Class of pyridine derivative compounds and use thereof

Abstract

A class of pyridine derivative compounds and a use thereof are provided, belonging to the field of pharmaceuticals. The compounds have CSF1R inhibitory activity and can be widely used in drugs for treating cancer, tumor, autoimmune disease, neurodegenerative disease, traumatic brain injury, metabolic disease, or metastatic disease.

Description

A class of pyridine-derived compounds and their uses

[0001] This application claims priority to Chinese Patent Application No. 202510055924.X, filed on January 14, 2025, entitled "A Class of Pyridine Derivative Compounds and Their Uses", the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of pharmaceutical technology, specifically relating to a class of pyridine-derived compounds and their uses. Background Technology

[0003] CSF-1R is a transmembrane receptor for tyrosine kinases and belongs to the CSF-1 / platelet-derived growth factor (PDGF) receptor family. This receptor family possesses 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, and monocytes). IL-34 has similar biological characteristics to CSF-1, but their main differences stem from different spatiotemporal regulation. Furthermore, CSF-1 exerts its effects through autocrine and paracrine pathways, while IL-34 acts only locally. Small molecule inhibitors or antibodies targeting CSF-1R have the activity of inhibiting the survival and proliferation of microglia, inflammatory macrophages, and tumor-associated macrophages (TAMs), thereby exerting anti-tumor, immunomodulatory, and neuroinflammatory-allergic effects. They are currently being used in drug development for various cancers, central nervous system degenerative diseases, and chronic graft-versus-host disease.

[0004] High levels of CSF-1 have been reported in breast cancer, pancreatic cancer, prostate cancer, ovarian cancer, kidney cancer, and many other types of cancer. 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 modulate 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 CSF-1R inhibitors can inhibit and kill overactivated microglia in the nervous system with high affinity and selectivity, effectively blocking the neuroinflammatory response driving disease progression, slowing nerve damage, and promoting nerve cell repair. In addition to their important roles in oncology and central nervous system degenerative diseases, CSF-1R inhibitors also have broad application prospects, covering multiple fields such as inflammation, metabolism, and bone-related diseases. Therefore, drug development targeting CSF-1R has become a research hotspot for scholars and pharmaceutical companies both domestically and internationally.

[0005] Compared to earlier approved CSF-1R multi-target inhibitors, such as Surufatinib, Pexidartinib, and Vorolanib, next-generation CSF-1R inhibitors need to possess better selectivity to reduce off-target effects and side effects, while also requiring further optimization of the pharmacokinetic properties of small molecule drugs. This invention aims to discover highly active and selective CSF1R inhibitors through structural modification of Pexidartinib. Summary of the Invention

[0006] Technical problem solved: This invention provides a class of pyridine-derived compounds with CSF1R inhibitory activity, which can be widely used as drugs for treating cancer, tumors, autoimmune diseases, graft-versus-host disease, neurodegenerative diseases, traumatic brain injury, metabolic diseases or metastatic diseases.

[0007] Technical solution: A class of compounds as shown in Formula I and Formula II, their stereoisomers, or their pharmaceutically acceptable salts, hydrates, or solvates, as shown in Formula I and Formula II.

[0008] R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, difluoromethyl, methoxy, morpholino, N-methylpiperazine, or N-methylpyrazole;

[0009] R2 is selected from hydrogen, chlorine, fluorine, C1-C3 alkyl, or halogen-substituted C1-C3 alkyl;

[0010] R3 is selected from hydrogen, C1-C6 alkyl, C3-C5 cycloalkyl, deuterated methyl or halogen-substituted C1-C6 alkyl;

[0011] R4 and R5 are each independently selected from hydrogen or C1-C6 alkyl groups, or R4 and R5 together with the carbon atoms they are connected to form C3-C6 cycloalkyl groups.

[0012] Preferably, R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, morpholino, N-methylpiperazine, or N-methylpyrazole;

[0013] R2 is selected from hydrogen, fluorine, or C1-C3 alkyl groups;

[0014] R3 is selected from hydrogen, C1-C3 alkyl, C3-C5 cycloalkyl, deuterated methyl or fluorine-substituted C1-C3 alkyl;

[0015] R4 and R5 are each independently selected from hydrogen, C1 to C3 alkyl groups, or R4 and R5 together with the carbon atoms they are connected to form C3 to C5 cycloalkyl groups.

[0016] Preferably, R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, morpholino, N-methylpiperazine, or N-methylpyrazole;

[0017] R2 is selected from hydrogen, fluorine, or methyl;

[0018] R3 is selected from hydrogen, C1-C3 alkyl, cyclopropyl or deuterated methyl;

[0019] R4 and R5 are each independently selected from hydrogen and methyl, or R4 and R5 together with the carbon atom they are connected to form a C3-C4 cycloalkyl group.

[0020] Preferably, the compound is:

[0021] Compound 1: As shown in S1;

[0022] Compound 2: As shown in S2;

[0023] Compound 3: As shown in S3;

[0024] Compound 4: As shown in S4;

[0025] Compound 5: As shown in S5;

[0026] Compound 6: As shown in S6;

[0027] Compound 7: As shown in S7;

[0028] Compound 8: As shown in S8;

[0029] Compound 9: As shown in S9;

[0030] Compound 10: as shown in S10;

[0031] Compound 11: As shown in S11;

[0032] Compound 12: as shown in S12;

[0033] Compound 13: As shown in S13;

[0034] Compound 14: As shown in S14.

[0035] Preferably, the compound is used in the preparation of drugs for treating cancer.

[0036] Preferably, the compound is used in the preparation of drugs for treating tumors, autoimmune diseases, neurodegenerative diseases, traumatic brain injury, metabolic diseases, or metastatic diseases.

[0037] Preferably, the compound is used 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 carcinoma, colorectal cancer, gastrointestinal stromal tumors, melanoma, mesothelioma, osteosarcoma, head and neck cancer, glioblastoma, leukemia, peritoneal malignant tumors, peripheral T-cell lymphoma, bone metastatic cancer, multiple myeloma, metastasis of primary tumor sites, myeloproliferative disorders, hyperproliferative disorders, metabolic disorders, rheumatoid arthritis, osteoarthritis, pigmented villonodular synovitis, multiple sclerosis, autoimmune nephritis, lupus, Crohn's disease, asthma, neurodegenerative diseases, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, osteoporosis, hypereosinophilic syndrome, mastocytosis or mast cell leukemia, graft-versus-host disease, and traumatic brain injury.

[0038] Preferably, the compound is used in the preparation of drugs for treating tenosynovial giant cell tumor, multiple myeloma, glioblastoma, metastatic or bone metastatic cancers at primary tumor sites, metabolic diseases, graft-versus-host disease, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, and traumatic brain injury.

[0039] A pharmaceutical composition comprising any one of the compounds described above or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Beneficial effects:

[0040] This invention provides a class of pyridine-derived compounds with CSF1R inhibitory activity, which can be widely used in the treatment of cancer, tumors, autoimmune diseases, neurodegenerative diseases, metabolic diseases or metastatic diseases. Detailed Implementation

[0041] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0042] Example 1: Synthesis of compound S1

[0043] Synthesis of intermediate B1 in step 1

[0044] Al (2 g, 16.38 mmol) was dissolved in THF (40 mL), followed by the addition of TEA (4.97 g, 49.14 mmol) and DMAP (0.4 g, 3.28 mmol), then (Boc)₂O (8.22 g, 37.67 mmol). The reaction was allowed to proceed at room temperature for 12 h, and TLC analysis showed complete reaction. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 4.3 g of a white solid, with a yield of 80%. ESI-MS: 323.1 [M+H] + .

