Macrocyclic compound, pharmaceutical composition and use thereof

Abstract

The present invention provides a compound as shown in formula (I) or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof. The present invention also provides a pharmaceutical composition comprising the compound and the use of the compound and the pharmaceutical composition in the preparation of a drug for treating a tyrosine kinase-mediated disease. The compound and the pharmaceutical composition provided by the present invention have significant tyrosine kinase inhibitory activity, can overcome tumor drug resistance, can break through the blood-brain barrier, and also have excellent pharmacokinetic properties and excellent oral bioavailability. They can be administered in smaller doses, thereby reducing the patient's treatment costs and possible toxic side effects, and thus have great application potential.

Description

Technical Field

[0001] The present invention relates to certain macrocyclic compounds that inhibit multiple kinases such as SRC, MET and / or CSF1R; pharmaceutical compositions containing such compounds; and methods for treating cancer using such compounds. Background Art

[0002] Protein kinases are key regulators of cell growth, proliferation and survival. Genetic and epigenetic changes accumulate in cancer cells, resulting in abnormal activation of signal transduction pathways and driving malignant processes (Science. 2002, 298, 1912-1934). Pharmacological inhibition of these signal transduction pathways presents promising intervention opportunities for targeted cancer therapy (Nature. 2004, 432, 294-297).

[0003] The mesenchymal epithelial transition factor (Met) gene is located on the long arm of human chromosome 7 and contains 21 exons. c-Met is a receptor tyrosine kinase (RTKs) with autonomous phosphorylation activity encoded by the Met gene. c-Met and its only known ligand HGF (hepatocyte growth factor) play an important role in cell proliferation, survival, invasion, tissue development and organ regeneration. Abnormal forms of the Met gene include mutation, amplification, rearrangement and overexpression. Met rearrangement is very rare in lung cancer. In lung cancer, the probability of c-Met protein overexpression is 33.6%, the probability of c-Met gene amplification is 9.8-20.0%, and the probability of c-Met mutation is 0.8-4.0%. At present, drugs for Met or HGF targeted therapy are mainly divided into two categories: small molecule kinase inhibitors and monoclonal antibodies. Small molecule kinase inhibitors can be further divided into multikinase inhibitors (crizotinib, cabozantinib, MGCD265, AMG208, Altiratinib, and Golvatinib, etc.) and selective Met inhibitors (competitive ATP inhibitors: Capmatinib and Tepotinib [MSC2156119J]; noncompetitive ATP inhibitors: tivantinib). Monoclonal antibodies can be further divided into anti-MET antibodies (onartuzumab and emibetuzumab [LY2875358]) and anti-HGF antibodies (ficlatuzumab [AV-299] and rilotumumab [AMG102]).

[0004] SFK is a member of the SRC family and is a cytoplasmic tyrosine kinase that plays an important role in cell signal transduction induced by extracellular stimuli (including growth factors and integrins) (Oncogene, 2004, 23, 7906-7909). At present, researchers have found and reported the increase in expression of the non-receptor tyrosine kinase SRC and / or the increase in SRC kinase activity in a variety of human cancers, including breast cancer, colon cancer, lung cancer and head and neck cancer. It is reported that the increase in SRC expression / activity and the activation of its downstream STAT3 are associated with a variety of epithelial cancers and are associated with the expression of a variety of growth factors such as vascular endothelial growth factor and HGF. SRC is a key downstream transducer of Met-driven tumor growth. SRC activation is essential for the ligand-dependent and ligand-independent activation of Met. In Met-driven gastric cancer cell lines, SRC inhibition increased the sensitivity of cells to c-Met inhibition, providing a theoretical basis for the combined treatment of c-Met inhibitors and SRC inhibitors (Clin Cancer Res. 2010, 16, 3933-3943). Although HGF / Met signaling is associated with the development and progression of colorectal cancer (CRC), the therapeutic effect of Met inhibitors alone is not obvious. In mutant and wild-type RAS cells, combined inhibition of Met and SRC increased the inhibition of cell proliferation and apoptosis (Exp Ther Med. 2017.4692).

[0005] CSF1R (colony-stimulating factor 1 receptor, CSF1R) is a colony stimulating factor 1 receptor. Colony stimulating factor (CSF1) is a cytokine that controls the production, differentiation and function of macrophages. Uncontrolled inflammation in the tumor microenvironment is a hallmark of cancer and is associated with M2 polarized macrophages. Tumor-associated macrophages (TAMs) are more similar to M2 polarized macrophages and play an important role in promoting the proliferation, invasion and metastasis of cancer (J Hematol Oncol. 2017, 10, 58). The tumor-promoting function of TAMs is based on their ability to secrete pro-angiogenic and growth factors and their ability to strongly inhibit T cell effector function by releasing immunosuppressive cytokines and affecting T cell metabolism (Cancer Cell. 2014, 25, 846-859). Although anti-PD-1 monoclonal antibodies (mAbs) targeting immune checkpoints have shown benefits in treating certain cancers, these drugs do not remain effective. In patients with advanced solid tumors, survival of TAMs is mediated by signaling through CSF1R, and inhibition of CSF1R signaling decreases TAMs and increases CD8 / CD4 +T cell ratio. Therefore, for solid tumors, whether as a monotherapy or combined drug therapy, targeting CSF1R signaling that causes TAM regulation is a promising therapeutic strategy. Co-expression of CSF1R and CSF1 is most often detected in invasive tumors. Activation of CSF1R promotes Src-dependent destruction of the breast epidermal architecture, indicating that targeted inhibition of CSF-1R and SRC is likely to be a valuable strategy for the treatment of invasive tumors. Tenosynovial giant cell tumor (TGCT) or pigmented villonodular synovitis (PVNS) is a clonal neoplastic proliferation produced by cells overexpressing CSF1, which recruits polyclonal macrophages loaded with CSF1R and constitutes the main body of the tumor. The use of CSF1R small molecule inhibitors has an ameliorative effect on the affected junctions (Curr Opin Oncol. 2011, 23, 361-366).

[0006] In summary, the inhibition of multiple kinases such as MET / SRC / CSF1R has great potential for the treatment of cancer. However, there are no clinically available compounds that inhibit Met / Src and / or CSF1R. The MET / SRC / CSF1R and other multiple kinases inhibitors of the invention have good brain penetration effects and will significantly make up for unmet clinical needs. Summary of the invention

[0007] One object of the present invention is to provide a novel compound or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled compound, isomer or prodrug thereof, which has excellent tyrosine kinase inhibitory activity.

[0008] Another object of the present invention is to provide a pharmaceutical composition.

[0009] Another object of the present invention is to provide uses of the novel compound or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof.

[0010] The present application provides a compound as shown in formula (I) or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled compound, isomer or prodrug thereof,

[0011]

[0012] Wherein, X1 is selected from -O-, -S- or -NR 11 -;

[0013] X2 is selected from -CH2-, -O-, -S- or -NH-;

[0014] X3 is selected from O, S or NR 10 ;

[0015] Y1 and Y2 are both different and selected from C or N;

[0016] The dotted line in the ring indicates the presence of conjugated double bonds in the ring;

[0017] R1, R4, R5, R6, R 10 , R 11 are each independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～8 Alkyl, deuterated C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Alkylamino, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano;

[0018] R2 and R3 are each independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl, cyano; or the C atom to which it is connected and the X2 group together form a 3-10-membered cycloalkyl, a 3-10-membered heterocyclic group containing at least one heteroatom, or a 5-10-membered heteroaryl containing at least one heteroatom;

[0019] Alternatively, when X1 is selected from -NR 11 -, the N atom and the C atom in CR2R3 and together with R 11 Together with R2, it forms a 3-10-membered nitrogen heterocycloalkyl group;

[0020] R 2’ , R 3’ are each independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20Heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl, cyano; or together with the connected C and adjacent N atoms, form a 3-10 membered heterocyclic group containing at least one heteroatom or a 5-10 membered heteroaryl group containing at least one heteroatom;

[0021] m and n represent integers of 1 to 10; wherein when Y1=C, Y2=N, X3=O, X2=NH and R6=H, m represents an integer of 3 to 10;

[0022] The substituents of the above mentioned groups may be selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano.

[0023] Furthermore, in the above formula (I), R1, R4, R5, R6, R 10 , R2, R3, R 2’ , R 3’ The optional substituents thereof may represent groups including, but not limited to, hydrogen, deuterium, fluorine, chlorine, bromine, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, -CN, -CF3, -NH2, -NH(C 1～4 Alkyl), -N(C 1～4 Alkyl)2, -CO2C 1～4 Alkyl, -CO2H, -NHC(O)C 1～4 Alkyl, -SO2C 1～4 Alkyl, -C(O)NH2, -C(O)NH(C 1～4 alkyl), -C(O)N(C 1～4 alkyl) 2, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, pyrrolidinyl, pyrazolyl, piperidinyl, pyridyl, piperazinyl, triazinyl, furanyl, thiofuranyl, morpholinyl, thiomorpholinyl, phenyl, naphthyl, biphenyl, terphenyl and the like.

[0024] In one embodiment, the compound has the structure of the following formula I-1 or I-2:

[0025]

[0026] Wherein, the definitions of each group are as described above;

[0027] Furthermore, when X2 is NH, X3=O, and m=1-2, R6 is not H.

[0028] In one embodiment, the compound has the structure of the following formula I-11, preferably formula I-12 or I-13;

[0029]

[0030] Wherein, the definitions of each group are as described above,

[0031] In formulas I-12 and I-13, R7 is independently selected from the following groups which may be substituted or unsubstituted: hydrogen, halogen, C 1～8 Alkyl, deuterated C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Alkylamino, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano; m' represents an integer of 1-10.

[0032] In one embodiment, the compound has the structure of the following formula I-21, preferably formula I-22 or I-23;

[0033]

[0034] Wherein, the definitions of each group are as described above,

[0035] In formulas I-22 and I-23, R7 is independently selected from the following groups which may be substituted or unsubstituted: hydrogen, halogen, C 1～8 Alkyl, deuterated C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Alkylamino, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano; m' represents an integer of 1-10.

[0036] On the other hand, the present application also provides a compound as shown in formula (II) or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof,

[0037]

[0038] Wherein, X1 is selected from -O-, -S- or -NR11 -;

[0039] X2 is selected from -CH2-, -O-, -S- or -NH-;

[0040] X3 is selected from O, S or NR 10 ;

[0041] Y1 and Y2 are both different and selected from C or N;

[0042] The dotted line in the ring indicates the presence of conjugated double bonds in the ring;

[0043] R4, R5, R6, R 10 , R 11 are each independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～8 Alkyl, deuterated C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Alkylamino, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano;

[0044] R2 and R3 are each independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl, cyano; or the C atom to which it is connected and the X2 group together form a 3-10-membered cycloalkyl, a 3-10-membered heterocyclic group containing at least one heteroatom, or a 5-10-membered heteroaryl containing at least one heteroatom;

[0045] Alternatively, when X1 is selected from -NR 11 -, the N atom and the C atom in CR2R3 and together with R 11 Together with R2, it forms a 3-10-membered nitrogen heterocycloalkyl group;

[0046] R 2’ , R 3’ are each independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl, cyano; or together with the connected C and adjacent N atoms, form a 3-10 membered heterocyclic group containing at least one heteroatom or a 5-10 membered heteroaryl group containing at least one heteroatom;

[0047] R8 and R9 are each independently selected from halogen;

[0048] m and n represent integers from 1 to 10;

[0049] The substituents of the above mentioned groups may be selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano.

[0050] The compound has the following structure of formula II-1 or II-2:

[0051]

[0052] Wherein, the definitions of each group are as described above.

[0053] In one embodiment, the compound has the structure of the following formula II-11, preferably formula II-12 or II-13;

[0054]

[0055] Wherein, the definitions of each group are as described above,

[0056] In formula II-12 and II-13, R7 is independently selected from the following groups which may be substituted or unsubstituted: hydrogen, halogen, C 1～8 Alkyl, deuterated C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Alkylamino, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano; m' represents an integer of 1-10.

