Benzamide compound and application thereof in degradation of DDRs

Abstract

The invention belongs to the field of biological medicines, and particularly relates to a benzamide compound or a pharmaceutically acceptable salt, a stereoisomer, a solvate or a polymorphic substance thereof. According to the present invention, the downstream key protein is regulated by degrading or inhibiting the discoidin domain receptor 1 (DDR1) so as to further achieve the effect of treating the related diseases; the invention creates a compound which is novel in structure, remarkable in biological activity and capable of effectively inhibiting or degrading DDR1, and the compound can be used for treating diseases related to DDR1 homeostasis imbalance.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to a benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug for degrading receptor tyrosine kinase, especially discoidin domain receptors (DDRs), and further discloses its application. Background Art

[0002] Discoidin domain receptors (DDR) are a member of the receptor tyrosine kinase (RTKs) family and play an important role in the signal transduction pathway that controls cell proliferation and differentiation. Studies have shown that DDR signaling disorders can lead to a variety of abnormal states of cells and play an important role in the development and regulation of carcinoid cells. Diseases associated with DDR dysfunction include cancer, nervous system degeneration, chronic inflammatory diseases, and fibrosis. Therefore, the study and practical application of DDR regulation methods have opened up a new way to prevent and treat the occurrence of these diseases in clinical practice. At present, the DDR receptor family includes two members, DDR1 and DDR2, of which DDR1 includes five subtypes: DDR1a, DDR1b, DDR1c, DDR1d, and DDR1e. Among them, DDR1 is mainly expressed in epithelial cells, smooth muscle cells, fibroblasts, oligodendrocytes, and macrophages in the lung, kidney, colon, and brain, while DDR2 is mainly expressed in fibroblasts, myofibroblasts, smooth muscle cells, and chondrocytes in the kidney, skin, lung, heart, and connective tissue.

[0003] DDR1, a collagen receptor, is composed of three main domains, namely the extracellular ligand binding region, the transmembrane region, and the intracellular tyrosine kinase region. Among them, the extracellular domain is composed of a discoid domain and a quasi-diskoid domain that can bind to the ligand collagen; the transmembrane domain includes an extracellular juxtamembrane domain and a transmembrane helical region, which can mediate collagen-independent receptor dimerization (Yeung DA, J mol Biol. 2019; 431: 368-390); the intracellular domain includes an intracellular juxtamembrane domain and a catalytic tyrosine kinase domain. In particular, DDR1 has an activation mechanism different from other RTKs, which require collagen binding stimulation to activate downstream pathways, leading to autophosphorylation. DDR1 preferentially binds to collagens IV and VIII, and has a lower affinity for type X collagen. Studies have shown that the function of DDR1 is mainly achieved through three pathways, namely the classical collagen-DDR1 signaling pathway, which depends on DDR1 kinase activity and directly affects downstream molecules such as Shc, Nck2, and Shp-2; or the non-classical DDR1 signaling pathway, which depends on collagen binding but is independent of DDR1 kinase activity; or, it can exert its effects independently of collagen stimulation and DDR1 kinase activity (Valiathan RR, et al. Cancer Metastasis Rev. 2012; 31(1-2): 295-321).

[0004] Studies have shown that DDR1 is expressed in a variety of tumors, including lung cancer, breast cancer, brain tumors, ovarian cancer, esophageal cancer, head and neck tumors, liver cancer, testicular cancer, etc., and its high expression is closely related to poor tumor prognosis (Rammal H, Front Pharmacol. 2016; 7: 55). For example, DDR1 expression in cancer cell tissues of patients with non-small cell lung cancer is positively correlated with mortality, and DDR1 expression and phosphorylation levels in lung cancer tissues are significantly increased. According to immunohistochemical analysis of 171 cases of non-small cell lung cancer, the positive rate of DDR1 in aggressive non-small cell lung cancer is as high as 61% (Yang SH. Oncol Rep. 2010; 24: 311-319). At the same time, DDR1 can promote the proliferation and growth of a variety of malignant tumor cells. In human colon cancer and breast cancer, DDR1 can upregulate the anti-apoptotic protein Bcl-xL through the activation of downstream signaling proteins Ras / Raf / ERK and PI3K / Akt pathways, thereby allowing cancer cells to survive under toxic stress (Matadag SP. Medicinal Chemistry Research, 2021, 30(3): 535-551). High expression of DDR1 increases resistance to chemotherapy in breast cancer and lymphoma cell lines and mediates signals that promote cell survival.

[0005] In addition, DDR1 activated by natural type IV collagen can induce increased expression of CD9 in cancer cells in the breast cancer MDA-MB-231 cell line, leading to cancer cell migration; it also induces the secretion and invasion of matrix metalloproteinase-2 and metalloproteinase-9 through DDR1 and Src-dependent pathways, increasing their cellular expression levels. DDR1 activation plays an important role in the invasive ability and metabolic activity of breast cancer cells. Similarly, the upregulation of these two metalloproteinases caused by DDR1 is also a necessary condition for the metastasis and invasion of liver cancer cells (Lee, JH. Sci. Rep. 2018, 8, 1). In addition, in vivo and in vitro experiments have shown that DDRs are involved in the differentiation of tumor cells; further, they also promote cell malignant transformation and tumor invasion and metastasis by destroying normal cell-matrix signaling, which can help tumors establish barriers around them to prevent T cells from infiltrating and killing tumor cells. Studies have shown that DDR1 can increase the invasion and migration ability of non-small cell lung cancer cells by promoting epithelial-mesenchymal cell transformation; DDR1 can also regulate T cells, CD4 + and CD8 + To inhibit anti-tumor immune response, thereby promoting the growth of breast cancer (Zhong, X. Oncol Rep. 2019; 42(6): 2844-2854).

[0006] Clinical trials have shown that the combination of anti-tumor immunotherapy and targeted tumor therapy can significantly improve the survival rate of patients with advanced tumors. Immunotherapy targeting anti-cytotoxic T lymphocyte-associated antigen (CTLA)-4 and anti-programmed death receptor 1 (PD)-1 / PD-ligand (L) 1 plays an important role in the treatment of melanoma, colorectal cancer and breast cancer. According to the infiltration distribution of cytotoxic immune cells in the tumor microenvironment (TME), tumors can be divided into immune-inflammatory tumors and immune-rejection or immune-desert tumors. Immunoinflammatory tumors are usually accompanied by high T cell infiltration, PD-L1 expression and tumor mutation load, which makes tumor cells respond to treatment. In contrast, CD8 +The number of T cells is low. In between are tumors with altered phenotypes, which are further divided into two different patterns: "rejection" and "immunosuppression". In some cases, T cells appear at the edge of the tumor (infiltrating edge) but cannot infiltrate. This "excluded" phenotype reflects the intrinsic ability of the host immune system to effectively initiate T cell-mediated immune responses, and the ability of tumors to evade such responses by physically hindering T cell infiltration. In other cases, tumor sites show low levels of immune infiltration, indicating the lack of physical barriers and the presence of an immunosuppressive environment that limits further recruitment and expansion. This can be defined as an "immunosuppressive" phenotype. Therefore, targeting immune-rejecting tumors and using drugs to eliminate or inhibit these factors that are not conducive to the function of immune checkpoint inhibitors will help improve the clinical immunotherapy effect of tumors. DDR1 can weaken the clearance of cancer cells by the immune system through its effects on the tumor microenvironment. DDR1 binds to collagen in the extracellular matrix (ECM) through its extracellular domain (ECD), densely arranging collagen fibers and presenting a highly ordered state, hindering the infiltration of immune cells and weakening their ability to kill tumor cells. Studies have shown that in a mouse model with DDR1 knockout, the infiltration of T cells in the tumor improved and tumor growth was inhibited. It can be inferred that DDR1 gene knockout or inhibition of DDR1 protein is a potential method to block the tumor's anti-immune surveillance ability. DDR1 may become a new target for tumor immunotherapy (Sun X, Nature, 2021, 599(7886):673-678). Preclinical studies have shown that DDR1 inhibitors exhibit a wide range of anti-tumor activity in tumor transplantation mouse models derived from patients with non-small cell lung cancer, gastric cancer, esophageal cancer, liver cancer, breast cancer, and colorectal cancer. Studies have shown that anti-DDR1 monoclonal antibodies can inhibit the immune rejection of solid tumors. In view of the formation of physical barriers and immune rejection of DDR1 in the tumor microenvironment, regulation of this protein is expected to be an effective means to improve existing tumor immunotherapy.

[0007] In addition to its role in tumor pathogenesis, DDRs are also key mediators of the secretion of a variety of inflammatory cytokines, and are dysregulated in a variety of inflammatory diseases including osteoarthritis, organ fibrosis, and atherosclerosis (Leitinger, B. Int. Rev. Cellmol. Biol. 2014, 310, 39-87). DDR1 is also associated with fibrotic lesions in the liver, kidneys, lung tissue, and blood vessels. Studies have shown that DDR1 not only directly stimulates the secretion of inflammatory factors, but also enhances the effects of other proinflammatory cytokines and bacterial products (Matsuyama, WJ Immunol. 2004, 172, 2332–2340). DDR1 activation is associated with a variety of kidney diseases. DDR1 activation is involved in the pathology of a variety of kidney diseases, including Alport syndrome caused by type IV collagen gene mutations, IgA nephropathy, Goodpasture syndrome, lupus nephritis, ANCA (anti-neutrophil cytoplasmic antibodies)-related nephritis, and diabetic nephropathy. In various renal disease models, inhibition of DDR expression can prevent inflammatory changes and fibrosis in the kidney. In a model of anti-glomerular basement membrane glomerulonephritis with prominent inflammation and unilateral ureteral obstruction, it was found that inhibition of DDR1 expression can reduce inflammatory cell migration and hinder fibrosis formation (Guerrot, D. Am. J. Pathol. 2011, 179, 83).

[0008] It can be seen that DDR1 is dysregulated in a variety of inflammatory diseases such as atherosclerosis, osteoarthritis, and organ fibrosis. Collagen-induced activation of DDR1b can significantly promote the production of interleukin 8 (IL-8), macrophage inflammatory protein 1α (MIP-1α), and monocyte chemoattractant protein 1 (MCP-1) by macrophages during inflammatory responses. DDR1 knockout mice can reduce lipopolysaccharide (LPS)-induced chemokine secretion and reduce mortality. Small molecule DDR1 inhibitors can reduce the release of inflammatory cytokines and have a significant therapeutic effect in mouse inflammation models. Experiments have confirmed that DDR1 deficiency can effectively reduce bleomycin-induced lung inflammation and pulmonary fibrosis (Avivi-Green, C.Am.J.Respir.Crit.Care Med.2006,174,420–427). In addition, DDR1 is also associated with pathological conditions such as systemic lupus erythematosus, osteoarthritis, heparin-induced thrombocytopenia, and vitiligo vulgaris.

[0009] Given that DDR1 is closely associated with cancer, inflammation, fibrosis, and neurodegenerative diseases, it is expected to become an important target for intervention and treatment of these diseases. Regulation of DDR1 can be achieved through monoclonal antibodies that act on its extracellular binding region or small molecule inhibitors that act on the intracellular kinase domain and block downstream signaling pathways. Among them, small molecule inhibitors can be divided into multi-target RTK inhibitors and selective DDR1 inhibitors. Because the structures of the kinase ATP binding regions of some RTKs family members are highly homologous, especially between DDRs and c-Kit and Bcr-Abl kinases, many RTK multi-target kinase inhibitors have certain DDR1 inhibitory activity. For example, the multi-target inhibitors Nilotinib (Nilotinib, I), Imatinib (Imatinib, II), and Dasatinib (Dasatinib, III) can inhibit DDR1 autophosphorylation induced by collagen. At present, a variety of Bcr-Abl inhibitors that have been marketed, such as Ponatinib, Bosutinib, and Bafetinib, show strong DDR1 inhibitory activity. In addition, the p38MAPK inhibitor Doramapimod, the B-Raf / VGFR dual-target inhibitor Sorafenib, and the c-Met / VEGFR-2 dual-target inhibitor Foretinib kinase inhibitors have strong binding affinity to the DDR1 kinase domain.

[0010]

[0011] In addition, some DDR1 inhibitors with certain selectivity have also been discovered, and some compounds have potential application prospects in the treatment of tumors, neurodegenerative diseases, Alzheimer's disease, dry eye syndrome, inflammatory and fibrotic diseases (William AD Biomolecules. 2021, 11 (11): 1671). DDR1 small molecule inhibitors include urea, quinazoline, isoquinazoline, tetrahydroisoquinazoline, 1H-indole, spirocyclic indoline and triheterospirocyclic decanone (WO2017 / 038870, WO2016064970A1, WO2015060373A1, CN108276388A, WO2017137334A1, WO2017 / 038871).

[0012] Benzylamide compound (DDR-IN-1, IV) is a DDR1 inhibitor with certain selectivity that evolved from the structure of small molecule inhibitor of kinase Bcr-Abl, and can inhibit DDR1 autophosphorylation in cells at nanomolar concentration (Kim, HG.; et al. ACS Chem. Biol. 2013, 8, 2145). Further research has derived small molecule inhibitors with DDR1 as the main target but with different kinase spectra. Compound (7rh, V) shows significant selectivity for DDR1 among 456 kinases, and relatively weak inhibitory activity against DDR2, Bcr-Abl and c-Kit (Gao, M. et al. J. Med. Chem. 2013, 56, 3281, CN101885722B). The compound (7ae, VII) produced by structural optimization of the above compounds exhibited excellent DDR1 affinity and had certain anti-inflammatory activity against LPS-induced acute lung injury in mice (Wang, Z.; et al. J. Med. Chem. 2016, 59, 5911; Wang, Z.; et al. ACS Med. Chem. Lett. 2017, 8, 327). Similarly, compound (7f, VI) significantly reduced the inhibitory effect on TrkA, TrkB and TrkC while retaining the ability to inhibit DDR1. The compound showed a certain therapeutic effect in a tumor mouse model (Zhu, D.; et al. J. Med. Chem. 2019, 62, 7431).

[0013]

[0014] Although the biological activity of these DDRs inhibitors has been confirmed by many experiments, like most other RTK inhibitors, this type of small molecule drugs relies on the competition for the ATP binding domain, and long-term use can cause cell resistance. At the same time, DDR has both kinase catalytic function and non-catalytic function of other domains. Simply relying on the competition of small molecule inhibitors cannot offset all its functions. Therefore, it is urgent to find a more effective means to regulate DDR1.

[0015] At present, the method of using the ubiquitin-protease system (UPS) in eukaryotic cells to regulate intracellular protein levels has received great attention in the past decade. UPS negatively regulates signaling functions by inducing protein degradation efficiently and specifically, and it can degrade 80%-90% of ubiquitinated proteins in cells. It plays an extremely important role in maintaining the levels of various proteins in cells, involving almost all life activities such as regulating cell cycle, proliferation, apoptosis, metastasis, gene expression, and signal transduction. Ubiquitin protein, composed of 76 amino acids, is a highly conserved protein that is widely present in eukaryotic cells. The ubiquitination mark of protein initiates the degradation of substrate proteins, and this process is carried out under the synergistic action of ubiquitin activating enzyme E1, ubiquitin conjugating enzyme E2, and E3 ubiquitin ligase. First, ubiquitin is connected to E1 through a high-energy thioester bond between the carboxyl group on its C-terminal glycine and the essential cysteine ​​thiol group on the ubiquitin activating enzyme E1, becoming an activated ubiquitin; secondly, the activated ubiquitin is transferred from the ubiquitin activating enzyme E1 to the ubiquitin conjugating enzyme E2; finally, under the action of the E3 ubiquitin ligase, the ubiquitin molecule connected to the ubiquitin conjugating enzyme E2 is connected to the substrate protein through a covalent connection of an isopeptide bond. After being ubiquitinated, the substrate protein can be transported to the 26S proteasome or enter the lysosome for digestion and degradation. The specific recognition ability of the E3 ubiquitin ligase for substrate proteins determines the specificity of ubiquitin-mediated protein degradation.

[0016] Proteolysis targeting chimeras (PROTACs) are a class of bifunctional molecules that use the intracellular ubiquitin-proteasome system to degrade specific proteins. This technology will be able to bind to small molecule ligands of specific targeted proteins, connect them with E3 ubiquitin ligase ligands through bridge fragments, and form a bifunctional compound. Through the optimized link position and the length of the bridge chain, the ligands at both ends of the PROTAC molecule bind to the target protein and the E3 ubiquitin ligase respectively, forming a target protein-PROTACs-E3 ligase ternary complex, thereby promoting the ubiquitination tagging of the target protein, and then being degraded by the proteasome system. Due to the characteristics of ternary complex formation, the target protein ligand used in PROTAC technology does not need to have strong target binding activity, so it can target target proteins that traditional inhibitors cannot drug, such as skeleton proteins and transcription factors; in addition, the overall degradation of the target protein helps to overcome the drug resistance problem of small molecule inhibitors; PROTAC molecules act repeatedly in a catalytic mechanism, and PROTAC-induced degradation is event-driven rather than occupancy-driven. After the complex is formed and ubiquitin transfer is completed, the drug dissociates and transfers to the next target in an enzymatic manner. To a certain extent, it is possible to maintain effective drug activity at low doses. The technical difficulty of PROTAC lies in the relationship between target protein degradation and generation rate, the ability of target protein to bind ubiquitin, the conformation and site of the connection between target protein ligand and E3 ubiquitin ligase ligand, the modification of the length and composition of the bridge chain, and the concentration affect the formation and stability of the ternary complex, so it is more challenging to regulate. Among the more than 600 known E3 ubiquitin ligases, only a limited number of them are actually used in the design of PROTAC compounds, including CRBN, VHL, MDM2, and cIAP1. These E3 ubiquitin ligases confer substrate specificity to achieve ubiquitination of target proteins. Among the common E3 ubiquitin ligases, the Von Hippel-Lindau (VHL) tumor suppressor is composed of extensins B and C, Cul2 and Rbx1, and its main substrate is hypoxia-inducible factor 1 (HIF-1). Studies on the ligands of the E3 ubiquitin ligase VHL have obtained the crystal structure of its complex, thus confirming that small molecule compounds can mimic the main substrate of the transcription factor HIF-1 (CN108601764A). During the application process, the binding of the ligand of VHL to the E3 ubiquitin ligase is relatively weak, which easily causes incomplete degradation of the target protein and causes off-target effects. Another important E3 ligase, Cereblon (CRBN), is a protein encoded by the human CRBN gene, and the CRBN homologous gene is highly conserved, showing its importance in physiology.Cereblon forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB1), Cullin-4A (CUL4A) and Cullin-1 regulator (ROC1). The complex can ubiquitinate a series of proteins. Considering the binding ability of the phthalimide structural unit to CRBN, the structural unit is often used as an E3 ligase recruitment ligand to hijack CRBN to degrade the target protein. So far, there are many kinds of experimental drugs developed using PROTAC technology, including nuclear protein receptor degraders, kinase degraders, transcription factor degraders, etc. Small molecule inhibitors and PROTACs targeting DDR1 can block kinase matrix signal transduction, prevent inactivated kinases from transmitting oncogene signals through their skeleton functions, and lead to continuous functional loss of DDR1. The present invention designs and develops inhibitors or degraders targeting DDR1 with unique structures. Such compounds can treat related diseases by regulating DDR1, and therefore have great clinical significance. Summary of the invention

[0017] The first technical problem to be solved by the present invention is to provide a benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope label or prodrug, which inhibits and / or degrades discoidin domain receptors (DDRs), regulates downstream key proteins and further plays a role in treating related diseases, and shows outstanding ability to inhibit tumor cell proliferation;

[0018] The second technical problem to be solved by the present invention is to provide a preparation method and application of the benzamide compounds.

[0019] In order to solve the above technical problems, a benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug described in the present invention has a structure shown in the following formula (I):

[0020]

[0021] in,

[0022] The R a Selected from H, C1-C3 straight chain or branched alkyl, or C1-C3 straight chain or branched alkyl in which some or all of the hydrogen atoms are replaced by fluorine;

[0023] The R b Selected from H or C1-C3 straight or branched alkyl;

[0024] The G is selected from CR1 or N;

[0025] The R cand R1 are independently selected from H, halogen, R2, -OR2, -(CH2) x NR3R4, -(CH2) y CONR3R4 or a 5-7 membered aromatic heterocycle containing at least one nitrogen atom; wherein R2 is selected from C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; wherein R3 or R4 are independently selected from H, C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; or, R3 and R4 and the nitrogen atom to which they are connected together form a 3-7 membered azacycloalkyl group containing 1-2 nitrogen atoms, and the hydrogen atoms on the nitrogen can be replaced by C1-C3 straight or branched alkyl; wherein x and y are independently selected from integers of 0-2;

[0026] Ar is selected from aromatic rings or wherein A and B are independently selected from N or CH, and at least one of them is N; wherein the asterisk (*) indicates the connection site with the M group, and the wavy line indicates the connection site with the alkynyl carbon;

[0027] The M is selected from H, R5, -NHR5 or a functional group consisting of two parts EL-**, wherein the R5 is selected from a C1-C3 straight chain or branched alkyl group; the E is a ligand part capable of binding to the E3 binding enzyme cereblon, which is connected to the bridge part L through a covalent bond; wherein the double asterisk (**) represents the asterisk connection site with the Ar group.

[0028] As a preferred solution, the R c and R1 are independently selected from H, NH2, CF3, OCH3, N(CH3)2, CONH2, CH2N(CH3)2 or

[0029] Specifically, the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, wherein E has the structure shown in E1 or E2 below:

[0030] or

[0031] Wherein, the D1 is selected from CH2 or C=O, preferably CH2; the D2 is selected from CH or N; wherein, the (#) position indicates the position where E1 or E2 is covalently bonded to the L part, and the connection position is at position 1 or 2 of the E1 or E2 structural formula.

[0032] Specifically, in the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, E is selected from at least one of the following structures:

[0033]

[0034] Specifically, in the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope label or prodrug, the L part is a bridge group connecting the E part and the Ar part; the L is selected from the L1-L7 structures shown below, wherein the wavy line indicates the position where L is covalently bonded to the asterisk position of the Ar part; the asterisk (*) indicates the position where L is covalently bonded to the pound position (#) of the E part;

[0035]

[0036] Wherein, m is selected from a natural number between 0 and 2, n is selected from a natural number between 0 and 1, and p is selected from a natural number between 1 and 2;

[0037] Said T1 is selected from O, NH or CONH;

[0038] Said T2 is selected from O or NH;

[0039] Said T3 is selected from N or CH;

[0040] The T4 is selected from NH or absent;

[0041] The T5 is selected from O or CONH;

[0042] The T6 is selected from O or NH;

[0043] Said T7 is selected from CH2 or CO or does not exist;

[0044] The T8 is selected from CO or does not exist;

[0045] The Cy is selected from a 4-7 membered azetidinyl group, preferably an azetidinyl group containing one nitrogen atom, preferably azetidine or azetidinyl group.

[0046] Specifically, in the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, L is selected from at least one of the following structures:

[0047]

[0048] Specifically, the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, the compound is selected from one of the following structures:

[0049]

[0050]

[0051] The present invention also provides a benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug having a structure shown in the following formula (II):

[0052]

[0053] in,

[0054] The R d is selected from H, C1-C3 straight chain or branched chain alkyl, or C1-C3 straight chain or branched chain alkyl in which some or all of the hydrogen atoms are replaced by fluorine;

[0055] The R e Selected from H or C1-C3 straight or branched alkyl;

[0056] The G' is selected from CR6 or N;

[0057] The R f , R6 are independently selected from H, halogen, R7, -OR7, -(CH2) x NR8R9, -(CH2) y CONR8R9 or a 5-7 membered aromatic heterocycle containing at least one nitrogen atom; wherein R7 is selected from C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; wherein R8 or R9 are independently selected from H, C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; or, R8 and R9 and the nitrogen atom to which they are connected together form a 3-7 membered azacycloalkyl group containing 1-2 nitrogen atoms, and the hydrogen atoms on the nitrogen can be replaced by C1-C3 straight or branched alkyl; wherein x and y are independently selected from integers of 0-2;

[0058] Ar' is selected from aromatic rings or wherein A and B are independently selected from N or CH, and at least one of them is N; wherein the asterisk (*) indicates the connection site with the M group, and the wavy line indicates the connection site with the alkynyl carbon;

[0059] The M' is selected from a C1-C3 straight or branched alkyl group or a functional group consisting of two parts E'-L'-**, wherein the E' is a ligand part capable of binding to the E3 binding enzyme cereblon, which is connected to the bridge part L' through a covalent bond; wherein the double asterisk (**) represents the asterisk connection site with the Ar' group.

[0060] As a preferred solution, the R f , R6 are independently selected from H, NH2, CF3, OCH3, N(CH3)2, CONH2, CH2N(CH3)2 or

[0061] Specifically, the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug:

[0062] When Ar' is When M' is composed of two parts: EL-**; or,

[0063] When Ar' is When M' is a C1-3 straight chain or branched chain alkyl or is composed of two parts EL-**.

[0064] Specifically, the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, wherein E' has the structure shown in E1' or E2' as follows:

[0065] or

[0066] Wherein, D1 is selected from CH2 or C=O, preferably CH2; D2 is selected from CH or N;

[0067] In the formula, the (#) position indicates the position where E1' or E2' is covalently bonded to the L' part, and the connection position is at position 1 or 2 of the E1' or E2' structural formula.

[0068] Specifically, in the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, the E' is selected from at least one of the following structures:

[0069]

[0070] Specifically, in the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, the L' portion is a bridge group connecting the E' portion and the Ar portion;

[0071] The L' is selected from the structure L1'-L7' shown below, wherein the wavy line indicates the position where L' is covalently bonded to the asterisk position of the Ar' part; the asterisk (*) indicates the position where L' is covalently bonded to the pound position (#) of the E part;

[0072]

[0073] Wherein, m is selected from a natural number between 0 and 2, n is selected from a natural number between 0 and 1, and p is selected from a natural number between 1 and 2;

[0074] Said T1 is selected from O, NH or CONH;

[0075] Said T2 is selected from O or NH;

[0076] Said T3 is selected from N or CH;

[0077] The T4 is selected from NH or absent;

[0078] The T5 is selected from O or CONH;

[0079] The T6 is selected from O or NH;

[0080] Said T7 is selected from CH2 or CO or does not exist;

[0081] The T8 is selected from CO or does not exist;

[0082] The Cy is selected from a 4-7 membered azetidinyl group, preferably an azetidinyl group containing one nitrogen atom, preferably azetidine or azetidinyl group.