[0045] Synthesis of intermediate C1 in step 2

[0046] Chloroazine indole (0.94 g, 6.21 mmol) and B1 (2 g, 6.21 mmol) were added to isopropanol (20 mL) and water (4 mL), followed by tetramethylguanidine (0.36 g, 3.11 mmol). The reaction was carried out at 25°C for 12 h. TLC analysis showed the reaction was complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 2.5 g of a pale yellow solid, yield 85%. ESI-MS: 475.1 [M+H] + .

[0047] Synthesis of intermediate D1 in step 3

[0048] C1 (2.5 g, 5.28 mmol) was added to a mixed solution of acetonitrile (25 mL) and trifluoroacetic acid (5 mL), followed by triethylsilane (2.46 g, 21.12 mmol). The reaction was carried out at 60°C for 20 h. TLC analysis showed the reaction was complete. After cooling to room temperature, the reaction solution was concentrated. EA (200 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, rotary evaporated, and subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 1.3 g of a pale yellow solid, 95% yield. ESI-MS: 259.0 [M+H] + .

[0049] Step 4: Synthesis of final product S1

[0050] 2-O-1-methylnicotinic acid (0.5 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally D1 (0.61 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 0.5 g of a pale yellow solid, yield 54%. ESI-MS: 394.1 [M+H] + .

[0051] 1 H NMR (400MHz, DMSO-d6) δ12.45(s,1H),11.71(s,1H),8.46(dd,J=7.3,2.2Hz,1H),8.35(d,J=2.3Hz,1H),8.23-8.13(m,3H), 8.01 (d, J = 2.3Hz, 1H), 7.73 (dd, J = 8.5, 2.4Hz, 1H), 7.44 (d, J = 2.4Hz, 1H), 6.59 (t, J = 6.9Hz, 1H), 4.03 (s, 2H), 3.62 (s, 3H).

[0052] Example 2: Synthesis of compound S2

[0053] Synthesis of intermediate B1 in step 1

[0054] Al (2 g, 16.38 mmol) was dissolved in THF (40 mL), followed by the addition of TEA (4.97 g, 49.14 mmol) and DMAP (0.4 g, 3.28 mmol), then (Boc)₂O (8.22 g, 37.67 mmol). The reaction was allowed to proceed at room temperature for 12 h, and TLC analysis showed complete reaction. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 4.3 g of a white solid, with a yield of 80%. ESI-MS: 323.1 [M+H] + .

[0055] Synthesis of intermediate C2 in step 2

[0056] Fluoroazaindole (0.94 g, 6.21 mmol) and B1 (2 g, 6.21 mmol) were added to isopropanol (20 mL) and water (4 mL), followed by tetramethylguanidine (0.36 g, 3.11 mmol). The reaction was carried out at 25°C for 12 h. TLC analysis showed the reaction was complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 2.5 g of a pale yellow solid, yield 85%. ESI-MS: 458.2 [M+H] + .

[0057] Synthesis of intermediate D2 in step 3

[0058] C2 (2.5 g, 5.28 mmol) was added to a mixed solution of acetonitrile (25 mL) and trifluoroacetic acid (5 mL), followed by triethylsilane (2.46 g, 21.12 mmol). The reaction was carried out at 60°C for 20 h. TLC analysis showed the reaction was complete. After cooling to room temperature, the reaction solution was concentrated. EA (200 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, rotary evaporated, and subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 1.3 g of a pale yellow solid, 95% yield. ESI-MS: 243.1 [M+H] + .

[0059] Step 4: Synthesis of final product S2

[0060] 2-O-1-methylnicotinic acid (0.5 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally D2 (0.58 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 0.48 g of a pale yellow solid, yield 54%. ESI-MS: 378.1 [M+H] + .

[0061] 1H NMR (400MHz, DMSO-d6) δ12.45(s,1H),11.60(s,1H),8.46(dd,J=7.4,2.2Hz,1H),8.40-8.30(m,1H),8.23-8.12(m,3 H),7.76(ddd,J=22.6,9.0,2.6Hz,2H),7.44(d,J=2.5Hz,1H),6.59(dd,J=7.4,6.5Hz,1H),4.02(s,2H),3.62(s,3H).

[0062] Example 3: Synthesis of compound S3

[0063] Synthesis of intermediate B1 in step 1

[0064] Al (2 g, 16.38 mmol) was dissolved in THF (40 mL), followed by the addition of TEA (4.97 g, 49.14 mmol) and DMAP (0.4 g, 3.28 mmol), then (Boc)₂O (8.22 g, 37.67 mmol). The reaction was allowed to proceed at room temperature for 12 h, and TLC analysis showed complete reaction. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 4.3 g of a white solid, with a yield of 80%. ESI-MS: 323.1 [M+H] + .

[0065] Synthesis of intermediate C2 in step 2

[0066] Fluoroazaindole (0.94 g, 6.21 mmol) and B1 (2 g, 6.21 mmol) were added to isopropanol (20 mL) and water (4 mL), followed by tetramethylguanidine (0.36 g, 3.11 mmol). The reaction was carried out at 25°C for 12 h. TLC analysis showed the reaction was complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 2.5 g of a pale yellow solid, yield 85%. ESI-MS: 458.2 [M+H] + .

[0067] Synthesis of intermediate D2 in step 3

[0068] C2 (2.5 g, 5.28 mmol) was added to a mixed solution of acetonitrile (25 mL) and trifluoroacetic acid (5 mL), followed by triethylsilane (2.46 g, 21.12 mmol). The reaction was carried out at 60°C for 20 h. TLC analysis showed the reaction was complete. After cooling to room temperature, the reaction solution was concentrated. EA (200 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, rotary evaporated, and subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 1.3 g of a pale yellow solid, 95% yield. ESI-MS: 243.1 [M+H] + .

[0069] Step 4: Synthesis of final product S3

[0070] 2-Hydroxy-3-pyridinecarboxylic acid (0.4 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally D2 (0.58 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 0.26 g of a pale yellow solid, with a yield of 30%. ESI-MS: 364.1 [M+H] + .

[0071] 1 H NMR (400MHz, DMSO-d6) δ12.78(s,1H),12.45(s,1H),11.60(s,1H),8.47(dd,J=7.3,2.2Hz,1H),8.34(d,J=2.3 Hz, 1H), 8.20-8.15 (m, 2H), 7.84-7.69 (m, 3H), 7.43 (d, J = 2.5Hz, 1H), 6.57 (dd, J = 7.2, 6.2Hz, 1H), 4.01 (s, 2H).

[0072] Example 4: Synthesis of compound S4

[0073] Synthesis of intermediate B1 in step 1

[0074] Al (2 g, 16.38 mmol) was dissolved in THF (40 mL), followed by the addition of TEA (4.97 g, 49.14 mmol) and DMAP (0.4 g, 3.28 mmol), then (Boc)₂O (8.22 g, 37.67 mmol). The reaction was allowed to proceed at room temperature for 12 h, and TLC analysis showed complete reaction. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 4.3 g of a white solid, with a yield of 80%. ESI-MS: 323.1 [M+H] + .

[0075] Synthesis of intermediate C2 in step 2

[0076] Fluoroazaindole (0.94 g, 6.21 mmol) and B1 (2 g, 6.21 mmol) were added to isopropanol (20 mL) and water (4 mL), followed by tetramethylguanidine (0.36 g, 3.11 mmol). The reaction was carried out at 25°C for 12 h. TLC analysis showed the reaction was complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 2.5 g of a pale yellow solid, yield 85%. ESI-MS: 458.2 [M+H] + .