[0057] In one embodiment, the compound has the structure of the following formula II-21, preferably formula II-22 or II-23;

[0058]

[0059] Wherein, the definitions of each group are as described above,

[0060] In formula II-22 and II-23, R7 is independently selected from the following groups which may be substituted or unsubstituted: hydrogen, halogen, C 1～8 Alkyl, deuterated C 1～8 Alkyl, C 1～8 Alkoxy, C 1～8 Alkylamino, C 1～8 Haloalkyl, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, hydroxyl, thiol, carboxyl, ester, acyl, amino, amide, sulfonyl or cyano; m' represents an integer of 1-10.

[0061] In one embodiment, R1 is F; or, R9 is F.

[0062] In one embodiment, R5 is selected from C 1～8 Alkyl, C 1～8 Haloalkyl, deuterated C 1～8 Alkyl, C 3～8 Cycloalkyl C 1～8 Alkyl or cyano C 1～8 Alkyl; preferably ethyl, deuterated ethyl, cyclopropylmethyl or cyanomethyl.

[0063] In one embodiment, R4 is hydrogen or amino.

[0064] In one embodiment, R6 is selected from hydroxyl, amino, C 1～8 Alkoxy or C 1～8 Preferably, R6 is hydroxyl; or, R6 is amino; or, R6 is C 1～8 Alkoxy; or, R6 is C 1～8 Alkylamino.

[0065] In one embodiment, X1 is -O-.

[0066] In one embodiment, X2 is -O-; or, X2 is -NH-.

[0067] In one embodiment, X3 is -O-; or, X3 is NR 10 , R 10 Selected from hydroxyl or C1～8 Alkoxy.

[0068] In one embodiment, m, n and m′ may represent an integer of 1, 2, 3, 4, 5 or 6, in particular 1, 2 or 3.

[0069] In one embodiment, in the above formulae, when the substituents R2 and R3 on the C atom are independently selected from the group forming a cycloalkyl, heterocyclyl or heteroaryl group together with the X2 group, it refers to a -(CR2R3)- group adjacent to the X2 group, wherein R2 or R3 forms together with the C to which it is connected and the X2 group.

[0070] In one embodiment of the present invention, in the above formulas, when R2 and R3 are each independently a single bond connecting the C atom and the adjacent macrocyclic ring atom, the adjacent macrocyclic ring atom may be a ring atom C in another -(CR2R3)- group adjacent thereto, or may be a ring atom in the X2 group adjacent thereto. In a preferred embodiment, R2 and R3 in the -(CR2R3)- group adjacent to the X2 group may each independently be a single bond connecting the C atom and the X2 group, that is, R2 and / or R3 are single bonds connecting the C atom and the X2 central atom. For example, when one of R2 and R3 is a single bond, the X2 group forms a double bond with the adjacent -(CR2R3)- group; when two of R2 and R3 are single bonds, the X2 group forms a triple bond with the adjacent -(CR2R3)- group.

[0071] Similarly, substituted or unsubstituted double bonds or triple bonds can be formed between two adjacent -(CR2R3)- groups or between C and C atoms in the macrocycle.

[0072] When X1 is selected from -NR 11 -, the N atom and the C atom in CR2R3 and together with R 11 Together with R2, it can form a 3- to 10-membered azacycloalkyl group, for example, azetidinyl.

[0073] In one embodiment of the present invention, in the above formulae, the substituents R2 and R3 on each C atom are independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～5 Alkyl, C 1～5 Alkoxy, C 1～5 Haloalkyl, C 3～6 Cycloalkyl or R2, R3 are each independently a single bond connecting the C atom and the adjacent macrocyclic ring atom.

[0074] In one embodiment of the present invention, in the above formulas, R 2’ , R 3’are each independently selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～5 Alkyl, C 1～5 Alkoxy, C 1～5 Haloalkyl, C 3～6 Cycloalkyl, or together with the connected C and L2, forms a 4-8 membered heterocyclic group containing at least one heteroatom.

[0075] In one embodiment according to the present invention, in the above formulae, the -CR2R3- group or -CR 2’ R 3’ The macrocyclic atom C in the -group or the C in the substituent may generate one or more chiral centers depending on the group. The present invention includes all optical isomers and racemates.

[0076] In one embodiment according to the present invention, in the above formulae, R4 is selected from the following substituted or unsubstituted groups: hydrogen, halogen, C 1～5 Alkyl, C 1～5 Alkoxy, C 1～5 Haloalkyl, C 3～6 Cycloalkyl, hydroxy, mercapto, carboxyl, amino or cyano.

[0077] In one embodiment of the present invention, the optional substituents in the above embodiment are selected from fluorine, bromine, -CN, -OH, -CF3, -NH2, -NH(C 1～4 Alkyl), -N(C 1～4 Alkyl)2, -CO2C 1～4 Alkyl, -CO2H, -NHC(O)C 1～4 Alkyl, -SO2C 1～4 Alkyl, -C(O)NH2, -C(O)NH(C 1～4 alkyl), -C(O)N(C 1～4 Alkyl)2, C 1～5 Alkyl, C 3～6 Cycloalkyl, C 3～6 Heterocyclic group, C 6～10 Aryl or C 5～10 Heteroaryl.

[0078] In one embodiment according to the present invention, the compound is selected from the following compounds:

[0079]

[0080]

[0081] The present invention also provides a pharmaceutical composition, comprising the compound described in any one of the above technical solutions or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and a pharmaceutically acceptable carrier.

[0082] The pharmaceutical composition includes, but is not limited to, oral dosage forms, parenteral dosage forms, external dosage forms, rectal dosage forms, etc. For example, the pharmaceutical composition can be oral tablets, capsules, pills, powders, sustained-release preparations, solutions and suspensions, sterile solutions, suspensions or emulsions for parenteral injection, ointments, creams, gels, etc. for external use, or suppositories for rectal administration.

[0083] The pharmaceutical composition may also include other active ingredients or drugs for combined use with the compound or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug.

[0084] The present invention also provides the use of the above-mentioned compound or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope label, isomer or prodrug, and the above-mentioned pharmaceutical composition in the preparation of a drug for treating a tyrosine kinase-mediated disease; and a method for treating a tyrosine kinase-mediated disease, comprising administering an effective amount of the above-mentioned compound or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope label, isomer or prodrug, or the above-mentioned pharmaceutical composition to a patient.

[0085] Furthermore, the tyrosine kinase is selected from one or more of the following: SRC, MET, CSF1R, ALK, ROS1, TRKA, TRKB, TRKC, JAK2, SRC, FYN, LYN, YES, FGR, FAK, AXL, ARK5.

[0086] Furthermore, the tyrosine kinase-mediated diseases include cancer, pain, neurological diseases, autoimmune diseases and inflammation.

[0087] Furthermore, the tyrosine kinase-mediated cancers may include lung cancer, colorectal cancer, breast cancer, ovarian cancer, thyroid cancer, prostate cancer, hepatocellular carcinoma, renal cell carcinoma, gastric and esophageal cancer, bile duct cancer, glioma, glioblastoma, head and neck cancer, inflammatory myofibroblastic tumor, angiosarcoma, epithelioid hemangioendothelioma, anaplastic large cell lymphoma, etc.

[0088] Furthermore, the tyrosine kinase-mediated pain can be pain of any origin or etiology, including cancer pain, chemotherapy pain, neuralgia, injury pain or other origins.

[0089] Furthermore, the tyrosine kinase-mediated autoimmune diseases include rheumatoid arthritis, Sjogren's syndrome, type I diabetes, lupus, and the like.

[0090] Furthermore, the tyrosine kinase-mediated neurological diseases include Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, and the like.

[0091] Furthermore, the tyrosine kinase-mediated inflammatory diseases include atherosclerosis, allergy, inflammation caused by infection or injury, and the like.

[0092] The present application also provides a combination drug for treating cancer in a patient, which contains a therapeutically effective amount of a preparation for inhibiting SRC, MET and / or CSF1R and an additional anticancer agent, which are administered simultaneously or separately in the same or different specifications, wherein the preparation for inhibiting SRC, MET and / or CSF1R comprises the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof, or the pharmaceutical composition according to claim 16; the additional anticancer agent is an EGFR antibody or an EGFR small molecule inhibitor, preferably selected from cetuximab, naximol, panitumumab or amivantamab dual antibody, afatinib, brigatinib, canertinib, dacomitinib, erlotinib, gefitinib, HKI 357, lapatinib, osimertinib, naquatinib, nazartinib, neratinib, omotinib, pelitinib, PF-06747775, rociletinib, vandetanib, ametinib, vometinib, Mobocertinib, DZD9008, BEBT-109, lazertinib, CLN-081, WTS-004, JFAN-1001, C-005, XZP-5809-TT1, JRF103, FWD1509, JNJ-372, or a pharmaceutically acceptable salt thereof.

[0093] The diaryl macrocyclic compound and its pharmaceutical composition provided by the present invention have significant tyrosine kinase inhibitory activity, can overcome tumor drug resistance, can break through the blood-brain barrier, have excellent pharmacokinetic properties and excellent oral bioavailability, can be administered in a smaller dose, thereby reducing the patient's treatment cost and possible toxic side effects, and thus have great application potential. Moreover, the inventors of the present application have found that when a cyano group (-CN) exists at the ortho position on the benzene ring of the macrocyclic compound and the amino hydrogen in the ring is replaced by an alkyl or other group, it can have a significant effect on the activity of the compound, such as increasing the kinase inhibitory activity of SRC, MET, and CSF1R and improving the selectivity for other kinases. The inventors of the present application have also found that when two halogen groups exist at the ortho and meta positions on the benzene ring of the macrocyclic compound, especially when the meta position is F, it can also have a significant effect on the activity of the compound, such as increasing the kinase inhibitory activity of SRC, MET, and CSF1R. The inventors of the present application have found that when some groups of the macrocyclic compound are comprehensively changed, the permeability of the compound is significantly improved, and the blood-brain barrier permeability is increased. BRIEF DESCRIPTION OF THE DRAWINGS

[0094] Figure 1 The bar graph shows the average concentration of the compound in Example 1 in rat brain tissue and plasma after oral administration.

[0095] Figure 2 The bar graph shows the average concentration of TPX-0022 in rat brain tissue and plasma after oral administration DETAILED DESCRIPTION

[0096] definition

[0097] In this application, each group may have the following definitions:

[0098] Hydrogen can be represented as -H, and can also be replaced by isotopes such as deuterium and tritium.

[0099] Halogen may include fluorine, chlorine, bromine, iodine.

[0100] C 1～8 The alkyl group may include methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, heptyl, octyl, and the like.

[0101] Deuterated C 1～8 Alkyl, tritiated C 1～8 Alkyl can represent the C 1～8 One or more or even all of the hydrogen atoms on the alkyl group are replaced by isotopes such as deuterium and tritium.

[0102] C 1～8 Alkoxy can be represented by -OC 1～8 Alkyl, where C 1～8 Alkyl groups include those defined above; for example, C 1～8 The alkoxy group may include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy and the like.

[0103] C 1～8 The alkyl halide can be represented by C 1～8 Any number of hydrogen atoms in the alkyl group are replaced by halogen, where C 1～8 The groups including alkyl and halogen are as defined above; for example, C 1～8 The haloalkyl group may include -CF3 and the like.

[0104] C 3～8 Cycloalkyl can be represented by a non-aromatic saturated carbocyclic ring, including monocarbocyclic rings (having one ring) and dicarbocyclic rings (having two rings), for example, C 3～8 The cycloalkyl group may include wait.

