Azaindole derivative and application thereof

Abstract

The invention discloses an azaindole derivative compound as well as a preparation method and application thereof. The compound has CSF1R inhibitory activity and can be widely applied to drugs for treating cancers, tumors, autoimmune diseases, neurodegenerative diseases, metabolic diseases or metastatic diseases.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceuticals, and provides a class of azaindole-derived compounds, their preparation methods and pharmaceutical uses. Such compounds have CSF1R inhibitory activity and can be widely used in drugs for treating cancer, tumors, autoimmune diseases, neurodegenerative diseases, metabolic diseases or metastatic diseases. Background Art

[0002] CSF-1R is a tyrosine kinase transmembrane receptor and also belongs to the growth factor CSF-1 / platelet-derived growth factor (PDGF) receptor family. This receptor family has inherent tyrosine-specific protein kinase activity. CSF-1R is involved in the survival, proliferation, differentiation, recruitment and function of mononuclear phagocytes (such as microglia, macrophages, monocytes). IL-34 has biological properties similar to those of CSF-1, but their main differences come from different spatio-temporal regulations. In addition, CSF-1 can act through autocrine and paracrine manners, while IL-34 only acts locally. Small molecule inhibitors or antibodies developed targeting the CSF-1R target have the activity of inhibiting the survival and proliferation of microglia, inflammatory macrophages, and tumor-associated macrophages (TAMs), thereby exerting anti-tumor, immunomodulatory and neuroinflammation-reducing effects. Currently, they have been used in the drug development of various cancers, central neurodegenerative diseases and chronic graft-versus-host disease and other diseases.

[0003] High levels of CSF-1 have been reported in breast cancer, pancreatic cancer, prostate cancer, ovarian cancer, renal cancer and many other types of cancers. The overexpression of CSF-1 and its receptor CSF-1R in tumors is also associated with poor prognosis. Targeting the colony-stimulating factor 1 receptor (CSF-1R) / CSF-1 pathway can regulate the function of tumor-associated macrophages TAMs. Microglia mediate neuroinflammatory responses, and neuroinflammation plays a central role in the pathogenesis of various neurodegenerative diseases (MS, AD, ALS). Studies have shown that CSF1R inhibitors can inhibit and kill over-activated microglia in the nervous system with high affinity and selectivity, effectively block the neuroinflammatory response driving disease development, and slow down nerve damage to promote nerve cell repair. In addition to its important role in the field of tumors and central neurodegenerative diseases, CSF-1R inhibitors also have broad application prospects, covering multiple fields such as inflammation, metabolism and bone-related diseases. Therefore, the drug development targeting the CSF-1R target has become a research hotspot for scholars and pharmaceutical companies at home and abroad.

[0004] Compared with the early approved multi-target CSF-1R inhibitors such as Surufatinib, Pexidartinib, Vorolanib, etc., the new generation of CSF-1R inhibitors need to have better selectivity to reduce the side effects caused by off-target effects. At the same time, the pharmacokinetic properties of small molecule drugs also need to be further optimized. The purpose of this invention is to discover orally active CSF1R inhibitors with high activity, high selectivity, and excellent pharmacokinetic properties through the structural modification of Pexidartinib. Summary of the Invention

[0005] Technical problems to be solved: This invention provides a class of azaindole-derived compounds, which have CSF1R inhibitory activity and can be widely used in drugs for treating cancer, tumors, autoimmune diseases, neurodegenerative diseases, metabolic diseases or metastatic diseases.

[0006] Technical solution: A class of azaindole compounds represented by formula I, their stereoisomers or their pharmaceutically acceptable salts, hydrates or solvate compounds,

[0007]

[0008] wherein,

[0009] R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, difluoromethyl or C1-C6 alkyl;

[0010] R2 is selected from hydrogen, chlorine or fluorine;

[0011] R3 and R4 are each independently selected from hydrogen, fluorine, C1-C3 alkyl, or R3 and R4 together with the connected carbon atom form an alkyl ring;

[0012] A is selected from indole, substituted indole, pyridine, substituted pyridine, pyrrole, substituted pyrrole, pyrimidine or substituted pyrimidine;

[0013] n is selected from the numbers 0, 1, 2 or 3.

[0014] Preferably, wherein,

[0015] R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl or C1-C3 alkyl;

[0016] R2 is selected from hydrogen, chlorine or fluorine;

[0017] R3 and R4 are each independently selected from hydrogen, fluorine, C1-C3 alkyl, or R3, R4 and the connected carbon atom together form an alkyl three-membered ring or four-membered ring;

[0018] A is selected from indole, substituted indole, pyridine, substituted pyridine, pyrrole or substituted pyrrole;

[0019] n is selected from the numbers 0, 1 or 2.

[0020] Preferably, among them,

[0021] R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl or methyl;

[0022] R2 is selected from hydrogen or fluorine;

[0023] R3 and R4 are both hydrogen at the same time, or R3 and R4 are both fluorine at the same time, or R3, R4 and the connected carbon atom together form an alkyl three-membered ring;

[0024] A is selected from indole, pyridine, halogen-substituted pyridine, pyrrole or N-methylpyrrole;

[0025] n is selected from the numbers 0 or 1.

[0026] Preferably, the compound is:

[0027]

[0028] Compound 1: As shown in S1;

[0029]

[0030] Compound 2: As shown in S2;

[0031]

[0032] Compound 3: As shown in S3;

[0033]

[0034] Compound 4: As shown in S4;

[0035]

[0036] Compound 5: As shown in S5;

[0037]

[0038] Compound 6: As shown in S6;

[0039]

[0040] Compound 7: As shown in S7;

[0041]

[0042] Compound 8: As shown in S8;

[0043]

[0044] Compound 9: As shown in S9;

[0045]

[0046] Compound 10: as shown in S10;

[0047]

[0048] Compound 11: as shown in S11;

[0049]

[0050] Compound 12: as shown in S12;

[0051]

[0052] Compound 13: as shown in S13;

[0053]

[0054] Compound 14: as shown in S14.

[0055] The compounds provided by the present invention also include pharmaceutically acceptable equivalents of the compound or mixtures of two or more of them.

[0056] Preferably, the compounds provided by the present invention may include one or a mixture of two or more of pharmaceutically acceptable salts, hydrates, solvates, metabolites, and prodrugs.