[0083] Specifically, in the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, the L' is selected from at least one of the following structures:

[0084]

[0085] Specifically, the benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, the compound is selected from one of the following structures:

[0086]

[0087]

[0088] The present invention also discloses a pharmaceutical composition, comprising the benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug, and a pharmaceutically acceptable carrier or excipient.

[0089] Specifically, the pharmaceutical composition further comprises at least one additional bioactive agent;

[0090] Preferably, the bioactive agent comprises at least one of an anticancer agent, an immunomodulator, an immune checkpoint inhibitor, a kinase inhibitor, a corticosteroid or an anti-inflammatory agent.

[0091] The present invention also discloses the use of the compound or the pharmaceutical composition in preparing a pharmaceutical preparation for degrading or inhibiting discoidin domain receptor 1 (DDR1).

[0092] Specifically, the discoidin domain receptor 1 (DDR1) includes a wild type and a structural mutant type.

[0093] The present invention also discloses the use of the compound or the pharmaceutical composition for preparing a drug for preventing, diagnosing or treating a disease related to discoidin domain receptor 1 (DDR1).

[0094] Specifically, the discoidin domain receptor 1 (DDR1)-related diseases or conditions include cancers, immune-related diseases (including inflammatory diseases), fibrotic diseases, and neurodegenerative diseases associated with abnormal expression of DDR1.

[0095] Specifically, the cancer includes gastric cancer, intestinal cancer, esophageal cancer, head and neck cancer, lung cancer, liver cancer, brain cancer, breast cancer, colorectal cancer, skin cancer, thyroid cancer, prostate cancer, soft tissue cancer, endometrial cancer, uterine cancer, testicular cancer, cervical cancer, ovarian cancer, fallopian tube tumors, leukemia, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, renal cell carcinoma, bladder cancer, kidney cancer, pancreatic cancer, lymphoma, non-Hodgkin's lymphoma, melanoma, myeloproliferative disease, sarcoma, angiosarcoma, peripheral neuroepithelioma, glioma, astrocytoma, oligodendroglioma, ependymoma, glioblastoma, neuroblastoma, gangliocytoma, ganglioglioma, medulloblastoma, pineal cell tumor, meningioma, meningiosarcoma, neurofibroma or neurothecoma.

[0096] Specifically, the immune-related diseases (including inflammatory diseases) include rheumatoid arthritis, autoimmune encephalomyelitis, ankylosing spondylitis, axial spondyloarthritis, psoriasis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, recurrent oral ulcers, Kawasaki disease, spondyloarthritis, neuromyelitis optica, Behcet's disease, lupus nephritis, familial Mediterranean fever, ulcerative colitis, autoimmune hepatitis, asthma, and arteriosclerosis.

[0097] Specifically, the fibrotic disease includes pulmonary fibrosis, liver fibrosis, renal fibrosis or Crohn's disease.

[0098] Specifically, the neurodegenerative diseases include Alzheimer's disease, amyotrophic lateral sclerosis, ataxia telangiectasia, bovine spongiform encephalopathy, Creutzfeldt-Jakob disease, Huntington's disease, cerebellar atrophy, multiple sclerosis, Parkinson's disease, primary lateral sclerosis, spinal muscular atrophy, cerebral ischemia, spastic paraplegia or myasthenia gravis.

[0099] The pharmaceutical preparations involved in various uses of the present invention can be selected from preparations suitable for oral administration, injection administration, and inhalation administration. These dosage forms contain the active ingredients described in the invention to prepare the desired pharmaceutical preparations. In the therapeutic use of the pharmaceutical preparations, the daily dosage of the compound is in accordance with the conventional dosage and can be varied according to the needs of the patient, the severity of the disease to be treated, and the compound used.

[0100] The present invention provides a benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof. The compound regulates downstream key proteins by degrading and / or inhibiting discoidin domain receptor 1 (DDR1) and further plays a role in treating related diseases. The present invention creates a compound with novel structure, significant biological activity, and the ability to effectively inhibit or degrade DDR1, and the compound can be used to treat diseases related to DDR1 homeostasis imbalance.

[0101] The present invention creatively designs and develops a benzylamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof with a novel and unique structure. Such compounds can degrade or inhibit discoidin domain receptor 1 and regulate related proteins to further play a role in diagnosing, preventing and treating a variety of diseases, which is of great significance for the development of new drugs.

[0102] The present invention creatively designs and develops a benzylamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof with a novel and unique structure, which effectively improves the solubility performance of the compound and has better pharmaceutical performance. BRIEF DESCRIPTION OF THE DRAWINGS

[0103] In order to make the content of the present invention more clearly understood, the present invention is further described in detail below according to specific embodiments of the present invention in conjunction with the accompanying drawings, wherein:

[0104] Figure 1 The results of the degradation and phosphorylation inhibition of DDR1 by the representative compounds of the present invention are shown;

[0105] Figure 2 The results are the verification results of the mechanism of degradation of DDR1 by the representative compounds of the present invention through the ubiquitin-proteasome system;

[0106] Figure 3 The dendrogram is a result of measuring the inhibition rate of representative compounds of the present invention on 76 tyrosine kinases. DETAILED DESCRIPTION

[0107] The present invention is further described below in conjunction with the examples and accompanying drawings of the examples thereof. These examples are only used for more detailed specific descriptions and should not be construed as limiting the present invention in any form. The present invention can be implemented in a variety of different ways defined and covered by the claims. In this application, unless otherwise stated, the use of "or" or "and" means "and / or". In addition, the use of the term "including" and other forms, such as "including", "containing" and "having", is not restrictive. The section headings used herein are only for organizational purposes and should not be interpreted as limitations on the subject matter described.

[0108] Although many materials and methods of operation used in the present invention are well known in the art, the present invention is still described as detailed as possible. In the following, if not specifically stated, the materials and methods of operation used are well known in the art. Unless otherwise defined, all terms (including technical terms and scientific terms) used in the present invention have the same meaning as those of ordinary skill in the art to which the present invention belongs. If not specifically stated, the starting materials used in the synthesis method to prepare the compounds of the present invention are obtained from commercial channels or can be prepared according to known methods reported in the literature in the art or described herein.

[0109] The following examples of the present invention relate to the separation, purification and structural identification of synthetic compounds, wherein the medium-pressure preparation instrument used for the separation of the synthetic compounds and intermediates in each example is Isolera Prime produced by Biotage; the preparative high performance liquid chromatography uses a Gilson GX-281 instrument, and the reversed-phase silica gel column comes from Agela Technologies; the high performance liquid chromatography purification conditions are: separation column type XBridge-1, filler particle size 5 μm, diameter 13 mm, length 150 mm; the ultraviolet detector is set at 254 or 220 nm; eluent: 0.1% TFA aqueous solution and solvent methanol or acetonitrile; the flow rate is 20 mL / min; the LC-MS uses Agilent Technologies 1260 Infinity II; wherein the chromatographic column is a Symmetry C18 column, the filler particle size is 5 μm, the diameter is 4.6 mm, and the length is 50 mm; the mobile phase uses 0.05% formic acid aqueous solution and 0.05% formic acid acetonitrile solution; the mass spectrometry ionization source is electrospray; and the mass spectrometry acquisition is Agilent Technologies 6120; Waters Q-Tof instrument was used for high-resolution mass spectrometry, and the precise molecular weight of the compound was determined within the range of ±10ppm of the calculated value; Bruker Ascend 400 was used for nuclear magnetic resonance spectroscopy; tetramethylsilane and deuterated DMSO were used as internal standards; the unit part per million (ppm) was represented by the symbol δ.

[0110] Example 1

[0111] In this example, the compound 4-methyl-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]-N-phenylbenzamide (Compound A1) was synthesized.

[0112]

[0113] Synthesis of Compound A1-1

[0114] Compound A1-SM1 (500 mg, 2.36 mmol), trimethylethynylsilane (695 mg, 7.07 mmol), CuI (90 mg, 0.47 mmol), Pd(PPh3)2Cl2 (331 mg, 0.47 mol), DIEA (1219 mg, 9.43 mmol) and DMF (5 mL) were added to a 50 mL single-mouth bottle in sequence. The reaction was carried out at 100 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (10 mL) and extracted five times with EA (50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A1-1 (700 mg, brown oil, purity 70.727%) was obtained. Yield: 91.54%; LCMS (ESI) m / z calcd.for C 12 H 15 N3Si[M+H] + 230.36 found 230.3. It can be seen that the product structure is correct.

[0115] Synthesis of Compound A1-2

[0116] Compound A1-1 (1 g, 0.0044 mol), TBAF solution (5 mL, 10 mol / L) and THF (5 mL) were added to a 50 mL single-mouth bottle in sequence. The reaction was carried out at 25 °C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (10 mL) and extracted three times with EA (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography with an eluent and a ratio of PE / EtOAc = 20 / 1 to 5 / 1 to obtain compound A1-2 (300 mg, yellow solid, purity 97.031%). Yield: 43.18%; LCMS (ESI) m / z calcd.for C9H7N3[M+H] + 158.06; found 158.3; 1 H NMR (400 MHz, DMSO-d6): δ = 9.32 (d, J = 1.7 Hz, 1H), 8.49 (d, J = 1.9 Hz, 1H), 6.59 (s, 1H), 4.46 (s, 1H), 2.43 (s, 3H). It can be seen that the product structure is correct.

[0117] Synthesis of Compound A1-5

[0118] Compound A1-SM2 (2 g, 0.1025 mmol), LiOH.H2O (0.91 g, 0.205 mmol) and THF / MeOH / H2O=2 / 1 / 1 (6 mL) were added to a 25 mL single-mouth bottle. The mixture was reacted at 25°C for 16 hours under nitrogen protection. After the reaction, the reaction solution was adjusted to pH=4 with dilute hydrochloric acid, and the precipitated solid was filtered and dried to obtain compound A1-5 (1.5 g, white solid, purity 87.777%). Yield: 69.44%; LCMS (ESI) m / z calcd. for C8H7IO2[M+H] + 262.95found261.0; 1 HNMR (400MHz, DMSO_d6): δ=8.31(s,1H),7.85(d,J=7.8Hz,1H),7.45(d,J=7.9Hz,1H),2.43(s,3H). It can be seen that the product structure is correct.

[0119] Synthesis of Compound A1-6

[0120] Compound A1-5 (200 mg, 0.76 mmol), aniline (90 mg, 1.53 mmol), HATU (280 mg, 1.53 mmol), DIEA (395 mg, 3.05 mmol) and DMF (20 mL) were added to a 50 mL single-mouth bottle in sequence. The reaction was carried out at 25 ° C for 0.5 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (10 mL) and extracted three times with EA (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was separated by a preparative plate (PE / EA=2 / 1) to obtain compound A1-6 (200 mg, white solid, purity 96.930%). Yield: 75.34%; LCMS (ESI) m / zcalcd.for C 14 H 12 INO[M+H] + 338.0;found338.1; 1 H NMR (400MHz, DMSO_d6): δ=10.25(s,1H),8.40(d,J=1.5Hz,1H),7.91(dd,J=7.9,1.5Hz,1H),7.76(d,J=8.0Hz,2H),7.48(d,J=7.9Hz,1H),7.35(t,J=7.8Hz,2H),7.11(t,J=7.4Hz,1H),2.45(s,3H). It can be seen that the product structure is correct.

[0121] Synthesis of Compound A1

[0122] Compound A1-6 (50 mg, 0.148 mmol), A1-2 (47 mg, 0.297 mmol), CuI (5.7 mg, 0.0297 mmol), Pd(PPh3)2Cl2 (21 mg, 0.0297 mol), DIEA (76 mg, 0.593 mmol) and DMF (5 mL) were added to a 50 mL single-mouth bottle in sequence. The mixture was reacted at 40 °C for 16 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (5 mL) to precipitate a solid. The solid was slurried with EtOAc and DCM to obtain a filter cake. The filter cake was dissolved with DMSO and then dripped into water to precipitate a solid. Compound A1 (11.9 mg, brown solid, purity 96.037%) was obtained by filtration. Yield: 21.04%; LCMS (ESI) m / z calcd.for C 23 H 18 N4O[M+H] + 367.15; found 367.2; 1 H NMR (400MHz, DMSO-d6): δ=10.30(s,1H),9.44(s,1H),8.65(s,1H),8.17(s,1H),7.93(d,J=7.5Hz,1H),7.79(d,J=7.5Hz,2H),7.52(d,J=7.9Hz,1H),7.36(t,J=7.2Hz,2H),7.11(s,1H),6.64(s,1H),2.58(s,3H),2.46(s,3H). It can be seen that the product structure is correct.

[0123] Example 2

[0124] In this example, the compound 4-ethyl-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]-N-phenylbenzamide (Compound A2) was synthesized.

[0125]

[0126] Synthesis of Compound A2-2

[0127] Compound A2-1 (100 mg, 0.362 mmol), aniline (67 mg, 0.724 mmol), HATU (275 mg, 0.724 mmol), DIEA (187 mg, 1.449 mmol) and DMF (1 mL) were added to a 50 mL single-mouth bottle in sequence. The reaction was carried out at 25 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (5 mL) and extracted three times with EA (15 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was separated by a preparative plate (PE / EA=2 / 1) to obtain compound A2-2 (50 mg, white solid, purity 98.6%). Yield: 38.76%; LCMS (ESI) m / zcalcd.for C 15 H 14 INO[M+H] + 352.01;found352.1; 1 H NMR (400MHz, DMSO_d6): δ=10.25(s,1H),8.39(d,J=1.4Hz,1H),7.93(dd,J=7.9,1.4Hz,1H),7.76(d,J=8.1Hz,2H),7.46(d,J=8.0Hz,1H),7.35(t,J=7.8Hz,2H),7.11(t,J=7.3Hz,1H),2.75(q,J=7.5Hz,2H),1.18(t,J=7.5Hz,3H). It can be seen that the product structure is correct.

[0128] Synthesis of Compound A2

[0129] Compound A2-2 (50 mg, 0.142 mmol), A1-2 (45 mg, 0.285 mmol), CuI (5.4 mg, 0.0285 mmol), Pd(PPh3)2Cl2 (20 mg, 0.0284 mol), DIEA (74 mg, 0.570 mmol) and DMF (1 mL) were added to a 50 mL single-necked bottle in sequence, and the mixture was reacted at 40°C for 16 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (5 mL) to precipitate a solid. The solid was slurried with EtOAc and DCM to obtain a filter cake. The filter cake was dissolved with DMSO and then dropped into water to precipitate a solid. The solid was separated by high pressure preparation (column type: XBridge-1, 5 μm, 19-150 mm; detector: 254 nm; mobile phase: acetonitrile / water (0.1% formic acid solution); elution gradient: 35-70 / 8 minutes) to obtain compound A2 (18.98 mg, yellow solid, purity 98.765%). Yield: 34.62%; LCMS (ESI) m / z calcd.for C 24H 20 N4O[M+H] + 381.16; found 381.3; 1 H NMR (400MHz, DMSO_d6): δ=10.31(s,1H),9.43(d,J=1.9Hz,1H),8.63(d,J=2.0Hz,1H),8.16(d,J=1.5Hz,1H),7.96(dd,J=8.0,1.6Hz,1H),7.79(d,J=8.1Hz,2H),7.53(d,J=8.1Hz,1H),7.36(t,J=7.8Hz,2H),7.11(t,J=7.3Hz,1H),6.64(s,1H),2.95(q,J=7.5Hz,2H),2.46(s,3H),1.29(t,J=7.5Hz,3H). It can be seen that the product structure is correct.

[0130] Example 3

[0131] In this example, the compound N-(3-aminophenyl)-4-ethyl-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]benzamide (Compound A3) was synthesized.

[0132]

[0133] Synthesis of Compound A3-1

[0134] At room temperature, A3-SM1 (550 mg, 1.896 mmol) was dissolved in 10 mL DMF, and A1-2 (360 mg, 2.282 mmol) was added, followed by Pd(PPh3)2Cl2 (132 mg, 0.189 mmol), CuI (36 mg, 0.189 mmol), and TEA (955 mg, 9.445 mmol). The argon gas was replaced three times, and the temperature was raised to 40 ° C for 18 h. The reaction progress was monitored by TLC (developing solvent: PE: EA = 4: 1). After the reaction was completed, the reaction was stopped and the solvent was evaporated. Saturated NaCl and ethyl acetate (30 mL × 3) were used for extraction, the ethyl acetate phases were combined, the organic phases were dried and concentrated, and purified by column chromatography (PE: EA = 50: 1 to 5: 1) to obtain compound A3-1 (500 mg, yellow solid). Yield: 82.58%; 1H NMR (500MHz, DMSO-d6) δ9.56 (s, 1H), 8.69 (s, 1H), 8.38–8.30 (m, 1H), 8.10 (d, J = 6.8 Hz, 1H), 7.58–7.48 (m, 1H), 6.83 (s, 1H), 3.87 (d, J = 8.8 Hz, 3H), 3.00–2.89 (m, 2H), 2.46 (s, 3H), 1.27 (d, J = 7.5 Hz, 3H). It can be seen that the product structure is correct.

[0135] Synthesis of Compound A3-2

[0136] Under magnetic stirring, in a 100mL single-mouth bottle, A3-1 (250mg, 0.783mmol) was dissolved in MeOH (10mL), and the solution was light yellow and turbid. LiOH aqueous solution (10mL) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for reaction. TLC was used to monitor the reaction progress (developing solvent: PE:EA=4:1), and the reaction was stopped after 16h. After the reaction was completed, the reaction solution was concentrated to remove most of the methanol and water, and 1M HCl was used to adjust the pH to 4. A large amount of white solid precipitated, which was filtered and dried. Compound A3-2 (200mg, white solid) was obtained. Yield: 83.68%. The compound was used directly in the next step.

[0137] Synthesis of Compound A3

[0138] A3-2 (100 mg, 0.327 mmol) was dissolved in DCM (5 mL), and m-phenylenediamine (39 mg, 0.361 mmol), DIEA (127 mg, 0.981 mmol), and PyBop (190 mg, 0.361 mmol) were added. After stirring for 5 min, the mixture was moved to a 30°C oil bath for reaction. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 30:1), and the reaction was stopped after 16 h. Water was added to the reaction solution, extracted with EA (20 ml x 3), dried over anhydrous Na2SO4, and the organic phase was concentrated and purified by agar plate (DCM: MeOH = 50:1) for 3 times. Compound A3 (22 mg, yellow solid, purity 86.5%) was obtained. Yield: 14.7%; LCMS (ESI) m / z calcd. For C 24 H 21 N5O[M+H] + 396.17; found 396.31; 1HNMR(500MHz,DMSO-d6)δ10.01(s,1H),9.57(dd,J=2.1,0.9Hz,1H),8.71(d,J= 2.1Hz,1H),8.34(d,J=2.3Hz,1H),8.13(d,J=1.9Hz,1H),7.50(d,J=8.1Hz,1H), 7.13–7.08(m,1H),6.96(t,J=7.9Hz,1H),6.89–6.82(m,2H),6.31(ddd,J=7.9,2 .2,1.0Hz,1H),5.08(s,2H),3.01–2.91(m,2H),2.51(s,3H),1.31–1.26(m,3H). It can be seen that the product structure is correct.

[0139] Example 4

[0140] In this example, compound A4 was synthesized using N-[5-amino-3-(trifluoromethyl)phenyl]-4-ethyl-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]benzamide.

[0141]

[0142] Synthesis of Compound A4

[0143] A3-2 (100 mg, 0.328 mmol) and A4-SM (87 mg, 0.492 mmol) were dissolved in DCM (5 mL), DIEA (127 mg, 0.984 mmol) and PyBop (214 mg, 0.412 mmol) were added, and the mixture was stirred for 5 min and then moved to a 30°C oil bath. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 30:1). The reaction was stopped after 24 h, and the raw acid and amine were not completely reacted. Water was added to the reaction solution, extracted with DCM (20 ml x 3), dried over anhydrous Na2SO4, and the organic phase was concentrated and prepared by plate preparation to obtain compound A4 (33 mg white solid, purity 95.21%). Yield: 15.76%; LCMS (ESI) m / z calcd. For C 25 H 20 F3N5O[M+H] + 464.16;found464.1; 1H NMR (500MHz, DMSO-d6) δ10.29 (s, 1H), 9.58 (d, J = 2.0 Hz, 1H), 8.71 (d, J = 2.1 Hz, 1H), 8.35 (d, J = 2.3 Hz, 1H), 8.17 (d, J = 2.0 Hz, 1H), 7.53 (d, J = 8.1 Hz, 1H), 7.40–7.36 (m, 1H), 7.28 (d, J = 2.0 Hz, 1H), 6.85 (d, J = 2.3 Hz, 1H), 6.61 (d, J = 1.9 Hz, 1H), 5.65 (d, J = 8.4 Hz, 2H), 3.00–2.92 (m, 2H), 2.52 (s, 3H), 1.33–1.27 (m, 3H). It can be seen that the product structure is correct.

[0144] Example 5

[0145] In this example, the compound N-(3-methoxyphenyl)-4-ethyl-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]benzamide (Compound A5) was synthesized.

[0146]

[0147] Synthesis of Compound A5

[0148] A3-2 (100 mg, 0.327 mmol) was dissolved in DMF (2 mL), TEA (100 mg, 0.983 mmol) and HATU (136 mg, 0.359 mmol) were added, and after stirring for 5 min, A5-SM (48 mg, 0.393 mmol) dissolved in DMF (1 mL) was added, and the mixture was moved to a 30°C oil bath. The reaction progress was monitored by TLC (developing solvent: PE:EA=2:1), and the reaction was stopped after 20 h. After the reaction was completed, water was added to the reaction solution, and the mixture was extracted with EA (20 mlx3). The organic phase was concentrated after drying over anhydrous Na2SO4, and purified by preparative plate to obtain compound A5 (33 mg, yellow solid, purity 95.2%). Yield: 23.37%; LCMS (ESI) m / z calcd.for C 25 H 22 N4O2[M+H] + 411.17;found411.2; 1H NMR (500MHz, DMSO-d6) δ10.29 (s, 1H), 9.58 (s, 1H), 8.71 (s, 1H), 8.39–8.32 (m, 1H), 8.17 (s, 1H), 7.53 (d, J=8.1Hz, 1H), 7.48 (s, 1H), 7.39 (d, J=8.0Hz, 1H), 7.32–7.23 (m, 1H), 6.85 (s, 1H), 6.73–6.67 (m, 1H), 3.76 (s, 3H), 3.00–2.92 (m, 2H), 2.46 (s, 3H), 1.34–1.27 (m, 3H). It can be seen that the product structure is correct.

[0149] Example 6

[0150] In this example, the compound N-(3-trifluoromethylphenyl)-4-ethyl-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]benzamide (Compound A6) was synthesized.

[0151]

[0152] Synthesis of Compound A6

[0153] A3-2 (100 mg, 0.327 mmol) was dissolved in DMF (5 mL), TEA (100 mg, 0.983 mmol) and HATU (136 mg, 0.359 mmol) were added, and after stirring for 5 min, A6-SM (64 mg, 0.393 mmol) dissolved in DMF (1 mL) was added, and the mixture was moved to a 30°C oil bath. The reaction progress was monitored by TLC (developing solvent: PE:EA=2:1), and the reaction was stopped after 28 h. The raw acid and amine were not completely reacted. Water was added to the reaction solution, extracted with EA (20 mlx3), dried over anhydrous Na2SO4, and the organic phase was concentrated and purified by preparative plate to obtain compound A6 (32 mg, white solid, purity 94.71%). Yield: 21.79%; LCMS (ESI) m / z calcd.for C 25 H 19 F3N4O[M+H] + 449.15;found449.2; 1H NMR(500MHz,DMSO-d6)δ10.62(s,1H),9.58(dd,J=2.1,0.9Hz,1H),8.71(d,J=2 .1Hz,1H),8.35(d,J=2.3Hz,1H),8.26(d,J=2.0Hz,1H),8.21(d,J=1.9Hz,1H),8 .11–8.06(m,1H),7.65–7.58(m,1H),7.56(d,J=8.0Hz,1H),7.50–7.44(m,1H), 6.85(dd,J=2.3,0.9Hz,1H),3.01–2.93(m,2H),2.46(s,3H),1.36–1.27(m,3H). It can be seen that the product structure is correct.

[0154] Example 7

[0155] In this example, the compound N-[3-(dimethylamino)phenyl]-4-ethyl-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]benzamide (Compound A7) was synthesized.

[0156]

[0157] Synthesis of Compound A7

[0158] A3-2 (100 mg, 0.328 mmol) was dissolved in DMF (4 mL), TEA (100 mg, 0.983 mmol) and HATU (125 mg, 0.328 mmol) were added, and after stirring for 5 min, A7-SM (58 mg, 0.426 mmol) dissolved in DMF (1 mL) was added. After reacting at room temperature for 6 h, the mixture was moved to a 40 °C oil bath. During the reaction, 1 eq of HATU and 3 eq of TEA were added twice. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 10: 1), and the reaction was stopped after 20 h. H2O was added to the reaction solution, and the organic phase was concentrated and purified and separated by preparing plates for multiple times (DCM: MeOH = 10: 1). Compound A7 (33 mg, white solid, purity 95.3%) was obtained. Yield: 22.7%; LCMS (ESI) m / z calcd. for C 26 H 25 N5O[M+H] + 324.21found 324.2; 1H NMR (500MHz, DMSO-d6) δ10.14 (s, 1H), 9.58 (dd, J = 2.1, 0.9 Hz, 1H), 8.71 (d, J = 2.1 Hz, 1H), 8.34 (d, J = 2.3 Hz, 1H), 8.18 (d, J = 2.0 Hz, 1H), 7.52 (d, J = 8.1 Hz, 1H), 7.25–7.17 (m, 2H), 7.17–7.11 (m, 1H), 6.85 (dd, J = 2.3, 1.0 Hz, 1H), 6.53–6.47 (m, 1H), 3.00–2.92 (m, 2H), 2.90 (s, 6H), 2.52 (s, 3H), 1.33–1.27 (m, 3H). It can be seen that the product structure is correct.

