath (80 ℃). The reaction was cooled to ambient temperature, neutralized, filtered through celite, and the filtrate was extracted with water and ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate and concentrated to give a yellow liquid with a yield of 77%.

Step 4): 5-Chloropyrazolo [1,5-a ] pyrimidine (19.1mmol), 1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethylamine (19.1mmol), anhydrous N-butanol (50mL) and N, N-diisopropylethylamine (DIPEA, 38.2mmol) were added to a 200mL pear-shaped vial. The mixture was heated overnight in an oil bath (140 ℃). The reaction was cooled to ambient temperature and concentrated under reduced pressure to remove N-butanol and N, N-Diisopropylethylamine (DIPEA) as much as possible to give a crude yellow oil which was purified by column chromatography to give a pale yellow solid with a yield of 68%.

Step 5): n- (1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (13.0mmol) was added to a 200mL pear-shaped vial, to which was added acetonitrile (50 mL). N-iodosuccinimide (NIS, 14.3mmol) was added under magnetic stirring at room temperature. The reaction was carried out at room temperature for 1h and TLC monitored for completion. After removing acetonitrile under reduced pressure as much as possible, the mixture was diluted with 250mL of ethyl acetate and transferred to a separatory funnel. Washing with 1mol/L NaOH for 3 times, washing with saturated salt for two times, drying with anhydrous sodium sulfate, concentrating to obtain red oily crude product, and purifying by column chromatography to obtain light yellow solid with yield of 67%.

Step 6): n- (1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethyl) -3-iodopyrazolo [1,5-a ] pyrimidin-5-amine (0.77mmol), 1-methylpyrazole-4-boronic acid pinacol ester (1.16mmol), anhydrous potassium carbonate (2.00mmol), tetrakis (triphenylphosphine) palladium (0.08mmol) were added to a 100ml reaction tube, replaced with argon 3 times, and 10ml anhydrous DMF,2ml water were added. The reaction was carried out at 100 ℃ for 2h under argon atmosphere and the completion of the reaction was monitored by TLC. Cooled to 50 ℃, filtered through celite, and the filtrate was extracted with water and ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, concentrated to give a crude product as a black oil, which was purified by column chromatography to give a pale yellow solid with a yield of 52%.

Step 7): n- (1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethyl) -3- (1-methyl-1H-pyrazol-3-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (0.22mmol) was added to a 200mL pear-shaped bottle, dichloromethane (10mL) was added thereto, and trifluoroacetic acid (3.3mmol) was added dropwise. The reaction was carried out for 4h at room temperature with magnetic stirring. Neutralized and extracted with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to give a white solid with a yield of 86%.

¹H NMR(400MHz,DMSO-d₆)δ9.68–9.65(m,1H),8.44(d,J＝7.6Hz,1H),8.02(s,1H),7.96(d,J＝6.8Hz,1H),7.84(s,1H),7.75(s,1H),7.04–6.77(m,3H),6.33(d,J＝7.6Hz,1H),5.44(d,J＝7.2Hz,1H),3.84(s,3H),1.43(d,J＝6.8Hz,3H).

Example 2: preparation of 4-fluoro-2- (1- ((3- (1- (2-hydroxycyclopentyl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-yl) amino) ethyl) phenol (compound a2-3)

Step 1): n- (1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethyl) -3- (1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (0.22mmol), 1, 2-epoxycyclopentane (0.22mmol), anhydrous DMF (10mL), and cesium carbonate (0.6mmol) were added to a solanaceous flask. The mixture was heated to reflux overnight in an oil bath (100 ℃). The reaction was cooled to ambient temperature, concentrated under reduced pressure to remove DMF as much as possible to give a yellow oily crude product, which was purified by column chromatography to give a pale yellow solid with a yield of 60%.

Step 2): the method of step 7 in example 1 was employed.

¹H NMR(400MHz,DMSO-d₆)δ9.60(d,J＝16.4Hz,1H),8.43(d,J＝7.6Hz,1H),8.03(d,J＝2.0Hz,1H),7.96–7.86(m,2H),7.82(s,1H),7.04–6.94(m,1H),6.88–6.77(m,2H),6.33(d,J＝7.6Hz,1H),5.54–5..44(m,1H),5.09–5..02(m,1H),4.42–4.14(m,2H),2.21–2.09(m,2H),2.02–1.87(m,2H),1.83–1.73(m,2H),1.63–1.54(m,2H),1.43(d,J＝6.8Hz,3H).

Example 3: preparation of 4-fluoro-2- (1- ((3- (1- (2-hydroxycyclohexyl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-yl) amino) ethyl) phenol (compound a3-4)

The procedure is as in example 2, except that 1, 2-epoxycyclopentane is replaced by 1, 2-epoxycyclohexane.

¹H NMR(400MHz,DMSO-d₆)δ9.59(d,J＝18.0Hz,1H),8.43(d,J＝7.6Hz,1H),8.01(d,J＝2.8Hz,1H),7.96–7.85(m,2H),7.80(s,1H),7.06–6.95(m,1H),6.82(dt,J＝6.4,1.6Hz,2H),6.42–6.22(m,1H),5.54–5.41(m,1H),4.84–4.64(m,1H),3.92–3.64(m,2H),2.03–1.65(m,6H),1.52–1.27(m,7H).

Example 4: preparation of (R) -N- (1- (2- (difluoromethoxy) -5-fluorophenyl) ethyl) -3- (1-methyl-1H-pyrazol-3-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a4-8)

Step 1): 5-Chloropyrazolo [1,5-a ] pyrimidine (19.1mmol), (R) -1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethylamine (19.1mmol), anhydrous N-butanol (50mL) and N, N-diisopropylethylamine (DIPEA, 38.2mmol) were added to a 200mL pear-shaped vial. The mixture was heated overnight in an oil bath (140 ℃). The reaction was cooled to ambient temperature and concentrated under reduced pressure to remove N-butanol and N, N-Diisopropylethylamine (DIPEA) as much as possible to give a crude yellow oil which was purified by column chromatography to give a pale yellow solid with a yield of 68%.