[0057] Preferably, the compounds provided by the present invention include acid salts or base salts of the compounds provided by the present invention. The pharmaceutically acceptable salts have the pharmaceutical activity of the compound and meet the requirements both biologically and in practical applications.

[0058] The present invention provides a pharmaceutical composition comprising a compound shown in Formula I, its stereoisomers, or its pharmaceutically acceptable salts, hydrates or solvates, and a pharmaceutically acceptable carrier.

[0059] The present invention provides the use of a compound shown in Formula I, its stereoisomers, or its pharmaceutically acceptable salts, hydrates or solvates in the preparation of a drug for treating cancer, tumor, autoimmune disease, neurodegenerative disease, metabolic disease or metastatic disease.

[0060] The present invention provides the use of a compound represented by formula I, its stereoisomers or its pharmaceutically acceptable salts, hydrates or solvates in the preparation of drugs for treating tenosynovial giant cell tumor, ovarian cancer, pancreatic cancer, prostate cancer, non-small cell lung cancer, breast cancer, renal cell carcinoma, liver cancer, cervical cancer, papillary thyroid cancer, colorectal cancer, gastrointestinal stromal tumor, melanoma, mesothelioma, osteosarcoma, head and neck cancer, glioblastoma, leukemia, peritoneal malignancy, peripheral T-cell lymphoma, bone metastatic cancer, multiple myeloma, metastasis of primary tumor site, myeloproliferative disease, hyperproliferative disease, metabolic disease, rheumatoid arthritis, rheumatoid arthritis, osteoarthritis, pigmented villonodular synovitis, multiple sclerosis, autoimmune nephritis, lupus, Crohn's disease, asthma, neurodegenerative disease, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, osteoporosis, hypereosinophilic syndrome, mastocytosis or mast cell leukemia;

[0061] Preferably, it is used in the preparation of drugs for treating tenosynovial giant cell tumor, multiple myeloma, glioblastoma, metastasis of primary tumor site or bone metastatic cancer, metabolic disease, graft-versus-host disease, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease. Detailed implementation mode

[0062] The present invention discloses azaindole compounds and their uses. Those skilled in the art can draw on the content of this article and appropriately modify the process parameters to achieve. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and those skilled in the art can obviously make changes or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0063] The following examples can enable those skilled in the art to understand the present invention more comprehensively, but do not limit the present invention in any way.

[0064] Example 1: Synthesis of Compound S1

[0065] Synthesis route:

[0066]

[0067] Synthesis process:

[0068] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. For post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0069] ESI-MS: 323.1[M+H]+

[0070] Step 2: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0071] ESI-MS: 475.1[M+H]+

[0072] Step 3: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, it was washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0073] ESI-MS: 259.0[M+H]+

[0074] Step 4: Take indoleacetic acid (0.5 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from Step 3 (0.62 g, 1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The reaction system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. 0.4 g of a pale yellow solid was obtained (yield 40%).

[0075] ESI-MS: 416.1[M+H]+

[0076] Step 5: Take the product from Step 4 (0.3 g, 1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after addition, heat to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The reaction system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purify by column chromatography. 57 mg of a pale yellow solid was obtained (yield 20%).

[0077] ESI-MS: 402.1[M+H]+

[0078] 1H NMR (400 MHz, DMSO-d6) δ 11.64 (s, 1H), 10.95 (s, 1H), 8.17 (d, J = 2.3 Hz, 1H), 8.02–7.94 (m, 2H), 7.44–7.35 (m, 2H), 7.32–7.24 (m, 2H), 6.99 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.92 (ddd, J = 8.1, 7.1, 1.2 Hz, 1H), 6.44 (t, J = 8.9 Hz, 2H), 6.18 (dd, J = 2.1, 1.0 Hz, 1H), 3.84 (s, 2H), 3.56 (td, J = 7.4, 5.6 Hz, 2H), 2.95 (t, J = 7.3 Hz, 2H).

[0079] Example 2: Synthesis of Compound S2

[0080] Synthesis route:

[0081]

[0082] Synthesis process:

[0083] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), and TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added. Then, (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added, and the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. After post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0084] ESI-MS: 323.1 [M+H]+

[0085] Step 2: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml). Finally, tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added, and the reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0086] ESI-MS: 475.1 [M+H]+

[0087] Step 3: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), and triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added. The reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, and it was washed once with saturated sodium bicarbonate and twice with water. After drying and concentrating, it was purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0088] ESI-MS: 259.0 [M+H]+

[0089] Step 4: Take 2-pyridineacetic acid (0.39 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (0.62 g, 1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The system was diluted with EA (100 ml), washed with saturated sodium bicarbonate 3 times, dried, concentrated, and purified by column chromatography. A pale yellow solid of 0.358 g (yield 40%) was obtained.

[0090] ESI-MS: 378.1 [M+H]+

[0091] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after adding, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried, and concentrated. Purify by column chromatography. A pale yellow solid of 52 mg (yield 20%) was obtained.

[0092] ESI-MS: 364.1 [M+H]+

[0093] 1 H NMR (400 MHz, DMSO-d6) δ 11.63 (s, 1H), 8.49 (ddd, J = 4.9, 1.9, 0.9 Hz, 1H), 8.16 (d, J = 2.3 Hz, 1H), 7.96 (dd, J = 9.4, 2.4 Hz, 2H), 7.68 (td, J = 7.6, 1.9 Hz, 1H), 7.36 (d, J = 2.5 Hz, 1H), 7.30–7.23 (m, 2H), 7.20 (ddd, J = 7.5, 4.8, 1.2 Hz, 1H), 6.39 (d, J = 8.5 Hz, 2H), 3.83 (s, 2H), 3.54 (q, J = 7.0, 5.4 Hz, 2H), 2.96 (t, J = 7.3 Hz, 2H).