[0077] Synthesis of intermediate D2 in step 3

[0078] C2 (2.5 g, 5.28 mmol) was added to a mixed solution of acetonitrile (25 mL) and trifluoroacetic acid (5 mL), followed by triethylsilane (2.46 g, 21.12 mmol). The reaction was carried out at 60°C for 20 h. TLC analysis showed the reaction was complete. After cooling to room temperature, the reaction solution was concentrated. EA (200 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, rotary evaporated, and subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 1.3 g of a pale yellow solid, 95% yield. ESI-MS: 243.1 [M+H] + .

[0079] Step 4: Synthesis of final product S4

[0080] 1-Cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid (0.51 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally D2 (0.58 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 0.43 g of a pale yellow solid, with a yield of 40%. ESI-MS: 404.1 [M+H] + .

[0081] 1 H NMR (400MHz, DMSO-d6) δ12.45(s,1H),11.61(s,1H),8.44(dd,J=7.3,2.1Hz,1H ),8.35(d,J=2.4Hz,1H),8.21-8.13(m,2H),8.03(dd,J=6.6,2.1Hz,1H),7.79( dd,J=9.6,2.8Hz,1H),7.77-7.72(m,1H),7.44(s,1H),6.55(t,J=7.0Hz,1H),4 .02(s,2H),3.50(tt,J=7.6,4.3Hz,1H),1.12-1.05(m,2H),0.99-0.93(m,2H).

[0082] Example 5: Synthesis of compound S5

[0083] Synthesis of intermediate B1 in step 1

[0084] Al (2 g, 16.38 mmol) was dissolved in THF (40 mL), followed by the addition of TEA (4.97 g, 49.14 mmol) and DMAP (0.4 g, 3.28 mmol), then (Boc)₂O (8.22 g, 37.67 mmol). The reaction was allowed to proceed at room temperature for 12 h, and TLC analysis showed complete reaction. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 4.3 g of a white solid, with a yield of 80%. ESI-MS: 323.1 [M+H] + .

[0085] Synthesis of intermediate C2 in step 2

[0086] Fluoroazaindole (0.94 g, 6.21 mmol) and B1 (2 g, 6.21 mmol) were added to isopropanol (20 mL) and water (4 mL), followed by tetramethylguanidine (0.36 g, 3.11 mmol). The reaction was carried out at 25°C for 12 h. TLC analysis showed the reaction was complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 2.5 g of a pale yellow solid, yield 85%. ESI-MS: 458.2 [M+H] + .

[0087] Synthesis of intermediate D2 in step 3

[0088] C2 (2.5 g, 5.28 mmol) was added to a mixed solution of acetonitrile (25 mL) and trifluoroacetic acid (5 mL), followed by triethylsilane (2.46 g, 21.12 mmol). The reaction was carried out at 60°C for 20 h. TLC analysis showed the reaction was complete. After cooling to room temperature, the reaction solution was concentrated. EA (200 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, rotary evaporated, and subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 1.3 g of a pale yellow solid, 95% yield. ESI-MS: 243.1 [M+H] + .

[0089] Step 4 Synthesis of final product S5

[0090] 1-(methyl-D3)-2-oxo-1,2-dihydropyridine-3-carboxylic acid (0.45 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally D2 (0.58 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 0.33 g of a pale yellow solid, with a yield of 36%. ESI-MS: 381.1 [M+H] + .

[0091] 1H NMR (400MHz, DMSO-d6) δ12.45(s,1H),11.72-11.54(m,1H),8.45(dd,J=7.3,2.2Hz,1H),8.34(d,J=2.3Hz,1H),8.2 4-8.12(m,3H),7.76(ddd,J=23.0,9.0,2.6Hz,2H),7.44(d,J=2.5Hz,1H),6.59(dd,J=7.4,6.5Hz,1H),4.01(s,2H).

[0092] Example 6: Synthesis of compound S6

[0093] Synthesis of intermediate B1 in step 1

[0094] Al (2 g, 16.38 mmol) was dissolved in THF (40 mL), followed by the addition of TEA (4.97 g, 49.14 mmol) and DMAP (0.4 g, 3.28 mmol), then (Boc)₂O (8.22 g, 37.67 mmol). The reaction was allowed to proceed at room temperature for 12 h, and TLC analysis showed complete reaction. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 4.3 g of a white solid, with a yield of 80%. ESI-MS: 323.1 [M+H] + .

[0095] Synthesis of intermediate C2 in step 2

[0096] Fluoroazaindole (0.94 g, 6.21 mmol) and B1 (2 g, 6.21 mmol) were added to isopropanol (20 mL) and water (4 mL), followed by tetramethylguanidine (0.36 g, 3.11 mmol). The reaction was carried out at 25°C for 12 h. TLC analysis showed the reaction was complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na₂SO₄, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 2.5 g of a pale yellow solid, yield 85%. ESI-MS: 458.2 [M+H] + .

[0097] Synthesis of intermediate D2 in step 3

[0098] C2 (2.5 g, 5.28 mmol) was added to a mixed solution of acetonitrile (25 mL) and trifluoroacetic acid (5 mL), followed by triethylsilane (2.46 g, 21.12 mmol). The reaction was carried out at 60°C for 20 h. TLC analysis showed the reaction was complete. After cooling to room temperature, the reaction solution was concentrated. EA (200 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, rotary evaporated, and subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 1.3 g of a pale yellow solid, 95% yield. ESI-MS: 243.1 [M+H] + .

[0099] Step 4: Synthesis of final product S6

[0100] 1-Cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid (0.52 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally D2 (0.58 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 0.39 g of a pale yellow solid, with a yield of 40%. ESI-MS: 406.1 [M+H] + .

[0101] 1 H NMR (400MHz, DMSO-d6) δ12.50(s,1H),11.65-11.53(m,1H),8.49-8.44(m,1H),8.34(d,J=2.3Hz,1H),8.29-8.12(m,3H),7.76(ddd ,J=22.4,9.0,2.6Hz,2H),7.44(d,J=2.4Hz,1H),6.66(t,J=7.0Hz,1H),5.22(h,J=6.9Hz,1H),4.02(s,2H),1.38(d,J=6.8Hz,6H).

[0102] Example 7: Synthesis of compound S7

[0103] Synthesis of intermediate B2 in step 1

[0104] In a solution of A2 (25 g, 157.14 mmol) in DMF (0.5 ml) and DCE (250 ml), thionyl chloride (93.48 g, 785.72 mmol) was added dropwise. The mixture was stirred at 70 °C for 4 hours. After the reaction was complete, the mixture was concentrated to obtain a yellow oily product B2 (29.4 g, crude product), which was used directly in the next step.

[0105] Synthesis of intermediate C3 in step 2

[0106] AlCl3 (54.27 g, 407 mmol) was added to a DCM (230 mL) solution of 5-chloro-1H-pyrrolo[2,3-b]pyridine (23 g, 150.74 mmol) at 0 °C for 0.5 h. Then, a solution of crude B2 (29.44 g, 165.81 mmol) in DCM (400 mL) was added. The reaction mixture was stirred at 25 °C for 16 h, and then quenched with saturated brine. The pH was adjusted to 7-8 using NaHCO3. The mixture was filtered and concentrated under reduced pressure to obtain a residue. Water (600 mL) was added, and the residue was extracted with EA (600 mL × 2). The combined organic phases were washed with brine (500 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The concentrated product was ground with EA to obtain C3 (44 g, crude) as a grayish-white solid, which was used directly in the next step.

[0107] Synthesis of intermediate D3 in step 3

[0108] C3 (40 g, 136.21 mmol) was added to DMSO (400 mL) solution with TEA (41.35 g, 408.64 mmol), followed by dropwise addition of 4-methoxybenzylamine (14.95 g, 108.97 mmol). The mixture was stirred at 25 °C for 16 h. Water (550 mL) was added and the mixture was extracted with EA (2000 mL × 2). The combined organic phases were washed with brine (2000 mL), dried over anhydrous sodium sulfate, and concentrated. The concentrate was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give a crude product, which was purified by preparative HPLC (water (0.1% TFA)-acetonitrile) to give 10 g of a pale white solid, yield 18%. ESI-MS: 411.2 [M+H] + .