[0159] Example 8

[0160] In this example, the compound 4-ethyl-N-[3-(imidazol-1-yl)phenyl]-3-[(2-methylpyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]benzamide (A8) was synthesized.

[0161]

[0162] Synthesis of Compound A8

[0163] A3-2 (100 mg, 0.328 mmol) was dissolved in DMF (5 mL), TEA (100 mg, 0.983 mmol) and HATU (125 mg, 0.328 mmol) were added, and after stirring for 5 min, A8-SM (68 mg, 0.426 mmol) dissolved in DMF (1 mL) was added, and the mixture was moved to a 30°C oil bath. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 20:1), and the reaction was stopped after 16 h. Water was added to the reaction solution, extracted with EA (20 ml x 3), dried over anhydrous Na2SO4, and the organic phase was concentrated. After column chromatography separation three times, compound A8 (15 mg, light yellow solid, purity 94.22%) was obtained. Yield: 9.7%; LCMS (ESI) m / z calcd.for C 27 H 22 N6O[M+H] + 447.19; found 447.2; 1H NMR (500MHz, DMSO-d6) δ10.55(s,1H),9.58(d,J=2.1Hz,1H),8.72(d,J=2.1Hz,1H),8. 35(d,J=2.4Hz,1H),8.26(s,1H),8.21(d,J=2.0Hz,1H),8.08(q,J=2.3Hz,1H),7.86–7 .79(m,1H),7.72(s,1H),7.59–7.49(m,2H),7.40(dd,J=7.9,2.2Hz,1H),7.19(s,1H),6.85(d,J=2.3Hz,1H),3.01–2.93(m,2H),2.48(s,3H),1.84(s,1H),1.34–1.28(m,3H). It can be seen that the product structure is correct.

[0164] Example 9

[0165] In this example, the compound 4-methoxy-3-{[2-(methylamino)pyrimidin-5-yl]ethynyl}-N-[3-(trifluoromethyl)phenyl]benzamide (Compound A9) was synthesized.

[0166]

[0167] Synthesis of Compound A9-1

[0168] Compound A9-SM1 (500 mg, 1.80 mmol), 3-trifluoromethylaniline (292 mg, 1.80 mmol), HATU (1026 mg, 2.70 mmol), DIEA (465 mg, 3.60 mmol) and DMF (5 mL) were added to a 50 mL single-mouth bottle in sequence. The reaction was carried out at 25 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into water (10 mL) and extracted three times with EA (15 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was separated by a preparative plate (PE / EA=2 / 1) to obtain compound A9-1 (100 mg, yellow solid, purity 94.898%). Yield: 14.07%; LCMS (ESI) m / zcalcd.for C 15 H 11 F3INO2[M+H] + 421.16 found 419.1. It can be seen that the product structure is correct.

[0169] Synthesis of Compound A9

[0170] Compound A9-1 (70 mg, 0.187 mmol), A9-SM2 (25 mg, 0.187 mmol), CuI (7.1 mg, 0.0373 mmol), Pd(PPh3)2Cl2 (26 mg, 0.0373 mmol), DIEA (48 mg, 0.373 mmol) and DMF (2 mL) were added to a 20 mL single-mouth bottle in sequence. The mixture was reacted at 40°C for 16 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (5 mL) to precipitate a solid, which was separated by high pressure preparation (column type: XBridge-1, 5 μm, 19-150 mm; detector: 254 nm; mobile phase: acetonitrile / water (0.1% formic acid solution); elution gradient: 40-75 / 8 min) to obtain compound A9 (16.7 mg, white solid, purity 99.3%). Yield: 20.9%; LCMS (ESI) m / z calcd.for C 22 H 17 F3N4O2[M+H] + 426.40; found 427.0. It can be seen that the product structure is correct.

[0171] Example 10

[0172] In this example, the compound N-{3-[(dimethylamino)methyl]-5-(trifluoromethyl)phenyl}-4-methoxy-3-{[2-(methylamino)pyrimidin-5-yl]ethynyl}benzamide (Compound A10) was synthesized.

[0173]

[0174] Synthesis of Compound A10-1

[0175] Compound A10-SM (130 mg, 0.593 mmol), A9-SM1 (165 mg, 0.593 mmol), HATU (338 mg, 0.889 mmol), DIEA (153 mg, 1.186 mmol) and DMF (5 mL) were added to a 50 mL single-mouth bottle in sequence. The reaction was carried out at 25 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (25 mL) and extracted three times with EA (40 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was separated by a preparative plate (PE / EA=2 / 1) to obtain compound A10-1 (50 mg, yellow solid, purity 95.038%). Yield: 16.73%; LCMS (ESI) m / z calcd.for C 18 H 18 F3IN2O2[M+H] +478.25 found 479.1. It can be seen that the product structure is correct.

[0176] Synthesis of Compound A10

[0177] Compound A10-1 (40 mg, 0.0835 mmol), A9-SM2 (22 mg, 0.167 mmol), CuI (3.2 mg, 0.0167 mmol), Pd(PPh3)2Cl2 (12 mg, 0.0167 mmol), DIEA (22 mg, 0.167 mmol) and DMF (1 mL) were added to a 20 mL single-mouth bottle in sequence. The mixture was reacted at 40°C for 16 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (5 mL) to precipitate a solid, which was separated by high pressure preparation (column type: XBridge-1, 5 microns, 19-150 mm; detector: 254 nm; mobile phase: acetonitrile / water (0.1% formic acid solution); elution gradient: 30-65 / 8 minutes), and filtered to obtain compound A10 (11.39 mg, yellow solid, purity 97.126%). Yield: 27.31%; LCMS (ESI) m / z calcd. for C 25 H 24 F3N5O2[M+H] + 483.50; found 484.2; 1 H NMR (400MHz, DMSO-d6): δ=10.45(s,1H),8.47(d,J=25.9Hz,2H),8.17(d,J=2.3Hz,2H),8.03(dd,J=8.7,2.4Hz,2H),7.63(q,J=4.7Hz,1H),7.34(s,1H),7.25(d,J=8.9Hz,1H),3.94(s,3H),3.48(s,2H),2.84(d,J=4.8Hz,3H),2.19(s,6H). It can be seen that the product structure is correct.

[0178] Embodiment 11

[0179] In this example, the compound N-[3-(dimethylamino)-5-(trifluoromethyl)phenyl]-4-methoxy-3-{[2-(methylamino)pyrimidin-5-yl]ethynyl}benzamide (Compound A11) was synthesized.

[0180]

[0181] Synthesis of Compound A11-1

[0182] Compound A11-SM (200 mg, 0.975 mmol), A9-SM1 (325 mg, 1.170 mmol), HATU (741 mg, 1.950 mmol), DIEA (504 mg, 3.899 mmol) and DMF (5 mL) were added to a 50 mL single-mouth bottle in sequence. The reaction was carried out at 25 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (25 mL) and extracted three times with EA (40 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was separated by a preparative plate (PE / EA=2 / 1) to obtain compound A11-1 (200 mg, yellow solid, purity 96.941%). Yield: 42.75%; LCMS (ESI) m / z calcd.for C17H 16 F3IN2O2[M+H] + 465.23found 465.1; 1 H NMR (400MHz,CDCl3):δ=8.21(s,1H),7.81(d,J=8.6Hz,1H),7.33(s,1H),7.19(s,1H),6.99(s,1H),6.81(d,J=8.6Hz,1H),6.62(s,1H),3.88(s,3H),2.95(s,6H). It can be seen that the product structure is correct.

[0183] Synthesis of Compound A11

[0184] Compound A11-1 (200 mg, 0.430 mmol), A9-SM2 (69 mg, 0.516 mmol), CuI (16 mg, 0.086 mmol), Pd(PPh3)2Cl2 (60 mg, 0.086 mol), DIEA (222 mg, 1.72 mmol) and DMF (2 mL) were added in sequence into a 50 mL single-necked bottle and reacted at 40 °C for 16 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (5 mL) to precipitate a solid. The solid was slurried with EA and DCM to obtain a filter cake. The filter cake was dissolved with DMSO and then dropped into water to precipitate a solid. The solid was separated by high pressure preparation (column type: XBridge-1, 5 μm, 19-150 mm; detector: 254 nm; mobile phase: acetonitrile / water (0.1% formic acid solution); elution gradient: 45-75 / 8 minutes) to obtain compound A10 (19.03 mg, yellow solid, purity 97.158%). Yield: 9.14%; LCMS (ESI) m / z calcd. for C 24 H 22 F3N5O2[M+H] +470.47;found470.3; 1 H NMR (400MHz, DMSO-d6): δ=10.23(s,1H),8.47(d,J=25.5Hz,2H),8.14(d,J=2.3Hz,1H),8.01(dd,J=8.8,2.3Hz,1H),7.67-7.59(m,1H),7.56(d,J=13.8Hz,1H),7.42(s,1H),7.24(d,J=8.9Hz,1H),6.67(s,1H),3.94(s,3H),2.97(s,6H),2.84(d,J=4.8Hz,3H). It can be seen that the product structure is correct.

[0185] Example 12

[0186] In this example, the compound 4-methoxy-N-[3-methoxy-5-(trifluoromethyl)phenyl]-3-{[2-(methylamino)pyrimidin-5-yl]ethynyl}benzamide (Compound A12) was synthesized.

[0187]

[0188] Synthesis of Compound A12-1

[0189] Compound A12-SM1 (300 mg, 1.56 mmol), A9-SM1 (521 mg, 1.87 mmol), HATU (1187 mg, 3.12 mmol), DIEA (807 mg, 6.24 mmol) and DMF (5 mL) were added to a 250 mL single-mouth bottle in sequence. The mixture was reacted at 25 °C for 0.5 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated aqueous sodium bicarbonate solution (25 mL) and extracted three times with EA (45 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was separated by a preparative plate (PE / EA=2 / 1) to obtain compound A12-1 (200 mg, yellow solid, purity 96.3%). Yield: 27.3%; LCMS (ESI) m / z calcd.for C 16 H 13 F3INO3[M+H] + 451.99found452.0. It can be seen that the product structure is correct.

[0190] Synthesis of Compound A12

[0191] Compound A12-1 (200 mg, 0.494 mmol), A9-SM2 (79 mg, 0.592 mmol), CuI (18.8 mg, 0.0987 mmol), Pd(PPh3)2Cl2 (69 mg, 0.0982 mol), DIEA (255 mg, 1.974 mmol) and DMF (5 mL) were added to a 50 mL single-necked bottle in sequence, and the mixture was reacted at 40° C. for 16 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (5 mL) to precipitate a solid. The solid was slurried with EA and DCM to obtain a filter cake. The filter cake was dissolved with DMSO and then dropped into water to precipitate a solid. The solid was dissolved with DMSO and separated by high pressure preparation (column type: XBridge-1, 5 μm, 19-150 mm; detector: 254 nm; mobile phase: acetonitrile / water (0.1% formic acid solution); elution gradient: 40-75 / 8 minutes) to obtain compound A12 (22.5 mg, yellow solid, purity 99.2%). Yield: 9.9%; LCMS (ESI) m / z calcd. for C 23 H 19 F3N4O3[M+H] + 457.14; found 457.2; 1 H NMR (400MHz, DMSO_d6): δ=10.42(s,1H),8.55-8.39(m,2H),8.15(d,J=2.3Hz,1H),8.01(dd,J=8.8,2.3Hz,1H),7.84(s,1H),7.74(s,1H),7.69-7.59(m,1H),7.26(d,J=8.9Hz,1H),6.97(s,1H),3.94(s,3H),3.84(s,3H),2.84(d,J=4.8Hz,3H). It can be seen that the product structure is correct.

[0192] Example 13

[0193] In this example, the compound 3-methoxy-5-{[(4-methoxy-3-{[2-(methylamino)pyrimidin-5-yl]ethynyl}phenyl)carbonyl]amino}benzene-1-carboxamide (Compound A13) was synthesized.

[0194]

[0195] Synthesis of Compound A13-1

[0196] A9-SM1 (850 mg, 3.06 mmol) was dissolved in DMF (10 mL), DIEA (1.2 g, 9.18 mmol) and HATU (1.1 g, 3.06 mmol) were added, and after stirring for 5 min, A13-SM (760 mg, 4.59 mmol) dissolved in DMF (5 mL) was added, and the mixture was moved to a 30°C oil bath. The reaction progress was monitored by TLC (developing solvent: PE:EA=8:1), and the reaction was stopped after 16 h. Water was added to the reaction solution, extracted with EA (20 mlx3), dried over anhydrous Na2SO4, and the organic phase was concentrated and column chromatography was performed to obtain compound A13-1 (600 mg, light yellow solid, 89.3%). Yield: 41.1%; LCMS (ESI) m / z calcd.for C 16 H 15 IN2O4[M+H] + 427.0 found 427.1. It can be seen that the product structure is correct.

[0197] Synthesis of Compound A13

[0198] Compound A13-1 (200 mg, 0.469 mmol), A9-SM2 (85 mg, 0.611 mmol), CuI (18 mg, 0.094 mmol), Pd(PPh3)2Cl2 (35 mg, 0.047 mmol), DIEA (180 mg, 1.407 mmol) and DMF (5 mL) were added to a 25 mL single-mouth bottle. The mixture was reacted at 80 °C for 5 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (1 mL) to precipitate a solid, which was then purified by a preparation plate (DCM: MeOH = 30: 1) to obtain compound A13 (51 mg, yellow solid, purity 95.1%). Yield: 23.9%; LCMS (ESI) m / z calcd. for C 23 H 21 N5O4[M+H] + 432.1; found 432.2. It can be seen that the product structure is correct.

[0199] Embodiment 14

[0200] In this example, compound 5-{[(4-methoxy-3-{[2-(methylamino)pyrimidin-5-yl]ethynyl}phenyl)carbonyl]amino}-3-(trifluoromethyl)benzene-1-carboxamide (Compound A14) was synthesized.

[0201]

[0202] Synthesis of Compound A14-1

[0203] A14-SM1 (500 mg, 2.6 mmol), A14-SM2 (500 mg, 2.6 mmol) and DMF (15 mL) were added to a 100 mL single-mouth bottle, and the solution turned dark orange-red. Pd(PPh3)2Cl2 (180 mg, 0.26 mmol), CuI (180 mg, 0.26 mmol), and DIEA (5 g, 39.4 mmol) were then added, and the color of the solution continued to deepen. The argon gas was replaced 3 times. Move it to an oil bath, heat it to 80 ° C and react for 12 hours, monitor the reaction progress by TLC (developing agent: PE: EA = 10: 1), and stop the reaction after the reaction is completed. Add about 30 mL of water and EA (30 mL × 3) for extraction, combine the organic phases, add anhydrous sodium sulfate to dry, and spin dry to obtain a crude product of 2 g of oily yellow dark red liquid. Column chromatography, eluent ratio PE:EA = 150:1 to 1:1, to obtain yellow solid compound A14-1 (300 mg, brown solid, 80%). Yield: 38.4%; LCMS (ESI) m / z calcd.for C 16 H 15 N3O3[M+H]+298.11; found 298.1. It can be seen that the product structure is correct.

[0204] Synthesis of Compound A14-2

[0205] In a 100mL single-mouth bottle, A14-1 (300mg, 1.01mmol) was dissolved in MeOH (10mL) and THF (10mL). The solution was light yellow and turbid. NaOH (3M) in H2O (10mL) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for reaction. TLC was used to monitor the progress of the reaction (developing solvent: PE:EA=4:1). After 16h, the reaction was completed and the reaction was stopped. The reaction solution was concentrated to remove most of the methanol and water. 1M HCl was used to adjust the pH to 4. A large amount of white solid precipitated, which was filtered and dried. Compound A14-2 (268mg, white solid) was obtained. Yield: 93.7%; the compound was used directly in the next step.

[0206] Synthesis of compound A14

[0207] A14-2 (250 mg, 0.88 mmol) was dissolved in DMF (5 mL), DIEA (350 mg, 2.64 mmol) and HATU (505 mg, 1.32 mmol) were added, and after stirring for 5 min, A13-SM (271 mg, 1.32 mmol) dissolved in 5 mL DMF was added, and the mixture was moved to a 30°C oil bath. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 20:1), and the reaction was stopped after 16 h. Water was added to the reaction solution, extracted with EA (20 ml x 3), dried over anhydrous Na2SO4, and the organic phase was concentrated. After column chromatography separation three times, compound A14 (32 mg, light yellow solid) was obtained. Yield: 7.8%. LCMS (ESI) m / z calcd.for C 23 H 18 F3N5O3[M+H] + 470.1; found 470.2. 1 H NMR (500MHz, DMSO-d6) δ10.60 (s, 1H), 8.57–8.49 (m, 2H), 8.48–8.40 (m, 2H), 8.24–8.18 (m, 2H), 8.05 (dd, J=8.8, 2.4 Hz, 1H), 7.96 (s, 1H), 7.65–7.58 (m, 2H), 7.27 (d, J=8.8 Hz, 1H), 3.95 (s, 3H), 2.85 (d, J=4.8 Hz, 3H). It can be seen that the product structure is correct.

[0208] Embodiment 15

[0209] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-[(2-{[1-(3-{[2-(2,6-dioxypiperidin-3-yl)-1-oxypiperidin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypiperidin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxybenzamide (Compound A15) was synthesized.

[0210]

[0211] Synthesis of Compound A15-1

[0212] A15-SM1 (5 g, 10.70 mmol) and DMF (80 mL) were added to a 100 mL single-mouth bottle, and trimethylsilyl acetylene (3.35 g, 34.20 mmol) was added dropwise, followed by Pd(PPh3)4 (1.19 g, 1.02 mmol), CuI (0.78 g, 4.11 mmol), and DIEA (12 g, 93 mmol). The argon gas was replaced three times, and the temperature was raised to 100 °C for reaction. The reaction progress was monitored by TLC (developing solvent: PE:EA=10:1). The reaction was stopped after 24 h, the solvent was concentrated, the residue was dissolved in ethyl acetate, 30 ml of saturated NaCl aqueous solution was added, and extracted with EA (30 ml×3), and the upper organic phase was added with anhydrous Na2SO4 for drying. After suction filtration, the crude product A15-1 (6.7 g, dark solid) was obtained by rotary evaporation.

[0213] Synthesis of Compound A15-2

[0214] A15-1 (1.1 g) was dissolved in THF (30 mL), and TBAF (1 M) in THF (30 mL) was added dropwise at 0°C. The reaction was allowed to react for 30 mins at room temperature. The reaction progress was monitored by TLC (developing solvent: PE: EA = 10:1). The reaction was stopped after 30 min. The mixture was quenched with water, extracted three times with ethyl acetate, and the upper organic phase was added with anhydrous Na2SO4 and dried. The crude product was obtained by filtration and rotary evaporation. Compound A15-2 (2.3 g, yellow solid) was obtained by column chromatography (PE: EA = 150:1 to 30:1). Yield: 70.7%.

[0215] Synthesis of Compound A15-3

[0216] A15-2 (1 g, 5.3 mmol), 5-chloro-2-iodopyrimidine (1.29 g, 5.3 mmol) and DMF (20 mL) were added to a 100 mL single-mouth bottle. The solution turned dark orange-red. Pd(PPh3)2Cl2 (0.19 g, 0.26 mmol) CuI (0.051 g, 0.26 mmol) and DIEA (2.7 g, 21 mmol) were then added. The color of the solution continued to deepen and the argon gas was replaced three times. After heating to 60 °C for 3 h, the reaction progress was monitored by TLC (developing solvent: PE:EA=5:1). The reaction was completed and the reaction was stopped. About 20 mL of water and EA (30 mLx3) were added for extraction. The organic phases were combined, dried with anhydrous sodium sulfate, and the crude product was spin-dried to obtain an oily yellow dark red liquid. After column chromatography, the eluent ratio was PE:EA = 100:1-10:1 to obtain compound A15-3 (1.2 g, white solid). Yield: 75.4%.

[0217] Synthesis of Compound A15-4

[0218] In a 50mL pressure tube, A15-3 (1.2g, 3.96mmol) was dissolved in isopropanol (30mL), 4-aminohexahydropyridine-1-carboxylic acid-2-methylpropane-2-yl ester (1.1g, 4.75mmol) was added, and DIEA (1.2g, 7.93mmol) was added. The solution was light yellow with a small amount of raw materials insoluble. The temperature was raised to 85°C, and after the reaction for 18h, the reaction progress was monitored by TLC (developing agent: PE:EA=2:1). The reaction was stopped when the reaction was completed. The solvent was concentrated, 20mL of water and DCM (30mLx3) were added for extraction, the organic phases were combined, anhydrous sodium sulfate was added for drying, and the crude product was spin-dried to obtain about 1.3g. After column chromatography (the eluent ratio was PE:EA=100:1~2:1), compound A15-4 (1.3g, beige solid) was obtained. Yield: 70.2%. LCMS (ESI) m / z calcd.for C 25 H 30 N4O5[M+H] + 467.22; found 467.2. It can be seen that the product structure is correct.

[0219] Synthesis of Compound A15-5

[0220] Add A15-4 (500 mg) to a 100 mL single-mouth bottle, dissolve in methanol (15 mL) and THF (15 mL), stir for 10 min, and the solution becomes transparent and clear. Slowly add NaOH (3 M, 9 mL) dropwise under ice bath conditions. Remove the ice bath and stir at room temperature for 10 min. React at room temperature for 2 h, monitor the reaction progress by TLC (developing solvent: PE: EA = 2:1), and stop the reaction when the reaction is complete. Concentrate the solvent, add about 20 mL of 1 M HCl aqueous solution under ice bath conditions to adjust the pH to 4-

[0221] 5. A large amount of solid precipitated, which was filtered and the filter cake was dried to obtain compound A15-5 (1 g, white solid).

[0222] Synthesis of Compound A15-6

[0223] A15-5 (1 g, 2.21 mmol) was added to a 25 mL single-mouth bottle, and DMF (15 mL) was added. The solution was white and turbid. TEA (856 mg, 6.63 mmol) and HATU (924 mg, 2.43 mmol) were then added. The solution was still turbid. A10-SM (620 mg, 2.43 mmol) was added, and the solution gradually turned orange and clear. The temperature was raised to 30 ° C, and the reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 10: 1). The reaction was stopped after 18 h. 30 mL of water and DCM (20 mL x 4) were added for extraction to obtain an organic phase, which was dried over anhydrous sodium sulfate and concentrated. Compound A15-6 (700 mg, white solid) was obtained by column chromatography purification (DCM: MeOH = 100:: 1 to 10: 1). Yield: 48.6%. LCMS (ESI) m / z calcd. for C 34 H 39 F3N6O4[M+H] + 653.30; found 653.3. It can be seen that the product structure is correct.

[0224] Synthesis of Compound A15-7

[0225] A15-6 (400 mg, 0.63 mmol) was added to a 50 mL single-mouth bottle and dichloromethane (5 mL). The solution was partially undissolved. HCl in 1,4-dioxane (4 M, 1.5 mL) was added dropwise. During the addition, the solution changed from turbid to clear, and then from clear to turbid. TLC was used to monitor the reaction progress (developing solvent: DCM: MeOH = 10:1). The reaction was stopped after 1 h. A large amount of solid precipitated in the reaction system. It was filtered (the filter cake was rinsed with DCM). The filter cake was dried to obtain compound A15-7 (320 mg, dark brown solid). Yield: 86.6%.

[0226] Synthesis of Compound A15

[0227] A15-7 (320 mg, 0.543 mmol) and A15-SM2 (236 mg, 0.797 mmol) were added to a 50 mL single-mouth bottle, and acetonitrile (10 mL) was added to partially dissolve. After adding potassium carbonate (225 mg, 1.630 mmol), the solution was still in a suspension state. The temperature was raised to 60 °C for reaction. TLC monitored the reaction progress (developing solvent: DCM: MeOH = 10:1), and the reaction was stopped after 16 hours. Concentrated, column chromatography, the eluent ratio was DCM: MeOH = 150:1 ~ 10:1, to obtain compound A15 (130 mg, off-white solid, 93.5%). Yield: 25.8%. LCMS (ESI) m / z calcd.for C 45 H 46F3N9O6[M+H] + 866.35; found 866.4; 1 H NMR(600MHz,DMSO-d6)δ10.98(s,1H),10.58(s,1H),10.49(s,1H),8.47(s,2H),8.21–8.16(m,2H),8.05(d,J=6.8Hz,2H),8 .01(s,1H),7.72–7.64(m,2H),7.62(d,J=8.7Hz,1H),7.35(s,1H),7.25(d,J=8.7Hz,1H),5.09(dd,J=13.3,5.0Hz,1H),4.44 (d, J = 17.0 Hz, 1H), 4.30 (d, J = 17.1 Hz, 1H), 3.94 (s, 3H), 3.78 (s, 1H), 3.50 (d, J = 10.6 Hz, 2H), 2.99–2.88 (m, 3H), 2.70 (s, 2H), 2.64–2.55 (m, 3H), 2.43–2.36 (m, 1H), 2.20 (s, 8H), 2.00 (d, J = 13.2 Hz, 1H), 1.87 (d, J = 10.2 Hz, 2H), 1.56 (d, J = 12.2 Hz, 2H). It can be seen that the product structure is correct.

[0228] Example 16

[0229] In this example, compound 3-[({3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-ethylphenyl}carbonyl)amino]-5-(trifluoromethyl)benzene-1-carboxamide (Compound A16) was synthesized.