[0094] Example 3: Synthesis of Compound S3

[0095] Synthesis route:

[0096]

[0097] Synthesis process:

[0098] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. For post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0099] ESI-MS: 323.1[M+H]+

[0100] Step 2: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0101] ESI-MS: 475.1[M+H]+

[0102] Step 3: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, it was washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0103] ESI-MS: 259.0[M+H]+

[0104] Step 4: Take methyl N-methylpyrrolidine-2-acetate (0.43 g, 1.2 eq, 2.86 mmol), dissolve it in THF (10 ml), add the pure product from Step 3 (0.62 g, 1 eq, 2.38 mmol), cool the system to 0 °C, and finally add dropwise NaHMDS (1.2 eq, 2.86 mmol). After addition, let it warm to room temperature and react for 12 h. TLC (DCM:ME = 15:1, Rf of starting material = 0.2, Rf of product = 0.35), there was still a large amount of starting material remaining. Pour the system into 60 ml of water, extract 4 times with EA (40 ml), combine the organic phases, dry and concentrate, and then purify by column chromatography. Obtain 0.36 g of a pale yellow solid (yield 40%).

[0105] ESI-MS: 380.1 [M+H]+

[0106] Step 5: Take the product from Step 4 (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it 3 times, and after addition, heat to 60 °C and react. TLC (DCM:ME = 15:1, Rf of starting material = 0.35, Rf of product = 0.3), the reaction was complete. Pour the system into 50 ml of water, extract 4 times with EA (30 ml), combine the organic phases, dry and concentrate. Purify by column chromatography. Obtain 53 mg of a pale yellow solid (yield 20%).

[0107] ESI-MS: 366.1 [M+H]+

[0108] 1 H NMR (400 MHz, DMSO-d6) δ 11.63 (s, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.96 (dd, J = 12.8, 2.4 Hz, 2H), 7.37 (d, J = 2.4 Hz, 1H), 7.27 (dd, J = 8.7, 2.4 Hz, 1H), 6.59 (t, J = 2.3 Hz, 1H), 6.40 (t, J = 7.6 Hz, 2H), 5.96–5.74 (m, 2H), 3.83 (s, 2H), 3.51 (s, 3H), 3.42–3.38 (m, 2H), 2.73 (t, J = 7.5 Hz, 2H).

[0109] Example 4: Synthesis of Compound S4

[0110] Synthesis route:

[0111]

[0112] Synthesis process:

[0113] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), and TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added. Then, (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added, and the mixture was reacted at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of the starting material = 0.3, Rf of the product = 0.6), the reaction was complete. After workup, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0114] ESI-MS: 323.1[M+H]+

[0115] Step 2: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml). Finally, tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added, and the mixture was reacted at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of the starting indole = 0.5, Rf of the starting aldehyde = 0.95, Rf of the product = 0.3), the indole reaction was complete. The system was diluted with EA (150 ml), washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0116] ESI-MS: 475.1[M+H]+

[0117] Step 3: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), and triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added. The mixture was reacted at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of the starting material = 0.3, Rf of the product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, and the mixture was washed once with saturated sodium bicarbonate and twice with water. After drying and concentration, it was purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0118] ESI-MS: 259.0[M+H]+

[0119] Step 4: Take 2-pyrroleacetic acid (0.36 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (0.62 g, 1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The reaction system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. Obtained 0.347 g of a pale yellow solid (yield 40%).

[0120] ESI-MS: 366.1 [M+H]+

[0121] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after adding, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The reaction system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purified by column chromatography. Obtained 50 mg of a pale yellow solid (yield 20%).

[0122] ESI-MS: 352.1 [M+H]+

[0123] 1 1H NMR (400 MHz, DMSO-d6) δ 11.63 (s, 1H), 10.40 (s, 1H) 8.16 (d, J = 2.4 Hz, 1H), 7.96 (dd, J = 12.8, 2.4 Hz, 2H), 7.37 (d, J = 2.4 Hz, 1H), 7.27 (dd, J = 8.7, 2.4 Hz, 1H), 6.59 (t, J = 2.3 Hz, 1H), 6.40 (t, J = 7.6 Hz, 2H), 5.96–5.74 (m, 2H), 3.83 (s, 2H), 3.42–3.38 (m, 2H), 2.73 (t, J = 7.5 Hz, 2H).

[0124] Example 5: Synthesis of Compound S5

[0125] Synthesis route:

[0126]

[0127] Synthesis process:

[0128] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. For post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0129] ESI-MS: 323.1[M+H]+

[0130] Step 2: Fluoroazaindole (1 eq, 0.84 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.56 g of brown solid was obtained (yield 85%).

[0131] ESI-MS: 459.2[M+H]+

[0132] Step 3: The pure product of the second step (1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.21 g of yellow solid was obtained (yield 95%).

[0133] ESI-MS: 243.1[M+H]+

[0134] Step 4: Take indoleacetic acid (0.5 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, Rf of the starting material = 0.2, Rf of the product = 0.35), and there was still a large amount of starting material remaining. The reaction system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. 0.38 g of pale yellow solid was obtained (yield 40%).

[0135] ESI-MS: 400.1 [M+H]+

[0136] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after addition, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, Rf of the starting material = 0.35, Rf of the product = 0.3), and the reaction was complete. The reaction system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purify by column chromatography. 55 mg of pale yellow solid was obtained (yield 20%).

[0137] ESI-MS: 386.1 [M+H]+

[0138] 1 1H NMR (400 MHz, DMSO-d6) δ 11.52 (s, 1H), 10.93 (s, 1H), 8.15 (dd, J = 2.7, 1.7 Hz, 1H), 8.00 (d, J = 2.4 Hz, 1H), 7.74 (dd, J = 9.7, 2.8 Hz, 1H), 7.44–7.34 (m, 2H), 7.28 (ddd, J = 7.4, 5.9, 1.7 Hz, 2H), 6.99 (ddd, J = 8.2, 7.1, 1.3 Hz, 1H), 6.94–6.89 (m, 1H), 6.41 (dd, J = 6.9, 4.7 Hz, 2H), 6.18 (t, J = 1.4 Hz, 1H), 3.83 (s, 2H), 3.55 (q, J = 7.0 Hz, 2H), 2.95 (t, J = 7.3 Hz, 2H).

[0139] Example 6: Synthesis of Compound S6

[0140] Synthesis route:

[0141]

[0142] Synthesis process:

[0143] The first step: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, and then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. For post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0144] ESI-MS: 323.1 [M+H]+

[0145] The second step: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0146] ESI-MS: 475.1 [M+H]+

[0147] The third step: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, it was washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0148] ESI-MS: 259.0 [M+H]+

[0149] Step 4: Take 5-chloro-2-pyridineacetic acid (0.49 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (0.62 g, 1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, Rf of the raw material = 0.2, Rf of the product = 0.35), and there was still a large amount of raw material remaining. The system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. Obtained 0.39 g of a pale yellow solid (yield 40%).