[0109] Synthesis of intermediate E1 in step 4

[0110] D3 (10 g, 24.34 mmol) was dissolved in THF (100 mL), and LiAlH4 (14.6 mL, 36.51 mmol, 2.5 M in THF) solution was slowly added dropwise at 0 °C. The mixture was stirred at 25 °C for 5 h under a nitrogen atmosphere. The reaction mixture was poured into ice water and extracted with EA (100 mL × 2). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 2.50 g of white solid, yield 25%. ESI-MS: 413.2 [M + H] + .

[0111] Synthesis of intermediate F1 in step 5

[0112] Et3SiH (10.42 g, 89.62 mmol) and TFA (18.39 g, 161.32 mmol) were added to E1 (7.40 g, 17.92 mmol) in DCM (80 mL) solution at 0 °C. The mixture was stirred at 25 °C for 1 h. After the reaction was complete, the mixture was evaporated to dryness and saturated brine was added. The pH was adjusted to 7-8 with NaHCO3. The filter cake was collected by vacuum filtration, washed with 100 mL of water, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 2.50 g of a light white solid, yield 35%. ESI-MS: 397.2 [M+H] + .

[0113] Step 6 Synthesis of intermediate G1

[0114] TfOH (1.42 g, 9.45 mmol) was added to F1 (2.50 g, 6.30 mmol) in DCM (50 mL) solution at 0 °C. The mixture was stirred at 25 °C for 2 h, and the reaction was monitored by TLC until complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water, then washing with saturated NaCl, drying over anhydrous sodium sulfate, filtering, rotary evaporation, and silica gel column chromatography (dichloromethane:tetrahydrofuran = 1:1) to give 1.1 g of a grayish-white solid, yield 65%. ESI-MS: 277.1 [M+H] + .

[0115] Step 7: Synthesis of final product S7

[0116] 2-O-1-methylnicotinic acid (0.5 g, 2.86 mmol) was dissolved in DMF (5 ml), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally G1 (0.66 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 ml) was added, followed by washing with 50 ml of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 0.33 g of a pale white solid, yield 34%. ESI-MS: 412.1 [M+H] + .

[0117] 1 H NMR (400MHz, DMSO-d6) δ12.51(s,1H),11.75(s,1H),8.46(dd,J=7.4,2.2Hz,1H),8.19(dt,J=4.9,2.3Hz,2H),8.11 -8.07(m,1H),8.03(d,J=2.4Hz,1H),7.91-7.83(m,1H),7.40(s,1H),6.64-6.57(m,1H),4.02(s,2H),3.63(s,3H).

[0118] Example 8: Synthesis of compound S8

[0119] Synthesis of intermediate B2 in step 1

[0120] In a solution of A2 (25 g, 157.14 mmol) in DMF (0.5 ml) and DCE (250 ml), thionyl chloride (93.48 g, 785.72 mmol) was added dropwise. The mixture was stirred at 70 °C for 4 hours. After the reaction was complete, the mixture was concentrated to obtain a yellow oily product B2 (29.4 g, crude product), which was used directly in the next step.

[0121] Synthesis of intermediate C3 in step 2

[0122] AlCl3 (54.27 g, 407 mmol) was added to a DCM (230 mL) solution of 5-chloro-1H-pyrrolo[2,3-b]pyridine (23 g, 150.74 mmol) at 0 °C for 0.5 h. Then, a solution of crude B2 (29.44 g, 165.81 mmol) in DCM (400 mL) was added. The reaction mixture was stirred at 25 °C for 16 h, and then quenched with saturated brine. The pH was adjusted to 7-8 using NaHCO3. The mixture was filtered and concentrated under reduced pressure to obtain a residue. Water (600 mL) was added, and the residue was extracted with EA (600 mL × 2). The combined organic phases were washed with brine (500 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The concentrated product was ground with EA to obtain C3 (44 g, crude) as a grayish-white solid, which was used directly in the next step.

[0123] Synthesis of intermediate D3 in step 3

[0124] C3 (40 g, 136.21 mmol) was added to DMSO (400 mL) solution with TEA (41.35 g, 408.64 mmol), followed by dropwise addition of 4-methoxybenzylamine (14.95 g, 108.97 mmol). The mixture was stirred at 25 °C for 16 h. Water (550 mL) was added and the mixture was extracted with EA (2000 mL × 2). The combined organic phases were washed with brine (2000 mL), dried over anhydrous sodium sulfate, and concentrated. The concentrate was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give a crude product, which was purified by preparative HPLC (water (0.1% TFA)-acetonitrile) to give 10 g of a pale white solid, yield 18%. ESI-MS: 411.2 [M+H] + .

[0125] Synthesis of intermediate E1 in step 4

[0126] D3 (10 g, 24.34 mmol) was dissolved in THF (100 mL), and LiAlH4 (14.6 mL, 36.51 mmol, 2.5 M in THF) solution was slowly added dropwise at 0 °C. The mixture was stirred at 25 °C for 5 h under a nitrogen atmosphere. The reaction mixture was poured into ice water and extracted with EA (100 mL × 2). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 2.50 g of white solid, yield 25%. ESI-MS: 413.2 [M + H] + .

[0127] Synthesis of intermediate F1 in step 5

[0128] Et3SiH (10.42 g, 89.62 mmol) and TFA (18.39 g, 161.32 mmol) were added to E1 (7.40 g, 17.92 mmol) in DCM (80 mL) solution at 0 °C. The mixture was stirred at 25 °C for 1 h. After the reaction was complete, the mixture was evaporated to dryness and saturated brine was added. The pH was adjusted to 7-8 with NaHCO3. The filter cake was collected by vacuum filtration, washed with 100 mL of water, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 2.50 g of a light white solid, yield 35%. ESI-MS: 397.2 [M+H] + .

[0129] Step 6 Synthesis of intermediate G1

[0130] TfOH (1.42 g, 9.45 mmol) was added to F1 (2.50 g, 6.30 mmol) in DCM (50 mL) solution at 0 °C. The mixture was stirred at 25 °C for 2 h, and the reaction was monitored by TLC until complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water, then washing with saturated NaCl, drying over anhydrous sodium sulfate, filtering, rotary evaporation, and silica gel column chromatography (dichloromethane:tetrahydrofuran = 1:1) to give 1.1 g of a grayish-white solid, yield 65%. ESI-MS: 277.1 [M+H] + .

[0131] Step 7 Synthesis of final product S8

[0132] 1-Cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid (0.43 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally G1 (0.66 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to give 0.37 g of a pale white solid, with a yield of 36%. ESI-MS: 438.1 [M+H] + .

[0133] 1H NMR (400MHz, DMSO-d6) δ12.51(s,1H),11.75(s,1H),8.44(dd,J=7.4,2.1Hz,1H),8.19(d,J=2.3Hz,1H),8.06(ddd,J =15.6,9.1,2.0Hz,3H),7.91-7.84(m,1H),7.41(d,J=2.3Hz,1H),4.03(s,2H),1.13-1.05(m,2H),1.00-0.92(m,2H).

[0134] Example 9: Synthesis of compound S9

[0135] Synthesis of intermediate B2 in step 1

[0136] In a solution of A2 (25 g, 157.14 mmol) in DMF (0.5 ml) and DCE (250 ml), thionyl chloride (93.48 g, 785.72 mmol) was added dropwise. The mixture was stirred at 70 °C for 4 hours. After the reaction was complete, the mixture was concentrated to obtain a yellow oily product B2 (29.4 g, crude product), which was used directly in the next step.