[0105] C 3～8 Cycloalkyl C 1～8 Alkyl groups can be represented by 3～8 Cycloalkyl C 1～8 Alkyl, where C 3～8 Cycloalkyl and C 1～8 The definition of alkyl is as above, for example, C 3～8 Cycloalkyl C 1～8 The alkyl group may include cyclopropylmethyl, cyclobutylmethyl, cyclohexylethyl, and the like.

[0106] C 3～8 Heterocyclic groups can be represented by C 3～8 A group obtained by replacing any number of ring atoms in a cycloalkyl group with heteroatoms such as O, S, N, P, or Si, wherein C 3～8 The groups included in the cycloalkyl group are as defined above. For example, C 3～8The heterocyclic group can include oxirane, thioethanethiol, aziridinyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, pyrrolidinyl, oxazolidinyl, tetrahydropyrazolyl, pyrrolinyl, dihydrofuranyl, dihydrothiophenyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyranyl, tetrahydropyranyl, dihydrothiopyranyl, azepanyl, oxetanyl, thiepanyl, oxazabicyclo[2.2.1]heptyl, azaspiro[3.3]heptyl, and the like.

[0107] C 6～20 The aryl group may include a monocyclic aryl group, a bicyclic aryl group or a polycyclic aryl group, for example, may include phenyl, biphenyl, naphthyl, phenanthrenyl, anthracenyl, azulenyl and the like.

[0108] C 5～20 Heteroaryl can represent an unsaturated group containing any number of heteroatoms such as O, S, N, P, Si as ring atoms. 5～20 Heteroaryl groups may include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, quinolyl, isoquinolyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, and the like.

[0109] The hydroxyl group can be represented as -OH.

[0110] A thiol group can be represented as -SH.

[0111] The carboxyl group can be represented as -COOH.

[0112] The ester group can be represented by -COOR', and the definition of R' can be the definition of the substituent described in formula (1), for example, C 1～8 Alkyl substituted ester group, represented by -COOC 1～8 Alkyl, where C 1～8 Alkyl groups include groups as defined above.

[0113] The acyl group can be represented by -COR', and the definition of R' can be the definition of the substituent described in formula (1), for example, C 1～8 Alkyl substituted acyl, represented by -COC 1～8 Alkyl, where C 1～8 Alkyl groups include groups as defined above.

[0114] The amino group can be represented by -NH2, -NHR' or -N(R')2, where R' can be defined as the substituent described in formula (1), for example, C 1～8 Alkyl-substituted amino groups can be represented by -NHC 1～8 Alkyl or -N(C 1～8 Alkyl)2, where C 1～8Alkyl groups include groups as defined above.

[0115] The amide group may be represented by -COamino wherein the amino group is as defined above.

[0116] The sulfonyl group can be represented by -S(O)2R', where R' can be defined as the substituent group described in formula (1), for example, C 1～8 Alkyl substituted sulfonyl, represented by -S(O)2C 1～8 Alkyl, where C 1～8 Alkyl groups include groups as defined above.

[0117] The cyano group can be represented as -CN.

[0118] In the above definition, when the number of carbon atoms changes, the above definition only changes according to the change in the number of carbon atoms and does not affect the definition of the group type; for example, "C 1～5 The "alkyl" may include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, etc. 1～8 The term "alkyl" refers to any group having 1 to 5 carbon atoms.

[0119] Details

[0120] Unless otherwise defined, all technical terms herein have the same meaning as those commonly understood by those skilled in the art to which the subject matter of the claims belongs. Unless otherwise indicated, all patents, patent applications, and public materials cited herein are incorporated herein by reference in their entirety. When a trade name appears in this document, it is intended to refer to the corresponding commodity or its active ingredient.

[0121] It should be understood that the above brief description and the following detailed description are exemplary and are only used for explanation, and do not limit the subject matter of the invention herein. In this application, it must be noted that unless otherwise clearly stated in the text, the singular forms used in this specification and claims include the plural forms of the referred things. It should also be noted that unless otherwise stated, the use of "or" and "or" means "and / or". In addition, the use of the term "including" and other forms, such as "including", "containing" and "containing" are not restrictive.

[0122] Definitions of standard chemical terms can be found in the literature, including Advanced Organic Chemistry by Carey and Sundberg. thEd, Vol A (2000) and B (2001), Plenum Press, New York. Unless otherwise indicated, conventional methods within the skill of the art are used, such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacological methods. Unless specifically defined, the nomenclature and laboratory operations and techniques in analytical chemistry, organic synthetic chemistry, and medical and pharmaceutical chemistry are known to those skilled in the art. Standard techniques can be used for chemical synthesis, chemical analysis, drug preparation, formulation, drug delivery and patient treatment. Standard techniques can be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipid infection). For example, the reaction and purification techniques can be performed using a kit with instructions provided by the manufacturer, or according to methods known in the art, or according to the methods described in the present invention. In general, the aforementioned techniques and steps can be performed by conventional methods well known in the art and described in various general or more specific literature, which are cited and discussed in the present invention.

[0123] When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result if the formula were written from right to left. For example, CH2O is equivalent to OCH2.

[0124] The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, as long as the valence state of the specific atom is normal and the substituted compound is stable. When the substituent is an oxo (i.e., =O), it means that two hydrogen atoms are replaced, and the oxo will not occur on an aromatic group.

[0125] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice. In addition, combinations of substituents and / or variants thereof are permitted only if such combinations result in stable compounds.

[0126] The C m～n means that the part has m to n carbon atoms. 1～8 "A group means that the part has 1-8 carbon atoms, that is, the group contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms...8 carbon atoms. Therefore, for example, "C 1～8"Alkyl" refers to an alkyl group containing 1-8 carbon atoms, that is, the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl...octyl, etc. The numerical ranges herein, such as "1-8", refer to each integer in the given range, such as "1-8 carbon atoms" means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms or 8 carbon atoms.

[0127] The term "membered" refers to the number of backbone atoms that make up the ring. For example, pyridine is a six-membered ring and pyrrole is a five-membered ring.

[0128] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0129] The term "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical component or agent, namely a "carrier" that facilitates the introduction of the compound into cells or tissues, including but not limited to stabilizers, diluents, suspending agents, thickening agents and / or excipients.

[0130] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acid and free base of the specified compound and has no adverse effects in biology or other aspects. Unless otherwise indicated, the salts in the present invention may refer to metal salts, ammonium salts, salts formed with organic bases, salts formed with inorganic acids, salts formed with organic acids, salts formed with basic or acidic amino acids, etc. Non-limiting examples of metal salts include, but are not limited to, salts of alkali metals, such as sodium salts, potassium salts, etc.; salts of alkaline earth metals, such as calcium salts, magnesium salts, barium salts, etc.; aluminum salts, etc. Non-limiting examples of salts formed with organic bases include, but are not limited to, salts formed with trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, etc. Non-limiting examples of salts formed with inorganic acids include, but are not limited to, salts formed with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc. Non-limiting examples of salts formed with organic acids include, but are not limited to, salts formed with formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, maleic acid, tartaric acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc. Non-limiting examples of salts formed with basic amino acids include, but are not limited to, salts formed with arginine, lysine, ornithine, etc. Non-limiting examples of salts formed with acidic amino acids include, but are not limited to, salts formed with aspartic acid, glutamic acid, etc.

[0131] Pharmaceutically acceptable salts can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. In general, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of an appropriate base or acid in water or an organic solvent or a mixture of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.

[0132] The term "solvate" refers to a physical aggregate of a compound of the present invention and one or more solvent molecules, which includes varying degrees of ionic and covalent bonds, such as hydrogen bonds. It has been shown that this solvate can be separated, for example, when one or more solvent molecules are mixed in the lattice of the crystal. "Solvate" includes two parts: a solvent phase and a separable solvate. There are many examples of corresponding solvates, including ethanol solvates, methanol solvates, etc. "Hydrate" is a solvate with water (H2O) molecules as solvent. One or more compounds of the present invention can be prepared into solvates at will. The preparation of solvates is well known. For example, M.Caira et al, J.Pharmaceutical Sci., 93 (3), 601-611 (2004) describes the preparation of a solvate of the antifungal drug fluconazole, which is prepared using ethyl acetate and water. EC van Tonder et al, AAPS Pharm Sci Tech., 5 (1), article 12 (2004); and AL Bingham et al, Chem. Commun., 603-604 (2001) also describe similar preparation methods for solvates and hydrates. A typical, non-limiting preparation process is to dissolve the compound of the invention in a desired amount of an ideal solvent (organic solvent or water or a mixed solvent thereof) at a temperature higher than room temperature, cool it, place it for crystallization, and then separate and pick out the crystals using a standard method. The presence of the solvent (water) that forms the solvate (hydrate) in the crystals can be confirmed by IR spectroscopy analysis technology.

[0133] The term "active metabolite" refers to an active derivative of a compound that is formed when the compound is metabolized.

[0134] The term "polymorph" refers to compounds of the invention that exist in different crystalline lattice forms.

[0135] The term "isotope-labeled compound" refers to a compound of the present invention that is isotopically labeled. For example, the isotopes in the compound of the present invention may include various isotopes of elements such as H, C, N, O, P, F, S, etc. 2 H, 3 H, 13 C, 14 C, 15 N,18 Oh, 17 Oh, 31 P, 32 P, 35 S, 18 F and 36 S.

[0136] The term "pharmaceutically acceptable prodrug" or "prodrug" refers to any pharmaceutically acceptable salt, ester, salt of an ester, or other derivative of a compound of the invention, which, upon administration to a receptor, is capable of providing, directly or indirectly, a compound of the invention or a pharmaceutically active metabolite or residue thereof. Particularly preferred derivatives or prodrugs are those compounds that, when administered to a patient, can increase the bioavailability of the compound of the application (e.g., can make an orally administered compound more easily absorbed into the blood), or those compounds that promote the delivery of the parent compound to a biological organ or site of action (e.g., the brain or lymphatic system). Prodrugs can be prepared by modifying functional groups present in the compound in a manner that allows for decomposition into the parent compound, either by conventional manipulation or in vivo. Various prodrug forms are well known in the art. See, for a discussion of prodrugs, Pro-drugs as Novel Delivery Systems, by T. Higuchi and V. Stella (1987) Vol. 14 of the ACSSymposium Series, Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and in Pergamon Press. Design of Prodrugs, Bundgaard, A. Ed., Elseview, 1985 and Method in Enzymology, Widder, K. et al., Ed.; Academic, 1985, vol. 42, p. 309-396; Bundgaard, H. "Design and Application of Prodrugs" in A Textbook of Drug Design and Development, Krosgaard-Larsen and H. Bundgaard, Ed., 1991, Chapter 5, pp. 113-191; and Bundgaard, H., Advanced Drug Delivery Review, 1992, 8, 1-38, the above documents are incorporated herein by reference.

[0137] The term "stereoisomer" refers to an isomer resulting from different spatial arrangements of atoms in a molecule. The compounds of the present invention contain structures such as asymmetric or chiral centers, double bonds, etc. Therefore, the compounds of the present invention may include multiple isomeric forms such as optical isomers, geometric isomers, tautomers, atropisomers, etc. These isomers and their single isomers, racemates, etc. are all included in the scope of the present invention. For example, for optical isomers, optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral resolution, chiral synthesis or chiral reagents or other conventional techniques. For example, it can be converted into diastereomers by reaction with appropriate optically active substances (such as chiral alcohols or Mosher's acyl chlorides), which are separated and converted (such as hydrolyzed) into corresponding single isomers. For another example, separation can also be carried out by a chromatographic column.

[0138] The "pharmaceutical composition" herein can be prepared in a manner well known in the pharmaceutical art, and can be administered or applied by a variety of routes, depending on whether local or systemic treatment is required and the area to be treated. It can be administered topically (e.g., transdermal, skin, eye and mucous membrane including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, such as intrathecal or intraventricular administration. It can be administered parenterally in the form of a single bolus, or it can be administered, for example, by a continuous infusion pump. The pharmaceutical compositions herein include, but are not limited to, the following forms: tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or dissolved in a liquid vehicle); ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders, etc.