[0230]

[0231] Synthesis of Compound A16-1

[0232] A3-SM1 (1 g, 5.3 mmol), 5-chloro-2-iodopyrimidine (1.29 g, 5.3 mmol) were added to a 100 mL single-mouth bottle. DMF (20 mL) was added, and the solution turned dark orange-red. Pd (PPh3) 2Cl2 (0.19 g, 0.26 mmol) CuI (0.051 g, 0.26 mmol) and DIEA (2.7 g, 26 mmol) were then added. The color of the solution continued to deepen, and the argon gas was replaced 3 times. After heating to 80 ° C for 12 hours, the reaction progress was monitored by TLC (developing solvent: PE: EA = 10: 1). The reaction was completed and the reaction was stopped. About 20 mL of water and EA (30 mL x 3) were added for extraction, the organic phases were combined, anhydrous sodium sulfate was added for drying, and the crude product was spin-dried to obtain an oily yellow dark red liquid. After column chromatography, the eluent ratio was PE:EA = 150:1-50:1 to obtain compound A16-1 (900 mg, yellow solid). Yield: 56.4%. 1 H NMR (500MHz, DMSO-d6) δ9.06 (s, 2H), 8.12 (d, J = 1.9 Hz, 1H), 7.98 (dd, J = 8.1, 1.9 Hz, 1H), 7.55 (d, J = 8.1 Hz, 1H), 3.89 (s, 3H), 2.94 (q, J = 7.5 Hz, 2H), 1.28 (t, J = 7.5 Hz, 3H). It can be seen that the product structure is correct.

[0233] Synthesis of Compound A16-2

[0234] A16-1 (450 mg, 1.5 mmol) was dissolved in isopropanol (10 mL) in a 50 mL pressure tube, 4-aminopiperidin-1-carboxylic acid-2-methylpropane-2-yl ester (380 mg, 1.9 mmol) was added, and DIEA (387 mg, 3 mmol) was added. The solution was light yellow with a small amount of insoluble raw materials. The temperature was raised to 85 ° C. After the reaction was carried out for 18 hours, the reaction progress was monitored by TLC (developing solvent: PE: EA = 2: 1). The reaction was stopped when the reaction was completed. The solvent was concentrated, 20 mL of water and DCM (20 mL x 3) were added for extraction, the organic phases were combined, anhydrous sodium sulfate was added for drying, and the crude product was spin-dried to obtain about 1.3 g. After column chromatography (the eluent ratio was PE: EA = 100: 1 to 2: 1), compound A16-2 (610 mg, light yellow oil) was obtained. Yield: 87.7%. 1H NMR (500MHz, DMSO-d6) δ8.51 (s, 2H), 7.88 (dd, J = 8.0, 1.9 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.48 (d, J = 8.1 Hz, 1H), 4.09 (q, J = 5.3 Hz, 1H), 3.85 (s, 3H), 3.32 (s, 1H), 3.17 (d, J = 5.2 Hz, 3H), 2.87 (q, J = 7.5 Hz, 3H), 1.83 (dd, J = 13.0, 3.7 Hz, 2H), 1.40 (s, 9H), 1.37 (dd, J = 11.8, 4.4 Hz, 2H), 1.24 (t, J = 7.5 Hz, 3H). It can be seen that the product structure is correct.

[0235] Synthesis of Compound A16-3

[0236] Add A16-2 (500 mg, 1.08 mmol) to a 100 mL single-mouth bottle and dissolve in methanol (27 mL). The raw material has poor solubility. Stir for 10 min and the solution becomes transparent and clear. Slowly add NaOH (2M, 9 mL) dropwise under ice bath conditions. Remove the ice bath and stir at room temperature for 10 min. Heat to 40 ° C for 2 h. Monitor the reaction progress by TLC (developing solvent: PE: EA = 2: 1). The reaction is complete and the reaction is stopped. Concentrate the solvent and add about 16 mL of 1M HCl aqueous solution under ice bath conditions to adjust the pH to 4-5. A large amount of solid precipitates. Filter and dry the filter cake to obtain compound A16-3 (450 mg, white solid) with a yield of 92.8%.

[0237] Synthesis of Compound A16-4

[0238] A16-3 (300 mg, 0.67 mmol) was added to a 25 mL single-mouth bottle, and DMF (5 mL) was added. The solution was white and turbid. TEA (200 mg, 0.67 mmol) and HATU (250 mg, 0.67 mmol) were added. The solution was still turbid. 3-amino-5-trifluoromethylbenzamide (160 mg, 0.67 mmol) was added, and the solution gradually turned orange and clear. The temperature was raised to 30 ° C, and the reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 10: 1). The reaction was stopped after 18 hours. 10 mL of water and DCM (10 mL x 4) were added for extraction to obtain an organic phase, which was dried over anhydrous sodium sulfate and concentrated. The compound A16-4 (150 mg, white solid) was purified by preparative plate (DCM: MeOH = 10: 1) with a yield of 35.4%.

[0239] Synthesis of Compound A16-5

[0240] A16-4 (150 mg, 0.26 mmol) was added to a 50 mL single-mouth bottle. Dichloromethane (5 mL) was added. The solution was partially undissolved. HCl in 1,4-dioxane (4 M, 1 mL) was added dropwise. During the addition, the solution changed from turbid to clear, and then from clear to turbid. TLC was used to monitor the reaction progress (developing solvent: DCM: MeOH = 10:1). The reaction was stopped after 1 h. A large amount of solid precipitated in the reaction system. It was filtered (the filter cake was rinsed with DCM). The filter cake was dried to obtain compound A16-5 (100 mg, dark yellow solid). The yield was 74.1%.

[0241] Synthesis of Compound A16

[0242] A16-5 (100 mg, 0.175 mmol) and A15-SM2 (76 mg, 0.192 mmol) were added to a 50 mL single-mouth bottle, and acetonitrile (5 mL) was added to partially dissolve. After adding potassium iodide (32 mg, 0.192 mmol) and potassium carbonate (72 mg, 0.524 mmol), the solution was still in a suspension state. The temperature was raised to 80 °C for reaction. TLC monitored the reaction progress (developing solvent: DCM: MeOH = 10:1), the reaction was stopped after 6 hours, concentrated, and column chromatography was performed with an eluent ratio of DCM: MeOH = 150:1 to 10:1 to obtain compound A16 (20 mg, yellow solid, 94%). Yield: 12.8%. LCMS (ESI) m / z calcd.for C 44 H 42 F3N9O6[M+H] + 850.32;found850.6; 1H NMR(600MHz,DMSO-d6)δ10.97(s,1H),10.72(s,1H),10.56(s,1H),8.57(s,1H),8.51(s,2H),8.43(s,1H),8.23(s,1H),8.15(s,1H),7.99(s,1 H),7.97–7.92(m,2H),7.75(d,J=6.1Hz,1H),7.66(d,J=8.3Hz,1H),7.63–7.59(m,2H),7.51(d,J=8.1Hz,1H),5.08(dd,J=13.3,5.0Hz,1H),4.4 3 (d, J = 17.2 Hz, 1H), 4.29 (d, J = 17.2 Hz, 1H), 4.10 (d, J = 5.1 Hz, 1H), 3.80 (s, 1H), 3.17 (d, J = 4.6 Hz, 2H), 3.03–2.85 (m, 5H), 2.74 (s, 1H), 2.60 (d, J = 16.4 Hz, 3H), 2.42–2.33 (m, 1H), 2.18 (t, J = 5.9 Hz, 1H), 2.02–1.97 (m, 1H), 1.88 (d, J = 10.9 Hz, 2H), 1.58 (d, J = 8.0 Hz, 2H), 1.34–1.27 (m, 3H). It can be seen that the product structure is correct.

[0243] Embodiment 17

[0244] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-[(2-{[4-(4-{[2-(2,6-dioxypyridin-3-yl)-1-oxyylidene-2,3-dihydro-1H-isoindol-5-yl]methyl}piperazin-1-yl)-1-oxyylidenebutyl]amino}pyrimidin-5-yl)ethynyl]-4-methoxybenzamide (Compound A17) was synthesized.

[0245]

[0246] Synthesis of Compound A17-1

[0247] A17-SM1 (3 g, 9.28 mmol) was dissolved in DMF (60 mL) at 0 °C, and A17-SM2 (3.6 g, 11.14 mmol), palladium acetate (110 mg, 0.928 mmol), Xphos (450 mg, 1.86 mmol) and sodium carbonate (4.5 g, 41.8 mmol) were added. The temperature was raised to 90 °C for reaction. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 10:1). The reaction was stopped after 16 h, ethyl acetate (50 mL) was added and stirred, and the filtrate was filtered to obtain the filtrate, extracted with water, and the upper organic phase was added with anhydrous Na2SO4 to dry. The crude product was then filtered and evaporated. Compound A17-1 (2.3 g, yellow solid) was obtained by column chromatography (PE: EA = 150:1-30:1), with a yield of 55.9%.

[0248] Synthesis of Compound A17-2

[0249] Compound A17-1 (2.3 g, 5.2 mmol) and DCM (30 mL) were added to a 100 mL single-mouth bottle, followed by 4 N HCl in dioxane (8 mL). The mixture was reacted at 25 °C for 2 hours under nitrogen protection. After the reaction was completed, the mixture was concentrated to dryness to obtain a crude compound A17-2 (1.7 g, yellow solid). Yield: 91.7%. LCMS (ESI) m / z calcd. for C18H22N4O3 [M+H] + 343.2; found 343.3.

[0250] Synthesis of Compound A17-3

[0251] A17-2 (300 mg, 0.876 mmol) and 4-chlorobutylamide (120 mg, 1.05 mmol) were added to a 50 mL single-mouth bottle, and acetonitrile (15 mL) was added to partially dissolve. After adding potassium carbonate (360 mg, 2.63 mmol), the solution was still in a suspension state. The temperature was raised to 60 °C for reaction. TLC monitored the reaction progress (developing solvent: DCM: MeOH = 5: 1), and the reaction was stopped after 24 hours. The reaction solution was filtered and the filtrate was concentrated. After column chromatography, the eluent ratio was DCM: MeOH = 100: 1 to 10: 1 to obtain A17-3 (120 mg, off-white solid). Yield: 32.0%. LCMS (ESI) m / z calcd. For C22H29N5O4 [M + H] + 428.2; found 428.3. It can be seen that the product structure is correct.

[0252] Synthesis of Compound A17-4

[0253] Add A10-1 (1g, 5.3mmol), 2-chloro-5-ethynylpyrimidine (350mg, 5.3mmol) and DMF (20mL) to a 100mL single-mouth bottle. The solution turns dark orange-red. Then add Pd(PPh3)2Cl2 (0.19g, 0.26mmol) CuI (0.051g, 0.26mmol) and TEA (2.7g, 26mmol). The color of the solution continues to deepen and the argon gas is replaced 3 times. After heating to 60℃ for 3h, the reaction progress is monitored by TLC (developing solvent: PE:EA=5:1). The reaction is complete and the reaction is stopped. Add about 20mL of water and EA (30mLx3) for extraction, combine the organic phases, add anhydrous sodium sulfate to dry, and spin dry to obtain the crude product as an oily yellow dark red liquid. After column chromatography, the eluent ratio was PE:EA = 100:1-10:1 to obtain compound A17-4 (1.2 g, off-white solid) with a yield of 77.4%.

[0254] Synthesis of Compound A17

[0255] Compound A17-4 (200 mg, 0.410 mmol), A17-3 (210 mg, 0.491 mmol) and 10 mL 1,4-dioxane were added to a 5 mL single-mouth bottle and stirred at room temperature for 10 min. Cesium carbonate (300 mg, 0.820 mmol), Xantphos (50 mg, 0.082 mmol) and Pd2(dba)3 (74 mg, 0.041 mmol) were added. The mixture was reacted at 100 °C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was cooled to room temperature, 15 mL of purified water was added and stirred for 8 min, and ethyl acetate (15 mL × 2) was used for extraction. The ethyl acetate phases were combined, the organic phases were dried and concentrated, and purified by column chromatography (DCM: MeOH = 150: 1-151) to obtain compound A17 (53 mg, yellow solid, yield: 53%). LCMS (ESI) m / z calcd.for C 46 H 48 F3N9O6[M+H]+880.4; found880.5. It can be seen that the product structure is correct.

[0256] Embodiment 18

[0257] In this example, compound 3-[({3-[(2-{[4-(4-{[2-(2,6-dioxypyridin-3-yl)-1-oxyylidene-2,3-dihydro-1H-isoindol-5-yl]methyl}piperazin-1-yl)-1-oxyylidenebutyl]amino}pyrimidin-5-yl)ethynyl]-4-ethylphenyl}carbonyl)amino]-5-(trifluoromethyl)benzene-1-carboxamide (Compound A18) was synthesized.

[0258]

[0259] Synthesis of Compound A18-1

[0260] A2-1 (1 g, 3.62 mmol) was dissolved in DMF (10 mL), DIEA (1.4 g, 10.86 mmol) and HATU (1.4 g, 3.62 mmol) were added, and after stirring for 5 min, 3-amino-5-trifluoromethylbenzamide (740 mg, 3.62 mmol) was added, and the mixture was moved to a 30°C oil bath. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 20:1), and the reaction was stopped after 16 h. The reaction solution was added with water, extracted with EA (20 ml x 3), dried over anhydrous Na2SO4, and the organic phase was concentrated. After column chromatography separation, compound A18-1 (159 mg, light yellow solid, 90.3%) was obtained. Yield: 59.3%. LCMS (ESI) m / z calcd. for C 17 H 14 F3IN2O2[M+H] + 463.1; found 463.2. It can be seen that the product structure is correct.

[0261] Synthesis of compound A18-2

[0262] Add A18-1 (1g, 5.3mmol), 2-chloro-5-ethynylpyrimidine (350mg, 5.3mmol) and DMF (20mL) to a 100mL single-mouth bottle. The solution turns dark orange-red. Then add Pd(PPh3)2Cl2 (0.21g, 0.26mmol) CuI (0.053g, 0.26mmol) and DIEA (2.7g, 26mmol). The color of the solution continues to deepen and the argon gas is replaced 3 times. After heating to 60℃ for 3h, the reaction progress is monitored by TLC (developing solvent: PE:EA=5:1). The reaction is complete and the reaction is stopped. Add about 20mL of water and EA (30mLx3) for extraction, combine the organic phases, add anhydrous sodium sulfate to dry, and spin dry to obtain the crude product as an oily yellow dark red liquid. After column chromatography, the eluent ratio was PE:EA=100:1-15:1 to obtain compound A18-2 (1.1 g, off-white solid). Yield: 71.3%.

[0263] Synthesis of Compound A18

[0264] Compound A18-2 (200 mg, 0.411 mmol), A17-3 (210 mg, 0.493 mmol) and 10 mL 1,4-dioxane were added to a 25 mL single-mouth bottle, stirred at room temperature for 10 min, and the reaction solution was heterogeneous. Cesium carbonate (304 mg, 0.822 mmol), Xantphos (50 mg, 0.082 mmol), and Pd2(dba)3 (74 mg, 0.041 mmol) were added. The reaction was carried out at 100 ° C for 16 hours under nitrogen protection. The reaction solution was cooled to room temperature, 15 mL of purified water was added and stirred for 8 min, and ethyl acetate (15 mL × 2) was used for extraction. The ethyl acetate phases were combined, the organic phases were dried and concentrated, and purified by column chromatography (DCM: MeOH = 150: 1-151) to obtain compound A18 (43 mg, yellow solid, purity: 97.8%) with a yield of 12.1%. LCMS (ESI) m / z calcd.for C 45 H 44 F3N9O6[M+H]+864.3; found 864. It can be seen that the product structure is correct.

[0265] Embodiment 19

[0266] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-[(2-{[1-(3-{[2-(2,6-dioxypiperidin-3-yl)-1-oxypiperidin-2,3-dihydro-1H-isoindol-4-yl]amino}-3-oxypiperidin-4-yl)acetyl]-4-methoxybenzamide (Compound A19) was synthesized.

[0267]

[0268] Synthesis of Compound A19-1

[0269] A19-SM (1 g, 5.3 mmol) was dissolved in THF (15 mL), stirred in an ice bath at 0°C and 3-bromopropionyl chloride was slowly added dropwise. After 20 min, the ice bath was removed and moved to an oil bath with the temperature set at 50°C. The reaction progress was monitored by TLC (developing solvent: DCM:MeOH=40:1). The reaction was stopped after 5 h. About 5 mL of methanol was added, stirred for 30 min, filtered, and the filter cake was dried to obtain compound A19-1 (890 mg, yellow solid, purity: 91.8%) with a yield of 92.1%. 1H NMR (500MHz, DMSO-d6) δ11.03 (s, 1H), 9.98 (s, 1H), 7.89–7.78 (m, 1H), 7.58–7.47 (m, 2H), 5.15 (dd, J = 13.3, 5.1 Hz, 1H), 4.37 (q, J = 17.5 Hz, 2H), 3.75 (t, J = 6.4 Hz, 2H), 3.01 (t, J = 6.4 Hz, 2H), 2.96–2.88 (m, 1H), 2.65–2.58 (m, 1H), 2.33 (qd, J = 13.1, 4.4 Hz, 1H), 2.04 (dtd, J = 12.9, 5.4, 2.4 Hz, 1H). It can be seen that the product structure is correct.

[0270] Synthesis of compound A19

[0271] A19-1 (300 mg, 5.3 mmol) and A15-7 (360 mg, 5.3 mmol) were placed in a pressure tube, acetonitrile (5 mL) was added, and then KI (93 mg, 1.3 mmol) and K2CO3 (211 mg, 2.6 mmol) 11 mg were added. T = 80 ° C, TLC was used to monitor the reaction progress (developing solvent: DCM: MeOH = 30: 1), and the reaction was stopped after 18 hours. The solution was turbid, filtered, and the filtrate was concentrated and TLC was prepared (DCM: MeOH = 8: 1) to obtain an oily substance, which was slurried with isopropanol to obtain compound A19 (160 mg, yellow solid, purity: 88.7%) with a yield of 45.1%. LCMS (ESI) m / z calcd. for C 45 H 47 F3N9O6[M+H] + 866.3; found 866.1. 1HNMR(500MHz,DMSO-d6)δ11.02(s,1H),10.45(s,1H),10.03(s,1H),8.47(s,2H),8.20–8.16(m,2H),8.06–8.01(m,2H) ,7.84(dd,J=7.0,1.6Hz,1H),7.65(d,J=7.7Hz,1H),7.54–7.47(m,2H),7.34(s,1H),7.25(d,J=8.9Hz,1H),5.15(dd,J =13.3,5.1Hz,1H),4.44–4.32(m,2H),3.94(s,3H),3.78(d,J=15.5Hz,1H),3.48(s,2H),2.98–2.88(m,3H),2.71–2.53(m,5H),2.35(qd,J=13.2,4.4Hz,1H),2.19(s,6H),2.15–2.00(m,3H),1.89(d,J=15.2Hz,2H),1.54(d,J=12.0Hz,2H). It can be seen that the product structure is correct.

[0272] Embodiment 20

[0273] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]oxy}propyl)pyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxybenzamide (Compound A20) was synthesized.

[0274]

[0275] Synthesis of Compound A20-1

[0276] A20-SM (2 g, 7.68 mmol) and 3-bromo-1-propanol (2.67 g, 19.21 mmol) were added to a 50 mL single-mouth bottle, and DMF (15 mL) was added to partially dissolve. After adding potassium bicarbonate (2.3 g, 23.04 mmol), the solution was still in a suspension state. The temperature was raised to 60 °C for reaction. TLC monitored the reaction progress (developing solvent: DCM: MeOH = 10: 1), and the reaction was stopped after 24 hours. The reaction solution was filtered, and the filtrate was added with 5% acetic acid aqueous solution, extracted with DCM three times, and PE and EA were added to pulp until the product was a white solid. After rotary evaporation, acetonitrile was added to pulp, and the filter cake was filtered and dried to obtain compound A20-1 (750 mg, white solid). Yield: 30.7%. LCMS (ESI) m / z calcd. For C 16 H18 N2O5[M+H] + 319.1; found 319.3. It can be seen that the product structure is correct.

[0277] Synthesis of Compound A20-2

[0278] A20-1 (1.5 g, 4.71 mmol) was dissolved in 25 mL DMF under an ice-water bath, and the solution was yellow and clear. Dess-Martin periodinane (3 g, 7.07 mmol) was added to the reaction flask. The reaction was carried out at room temperature under argon protection. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 20: 1), and the reaction was stopped after 2 h. Under an ice-water bath, the reaction solution was poured into a beaker containing a saturated NaHCO3 solution, and an orange-yellow solid was precipitated. After stirring for 10 mins, it was extracted with EA, dried and concentrated to obtain compound A20-2 (1.1 g, yellow solid) with a yield of 73.3%.

[0279] Synthesis of compound A20

[0280] A20-2 (100 mg, 0.316 mmol) and A5-7 (170 mg, 0.316 mmol) were dissolved in 5 ml dry DMF, AcOH (38 mg, 0.632 mmol) and STAB (135 mg, 0.632 mmol) were added, ventilated, and reacted under argon protection for 24 h. In an ice-water bath, the reaction solution was poured into a beaker containing a saturated NaHCO3 solution, stirred for 10 mins, extracted with DCM, dried and concentrated to obtain a crude product. Column chromatography purification (DCM: MeOH = 150: 1 ~ 10: 1) gave compound A20 (51 mg, off-white solid, purity: 95.8%). Yield: 18.1%. LCMS (ESI) m / z calcd.for C 45 H 47 F3N8O6[M+H] + 853.4; found 853.5. It can be seen that the product structure is correct.

[0281] Embodiment 21

[0282] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-4-yl]oxy}propyl)pyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxybenzamide (Compound A21) was synthesized.

[0283]

[0284] Synthesis of Compound A21-1

[0285] A21-SM (2 g, 7.68 mmol) and 3-bromo-1-propanol (2.66 g, 19.2 mmol) were added to a 50 mL single-mouth bottle, and DMF (15 mL) was added. Potassium bicarbonate (2.3 g, 23.04 mmol) was added and the temperature was raised to 60°C for reaction. TLC was used to monitor the reaction progress (developing solvent: DCM: MeOH = 10:1). The reaction was stopped after 24 hours. The reaction solution was filtered, and the filtrate was added with 5% acetic acid aqueous solution, extracted with DCM three times, PE and EA were added to slurry until the product was a white solid, and acetonitrile was added to slurry after rotary evaporation, and the filter cake was filtered and dried to obtain compound A21-1 (920 mg, white solid). Yield: 37.7%. LCMS (ESI) m / z calcd.For C 16 H 18 N2O5[M+H] + 319.1; found 319.3. It can be seen that the product structure is correct.

[0286] Synthesis of Compound A21-2

[0287] A21-1 (1.5 g, 4.71 mmol) and 25 mL DMF were added to a 100 mL three-necked round-bottom flask under an ice-water bath, and the solution was yellow and clear. Dess-Martin periodinant (3 g, 7.07 mmol) was added to the reaction flask. The reaction was carried out at room temperature under argon protection. TLC monitored the progress of the reaction (developing solvent: DCM: MeOH = 20: 1), and the reaction was stopped after 2 h. Under an ice-water bath, the reaction solution was poured into a beaker containing a saturated NaHCO3 solution, and an orange-yellow solid was precipitated. After stirring for 10 mins, it was extracted with EA, dried and concentrated to obtain compound A21-2 (960 mg, yellow solid) with a yield of 63.9%.

[0288] Synthesis of compound A21

[0289] A21-2 (100 mg, 0.316 mmol) and A15-7 (170 mg, 0.316 mmol) were dissolved in 5 ml dry DMF, AcOH (38 mg, 0.632 mmol) and STAB (135 mg, 0.632 mmol) were added, ventilated, and reacted for 24 h under argon protection. In an ice-water bath, the reaction solution was poured into a beaker containing a saturated NaHCO3 solution, stirred for 10 mins, extracted with DCM, dried and concentrated to obtain a crude product. Column chromatography purification (DCM: MeOH = 150: 1 ~ 10: 1) gave compound A21 (41 mg, off-white solid, purity: 95.8%). Yield: 14.0%. LCMS (ESI) m / z calcd.for C45 H 47 F3N8O6[M+H] + 853.4; found 853.5. It can be seen that the product structure is correct.

[0290] Embodiment 22

[0291] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-[(5-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypyridin-4-yl]amino}pyrazin-2-yl)ethynyl]-4-methoxybenzamide (Compound A22) was synthesized.

[0292]

[0293] Synthesis of Compound A22-1

[0294] A14-SM1 (1g, 5.3mmol), A22-SM1 (350mg, 6.3mmol) were added to a 100mL single-mouth bottle, and DMF (20mL) was added. The solution turned dark orange-red, and then Pd(PPh3)2Cl2 (0.19g, 0.26mmol) CuI (0.051g, 0.26mmol), and DIEA (2.7g, 26mmol) were added. The color of the solution continued to deepen, and the argon gas was replaced 3 times. After heating to 100℃ for 16h, the reaction progress was monitored by TLC (developing agent: PE:EA=5:1). The reaction was completed and the reaction was stopped. About 20mL of water and EA (30mLx3) were added for extraction, the organic phases were combined, anhydrous sodium sulfate was added for drying, and the crude product was spin-dried to obtain an oily yellow dark red liquid. After column chromatography, the eluent ratio was PE:EA=

[0295] 100:1~10:1, to obtain compound A22-1 (1.2 g, off-white solid), yield: 72.8%.

[0296] Synthesis of Compound A22-2

[0297] A22-1 (1g, 3.53mmol) and N-Boc-piperidin-4-one (704mg, 3.53mmol) were dissolved in 5ml dry DMF, AcOH (200mg, 3.53mmol) and STAB (1.1g, 5.3mmol) were added, and the mixture was ventilated and reacted under argon protection for 24h. In an ice-water bath, the reaction solution was poured into a beaker containing a saturated NaHCO3 solution, stirred for 10mins, extracted with DCM, dried and concentrated to obtain a crude product. Column chromatography purification (PE:EA=4:1) obtained an off-white solid compound (640mg), and this intermediate was hydrolyzed with NaOH (2M) in 30mL methanol to obtain the carboxylic acid derivative A22-2 (purity: 91%). Yield: 35%. LCMS (ESI) m / zcalcd.for C 24 H 29 N4O5[M+H] + 453.2; found 453.3. It can be seen that the product structure is correct.

[0298] Synthesis of Compound A22-3

[0299] A22-2 (1 g, 2.21 mmol) was added to a 25 mL single-mouth bottle, and DMF (15 mL) was added. The solution was white and turbid. TEA (500 mg, 4.42 mmol) and HATU (680 mg, 2.21 mmol) were then added. The solution was still turbid. A10-SM (580 mg, 2.65 mmol) was added. The temperature was raised to 30 °C, and the reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 15:1). After 18 h, the reaction solution was poured into 30 mL of water, filtered to obtain a filter cake, dried, and 10 mL of methanol was added to slurry and filter to obtain a filter cake. Compound A22-3 (1.1 g, off-white solid, purity: 92.1%) was obtained by drying. Yield: 70.2%. LCMS (ESI) m / z calcd. For C 34 H 39 F3N6O4[M+H] + 653.3; found 653.4. It can be seen that the product structure is correct.