[0137] Synthesis of intermediate C3 in step 2

[0138] AlCl3 (54.27 g, 407 mmol) was added to a DCM (230 mL) solution of 5-chloro-1H-pyrrolo[2,3-b]pyridine (23 g, 150.74 mmol) at 0 °C for 0.5 h. Then, a solution of crude B2 (29.44 g, 165.81 mmol) in DCM (400 mL) was added. The reaction mixture was stirred at 25 °C for 16 h, and then quenched with saturated brine. The pH was adjusted to 7-8 using NaHCO3. The mixture was filtered and concentrated under reduced pressure to obtain a residue. Water (600 mL) was added, and the residue was extracted with EA (600 mL × 2). The combined organic phases were washed with brine (500 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The concentrated product was ground with EA to obtain C3 (44 g, crude) as a grayish-white solid, which was used directly in the next step.

[0139] Synthesis of intermediate D3 in step 3

[0140] C3 (40 g, 136.21 mmol) was added to DMSO (400 mL) solution with TEA (41.35 g, 408.64 mmol), followed by dropwise addition of 4-methoxybenzylamine (14.95 g, 108.97 mmol). The mixture was stirred at 25 °C for 16 h. Water (550 mL) was added and the mixture was extracted with EA (2000 mL × 2). The combined organic phases were washed with brine (2000 mL), dried over anhydrous sodium sulfate, and concentrated. The concentrate was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give a crude product, which was purified by preparative HPLC (water (0.1% TFA)-acetonitrile) to give 10 g of a pale white solid, yield 18%. ESI-MS: 411.2 [M+H] + .

[0141] Synthesis of intermediate E1 in step 4

[0142] D3 (10 g, 24.34 mmol) was dissolved in THF (100 mL), and LiAlH4 (14.6 mL, 36.51 mmol, 2.5 M in THF) solution was slowly added dropwise at 0 °C. The mixture was stirred at 25 °C for 5 h under a nitrogen atmosphere. The reaction mixture was poured into ice water and extracted with EA (100 mL × 2). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 2.50 g of white solid, yield 25%. ESI-MS: 413.2 [M + H] + .

[0143] Synthesis of intermediate F1 in step 5

[0144] Et3SiH (10.42 g, 89.62 mmol) and TFA (18.39 g, 161.32 mmol) were added to E1 (7.40 g, 17.92 mmol) in DCM (80 mL) solution at 0 °C. The mixture was stirred at 25 °C for 1 h. After the reaction was complete, the mixture was evaporated to dryness and saturated brine was added. The pH was adjusted to 7-8 with NaHCO3. The filter cake was collected by vacuum filtration, washed with 100 mL of water, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 2.50 g of a light white solid, yield 35%. ESI-MS: 397.2 [M+H] + .

[0145] Step 6 Synthesis of intermediate G1

[0146] TfOH (1.42 g, 9.45 mmol) was added to F1 (2.50 g, 6.30 mmol) in DCM (50 mL) solution at 0 °C. The mixture was stirred at 25 °C for 2 h, and the reaction was monitored by TLC until complete. 100 mL of ethyl acetate was added, followed by washing with 50 mL of water, then washing with saturated NaCl, drying over anhydrous sodium sulfate, filtering, rotary evaporation, and silica gel column chromatography (dichloromethane:tetrahydrofuran = 1:1) to give 1.1 g of a grayish-white solid, yield 65%. ESI-MS: 277.1 [M+H] + .

[0147] Step 7 Synthesis of final product S9

[0148] Under a nitrogen atmosphere at room temperature, phenyl chloroformate (105 mg, 0.68 mmol) and DIEA (115 mg, 0.90 mmol) were added to a solution of G1 (127 mg, 0.45 mmol) in 3 mL of THF. The reaction mixture was stirred at room temperature for 1 h. The mixture was diluted with water (10 mL) and extracted with EA (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Under a nitrogen atmosphere at room temperature, pyridine (3 mL) and 3,3-dimethylpyrrole hydrochloride (90 mg, 0.90 mmol) were added to the concentrate. The reaction mixture was stirred at 40 °C for 1 h. The mixture was diluted with water (20 mL) and extracted with EA (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 120 mg of white solid in 67% yield. ESI-MS: 402.1 [M+H] + .

[0149] 1 H NMR (400MHz, DMSO-d6) δ10.24(d,J=6.6Hz,1H),9.78(s,1H),8.25(d,J=1.8Hz,1H),7.82(d,J=2.0Hz,1H),7.62(dd,J=8.8,5.1Hz,1H),7 .46(d,J=8.8Hz,1H),7.02(d,J=6.8Hz,1H),3.87(s,2H),3.51-3.34(m,2H),1.81(dd,J=5.6,3.8Hz,1H),1.72-1.65(m,1H),0.98(s,5H).

[0150] Example 10: Synthesis of compound S10

[0151] Synthesis of intermediate B3 in step 1

[0152] Under a nitrogen atmosphere at 0 °C, NaH (60%, 2.0 g, 51.0 mmol) was added to a 50 mL solution of A3 (5.0 g, 25.5 mmol) in THF. The reaction was stirred at 0 °C for 0.5 h. Then, SEMCl (6.4 g, 38.3 mmol) was added, and the reaction was stirred at room temperature for 2 h. The reaction was confirmed to be complete by TLC. The mixture was diluted with an aqueous solution of ammonium chloride (100 mL) and extracted with EA (50 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was achieved by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give 7.0 g of a yellow oil, in 84% yield. ESI-MS: 327.9 [M+H] + .

[0153] Synthesis of intermediate C4 in step 2

[0154] Under a nitrogen atmosphere at room temperature, 1-methyl-4-pyrazolone boronic acid tinane ester (1.4 g, 6.8 mmol), K₂CO₃ (2 M, 9.2 mL, 18.4 mmol), and Pd(dppf)Cl₂ (498 mg, 0.68 mmol) were added to a solution of B₃ (2.0 g, 6.1 mmol) dioxane (36.8 mL). The reaction was stirred at 85 °C for 5 h. The reaction was monitored by TLC until complete. The mixture was diluted with water (50 mL) and extracted with EA (30 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 2.0 g of a yellow oil, 99% yield. ESI-MS: 329.5 [M + H] + .

[0155] Synthesis of intermediate D4 in step 3

[0156] Under a nitrogen atmosphere at 0°C, NBS (814 mg, 4.6 mmol) and AIBN (150 mg, 0.91 mmol) were added to a CCl4 (1.5 g, 4.6 mmol) solution in 20 mL of CCl4. The reaction mixture was stirred at 40°C for 2 h. After the reaction was complete, the mixture was diluted with water (30 mL) and extracted with DCM (20 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give 1.6 g of a yellow solid, yield 86%. ESI-MS: 408.7 [M+H] + .

[0157] Synthesis of intermediate E2 in step 4

[0158] Under a nitrogen atmosphere at room temperature, dioxane (990 mg, 3.7 mmol), KOH (207 mg, 3.7 mmol), and Pd[P(t-Bu)3]2 (97 mg, 0.19 mmol) were added to a solution of D4 (750 mg, 1.9 mmol) in dioxane (36 mL) and H2O (3 mL). The reaction mixture was stirred at 80 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure by TLC. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) yielded 860 mg of a yellow oil (99% yield). ESI-MS: 469.5 [M+H] + .