Step 2): (R) -N- (1- (5-fluoro-2- ((4-methoxybenzyl) oxy) phenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (13.0mmol) was added to a 200mL pear-shaped bottle, dichloromethane (50mL) was added thereto, and trifluoroacetic acid (195mmol) was added dropwise. The reaction was carried out for 4h at room temperature with magnetic stirring. Neutralized and extracted with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a white solid with a yield of 85%.

Step 3): (R) -4-fluoro-2- (1- (pyrazolo [1,5-a ] pyrimidin-5-ylamino) ethyl) phenol (11.0mmol), difluoromethane (16.5mmol), cesium carbonate (22mmol) were added to a 200mL pear-shaped bottle, to which DMF (50mL) was added. Heated overnight in an oil bath (100 ℃). The reaction was cooled to ambient temperature and the crude product was purified by column chromatography to give a white solid with 91% yield.

Step 4): (R) -N- (1- (5-fluoro-2-difluoromethoxy-phenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (9.8mmol) was added to a 200mL pear-shaped bottle, to which was added acetonitrile (50 mL). N-iodosuccinimide (NIS, 14.85mmol) was added under magnetic stirring at room temperature. The reaction was carried out at room temperature for 1h and TLC monitored for completion. After removing acetonitrile under reduced pressure as much as possible, the mixture was diluted with 250mL of ethyl acetate and transferred to a separatory funnel. Washing with 1mol/L NaOH for 3 times, washing with saturated salt for two times, drying with anhydrous sodium sulfate, concentrating to obtain red oily crude product, and purifying by column chromatography to obtain light yellow solid with yield of 67%.

Step 5): (R) -N- (1- (5-fluoro-2-difluoromethoxy-phenyl) ethyl) -3-iodopyrazolo [1,5-a ] pyrimidin-5-amine (0.50mmol), 1-methylpyrazole-4-boronic acid pinacol ester (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis (triphenylphosphine) palladium (0.05mmol) were added to a 100ml reaction tube, replaced with argon 3 times, and 10ml anhydrous DMF,2ml water were added. The reaction was carried out at 100 ℃ for 2h under argon atmosphere and the completion of the reaction was monitored by TLC. Cooled to 50 ℃, filtered through celite, and the filtrate was extracted with water and ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, concentrated to give a crude product as a black oil, which was purified by column chromatography to give a pale yellow solid with a yield of 52%.

¹H NMR(600MHz,DMSO-d₆)δ8.48(s,1H),8.07(d,J＝6.0Hz,2H),7.80(s,1H),7.71(s,2H),7.39–7.07(m,3H),6.34(s,1H),5.52–5.39(m,1H),3.83(s,3H),1.47d,J＝6.6Hz,3H).

Example 5: preparation of 2- (4- (5- ((1- (2- (difluoromethoxy) -5-fluorophenyl) ethyl) amino) pyrazolo [1,5-a ] pyrimidin-3-yl) -1H-pyrazolopiperazin-1-yl) ethan-1-ol (compound a5-9)

Step 1): 4-fluoro-2- (1- (pyrazolo [1,5-a ] pyrimidin-5-ylamino) ethyl) phenol (11.0mmol), difluoromethane (16.5mmol), cesium carbonate (22mmol) were added to a 200mL pear-shaped bottle, to which DMF (50mL) was added. Heated overnight in an oil bath (100 ℃). The reaction was cooled to ambient temperature and the crude product was purified by column chromatography to give a white solid with 91% yield.

Step 2): n- (1- (-5-fluoro-2-difluoromethoxyphenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (9.8mmol) was added to a 200mL pear-shaped bottle, to which was added acetonitrile (50 mL). N-iodosuccinimide (NIS, 14.85mmol) was added under magnetic stirring at room temperature. The reaction was carried out at room temperature for 1h and TLC monitored for completion. After removing acetonitrile under reduced pressure as much as possible, the mixture was diluted with 250mL of ethyl acetate and transferred to a separatory funnel. Washing with 1mol/L NaOH for 3 times, washing with saturated salt for two times, drying with anhydrous sodium sulfate, concentrating to obtain red oily crude product, and purifying by column chromatography to obtain light yellow solid with yield of 65%.

Step 3): n- (1- (5-fluoro-2-difluoromethoxyphenyl) ethyl) -3-iodopyrazolo [1,5-a ] pyrimidin-5-amine (0.50mmol), 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole-1-ethanol (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis (triphenylphosphine) palladium (0.05mmol) were added to a 100ml reaction tube, replaced with argon 3 times, and 10ml anhydrous DMF,2ml water were added. The reaction was carried out at 100 ℃ for 2h under argon atmosphere and the completion of the reaction was monitored by TLC. Cooled to 50 ℃, filtered through celite, and the filtrate was extracted with water and ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, concentrated to give a crude product as a black oil, which was purified by column chromatography to give a pale yellow solid with a yield of 52%.

¹H NMR(400MHz,DMSO-d6)δ8.50(d,J＝7.6Hz,1H),8.07(s,1H),8.06(s,1H),7.86(s,1H),7.76(s,1H),7.55–7.10(m,4H),6.37(d,J＝7.6Hz,1H),5.49(t,J＝7.2Hz,1H),4.95–4.88(m,1H),4.15(t,J＝6.0Hz,2H),3.77(q,J＝5.6Hz,2H),1.50(d,J＝7.2Hz,3H).

Example 6: preparation of N- (1- (2- (difluoromethoxy) -5-fluorophenyl) ethyl) -3- (1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a6-10)

A method similar to example 5 was used except that 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole-1-ethanol was replaced with 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester.

¹H NMR(400MHz,DMSO-d₆)δ8.50(d,J＝7.6Hz,1H),8.08(s,1H),8.06(s,1H),7.90(s,1H),7.78(s,1H),7.54–7.11(m,4H),6.38(d,J＝7.6Hz,1H),5.49(t,J＝7.0Hz,1H),4.36(dt,J＝10.0,4.4Hz,1H),4.03(dt,J＝11.2,3.2Hz,2H),3.50(dp,J＝11.2,4.4Hz,2H),1.98(td,J＝8.8,7.2,3.6Hz,4H),1.50(d,J＝6.8Hz,3H).