[0150] ESI-MS: 412.0 [M+H]+

[0151] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it 3 times, and after adding, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, Rf of the raw material = 0.35, Rf of the product = 0.3), and the reaction was complete. The system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purified by column chromatography. Obtained 57 mg of a pale yellow solid (yield 20%).

[0152] ESI-MS: 398.1 [M+H]+

[0153] 1 H NMR (400 MHz, DMSO-d6) δ 11.62 (s, 1H), 8.53 (d, J = 2.6 Hz, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.96 (dd, J = 9.0, 2.4 Hz, 2H), 7.81 (dd, J = 8.3, 2.6 Hz, 1H), 7.36 (d, J = 2.4 Hz, 1H), 7.31 (d, J = 8.4 Hz, 1H), 7.26 (dd, J = 8.5, 2.5 Hz, 1H), 6.40–6.33 (m, 2H), 3.83 (s, 2H), 3.56–3.51 (m, 2H), 2.97 (d, J = 7.1 Hz, 2H).

[0154] Example 7: Synthesis of Compound S7

[0155] Synthesis route:

[0156]

[0157] Synthesis process:

[0158] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), and TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added. Then, (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added, and the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. After post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0159] ESI-MS: 323.1 [M+H]+

[0160] Step 2: Fluoroazaindole (1 eq, 0.84 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml). Finally, tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added, and the reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. The system was diluted with EA (150 ml), washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.56 g of brown solid was obtained (yield 85%).

[0161] ESI-MS: 459.2 [M+H]+

[0162] Step 3: The pure product of the second step (1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), and triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added. The reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.21 g of yellow solid was obtained (yield 95%).

[0163] ESI-MS: 243.1 [M+H]+

[0164] Step 4: Take 5-chloro-2-pyridineacetic acid (0.49 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. A pale yellow solid of 0.376 g (yield 40%) was obtained.

[0165] ESI-MS: 396.1[M+H]+

[0166] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it 3 times, and after adding, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purify by column chromatography. A pale yellow solid of 54 mg (yield 20%) was obtained.

[0167] ESI-MS: 382.1[M+H]+

[0168] 1 H NMR (400 MHz, DMSO-d6) δ 11.52 (s, 1H), 8.53 (d, J = 2.6 Hz, 1H), 8.15 (s, 1H), 7.97 (s, 1H), 7.81 (dd, J = 8.5, 2.6 Hz, 1H), 7.73 (dd, J = 9.5, 2.7 Hz, 1H), 7.36 (s, 1H), 7.31 (d, J = 8.3 Hz, 1H), 7.27 (dd, J = 8.5, 2.4 Hz, 1H), 6.37 (d, J = 8.1 Hz, 2H), 3.82 (s, 2H), 3.54 (q, J = 6.8 Hz, 2H), 2.96 (t, J = 7.1 Hz, 2H).

[0169] Example 8: Synthesis of Compound S8

[0170] Synthesis route:

[0171]

[0172] Synthesis process:

[0173] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, and then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. The reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. After work-up, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0174] ESI-MS: 323.1[M+H]+

[0175] Step 2: Chlorinated aza-indole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. The system was diluted with EA (150 ml), washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0176] ESI-MS: 475.1[M+H]+

[0177] Step 3: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0178] ESI-MS: 259.0[M+H]+

[0179] Step 4: Take 2-pyridinepropionic acid (0.43 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (0.62 g, 1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. Obtained 0.372 g of a pale yellow solid (yield 40%).

[0180] ESI-MS: 392.1 [M+H]+

[0181] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after adding, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purified by column chromatography. Obtained 54 mg of a pale yellow solid (yield 20%).

[0182] ESI-MS: 378.1 [M+H]+

[0183] 1 1H NMR (400 MHz, DMSO-d6) δ 11.61 (s, 1H), 8.50–8.43 (m, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.94 (d, J = 2.1 Hz, 2H), 7.68 (td, J = 7.6, 1.9 Hz, 1H), 7.35 (d, J = 2.4 Hz, 1H), 7.26 (dd, J = 8.2, 2.8 Hz, 2H), 7.21–7.15 (m, 1H), 6.38 (d, J = 8.5 Hz, 2H), 3.82 (s, 2H), 3.22 (t, J = 6.3 Hz, 2H), 2.80–2.75 (m, 2H), 2.24 (t, J = 7.5 Hz, 1H), 2.15 (t, J = 7.5 Hz, 1H).

[0184] Example 9: Synthesis of Compound S9

[0185] Synthesis route:

[0186]

[0187] Synthesis process:

[0188] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, and then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. For post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0189] ESI-MS: 323.1 [M+H]+

[0190] Step 2: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0191] ESI-MS: 475.1 [M+H]+

[0192] Step 3: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0193] ESI-MS: 259.0 [M+H]+

[0194] Step 4: Take cyclopropyl-2-pyridineacetic acid (0.466 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (0.62 g, 1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. Dilute the system with EA (100 ml), wash it with saturated sodium bicarbonate three times, dry and concentrate, and then purify by column chromatography. Obtain 0.383 g of a pale yellow solid (yield 40%).

[0195] ESI-MS: 404.1[M+H]+

[0196] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it three times, and after addition, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. Pour the system into 50 ml of water, extract it four times with EA (30 ml), combine the organic phases, dry and concentrate. Purify by column chromatography. Obtain 56 mg of a pale yellow solid (yield 20%).

[0197] ESI-MS: 390.1[M+H]+

[0198] 1 1H NMR (400 MHz, DMSO-d6) δ 11.63 (s, 1H), 8.44 (ddd, J = 4.9, 1.9, 0.9 Hz, 1H), 8.16 (d, J = 2.3 Hz, 1H), 7.97 (dd, J = 10.8, 2.3 Hz, 2H), 7.63 (td, J = 7.7, 1.9 Hz, 1H), 7.36 (dt, J = 8.1, 1.4 Hz, 2H), 7.25 (dd, J = 8.5, 2.4 Hz, 1H), 7.12 (ddd, J = 7.5, 4.8, 1.1 Hz, 1H), 6.46 (d, J = 8.5 Hz, 1H), 6.36 (s, 1H), 3.83 (s, 2H), 3.71 (d, J = 4.4 Hz, 2H), 1.13 (q, J = 3.7 Hz, 2H), 0.99 (q, J = 3.7 Hz, 2H).