[0159] Synthesis of intermediate F2 in step 5

[0160] Under a nitrogen atmosphere at room temperature, 2-amino-5-bromo-6-methylpyridine (687 mg, 3.7 mmol), KOH (206 mg, 3.7 mmol), and Pd[P(t-Bu)3]2 (92 mg, 0.18 mmol) were added to a solution of E2 (860 mg, 1.8 mmol) in dioxane (36 mL) and water (3 mL). The reaction mixture was stirred at 80 °C for 1 h. After the reaction was complete, the mixture was diluted with water (50 mL) and extracted with EA (30 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl oleate = 2:1) to give 200 mg of a yellow solid, yield 24%. ESI-MS: 449.5 [M+H] + .

[0161] Synthesis of intermediate G2 in step 6

[0162] Under a nitrogen atmosphere at room temperature, phenyl chloroformate (105 mg, 0.68 mmol) and DIEA (115 mg, 0.90 mmol) were added to a solution of F2 (200 mg, 0.45 mmol) in 3 mL of THF. The reaction mixture was stirred at room temperature for 1 h. The mixture was diluted with water (10 mL) and extracted with EA (10 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Under a nitrogen atmosphere at room temperature, pyridine (3 mL) and 6-azaspiro[3,4]octane (100 mg, 0.90 mmol) were added to the concentrate. The reaction mixture was stirred at 40 °C for 1 h. The mixture was diluted with water (20 mL) and extracted with EA (10 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give 174 mg of a yellow solid, yield 67%. ESI-MS: 587.3 [M+H] + .

[0163] Step 7 Synthesis of final product S10

[0164] At 0 °C, TFA (1 mL) and Et3SiH (0.2 mL) were added to a 2 mL solution of G2 (174 mg, 0.30 mmol) in DCM. The reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. ACN (1.5 mL) and NH3 / H2O (0.5 mL) were added to the concentrate at room temperature. The reaction mixture was stirred at room temperature for 16 h. The mixture was concentrated under reduced pressure. The mixture was purified by preparative HPLC (0.1% NH4HCO3, 60% ACN in H2O solution) to give 52.5 mg of white solid, yield 39%. ESI-MS: 456.1 [M+H] + .

[0165] 1 H NMR (400MHz, Chloroform-d) δ9.23 (s, 1H), 8.44 (d, J = 2.0Hz, 1H), 7.87 (d, J = 8. 4Hz,1H),7.80(dd,J=2.0,0.4Hz,1H),7.75(d,J=0.8Hz,1H),7.61(d,J=0.8Hz, 1H),7.43(d,J=8.4Hz,1H),7.01(s,1H),6.88(d,J=2.4Hz,1H),4.00(s,2H),3. 96(s,3H),3.48(t,J=6.8Hz,2H),3.43(s,2H),2.42(s,3H),2.06-1.87(m,8H).

[0166] Example 11: Synthesis of compound S11

[0167] Synthesis of intermediate B3 in step 1

[0168] Under a nitrogen atmosphere at 0 °C, NaH (60%, 2.0 g, 51.0 mmol) was added to a 50 mL solution of A3 (5.0 g, 25.5 mmol) in THF. The reaction was stirred at 0 °C for 0.5 h. Then, SEMCl (6.4 g, 38.3 mmol) was added, and the reaction was stirred at room temperature for 2 h. The reaction was confirmed to be complete by TLC. The mixture was diluted with an aqueous solution of ammonium chloride (100 mL) and extracted with EA (50 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was achieved by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give 7.0 g of a yellow oil, in 84% yield. ESI-MS: 327.9 [M+H] + .

[0169] Synthesis of intermediate C4 in step 2

[0170] Under a nitrogen atmosphere at room temperature, 1-methyl-4-pyrazolone boronic acid tinane ester (1.4 g, 6.8 mmol), K₂CO₃ (2 M, 9.2 mL, 18.4 mmol), and Pd(dppf)Cl₂ (498 mg, 0.68 mmol) were added to a solution of B₃ (2.0 g, 6.1 mmol) dioxane (36.8 mL). The reaction was stirred at 85 °C for 5 h. The reaction was monitored by TLC until complete. The mixture was diluted with water (50 mL) and extracted with EA (30 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 2.0 g of a yellow oil, 99% yield. ESI-MS: 329.5 [M + H] + .

[0171] Synthesis of intermediate D4 in step 3

[0172] Under a nitrogen atmosphere at 0°C, NBS (814 mg, 4.6 mmol) and AIBN (150 mg, 0.91 mmol) were added to a CCl4 (1.5 g, 4.6 mmol) solution in 20 mL of CCl4. The reaction mixture was stirred at 40°C for 2 h. After the reaction was complete, the mixture was diluted with water (30 mL) and extracted with DCM (20 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give 1.6 g of a yellow solid, yield 86%. ESI-MS: 408.7 [M+H] + .

[0173] Synthesis of intermediate E2 in step 4

[0174] Under a nitrogen atmosphere at room temperature, dioxane (990 mg, 3.7 mmol), KOH (207 mg, 3.7 mmol), and Pd[P(t-Bu)3]2 (97 mg, 0.19 mmol) were added to a solution of D4 (750 mg, 1.9 mmol) in dioxane (36 mL) and H2O (3 mL). The reaction mixture was stirred at 80 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure by TLC. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) yielded 860 mg of a yellow oil (99% yield). ESI-MS: 469.5 [M+H] + .

[0175] Synthesis of intermediate F2 in step 5

[0176] Under a nitrogen atmosphere at room temperature, 2-amino-5-bromo-6-methylpyridine (687 mg, 3.7 mmol), KOH (206 mg, 3.7 mmol), and Pd[P(t-Bu)3]2 (92 mg, 0.18 mmol) were added to a solution of E2 (860 mg, 1.8 mmol) in dioxane (36 mL) and water (3 mL). The reaction mixture was stirred at 80 °C for 1 h. After the reaction was complete, the mixture was diluted with water (50 mL) and extracted with EA (30 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl oleate = 2:1) to give 200 mg of a yellow solid, yield 24%. ESI-MS: 449.5 [M+H] + .

[0177] Step 6 Synthesis of intermediate G3

[0178] At 0 °C, TFA (1 mL) and Et3SiH (0.2 mL) were added to a 2 mL solution of F2 (134 mg, 0.30 mmol) in DCM. The reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. ACN (1.5 mL) and NH3 / H2O (0.5 mL) were added to the concentrate at room temperature. The reaction mixture was stirred at room temperature for 16 h. The mixture was concentrated under reduced pressure. The solution was purified by silica gel column chromatography (dichloromethane:methanol = 15:1) to give 38 mg of a white solid, 40% yield. ESI-MS: 319.1 [M+H] + .

[0179] Step 7 Synthesis of final product S11

[0180] 2-O-1-methylnicotinic acid (0.5 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally G3 (0.76 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, and the mixture was washed with 50 mL of water and saturated NaCl. The solution was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 0.58 g of a pale yellow solid, yield 54%. ESI-MS: 454.1 [M+H] + .

[0181] 1H NMR (400MHz, DMSO-d6) δ12.37(s,1H),11.45-11.35(m,2H),8.45(td,J=3.8,2.2Hz,2H),8.16(dd,J=6.5,2.2Hz,2H),8.12(s,1H),8. 04-7.99(m,2H),7.85(s,1H),7.59(d,J=8.4Hz,1H),7.15(d,J=2.4Hz,1H),6.62-6.55(m,2H),4.04(s,2H),3.86(s,3H),3.62(s,3H).

[0182] Example 12: Synthesis of compound S12

[0183] Synthesis of intermediate B3 in step 1

[0184] Under a nitrogen atmosphere at 0 °C, NaH (60%, 2.0 g, 51.0 mmol) was added to a 50 mL solution of A3 (5.0 g, 25.5 mmol) in THF. The reaction was stirred at 0 °C for 0.5 h. Then, SEMCl (6.4 g, 38.3 mmol) was added, and the reaction was stirred at room temperature for 2 h. The reaction was confirmed to be complete by TLC. The mixture was diluted with an aqueous solution of ammonium chloride (100 mL) and extracted with EA (50 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was achieved by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give 7.0 g of a yellow oil, in 84% yield. ESI-MS: 327.9 [M+H] + .