[0139] The pharmaceutical composition herein can be formulated in unit dosage form, each dose can contain about 0.1 to 1000 mg, usually about 5 to 1000 mg of active ingredient, more usually about 100 to 500 mg of active ingredient. The term "unit dosage form" refers to physically discrete single dosage units suitable for use in human patients and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect in admixture with a suitable pharmaceutical carrier.

[0140] The term "subject" refers to an individual suffering from a disease, disorder, condition, etc., including mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals, such as rabbits, dogs and cats; laboratory animals, including rodents, such as rats, mice and guinea pigs, etc.

[0141] The term "treat" and other similar synonyms include relieving, alleviating or ameliorating symptoms of a disease or condition, preventing other symptoms, ameliorating or preventing the underlying metabolic causes of symptoms, inhibiting a disease or condition, such as preventing the development of a disease or condition, alleviating a disease or condition, making a disease or condition better, alleviating symptoms caused by a disease or condition, or stopping symptoms of a disease or condition, and in addition, the term may also include the purpose of prevention. The term also includes obtaining a therapeutic effect and / or a preventive effect. The therapeutic effect refers to curing or improving the underlying disease being treated. In addition, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, for example, although the patient may still be affected by the underlying disease, the patient's condition is observed to improve. In terms of preventive effects, the composition or compound can be administered to a patient at risk for a particular disease, or even if a disease diagnosis has not yet been made, the composition or compound can be administered to a patient who has one or more physiological symptoms of the disease.

[0142] The term "amount necessary to achieve the therapeutic effect" or "therapeutically effective amount" refers to the amount of at least one agent or compound that is sufficient to relieve to some extent one or more symptoms of the disease or condition being treated after administration. The result can be a reduction and / or alleviation of signs, symptoms or causes, or any other desired change in a biological system. Techniques such as dose escalation tests can be used to determine the effective amount suitable for any individual case. The actual amount of compound, pharmaceutical composition or medicament administered is usually determined by the physician based on relevant circumstances, including the condition being treated, the route of administration selected, the actual compound administered; the age, weight and response of the individual patient; the severity of the patient's symptoms, etc.

[0143] The ratio or concentration of the compound of the present invention in the pharmaceutical composition may not be fixed, depending on a variety of factors, including dosage, chemical properties (e.g., hydrophobicity), route of administration, etc. For example, the compound of the present invention can be provided by a physiologically buffered aqueous solution containing about 0.1 to 10% w / v of the compound for parenteral administration. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg body weight / day. In certain embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg body weight / day. The dosage is likely to depend on such variables as the type and extent of the disease or condition, the general health status of the particular patient, the relative biological efficacy of the selected compound, the excipient formulation, and its route of administration.

[0144] The term "administering" refers to a method that enables a compound or composition to be delivered to a desired site for a biological effect. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injections or infusions), topical and rectal administration. Those skilled in the art are familiar with administration techniques that can be used for the compounds and methods described herein, such as those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.

[0145] The term "IC 50 ” means a 50% inhibition of the maximal effect in the assay measuring such effect.

[0146] In order to make the objectives, technical solutions and advantages of the present invention more clear, the technical solutions of exemplary embodiments of the present invention will be further described below.

[0147] The present invention can prepare the compounds described in the present invention by the following methods. The following methods and examples are intended to illustrate these methods. These processes and examples should not be construed as limiting the present invention in any way. The compounds described herein can also be synthesized using standard synthetic techniques known to those skilled in the art, or a combination of methods known in the art and the methods described herein.

[0148] The chemical reactions of the embodiments of the present invention are carried out in a suitable solvent, which must be suitable for the chemical changes of the present invention and the reagents and materials required. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing implementation methods.

[0149] An important consideration in any synthetic route planning in this area is to select a suitable protecting group for a reactive functional group (such as the amino group in the present invention). For trained practitioners, Greene and Wuts (Protective Groups In Organic Synthesis, Wiley and Sons, 1991) is an authority in this regard. All references cited in the present invention are incorporated herein in their entirety.

[0150] The reactions described herein can be monitored by any suitable method known in the art. For example, the reactions can be monitored by broad spectrum methods such as nuclear magnetic resonance spectroscopy (e.g. 1H or 13 C), infrared spectroscopy, spectrophotometry (eg, UV-visible), mass spectroscopy, or the like, or by chromatography, such as high performance liquid chromatography (HPLC) or thin layer chromatography, to monitor product formation.

[0151] Preparation of intermediates

[0152] Intermediate 1: tert-butyl 2-amino-5-(p-toluenesulfonyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate (Compound I1)

[0153]

[0154] Step A: (Z)-3-Amino-4,4,4-trichloro-2-cyano-butenoic acid tert-butyl ester

[0155]

[0156] At 0°C, a catalytic amount of triethylamine (30 mL, 0.2 mol) was added dropwise to a solution of tert-butyl cyanoacetate (500 g, 3.5 mol) and trichloroacetonitrile (895 g, 6.2 mol) in ethanol (1.5 L). After the addition was completed, the mixture was reacted at 0°C for 2 hours, and then slowly warmed to room temperature for 5 hours. After the reaction was completed, the solvent was removed by concentration, and the residue was stirred and slurried with a mixed solvent (PE / EA=5 / 1), and the target product (light yellow solid, 580 g, yield 57%) was obtained by filtration. The residue was concentrated and dissolved with dichloromethane, wet loaded onto a column, and purified by a chromatographic column to obtain the target product (white solid, 230 g, yield 23%).

[0157] 1 H NMR (400MHz, CDCl3) δ10.20 (brs, 1H), 6.8 (brs, 1H), 1.55 (s, 9H).m / z=285[M+1] + .

[0158] Step B: tert-Butyl 3,5-diamino-1H-pyrazole-4-carboxylate

[0159]

[0160] At room temperature, hydrazine hydrate (375 g, 6.0 mol) was slowly added dropwise to a solution (1.5 L) of (Z)-3-amino-4,4,4-trichloro-2-cyano-butenoic acid tert-butyl ester (570 g, 2.0 mol) in N,N-dimethylformamide. The system released heat significantly, and the reaction mixture was heated to 100°C and stirred for 3 hours. After cooling to room temperature, ice water was added to the system, and the mixture was extracted with ethyl acetate (×4). The organic phases were combined, washed with water (×1), washed with saturated brine (×1), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was dissolved with dichloromethane, wet loaded onto a column, and purified by a chromatographic column to obtain the target product (white solid, 391 g, yield 99%).

[0161] 1 H NMR (400MHz, DMSO-d6) δ5.29 (brs, 2H), 3.39 (brs, 2H), 1.47 (s, 9H).m / z=199[M+1] + .

[0162] Step C: tert-Butyl 2-amino-5-oxo-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carboxylate

[0163]

[0164] At room temperature, sodium ethoxide (340 g, 5.0 mol) was slowly added in batches to a solution of tert-butyl 3,5-diamino-1H-pyrazole-4-carboxylate (198 g, 1.0 mol) and 1,3-dimethylpyrimidine-2,4 (1H, 3H)-dione (140 g, 1.0 mol) in tert-butyl alcohol (1.2 L). After the addition was completed, the reaction temperature was raised to 90°C for 12 hours. After the reaction was completed, the pH value of the system was adjusted to 6 with 1N hydrochloric acid, extracted with ethyl acetate (×4), the organic phases were combined, washed with water (×1), washed with saturated brine (×1), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was stirred and slurried with a mixed solvent (PE / EA=1 / 1), and the target product (yellow solid, 175 g, 70%) was obtained by filtration.

[0165] 1 H NMR (400MHz, CDCl3) δ7.83 (d, J=8.0Hz, 1H), 5.95 (d, J=8.0Hz, 1H), 4.94 (brs, 2H), 1.62 (s, 9H).m / z=251[M+1] + .

[0166] Step D: tert-Butyl 2-amino-5-(p-toluenesulfonyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0167]

[0168] At room temperature, triethylamine (203 g, 2.0 mol) and p-toluenesulfonyl chloride (115 g, 0.6 mol) were added to a solution of tert-butyl 2-amino-5-oxo-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carboxylate (125 g, 0.5 mol) in N,N-dimethylformamide (500 mL), and the mixture was heated to 30°C for 5 hours. After cooling to room temperature, ice water was added to the residue, and the mixture was extracted with ethyl acetate (×3). The organic phases were combined, washed with water (×1), washed with saturated brine (×1), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was dissolved with dichloromethane, wet loaded onto a column, and purified by a chromatographic column to obtain the target product (pink solid, 152 g, yield 75%).

[0169] 1 H NMR (400MHz, CDCl3) δ8.32 (d, J=7.2Hz, 1H), 8.17 (d, J=8.4Hz, 2H), 7.35 (d, J=8.0H z, 2H), 6.50 (d, J=7.2Hz, 1H), 5.38 (s, 2H), 2.45 (s, 3H), 1.65 (s, 9H).m / z=405[M+1] + .

[0170] Intermediate 2: 3-bromo-2-((ethylamino)methyl)-4-fluorophenol (Compound I2)

[0171]

[0172] Step A: 2-Bromo-3,6-difluorobenzaldehyde

[0173]

[0174] At -78°C, lithium diisopropylamide (2M, 500 mL, 1.0 mol) was added to a solution of 2,5-difluorobromobenzene (150 g, 0.8 mol) in tetrahydrofuran (1.0 L), and stirred for 1 hour, followed by the addition of N,N-dimethylformamide (182 g, 2.5 mmol) at -78°C and stirring for 2 hours. The temperature was slowly raised to 0°C, and a saturated aqueous ammonium chloride solution was added at 0°C to quench the reaction mixture, followed by dilution with water and extraction with ethyl acetate (×2), the organic phases were combined, washed with water (×1), washed with saturated brine (×1), dried over anhydrous sodium sulfate, filtered and concentrated, the residue was dissolved with dichloromethane and silica gel was added to dry-mix the sample, and the target product (yellow solid, 84 g, yield 49%) was obtained by dry chromatography column purification.

[0175] 1H NMR (400MHz, CDCl3) δ10.31 (s, 1H), 7.36-7.31 (m, 1H), 7.18-7.12 (m, 1H).m / z=221[M+1] + .

[0176] Step B: 2-Bromo-3-fluoro-6-methoxybenzaldehyde

[0177]

[0178] To a solution of 2-bromo-3,6-difluorobenzaldehyde (84 g, 379 mmol) in tetrahydrofuran (200 mL) and methanol (480 mL) was added sodium methoxide (24.6 g, 455 mmol) at room temperature. The mixture was heated to 60°C and stirred for 12 hours. The reaction was completed as monitored by TLC. The reaction mixture was cooled to room temperature and concentrated under reduced pressure in vacuo to remove most of tetrahydrofuran and methanol. The reaction mixture was quenched by adding water and extracted with ethyl acetate (×2). The organic phases were combined, washed with water (×1), washed with saturated brine (×1), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was dissolved in dichloromethane and silica gel was added for dry mixing. The target product was purified by dry chromatography column (white solid, 62 g, yield 70%).

[0179] 1 H NMR (400MHz, CDCl3) δ10.30 (s, 1H), 7.20 (dd, J=7.6, 9.2Hz, 1H), 6.86 (dd, J=4.0, 9.2Hz, 1H), 3.84 (s, 3H).m / z=233[M+1] + .

[0180] Step C: 2-Bromo-3-fluoro-6-hydroxybenzaldehyde

[0181]

[0182] To a solution of 2-bromo-3-fluoro-6-methoxybenzaldehyde (30 g, 129 mmol) in dichloromethane (200 mL) was added boron tribromide (65 g, 258 mmol) dropwise at -40°C, and the mixture was then stirred at 0°C for 3 hours. The reaction mixture was quenched at 0°C by adding methanol (40 mL) and saturated sodium bicarbonate solution (100 mL), and then extracted with ethyl acetate (×2). The organic layer was washed with saturated brine (×1), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography to give the target product (23 g, yield 80%) as a yellow solid.