[0300] Synthesis of Compound A22-4

[0301] A22-3 (600 mg, 0.94 mmol) was added to a 50 mL single-mouth bottle and dichloromethane (20 mL). The solution was partially undissolved. HCl in 1,4-dioxane (4 M, 4 mL) was added dropwise. During the addition, the solution changed from turbid to clear, and then from clear to turbid. TLC was used to monitor the reaction progress (developing solvent: DCM: MeOH = 15:1). The reaction was stopped after 4 hours. A large amount of solid precipitated in the reaction system. It was filtered (the filter cake was rinsed with DCM), and the filter cake was dried to obtain compound A22-4 (550 mg, yellow solid). Yield: 94.3%. LCMS (ESI) m / z calcd. for C 29 H 31 F3N6O2[M+H] + 553.3; found 553.4. It can be seen that the product structure is correct.

[0302] Synthesis of compound A22

[0303] A22-4 (300 mg, 0.54 mmol) and A15-SM2 (220 mg, 0.54 mmol) were added to a 50 mL single-mouth bottle, and acetonitrile (15 mL) was added to partially dissolve. After adding potassium carbonate (250 mg, 1.62 mmol), the solution was still in a suspension state. The temperature was raised to 60 °C for reaction. TLC monitored the reaction progress (developing solvent: DCM: MeOH = 10: 1), and the reaction was stopped after 24 hours. The reaction solution was filtered and the filtrate was concentrated. After column chromatography, the eluent ratio was DCM: MeOH = 100: 1 to 10: 1 to obtain 180 mg of yellow solid. Compound A22 (50 mg, off-white solid, 95.7%) was obtained by preparation plate. Yield: 11.6%. LCMS (ESI) m / z calcd. For C 45 H 46 F3N9O6[M+H] + 866.4; found 866.5. 1H NMR(600MHz,DMSO-d6)δ10.8(s,1H),10.5(s,1H),10.3(s,1H),8.37(s,2H),8.19–8.13(m,2H),8.05(d,J=6.8Hz,2H), 8.0(s,1H),7.67–7.61(m,2H),7.6(d,J=8.3Hz,1H),7.25(s,1H),7.15(d,J=8.7Hz,1H),5.29(dd,J=13.3,5.0Hz,1H),4 .37(d,J=17.0Hz,1H),4.25(d,J=17.1Hz,1H),3.86(s,3H),3.72(s,1H),3.52(d,J=10Hz,2H),3.09–2.9(m,3H),2.72(s,2H),2.62–2.52(m,3H),2.40–2.31(m,1H),2.1(s,8H),1.9(d,J=12Hz,1H),1.77(d,J=9Hz,2H),1.36(d,J=12Hz,2H). It can be seen that the product structure is correct.

[0304] Embodiment 23

[0305] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-({2-[(2-{4-[4-(2,4-dioxyhexahydropyrimidin-1-yl)phenyl]hexahydropyridin-1-yl}ethyl)amino]pyrimidin-5-yl}ethynyl)-4-methoxybenzamide (Compound A23) was synthesized.

[0306]

[0307] Synthesis of Compound A23-1

[0308] A23-SM1 (10 g, 37.16 mmol), A23-SM2 (17 g, 55.74 mmol), 1,4-dioxane (100 mL) and water (20 mL) were added to a 500 mL single-mouth bottle, potassium carbonate (10.3 g, 74.32 mmol), Pd(dppf)Cl2 (1.4 g, 1.86 mmol), and the temperature was raised to 90°C for reaction after argon replacement. The reaction was stopped after 7 h, the solvent was added with water, extracted with ethyl acetate three times, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, the residue was slurried with PE:EA=2:1 (50 mL), filtered to obtain a filter cake, and dried to obtain compound A23-1 (7.7 g, off-white solid), with a yield of 49.7%.

[0309] Synthesis of Compound A23-2

[0310] A23-1 (7.7 g, 20.73 mmol) was added to a 500 mL single-mouth bottle, dissolved in EtOH (150 mL), and Pd / C (0.3 g) was added. The reaction solution was stirred overnight at room temperature in a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered through diatomaceous earth and the filtrate was dried by spin drying. The crude product was purified by column chromatography (DCM / MeOH=100 / 1-10 / 1) to obtain compound A23-2 (5.7 g, gray solid, purity: 90.3%). Yield: 66.4%. LCMS (ESI) m / z calcd.for C 20 H 27 N3O4[M+H] + 374.2; found 374.3. It can be seen that the product structure is correct.

[0311] Synthesis of Compound A23-3

[0312] Compound A23-2 (2.3 g, 5.19 mmol) and DCM (30 mL) were added to a 100 mL single-mouth bottle, followed by 4N HCl in dioxane (8 mL). The mixture was reacted at 25 °C for 2 hours under nitrogen protection. After the reaction was completed, the mixture was concentrated to dryness to obtain a crude compound A23-3 (1.7 g, yellow solid). Yield: 90.2%. LCMS (ESI) m / z calcd. for C 15 H 19 N3O2[M+H] + 274.2; found 274.2. It can be seen that the product structure is correct.

[0313] Synthesis of Compound A23-4

[0314] A23-3 (2 g, 7.32 mmol) and A23-SM3 (3.28 g, 14.63 mmol) were added to a 50 mL single-mouth bottle, and DMF (20 mL) was added. Potassium bicarbonate (2.2 g, 21.96 mmol) was added, and the temperature was raised to 70 ° C for reaction. The reaction was stopped after 24 hours, and the reaction solution was filtered. The filtrate was added with 5% acetic acid aqueous solution, extracted with DCM three times, and PE and EA were added to slurry until the product was a white solid. After concentration, acetonitrile was added to slurry, and the filter cake was filtered and dried to obtain compound A23-4 (1.1 g, white solid). Yield: 31.6%. LCMS (ESI) m / z calcd. For C 22 H 32 N4O4[M+H] + 417.2; found417.2. It can be seen that the product structure is correct.

[0315] Synthesis of Compound A23-5

[0316] Compound A23-4 (1 g, 2.4 mmol) and DCM (30 mL) were added to a 100 mL single-mouth bottle, followed by 4N HCl in dioxane (8 mL). The mixture was reacted at 25°C for 2 hours under nitrogen protection. After the reaction was completed, the mixture was concentrated to dryness to obtain a crude compound A23-5 (680 mg, yellow solid). Yield: 89.5%. LCMS (ESI) m / z calcd. for C 17 H 24 N4O2[M+H] + 317.2; found 317.3. It can be seen that the product structure is correct.

[0317] Synthesis of Compound A23

[0318] A23-5 (200 mg, 0.632 mmol) and A17-4 (300 mg, 0.632 mmol) were added to a 50 mL single-mouth bottle, and DMF (5 mL) was added. After adding potassium bicarbonate (210 mg, 1.89 mmol), the temperature was raised to 70°C for reaction. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 10:1). The reaction was stopped after 16 hours. The reaction solution was filtered and the filtrate was concentrated. After column chromatography, the eluent ratio was DCM: MeOH = 100:1 to 10:1 to obtain 150 mg of a yellow solid. Compound A23 (71 mg, off-white solid, 96.1%) was obtained by TLC plate. Yield: 14.0%. LCMS (ESI) m / z calcd. For C 41 H 43 F3N8O4[M+H] + 769.3; found 769.4. It can be seen that the product structure is correct.

[0319] Embodiment 24

[0320] In this example, compound 3-[({3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxyphenyl}carbonyl)amino]-5-(trifluoromethyl)benzene-1-carboxamide (Compound A24) was synthesized.

[0321]

[0322] Synthesis of Compound A24-1

[0323] A15-5 (1.5 g, 3.32 mmol) was added to a 25 mL single-mouth bottle, DMF (15 mL), TEA (1 g, 3.32 mmol), HATU (1.3 g, 3.32 mmol), stirred at room temperature for 10 min, 3-amino-5-trifluoromethylbenzamide (812 mg, 3.98 mmol) was added, the temperature was raised to 30 ° C for 18 h, and then the reaction was stopped. The reaction solution was poured into 30 mL of water, filtered to obtain a filter cake, dried, and 10 mL of methanol was added to pulp and filter to obtain a filter cake. Drying gave compound A24-1 (1.1 g, off-white solid). Yield: 52.0%. LCMS (ESI) m / z calcd.For C 32 H 33 F3N6O5[M+H] + 639.3; found 639.47. It can be seen that the product structure is correct.

[0324] Synthesis of Compound A24-2

[0325] A24-1 (600 mg, 0.94 mmol) and dichloromethane (20 mL) were added to a 50 mL single-mouth bottle, and HCl in 1,4-dioxane (4 M, 4 mL) was added dropwise. The reaction was stopped after 4 h at room temperature. A large amount of solid precipitated, which was filtered (rinsed with DCM), and the filter cake was dried to obtain compound A24-2 (550 mg, yellow solid). Yield: 94.0%.

[0326] Synthesis of Compound A24

[0327] A24-2 (300 mg, 0.52 mmol) and A15-SM2 (270 mg, 0.68 mmol) were added to a 50 mL single-mouth bottle, acetonitrile (15 mL), potassium carbonate (250 mg, 1.57 mmol), and the temperature was raised to 60°C for 24 h before the reaction was stopped. The reaction solution was filtered and the filtrate was concentrated and subjected to column chromatography with an eluent ratio of DCM:MeOH = 100:1 to 10:1 to obtain compound A24 (50 mg, off-white solid, 92.7%). Yield: 11.2%. LCMS (ESI) m / z calcd.For C 43 H 40 F3N9O7[M+H] + 852.3; found852.5. 1H NMR (400MHz, DMSO-d6) δ11.00(s,1H),10.65(s,1H),10.60(s,1H),8.56(s,1H),8.46(d,J=16.4Hz,3H),8.25(s,1H),8.20(d,J=2.3Hz ,1H),8.07(dd,J=8.7,2.3Hz,1H),8.01(s,1H),7.96(s,1H),7.75–7.65(m,2H),7.62(d,J=8.2Hz,2H),7.27(d,J=8.9Hz,1H),5.09(dd, J=13.3,5.1Hz,1H),4.44(d,J=17.3Hz,1H),4.30(d,J=17.3Hz,1H),3.95(s,3H),3.77(s,1H),3.17(s,1H),2.97(d,J=7.5Hz,3H),2.71(s,1H),2.60(d,J=19.5Hz,3H),2.45–2.32(m,1H),2.14(s,2H),2.00(t,J=8.2Hz,1H),1.88(d,J=12.1Hz,2H),1.58(t,J=12.1Hz,2H). It can be seen that the product structure is correct.

[0328] Embodiment 25

[0329] In this example, compound 3-[({3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxyphenyl}carbonyl)(methyl)amino]-5-(trifluoromethyl)benzene-1-carboxamide (Compound A25) was synthesized.

[0330]

[0331] Synthesis of Compound A25-1

[0332] A9-SM1 (1 g, 3.62 mmol) was added to a 100 mL three-necked flask, and then oxalyl chloride (1.1 g, 10.87 mmol) and DMF (0.1 mL) were added, and the reaction solution was stirred at room temperature for 2 h. The reaction solution was spin-dried and DCM was dragged 3 times to obtain a light yellow solid. After DCM (20 mL) was added to the above crude product, TEA (300 mg, 7.24 mmol) was added, and the reaction solution was cooled to 0 ° C. A25-SM1 (952 mg, 4.35 mmol) was slowly added dropwise. After the dropwise addition, the reaction solution was warmed to room temperature for 2 hours. After the reaction was completed, DCM was extracted, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by silica gel column chromatography (PE / EA=100 / 1~20 / 1) to obtain compound A25-1 (950 mg, light yellow solid). The yield was 54.9%. LCMS (ESI) m / z calcd.for C 17 H 13 F3INO4[M+H] + 480.0; found 480.1. It can be seen that the product structure is correct.

[0333] Synthesis of Compound A25-2

[0334] Compound A25-1 (800 mg, 0.0295 mol) and DMF (20 mL) were added to a 100 mL single-mouth bottle in sequence, and iodomethane (12.56 g, 88.5 mmol) was added dropwise at 0 °C. The mixture was reacted at room temperature for 4 hours under nitrogen protection. After the reaction was completed, the reaction solution was warmed to room temperature, poured into water (30 mL), extracted three times with EA (30 mL), and the organic phase was concentrated to dryness. The crude product was purified by silica gel column chromatography (PE / EA=100 / 1-20 / 1) to obtain compound A25-2 (800 mg, yellow oil, purity: 80.1%). Yield: 77.81%. LCMS (ESI) m / z calcd.for C 18 H 15 F3INO4[M+H] + 494.0 found 494.1. It can be seen that the product structure is correct.

[0335] Synthesis of Compound A25-3

[0336] In a 100mL single-mouth bottle, A25-2 (800mg, 1.63mmol) was dissolved in MeOH (10mL) and THF (10mL), and NaOH (3M) in H2O (10mL) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for reaction. The reaction was stopped after 16h. The reaction solution was concentrated to remove most of the methanol and water, and the pH was adjusted to 4 with 1M HCl. A large amount of white solid precipitated, which was filtered and dried. Compound A25-3 (700mg, white solid) was obtained. Yield: 90.1%.

[0337] Synthesis of Compound A25-4

[0338] A25-3 (600 mg, 1.26 mmol) was dissolved in DCM (20 mL) in a 100 mL single-mouth bottle. After adding oxalyl chloride (380 mg, 3.77 mmol) and DMF (0.1 mL), the reaction solution was stirred at room temperature for 2 h, the reaction solution was spin-dried, and DCM was dragged 3 times to obtain a light yellow solid. After THF (20 mL) was added to the above crude product, the reaction solution was cooled to 0 ° C. Slowly drop (550 mg, 3.77 mmol) of THF (10 mL) solution. After the drop was completed, the reaction solution was warmed to room temperature and reacted for 2 h. After the reaction was completed, it was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by silica gel column chromatography (PE / EA=100 / 1~2 / 1) to obtain compound A25-4 (350 mg, light yellow solid). Yield 58.4%. LCMS (ESI) m / z calcd.for C 17 H 14 F3IN2O3[M+H] + 479.0 found 479.1. It can be seen that the product structure is correct.

[0339] Synthesis of Compound A25-5

[0340] Compound A25-4 (350 mg, 0.735 mmol), A25-SM2 (117 mg, 0.882 mmol), CuI (30 mg, 0.148 mmol), Pd(PPh3)2Cl2 (52 mg, 0.0735 mmol), DIEA (630 mg, 5.145 mmol) and 1,4-dioxane (10 mL) were added to a 50 mL single-mouth bottle in sequence. The mixture was reacted at 80 °C for 4 hours under nitrogen protection. After the reaction was completed, the mixture was poured into water (1 mL) to precipitate a solid, which was then prepared by a plate (DCM: MeOH = 30: 1) to obtain compound A25-5 (152 mg, white solid, purity 91.0%). Yield: 43.7%. LCMS (ESI) m / z calcd. for C 23 H 16ClF3N4O3[M+H] + 489.1; found 489.2. It can be seen that the product structure is correct.

[0341] Synthesis of Compound A25-6

[0342] A25-5 (1 g, 2.05 mmol) was dissolved in isopropanol (30 mL) in a 50 mL single-mouth bottle, and A25-SM3 (1.1 g, 2.45 mmol) and DIEA (1.2 g, 4.11 mmol) were added. The temperature was raised to 85 ° C. After reacting for 18 h, the reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 25: 1). The reaction was stopped when the reaction was completed. The solvent was concentrated, 20 mL of water and DCM (30 mL x 3) were added for extraction, the organic phases were combined, anhydrous sodium sulfate was added for drying, and the crude product was spin-dried and subjected to column chromatography (eluent ratio PE: EA = 100: 1 to 2: 1) to obtain compound A25-6 (850 mg, beige solid). Yield: 56.1%. LCMS (ESI) m / z calcd.forC 33 H 35 F3N6O5[M+H] + 653.3; found 653.4. It can be seen that the product structure is correct.

[0343] Synthesis of Compound A25-7

[0344] A25-6 (600 mg, 0.94 mmol) and dichloromethane (20 mL) were added to a 50 mL single-mouth bottle, and HCl in 1,4-dioxane (4 M, 4 mL) was added dropwise. The reaction was stopped after 4 h. A large amount of solid precipitated, which was filtered (rinsed with DCM) and the filter cake was dried to obtain compound A25-7 (550 mg, yellow solid). Yield: 94.0%. LCMS (ESI) m / z calcd.for C 28 H 27 F3N6O3[M+H] + 553.2; found 553.2. It can be seen that the product structure is correct.

[0345] Synthesis of Compound A25

[0346] A25-7 (300 mg, 0.52 mmol) and A15-SM2 (270 mg, 0.68 mmol) were added to a 50 mL single-mouth bottle, and DMF (15 mL) was added. Potassium bicarbonate (250 mg, 1.57 mmol) was added, and the temperature was raised to 70°C for 24 h to stop the reaction. The reaction solution was filtered, and the filtrate was concentrated and subjected to column chromatography with an eluent ratio of DCM:MeOH = 100:1 to 10:1 to obtain compound A25 (53 mg, off-white solid, 92.1%). Yield: 11.7%. LCMS (ESI) m / z calcd.for C 44 H 42 F3N9O7[M+H] + 866.3; found866.3. It can be seen that the product structure is correct.

[0347] Embodiment 26

[0348] In this example, the compound N-{5-[(dimethylamino)methyl]-3-(trifluoromethyl)phenyl}-3-({2-[(2-{4-[4-(2,6-dioxypyridin-3-yl)phenyl]pyridin-1-yl}ethyl)amino]pyrimidin-5-yl}ethynyl)-4-methoxybenzamide (Compound A26) was synthesized.

[0349]

[0350] Synthesis of Compound A26-1

[0351] A26-SM1 (10 g, 37.16 mmol), A26-SM2 (17 g, 55.74 mmol), 1,4-dioxane (100 mL) and water (20 mL) were added to a 500 mL single-mouth bottle, potassium carbonate (10.3 g, 74.32 mmol), Pd(dppf)Cl2 (1.4 g, 1.86 mmol), and the temperature was raised to 90°C for reaction after argon replacement. The reaction was stopped after 7 hours, the solvent was added with water, extracted with ethyl acetate three times, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, the residue was slurried with PE:EA=2:1 (50 mL), filtered to obtain a filter cake, and dried to obtain compound A26-1 (7.1 g, off-white solid). Yield: 47.7%.

[0352] Synthesis of Compound A26-2

[0353] A26-1 (7.1 g, 20.73 mmol) was added to a 500 mL single-mouth bottle, dissolved in EtOH (150 mL), and Pd / C (0.3 g) was added. The reaction solution was stirred overnight at room temperature in a hydrogen atmosphere. TLC (PE / EA=0 / 1) confirmed the completion of the reaction. After the reaction was completed, the reaction solution was filtered through diatomaceous earth and the filtrate was dried. The crude product was purified by silica gel column chromatography (DCM / MeOH=100 / 1~10 / 1) to obtain compound A26-2 (5.2 g, gray solid, purity: 90.3%). Yield: 64.4%. LCMS (ESI) m / z calcd.forC 21 H 28 N2O4[M+H] + 373.2; found 373.3. It can be seen that the product structure is correct.

[0354] Synthesis of Compound A26-3

[0355] Compound A26-2 (3 g, 5.79 mmol) and DCM (30 mL) were added to a 100 mL single-mouth bottle, followed by 4N HCl in dioxane (8 mL). The mixture was reacted at 25 °C for 2 hours under nitrogen protection. After the reaction was completed, the mixture was concentrated to dryness to obtain a crude compound A26-3 (2.1 g, yellow solid). Yield: 86.2%. LCMS (ESI) m / z calcd. for C 16 H 20 N2O2[M+H] + 273.2; found 273.2. It can be seen that the product structure is correct.

[0356] Synthesis of Compound A26-4

[0357] A26-3 (2 g, 7.32 mmol) and A23-SM3 (3.28 g, 14.63 mmol) were added to a 50 mL single-mouth bottle, and DMF (20 mL) was added to dissolve. After adding potassium bicarbonate (2.2 g, 21.96 mmol), the solution was in a suspension state. The temperature was raised to 70 °C for reaction. TLC monitored the progress of the reaction (developing solvent: DCM: MeOH = 10:1), and the reaction was stopped after 24 hours. The reaction solution was filtered, and the filtrate was added with 5% acetic acid aqueous solution, extracted with DCM three times, PE and EA were added to pulp until the product was a white solid, and acetonitrile was added to pulp after rotary evaporation, and the filter cake was filtered and dried to obtain compound A26-4 (1.1 g, white solid). Yield: 31.6%. LCMS (ESI) m / z calcd.ForC 23 H 33 N3O4[M+H] + 416.3; found 416.3. It can be seen that the product structure is correct.

[0358] Synthesis of Compound A26-5

[0359] Compound A26-4 (1 g, 2.4 mmol) and DCM (30 mL) were added to a 100 mL single-mouth bottle, followed by 4N HCl in dioxane (8 mL). The mixture was reacted at 25 °C for 2 hours under nitrogen protection. After the reaction was completed, the mixture was concentrated to dryness to obtain a crude compound A26-5 (680 mg, yellow solid). Yield: 89.5%. LCMS (ESI) m / z calcd. for C 18 H 25 N3O2[M+H] + 316.2; found 316.3. It can be seen that the product structure is correct.

[0360] Synthesis of Compound A26-6

[0361] A26-5 (200 mg, 0.632 mmol) and A17-4 (300 mg, 0.632 mmol) were added to a 50 mL single-mouth bottle, and DMF (5 mL) was added to partially dissolve. After adding potassium bicarbonate (210 mg, 1.89 mmol), the temperature was raised to 70 ° C for reaction. TLC monitored the reaction progress (developing solvent: DCM: MeOH = 10: 1), and the reaction was stopped after 16 hours. The reaction solution was filtered and the filtrate was concentrated. After column chromatography, the eluent ratio was DCM: MeOH = 100: 1 to 10: 1 to obtain 200 mg of yellow solid. Compound A26 (51 mg, off-white solid, 96.1%) was obtained by preparation plate. Yield: 11.7%. LCMS (ESI) m / z calcd. For C 42 H 44 F3N7O4[M+H] + 768.3; found768.4. It can be seen that the product structure is correct.

[0362] Embodiment 27

[0363] In this example, compound 3-({[3-({2-[(2-{4-[4-(2,4-dioxypyrimidin-1-yl)phenyl]pyrimidin-1-yl}ethyl)amino]pyrimidin-5-yl}ethynyl)-4-methoxyphenyl]carbonyl}amino)-5-(trifluoromethyl)benzene-1-carboxamide (Compound A27) was synthesized.

[0364]

[0365] A23-5 (200 mg, 0.632 mmol) and A18-2 (280 mg, 0.632 mmol) were added to a 50 mL single-mouth bottle, and DMF (5 mL) was added. Potassium bicarbonate (210 mg, 1.89 mmol) was added, and the temperature was raised to 70°C for 16 h before the reaction was stopped. The reaction solution was filtered, and the filtrate was concentrated and subjected to column chromatography with an eluent ratio of DCM:MeOH = 100:1 to 10:1 to obtain compound A27 (61 mg, off-white solid, 96.1%). Yield: 13.2%. LCMS (ESI) m / z calcd.For C 39 H 37 F3N8O5[M+H] + 755.3; found755.4. It can be seen that the product structure is correct.

[0366] Embodiment 28

[0367] In this example, compound 3-({[3-({2-[(2-{4-[4-(2,6-dioxypyridin-3-yl)phenyl]pyridin-1-yl}ethyl)amino]pyrimidin-5-yl}ethynyl)-4-methoxyphenyl]carbonyl}amino)-5-(trifluoromethyl)benzene-1-carboxamide (Compound A28) was synthesized.

[0368]

[0369] A26-5 (200 mg, 0.632 mmol) and A18-2 (300 mg, 0.632 mmol) were added to a 50 mL single-mouth bottle, and DMF (5 mL) was added. Potassium bicarbonate (210 mg, 1.89 mmol) was added, and the temperature was raised to 70°C for 16 h. The reaction was stopped, and the reaction solution was filtered. The filtrate was concentrated and subjected to column chromatography. The eluent ratio was DCM:MeOH = 100:1 to 10:1 to obtain 200 mg of a yellow solid. Compound A28 (41 mg, off-white solid, 97.1%) was obtained by preparation. Yield: 13.2%. LCMS (ESI) m / z calcd.ForC 40 H 38 F3N7O5[M+H] + 754.3; found 754.4. It can be seen that the product structure is correct.

[0370] Embodiment 29

[0371] In this example, compound 3-[({3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]oxy}propyl)pyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxyphenyl}carbonyl)amino]-5-(trifluoromethyl)benzene-1-carboxamide (Compound A29) was synthesized.

[0372]

[0373] A20-2 (90 mg, 0.280 mmol) and A24-2 (150 mg, 0.280 mmol) were dissolved in DMF (5 ml), and AcOH (34 mg, 0.56 mmol) and STAB (120 mg, 0.56 mmol) were added. The mixture was reacted for 24 h under argon protection. In an ice-water bath, the reaction solution was poured into a beaker containing a saturated NaHCO3 solution, stirred for 10 mins, extracted with DCM, dried and concentrated to obtain a crude product. Column chromatography purification (DCM: MeOH = 150: 1 to 10: 1) gave compound A29 (55 mg, off-white solid, purity: 96.8%). Yield: 16.1%. LCMS (ESI) m / z calcd. for C 43 H 41 F3N8O7[M+H] + 839.3; found 839.4. It can be seen that the product structure is correct.

[0374] Embodiment 30

[0375] In this example, compound 3-[({3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-4-yl]oxy}propyl)pyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxyphenyl}carbonyl)amino]-5-(trifluoromethyl)benzene-1-carboxamide (Compound A30) was synthesized.