[0185] Synthesis of intermediate C4 in step 2

[0186] Under a nitrogen atmosphere at room temperature, 1-methyl-4-pyrazolone boronic acid tinane ester (1.4 g, 6.8 mmol), K₂CO₃ (2 M, 9.2 mL, 18.4 mmol), and Pd(dppf)Cl₂ (498 mg, 0.68 mmol) were added to a solution of B₃ (2.0 g, 6.1 mmol) dioxane (36.8 mL). The reaction was stirred at 85 °C for 5 h. The reaction was monitored by TLC until complete. The mixture was diluted with water (50 mL) and extracted with EA (30 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 2.0 g of a yellow oil, 99% yield. ESI-MS: 329.5 [M + H] + .

[0187] Synthesis of intermediate D4 in step 3

[0188] Under a nitrogen atmosphere at 0°C, NBS (814 mg, 4.6 mmol) and AIBN (150 mg, 0.91 mmol) were added to a CCl4 (1.5 g, 4.6 mmol) solution in 20 mL of CCl4. The reaction mixture was stirred at 40°C for 2 h. After the reaction was complete, the mixture was diluted with water (30 mL) and extracted with DCM (20 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give 1.6 g of a yellow solid, yield 86%. ESI-MS: 408.7 [M+H] + .

[0189] Synthesis of intermediate E2 in step 4

[0190] Under a nitrogen atmosphere at room temperature, dioxane (990 mg, 3.7 mmol), KOH (207 mg, 3.7 mmol), and Pd[P(t-Bu)3]2 (97 mg, 0.19 mmol) were added to a solution of D4 (750 mg, 1.9 mmol) in dioxane (36 mL) and H2O (3 mL). The reaction mixture was stirred at 80 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure by TLC. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) yielded 860 mg of a yellow oil (99% yield). ESI-MS: 469.5 [M+H] + .

[0191] Synthesis of intermediate F2 in step 5

[0192] Under a nitrogen atmosphere at room temperature, 2-amino-5-bromo-6-methylpyridine (687 mg, 3.7 mmol), KOH (206 mg, 3.7 mmol), and Pd[P(t-Bu)3]2 (92 mg, 0.18 mmol) were added to a solution of E2 (860 mg, 1.8 mmol) in dioxane (36 mL) and water (3 mL). The reaction mixture was stirred at 80 °C for 1 h. After the reaction was complete, the mixture was diluted with water (50 mL) and extracted with EA (30 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl oleate = 2:1) to give 200 mg of a yellow solid, yield 24%. ESI-MS: 449.5 [M+H] + .

[0193] Step 6 Synthesis of intermediate G3

[0194] At 0 °C, TFA (1 mL) and Et3SiH (0.2 mL) were added to a 2 mL solution of F2 (134 mg, 0.30 mmol) in DCM. The reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. ACN (1.5 mL) and NH3 / H2O (0.5 mL) were added to the concentrate at room temperature. The reaction mixture was stirred at room temperature for 16 h. The mixture was concentrated under reduced pressure. The solution was purified by silica gel column chromatography (dichloromethane:methanol = 15:1) to give 38 mg of a white solid, 40% yield. ESI-MS: 319.1 [M+H] + .

[0195] Step 7: Synthesis of final product S12

[0196] 2-Hydroxy-3-pyridinecarboxylic acid (0.4 g, 2.86 mmol) was dissolved in DMF (5 mL), followed by DIPEA (0.92 g, 7.14 mmol), then HATU (2.71 g, 2.97 mmol). The mixture was stirred for 2 min, and finally G3 (0.58 g, 2.38 mmol) was added. The reaction was allowed to proceed at room temperature for 24 h. TLC analysis showed the reaction was complete. EA (100 mL) was added, followed by washing with 50 mL of water and saturated NaCl. The mixture was dried over anhydrous Na2SO4, filtered, and rotary evaporated. The crude product was then subjected to silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain 0.32 g of a pale yellow solid, with a yield of 30%. ESI-MS: 440.1 [M+H] + .

[0197] 1 H NMR (400MHz, DMSO-d6) δ12.12(d,J=6.6Hz,1H),11.73(s,1H),10.39(d,J=6 .6Hz,1H),8.59(d,J=1.6Hz,1H),8.35-8.28(m,2H),8.16(s,1H),7.99(s,1 H),7.90(td,J=6.9,1.3Hz,1H),7.38(d,J=7.9Hz,1H),7.32(d,J=7.9Hz,1H ), 7.00 (d, J = 6.5Hz, 1H), 6.70 (s, 1H), 6.70 (d, J = 14.3Hz, 1H), 3.97 (s, 2H).

[0198] Example 13: Synthesis of compound S13

[0199] Synthesis of intermediate B4 in step 1

[0200] NaH (2.23 g, 55.9 mmol) was added to A4 (15.0 g, 46.6 mmol) in DMF (200 mL) solution and stirred at 0 °C for 0.5 h. Then, benzenesulfonyl chloride (9.8 g, 55.9 mmol) was added to the above solution at 0 °C, and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was poured into water (800 mL), and a large amount of solid precipitate precipitated. The solid was collected by filtration, washed with water (100 mL × 3), and dried under high vacuum to give 17.7 g of white compound, yield 82%. ESI-MS: 463.0 [M+H] + .

[0201] Synthesis of intermediate D5 in step 2

[0202] To a solution of C5 (4.5 g, 29.0 mmol) in dioxane (50 mL), BocNH2 (8.4 g, 72.6 mmol), Pd2(dba)3 (1.3 g, 1.45 mmol), X-phos (1.3 g, 2.90 mmol), and Cs2CO3 (2.6 g, 8.06 mmol) were added. The reaction mixture was heated to 90 °C and stirred for 16 h under a nitrogen atmosphere. The reaction mixture was quenched with water (30 mL). The mixture was extracted with EA (50 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 25:1) to give 3.0 g of a yellow solid, 44% yield. ESI-MS: 237.1 [M+H] + .

[0203] Synthesis of intermediate E3 in step 3

[0204] At 0 °C, iPrMgCl (2.0 M, 9.5 mL, 19.1 mmol) was added dropwise to a THF (100 mL) solution of B4 (3, 8.7 g, 19.1 mmol) and stirred at room temperature under a nitrogen atmosphere for 1 h. D5 (1.5 g, 6.4 mmol) was added to the above solution at 0 °C and stirred again at room temperature under a nitrogen atmosphere for 1 h. The reaction mixture was quenched with water (30 mL). The mixture was extracted with EA (50 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained residues were purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 1.9 g of white solid, 52% yield. ESI-MS: 573.2 [M + H] + .

[0205] Synthesis of intermediate F3 in step 4

[0206] TFA (2 mL) and Et3SiH (3 mL) were added to a DCM (30 mL) solution of E3 (4 g, 1.9 g, 3.32 mmol), and the mixture was stirred at 40 °C for 5 h under a nitrogen atmosphere. The mixture was concentrated under reduced pressure. The pH was adjusted to 8 with NH4OH. The mixture was extracted with DCM (30 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was achieved by silica gel column chromatography (dichloromethane:methanol = 99:1) to give 710 mg of a yellow solid, 47% yield. ESI-MS: 457.1 [M+H] + .