[0183] 1H NMR (400MHz, CDCl3) δ11.78 (s, 1H), 10.35 (s, 1H), 7.32 (dd, J=7.6, 9.2Hz, 1H), 6.96 (dd, J=4.0, 9.2Hz, 1H).m / z=219[M+1] + .

[0184] Step D: 3-Bromo-2-((ethylamino)methyl)-4-fluorophenol

[0185]

[0186] To a solution of 2-bromo-3-fluoro-6-hydroxybenzaldehyde (20 g, 91.3 mmol) in methanol (100 mL) at 0°C was added a solution of ethylamine in tetrahydrofuran (2M, 91.3 mL, 182.6 mmol); stirred at 25°C for 30 minutes, and then sodium borohydride (6.9 g, 182.6 mmol) was added, and the reaction mixture was stirred at 25°C for 12 hours. The solvent was removed by vacuum concentration and the resulting mixture was diluted with water and extracted with ethyl acetate (×2). The organic layer was washed with brine (×1), dried over anhydrous sodium sulfate, filtered and concentrated to give the desired product as a white solid (15.9 g, yield 70%).

[0187] 1 H NMR (400MHz, CDCl3) δ6.93 (t, J=8.4Hz, 1H), 6.71 (dd, J=4.4, 8.4Hz, 1H), 4.23 (s, 2H), 2.76 (q, J=7.2Hz, 2H), 1.19 (t, J=7.2Hz, 3H).

[0188] Intermediate 3: tert-butyl 2-amino-5-chloro-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylate (Compound I3)

[0189]

[0190] Step A: (Z)-3-Amino-4,4,4-trichloro-2-cyano-butenoic acid ethyl ester

[0191]

[0192] At 0°C, triethylamine (2.0 g, 20 mmol) was added dropwise to an ethanol solution (120 mL) of ethyl cyanoacetate (41.2 g, 364 mmol) and trichloroacetonitrile (100 g, 693 mmol). The mixture was reacted at 0°C for 2 hours, and then the temperature was slowly raised to room temperature for 30 minutes. After the reaction was completed, the solvent was removed by concentration, and the residue was dissolved in dichloromethane and purified by chromatography on a silica gel column and eluted with dichloromethane to obtain the target product (93 g, yield 98%).

[0193] 1 H NMR (400MHz, CDCl3) δ10.20 (brs, 1H), 6.93 (brs, 1H), 4.30 (q, 2H), 1.33 (t, 3H).m / z=257[M+1] + .

[0194] Step B: 3,5-Diamino-1H-pyrazole-4-carboxylic acid ethyl ester

[0195]

[0196] At 0°C, hydrazine hydrate (80%, 58.4 g, 933 mmol) was slowly added dropwise to a solution of (Z)-3-amino-4,4,4-trichloro-2-cyano-butenoic acid ethyl ester (80 g, 311 mmol) in N,N-dimethylformamide (300 mL), and the reaction mixture was heated to 100°C and stirred for 1.5 hours. The solvent was concentrated to remove the residue, and the residue was slurried with dichloromethane and then allowed to stand overnight. The solid was collected by suction filtration, washed with dichloromethane, and dried to obtain the target product (35.5 g, 67% yield).

[0197] 1 H NMR (400MHz, DMSO-d6) δ10.4 (brs, 1H), 5.35 (brs, 4H), 4.13 (q, 2H), 1.24 (t, 3H).m / z=171[M+1] + .

[0198] Step C: Ethyl 2-amino-6-fluoro-5,7-dihydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate

[0199]

[0200] At room temperature, sodium methoxide (31.9 g, 590 mmol) and dimethyl 2-fluoromalonate (28.4 g, 189 mmol) were added to a solution of ethyl 3,5-diamino-1H-pyrazole-4-carboxylate (20 g, 118 mmol) in methanol (200 mL), and the reaction mixture was heated to 80°C for 5 hours. After the reaction was completed by TLC monitoring, the mixture was cooled to room temperature and concentrated under reduced pressure. Water was added to the residue, and the pH was adjusted to 3 with 2M hydrochloric acid aqueous solution to precipitate a large amount of white solid, which was filtered to obtain the title compound as a white solid (15.1 g, yield 50%).

[0201] m / z = 257 [M + 1] + .

[0202] Step D: Ethyl 2-amino-5,7-dichloro-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylate

[0203]

[0204] At room temperature, ethyl 2-amino-6-fluoro-5,7-dihydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate (15.1 g, 59 mmol) was added to phosphorus oxychloride (200 mL), and the reaction mixture was heated to 100°C for 12 hours. The mixture was cooled to room temperature and concentrated under reduced pressure, and an ice-water mixture was added to the residue, and the mixture was adjusted to weak alkalinity with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate (×2), and the ethyl acetate phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column and eluted with ethyl acetate / petroleum ether (1 / 2) to obtain the title compound (6.9 g, yield 40%).

[0205] m / z=293[M+1] + .

[0206] Step E: 2-amino-5-chloro-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester

[0207]

[0208] To a mixed solvent of 2-amino-5,7-dichloro-6-fluoropyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (6.9 g, 23.5 mmol) in ethanol (80 mL), tetrahydrofuran (30 mL) and water (60 mL) at 0°C, activated zinc powder (7.7 g, 117.5 mmol) and ammonium chloride (63 g, 117.5 mmol) were added; the mixture was stirred at 0°C for 0.5 hour, the reaction mixture was filtered, the filtrate was diluted with water, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to give the title compound (3.6 g, yield 60%).

[0209] m / z=259[M+1] + .

[0210] Intermediate 4: 2-amino-5-chloro-6-methoxypyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (Compound I4)

[0211]

[0212] Step A: (Z)-methyl 2,3-dimethoxyacrylate

[0213]

[0214] At 0°C, sodium hydride (60% purity, 112 g, 2.8 mol) was slowly added to a solution of methyl 2-methoxyacetate (208 g, 2.0 mol) and methyl formate (144 g, 2.4 mol) in tetrahydrofuran (2.0 L). During the addition, the temperature of the reaction system was kept below 0°C. After stirring at 0°C for 12 hours, a large amount of white solid was observed to be generated in the reaction system. Methyl tert-butyl ether was added to the reaction system, filtered, and the filter cake was vacuum dried to obtain the target crude product (350 g), which was directly used in the next step without purification.

[0215] Step B: Ethyl 2-amino-5-hydroxy-6-methoxypyrazolo[1,5-a]pyrimidine-3-carboxylate

[0216]

[0217] At room temperature, cesium carbonate (586 g, 1.8 mol) was added to a solution of (Z)-2,3-dimethoxyacrylate (263 g, 1.8 mol) and ethyl 3,5-diamino-1H-pyrazole-4-carboxylate (170 g, 1.0 mol) in N,N-dimethylformamide (2.0 L); the temperature was raised to 110° C. and stirred for 12 hours. After the reaction was completed as monitored by TLC, the reaction was cooled to room temperature, water was added to the reaction system to quench and dilute the reaction solution, and the pH value of the reaction solution was adjusted to about 6 with aqueous hydrochloric acid solution (5 M). A large amount of solid precipitated, which was filtered, and the filter cake was washed with methanol and dried in vacuo to obtain the target product (177 g, yield 70%).

[0218] m / z=253[M+1] + .

[0219] Step C: 2-amino-5-chloro-6-methoxypyrazole [1,5-a]pyrimidine-3-carboxylic acid ethyl ester

[0220]

[0221] At room temperature, ethyl 2-amino-5-hydroxy-6-methoxypyrazolo[1,5-a]pyrimidine-3-carboxylate (100 g, 0.4 mol) was added to phosphorus oxychloride (1.0 L). The reaction mixture was heated to 110° C. and stirred for 12 hours. It was concentrated under reduced pressure to remove most of the phosphorus oxychloride. Ice water was added to the residue, and the mixture was adjusted to weak alkalinity with saturated aqueous sodium carbonate solution. It was extracted with ethyl acetate (×3). The ethyl acetate phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to give the title compound (41 g, yield 39%).

[0222] m / z=271[M+1] + .

[0223] Intermediate 5: 2-amino-7-(benzyl(methyl)amino)-5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (Compound I5)

[0224]

[0225] Step A: Ethyl 2-amino-5,7-dihydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate

[0226]

[0227] At room temperature, sodium methoxide (54 g, 1.0 mol) and dimethyl malonate (53 g, 0.4 mol) were added to a solution of ethyl 3,5-diamino-1H-pyrazole-4-carboxylate (34 g, 0.2 mol) in methanol (200 mL), and the reaction mixture was heated to 80°C for 5 hours. After the reaction was completed by TLC monitoring, the mixture was cooled to room temperature and concentrated under reduced pressure. Water was added to the residue, and the pH was adjusted to 3 with 2M hydrochloric acid aqueous solution to precipitate a large amount of white solid, which was filtered to obtain the title compound as a white solid (29 g, yield 61%).

[0228] m / z=239[M+1] + .

[0229] Step B: Ethyl 2-amino-5,7-dichloropyrazolo[1,5-a]pyrimidine-3-carboxylate

[0230]

[0231] At room temperature, ethyl 2-amino-5,7-dihydroxypyrazolo[1,5-a]pyrimidine-3-carboxylate (10.7 g, 44.8 mmol) was added to phosphorus oxychloride (150 mL). The reaction mixture was heated to 110°C and stirred for 12 hours. It was concentrated under reduced pressure to remove most of the phosphorus oxychloride. Ice water was added to the residue, and the mixture was adjusted to weak alkalinity with saturated aqueous sodium carbonate solution. It was extracted with ethyl acetate (×3). The ethyl acetate phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to give the title compound (6.2 g, yield 50%).

[0232] m / z=275[M+1] + .

[0233] Step C: Ethyl 2-amino-5-chloro-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0234]

[0235] At room temperature, methylamine hydrochloride (7.4 g, 57.6 mmol) was added to a solution of ethyl 2-amino-5,7-dichloropyrazolo[1,5-a]pyrimidine-3-carboxylate (5.3 g, 19.2 mmol) and N,N-diisopropylethylamine (7.4 g, 57.6 mmol) in isopropanol (50 mL), and the reaction mixture was heated to 80°C for 5 hours. The reaction was monitored by TLC, cooled to room temperature, and concentrated under reduced pressure. Water was added to the residue, and extracted with ethyl acetate (×3). The ethyl acetate phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (4.7 g, yield 90%).

[0236] m / z = 270 [M + 1] + .

[0237] Step D: Ethyl 2-amino-7-(benzyl(methyl)amino)-5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate

[0238]

[0239] At room temperature, benzyl bromide (5.4 g, 31.4 mmol) was added to a solution of ethyl 2-amino-5-chloro-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylate (4.2 g, 15.7 mmol) and N,N-diisopropylethylamine (6.1 g, 47.1 mmol) in N,N-dimethylformamide (30 mL). The reaction was allowed to proceed at room temperature for 8 hours. After completion of the reaction as monitored by TLC, the mixture was cooled to room temperature and concentrated under reduced pressure. Water was added to the residue, and the mixture was extracted with ethyl acetate (×3). The ethyl acetate phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to give the title compound (5.3 g, yield 93%).

[0240] m / z=360[M+1] + .

[0241] Intermediate 6: 2-amino-6-chloroimidazole [1,2-b] pyridazine-3-carboxylic acid methyl ester (Compound I6)

[0242]

[0243] 2-Amino-6-chloroimidazole [1,2-b] pyridazine-3-carboxylic acid methyl ester (Compound I6) was synthesized according to the method of Example 45 intermediate in patent WO2017079519A1.