[0376]

[0377] A21-2 (100 mg, 0.316 mmol) and A24-2 (170 mg, 0.316 mmol) were dissolved in DMF (5 ml), and AcOH (38 mg, 0.632 mmol) and STAB (135 mg, 0.632 mmol) were added, and the mixture was reacted for 24 h under argon protection. In an ice-water bath, the reaction solution was poured into a beaker containing a saturated NaHCO3 solution, stirred for 10 mins, extracted with DCM, dried and concentrated to obtain a crude product. Column chromatography purification (DCM: MeOH = 150: 1 to 10: 1) gave compound A30 (30 mg, off-white solid, purity: 96.2%). Yield: 10.3%. LCMS (ESI) m / z calcd. for C 43 H 41 F3N8O7[M+H] + 839.3; found 839.4. It can be seen that the product structure is correct.

[0378] Embodiment 31

[0379] In this example, compound 3-[({3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxyphenyl}carbonyl)amino]-5-methoxybenzene-1-carboxamide (Compound A31) was synthesized.

[0380]

[0381] Synthesis of Compound A31-11

[0382] A31-SM (10 g, 42.52 mmol) was added to a 500 mL single-mouth bottle. After adding oxalyl chloride (26 g, 212.60 mmol) and DMF (0.5 mL), the reaction solution was stirred at room temperature for 2 h, the reaction solution was spin-dried, and DCM was dragged 3 times to obtain a light yellow solid. After adding DCM (100 mL) to the above crude product, the reaction solution was cooled to 0 ° C, and a solution of NH4OH (3 g, 86.4 mmol) in DCM (20 mL) was slowly added dropwise. After the dropwise addition, the reaction solution was warmed to room temperature and the reaction was continued for 2 hours. Wash with brine, dry with anhydrous sodium sulfate, filter and spin-dry. The crude product was purified by silica gel column chromatography (PE / EA=100 / 1~1 / 1) to obtain compound A31-11 (5.7 g, light yellow solid). Yield 58.4%. LCMS (ESI) m / z calcd.for C8H8N2O4[M+H] + 197.1; found 197.1. It can be seen that the product structure is correct.

[0383] Synthesis of Compound A31-12

[0384] A31-11 (5.5 g) was added to a 500 mL single-mouth bottle, dissolved in EtOH (110 mL), and Pd / C (0.3 g) was added. The reaction solution was stirred overnight at room temperature in a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered through diatomaceous earth and the filtrate was dried by spin drying. The crude product was purified by silica gel column chromatography (PE / EA=100 / 1~0 / 1) to obtain A31-12 (4.1 g, yellow solid). The yield was 85.8%. LCMS (ESI) m / z calcd.for C8H 10 N2O2[M+H] + 167.1; found 167.2. It can be seen that the product structure is correct.

[0385]

[0386] Synthesis of Compound A31-1

[0387] A16-3 (1.5 g, 3.31 mmol) was added to a 25 mL single-mouth bottle, and DMF (15 mL) was added. The solution was white and turbid. TEA (2 g, 3.62 mmol) and HATU (1.3 g, 3.31 mmol) were then added. The solution was still turbid. A31-12 (825 mg, 4.97 mmol) was added, and the temperature was raised to 30 ° C. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 15: 1). The reaction was stopped after 18 hours. The reaction solution was poured into 30 mL of water, filtered to obtain a filter cake, dried, and 10 mL of methanol was added to slurry and filter to obtain a filter cake. Compound A31-1 (1.1 g, off-white solid) was obtained by drying. Yield: 55.2%. LCMS (ESI) m / z calcd. For C 32 H 36 N6O6[M+H] + 601.3; found 601.4. It can be seen that the product structure is correct.

[0388] Synthesis of Compound A31-2

[0389] A31-1 (600 mg, 0.998 mmol) was added to a 50 mL single-mouth bottle. Dichloromethane (20 mL) was added. The solution was partially undissolved. HCl in 1,4-dioxane (4 M, 4 mL) was added dropwise. During the addition, the solution changed from turbid to clear, and then from clear to turbid. TLC was used to monitor the reaction progress (developing solvent: DCM: MeOH = 15:1). The reaction was stopped after 4 hours. A large amount of solid precipitated in the reaction system. It was filtered (the filter cake was rinsed with DCM). The filter cake was dried to obtain compound A31-2 (480 mg, yellow solid). Yield: 91.0%.

[0390] Synthesis of compound A31

[0391] A31-2 (300 mg, 0.599 mmol) and A15-SM2 (270 mg, 0.899 mmol) were added to a 50 mL single-mouth bottle, and acetonitrile (15 mL) was added to partially dissolve. After adding potassium carbonate (250 mg, 1.797 mmol), the solution was still in a suspension state. The temperature was raised to 60 ° C for reaction. TLC monitored the progress of the reaction (developing solvent: DCM: MeOH = 10: 1), and the reaction was stopped after 24 hours. The reaction solution was filtered, and the filtrate was concentrated. After column chromatography, the eluent ratio was DCM: MeOH = 100: 1 to 10: 1, and 150 mg of yellow solid was obtained. Compound A31 (38 mg, off-white solid, 96.5%) was obtained by preparation plate. Yield: 7.52%. LCMS (ESI) m / z calcd. For C 43 H 43 N9O8[M+H] + 814.3; found 814.4. It can be seen that the product structure is correct.

[0392] Embodiment 32

[0393] In this example, compound 3-({2-[(3-{4-[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yl}propionyl)amino]pyrazolo[1,5-a]pyrimidin-6-yl}ethynyl)-4-ethyl-N-(3-methoxyphenyl)benzamide (Compound A32) was synthesized.

[0394]

[0395] Synthesis of Compound A32-11

[0396] At room temperature, A32-SM1 (5 g, 15.47 mmol) and A32-SM2 (3 g, 15.47 mmol) were dissolved in 1,4-dioxane (50 mL) and C SCO3 (3.58 g, 18.57 mmol), and then Pd-PEPPSI-IHeptCl (752 mg, 0.78 mmol) was added. Under argon protection, the temperature was raised to 100°C for 18 h to stop the reaction. Saturated NaCl aqueous solution was added, and DCM (50 mL × 3) was used for extraction. The organic phases were combined, dried and concentrated, and purified by column chromatography (DCM: MeOH = 100: 1 / 20: 1) to obtain compound A32-11 (4.2 g, light yellow solid, purity 92.43%). Yield: 58.55%. LCMS (ESI) m / z calcd. For C 22 H 28 N4O5[M+H] + 429.2; found 429.2. It can be seen that the product structure is correct.

[0397] Synthesis of Compound A32-12

[0398] Under magnetic stirring, in a 100mL single-mouth bottle, A32-11 (4g, 9.34mmol) was dissolved in MeOH (40mL), and HCl in 1,4-dioxane (10mL, 4M) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for 4h to stop the reaction. After the reaction was completed, the mixture was filtered and dried. Compound A32-12 (3.4g, light yellow solid, purity 88.67%) was obtained. Yield: 88.52%.

[0399] Synthesis of Compound A32-13

[0400] A32-12 (2 g, 5.48 mmol) was dissolved in MeCN (50 mL), tert-butyl 3-bromopropionate (1.3 g, 6.02 mmol) and DIEA (2.2 g, 16.44 mmol) were added, and the mixture was reacted in an oil bath at 60°C. The reaction was stopped after 16 h. The reaction solution was concentrated and purified by column chromatography. Compound A32-13 (2.2 g, off-white solid, purity 93.2%) was obtained. Yield: 81.9%. LCMS (ESI) m / z calcd. For C 24 H 32 N4O5[M+H] + 457.2; found 457.3. It can be seen that the product structure is correct.

[0401] Synthesis of Compound A32-14

[0402] A32-13 (2 g, 4.82 mmol) was dissolved in DCM (30 mL), TFA (5 mL) was added, and the mixture was reacted at room temperature for 5 h. The reaction solution was concentrated to dryness to obtain compound A32-14 (1.9 g, yellow oil, purity 91.7%). Yield: 90.4%.

[0403] Synthesis of Compound A32-21

[0404] A32-SM3 (20 g, 204 mmol) was dissolved in ethanol (200 mL), A32-SM4 (30 g, 204 mmol) and p-toluenesulfonic acid (7 g, 40.8 mmol) were added, and the mixture was heated to 80 °C for 5 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, 100 mL of ethyl acetate was added, and the mixture was washed with saturated sodium bicarbonate solution (50 mL) and saturated sodium chloride solution (50 mL) in sequence, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (PE: EA = 50: 1 to 3: 1) to obtain compound A32-21 (22 g, yellow solid, purity 91.32%). Yield: 51.2%. LCMS (ESI) m / z calcd. For C6H5BrN4 [M+H] + 213.0; found 213.1. It can be seen that the product structure is correct.

[0405] Synthesis of Compound A32-22

[0406] Compound A32-21 (10 g, 46.94 mmol), trimethylethynylsilane (6.92 g, 70.41 mmol), CuI (900 mg, 4.69 mmol), Pd(PPh3)2Cl2 (1 g, 2.35 mol), DIEA (18 g, 140.82 mmol) and DMF (5 mL) were added to a 500 mL single-mouth bottle in sequence. The reaction was carried out at 100 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (50 mL) and extracted three times with EA (50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A32-22 (7 g, brown oil, purity 90.22%) was obtained. Yield: 58.41% LCMS (ESI) m / z calcd.for C 11 H 14 N4Si[M+H] + 231.1 found 231.3. It can be seen that the product structure is correct.

[0407] Synthesis of Compound A32-23

[0408] At 0°C, A32-22 (7 g, 30.39 mmol) was dissolved in MeOH (70 mL), K2CO3 (8.4 g, 60.78 mmol) was added and stirred for 5 min, then the mixture was moved to room temperature for 1 h. The reaction solution was filtered and concentrated to dryness, and the residue was purified by silica gel chromatography with an eluent and a ratio of PE / EtOAc = 20 / 1 to 5 / 1 to obtain compound A32-23 (4.4 g, yellow solid, purity 91.21%). Yield: 83.50%. LCMS (ESI) m / z calcd. For C8H6N4 [M+H] + 159.1; found 159.1. It can be seen that the product structure is correct.

[0409]

[0410] Synthesis of compound A32-1

[0411] A2-SM (500 mg, 1.81 mmol) was dissolved in DMF (8 mL), DIEA (700 mg, 5.43 mmol) and HATU (690 mg, 1.81 mmol) were added, and after stirring for 5 min, 3-methoxyaniline (335 mg, 2.72 mmol) dissolved in DMF (2 mL) was added, and the reaction was allowed to react at room temperature for 16 h. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 10: 1). After stopping the reaction, H2O was added to the reaction solution, and extracted with EA (20 mL x 3). The organic phase was concentrated and purified and separated by column chromatography (PE: EA = 10: 1). Compound A32-1 (530 mg, white solid, purity 92.33%) was obtained. Yield: 70.88%. LCMS (ESI) m / z calcd. for C 16 H 16 INO2[M+H] + 382.0; found382.1. It can be seen that the product structure is correct.

[0412] Synthesis of Compound A32-2

[0413] Compound A32-1 (500 mg, 1.31 mmol), A32-23 (312 mg, 1.97 mmol), CuI (50 mg, 0.262 mmol), Pd(PPh3)2Cl2 (10 mg, 0.131 mol), DIEA (510 mg, 3.93 mmol) and DMF (5 mL) were added to a 100 mL single-mouth bottle in sequence. The reaction was carried out at 100 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (150 mL) and extracted three times with EA (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A32-2 (330 mg, brown oil, purity 88.73%) was obtained. Yield: 54.26%. LCMS (ESI) m / z calcd.for C 24 H 21 N5O2[M+H] + 412.1; found 412.2. It can be seen that the product structure is correct.

[0414] Synthesis of compound A32

[0415] A32-2 (200 mg, 0.486 mmol) was dissolved in DMF (4 mL), TEA (150 mg, 1.45 mmol) and HATU (185 mg, 0.486 mmol) were added, and after stirring for 5 min, A32-14 (234 mg, 0.583 mmol) dissolved in DMF (1 mL) was added and reacted at room temperature. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 10: 1), and the reaction was stopped after 20 h. H2O was added to the reaction solution, and it was extracted with EA (15 mL x 3), and the organic phase was concentrated and purified and separated by preparing plates for multiple times (DCM: MeOH = 10: 1). Compound A32 (78 mg, white solid, purity 96.22%) was obtained. Yield: 19.45%. LCMS (ESI) m / z calcd.for C 44 H 43 N9O6[M+H] + 794.3; found 794.4. It can be seen that the product structure is correct.

[0416] Embodiment 33

[0417] In this example, compound 3-({2-[4-({1-[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]pyridin-4-yl}methyl)piperazine-1-yl]pyrazolo[1,5-a]pyrimidin-6-yl}ethynyl)-4-ethyl-N-(3-methoxyphenyl)benzamide (Compound A33) was synthesized.

[0418]

[0419] Synthesis of Compound A33-1

[0420] At room temperature, A33-SM1 (2 g, 7.06 mmol) and A33-SM2 (2.3 g, 7.06 mmol) were dissolved in 1,4-dioxane (40 mL) and C S CO3 (4.6 g, 14.12 mmol), and then Pd-PEPPSI-IHeptCl (700 mg, 0.706 mmol) was added. Under argon protection, the temperature was raised to 100°C for 18 h to stop the reaction. Saturated NaCl and DCM (30 mL×3) were used for extraction, and the organic phases were combined, dried and concentrated, and purified by column chromatography (DCM: MeOH = 100: 1 / 30: 1) to obtain compound A33-1 (1.9 g, light yellow solid, purity 92.32%). Yield: 47.3%. LCMS (ESI) m / z calcd.For C 28 H 39 N5O5[M+H] + 526.3; found 526.4. It can be seen that the product structure is correct.

[0421] Synthesis of Compound A33-2

[0422] In a 250 mL single-mouth bottle, A33-1 (1.9 g, 3.61 mmol) was dissolved in DCM (40 mL), and HCl in 1,4-dioxane (10 mL, 4 M) was slowly added dropwise under ice bath conditions. The reaction was stopped after 4 h at room temperature. After the reaction was completed, the mixture was filtered and dried to obtain compound A33-2 (1.4 g, light yellow solid, purity 92.61%). Yield: 77.6%.

[0423] Synthesis of compound A33-3

[0424] Compound A32-1 (1.4 g, 3.67 mmol), A34-12 (890 mg, 5.51 mmol), CuI (100 mg, 0.734 mmol), Pd(PPh3)2Cl2 (260 mg, 0.367 mol), DIEA (1.4 g, 11.01 mmol) and DMF (20 mL) were added to a 100 mL single-mouth bottle in sequence. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (50 mL) and extracted three times with EA (50 mL). Dry, filter, and concentrate to dryness. Compound A33-3 (1.5 g, brown foam product, purity 90.22%) was obtained by column chromatography purification and separation (DCM: MeOH = 20: 1). Yield: 50.55%. LCMS (ESI) m / z calcd.for C 24 H 19 FN4O2[M+H] + 415.2; found 415.2. It can be seen that the product structure is correct.

[0425] Synthesis of compound A33

[0426] A33-3 (200 mg, 0.501 mmol) and A33-2 (250 mg, 0.551 mmol) were dissolved in DMF (5 mL), K2CO3 (208 mg, 1.51 mmol) was added, and the reaction was stopped at 90°C for 16 h. H2O was added to the reaction solution, and it was extracted with DCM (20 mL x 3). The organic phase was concentrated and purified by column chromatography (DCM: MeOH = 8: 1) to obtain compound A33 (71 mg, white solid, purity 97.22%). Yield: 16.81%. LCMS (ESI) m / z calcd. for C 47 H 49 N9O5[M+H] + 820.4; found820.4. It can be seen that the product structure is correct.

[0427] Embodiment 34

[0428] In this example, the compound N-[3-(dimethylamino)phenyl]-3-[(2-{[1-({1-[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]pyridin-4-yl}carbonyl)pyridin-4-yl]oxy}pyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]-4-ethylbenzamide (Compound A34) was synthesized.

[0429]

[0430] Synthesis of Compound A34-11

[0431] Compound A34-SM1 (5 g, 23.15 mmol), trimethylethynylsilane (3.41 g, 34.72 mmol), CuI (900 mg, 4.63 mmol), Pd(PPh3)2Cl2 (1 g, 2.31 mmol), DIEA (9 g, 69.45 mmol) and DMF (50 mL) were added to a 250 mL single-mouth bottle in sequence. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (50 mL) and extracted three times with EA (50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A34-11 (4.4 g, brown oil, purity 80.17%) was obtained. Yield: 65.32%. LCMS (ESI) m / z calcd.for C 11 H 12 FN3Si[M+H] + 234.1; found 234.1. It can be seen that the product structure is correct.

[0432] Synthesis of Compound A34-12

[0433] At 0°C, A34-11 (4.4 g, 18.86 mmol) was dissolved in MeOH (70 mL), K2CO3 (5.21 g, 37.72 mmol) was added and stirred for 5 min, then the mixture was moved to room temperature for 1 h. The reaction solution was filtered and concentrated to dryness, and the residue was purified by silica gel column chromatography with an eluent and a ratio of PE / EtOAc = 20 / 1 to 5 / 1 to obtain compound A34-12 (2.2 g, yellow solid, purity 91.21%). Yield: 66.03%. LCMS (ESI) m / z calcd. For C8H4FN3 [M+H] + 162.0; found 162.1. It can be seen that the product structure is correct.

[0434] Synthesis of Compound A34-13

[0435] A34-12 (2.2 g, 13.65 mmol) and A34-SM2 (3.3 g, 16.38 mmol) were dissolved in DMF (40 mL), K2CO3 (3.8 g, 27.31 mmol) was added, and the mixture was heated to 80°C for 5 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, 50 mL of ethyl acetate was added, washed with saturated sodium chloride solution (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (PE:EA=50:1-20:1) to obtain compound A34-12 (2.8 g, yellow solid, purity 95.43%). Yield: 57.16%. LCMS (ESI) m / z calcd.For C 18 H 22 N4O3[M+H] + 343.2; found 343.2. It can be seen that the product structure is correct.

[0436]

[0437] Synthesis of Compound A34-21

[0438] At room temperature, A33-SM2 (10 g, 30.95 mmol) and tert-butyl 4-piperidincarboxylate (5.7 g, 30.95 mmol) were dissolved in 1,4-dioxane (50 mL), and C S CO3 (11.94 g, 61.89 mmol), and then Pd-PEPPSI-IHeptCl (1.4 g, 1.55 mmol) was added. The argon gas was replaced 3 times, and the temperature was raised to 100°C for 18 h to stop the reaction. Saturated NaCl and DCM (50 mL×3) were used for extraction, and the organic phases were combined, dried and concentrated, and purified by column chromatography (DCM: MeOH = 100: 1 / 20: 1) to obtain compound A34-21 (6.3 g, light yellow solid, purity 89.76%). Yield: 42.74%. LCMS (ESI) m / z calcd.ForC 23 H 29 N3O5[M+H] + 428.2; found 428.2. It can be seen that the product structure is correct.

[0439] Synthesis of Compound A34-22

[0440] Under magnetic stirring, in a 250mL single-mouth bottle, A34-21 (6.3g, 14.74mmol) was dissolved in DCM (60mL), and HCl in 1,4-dioxane (10mL, 4M) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for reaction. The reaction was stopped after 4h. After the reaction was completed, the mixture was filtered and dried. Compound A34-22 (4.4g, light yellow solid, purity 87.61%) was obtained. Yield: 70.43%.

[0441]

[0442] Synthesis of Compound A34-1

[0443] A2-SM (10 g, 36.22 mmol) was dissolved in DMF (50 mL), TEA (11 g, 108.6 mmol) and HATU (13 g, 36.22 mmol) were added, and N,N-dimethyl-m-phenylenediamine (5.9 g, 43.47 mmol) was added after stirring for 5 min. The reaction was stopped after 20 h at 40 °C. H2O was added to the reaction solution, and it was extracted with EA (50 mL x 3). The organic phase was concentrated and purified by column chromatography (DCM: MeOH = 100: 1 / 10: 1). Compound A34-1 (8.1 g, white solid, purity 87.22%) was obtained. Yield: 49.47%. LCMS (ESI) m / z calcd.for C 17 H 19 IN2O[M+H] + 395.0; found 395.1. It can be seen that the product structure is correct.

[0444] Synthesis of Compound A34-2

[0445] Compound A34-1 (8.1 g, 20.55 mmol), A34-13 (8.4 g, 24.65 mmol), CuI (800 mg, 4.11 mmol), Pd(PPh3)2Cl2 (1.2 g, 2.06 mol), DIEA (8 g, 61.65 mmol) and DMF (50 mL) were added to a 250 mL single-mouth bottle in sequence. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (50 mL) and extracted three times with EA (50 mL). Dry, filter, and concentrate to dryness. Compound A34-2 (7 g, brown oil, purity 90.22%) was obtained. Yield: 50.6%. LCMS (ESI) m / z calcd.for C 35 H 40 N6O4[M+H] +609.3; found 609.3. It can be seen that the product structure is correct.

[0446] Synthesis of compound A34-3

[0447] In a 100mL single-mouth bottle, A34-2 (4g, 6.74mmol) was dissolved in DCM (40mL), and the solution was light yellow and turbid. HCl in 1,4-dioxane (10mL, 4M) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for reaction. The reaction progress was monitored by TLC (developing solvent: DCM:MeOH=50:1), and the reaction was stopped after 4h. After the reaction was completed, the mixture was filtered and dried. Compound A34-3 (3.1g, light yellow solid, purity 87.67%) was obtained. Yield: 74.01%. LCMS (ESI) m / z calcd.forC 30 H 32 N6O2[M+H] + 509.3; found 509.3. It can be seen that the product structure is correct.

[0448] Synthesis of compound A34

[0449] A34-3 (100 mg, 0.183 mmol) was dissolved in DMF (4 mL), TEA (56 mg, 0.549 mmol), HATU (70 mg, 0.183 mmol), A34-22 (70 mg, 0.183 mmol) were added, and the reaction was stopped at 40 ° C for 20 h. H2O was added to the reaction solution, and EA (20 mL x 3) was used for extraction. The organic phase was concentrated and purified and separated by preparative plate (DCM: MeOH = 8: 1). Compound A34 (24 mg, white solid, purity 96.31%) was obtained. Yield: 14.62%. LCMS (ESI) m / z calcd.for C 49 H 51 N9O6[M+H] + 862.4; found 862.4. It can be seen that the product structure is correct.

[0450] Embodiment 35

[0451] In this example, the compound N-[3-(dimethylamino)phenyl]-3-({2-[4-(2-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]oxy}ethyl)piperazin-1-yl]pyrazolo[1,5-a]pyrimidin-6-yl}ethynyl)-4-ethylbenzamide (Compound A35) was synthesized.

[0452]

[0453] Synthesis of compound A35-1

[0454] At 0°C, A35-SM2 (10 g, 43.42 mmol) was dissolved in THF (50 mL), A35-SM1 (11.3 g, 43.42 mmol) and triphenylphosphine (12.5 g, 47.76 mmol) were added, stirred evenly, and then DEAD (8.2 g, 47.76 mmol) dissolved in THF (20 mL) was slowly added dropwise. The argon gas was replaced 3 times, the temperature was raised to 20°C and the reaction was carried out for 16 h. The reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 30: 1). After the reaction was completed, the reaction was stopped. Saturated NaCl and DCM (50 mL × 3) were used for extraction, the organic phases were combined, the organic phases were dried and concentrated, and purified by column chromatography (DCM: MeOH = 100: 1 / 30: 1) to obtain compound A35-1 (6.1 g, light yellow solid, purity 92.41%). Yield: 29.73%. LCMS(ESI)m / z calcd.For C 24 H 32 N4O6[M+H] + 473.2; found 473.3. It can be seen that the product structure is correct.

[0455] Synthesis of Compound A35-2

[0456] Under magnetic stirring, in a 250mL single-mouth bottle, A35-1 (6.1g, 14.74mmol) was dissolved in DCM (60mL), and the solution was light yellow and turbid. HCl in 1,4-dioxane (15mL, 4M) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for reaction. TLC was used to monitor the reaction progress (developing solvent: DCM:MeOH=20:1), and the reaction was stopped after 4h. After the reaction was completed, the mixture was filtered and dried. Compound A35-2 (4.7g, light yellow solid, purity 91.61%) was obtained. Yield: 89.04%.

[0457]

[0458] Synthesis of compound A35-3

[0459] Compound A34-1 (300 mg, 0.761 mmol), A34-12 (185 mg, 1.14 mmol), CuI (30 mg, 0.152 mmol), Pd(PPh3)2Cl2 (55 mg, 0.076 mol), DIEA (300 mg, 2.28 mmol) and DMF (8 mL) were added to a 250 mL single-mouth bottle in sequence. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (50 mL) and extracted three times with EA (50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A35-3 (260 mg, brown oil, purity 90.72%) was obtained. Yield: 72.51%. LCMS (ESI) m / z calcd.for C 25 H 22 FN5O[M+H] + 428.2; found 428.2. It can be seen that the product structure is correct.

[0460] Synthesis of compound A35

[0461] A35-3 (200 mg, 0.468 mmol) and A35-2 (515 mg, 0.211 mmol) were dissolved in DMF (5 mL), K2CO3 (195 mg, 1.40 mmol) was added, and the reaction was stopped at 80°C for 16 h. H2O was added to the reaction solution, and extracted with DCM (20 mL x 3). The organic phase was concentrated and purified by column chromatography (DCM: MeOH = 10: 1) to obtain compound A35 (54 mg, white solid, purity 96.11%). Yield: 14.22%. LCMS (ESI) m / z calcd. for C 44 H 45 N9O5[M+H] + 780.3; found 780.4. It can be seen that the product structure is correct.

[0462] Embodiment 36

[0463] In this example, the compound N-[3-(dimethylamino)phenyl]-3-[(2-{[1-(4-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-4-oxypyridin-4-yl]amino}pyrazolo[1,5-a]pyrimidin-6-yl)ethynyl]-4-methoxybenzamide (Compound A36) was synthesized.