Example 7: preparation of N- (1- (2-ethoxy-5-fluorophenyl) ethyl) -3- (1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a7-12)

Step 1): 4-fluoro-2- (1- (pyrazolo [1,5-a ] pyrimidin-5-ylamino) ethyl) phenol (11.0mmol), iodoethane (16.5mmol), and cesium carbonate (22mmol) were added to a 200mL pear-shaped flask, to which DMF (50mL) was added. Heated overnight in an oil bath (100 ℃). The reaction was cooled to ambient temperature and the crude product was purified by column chromatography to give a white solid with 88% yield.

Step 2): n- (1- (5-fluoro-2-ethoxyphenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (9.8mmol) was added to a 200mL pear-shaped bottle, and acetonitrile (50mL) was added thereto. N-iodosuccinimide (NIS, 14.85mmol) was added under magnetic stirring at room temperature. The reaction was carried out at room temperature for 1h and TLC monitored for completion. After removing acetonitrile under reduced pressure as much as possible, the mixture was diluted with 250mL of ethyl acetate and transferred to a separatory funnel. Washing with 1mol/L NaOH solution for 3 times, washing with saturated salt solution for two times, drying with anhydrous sodium sulfate, concentrating to obtain red oily crude product, and purifying by column chromatography to obtain light yellow solid with yield of 68%.

Step 3): n- (1- (5-fluoro-2-ethoxyphenyl) ethyl) -3-iodopyrazolo [1,5-a ] pyrimidin-5-amine (0.50mmol), 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis (triphenylphosphine) palladium (0.05mmol) were added to a 100ml reaction tube, replaced with argon for 3 times, and 10ml anhydrous DMF,2ml water were added. The reaction was carried out at 100 ℃ for 2h under argon atmosphere and the completion of the reaction was monitored by TLC. Cooled to 50 ℃, filtered through celite, and the filtrate was extracted with water and ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, concentrated to give a crude product as a black oil, which was purified by column chromatography to give a pale yellow solid with a yield of 52%.

¹H NMR(400MHz,DMSO-d6)δ8.48(d,J＝7.2Hz,1H),8.06(s,1H),7.98(d,J＝7.0Hz,1H),7.87(s,1H),7.82(s,1H),7.15–6.92(m,3H),6.38(d,J＝7.2Hz,1H),5.61–5.44(m,1H),4.44–3.92(m,5H),3.59–3.43(m,2H),1.96(s,4H),1.45(d,J＝7.2Hz,6H).

Example 8: preparation of 2- (2- (1- ((3- (1- (difluoromethyl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-yl) amino) ethyl) -4-fluorophenoxy) ethanol (compound a8-13)

A method similar to example 7 was used, except that iodoethane was replaced with iodoethanol, and that 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester was replaced with 1- (difluoromethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) -1H-pyrazole.

¹H NMR(400MHz,DMSO-d₆)δ8.52(d,J＝7.6Hz,1H),8.29(s,1H),8.19(s,1H),8.16(s,1H),8.09(d,J＝7.2Hz,1H),7.82(t,J＝59.6Hz,1H),7.12–6.91(m,3H),6.41(d,J＝7.6Hz,1H),5.59–5.46(m,1H),5.00(t,J＝5.6Hz,1H),4.23–4.03(m,2H),3.90–3.80(m,2H),1.48(d,J＝6.8Hz,3H).

Example 9: preparation of N- (1- (5-fluoro-2- (2-methoxyethoxy) phenyl) ethyl) -3- (1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a9-14)

A method similar to example 7 was used except that iodoethane was replaced with 2-iodoethyl methyl ether.

¹H NMR(400MHz,DMSO-d₆)δ8.45(d,J＝7.6Hz,1H),8.03(s,1H),7.90(d,J＝7.2Hz,1H),7.82(s,1H),7.77(s,1H),7.08–6.94(m,3H),6.35(d,J＝7.6Hz,1H),5.54–5.43(m,1H),4.39–4.13(m,3H),4.03–3.94(m,2H),3.81–3.69(m,2H),3.53–3.44(m,2H),3.33(s,3H),1.99–1.83(m,4H),1.45(d,J＝6.8Hz,3H).

Example 10: preparation of (R) -N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) -3- (1-methyl-1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a10-17)

A similar procedure to that of example 4 was followed, except that difluoromethane was replaced with 1-fluoro-2-iodoethane.

¹H NMR(400MHz,DMSO-d₆)δ8.49(d,J＝7.2Hz,1H),8.05(s,1H),8.01(d,J＝7.2Hz,1H),7.76(s,2H),7.17–6.95(m,3H),6.38(d,J＝7.6Hz,1H),5.61–5.49(m,1H),4.99–4.90(m,1H),4.87–4.78(m,1H),4.51–4.32(m,2H),3.84(s,3H),1.47(d,J＝6.8Hz,3H).

Example 11: preparation of 3- (1- (difluoromethyl) -1H-pyrazol-4-yl) -N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a11-18)

A method similar to example 7 was used, except that iodoethane was replaced with 1-fluoro-2-iodoethane, and that 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester was replaced with 1- (difluoromethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole.

¹H NMR(400MHz,DMSO-d₆)δ8.53(d,J＝7.2Hz,1H),8.26(s,1H),8.19(s,1H),8.12(d,J＝8.0Hz,2H),7.80(t,J＝59.2Hz,1H),7.05(m,3H),6.43(d,J＝7.6Hz,1H),5.58–5.49(m,1H),4.96–4.85(m,1H),4.83–4.74(m,1H),4.48–4.37(m,1H),4.38–4.30(m,1H),1.48(d,J＝6.8Hz,3H).

Example 12: preparation of (R) -2- (4- (5- ((1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) amino) pyrazolo [1,5-a ] pyrimidin-3-yl) -1H-pyrazol-1-yl) acetonitrile (compound a12-19)

A method similar to example 4 was used, except that difluoromethane was replaced with 1-fluoro-2-iodoethane and 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester was replaced with 2- [4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl ] acetonitrile, respectively.