[0199] Example 10: Synthesis of Compound S10

[0200] Synthesis route:

[0201]

[0202] Synthesis process:

[0203] First step: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, and then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. After post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0204] ESI-MS: 323.1 [M+H]+

[0205] Second step: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.5 g of brown solid was obtained (yield 85%).

[0206] ESI-MS: 475.1 [M+H]+

[0207] Third step: The pure product of the second step (2.5 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane hydride (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, it was washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.3 g of yellow solid was obtained (yield 95%).

[0208] ESI-MS: 259.0 [M+H]+

[0209] Step 4: Take difluoro-2-pyridineacetic acid (0.49 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (0.62 g, 1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. Obtained 0.39 g of a pale yellow solid (yield 40%).

[0210] ESI-MS: 414.1 [M+H]+

[0211] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after addition, heat to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purified by column chromatography. Obtained 57 mg of a pale yellow solid (yield 20%).

[0212] ESI-MS: 400.1 [M+H]+

[0213] 1 H NMR (400 MHz, DMSO-d6) δ 11.63 (s, 1H), 8.70–8.66 (m, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.99–7.88 (m, 3H), 7.68 (d, J = 7.8 Hz, 1H), 7.52 (dd, J = 7.4, 4.9 Hz, 1H), 7.34 (d, J = 2.4 Hz, 1H), 7.27 (dd, J = 8.6, 2.4 Hz, 1H), 6.69–6.63 (m, 1H), 6.49 (d, J = 8.5 Hz, 1H), 4.21 (td, J = 15.4, 6.4 Hz, 2H), 3.82 (s, 2H).

[0214] Example 11: Synthesis of Compound S11

[0215] Synthesis route:

[0216]

[0217] Synthesis process:

[0218] Step 1: 2-Amino-5-formyl-6-fluoro-pyridine (1 eq, 2.3 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. For post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.45 g of white solid was obtained (yield 80%).

[0219] ESI-MS: 341.1[M+H]+

[0220] Step 2: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2.11 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water three times, dried and concentrated, and the crude product was purified by column chromatography. 2.6 g of brown solid was obtained (yield 85%).

[0221] ESI-MS: 493.1[M+H]+

[0222] Step 3: The pure product of the second step (2.6 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, it was washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.38 g of yellow solid was obtained (yield 95%).

[0223] ESI-MS: 277.0[M+H]+

[0224] Step 4: Take cyclopropyl-2-pyridineacetic acid (0.466 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The reaction system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. Obtained 0.4 g of a pale yellow solid (yield 40%).

[0225] ESI-MS: 422.1[M+H]+

[0226] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it 3 times, and after adding, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The reaction system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purified by column chromatography. Obtained 58 mg of a pale yellow solid (yield 20%).

[0227] ESI-MS: 408.1[M+H]+

[0228] 1 H NMR (400 MHz, DMSO-d6) δ 11.66 (s, 1H), 8.47–8.41 (m, 1H), 8.18 (d, J = 2.3 Hz, 1H), 7.94 (d, J = 2.3 Hz, 1H), 7.65 (td, J = 7.7, 1.9 Hz, 1H), 7.39 (dd, J = 10.6, 8.1 Hz, 1H), 7.34–7.28 (m, 2H), 7.13 (dd, J = 7.4, 4.7 Hz, 1H), 6.77 (s, 1H), 6.36 (dd, J = 8.2, 1.7 Hz, 1H), 3.82 (s, 2H), 3.64 (d, J = 5.4 Hz, 2H), 1.14 (q, J = 3.7 Hz, 2H), 1.00 (q, J = 3.7 Hz, 2H).

[0229] Example 12: Synthesis of Compound S12

[0230] Synthesis route:

[0231]

[0232] Synthesis process:

[0233] First step: 2-Amino-5-formyl-6-fluoropyridine (1 eq, 2.3 g, 16.38 mmol) was dissolved in THF (40 ml), TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added, and then (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added. After addition, the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of raw material = 0.3, Rf of product = 0.6), the reaction was complete. For post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.45 g of white solid was obtained (yield 80%).

[0234] ESI-MS: 341.1 [M+H]+

[0235] Second step: Chlorinated azaindole (1 eq, 0.94 g, 6.21 mmol) and the product of the first step (1 eq, 2.11 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml), and finally tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added. The reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of raw material indole = 0.5, Rf of raw material aldehyde = 0.95, Rf of product = 0.3), the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water 3 times, dried and concentrated, and the crude product was purified by column chromatography. 2.6 g of brown solid was obtained (yield 85%).

[0236] ESI-MS: 493.1 [M+H]+

[0237] Third step: The pure product of the second step (2.6 g, 1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added, and the reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of raw material = 0.3, Rf of product = 0.2), the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, it was washed once with saturated sodium bicarbonate and twice with water, dried and concentrated, and then purified by column chromatography. 1.38 g of yellow solid was obtained (yield 95%).

[0238] ESI-MS: 277.0 [M+H]+

[0239] Step 4: Take difluoro-2-pyridineacetic acid (0.49 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The reaction system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. Obtained 0.41 g of a pale yellow solid (yield 40%).

[0240] ESI-MS: 432.0 [M+H]+

[0241] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after addition, heat to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The reaction system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purified by column chromatography. Obtained 60 mg of a pale yellow solid (yield 20%).

[0242] ESI-MS: 418.1 [M+H]+

[0243] 1 H NMR (400 MHz, DMSO-d6) δ 11.67 (s, 1H), 8.69 (d, J = 4.9 Hz, 1H), 8.18 (d, J = 2.3 Hz, 1H), 7.99–7.91 (m, 2H), 7.68 (d, J = 7.9 Hz, 1H), 7.53 (dd, J = 7.6, 4.8 Hz, 1H), 7.43 (dd, J = 10.6, 8.1 Hz, 1H), 7.30 (d, J = 2.4 Hz, 1H), 7.10 (t, J = 6.5 Hz, 1H), 6.40 (dd, J = 8.2, 1.6 Hz, 1H), 4.14 (td, J = 15.2, 6.4 Hz, 2H), 3.82 (s, 2H).