[0244] Intermediate 7: (R)-2-Hydroxypropyl pivalate (Compound I7)

[0245]

[0246] To a solution of (R)-1,2-propylene glycol (76 g, 1.0 mol) and triethylamine (122 g, 1.2 mol) in dichloromethane (1.0 L) was slowly added dropwise pivaloyl chloride (121 g, 1.0 mol) at 0°C. The reaction mixture was warmed to room temperature and stirred overnight. The reaction was completed after monitoring by TLC. The mixture was cooled to room temperature and concentrated under reduced pressure. Water was added to the residue, and the mixture was extracted with ethyl acetate (× 3). The ethyl acetate phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to give the title compound (128 g, yield 80%).

[0247] Intermediate 8: (S)-2-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)propan-1-amine (Compound I8)

[0248]

[0249] Step A: (S)-tert-Butyl (2-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)propyl)carbamate

[0250]

[0251] At 0°C, diisopropyl azodicarboxylate (8.1 g, 40 mmol) was slowly added to a solution of 3-bromo-2-((ethylamino)methyl)-4-fluorophenol (intermediate I2) (5 g, 20 mmol), triphenylphosphine (7.9 g, 30 mmol) and (R)-(2-hydroxypropyl)carbamic acid tert-butyl ester (5.3 g, 30 mmol) in tetrahydrofuran (60 mL); the reaction mixture was stirred at room temperature for 6 hours, and the reaction progress was monitored by TLC, and the reaction was basically completed. The reaction was cooled to room temperature, concentrated under reduced pressure, water was added to the residue, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (6.1 g, yield 75%).

[0252] m / z = 405 [M + 1] + .

[0253] Step B: (S)-2-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)propan-1-amine

[0254]

[0255] At 0°C, 4M HCl (19 mL) in 1,4-dioxane was slowly added dropwise to a solution of (S)-tert-butyl (2-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)propyl)carbamate (6.0 g, 14.8 mmol) in dichloromethane (50 mL). The resulting mixture was stirred at 30°C for 2 hours, and a large amount of white solid precipitated. The reaction progress was monitored by TLC, and the reaction was completed. The mixture was concentrated under reduced pressure to remove most of the solvent and hydrochloric acid gas, and water was added to the residue, and the mixture was diluted with saturated sodium carbonate aqueous solution to reduce the temperature. It was extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to give the title compound (4.5 g, yield 90%).

[0256] m / z = 305 [M + 1] + .

[0257] Intermediate 9: (R)-tert-butyl (3-hydroxybutyl)carbamate (Compound I9)

[0258]

[0259] (R)-tert-Butyl(3-hydroxybutyl)carbamate (Compound I9) was synthesized according to the intermediate synthesis method in Example 4 of patent WO2020001415A1.

[0260] Intermediate 10: (S)-3-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)butan-1-amine (Compound I10)

[0261]

[0262] The synthesis was carried out according to the synthesis method of intermediate 8 (compound I8), except that (R)-(2-hydroxypropyl)carbamic acid tert-butyl ester was replaced by (R)-(3-hydroxybutyl)carbamic acid tert-butyl ester.

[0263] Intermediate 11: (R)-2-(4-hydroxypentyl)isoindole-1,3-dione (Compound I11)

[0264]

[0265] Step A: 2-(4-oxopentyl)isoindole-1,3-dione

[0266]

[0267] At 0°C, sodium hydride (8.0 g, 60%, 0.2 mol) was slowly added in batches to a solution of phthalimide (29.4 g, 0.2 mol) in dry tetrahydrofuran (300 mL). After the addition was completed, the mixture was kept at 0°C and stirred for 0.5 hours. 5-chloro-2-pentanone (24.2 g, 0.2 mol) was added, and the mixture was slowly warmed to room temperature and stirred for 6 hours. The mixture was concentrated under reduced pressure to remove most of the solvent, diluted with water, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (40 g, yield 86%).

[0268] 1 H NMR (400MHz, CDCl3) δ7.85 (m, 2H), 7.72 (m, 2H), 3.72 (t, J=6.6Hz, 2H), 2.51 (t, J=7.0Hz, 2H), 2.15 (s, 3H), 1.97 (m, 2H).m / z=232[M+1] + .

[0269] Step B: 2-(4-Hydroxypentyl)isoindole-1,3-dione

[0270]

[0271] At room temperature, aluminum isopropoxide (88.4 g, 433 mmol) was slowly added in batches to a solution of 2-(4-oxopentyl)isoindoline-1,3-dione (40 g, 173 mmol) in isopropanol (300 mL), and the mixture was refluxed for 4 hours. Most of the solvent was removed under reduced pressure, 1 M hydrochloric acid was slowly added to neutralize, and the mixture was extracted with ethyl acetate (×3). The ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (20 g, yield 50%).

[0272] m / z=234[M+1] + .

[0273] Step C: (R)-5-(1,3-dioxoisoindol-2-yl)pentan-2-yl acetate

[0274]

[0275] At room temperature, 2-(4-hydroxypentyl)isoindole-1,3-dione (20 g, 85.7 mmol), vinyl acetate (44.3 g, 514 mmol) and lipase (8.2 g, 34.3 mmol) were added to isopropyl ether (500 mL), stirred at room temperature overnight, filtered through diatomaceous earth, concentrated to remove isopropyl ether, diluted with water, extracted with ethyl acetate (× 3), combined ethyl acetate phases, washed with saturated sodium chloride solution (× 1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to give the title compound (11 g, yield 47%).

[0276] m / z=276[M+1] + .

[0277] Step D: (R)-2-(4-Hydroxypentyl)isoindole-1,3-dione

[0278]

[0279] At 0°C, sodium methoxide (1.2 g, 21.8 mmol) was added to a solution of (R)-5-(1,3-dioxoisoindol-2-yl)pentane-2-yl acetate (5 g, 18.2 mmol) in methanol (50 mL), and the reaction was allowed to rise to room temperature for 3 hours. The reaction progress was monitored by TLC and the reaction was complete. The mixture was concentrated under reduced pressure to remove most of the solvent, diluted with water, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (3.8 g, yield 89%).

[0280] m / z=234[M+1] + .

[0281] Intermediate 12: (S)-4-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)pentan-1-amine (Compound I12)

[0282]

[0283] Step A: (S)-2-(4-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)pentyl)isoindole-1,3-dione

[0284]

[0285] The intermediate 8 (Compound I8) was synthesized according to the synthesis method of step A, except that (R)-(2-hydroxypropyl)carbamic acid tert-butyl ester was replaced by (R)-2-(4-hydroxypentyl)isoindole-1,3-dione.

[0286] m / z=463[M+1] + .

[0287] Step B: (S)-4-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)pentan-1-amine

[0288]

[0289] Methylamine alcohol solution (30 mL, 33 wt%) was added to a solution of (S)-2-(4-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)pentyl)isoindole-1,3-dione (5 g, 10.8 mmol) in methanol (50 mL) at room temperature, and the mixture was refluxed for 4 hours. The solvent was removed under reduced pressure, and the residue was separated by silica gel column chromatography to obtain the target product (2.9 g, yield 81%).

[0290] m / z = 333 [M + 1] + .

[0291] Intermediate 13: (S)-3-((1-aminopropan-2-yl)oxy)-2-((ethylamino)methyl)-6-fluorobenzonitrile (Compound I13)

[0292]

[0293] Step A: (S)-tert-butyl(2-(3-bromo-2-(((tert-butoxycarbonyl)(ethyl)amino)methyl)-4-fluorophenoxy)propyl)(tert-butoxycarbonyl)carbamate

[0294]

[0295] At room temperature, di-tert-butyl dicarbonate (21.5 g, 98.4 mmol) and 4-dimethylaminopyridine (1.0 g, 8.2 mmol) were added to (S)-2-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)propane-1-amine (5 g, 16.4 mmol) in dichloromethane (40 mL); the reaction was allowed to react for 6 hours at room temperature, and the reaction progress was monitored by TLC. The reaction was completed. The mixture was concentrated under reduced pressure to remove most of the solvent, diluted with water, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with a saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (9.2 g, yield 93%).

[0296] m / z = 605 [M + 1] + .

[0297] Step B: tert-Butyl (S)-(tert-Butyloxycarbonyl)(2-(2-(((tert-Butyloxycarbonyl)(ethyl)amino)methyl)-3-cyano-4-fluorophenoxy)propyl)carbamate

[0298]

[0299] To a degassed mixture of (S)-tert-butyl(2-(3-bromo-2-(((tert-butoxycarbonyl)(ethyl)amino)methyl)-4-fluorophenoxy)propyl)(tert-butoxycarbonyl)carbamate (9.0 g, 14.9 mmol), zinc cyanide (5.2 g, 44.7 mmol), zinc powder (195 mg, 3.0 mmol) and 1,1′-bis(diphenylphosphino)ferrocene (3.3 g, 6.0 mmol) in N,N-dimethylacetamide (100 mL) was added tris(dibenzylideneacetone)dipalladium (2.7 g, 3.0 mmol) at room temperature. The mixture was heated to 130° C. and stirred for 3 hours. The progress of the reaction was monitored by TLC and the reaction was complete. Filter through celite, wash with ethyl acetate (×3), wash with water (×1), wash with saturated sodium chloride solution (×1), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify on a silica gel column to obtain the title compound (6.5 g, yield 79%).

[0300] m / z=552[M+1] + .

[0301] Step C: (S)-3-((1-aminopropan-2-yl)oxy)-2-((ethylamino)methyl)-6-fluorobenzonitrile

[0302]

[0303] To a solution of tert-butyl (S)-(tert-butyloxycarbonyl)(2-(2-(((tert-butyloxycarbonyl)(ethyl)amino)methyl)-3-cyano-4-fluorophenoxy)propyl)carbamate (6.3 g, 11.4 mmol) in dichloromethane (50 mL) was slowly added dropwise 4M HCl (14 mL) in 1,4-dioxane at 0°C. The resulting mixture was stirred at 30°C for 2 hours, and a large amount of white solid precipitated. The reaction progress was monitored by TLC, and the reaction was completed. The mixture was concentrated under reduced pressure to remove most of the solvent and hydrochloric acid gas, and water was added to the residue, which was diluted with saturated aqueous sodium carbonate solution to reduce the temperature, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a silica gel column to give the title compound (2.7 g, yield 94%).

[0304] m / z = 252 [M + 1] + .

[0305] Intermediate 14: (S)-3-((4-amino-tert-butyl-2-yl)oxy)-2-((ethylamino)methyl)-6-fluorobenzonitrile (Compound I14)

[0306]

[0307] The product was synthesized according to the synthesis method of intermediate 13 (compound I13), except that (S)-2-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)propane-1-amine was replaced with (S)-3-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)butane-1-amine.

[0308] Intermediate 15: (S)-3-((5-aminopentan-2-yl)oxy)-2-((ethylamino)methyl)-6-fluorobenzonitrile (Compound I15)

[0309]

[0310] The product was synthesized according to the synthesis method of intermediate 13 (compound I13), except that (S)-2-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)propane-1-amine was replaced with (S)-4-(3-bromo-2-((ethylamino)methyl)-4-fluorophenoxy)pentane-1-amine.

[0311] Example 1 (S, 1 3 E, 1 4 E)-2-ethyl-4 5 -Fluoro-6-methyl-9-oxo-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0312]

[0313] Step A: Ethyl 5-((2-bromo-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0314]

[0315] At room temperature, 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (2.3 g, 10 mmol), 3-bromo-2-((ethylamino)methyl)-4-fluorophenol (2.5 g, 10 mmol), N,N-diisopropylethylamine (2.6 g, 20 mmol) and n-butanol (30 mL) were charged into the reactor. The resulting mixture was heated at 95°C for 12 hours. The reaction progress was monitored by TLC, and the reaction was substantially completed. The reaction was cooled to room temperature, concentrated under reduced pressure, water was added to the residue, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (3.8 g, yield 87%).