¹H NMR(400MHz,DMSO-d₆)δ8.48(d,J＝7.2Hz,1H),8.08(s,1H),8.04(s,1H),7.91(d,J＝8.0Hz,2H),7.20–6.93(m,3H),6.37(d,J＝7.2Hz,1H),5.59–5.50(m,1H),5.47–5.40(m,2H),4.96–4.87(m,1H),4.84–4.74(m,1H),4.48–4.30(m,2H),1.45(d,J＝6.8Hz,3H).

Example 13: preparation of N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) -3- (1-isopropyl-1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a13-20)

The procedure is as in example 11, except that 1- (difluoromethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole is replaced with 1-isopropyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole.

1H NMR(400MHz,DMSO-d6)δ8.45(d,J＝7.6Hz,1H),8.01(s,1H),7.91(d,J＝7.2Hz,1H),7.81(s,1H),7.73(s,1H),7.13–6.94(m,3H),6.35(d,J＝7.6Hz,1H),5.55–5.42(m,1H),4.89(t,J＝4.0Hz,1H),4.77(t,J＝4.0Hz,1H),4.49–4.28(m,3H),1.45(d,J＝6.8Hz,3H),1.40(d,J＝6.8Hz,6H).

Example 14: preparation of 3- (1-cyclopropyl-1H-pyrazol-4-yl) -N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a14-21)

The procedure is as in example 11, except that 1- (difluoromethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole is replaced with 1-cyclopropylpyrazole-4-boronic acid pinacol ester.

¹H NMR(400MHz,DMSO-d₆)δ8.48(d,J＝7.6Hz,1H),8.04(s,1H),7.99(d,J＝6.8Hz,1H),7.81(s,1H),7.74(s,1H),7.17–6.97(m,3H),6.38(d,J＝7.6Hz,1H),5.57–5.46(m,1H),4.97–4.89(m,1H),4.84–4.78(m,1H),4.51–4.31(m,2H),3.71–3.60(m,1H),1.47(d,J＝6.8Hz,3H),1.06–0.95(m,4H).

Example 15: preparation of 2- (4- (5- ((1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) amino) pyrazolo [1,5-a ] pyrimidin-3-yl) -1H-pyrazol-1-yl) ethanol (compound a15-22)

The procedure is as in example 11, except that 1- (difluoromethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole is replaced with 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole-1-ethanol.

¹H NMR(400MHz,DMSO-d₆)δ8.48(d,J＝7.6Hz,1H),8.05(s,1H),7.98(d,J＝7.2Hz,1H),7.80(s,1H),7.77(s,1H),7.11–7.00(m,3H),6.37(d,J＝7.6Hz,1H),5.60–5.51(m,1H),4.96–4.91(m,1H),4.84–4.79(m,1H),4.54–4.31(m,2H),4.11(t,J＝5.6Hz,2H),3.73(t,J＝5.6Hz,2H),1.47(d,J＝6.8Hz,3H).

Example 16: preparation of 3- (1- (2, 2-difluoroethyl) -1H-pyrazol-4-yl) -N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a16-23)

The procedure is as in example 11, except that 1- (difluoromethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole is replaced by 1- (2, 2-difluoroethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole.

¹H NMR(400MHz,DMSO-d₆)δ8.49(d,J＝7.6Hz,1H),8.08(s,1H),8.01(d,J＝7.2Hz,1H),7.89(s,1H),7.87(s,1H),7.19–6.96(m,3H),6.55–6.19(m,2H),5.60–5.50(m,1H),4.92(t,J＝4.0Hz,1H),4.80(t,J＝3.6Hz,1H),4.67–4.52(m,2H),4.42(d,J＝4.8Hz,1H),4.35(d,J＝4.8Hz,1H),1.47(d,J＝6.8Hz,3H).

Example 17: preparation of 2- (4- (5- ((1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) amino) pyrazolo [1,5-a ] pyrimidin-3-yl) -1H-pyrazol-1-yl) cyclopentan-1-ol (compound a17-24)

N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) -3- (1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (0.22mmol), 1, 2-epoxycyclopentane (0.22mmol), anhydrous DMF (10mL), and cesium carbonate (0.6mmol) were added to a solanaceous bottle. The mixture was heated to reflux overnight in an oil bath (100 ℃). The reaction was cooled to ambient temperature, concentrated under reduced pressure to remove DMF as much as possible to give a yellow oily crude product, which was purified by column chromatography to give a pale yellow solid with a yield of 65%.

¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝7.6Hz,1H),8.04(s,1H),7.97(s,1H),7.83(d,J＝5.6Hz,1H),7.79(s,1H),7.15–6.97(m,3H),6.37(d,J＝7.6Hz,1H),5.58–5.49(m,1H),5.07(t,J＝5.6Hz,1H),4.97–4.89(m,1H),4.84–4.77(m,1H),4.51–4.15(m,4H),2.21–2.10(m,1H),2.02–1.90(m,2H),1.85–1.73(m,2H),1.64–1.54(m,1H),1.47(d,J＝6.8Hz,3H).

Example 18: preparation of 2- (4- (5- ((1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) amino) pyrazolo [1,5-a ] pyrimidin-3-yl) -1H-pyrazol-1-yl) cyclohexan-1-ol (compound a18-25)

The procedure is as in example 17, except that 1, 2-epoxycyclopentane is replaced by 1, 2-epoxycyclohexane.

¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝7.6Hz,1H),8.03(s,1H),7.95(d,J＝7.2Hz,1H),7.79(s,1H),7.75(d,J＝2.4Hz,1H),7.16–6.97(m,3H),6.37(d,J＝7.6Hz,1H),5.59–5.48(m,1H),4.98–4.69(m,3H),4.51–4.31(m,2H),3.86–3.66(m,2H),2.07–1.67(m,5H),1.47(d,J＝7.2Hz,3H),1.40–1.24(m,3H).

Example 19: preparation of N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) -3- (1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a19-26)

The preparation is as in example 11, except that 1- (difluoromethyl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole is replaced by 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester.