[0464]

[0465] Synthesis of Compound A36-1

[0466] A9-SM1 (1 g, 3.60 mmol) was dissolved in DMF (15 mL), DIEA (1.4 g, 10.8 mmol), HATU (1.4 g, 3.60 mmol), and N,N-dimethyl-m-phenylenediamine (735 mg, 5.39 mmol) were added, and the reaction was carried out at room temperature. The reaction was stopped after 20 h. H2O was added to the reaction solution, and the organic phase was concentrated and purified and separated by preparing plates for multiple times (PE:EA=10:1). Compound A36-1 (970 mg, white solid, purity 87.43%) was obtained. Yield: 59.51%. LCMS (ESI) m / z calcd.for C 16 H 17 IN2O2[M+H] + 397.0; found 397.1. It can be seen that the product structure is correct.

[0467] Synthesis of Compound A36-2

[0468] Compound A36-1 (970 mg, 2.45 mmol), A32-23 (580 mg, 3.67 mmol), CuI (100 mg, 0.49 mmol), Pd(PPh3)2Cl2 (177 mg, 0.24 mmol), DIEA (950 mg, 7.35 mmol) and DMF (15 mL) were added to a 100 mL single-mouth bottle in sequence. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (150 mL) and extracted three times with EA (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A36-2 (670 mg, brown oil, purity 92.73%) was obtained. Yield: 59.78%. LCMS (ESI) m / z calcd.for C 25 H 24 N6O[M+H] + 425.2; found 425.2. It can be seen that the product structure is correct.

[0469] Synthesis of compound A36-3

[0470] A36-3 (650 mg, 1.53 mmol) and N-tert-butyloxycarbonyl-4-piperidone (366 mg, 1.84 mmol) were dissolved in THF (20 mL), AcOH (92 mg, 1.53 mmol) was added, and NaB(OAc)3 (650 mg, 3.06 mmol) was added after stirring for 5 min. The reaction was stopped at room temperature for 16 h. Dichloromethane and saturated sodium carbonate were added for extraction, and the organic layer was washed with saturated brine, dried, and spin-dried. The mixture was purified and separated by column chromatography (DCM: MeOH = 30: 1) to obtain compound A36-3 (410 mg, white solid, purity 87.34%). Yield: 38.36%. LCMS (ESI) m / z calcd.for C 34 H 39 N7O4[M+H]+610.3; found610.3. It can be seen that the product structure is correct.

[0471] Synthesis of Compound A36-4

[0472] A36-3 (300 mg, 0.492 mmol) was dissolved in DCM (15 mL), and HCl in 1,4-dioxane (3 mL, 4 M) was added dropwise. The reaction was stopped after 6 h at room temperature. The organic phase was concentrated and dried by spin drying. Compound A36-4 (210 mg, white solid, purity 89.65%) was obtained. Yield: 70.07%. LCMS (ESI) m / z calcd.for C 29 H 31 N7O2[M+H] + 510.2; found 510.3. It can be seen that the product structure is correct.

[0473] Synthesis of Compound A36

[0474] A36-4 (210 mg, 0.384 mmol) and A15-SM2 (450 mg, 1.15 mmol) were added to a 50 mL single-mouth bottle, acetonitrile (15 mL), potassium carbonate (160 mg, 1.15 mmol), and the temperature was raised to 60°C for 24 h before stopping. The reaction solution was filtered, and the filtrate was concentrated and subjected to column chromatography with an eluent ratio of DCM:MeOH = 100:1 to 10:1 to obtain compound A36 (41 mg, off-white solid, 95.11%). Yield: 12.32%. LCMS (ESI) m / z calcd.For C 45 H 46 N 10 O6[M+H] + 823.3; found823.4. It can be seen that the product structure is correct.

[0475] Embodiment 37

[0476] In this example, compound 3-{[2-(4-{[1-(2-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}ethyl)pyridin-4-yl]methyl}piperazin-1-yl)pyrazolo[1,5-a]pyrimidin-6-yl]ethynyl}-4-ethyl-N-[3-(trifluoromethyl)phenyl]benzamide (Compound A37) was synthesized.

[0477]

[0478] Synthesis of Compound A37-1

[0479] A37-SM (10 g, 35.28 mmol) and 1,2-dibromoethanol (7.3 g, 38.81 mmol) were dissolved in DMF (100 mL), K2CO3 (10 g, 70.57 mmol) was added, and the reaction was carried out at 80 ° C. The reaction was stopped after 16 h. H2O was added to the reaction solution, and EA (50 mL x 3) was used for extraction. The organic phase was concentrated and purified by column chromatography (PE: EA = 30: 1). Compound A37-1 (9.7 g, white solid, purity 90.31%) was obtained. Yield: 63.6%. LCMS (ESI) m / z calcd. for C 17 H 32 BrN3O2[M+H] + 390.2; found 395.2. It can be seen that the product structure is correct.

[0480] Synthesis of Compound A37-2

[0481] A37-1 (9.7 g, 24.85 mmol) and 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (7.1 g, 27.33 mmol) were dissolved in DMF (100 mL), K2CO3 (6.87 g, 49.7 mmol) was added and stirred for 5 min, and then reacted at 80°C. The reaction was stopped after 16 h. H2O was added to the reaction solution, and extracted with EA (50 mL x 3). The organic phase was concentrated and purified and separated by column chromatography (DCM: MeOH = 20: 1). Compound A37-2 (11.4 g, yellow solid, purity 87.31%) was obtained. Yield: 70.4%. LCMS (ESI) m / z calcd.for C 30 H 44 N6O5[M+H] + 569.3; found 569.4. It can be seen that the product structure is correct.

[0482] Synthesis of compound A37-3

[0483] Under magnetic stirring, in a 100mL single-mouth bottle, A37-2 (10g, 17.58mmol) was dissolved in DCM (100mL). The solution was light yellow and turbid. HCl in 1,4-dioxane (20mL, 4M) was slowly added dropwise under ice bath conditions. The reaction was stopped after 4h. After the reaction was completed, the mixture was filtered and dried. Compound A37-3 (8.1g, light yellow solid, purity 88.21%) was obtained. Yield: 91.2%.

[0484] Synthesis of Compound A37-4

[0485] A2-SM (500 mg, 1.81 mmol) was dissolved in DMF (8 mL), DIEA (700 mg, 5.43 mmol) and HATU (690 mg, 1.81 mmol) were added, and after stirring for 5 min, 3-(trifluoromethyl)aniline (435 mg, 2.72 mmol) dissolved in DMF (2 mL) was added, and the reaction was stopped at room temperature for 16 h. H2O was added to the reaction solution, and extracted with EA (30 mL x 3). The organic phase was concentrated and purified and separated by column chromatography (PE:EA=10:1) to obtain compound F6-1 (520 mg, white solid, purity 90.33%). Yield: 61.8%. LCMS (ESI) m / z calcd.for C 16 H 13 F3INO[M+H] + 420.0; found 420.1. It can be seen that the product structure is correct.

[0486] Synthesis of Compound A37-5

[0487] Compound A37-4 (500 mg, 1.19 mmol), A34-12 (290 mg, 1.79 mmol), CuI (50 mg, 0.238 mmol), Pd(PPh3)2Cl2 (13 mg, 0.119 mol), DIEA (470 mg, 3.57 mmol) and DMF (5 mL) were added to a 100 mL single-mouth bottle in sequence. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (30 mL) and extracted three times with EA (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A37-5 (400 mg, brown oil, purity 92.13%) was obtained. Yield: 68.29%. LCMS (ESI) m / z calcd.for C 24 H 16 F4N4O[M+H] +453.1; found 453.2. It can be seen that the product structure is correct.

[0488] Synthesis of compound A37

[0489] A37-5 (400 mg, 0.885 mmol) and A37-3 (536 mg, 1.06 mmol) were dissolved in DMF (5 mL), K2CO3 (245 mg, 1.77 mmol) was added, and the reaction was carried out at 80°C. The reaction was stopped after 16 h. H2O was added to the reaction solution, and the mixture was extracted with DCM (20 mL x 3). The organic phase was concentrated and purified by column chromatography (DCM: MeOH = 5: 1) to obtain compound A37 (72 mg, white solid, purity 93.11%). Yield: 8.42%. LCMS (ESI) m / z calcd. for C 49 H 51 F3N 10 O4[M+H] + 901.4; found 901.5. It can be seen that the product structure is correct.

[0490] Embodiment 38

[0491] In this example, compound 3-({2-[(2-{4-[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yl}ethyl)oxy]pyrazolo[1,5-a]pyrimidin-6-yl}ethynyl)-4-ethyl-N-[3-(imidazol-1-yl)phenyl]benzamide (Compound A38) was synthesized.

[0492]

[0493] Synthesis of Compound A38-1

[0494] Hydroxyacetaldehyde dimethyl acetal (10 g, 94.23 mmol) was dissolved in DMF (50 mL), and NaH (2.7 g, 113.1 mmol) was added. After stirring for 30 min, A34-12 (15.2 g, 94.23 mmol) was added. The reaction was stopped after reacting at room temperature for 4 h. H2O was added to the reaction solution, and EA (50 mL x 3) was used for extraction. The organic phase was concentrated and purified by column chromatography (PE: EA = 10: 1) to obtain compound F7-11 (13 g, off-white solid, purity 91.31%). Yield: 50.9%. LCMS (ESI) m / z calcd.forC 12 H 13 N3O3[M+H] + 248.1; found 248.2. It can be seen that the product structure is correct.

[0495] Synthesis of Compound A38-2

[0496] A2-SM (10 g, 36.22 mmol) was dissolved in DMF (40 mL), TEA (11 g, 108.7 mmol) and HATU (13 g, 36.22 mmol) were added, and after stirring for 5 min, 3-(imidazol-1-yl)aniline (6.4 g, 39.8 mmol) dissolved in DMF (1 mL) was added, and the reaction was stopped after reacting at room temperature for 20 h. H2O was added to the reaction solution, and extracted with EA (50 mL x 3), and the organic phase was concentrated and purified and separated by column chromatography (PE: EA = 10: 1). Compound A38-2 (7.9 g, white solid, purity 86.31%) was obtained. Yield: 45.1%. LCMS (ESI) m / z calcd.for C 18 H 16 IN3O[M+H] + 418.0 found 418.1. It can be seen that the product structure is correct.

[0497] Synthesis of compound A38-3

[0498] Compound A38-2 (2 g, 4.79 mmol), A38-1 (1.78 g, 7.19 mmol), CuI (185 mg, 0.958 mmol), Pd (PPh3) 2Cl2 (170 mg, 0.240 mol), DIEA (1.9 g, 14.37 mmol) and DMF (30 mL) were added to a 250 mL single-mouth bottle in sequence. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a saturated sodium bicarbonate aqueous solution (50 mL) and extracted three times with EA (50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Compound A38-3 (1.8 g, brown oil, purity 88.21%) was obtained. Yield: 61.73%. LCMS (ESI) m / z calcd.for C 30 H 28 N6O4[M+H] + 537.2; found 537.3. It can be seen that the product structure is correct.

[0499] Synthesis of Compound A38-4

[0500] At 0°C, in a 100mL single-mouth bottle, A38-3 (1.8g, 3.35mmol) was dissolved in DCM (40mL), and the solution was light yellow and turbid. TFA (1mL) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred for 20min and then moved to room temperature for reaction. The reaction was stopped after 1h. After the reaction was completed, H2O was added to the reaction solution, and it was extracted with EA (30mLx3). The organic phase was concentrated to obtain compound A38-4 (1.2g, light yellow solid, purity 81.67%). Yield: 88.52%.

[0501] Synthesis of Compound A38

[0502] A38-4 (200 mg, 0.408 mmol) and A32-12 (150 mg, 0.408 mmol) were dissolved in THF (10 mL), AcOH (30 mg, 0.408 mmol) was added, and NaB(OAc)3 (200 mg, 3.06 mmol) was added after stirring for 5 min. The reaction was stopped at room temperature for 16 h. Dichloromethane and saturated sodium carbonate were added for extraction, and the organic layer was washed with saturated brine, dried, and spin-dried. It was purified and separated by column chromatography (DCM: MeOH = 10: 1). Compound A38 (28 mg, white solid, purity 96.36%) was obtained. Yield: 8.24%. LCMS (ESI) m / z calcd.for C 45 H 42 N 10 O5[M+H] + 803.3; found 803.4. It can be seen that the product structure is correct.

[0503] Embodiment 39

[0504] In this example, compound 5-({[3-({2-[(1-{1-[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]azetidin-3-yl}pyridin-4-yl)amino]pyrimidin-5-yl}ethynyl)-4-methoxyphenyl]carbonyl}amino)pyridine-3-carboxamide (Compound A39) was synthesized.

[0505]

[0506] Synthesis of Compound A39-1

[0507] A39-SM (2 g, 14.5 mmol) was added to a 100 mL single-mouth bottle, and thionyl chloride (10 mL) and DMF (2 drops) were added. The reaction solution was stirred at 65 ° C for 2 h. After the reaction was completed, the reaction solution was spin-dried and DCM was dragged 3 times to obtain a light yellow solid. At 0 ° C, THF (15 mL) was added to the above residue and slowly added dropwise to a THF (30 mL) solution of ammonia water (6 g, 43.5 mmol). After the dropwise addition, the reaction solution was warmed to room temperature and continued to stir for 2 hours. After the reaction was completed, the reaction solution was filtered and dried with water. A39-1 (1.4 g, light yellow solid, purity 88.7%) was obtained, with a yield of 62.5%. LCMS (ESI) m / z calcd. For C6H7N3O [M + H] + 138.1; found 138.2. It can be seen that the product structure is correct.

[0508] Synthesis of Compound A39-2

[0509] A15-5 (1.5 g, 3.31 mmol) was added to a 100 mL single-mouth bottle, followed by DMF (35 mL), followed by DIEA (500 mg, 4.42 mmol), HATU (680 mg, 3.31 mmol), stirred for 10 min, and A39-1 (500 mg, 3.65 mmol) was added. The temperature was raised to 40 °C and the reaction was stopped after 18 h. The reaction solution was poured into 30 mL of water, filtered to obtain a filter cake, dried, and 10 mL of methanol was added to slurry and filter to obtain a filter cake. Compound A39-2 (1 g, off-white solid, purity: 90.1%) was obtained by drying. Yield: 52.3%. LCMS (ESI) m / z calcd.ForC 30 H 33 N7O5[M+H] + 572.3; found 572.3. It can be seen that the product structure is correct.

[0510] Synthesis of compound A39-3

[0511] A39-2 (1 g, 1.75 mmol) and dichloromethane (20 mL) were added to a 50 mL single-mouth bottle, and HCl in 1,4-dioxane (4 M, 4 mL) was added dropwise. The reaction was stopped after 4 h. A large amount of solid precipitated, which was filtered (rinsed with DCM), and the filter cake was dried to obtain compound A39-3 (740 mg, yellow solid), with a yield of 89.7%.

[0512] Synthesis of Compound A39

[0513] A39-3 (150 mg, 0.318 mmol) and A40-1 (150 mg, 0.350 mmol) were dissolved in THF (10 mL), AcOH (22 mg, 0.318 mmol) was added, and NaB (OAc) 3 (200 mg, 3.18 mmol) was added after stirring for 5 min. The reaction was reacted at room temperature, and the reaction progress was monitored by TLC (developing solvent: DCM: MeOH = 5: 1). The reaction was stopped after 16 h. Dichloromethane and saturated sodium bicarbonate were added for extraction, and the organic layer was washed with saturated brine, dried, and spin-dried. The prepared plate was purified and separated (DCM: MeOH = 5: 1). Compound A39 (38 mg, white solid, purity 95.16%) was obtained. Yield: 14.7%. LCMS (ESI) m / z calcd.for C 41 H 40 N 10 O6[M+H] + 769.3; found 769.3. It can be seen that the product structure is correct.

[0514] Embodiment 40

[0515] In this example, compound 3-({[3-({2-[(1-{1-[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]azetidin-3-yl}pyridin-4-yl)amino]pyrimidin-5-yl}ethynyl)-4-methoxyphenyl]carbonyl}amino)-5-(trifluoromethyl)benzene-1-carboxamide (Compound A40) was synthesized.

[0516]

[0517] Synthesis of Compound A40-1

[0518] At room temperature, A33-SM2 (1 g, 3.09 mmol) and azetidin-3-one hydrochloride (1 g, 3.71 mmol) were dissolved in 1,4-dioxane (50 mL), and C S CO3 (3 g, 9.27 mmol), and then Pd-PEPPSI-IHeptCl (280 mg, 0.309 mmol) was added. The argon gas was replaced 3 times, and the temperature was raised to 100°C for 18 h to stop the reaction. Saturated NaCl aqueous solution was added, and DCM (50 mL×3) was used for extraction. The organic phases were combined, dried and concentrated, and purified by column chromatography (DCM: MeOH = 100: 1 / 20: 1) to obtain compound A40-1 (400 mg, light yellow solid, purity 89.21%). Yield: 36.8%. LCMS (ESI) m / z calcd.ForC 16 H 15N3O4[M+H] + 314.1; found 314.2. It can be seen that the product structure is correct.

[0519] Synthesis of Compound A40

[0520] A40-1 (128 mg, 0.408 mmol) and A24-2 (200 mg, 0.372 mmol) were dissolved in THF (10 mL), AcOH (28 mg, 0.372 mmol) was added, and NaB (OAc) 3 (80 mg, 0.372 mmol) was added after stirring for 5 min. The reaction was allowed to react at room temperature and the reaction was stopped after 16 h. Dichloromethane and saturated sodium bicarbonate were added for extraction, and the organic layer was washed with saturated brine, dried, and spin-dried. The organic layer was purified and separated by a preparative plate (DCM: MeOH = 10: 1). Compound A40 (42 mg, white solid, purity 95.77%) was obtained. Yield: 12.96%. LCMS (ESI) m / z calcd. For C 43 H 40 F3N9O6[M+H] + 836.3; found 836.4. It can be seen that the product structure is correct.

[0521] Embodiment 41

[0522] In this example, the compound N-[5-chloro-3-(trifluoromethyl)phenyl]-3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxybenzamide (Compound A41) was synthesized.

[0523]

[0524] Synthesis of Compound A41-1

[0525] A16-3 (1.5 g, 3.32 mmol) was added to a 25 mL single-mouth bottle, followed by DMF (15 mL), TEA (1 g, 3.32 mmol), HATU (1.3 g, 3.32 mmol), 3-chloro-5-(trifluoromethyl)aniline (760 mg, 3.98 mmol), the temperature was raised to 30 ° C, and the reaction was stopped after 18 h. The reaction solution was poured into 30 mL of water, filtered to obtain a filter cake, dried, and 10 mL of methanol was added to slurry and filter to obtain a filter cake. Compound A41-1 (920 mg, off-white solid, purity 91.3%) was obtained by drying. Yield: 39.9%. LCMS (ESI) m / z calcd. For C 31 H31 ClF3N5O4[M+H] + 630.2; found 630.3. It can be seen that the product structure is correct.

[0526] Synthesis of Compound A41-2

[0527] A41-1 (900 mg, 0.94 mmol) and dichloromethane (20 mL) were added to a 50 mL single-mouth bottle, and HCl in 1,4-dioxane (4 M, 4 mL) was added dropwise. The reaction was stopped after 4 h. A large amount of solid was precipitated, which was filtered (rinsed with DCM), and the filter cake was dried to obtain compound A41-2 (710 mg, yellow solid), with a yield of 93.7%.

[0528] Synthesis of Compound A41

[0529] A41-2 (200 mg, 0.377 mmol) and A15-SM2 (223 mg, 0.566 mmol) were added to a 50 mL single-mouth bottle, and acetonitrile (15 mL) was added to partially dissolve. After adding potassium carbonate (250 mg, 1.57 mmol), the temperature was raised to 60 ° C and the reaction was stopped after 24 hours. The reaction liquid was filtered and the filtrate was concentrated. After column chromatography, the eluent ratio was DCM: MeOH = 100: 1 to 10: 1 to obtain 140 mg of yellow solid. Compound A41 (37 mg, off-white solid, 97.4%) was obtained by TLC plate. Yield: 11.3%. LCMS (ESI) m / z calcd.ForC 42 H 38 ClF3N8O6[M+H] + 843.3; found 843.4. It can be seen that the product structure is correct.

[0530] Embodiment 42

[0531] In this example, compound 3-[({3-[(2-{[1-(3-{[2-(2,6-dioxypyridin-3-yl)-1-oxypyridin-2,3-dihydro-1H-isoindol-5-yl]amino}-3-oxypyridin-4-yl]amino}pyrimidin-5-yl)ethynyl]-4-methoxyphenyl}carbonyl)amino]-N,N-dimethyl-5-(trifluoromethyl)benzamide (Compound A42) was synthesized.

[0532]

[0533] Synthesis of Compound A42-1

[0534] A16-3 (1.5 g, 3.32 mmol) was added to a 25 mL single-mouth bottle, DMF (15 mL) was added, TEA (1 g, 3.32 mmol) was added dropwise, HATU (1.3 g, 3.32 mmol) was added in batches, and stirred for 10 min. 3-amino-N, N-dimethyl-5-(trifluoromethyl)benzamide (812 mg, 3.98 mmol) was added, and the reaction was stopped after 18 h at 30 ° C. The reaction solution was poured into 30 mL of water, filtered to obtain a filter cake, dried, and 10 mL of methanol was added to slurry and filter to obtain a filter cake. Compound A42-1 (1.2 g, off-white solid) was dried. Yield: 55.0%. LCMS (ESI) m / z calcd.For C 34 H 37 F3N6O5[M+H] + 667.3; found 667.4. It can be seen that the product structure is correct.

[0535] Synthesis of Compound A42-2

[0536] A42-1 (600 mg, 0.94 mmol) and dichloromethane (20 mL) were added to a 50 mL single-mouth bottle, and HCl in 1,4-dioxane (4 M, 4 mL) was added dropwise. The reaction was stopped after reacting at room temperature for 4 h. A large amount of solid precipitated, which was filtered (rinsed with DCM), and the filter cake was dried to obtain compound A42-2 (550 mg, yellow solid), with a yield of 94.0%.

[0537] Synthesis of Compound A42

[0538] A42-2 (300 mg, 0.52 mmol) and A15-SM2 (270 mg,

[0539] 0.68mmol), acetonitrile (15mL), potassium carbonate (250mg, 1.57mmol) were added, the temperature was raised to 60℃ for 24h, and the reaction was stopped. The reaction solution was filtered, and the filtrate was concentrated and subjected to column chromatography. The eluent ratio was DCM:MeOH=100:1-10:1 to obtain compound A42 (58mg, off-white solid, 92.7%), with a yield of 11.2%. LCMS (ESI) m / z calcd.ForC 45 H 44 F3N9O7[M+H] + 880.3; found 880.2. It can be seen that the product structure is correct.

[0540] Embodiment 43

[0541] In this example, the compound N-(3-carbamoyl-5-(trifluoromethyl)phenyl)-3-[(2-(((1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)-2-oxoethyl)piperidin-4-yl)amino)pyrimidin-5-yl)ethynyl]-4-methoxybenzamide (Compound A43) was synthesized.

[0542]

[0543] Synthesis of Compound A43-1

[0544] A43-SM (1 g, 3.86 mmol) was added to a 100 mL three-necked flask, dissolved in THF (25 mL), replaced with argon three times, and bromoacetyl chloride (2 g, 11.57 mmol) was added dropwise at 0°C. The reaction solution was reacted at 60°C for 12 h. After the reaction was completed, methanol (1 mL) was added dropwise under an ice-water bath, stirred for 5 min, and concentrated. The residue was dissolved in methanol, EA was added dropwise to precipitate the solid, and the compound A43-1 (800 mg, off-white solid) was obtained by filtration. The yield was 50.6%. LCMS (ESI) m / z calcd.for C 15 H 14 BrN3O4[M+H]+380.0; found380.1. It can be seen that the product structure is correct.

[0545] Synthesis of Compound A43

[0546] A43-1 (300 mg, 0.52 mmol) and A24-2 (250 mg, 0.68 mmol) were added to a 50 mL single-mouth bottle, and acetonitrile (15 mL) was added to partially dissolve. After adding potassium carbonate (250 mg, 1.57 mmol), the temperature was raised to 60 ° C for reaction. TLC monitored the progress of the reaction (developing solvent: DCM: MeOH = 10: 1), and the reaction was stopped after 24 hours. The reaction solution was filtered, and the filtrate was concentrated and subjected to column chromatography with an eluent ratio of DCM: MeOH = 100: 1 to 10: 1 to obtain 200 mg of a yellow solid. Compound A43 (77 mg, off-white solid, 95.7%) was obtained by preparation plate. Yield: 18.2%. LCMS (ESI) m / z calcd. For C 42 H 38 F3N9O7[M+H] + 838.3; found 838.2. 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),10.62(s,1H),10.11(s,1H),8.55(s,1H),8.45(d,J=21.8Hz,3H),8.31–8.16( m,2H),8.09–7.99(m,2H),7.96(s,1H),7.76–7.66(m,3H),7.61(s,1H),7.27(d,J=8.9Hz,1H),5.09(dd,J=13.3,5.1H z, 1H), 4.44 (d, J = 17.4 Hz, 1H), 4.30 (d, J = 17.4 Hz, 1H), 3.95 (s, 3H), 3.77 (s, 1H), 3.22–3.16 (m, 2H), 2.95–2.85 (m, 3H), 2.60 (d, J = 17.2 Hz, 1H), 2.28 (t, J = 11.3 Hz, 3H), 2.05–1.96 (m, 1H), 1.89 (d, J = 19.1 Hz, 2H), 1.66 (t, J = 11.2 Hz, 2H). It can be seen that the product structure is correct.

[0547] The HR-MS calculated values ​​and HR-MS measured values ​​of compounds A1-A43 synthesized in the above Examples 1-43 of the present invention are summarized in the following Table 1, respectively.

[0548] Table 1 HR-MS values ​​of representative compounds

[0549]

[0550]

[0551] Experimental example

[0552] The following experimental examples of the present invention are used to measure the biological activity and study the mechanism of representative compounds.

[0553] 1. Degradation of target protein DDR1 and inhibition of DDR1 phosphorylation

[0554] This example analyzes the degradation effect of each compound on the target protein DDR1 and the inhibitory effect on DDR1 phosphorylation.