[0316] m / z=437[M+1] + .

[0317] Step B: (S)-ethyl 5-((2-bromo-3-fluoro-6-((1-pivaloyloxy)propan-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0318]

[0319] At 0°C, diisopropyl azodicarboxylate (1.4 g, 6.9 mmol) was slowly added to a solution of ethyl 5-((2-bromo-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate (2.0 g, 4.6 mmol), triphenylphosphine (1.8 g, 6.9 mmol) and (R)-2-hydroxypivalate propyl ester (0.9 g, 5.5 mmol) in tetrahydrofuran (20 mL); the reaction mixture was stirred at room temperature overnight, and the reaction progress was monitored by TLC. The reaction was basically completed. The reaction was cooled to room temperature and concentrated under reduced pressure. Water was added to the residue, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (1.8 g, yield 68%).

[0320] m / z=579[M+1] + .

[0321] Step C: (S)-5-((2-bromo-3-fluoro-6-((1-hydroxypropan-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0322]

[0323] To a solution of (S)-5-((2-bromo-3-fluoro-6-((1-pivaloyloxy)propane-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (1.8 g, 3.1 mmol) in ethanol (80 mL) and water (20 mL) at 0°C, lithium hydroxide monohydrate (2.6 g, 62.0 mmol) was slowly added. The resulting mixture was refluxed at 95°C overnight. The reaction was completed when the starting material disappeared as monitored by LCMS. The reaction was acidified with 4N HCl, extracted with ethyl acetate (×2), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a silica gel column to give the title compound (1.3 g, yield 90%).

[0324] m / z = 467 [M+1] +.

[0325] Step D: (S, 1 3 E, 1 4 E)-4 6 -Bromo-2-ethyl-4- 5 -Fluoro-6-methyl-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidin-4(1,2)-benzocyclononan-9-one

[0326]

[0327] At room temperature, (S)-5-((2-bromo-3-fluoro-6-((1-hydroxypropane-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (3.0 g, 6.4 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.84 g, 9.6 mmol), 4-dimethylaminopyridine (1.0 g, 8.4 mmol) and dichloromethane (200 mL) were charged into the reactor. The resulting mixture was refluxed at 40° C. overnight. The reaction was completed when the starting material disappeared as monitored by LCMS. The dichloromethane was removed in vacuo and the title compound (980 mg, 34% yield) was obtained by purification on a silica gel column.

[0328] 1H NMR (400MHz, DMSO-d6) δ8.65 (d, J=7.8Hz, 1H), 8.13 (s, 1H), 7.13-7.07 (m, 2H), 6.79 (d, J=7.9Hz, 1H), 5.68 (dd, J=14.2, 1.5Hz, 1H), 4.96-4.83 (m, 1H), 4.53 (dd, J=12.0, 3.6Hz, 1H), 4.16-4.05 (m, 2H), 3.86-3.77 (m, 2H), 1.46 (d, J=6.4Hz, 3H), 1.18 (t, J=7.0Hz, 3H).

[0329] m / z = 449 [M+1] +.

[0330] Step D: (S, 1 3 E, 1 4 E)-2-ethyl-4 5 -Fluoro-6-methyl-9-oxo-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0331]

[0332] At room temperature, the reactor was charged with (S, 1 3 E, 1 4 E)-4 6 -Bromo-2-ethyl-4- 5 -Fluoro-6-methyl-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-9-one (1.0 g, 2.2 mmol), cuprous cyanide (0.6 g, 6.6 mmol) and N,N-dimethylacetamide (10 mL). The resulting mixture was heated at 140°C under nitrogen protection for 8 hours. The reaction was completed after the disappearance of the starting material monitored by LCMS. The reaction mixture was cooled to room temperature, dichloromethane was added to the reaction mixture for dilution, and an appropriate amount of diatomaceous earth, an appropriate amount of activated carbon and an excess of buffer (H2O / NH4Cl / NH4OH=9.4 / 4.0 / 3.8) were added to the filtrate for filtration. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered through a one-inch diatomaceous earth bed and washed with DCM (×3). The layers were separated and the organic layer was washed with buffer (×2) and water (×2). The organic layer was concentrated to a minimum volume and purified by silica gel column to give the title compound (760 mg, yield 83%).

[0333] 1H NMR (400MHz, CDCl3) δ8.31 (dd, J=7.8, 4.5Hz, 1H), 8.23 ​​(d, J=10.4Hz, 1H), 7.08-6.97 ( m, 2H), 6.55 (dd, J=7.8, 3.7Hz, 1H), 5.91-5.81 (m, 1H), 5.45-5.36 (m, 0.4H), 4.85-4.7 6 (m, 0.6H), 4.71 (dd, J=11.8, 3.8Hz, 0.6H), 4.39 (dd, J=11.5, 5.4Hz, 0.4H), 4.34-4.2 3(m, 1H), 4.21-4.09(m, 1H), 3.99-3.80(m, 2H), 1.68-1.58(m, 3H), 1.31-1.24(m, 3H).

[0334] m / z=396[M+1] + .

[0335] Example 2 (S, 1 3 E, 1 4 E)-1 2 -amino-2-ethyl-4 5 -Fluoro-6-methyl-9-oxo-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0336]

[0337] Step A: tert-Butyl 2-amino-5-((2-bromo-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0338]

[0339] At room temperature, tert-butyl 2-amino-5-(p-toluenesulfonyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate (40.5 g, 100 mmol), 3-bromo-2-((ethylamino)methyl)-4-fluorophenol (29.8 g, 120 mmol), N,N-diisopropylethylamine (38.8 g, 300 mmol) and n-butanol (400 mL) were charged into the reactor. The resulting mixture was heated at 95°C for 12 hours. The reaction progress was monitored by TLC, and the reaction was substantially completed. The reaction was cooled to room temperature, concentrated under reduced pressure, water was added to the residue, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (33.6 g, yield 70%).

[0340] m / z=480[M+1] + .

[0341] Step B: tert-Butyl 2-amino-5-((2-cyano-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0342]

[0343] At room temperature, tert-butyl 2-amino-5-((2-bromo-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate (24 g, 50 mmol), cuprous cyanide (11.2 g, 125 mmol) and N,N-dimethylacetamide (120 mL) were charged into the reactor. The resulting mixture was heated at 100° C. for 96 hours. The progress of the reaction was monitored by TLC, and the reaction was substantially complete. The reaction was cooled to room temperature, dichloromethane was added to the reaction mixture for dilution, and an appropriate amount of diatomaceous earth, an appropriate amount of activated carbon and an excess of buffer (H2O / NH4Cl / NH4OH=9.4 / 4.0 / 3.8) were charged into the filtrate for filtration. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered through a one-inch diatomaceous earth bed and washed with DCM (×3). The layers were separated and the organic layer was washed with buffer (×2) and water (×2). The organic layer was concentrated to a minimum volume and purified by silica gel column to give the title compound (19.2 g, yield 90%).

[0344] m / z=427[M+1] + .

[0345] Step C: (S)-tert-butyl 2-amino-5-((2-cyano-3-fluoro-6-((1-pivaloyloxy)propan-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0346]

[0347] At 0°C, diisopropyl azodicarboxylate (2.0 g, 10.0 mmol) was slowly added to a solution of tert-butyl 2-amino-5-((2-cyano-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate (2.1 g, 5.0 mmol), triphenylphosphine (2.0 g, 7.5 mmol) and (R)-2-hydroxypivalate propyl ester (1.2 g, 7.5 mmol) in tetrahydrofuran (20 mL); the reaction mixture was stirred at room temperature for 6 hours, and the reaction progress was monitored by TLC, and the reaction was basically completed. The reaction was cooled to room temperature and concentrated under reduced pressure. Water was added to the residue, extracted with ethyl acetate (×3), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (2.3 g, yield 80%).

[0348] m / z=569[M+1] + .

[0349] Step D: (S)-2-amino-5-((2-cyano-3-fluoro-6-((1-hydroxypropan-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid hydrochloride

[0350]

[0351] To a solution of (S)-2-amino-5-((2-cyano-3-fluoro-6-((1-tert-valeryloxy)propane-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid tert-butyl ester (2.0 g, 3.5 mmol) in dichloromethane (20 mL) was slowly added dropwise 4M HCl (9 mL) in 1,4-dioxane at 0°C. The resulting mixture was stirred at 30°C for 2 hours, and a large amount of white solid precipitated. The reaction progress was monitored by TLC and the reaction was complete. The solid was filtered off, washed with MTBE (×2) and dried in a vacuum filter under nitrogen to obtain the title compound (1.7 g, yield 100%).

[0352] m / z=429[M+1] + .

[0353] Step E: (S, 1 3 E, 1 4 E)-1 2 -amino-2-ethyl-4 5 -Fluoro-6-methyl-9-oxo-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0354]

[0355] At room temperature, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.0 g, 5.6 mmol), 4-dimethylaminopyridine (86 mg, 0.7 mmol) and N,N-diisopropylethylamine (2.4 g, 18.5 mmol) were added to a solution of (S)-2-amino-5-((2-cyano-3-fluoro-6-((1-hydroxypropane-2-yl)oxy)benzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid hydrochloride (1.7 g, 3.7 mmol) in tetrahydrofuran (100 mL). The reaction mixture was heated to 80°C and stirred for 12 hours. The reaction progress was monitored by TLC and the reaction was completed. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. Water was added to the residue and extracted with ethyl acetate (×3). The ethyl acetate phases were combined and washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a silica gel column to give the title compound (760 mg, yield 50%).

[0356] 1 H NMR (400MHz, CDCl3) δ7.98 (d, J=7.5Hz, 1H), 7.05-6.88 (m, 2H), 6.30 (d, J=7.5Hz, 1H), 5.80 (d, J=14.6Hz, 1H), 4.96 (s, 2H), 4.77 (s, 1H), 4.5 8 (dd, J=11.5, 3.6Hz, 1H), 4.25 (dd, J=11.5, 2.8Hz, 1H), 4.08-4.01 (m, 1H), 3.77 (d, J=14.2Hz, 2H), 1.59 (d, J=6.3Hz, 3H), 1.36-1.16 (m, 3H).

[0357] m / z=411[M+1] + .

[0358] Example 3 (S, 1 3 E, 1 4 E, 9Z)-1 2 -amino-2-ethyl-4 5 -Fluoro-9-(hydroxyimino)-6-methyl-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0359]

[0360] At room temperature, the above Example 1 (410 mg, 1.0 mmol) was added to dry toluene (15 mL), and then phosphorus pentachloride (624 mg, 3.0 mmol) was added, heated to 80 ° C and stirred for 4 hours, cooled to room temperature, the solvent was spin-dried, and dry acetonitrile (15 mL) was added, cooled to 0 ° C, hydroxylamine hydrochloride (139 mg, 2.0 mmol), triethylamine (1.0 g, 10.0 mmol) were added, and the mixture was heated to room temperature and stirred for 4 hours. The reaction progress was monitored by TLC, and the reaction was completed. The reaction was cooled to room temperature, concentrated under reduced pressure, water was added to the residue, extracted with ethyl acetate (×4), the ethyl acetate phases were combined, washed with saturated sodium chloride solution (×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column to obtain the title compound (85 mg, yield 20%).

[0361] m / z=426[M+1] + .

[0362] Example 4 (S, 1 3 E, 1 4 E, 9Z)-1 2 -amino-2-ethyl-4 5 -fluoro-9-(methoxyimino)-6-methyl-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0363]

[0364] According to the synthesis method of Example 2, hydroxylamine hydrochloride was replaced by O-methylhydroxylamine hydrochloride to obtain (S, 1 3 E, 1 4 E, 9Z)-1 2 -amino-2-ethyl-4 5 -fluoro-9-(methoxyimino)-6-methyl-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0365] m / z=440[M+1] + .