¹H NMR(400MHz,DMSO-d₆)δ8.46(d,J＝7.8Hz,1H),8.04(s,1H),7.94(d,J＝7.2Hz,1H),7.83(s,1H),7.78(s,1H),7.16–6.94(m,3H),6.38(d,J＝7.6Hz,1H),5.59–5.41(m,1H),4.91(dt,J＝4.8,2.4Hz,1H),4.79(dt,J＝4.4,2.4Hz,1H),4.51–4.25(m,3H),4.00(dq,J＝10.8,3.2Hz,2H),3.49(dp,J＝10.8,3.6Hz,2H),1.99–1.81(m,4H),1.47(d,J＝6.8Hz,3H).

Example 20: preparation of N- ((R) -1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a20-27)

The preparation method was the same as in example 10, except that 1-methylpyrazole-4-boronic acid pinacol ester was replaced with 1- (tetrahydropyran-2-yl) -4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) -1H-pyrazole.

¹H NMR(400MHz,DMSO-d₆)δ8.49(d,J＝7.6Hz,1H),8.09(s,1H),8.00(d,J＝6.8Hz,1H),7.96(s,1H),7.85(s,1H),7.15–6.96(m,3H),6.38(d,J＝7.6Hz,1H),5.57–5.47(m,1H),5.39–5.32(m,1H),4.96–4.91(m,1H),4.84–4.78(m,1H),4.46–4.41(m,1H),4.39–4.34(m,1H),3.99–3.91(m,1H),3.71–3.62(m,1H),2.16–1.88(m,3H),1.78–1.68(m,1H),1.61–1.54(m,2H),1.47(d,J＝6.8Hz,3H).

Example 21: preparation of 1- (4- (5- ((1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) amino) pyrazolo [1,5-a ] pyrimidin-3-yl) -1H-pyrazol-1-yl) ethanone (compound a21-28)

To a solanaceous bottle were added N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) -3- (1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (0.26mmol), acetyl chloride (0.26mmol), anhydrous DCM (10mL) and triethylamine (0.52 mmol). The reaction was carried out at 0 ℃ for 4 h. Vacuum concentrating to obtain crude product, and purifying by column chromatography to obtain light yellow solid with yield of 70%.

¹H NMR(400MHz,DMSO-d₆)δ8.54(d,J＝7.6Hz,1H),8.47(s,1H),8.29(s,1H),8.27(s,1H),8.16(d,J＝7.2Hz,1H),7.15–6.96(m,3H),6.43(d,J＝7.6Hz,1H),5.62–5.53(m,1H),4.99–4.90(m,1H),4.87–4.78(m,1H),4.57–4.32(m,2H),2.66(s,3H),1.47(d,J＝6.8Hz,3H).

Example 22: preparation of N- (1- (5-fluoro-2- (2-fluoroethoxy) phenyl) ethyl) -3- (1- (methylsulfonyl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a22-29)

The procedure is as in example 21, except that acetyl chloride is replaced by methanesulfonyl chloride.

¹H NMR(400MHz,DMSO-d₆)δ8.54(d,J＝7.6Hz,1H),8.37(s,1H),8.32(s,1H),8.26(s,1H),8.15(d,J＝7.2Hz,1H),7.11–6.99(m,3H),6.44(d,J＝7.6Hz,1H),5.61–5.51(m,1H),4.97–4.89(m,1H),4.86–4.75(m,1H),4.53–4.29(m,2H),3.51(s,3H),1.48(d,J＝6.8Hz,3H).。

Example 23: preparation of (R) -N- (1- (5-fluoro-2- (2,2, 2-trifluoroethoxy) phenyl) ethyl) -3- (1-methyl-1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a23-31)

A similar procedure to that of example 4 was followed, except that difluoromethane was replaced with 2-iodo-1, 1, 1-trifluoroethane.

¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝7.6Hz,1H),8.02(s,1H),8.00(d,J＝6.8Hz,1H),7.70(d,J＝2.4Hz,2H),7.25–7.01(m,3H),6.35(d,J＝7.6Hz,1H),5.53–5.41(m,1H),4.99–4.81(m,2H),3.80(s,3H),1.44(d,J＝6.4Hz,3H).

Example 24: preparation of (R) -2- (4- (5- ((1- (5-fluoro-2- (2,2, 2-trifluoroethoxy) phenyl) ethyl) amino) pyrazolo [1,5-a ] pyrimidin-3-yl) -1H-pyrazol-1-yl) ethanol (compound a24-32)

A method similar to example 4 was used, except that iodoethane was replaced with 2-iodo-1, 1, 1-trifluoroethane, and that 1-methylpyrazole-4-boronic acid pinacol ester was replaced with 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole-1-ethanol.

¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝7.2Hz,1H),8.03(s,1H),7.98(d,J＝6.4Hz,1H),7.73(d,J＝11.2Hz,2H),7.27–7.00(m,3H),6.35(d,J＝7.6Hz,1H),5.51–5.41(m,1H),5.00–4.78(m,3H),4.08(t,J＝5.6Hz,2H),3.71(q,J＝5.6Hz,2H),1.44(d,J＝6.8Hz,3H).

Example 25: preparation of N- (1- (5-fluoro-2- (3-fluoropropoxy) phenyl) ethyl) -3- (1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a25-35)

A similar procedure to example 7 was used, except that iodoethane was replaced with 3-fluoro-1-iodopropane.

1H NMR(400MHz,DMSO-d6)δ8.46(d,J＝7.6Hz,1H),8.03(s,1H),7.93(d,J＝7.2Hz,1H),7.84(s,1H),7.79(s,1H),7.12–6.94(m,3H),6.36(d,J＝7.6Hz,1H),5.49(t,J＝7.2Hz,1H),4.73(td,J＝6.0,2.4Hz,1H),4.61(td,J＝6.0,2.4Hz,1H),4.40–4.07(m,3H),3.99(dq,J＝11.2,3.2Hz,2H),3.49(tt,J＝11.2,3.2Hz,2H),2.22(dp,J＝24.4,6.0Hz,2H),1.92(td,J＝11.2,10.4,4.4Hz,4H),1.45(d,J＝6.8Hz,3H).