[0244] Example 13: Synthesis of Compound S13

[0245] Synthesis route:

[0246]

[0247] Synthesis process:

[0248] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), and TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added. Then, (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added, and the reaction was carried out at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of the raw material = 0.3, Rf of the product = 0.6), and the reaction was complete. After post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0249] ESI-MS: 323.1 [M+H]+

[0250] Step 2: Trifluoromethylazaindole (1.15 eq, 0.84 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml). Finally, tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added, and the reaction was carried out at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of the raw material indole = 0.5, Rf of the raw material aldehyde = 0.95, Rf of the product = 0.3), and the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water three times, dried and concentrated, and the crude product was purified by column chromatography. 2.68 g of brown solid was obtained (yield 85%).

[0251] ESI-MS: 509.2 [M+H]+

[0252] Step 3: The pure product of the second step (1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), and triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added. The reaction was carried out at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of the raw material = 0.3, Rf of the product = 0.2), and the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, and it was washed once with saturated sodium bicarbonate and twice with water. After drying and concentrating, it was purified by column chromatography. 1.47 g of yellow solid was obtained (yield 95%).

[0253] ESI-MS: 293.1 [M+H]+

[0254] Step 4: Take difluoro-2-pyridineacetic acid (0.49 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. The reaction system was diluted with EA (100 ml), washed 3 times with saturated sodium bicarbonate, dried and concentrated, and then purified by column chromatography. 0.425 g of a pale yellow solid was obtained (yield 40%).

[0255] ESI-MS: 448.1[M+H]+

[0256] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it in 3 portions, and after addition, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. The reaction system was poured into 50 ml of water, extracted 4 times with EA (30 ml), the organic phases were combined, dried and concentrated. Purify by column chromatography. 62 mg of a pale yellow solid was obtained (yield 20%).

[0257] ESI-MS: 434.1[M+H]+

[0258] 1 H NMR (400 MHz, DMSO-d6) δ 11.94 (s, 1H), 8.68 (dd, J = 4.5, 1.4 Hz, 1H), 8.52 (d, J = 2.1 Hz, 1H), 8.25 (d, J = 2.2 Hz, 1H), 7.95 (td, J = 3.6, 1.8 Hz, 2H), 7.67 (dt, J = 7.9, 1.1 Hz, 1H), 7.56–7.48 (m, 1H), 7.44 (d, J = 2.4 Hz, 1H), 7.30 (dd, J = 8.5, 2.4 Hz, 1H), 6.65 (t, J = 6.4 Hz, 1H), 6.52–6.48 (m, 1H), 4.21 (td, J = 15.4, 6.4 Hz, 2H), 3.91 (s, 2H).

[0259] Example 14: Synthesis of Compound S14

[0260] Synthesis route:

[0261]

[0262] Synthesis process:

[0263] Step 1: 2-Amino-5-formylpyridine (1 eq, 2 g, 16.38 mmol) was dissolved in THF (40 ml), and TEA (3 eq, 4.97 g, 49.14 mmol) and DMAP (0.2 eq, 0.4 g, 3.28 mmol) were added. Then, (Boc)2O (2.3 eq, 8.22 g, 37.67 mmol) was added, and the mixture was reacted at room temperature for 12 h. TLC (PE:EA = 5:1, Rf of the raw material = 0.3, Rf of the product = 0.6), and the reaction was complete. After post-treatment, the system was directly concentrated to dryness and purified by column chromatography. 4.3 g of white solid was obtained (yield 80%).

[0264] ESI-MS: 323.1 [M+H]+

[0265] Step 2: Trifluoromethylazaindole (1.15 eq, 0.84 g, 6.21 mmol) and the product of the first step (1 eq, 2 g, 6.21 mmol) were added to isopropanol (20 ml) and water (4 ml). Finally, tetramethylguanidine (0.5 eq, 0.36 g, 3.11 mmol) was added, and the mixture was reacted at 25 °C for 12 h. TLC (DCM:ME = 15:1, Rf of the raw material indole = 0.5, Rf of the raw material aldehyde = 0.95, Rf of the product = 0.3), and the indole reaction was complete. After the system was diluted with EA (150 ml), it was washed with water three times, dried and concentrated, and the crude product was purified by column chromatography. 2.68 g of brown solid was obtained (yield 85%).

[0266] ESI-MS: 509.2 [M+H]+

[0267] Step 3: The pure product of the second step (1 eq, 5.28 mmol) was added to acetonitrile (25 ml) and trifluoroacetic acid (5 ml), and triethylsilane (2.46 g, 4 eq, 21.12 mmol) was added. The mixture was reacted at 60 °C for 20 h. TLC (DCM:ME = 15:1, Rf of the raw material = 0.3, Rf of the product = 0.2), and the reaction was complete. After the system was cooled to room temperature, the reaction solution was concentrated. EA (200 ml) was added, and it was washed once with saturated sodium bicarbonate and twice with water. After drying and concentrating, it was purified by column chromatography. 1.47 g of yellow solid was obtained (yield 95%).

[0268] ESI-MS: 293.1 [M+H]+

[0269] Step 4: Take 5-chloro-2-pyridineacetic acid (0.49 g, 1.2 eq, 2.86 mmol), dissolve it in DMF (5 ml), add DIPEA (0.92 g, 3 eq, 7.14 mmol), then add TBTU (0.95 g, 1.25 eq, 2.97 mmol), stir for 2 min, and finally add the pure product from the third step (1 eq, 2.38 mmol). React at room temperature for 24 h. TLC (DCM:ME = 15:1, starting material Rf = 0.2, product Rf = 0.35), and there was still a large amount of starting material remaining. Dilute the system with EA (100 ml), wash it 3 times with saturated sodium bicarbonate, dry and concentrate, and then purify by column chromatography. Obtain 0.423 g of a pale yellow solid (yield 40%).

[0270] ESI-MS: 446.1 [M+H]+

[0271] Step 5: Take the product from the fourth step (1 eq, 0.72 mmol), dissolve it in THF (8 ml), add lithium aluminum hydride (55 mg, 2 eq, 1.44 mmol), add it 3 times, and after adding, heat up to 60 °C for reaction. TLC (DCM:ME = 15:1, starting material Rf = 0.35, product Rf = 0.3), and the reaction was complete. Pour the system into 50 ml of water, extract it 4 times with EA (30 ml), combine the organic phases, dry and concentrate. Purify by column chromatography. Obtain 62 mg of a pale yellow solid (yield 20%).