[0555] This example measures the activity of the representative compounds synthesized in the above examples in degrading DDR1 in the tumor cell line T47D. The activity is shown as the half degradation concentration (DC50-DDR1); when the half degradation concentration is less than or equal to 100nM, the compound activity is marked as "++++"; when the half degradation concentration is greater than 100nM and less than 500nM, the compound activity is marked as "+++"; when the half degradation concentration is greater than 500nM and less than 1000nM, the compound activity is marked as "++"; when the half degradation concentration is greater than 1000nM and less than 3000nM, the compound activity is marked as "+"; when the half degradation concentration is greater than 3000nM, the compound activity is marked as "-".

[0556] This example measures the activity of the representative compounds synthesized in the above examples in inhibiting DDR1 phosphorylation in the tumor cell line T47D. The activity is shown as half inhibitory concentration (IC50-pDDR1); when the half degradation concentration is less than or equal to 100nM, the compound activity is marked as "++++"; when the half degradation concentration is greater than 100nM and less than 500nM, the compound activity is marked as "+++"; when the half degradation concentration is greater than 500nM and less than 1000nM, the compound activity is marked as "++"; when the half degradation concentration is greater than 1000nM and less than 3000nM, the compound activity is marked as "+"; when the half degradation concentration is greater than 3000nM, the compound activity is marked as "-".

[0557] Western blot experimental process: breast cancer cells (T47D) were inoculated into 6-well plates at a concentration of 2*105 cells / mL, 2 mL was added to each well, and incubated for 24 hours. The cells were observed under a microscope the next day to confirm the cell status. Compounds were prepared, and a certain amount of representative compounds or control drugs in the above embodiments of the present invention or cabozantinib were weighed respectively, and a mother solution with a higher concentration was first prepared with pure DMSO. The compound concentrations of the solution containing 0.5% DMSO were prepared by gradient dilution: 2mM, 200μM, 100μM, 20μM, 2μM, 200nM, 0nM solution, and the solution was shaken to mix thoroughly. Take a 6-well plate, discard the supernatant, and add PBS to wash the cells 3 times. Add 2ml of type I collagen with a final concentration of 50ug / ml, and add 10μl of solution to each well. After incubation for 24 hours, remove the cells and wash the cells 3 times with pre-cooled PBS. Carefully remove the culture medium and leave the cell pellet, add 2000μl RIPA lysis buffer premixed with 1% PMSF and 1% phosphatase inhibitor, shake well to make the cells fully contact, and incubate on ice for 30min. Scrape the lysate with a cell scraper and transfer it to a 1.5ml centrifuge tube. Centrifuge at 12000rpm for 10min, aspirate the supernatant, and use the BCA protein quantification kit to determine and normalize the sample loading amount.

[0558] Mix BCA-A and B solutions in a ratio of 50:1 to prepare the working solution. Dilute the sample 5 times and then pipette 20ul into a 96-well ELISA plate. Add 200μl of premixed BCA working solution and incubate at 37℃ for 20min. Measure the absorbance at A562nm with an ELISA reader and substitute it into the protein standard curve formula to obtain the sample concentration. After calculating the volume required for loading 50μg, dispense the sample into a 1.5ml centrifuge tube. Add 10μl of loading buffer to each tube and mix well. Heat in a 100℃ water bath for 5min to denature the protein.

[0559] Take a 4-20% gradient precast gel and fix it on the gel rack, add electrophoresis solution to the tank, remove the comb, load the marker and protein sample, and adjust the voltage to 120V after 80V electrophoresis for 30 minutes. Stop electrophoresis when the marker band reaches the forefront of the gel. Take out the gel block and cut it to a suitable size. Cut the PVDF membrane and activate it in methanol. Place the membrane and gel in the filter paper in the filter screen on the splint. Place the splint in the electrotransfer tank and electrotransfer at 250mA constant current for 2.5h. Remove the membrane and place it in 5% skim milk powder dissolved in 1×TBST for 2h. Cut the membrane according to the molecular weight of the target protein and the internal reference protein, place it in the primary antibody diluted in 1xTBST, and shake it overnight at 4°C. The next day, the strips were transferred to 1xTBST and washed three times, and then the secondary antibody diluted in a specific ratio was added and incubated with shaking for 1.5 h. The strips were transferred to 1×TBST again and washed three times, and then ECL color developing solution was prepared. The membrane was placed in an imaging system, the surface was covered with ECL color developing solution, and the membrane was photographed using the imaging system.

[0560] ImageJ was used to analyze the results, and the gray values ​​of each band of the target protein and the internal reference protein were measured. After the results were normalized by the target protein gray value / GAPDH gray value of the corresponding lane, quantitative analysis was performed according to the protein amount = gray value of the experimental group / gray value of the blank control group.

[0561] The performance results of the representative compounds synthesized in the above examples are shown in Table 2 below, and the degradation and phosphorylation inhibition effects of the representative compounds on DDR1 are shown in the attached table. Figure 1 .

[0562] Table 2 Inhibition of DDR1 phosphorylation and degradation by representative compounds

[0563]

[0564]

[0565] The experimental results confirm that the compounds defined in the present invention show the activity of degrading and / or inhibiting the target protein DDR1 in tumor cells.

[0566] 2. Inhibitory effect on cancer cell growth

[0567] This example measures the growth inhibitory effect of the above representative compounds on cancer cells.

[0568] Take human chronic myeloid leukemia cells (K562) and inoculate them into 96-well plates at a concentration of 4×104 cells / mL, add 100μL to each well, and incubate for 30 minutes. Observe the cell state under a microscope. Prepare the compounds, weigh a certain amount of the series of compounds of this patent, first prepare a mother solution with a higher concentration using pure DMSO, and use a gradient dilution method to prepare concentrations of 200μM, 60μM, 20μM, 6μM, 2μM, 600nM, 200nM, and 0nM solutions, and shake to mix them thoroughly.

[0569] After preparation, add 1 μL of drug solution to each well of a 96-well plate, and then add 100 μL of culture medium, and repeat three times for each dose. Gently shake to allow the cells and drug solution to fully contact, and incubate in an incubator for 72 hours. Take out after 72 hours, use the CCK-8 kit to develop color, add 20 μL of CCK-8 reagent to each well, shake gently, and incubate in the incubator for one hour. Take out after one hour, use an ELISA reader to measure the absorbance of each well at a wavelength of 450 nm, and calculate according to the following formula to obtain the growth inhibition ability of the compound on gastric cancer cells at the current dose and time:

[0570] Cell growth inhibition rate = [1 - (absorbance of experimental group - absorbance of culture medium control group) / (absorbance of blank control group - absorbance of culture medium control group)] × 100%.

[0571] The half maximum inhibitory concentration (IC50) was calculated using GraphPad Prism 9 software (GraphPad Software, La Jolla, CA). In the following results, when the half inhibition concentration is less than or equal to 500 nM, the compound activity is marked as "++++"; when the half inhibition concentration is greater than 500 nM and less than 1000 nM, the compound activity is marked as "+++"; when the half inhibition concentration is greater than 1000 nM and less than 3000 nM, the compound activity is marked as "++"; when the half inhibition concentration is greater than 3000 nM and less than 10000 nM, the compound activity is marked as "+"; when the half inhibition concentration is greater than 10000 nM, the compound activity is marked as "-".

[0572] The results of representative compounds measured in this example are shown in Table 3 below.

[0573] Table 3 Results of the inhibitory effects of compounds on cancer cell growth

[0574]

[0575]

[0576] It can be seen that the compounds defined in the present invention have a significant effect on tumor cell proliferation, with an IC50 value of less than 2 μM.

[0577] 3. Study on degradation mechanism

[0578] This example conducts research experiments on the mechanism by which the above representative compounds degrade DDR1 through the ubiquitin proteasome pathway.

[0579] Breast cancer cells (T47D) were inoculated into 6-well plates at a concentration of 2×105 cells / mL, 2 mL was added to each well, and incubated for 24 hours. The cells were observed under a microscope the next day to confirm the cell status. The compound of this patent and the proteasome inhibitor (MG132) and ubiquitination inhibitor (MLN4924) were prepared. Take a 6-well plate, add MG132 and MLN4924 with a final concentration of 5mM to the wells according to the experimental settings, and then add the compound of this patent. After incubation for 24 hours, take out the cells and wash the cells 3 times with pre-cooled PBS. Carefully aspirate the culture medium to leave the cell pellet, add 200μl RIPA lysis buffer premixed with 1% PMSF and 1% phosphatase inhibitor, shake well to make the cells fully contact, and incubate on ice for 30 minutes. Use a cell scraper to scrape the lysate and transfer it to a 1.5ml centrifuge tube, centrifuge at 12000rpm for 10 minutes, aspirate the supernatant, and use the BCA protein quantification kit to determine and homogenize the sample loading.

[0580] Mix BCA-A and B solutions in a ratio of 50:1 to prepare the working solution. Dilute the sample 5 times and then pipette 20ul into a 96-well ELISA plate. Add 200μl of premixed BCA working solution and incubate at 37℃ for 20min. Measure the absorbance at A562nm with an ELISA reader and substitute it into the protein standard curve formula to obtain the sample concentration. After calculating the volume required for loading 50μg, dispense the sample into a 1.5ml centrifuge tube. Add 10μl of loading buffer to each tube and mix well. Heat in a 100℃ water bath for 5min to denature the protein.

[0581] Take a 4-20% gradient precast gel and fix it on the gel rack, add electrophoresis solution to the tank, remove the comb, load the marker and protein sample, and adjust the voltage to 120V after 80V electrophoresis for 30 minutes. Stop electrophoresis when the marker band reaches the forefront of the gel. Take out the gel block and cut it to a suitable size. Cut the PVDF membrane and place it in methanol for activation. Place the membrane and gel in the filter paper in the filter screen on the splint. Place the splint in the electrotransfer tank and electrotransfer at 250mA constant current for 2.5h. Remove the membrane and place it in 5% skim milk powder dissolved in 1xTBST for 1h. Cut the membrane according to the molecular weight of the target protein and the internal reference protein, place it in the primary antibody diluted in 1xTBST, and shake it overnight at 4°C. The next day, the strips were transferred to 1xTBST and washed three times, and then the secondary antibody diluted in a specific ratio was added and incubated with shaking for 1.5h. The strips were transferred to 1xTBST again and washed three times, and then the ECL color developing solution was prepared. The membrane was placed in an imaging system, the surface was covered with the ECL color developing solution, and the membrane was photographed using the imaging system.

[0582] The results of the representative compounds verified in this example are shown in the attached Figure 2 shown.

[0583] It can be seen that the compounds defined in the present invention can degrade DDR1, and this effect can be antagonized by a proteasome inhibitor (MG132) or a ubiquitination inhibitor (MLN4924). It also confirms that the target protein degradation effect of the compounds defined in the present invention is through the ubiquitination proteasome system.

[0584] 4. Inhibition rate of kinase

[0585] In this example, the inhibition rate of the representative compounds synthesized in the above examples on 76 kinases was measured.

[0586] In this example, the representative compound A15 synthesized above was used to conduct affinity inhibition experiments on 76 kinases using HTRF Kinase Assay and ADP-Glo ​​Kinase Assay.

[0587] The HTRF KinaseAssay affinity inhibition experiment includes the following steps:

[0588] (1) Prepare 2× ATP+substrate solution and 2× kinase / metal ion solution using assay buffer;

[0589] (2) 50 nL of compound was transferred to the 384 assay plate using Echo 655. After centrifugation, 2.5 μL of 2× kinase solution was added to the 384 assay plate and incubated at 25°C for 10 min.

[0590] (3) Add 2.5 μL of 2× substrate + ATP solution to the wells and incubate for 60 min;

[0591] (4) Prepare 2×XL665 and antibody solution with detection buffer;

[0592] (5) Add 5 μL of kinase detection reagent and incubate at 25°C for 60 min;

[0593] (6) Use a BMG microplate reader to read the fluorescence signals at 620 nm (Cryptate) and 665 nm (XL665).

[0594] The ADP-Glo ​​KinaseAssay affinity inhibition experiment includes the following steps:

[0595] (1) Prepare 2× ATP + substrate solution and 2× kinase / metal solution using assay buffer;

[0596] (2) 40 nL of compound was transferred to the 384 assay plate using Echo 655, 2 μL of 2× kinase solution was added and mixed, and incubated at 25°C for 10 min in a polystyrene-coated 384 assay plate;

[0597] (3) Add 2 μL of 2× substrate + ATP solution to the wells and incubate for 60 min;

[0598] (4) Add 4 μL of ADP-Glo ​​reagent to the wells and incubate at 25°C for 40 min;

[0599] (5) Add 8 μL of kinase detection reagent and incubate at 25°C for 40 min;

[0600] (6) The fluorescence signal is recorded on an ELISA reader.

[0601] Data analysis: The readout value of the reaction control (1% DMSO) was set as 0% inhibition, and the readout value of the background (10 μM Positive control) was set as 100% inhibition. Then the inhibition percentage of each test solution was calculated. The calculation formula of %Inhibition is as follows:

[0602]

[0603] Wherein, Data positive: average ratio of positive control (10 μM), Data vehicle: average signal value of control (1% DMSO).

[0604] The dendrogram of the inhibition rate of the compounds in this example on 76 tyrosine kinases is shown in the attached figure. Figure 3 shown.

[0605] The experimental results show that the compounds defined in the present invention exhibit high selectivity for DDR1 at a concentration of 30 nM. 5. Comparison of activity and solubility of the compounds

[0606] This experimental example systematically studies the improvement of activity and physicochemical properties by structural modification of the representative compounds synthesized above. Compounds A4, A7, A10, and A11 were treated with methanolic hydrochloric acid solution to obtain their corresponding hydrochlorides. Among them, the demethyl derivatives A4-DeMe, A7-DeMe and their hydrochlorides of compounds A4 and A7 were prepared in a similar manner to A4 and A7; and the ethyl derivatives A10-Et, A11-Et and their hydrochlorides of A10 and A11 were prepared in a similar manner to A10 and A11.

[0607]

[0608] In this experimental example, the activity determination method refers to the method of the aforementioned experimental example, and the solubility determination is based on the method specified in the Chinese Pharmacopoeia Volume IV. When the solubility is dissolved, it is marked as "++++"; when the solubility is slightly soluble, it is marked as "+++"; when the solubility is slightly soluble, it is marked as "++"; when the solubility is very slightly soluble, it is marked as "+"; when the solubility is almost insoluble or insoluble, it is marked as "-".

[0609] The results of representative compounds measured in this example are shown in Table 4 below.

[0610] Table 4 Compound activity and solubility results

[0611] Compound Salt type <![CDATA[IC 50 -pDDR1]]> <h2 style=";text-align:left;direction:ltr"><![CDATA[IC <h2 style=";text-align:left;direction:ltr"> 50 <h2 style=";text-align:left;direction:ltr"> (K562)]]><h2 style=";text-align:left;direction:ltr"> Solubility A4 HCl ++++ ++ + A7 HCl ++++ +++ ++ A4-DeMe HCl ++ + - A7-DeMe HCl +++ + - A10 HCl ++++ ++++ +++ A11 HCl ++++ ++++ ++ A10-Et HCl +++ +++ + A11-Et HCl +++ ++ -

[0612] The above experimental results show that the introduction of a methyl group at the 3-position of the parent core structure pyrazolo[1,5-a]pyrimidine can significantly improve the activity of the compound. The introduction of a methoxy structure on the benzene ring of the benzamide structure not only significantly improves its activity, but also significantly improves its solubility.

[0613] In summary, the benzamide compounds of the present invention can effectively inhibit or degrade DDR1 compounds, and can regulate downstream key proteins by degrading or inhibiting discoidin domain receptor 1 (DDR1) and further play a role in treating related diseases.

[0614] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof, characterized in that: It has the structure shown in the following formula (I): in, The R a Selected from H, C1-C3 straight chain or branched alkyl, or C1-C3 straight chain or branched alkyl in which some or all of the hydrogen atoms are replaced by fluorine; The R b Selected from H or C1-C3 straight or branched alkyl; The G is selected from CR1 or N; The R c and R1 are independently selected from H, halogen, R2, -OR2, -(CH2) x NR3R4, -(CH2) y CONR3R4 or a 5-7 membered aromatic heterocycle containing at least one nitrogen atom; wherein R2 is selected from C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; wherein R3 or R4 are independently selected from H, C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; or, R3 and R4 and the nitrogen atom to which they are connected together form a 3-7 membered azacycloalkyl group containing 1-2 nitrogen atoms, and the hydrogen atoms on the nitrogen can be replaced by C1-C3 straight or branched alkyl; wherein x and y are independently selected from integers of 0-2; Ar is selected from aromatic rings wherein A and B are independently selected from N or CH, and at least one of them is N; wherein the asterisk (*) indicates the connection site with the M group, and the wavy line indicates the connection site with the alkynyl carbon; The M is selected from H, R5, -NHR5 or a functional group consisting of two parts EL-**, wherein the R5 is selected from C1-C3 straight chain or branched alkyl; the E is a ligand part capable of binding to the E3 binding enzyme cereblon, which is connected to the bridge part L through a covalent bond; wherein the double asterisk (**) indicates the asterisk connection site with the Ar group; The L portion is a bridging group connecting the E portion and the Ar portion.

2. The benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof according to claim 1, characterized in that: The E has the structure shown in E1 or E2 below: Wherein, D1 is selected from CH2 or C=O, preferably CH2; D2 is selected from CH or N; In the formula, the (#) position indicates the position where E1 or E2 is covalently bonded to the L portion, and the connection position is at position 1 or 2 of the E1 or E2 structural formula.

3. The benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof according to claim 2, characterized in that: The E is selected from at least one of the following structures:

4. The benzamide compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof, characterized in that: The L is selected from the structures L1-L7 shown below, wherein the wavy line indicates the position where L is covalently bonded to the asterisk position of the Ar part; the asterisk (*) indicates the position where L is covalently bonded to the pound position (#) of the E part; Wherein, m is selected from a natural number between 0 and 2, n is selected from a natural number between 0 and 1, and p is selected from a natural number between 1 and 2; Said T1 is selected from O, NH or CONH; Said T2 is selected from O or NH; Said T3 is selected from N or CH; The T4 is selected from NH or absent; The T5 is selected from O or CONH; The T6 is selected from O or NH; Said T7 is selected from CH2 or CO or does not exist; The T8 is selected from CO or does not exist; The Cy is selected from a 4-7 membered azetidinyl group, preferably an azetidinyl group containing one nitrogen atom, preferably azetidine or azetidinyl group.

5. The benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof according to claim 4, characterized in that: The L is selected from at least one of the following structures:

6. The benzamide compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof, characterized in that: The R c or R1 is independently selected from H, NH2, CF3, OCH3, N(CH3)2, CONH2, CH2N(CH3)2 or 7. The benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug according to any one of claims 1 to 6, characterized in that: The compound is selected from one of the following structures:

8. A benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof, characterized in that: It has the structure shown in the following formula (II): in, The R d is selected from H, C1-C3 straight chain or branched chain alkyl, or C1-C3 straight chain or branched chain alkyl in which some or all of the hydrogen atoms are replaced by fluorine; The R e Selected from H or C1-C3 straight or branched alkyl; The G' is selected from CR6 or N; The R f , R6 are independently selected from H, halogen, R7, -OR7, -(CH2) x NR8R9, -(CH2) y CONR8R9 or a 5-7 membered aromatic heterocycle containing at least one nitrogen atom; wherein R7 is selected from C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; wherein R8 or R9 are independently selected from H, C1-C3 straight or branched alkyl or C1-C3 straight or branched alkyl in which a part or all of the hydrogen atoms are replaced by fluorine; or, R8 and R9 and the nitrogen atom to which they are connected together form a 3-7 membered azacycloalkyl group containing 1-2 nitrogen atoms, and the hydrogen atoms on the nitrogen can be replaced by C1-C3 straight or branched alkyl; wherein x and y are independently selected from integers of 0-2; Ar' is selected from aromatic rings wherein A and B are independently selected from N or CH, and at least one of them is N; wherein the asterisk (*) indicates the connection site with the M group, and the wavy line indicates the connection site with the alkynyl carbon; The M' is selected from a C1-C3 straight or branched alkyl group or a functional group consisting of two parts, E'-L'-**, wherein the E' is a ligand part capable of binding to the E3 binding enzyme cereblon, which is connected to the bridge part L' via a covalent bond; wherein the double asterisk (**) indicates the asterisk connection site with the Ar' group; The L' portion is a bridge group connecting the E' portion and the Ar' portion.

9. The benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof according to claim 8, characterized in that: When Ar' is When M' is composed of two parts: EL-**; or, When Ar' is When M' is a C1-C3 straight chain or branched chain alkyl group or is composed of two parts, EL-**.

10. The benzamide compound or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof according to claim 8 or 9, characterized in that: The E' has the structure shown in the following E1' or E2': Wherein, D1 is selected from CH2 or C=O, preferably CH2; D2 is selected from CH or N; In the formula, the (#) position indicates the position where E1' or E2' is covalently bonded to the L' part, and the connection position is at position 1 or 2 of the E1' or E2' structural formula.

11. The benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof according to claim 10, characterized in that: The E' is selected from at least one of the following structures:

12. The benzamide compound according to any one of claims 8 to 11 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof, characterized in that: The L' is selected from the structure L1'-L7' shown below, wherein the wavy line indicates the position where L' is covalently bonded to the asterisk position of the Ar' part; the asterisk (*) indicates the position where L' is covalently bonded to the pound position (#) of the E part; Wherein, m is selected from a natural number between 0 and 2, n is selected from a natural number between 0 and 1, and p is selected from a natural number between 1 and 2; Said T1 is selected from O, NH or CONH; Said T2 is selected from O or NH; Said T3 is selected from N or CH; The T4 is selected from NH or absent; The T5 is selected from O or CONH; The T6 is selected from O or NH; Said T7 is selected from CH2 or CO or does not exist; The T8 is selected from CO or does not exist; The Cy is selected from a 4-7 membered azetidinyl group, preferably an azetidinyl group containing one nitrogen atom, preferably azetidine or azetidinyl group.

13. The benzamide compound or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof according to claim 12, characterized in that: The L' is selected from at least one of the following structures:

14. The benzamide compound according to any one of claims 8 to 13 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof, characterized in that: The R f or R6 are independently selected from H, NH2, CF3, OCH3, N(CH3)2, CONH2, CH2N(CH3)2 or 15. The benzamide compound according to any one of claims 8 to 14 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope-labeled substance or prodrug thereof, characterized in that: The compound is selected from one of the following structures:

16. A pharmaceutical composition, characterized in that Contains an active drug and a pharmaceutically acceptable carrier or excipient; The active drug comprises the benzamide compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope label or prodrug thereof, or the benzamide compound according to any one of claims 8 to 15 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, isotope label or prodrug thereof.

17. The pharmaceutical composition according to claim 16, characterized in that: The pharmaceutical composition further comprises at least one additional biologically active agent; Preferably, the bioactive agent comprises at least one of an anticancer agent, an immunomodulator, an immune checkpoint inhibitor, a kinase inhibitor, a corticosteroid or an anti-inflammatory agent.

18. Use of the compound according to any one of claims 1 to 7, the compound according to any one of claims 8 to 15, or the pharmaceutical composition according to claim 16 or 17 for preparing a pharmaceutical preparation for degrading and / or inhibiting discoidin domain receptor 1 (DDR1); Preferably, when the M or M' in the compound is a functional group consisting of two parts, E'-L'-**, the compound has the effect of degrading and inhibiting DDR1; Preferably, when in the compound, the M or M' is H or a C1-C3 straight chain or branched alkyl group, the compound has the effect of inhibiting DDR1.

19. The use according to claim 18, characterized in that The discoidin domain receptor 1 (DDR1) includes wild type and structural mutant types.

20. Use of the compound according to any one of claims 1 to 7, the compound according to any one of claims 8 to 14, or the pharmaceutical composition according to claim 16 or 17 for preparing a medicament for preventing, diagnosing or treating a disease related to discoidin domain receptor 1 (DDR1).

21. The use according to claim 20, characterized in that The discoidin domain receptor 1 (DDR1)-related diseases or disorders include cancers, immune-related diseases, fibrotic diseases, and neurodegenerative diseases associated with abnormal expression of DDR1.

22. The use according to claim 21, characterized in that The cancers include gastric cancer, intestinal cancer, esophageal cancer, head and neck cancer, lung cancer, liver cancer, brain cancer, breast cancer, colorectal cancer, skin cancer, thyroid cancer, prostate cancer, soft tissue cancer, endometrial cancer, uterine cancer, testicular cancer, cervical cancer, ovarian cancer, fallopian tube tumors, leukemia, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, renal cell carcinoma, bladder cancer, kidney cancer, pancreatic cancer, lymphoma, non-Hodgkin's lymphoma, melanoma, myeloproliferative disease, sarcoma, angiosarcoma, peripheral neuroepithelioma, glioma, astrocytoma, oligodendroglioma, ependymoma, glioblastoma, neuroblastoma, gangliocytoma, ganglioglioma, medulloblastoma, pineal cell tumor, meningioma, meningiosarcoma, neurofibroma or neurothecoma.

23. The use according to claim 21, characterized in that The immune-related diseases include rheumatoid arthritis, autoimmune encephalomyelitis, ankylosing spondylitis, axial spondyloarthritis, psoriasis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, recurrent oral ulcers, Kawasaki disease, spondyloarthritis, neuromyelitis optica, Behcet's disease, lupus nephritis, familial Mediterranean fever, ulcerative colitis, autoimmune hepatitis, asthma, and arteriosclerosis.

24. The use according to claim 21, characterized in that The fibrotic disease includes pulmonary fibrosis, liver fibrosis, renal fibrosis or Crohn's disease.

25. The use according to claim 21, characterized in that The neurodegenerative disease includes Alzheimer's disease, amyotrophic lateral sclerosis, ataxia telangiectasia, bovine spongiform encephalopathy, Creutzfeldt-Jakob disease, Huntington's disease, cerebellar atrophy, multiple sclerosis, Parkinson's disease, primary lateral sclerosis, spinal muscular atrophy, cerebral ischemia, spastic paraplegia or myasthenia gravis.

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