Example 26: preparation of N- (1- (2- (cyclopropylmethoxy) -5-fluorophenyl) ethyl) -3- (1-methyl-1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a26-38)

A method similar to example 7 was used, except that iodoethane was replaced with iodomethylcyclopropane, and that 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester was replaced with 1-methylpyrazole-4-boronic acid pinacol ester.

¹H NMR(400MHz,DMSO-d₆)δ8.48(d,J＝7.6Hz,1H),8.03(s,1H),7.99(d,J＝6.8Hz,1H),7.76(d,J＝2.8Hz,2H),7.14–6.90(m,3H),6.37(d,J＝7.6Hz,1H),5.58–5.49(m,1H),4.09–3.93(m,2H),3.84(s,3H),1.47(d,J＝6.8Hz,3H),1.39–1.29(m,1H),0.64–0.56(m,2H),0.46–0.39(m,2H).

Example 27: preparation of N- (1- (5-fluoro-2- ((tetrahydrofuran-3-yl) oxy) phenyl) ethyl) -3- (1-methyl-1H-pyrazol-4-yl) pyrazolo [1,5-a ] pyrimidin-5-amine (compound a27-41)

A method similar to example 7 was used, except that iodoethane was replaced with 3-iodotetrahydrofuran, and that 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester was replaced with 1-methylpyrazole-4-boronic acid pinacol ester.

¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝7.6Hz,1H),8.03(s,1H),7.94(d,J＝7.0Hz,1H),7.78(s,1H),7.75(s,1H),7.16–6.94(m,3H),6.37(d,J＝7.6Hz,1H),5.51–5.43(m,1H),5.21–5.15(m,1H),4.10–3.76(m,7H),2.40–1.96(m,2H),1.44(d,J＝6.8Hz,3H).

Example 28: preparation of N- (1- (5-fluoro-2- ((tetrahydro-2H-pyran-4-yl) methoxy) phenyl) ethyl) -3- (1-methyl-1H-pyrazol-4-yl) pyrazolo [1,25-a ] pyrimidin-5-amine (compound a28-43)

A method similar to example 7 was used, except that iodoethane was replaced with 4-iodomethyltetrahydropyran, and that 1- (tetrahydropyran-4-yl) -1H-pyrazole-4-boronic acid pinacol ester was replaced with 1-methylpyrazole-4-boronic acid pinacol ester.

¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝7.6Hz,1H),8.02(s,1H),7.95(d,J＝6.8Hz,1H),7.75(s,2H),7.14–6.93(m,3H),6.36(d,J＝7.6Hz,1H),5.58–5.48(m,1H),4.07–3.98(m,1H),3.97–3.77(m,6H),3.31–3.28(m,2H),2.18–2.06(m,1H),1.83–1.67(m,2H),1.55–1.35(m,5H).

Test example 1: in vitro biochemical level inhibition Protein Kinase (PK) activity experiment

The material and the method are as follows: wild type kinases such as TRKA, TRKB and TRKC, and mutant kinases such as TRKA-G595R, TRKA-G667C, TRKA-F589L, TRKC-G623R and TRKC-G696A, which are derived from Carna Biosciences 08-186, 08-187, 08-197; HTRF KinEASE TKkit (Cisbio 62TK0 PEC); 384 well plates (Greiner corporation); ATP (Life technologies PV3227), MgCl₂(sigma) corporation; PHERAstar FS multifunctional microplate reader (BMG company); low speed centrifuge (StaiteXiangyi corporation); incubator (Binder Co.).

The control compounds selected were typical compound B disclosed in typical compound A, WO2007025540 disclosed in WO2007147647, and comparative compound C, comparative compound D, wherein comparative compound C (nuclear magnetic data is:¹H NMR(400MHz,DMSO-d₆) δ 12.78-12.66 (m,1H),8.45(D, J ═ 7.6Hz,1H),8.05(s,1H), 8.01-7.80 (m,3H),6.94(s,1H),6.90(D, J ═ 9.2Hz,1H),6.77(dt, J ═ 10.8,2.4Hz,1H), 6.56-6.21 (m,2H),5.20(D, J ═ 6.0Hz,1H),4.31(tt, J ═ 14.4,3.6Hz,2H),1.48(D, J ═ 6.8Hz,3H)) and comparative compound D (data:¹H NMR(400MHz,DMSO-d₆) δ 12.70(br,1H),8.44(d, J ═ 7.6Hz,1H),8.05(s,1H),7.94(d, J ═ 6.8Hz,2H),7.85(s,1H),7.35 to 7.14(m,3H),6.54 to 6.20(m,2H),5.28 to 5.11(m,1H),4.39 to 4.30(m,2H),1.48(d, J ═ 6.8Hz,3H)) the procedure used for the preparation of example 1 is as follows, except that the starting materials are different:

compound dissolution and preservation: preparing a test compound into a mother solution of 10mmol/L by using dimethyl sulfoxide (DMSO) according to the solubility, subpackaging and storing at-20 ℃;

preparing a compound working solution: before testing, the dispensed compound was removed from the freezer and diluted to 100 × the desired concentration with pure DMSO; then the compound was diluted to 4 x the desired concentration with deionized water;

1.33 Xpreparation of enzyme buffer (enzymic buffer): the 5 x enzyme buffer (from HTRF kit) was diluted 1.33 x with deionized water and 1.33 x final concentration of the corresponding ingredients was added: 1.33mmol/L Dithiothreitol (DTT) and 1.33mmol/L MnCl₂、6.65mmol/L MgCl₂And 39.9nmol/L SEB;

preparation of a kinase working solution: TRKA, TRKB and TRKC were diluted to 2 Xthe desired concentrations of 0.404 ng/. mu.L, 0.304 ng/. mu.L and 0.236 ng/. mu.L with 1.33 Xenzyme buffer;

preparing a substrate working solution: TK Substrate-biotin (from HTRF kit) and ATP (10mM) were diluted to 4X the desired final concentration in 1.33 Xenzyme buffer; the final ATP concentrations for TRKA, TRKB, and TRKC are: 3.727. mu. mol/L, 2.56. mu. mol/L and 2.526. mu. mol/L. TK Substrate-biotin (from HTRF KinEASE TKkit) final concentrations were: 0.2. mu. mol/L.