[0272] ESI-MS: 432.1 [M+H]+

[0273] 1 H NMR (400 MHz, DMSO-d6) δ 11.94 (s, 1H), 8.53 (q, J = 2.5 Hz, 2H), 8.25 (d, J = 2.1 Hz, 1H), 7.99 (d, J = 2.3 Hz, 1H), 7.80 (dd, J = 8.3, 2.6 Hz, 1H), 7.46 (d, J = 2.4 Hz, 1H), 7.30 (dd, J = 8.5, 6.9 Hz, 2H), 6.42–6.34 (m, 2H), 3.91 (s, 2H), 3.54 (q, J = 6.8 Hz, 2H), 2.96 (t, J = 7.1 Hz, 2H).

[0274] Enzymatic activity evaluation of the compound of Example 15

[0275] 1. In vitro biochemical kinase experiment of CSF1R

[0276] This invention uses ADP-Glo TMKinase Detection Kit (Promega, cat. No. V9101), which quantifies the amount of ADP generated during the kinase reaction to evaluate the kinase inhibitory activity of compounds. The specific experimental procedure is as follows: The kinase reaction is carried out in a 384-well plate (Greiner, cat. No. 784075), with each well containing 0.4 nM CSF1R, 100 μM ATP, and 0.1 mg / ml polypeptide; A kinase reaction system containing Hepes, MgCl2, Brij35, EGTA, and DTT is prepared for the preparation of 2×ATP and substrate solution as well as 2×kinase solution respectively; 20 nL of serially diluted compounds are transferred to the 384-well detection plate by Echo 655. 3 μL of 2×kinase solution is added thereto, mixed well, and incubated at 25 °C for 20 min; 3 μL of 2×substrate and ATP solution is added to the wells and incubated at 25 °C for 30 min; 4 μL of ADP-Glo reagent is added to the wells and incubated at 25 °C for 40 min; 6 μL of Kinase Detection Reagent is added to the wells and incubated at 25 °C for 40 min; The RLU (Relative luminescence unit) signal is read using a BMG multi-functional microplate reader. The signal intensity is used to characterize the degree of kinase activity, and the four-parameter curve in Graphpad Prism is used to determine the IC50 value of the compound.

[0277] 2. In Vitro Biochemical Kinase Assays for C-KIT / FLT3 / PDGFRα / PDGFRβ

[0278] Use the detection kit HTRF KinEASE-TK kit to detect the selectivity of compounds for tyrosine kinase targets (C-KIT, FLT3, PDGFRα, PDGFRβ). The specific experimental procedure is as follows: The kinase reaction carried out in this invention is performed in a 384-well plate (Greiner, cat. No. 784075), containing 1 nM of C-KIT, 6 μM of ATP, and 1 μM of TK substrate respectively; containing 0.024 nM of FLT3, 2 μM of ATP, and 1 μM of TK substrate; containing 0.12 nM of PDGFRα (Carna, cat. No. 08-157), 0.5 μM of ATP, and 1 μM of TK substrate; containing 0.23 nM of PDGFRβ, 1 μM of ATP, and 1 μM of TK substrate; prepare a 1× kinase reaction system containing MgCl2, MnCl, SEB, and DTT for preparing 2× ATP and substrate solution and 2× kinase solution; transfer 25 nL of the compound to the 384-well assay plate. Add 2.5 μL of 2× kinase solution, mix well, and incubate at 25 °C for 10 min; add 2.5 μL of 2× substrate and ATP solution to the wells and incubate at 25 °C for 30 min; prepare 2× XL665 and antibody solution with detection buffer; add 5 μL of Kinase Detection Reagent to the wells and incubate at 25 °C for 60 min; use a BMG multi-functional microplate reader to read the fluorescence signals at 620 nm (Cryptate) and 665 nm (XL665), and use the four-parameter curve in Graphpad Prism to determine the IC50 value of the compound. The results are shown in Table 1.

[0279] Example 16 Evaluation of the Cellular Activity of Compounds

[0280] Mouse myeloid leukemia lymphocyte line M-NFS-60 (RPMI1640 + 0.05 mM β-mercaptoethanol + 62 ng / mL CSF-1 + 10% FBS), human monocytic leukemia cell line THP-1 (RPMI1640 + 10% FBS + 0.05 mM β-mercaptoethanol + 1% P / S), mouse microglial cell line BV2 (MEM + 1% NEAA + 10% FBS + 1% P / S) were all purchased from Wuhan Punosai Life Science Co., Ltd. and maintained normal growth under the culture conditions of 37 °C and 5% CO2. Take BV2, M-NFS-60, and THP-1 cells in the logarithmic growth phase, and respectively at 8×10 per well 3 、3×10 4 、4×10 4Cells were seeded in 96-well plates, with each well containing 100 μL of medium, and three replicate wells were set for each drug concentration. After BV2 cells were seeded, they were cultured overnight at 37 °C and 5% CO2. After the cells adhered to the wall, CSF1R inhibitor was added and the cells were treated for 72 h. After the suspension cells M-NFS-60 and THP-1 were allowed to stand for an appropriate time, the drug could be directly added, and then the cells were incubated for 72 h. The Cell Titer-Glo Luminescent Cell Viability Assay (Promega, cat. No. G7571) was used

[0281] to evaluate cell viability. According to the reagent instructions, the CellTiter-Glo Buffer was thawed in advance before detection, and it was balanced to room temperature with the freeze-dried substrate. The two were mixed evenly to prepare the CellTiter-Glo Reagent detection reagent. At the same time, the cell culture plate and its contents were balanced at room temperature. 100 μL of CellTiter-Glo Reagent was added to the medium containing cells or the complete medium without cells, and a orbital shaker was used to mix for 2 min to induce cell lysis. Then the culture plate was incubated at room temperature for 10 min to stabilize the luminescence signal. The RLU (Relative luminescence unit) signal was read using a microplate reader. The signal intensity was used to characterize the cell viability, and the four-parameter curve in Graphpad Prism was used to determine the IC50 value of the compound. The results are shown in Table 1