[0366] Example 5 (S, 1 3 E, 1 4 E)-1 2 -amino-2-(ethyl-d5)-4 5 -Fluoro-6-methyl-9-oxo-5,8-dioxa-2-aza-1(5,3)-pyrazolo[1,5-a]pyrimidine-4(1,2)-benzocyclononane-4 6 -Formonitrile

[0367]

[0368] According to the synthesis method of Example 1, 3-bromo-2-((ethylamino)methyl)-4-fluorophenol was replaced with 3-bromo-2-(((ethyl-d5)amino)methyl)-4-fluorophenol to obtain the title compound.

[0369] m / z=416[M+1] + .

[0370] The following example compounds were synthesized by referring to the synthesis methods of the above intermediates and examples:

[0371]

[0372]

[0373]

[0374] Effect evaluation

[0375] 1. Compound enzyme inhibitory activity (IC 50 )Detection

[0376] Mobility shift assay was used to screen compounds for MET, SRC and CSF1R kinases. The core of this screening platform is MSA based on microfluidic chip technology, which applies the basic concept of capillary electrophoresis to the microfluidic environment. The experimental substrate is a fluorescently labeled polypeptide. Under the catalytic action of the enzyme in the reaction system, the substrate is converted into a product, and its charge also changes accordingly. MSA uses the difference in charge between the substrate and the product to separate the two and detect them separately.

[0377] The operation method is briefly described as follows:

[0378] Dissolve the compound powder in 100% DMSO and prepare a 10mM stock solution. The starting test concentration of the compound is 10,000nM, 3-fold gradient dilution, 10 concentrations, and duplicate well detection. Mix the gradient dilution compound and kinase in the Optiplate-384F well plate and incubate at room temperature for 10 minutes, then add ATP and Kinase substrate30 (GL Biochem, Cat.117885) mixture, mix and react at room temperature for 20 minutes. Add the stop detection solution to terminate the enzymatic reaction and read the conversion rate with Caliper EZ Reader II.

[0379] Data Analysis:

[0380]

[0381] Where: Conversion%_sample is the conversion rate reading of the sample; Conversion%_min: the mean of the negative control wells, representing the conversion rate reading of the wells without enzyme activity; Conversion%_max: the mean of the positive control well ratio, representing the conversion rate reading of the wells without compound inhibition.

[0382] Fitting the dose-effect curve: With the log value of the concentration as the X-axis and the percentage inhibition rate as the Y-axis, the log(inhibitor) vs.response-Variable slope of the analysis software GraphPad Prism 5 was used to fit the dose-effect curve, thereby obtaining the IC value of each compound for the inhibition of enzyme activity. 50 value.

[0383] Table 1. c-Met, SRC and CSF1R kinase inhibitory activities of compounds

[0384]

[0385] a: TPX-0022 has the following structure, and its synthesis can refer to Example 5 of WO2019023417A1

[0386]

[0387] b: Comparative compound 42 has the following structure, and its synthesis refers to Example 1 of WO2019206069A1

[0388]

[0389] 2. Compound Cell Activity (IC 50 )Detection

[0390] 2.1. Cellular activity against human non-small cell lung cancer cells HCC827 (IC 50 )Detection

[0391] Human non-small cell lung cancer cells HCC827 cells were cultured in an incubator (37°C, 5% CO2) with 1640 medium plus 10% FBS (fetal bovine serum) and 1% P / S (penicillin / streptomycin). In the detection of the compound, HCC827 cells were plated in a 96-well transparent flat-bottom black wall plate (Corning, Cat#3603) at a concentration of 3000 cells / 90 μL per well. The compound was diluted 3 times from 10 mM to 11 concentrations to prepare a 10-fold drug solution, and then 10 μL was added to 90 μL of cell culture medium (DMSO final concentration was 0.1%, v / v). After 72 hours of treatment, 100 μL of CellTiter-Glo (Promega, Cat#G7572) was added and the cells were shaken on an orbital shaker for 5 minutes to lyse the cells. The cell plate was then placed at room temperature for 20 minutes to stabilize the cold light signal. The luminescence value was read using a SpectraMax multi-label microplate reader (MD, 2104-0010A).

[0392] Data Analysis:

[0393] The data were analyzed using GraphPad Prism 5.0 software, and the dose-effect curves were fitted using nonlinear S-curve regression, from which the IC50 values ​​were calculated.

[0394] Cell survival rate (%) = (Lum 待测药 -Lum 培养液对照 ) / (Lum 细胞对照 -Lum 培养液对照 )×100%.

[0395] The test results show that Example 1 has a very significant inhibitory effect on HCC827 cells.

[0396] 2.2. Cell activity (IC 50 )Detection

[0397] The culture medium of human gastric cancer cell line SNU-5 is IMDM basal medium supplemented with 10% FBS (fetal bovine serum) and 1% P / S (penicillin / streptomycin), and the culture medium of Ba / F3-TEL-CSF1R stable transfection cell line is RPMI-1640 medium supplemented with 10% FBS and 1% P / S. Both cell lines were cultured in a carbon dioxide incubator at 37°C and a CO2 concentration of 5%.

[0398] In the detection of compounds, SNU-5 and Ba / F3-TEL-CSF1R cells were plated in 96-well cell culture plates (Corning, Cat#3610) at a concentration of 3000 cells / 100 μL per well and cultured overnight. The compound was diluted 3 times with DMSO starting from 200 μM, and a total of 9 concentrations were diluted to prepare a 200-fold drug solution. 3 μl of the 200-fold concentration compound was diluted with 197 μl of complete culture medium to obtain a 3-fold concentration compound. 50 μl of the latter was added to the cell well plate and cultured for 72 hours. Before the test, the cell culture plate was equilibrated to room temperature, 40 μL of Cell Titer-Glo (Promega, Cat#G7571) was added to each well, and the cells were lysed by shaking for two minutes. The cell plate was then placed at room temperature for 60 minutes to stabilize the cold light signal. The luminescence value was read using a Perkin Elemer Envision multi-function plate reader.

[0399] Data Analysis:

[0400] Luminescence values ​​are processed using the following formula:

[0401] %Inh=(Max signal-Compound signal) / (Max signal-Min signal)x 100

[0402] Max signal: value of DMSO-treated group; Min signal: value of blank culture medium group.

[0403] The processed data were analyzed by GraphPad Prism 5.0 software, and the dose-effect curve was obtained by fitting the data using nonlinear S-curve regression, and the IC 50 value.

[0404] The test results show that the example compounds have a very significant inhibitory effect on SNU-5 and Baf3-TEL CSF1R cells; see Table 2 for specific data.

[0405] Table 2. Inhibitory activity of compounds on cells

[0406]

[0407] 3. Synergistic killing of tumor cells with AZD9291

[0408] Human lung cancer cells H1975 cells (L858R and T790M double mutations) and HCC827 cells were cultured in an incubator (37°C, 5% CO2) with 1640 medium plus 10% FBS (fetal bovine serum) and 1% P / S (penicillin / streptomycin). In the detection of the compound, H1975 cells or HCC827 were plated in a 96-well plate (Corning) at a concentration of 3000 cells / 195 μL per well, and the compound was diluted 3-fold to 11 concentrations starting from 10 mM. 4 μL of each concentration was added to 96 μL of 1640 medium to dilute to 25× compound, and then 5 μL was added to 195 μL of cell culture medium (DMSO final concentration was 0.1%, v / v), and 35 μL of (purchased from Promega), the fluorescence signal was measured on Flex Station 3 (Molecular Devices) according to the operating procedures in the instruction manual, and the IC of the compound on cell proliferation inhibition was calculated using GraphPad Prism 5.0 50 The Chou-Talalay combination index method was used to calculate the combined drug effect. The combination index (CoI) value of 0.9≤CI≤1.1 was additive, 0.8≤CI<0.9 was low synergistic, 0.6≤CI<0.8 was moderate synergistic, 0.4≤CI<0.6 was high synergistic, and 0.2≤CI<0.4 was strong synergistic.

[0409] The experimental results show that the combination of Example 1 and AZD9291 has moderate to high synergistic effects on EGFR double mutant cells H1975 (L858R and T790M double mutations) and HCC827 cells (MET overexpression), indicating that the combination of Example 1 and EGFR inhibitors can overcome EGFR resistance.

[0410] 4. Pharmacokinetic Experiments

[0411] Male SD rats were divided into groups, 3 in each group, and the example compound (1 mg / kg) was administered orally by single oral gavage and intravenous injection. The animals were fasted overnight before the experiment, and the fasting time was from 10 hours before administration to 4 hours after administration. Blood was collected at 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration in the oral group, and at 0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 hours after injection in the intravenous group. 0.3 mL of whole blood was collected through the fundus venous plexus after isoflurane anesthesia using a small animal anesthesia machine, placed in a heparin anticoagulation tube, and the sample was centrifuged at 4°C and 4000rpm for 5 minutes. The plasma was transferred to a centrifuge tube and stored at -80°C until analysis. The sample in the plasma was extracted using protein precipitation, and the extract was analyzed by LC / MS / MS.

[0412] Embodiment 1 and embodiment 2 have good pharmacokinetic and pharmacodynamic properties.

[0413] 5. Blood-brain distribution experiment

[0414] Male SD rats were divided into groups, 12 rats in each group, and the example compound (10 mg / kg) was orally administered by a single oral gavage. The animals were fasted overnight before the experiment, and the fasting period was from 10 hours before administration to 4 hours after administration. According to the previous pharmacokinetic data, the rats in the Example 1 group were killed 0.25, 0.5, 2 and 6 hours after administration; the rats in the TPX-0022 group were killed 0.25, 1, 4 and 8 hours after administration, and blood and brain tissue were collected. After the samples were processed, they were centrifuged at 4°C and 4000rpm for 5 minutes, and the plasma was transferred to a centrifuge tube and stored at -80°C until analysis. The samples in the plasma were extracted using the protein precipitation method, and the extract was analyzed by LC / MS / MS. The results show that the Example 1 compound can enter the brain tissue, and the drug brain / blood distribution ratio is 1.16-6.4, which is far higher than TPX-0022. The brain / blood distribution of the Example 1 compound and TPX-0022 is as follows Figure 1 and Figure 2 shown.

[0415] Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the spirit and scope of the invention as defined in the accompanying claims.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is a compound represented by the following formula:

2. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 2, in the preparation of a medicament for treating a tyrosine kinase-mediated disease.

4. The use according to claim 3, wherein The tyrosine kinase is selected from one or more of the following: SRC, MET, CSF1R, ALK, ROS1, TRKA, TRKB, TRKC, JAK2, SRC, FYN, LYN, YES, FGR, FAK, AXL, ARK5.

5. The use according to claim 3, wherein The tyrosine kinase-mediated disease is selected from the group consisting of cancer, pain, neurological disease, autoimmune disease and inflammation.

6. A combination drug for treating cancer in a patient, comprising a therapeutically effective amount of a preparation for inhibiting SRC and MET and / or CSF1R and an additional anticancer agent, administered simultaneously or separately in the same or different strengths, wherein: The preparation for inhibiting SRC, MET and / or CSF1R comprises the compound according to claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 2; the additional anticancer agent is an EGFR antibody or an EGFR small molecule inhibitor.

7. The combined drug according to claim 6, wherein: The EGFR antibody or EGFR small molecule inhibitor is selected from cetuximab, nacitozumab, panitumumab or amivantamab dual antibody, afatinib, brigatinib, canertinib, dacomitinib, erlotinib, gefitinib, HKI 357, lapatinib, osimertinib, naquatinib, nazartinib, neratinib, omotinib, pelitinib, PF-06747775, rociletinib, vandetanib, ametinib, vometinib, Mobocertinib, DZD9008, BEBT-109, lazertinib, CLN-081, WTS-004, JFAN-1001, C-005, XZP-5809-TT1, JRF103, FWD1509, JNJ-372, or a pharmaceutically acceptable salt thereof.

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