Preparation of detection working solution: 16.67. mu. mol/L of Streptavidin-XL665 (Streptavidin-XL665) were diluted to 4 Xthe desired final concentration with HTRF test buffer and then mixed with an equal volume of Antibody europium Cryptate (Antibody-Cryptate) (both from HTRF kits).

An enzyme reaction step: add 4. mu.L of kinase working solution to each well of a low volume 384 microwell plate, while adding 4. mu.L of 1.33 Xenzyme buffer as a Negative control (Negative); add 2. mu.l of compound working solution to the wells, while adding 2. mu.l of 8% DMSO aqueous solution as a zero compound concentration control (i.e., Positive control); incubating at 25 deg.C for 5 min; add 2. mu.L of substrate working solution to the wells to start the enzymatic reaction, shake the reaction for 30min at 37 ℃.

HTRF reagent detection step: adding 8 mu L of detection working solution into the hole to terminate the reaction; reacting for 1h at 25 ℃;

reading of HTRF signal: the PHERAStar FS reading is adopted to detect signals, and the corresponding settings of the instrument are as follows:

Optic module

integration delay (lag time)50 μ s

Integration time (Integration time) 400. mu.s

Flash Number of flash (Number of flashes)200

For the raw data read out per well, the ratio is 665nm/620 nm;

calculation of inhibition ratio:

IC₅₀calculation of the value: a non-linear curve was fitted in GraphPad Prism5 with the logarithm of the compound concentration as abscissa and the inhibition as ordinate: log (inhibitor) vs. response-Variable slope, and determining the concentration of the compound to be tested, namely IC when the enzyme activity inhibition rate is 50 percent₅₀。

The experimental results are as follows: TRKA, TRKB and TRKC kinase activity half Inhibitory Concentration (IC)₅₀,nM)

The invention provides the half Inhibitory Concentrations (IC) of compounds having the structure shown in formula (I) and control compounds on TRKA, TRKB and TRKC₅₀) See table 1:

table 1: TRKA, TRKB and TRKC kinase inhibitory Activity of Compounds

As shown in table 1, the compounds provided by the present invention all exhibit excellent inhibitory activity against wild TRKA, TRKB and TRKC kinases, which is significantly superior to typical compound a, typical compound B, comparative compound C and comparative compound D.

Test example 2: drug metabolism study in rats

The compounds provided in the previous section of the examples were administered to rats as polyethylene glycol 400 in water (70%). For oral administration, rats were given a dose of 5 mg/kg. Approximately 0.3mL of each blood sample was collected 15, 30, 45min, 1,2, 4,6, 8, 10, 24h after oral group administration into heparinized Eppendorf tubes, buffered on ice and centrifuged. The whole blood was centrifuged at 8000rpm for 5min and plasma was collected, transferred to a 96 well plate and stored at-20 ℃ until detection by LC-MS/MS.

The pharmacokinetic parameters after administration in rats were calculated using a non-compartmental model of the software WinNonlin software.

Peak concentration Cmax: adopting an actual measurement value;

AUC0-t value of area under the curve at time of drug: calculating by adopting a trapezoidal method; AUC_0-∞＝AUC_0-t+ Ct/ke, where Ct is the blood concentration at the last measurable time point and ke is the elimination rate constant;

elimination of half-life t_1/2＝0.693/ke；

Absolute bioavailability of F ═ Dose_iv*AUC_0-t，ig/Dose_ig*AUC_0-t，iv×100％。

Table 2 lists the pharmacokinetic parameters of the compounds of the invention in rats after intravenous administration. The results indicate that the compounds of the invention have good pharmacokinetic properties, including ideal Clearance (CL), half-life (t)_1/2) Peak concentration (C)_max) And exposure (AUC)_0-t)。

Table 3 lists the pharmacokinetic parameters of the compounds of the invention in rats after oral administration. The results indicate that the compounds of the invention have good pharmacokinetic properties, including ideal Clearance (CL), half-life (t)_1/2) Peak concentration (C)_max) Exposure (AUC)_0-t) And oral bioavailability.

Table 2: primary pharmacokinetic parameters for rat intravenous administration of Compound a24-32

Table 3: primary pharmacokinetic parameters for oral rat administration of Compound a24-32

Test example 3: inhibitory Activity of Compounds of the present invention against five TRK kinase mutants

This test example was conducted by the same test method as in test example 1.

The results of this test example are shown in Table 4.

TABLE 4

As can be seen from Table 4, the inhibitory activity of the compound of the invention on five TRK kinase mutants is superior to that on wild TRK kinase, and the compound is expected to effectively overcome the tumor drug resistance reported in clinic.

The above results show that the oxygen-containing substituted pyrazolopyrimidine compound having the structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, or a prodrug thereof, shows excellent inhibitory activity on TRK kinase, and at the same time, can show good antitumor activity at an animal level.

The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.

Claims (13)

1. An oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

wherein, in the formula (I),

R₁、R₃and R₄Each independently selected from H, halogen, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl groups of (a);

R₂is F;

R₅selected from H, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, alkoxy-substituted C_2-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl groups of (a);

R₆selected from H, C_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl and halogen of (a);