[0282] Table 1 Detection results of compound enzyme activity and cell activity

[0283]

[0284] Note: "+" means the IC50 of the compound CSF1R kinase inhibitory activity > 30 nM; "++" means the IC50 of the compound CSF1R kinase inhibitory activity is 10 nM < IC50 < 30 nM; "+++" means the IC50 of the compound CSF1R kinase inhibitory activity < 10 nM

[0285] Example 17 Compound PK experiment

[0286] In the PK experiment of the compound drug in the embodiments of the present invention, male SD rats were used. One day before the experiment, the animals were fasted (about 12 h), and a single oral administration was adopted. The dose was 5 mg / Kg or 10 mg / kg, and the formulation was suspended in 0.5% CMC-Na. After the rats were administered, the blood sampling time points were 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 24 h after administration. All the above samples were placed in heparin sodium anticoagulated EP tubes and ice-bathed after collection, centrifuged at 4°C and 8000 rpm for 5 minutes, and the plasma was transferred to -20°C for storage and waiting for detection as soon as possible. The LC-MS / MS detection method that has been investigated for specificity, standard curve, precision, accuracy and dilution accuracy was used to determine the compound concentration in the collected samples. Winnonlin 5.2 was used to calculate the pharmacokinetic parameters. The PK data of the specific example compounds are shown in Tables 2 and 3 below.

[0287] Table 2 Compound PK parameters (i.g. 10.0 mg / kg)

[0288] Serial number <![CDATA[t 1 / 2 (h)]]> <![CDATA[T max (h)]]> <![CDATA[C max (ng / ml)]]> AUC (h*ng / ml) F(％) S2 2.74 0.50 5120 10426 36.3％ S3 2.00 0.50 818 1935 26.4％ S4 1.15 0.58 954 1434 21.7％ S5 1.31 1.00 305 761 11.8％

[0289] Table 3 Compound PK parameters (i.g. 5.0 mg / kg)

[0290] Serial number <![CDATA[t 1 / 2 (h)]]> <![CDATA[T max (h)]]> <![CDATA[C max (ng / ml)]]> AUC (h*ng / ml) F(％) S6 3.00 1.08 5953 18561 43.0％ S7 1.16 1.33 8640 24968 27.9％ S8 1.20 0.42 772 1224 28.6％ S9 2.17 0.83 881 2135 28.3％ S10 2.48 1.00 4840 14844 59.4％

Claims

1. A class of azaindole compounds represented by formula I, their stereoisomers, or pharmaceutically acceptable salts, hydrates or solvates thereof, characterized in that, wherein, R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, difluoromethyl or C1-C6 alkyl; R2 is selected from hydrogen, chlorine or fluorine; R3 and R4 are each independently selected from hydrogen, fluorine, C1-C3 alkyl, or R3 and R4 together with the connected carbon atom form an alkyl ring; A is selected from indole, substituted indole, pyridine, substituted pyridine, pyrrole, substituted pyrrole, pyrimidine or substituted pyrimidine; n is selected from the numbers 0, 1, 2 or 3.

2. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, characterized in that, wherein, R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl or C1-C3 alkyl; R2 is selected from hydrogen, chlorine or fluorine; R3 and R4 are each independently selected from hydrogen, fluorine, C1-C3 alkyl, or R3, R4 and the connected carbon atom together form an alkyl three-membered ring or a four-membered ring; A is selected from indole, substituted indole, pyridine, substituted pyridine, pyrrole or substituted pyrrole; n is selected from the numbers 0, 1 or 2.

3. The compound according to claim 1, and its stereoisomers or pharmaceutically acceptable salts thereof, characterized in that, wherein, R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl or methyl; R2 is selected from hydrogen or fluorine; R3 and R4 are both hydrogen at the same time, or R3 and R4 are both fluorine at the same time, or R3, R4 and the connected carbon atom together form an alkyl three-membered ring; A is selected from indole, pyridine, halogen-substituted pyridine, pyrrole or N-methylpyrrole; n is selected from the numbers 0 or 1.

4. The compound according to claim 1, and its stereoisomers or pharmaceutically acceptable salts thereof, characterized in that, The compounds are: Compound 1: as shown in S1; Compound 2: as shown in S2; Compound 3: as shown in S3; Compound 4: as shown in S4; Compound 5: as shown in S5; Compound 6: as shown in S6; Compound 7: as shown in S7; Compound 8: as shown in S8; Compound 9: as shown in S9; Compound 10: as shown in S10; Compound 11: as shown in S11; Compound 12: as shown in S12; Compound 13: as shown in S13; Compound 14: as shown in S14.

5. Use of the compound, its stereoisomer or its pharmaceutically acceptable salt according to any one of claims 1 to 4 in the treatment of cancer, tumor, autoimmune disease, neurodegenerative disease, metabolic disease or metastatic disease.

6. Use of the compound, its stereoisomer or its pharmaceutically acceptable salt according to any one of claims 1 to 4 in the preparation of drugs for treating giant cell tumor of tendon sheath, ovarian cancer, pancreatic cancer, prostate cancer, non-small cell lung cancer, breast cancer, renal cell carcinoma, liver cancer, cervical cancer, papillary thyroid cancer, colorectal cancer, gastrointestinal stromal tumor, melanoma, mesothelioma, osteosarcoma, head and neck cancer, glioblastoma, leukemia, peritoneal malignancy, peripheral T cell lymphoma, bone metastatic cancer, multiple myeloma, metastasis of primary tumor site, myeloproliferative disease, hyperproliferative disease, metabolic disease, rheumatoid arthritis, rheumatoid arthritis, osteoarthritis, pigmented villonodular synovitis, multiple sclerosis, autoimmune nephritis, lupus, Crohn's disease, asthma, neurodegenerative disease, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, osteoporosis, hypereosinophilic syndrome, mastocytosis or mast cell leukemia; Specifically, it is used in the preparation of drugs for treating giant cell tumor of tendon sheath, multiple myeloma, glioblastoma, metastasis of primary tumor site or bone metastatic cancer, metabolic diseases, graft-versus-host disease, amyotrophic lateral sclerosis, Alzheimer's disease, and Parkinson's disease.

7. A pharmaceutical composition, characterized in that, It comprises the compound or its pharmaceutically acceptable salt as described in any one of claims 1 to 4 and a pharmaceutically acceptable carrier.

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