R₇is selected from C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl of, C_2-12Acyl, sulfonyl of (a);

wherein the content of the first and second substances,

said alkoxy-substituted C_2-12Represents a group having 2 to 12 carbon atoms in total, and the alkoxy group is substituted C_2-12The alkyl group of (A) has the formula-R¹OR²Wherein, said R¹And said R²The sum of the number of carbon atoms in (A) is 2 to 12, and the R¹Directly connecting with the phenoxy in the oxygen-containing substituted pyrazolopyrimidine compound with the structure shown in the formula (I);

said cyano-substituted C_2-12Alkyl of (2) represents an alkane having 2 to 12 carbon atoms in totalGroup, said cyano-substituted C_2-12Is a straight-chain alkyl group, a branched-chain alkyl group or a cyclic alkyl group, and the cyano group is substituted C_2-12At least one H in the alkyl group of (a) is substituted by a cyano group, and the carbon atom in the cyano group is counted as C substituted by the cyano group_2-12The total number of carbon atoms in the alkyl group of (a);

c containing 1-3 heteroatoms selected from N, O and S_2-12Represents cycloalkyl having a total number of carbon atoms of 2 to 12 and forms said C containing 1 to 3 heteroatoms selected from N, O and S_2-121-3 of the heteroatoms of cycloalkyl of (a) are selected from the group consisting of N, O and S, and form the C containing 1-3 heteroatoms selected from the group consisting of N, O and S_2-12Optionally contains an alkyl substituent on the atom, the number of carbon atoms contained in said alkyl substituent counting from 1 to 3 of said C containing a heteroatom selected from N, O and S_2-12In the total number of carbon atoms of said cycloalkyl group, and said C containing 1 to 3 hetero atoms selected from N, O and S_2-12H in the cycloalkyl group of (a) is optionally substituted or unsubstituted with a substituent, and if substituted, each of the substituents is independently selected from at least one of halogen, hydroxyl, nitro and mercapto.

2. The compound according to claim 1, wherein, in formula (I),

R₁、R₃and R₄Each independently selected from H, fluorine, chlorine, bromine, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl groups of (a);

R₂is F;

R₅selected from H, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl groups of (a);

R₆selected from H, C_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl and halogen ofA peptide;

R₇is selected from C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl of, C_2-8Acyl group and sulfonyl group of (1).

3. The compound according to claim 2, wherein, in formula (I),

R₁、R₃and R₄Each independently selected from H, fluorine, chlorine, bromine, C_1-6Alkyl of (2), C substituted by 1 to 4 halogens selected from fluorine, chlorine and bromine_1-6Alkyl groups of (a);

R₂is F;

R₅selected from H, C_1-6Alkyl of (2), C substituted by 1 to 4 halogens selected from fluorine, chlorine and bromine_1-6Alkyl, hydroxy-substituted C of_1-6Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-6Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-8Cycloalkyl groups of (a);

R₆selected from H, C_1-6Alkyl, hydroxy-substituted C of_1-6Alkyl and halogen of (a);

R₇is selected from C_1-6Alkyl of (2), C substituted by 1 to 4 halogens selected from fluorine, chlorine and bromine_1-6Alkyl, hydroxy-substituted C of_1-6Alkyl, cyano-substituted C_2-6Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-8Cycloalkyl of, C_2-6Acyl group and sulfonyl group of (1).

4. The compound according to claim 1, wherein, in formula (I),

R₁、R₃and R₄Each independently selected from H, halogen, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl groups of (a);

R₂is F;

R₅selected from H, C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, alkoxy-substituted C_2-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl groups of (a);

R₆is selected from C_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl and halogen of (a);

R₇is selected from C_1-12Alkyl of (2), C substituted by 1-6 halogens_1-12Alkyl, hydroxy-substituted C of_1-12Alkyl, cyano-substituted C_2-12Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-12Cycloalkyl of, C_2-12Acyl group and sulfonyl group of (1).

5. The compound according to claim 4, wherein, in formula (I),

R₁、R₃and R₄Each independently selected from H, fluorine, chlorine, bromine, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl groups of (a);

R₂is F;

R₅selected from H, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl groups of (a);

R₆is selected from C_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl and halogen of (a);

R₇is selected from C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, cyano-substituted C_1-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl of, C_2-8Acyl group of (2)And a sulfonyl group.

6. The compound according to claim 4, wherein, in formula (I),

R₁、R₃and R₄Are all H; r₂Is F;

R₅selected from H, C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, alkoxy-substituted C_2-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl groups of (a);

R₆is selected from C_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl and halogen of (a);

R₇is selected from C_1-8Alkyl of (2), C substituted by 1-6 halogens selected from fluorine, chlorine and bromine_1-8Alkyl, hydroxy-substituted C of_1-8Alkyl, cyano-substituted C_2-8Alkyl of (a), C containing 1-3 heteroatoms selected from N, O and S_2-10Cycloalkyl of, C_2-8Acyl group and sulfonyl group of (1).

7. The compound of claim 1, wherein the oxygen-containing substituted pyrazolopyrimidine compound is at least one of the following compounds, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

compound a 1-2:

compound a 2-3:

compound a 3-4:

compound a 4-8:

compound a 5-9:

compound a 6-10:

compound a 7-12:

compound a 8-13:

compound a 9-14:

compound a 10-17:

compound a 11-18:

compound a 12-19:

compound a 13-20:

compound a 14-21:

compound a 15-22:

compound a 16-23:

compound a 17-24:

compound a 18-25:

compound a 19-26:

compound a 20-27:

compound a 21-28:

compound a 22-29:

compound a 23-31:

compound a 24-32:

compound a 25-35:

compound a 26-38:

compound a 27-41:

compound a 28-43:

8. use of an oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 7 in the preparation of a medicament for the prevention and/or treatment of a TRK kinase-mediated disease.

9. A pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or diluent, and as an active ingredient an oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 7.

10. Use of the pharmaceutical composition according to claim 9 for the preparation of a medicament for the prevention and/or treatment of a TRK kinase receptor mediated disease.

11. Use of an oxygen-containing substituted pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 7, or a pharmaceutical composition according to claim 9 for the preparation of a medicament for the prevention and/or treatment of tumors.

12. The use of claim 11, wherein the tumor is at least one of breast cancer, large intestine cancer, lung cancer, thyroid cancer, skin cancer, bone cancer, melanoma, leukemia, salivary gland tumor, neuroendocrine tumor, lymphoma, brain tumor, ovarian cancer, pancreatic cancer, mesothelioma, esophageal cancer, lung sarcoma, medulloblastoma, glioblastoma, colon cancer, liver cancer, retinoblastoma, kidney cancer, bladder cancer, stomach cancer, uterine cancer, vulval cancer, small intestine cancer, prostate cancer, bile duct cancer, ureter cancer, adrenocortical cancer, or head and neck cancer.

13. The use of claim 11, wherein the tumor is at least one of neuroblastoma, osteosarcoma.