[0207] Step 5 Synthesis of intermediate G4

[0208] Morpholine (68.9 mg, 1.03 mmol), Pd₂(dba)₃ (78.3 mg, 0.09 mmol), Ru-phos (79.8 mg, 0.17 mmol), and Cs₂CO₃ (836.0 mg, 0.20 mmol) were added to a solution of F₃ (390 mg, 0.86 mmol) in dioxane (20 mL). The reaction mixture was heated to 110 °C and stirred under a nitrogen atmosphere for 6 h. The reaction mixture was quenched with water (20 mL). The mixture was extracted with EA (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (dichloromethane:methanol = 99:1) to give 225 mg of a yellow solid, 57% yield. ESI-MS: 464.2 [M+H] + .

[0209] Synthesis of intermediate H1 in step 6

[0210] To a solution of G4 (200 mg, 0.43 mmol) in THF (4 mL), phenyl chloroformate (101 mg, 0.65 mmol) and DIEA (110 mg, 0.86 mmol) were added, and the mixture was stirred for 1 h at room temperature under a nitrogen atmosphere. The reaction mixture was quenched with water (10 mL). The mixture was extracted with EA (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was dissolved in pyridine (6 mL), and then 6-aza[3,4]octane (95.9 mg, 0.86 mmol) was added, and the mixture was stirred for 1 h at 40 °C under a nitrogen atmosphere. The reaction was quenched with water (20 mL). The mixture was extracted with EA (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (dichloromethane:methanol = 99:1) to give 100 mg of a yellow solid, 39% yield. ESI-MS: 601.2 [M+H] + .

[0211] Step 7 Synthesis of final product S13

[0212] Add nBu4NOH (434 mg, 0.67 mmol, 40 wt% in H2O) to a solution of H1 (100 mg, 0.17 mmol) in 5 mL of THF, and stir for 16 h at 50 °C under a nitrogen atmosphere. Quench the reaction mixture with water (15 mL). Extract the mixture with EA (20 mL x 3). Wash the combined organic layers with brine, dry with anhydrous sodium sulfate, filter, and concentrate. Purify by Prep-HPLC (mobile phase: [H2O (0.04% NH4HCO3)-ACN]) to give 54.7 mg of white solid, yield 71%. ESI-MS: 461.2 [M+H] +

[0213] 1 H NMR (400MHz, DMSO-d6) δ8.14 (s, 1H), 8.07-8.04 (m, 2H), 7.39 (d, J = 8.8Hz, 1H), 7.29 (s, 1H), 4.20 (s,2H),3.90-3.87(m,4H),3.58-3.43(m,4H),3.25-3.23(m,4H),2.68(s,3H),2.08-1.93(m,8H).

[0214] Example 14: Synthesis of compound S14

[0215] Synthesis of intermediate B4 in step 1

[0216] NaH (2.23 g, 55.9 mmol) was added to A4 (15.0 g, 46.6 mmol) in DMF (200 mL) solution and stirred at 0 °C for 0.5 h. Then, benzenesulfonyl chloride (9.8 g, 55.9 mmol) was added to the above solution at 0 °C, and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was poured into water (800 mL), and a large amount of solid precipitate precipitated. The solid was collected by filtration, washed with water (100 mL × 3), and dried under high vacuum to give 17.7 g of white compound, yield 82%. ESI-MS: 463.0 [M+H] + .

[0217] Synthesis of intermediate D5 in step 2

[0218] To a solution of C5 (4.5 g, 29.0 mmol) in dioxane (50 mL), BocNH2 (8.4 g, 72.6 mmol), Pd2(dba)3 (1.3 g, 1.45 mmol), X-phos (1.3 g, 2.90 mmol), and Cs2CO3 (2.6 g, 8.06 mmol) were added. The reaction mixture was heated to 90 °C and stirred for 16 h under a nitrogen atmosphere. The reaction mixture was quenched with water (30 mL). The mixture was extracted with EA (50 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 25:1) to give 3.0 g of a yellow solid, 44% yield. ESI-MS: 237.1 [M+H] + .

[0219] Synthesis of intermediate E3 in step 3

[0220] At 0 °C, iPrMgCl (2.0 M, 9.5 mL, 19.1 mmol) was added dropwise to a solution of B4 (3, 8.7 g, 19.1 mmol) in 100 mL of THF, and the mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. D5 (1.5 g, 6.4 mmol) was added to the above solution at 0 °C, and the mixture was stirred again at room temperature under a nitrogen atmosphere for 1 h. The reaction mixture was quenched with water (30 mL). The mixture was extracted with EA (50 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained residues were purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 1.9 g of a white solid, 52% yield. ESI-MS: 573.2 [M + H] +

[0221] Synthesis of intermediate F3 in step 4

[0222] TFA (2 mL) and Et3SiH (3 mL) were added to a DCM (30 mL) solution of E3 (4 g, 1.9 g, 3.32 mmol), and the mixture was stirred at 40 °C for 5 h under a nitrogen atmosphere. The mixture was concentrated under reduced pressure. The pH was adjusted to 8 with NH4OH. The mixture was extracted with DCM (30 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was achieved by silica gel column chromatography (dichloromethane:methanol = 99:1) to give 710 mg of a yellow solid, 47% yield. ESI-MS: 457.1 [M+H] + .

[0223] Synthesis of intermediate G5 in step 5

[0224] To a solution of F3 (390 mg, 0.86 mmol) in dioxane (20 mL), N-methylpiperazine (103 mg, 1.03 mmol), Pd2(dba)3 (78.3 mg, 0.09 mmol), Ru-phos (79.8 mg, 0.17 mmol), and Cs2CO3 (836.0 mg, 0.20 mmol) were added. The reaction mixture was heated to 110 °C and stirred under a nitrogen atmosphere for 6 h. The reaction mixture was quenched with water (20 mL). The mixture was extracted with EA (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (dichloromethane:methanol = 99:1) gave 217 mg of a yellow solid, 53% yield. ESI-MS: 477.2 [M+H] + .

[0225] Synthesis of intermediate H2 in step 6

[0226] To a solution of G5 (205 mg, 0.43 mmol) in THF (4 mL), phenyl chloroformate (101 mg, 0.65 mmol) and DIEA (110 mg, 0.86 mmol) were added, and the mixture was stirred for 1 h at room temperature under a nitrogen atmosphere. The reaction mixture was quenched with water (10 mL). The mixture was extracted with EA (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was dissolved in pyridine (6 mL), and then 6-aza[3,4]octane (95.9 mg, 0.86 mmol) was added, and the mixture was stirred for 1 h at 40 °C under a nitrogen atmosphere. The reaction was quenched with water (20 mL). The mixture was extracted with EA (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (dichloromethane:methanol = 99:1) to give 95 mg of a yellow solid, 36% yield. ESI-MS: 614.3 [M+H] + .

[0227] Step 7: Synthesis of final product S14

[0228] Add nBu4NOH (434 mg, 0.67 mmol, 40 wt% in H2O) to a solution of H1 (104 mg, 0.17 mmol) in 5 mL of THF, and stir for 16 h at 50 °C under a nitrogen atmosphere. Quench the reaction mixture with water (15 mL). Extract the mixture with EA (20 mL × 3). Wash the combined organic layers with brine, dry with anhydrous sodium sulfate, filter, and concentrate. Purify by Prep-HPLC (mobile phase: [H2O (0.04% NH4HCO3)-ACN]) to give 28.8 mg of white solid, yield 35%. ESI-MS: 474.0 [M+H] +

[0229] 1 H NMR (400MHz, CD3OD-d6)7.62(d,J=8.4Hz,1H),7.48(d,J=4.9Hz,1H),7.47(s,1H),7.01(s,1H),4.01(s,2H),3.45 (t,J=6.8Hz,2H),3.41(s,2H),3.27-3.22(m,4H),3.16-3.11(m,4H),2.72(s,3H),2.40(s,3H),2.05-1.91(m,8H).

[0230] Example 15 Enzymatic activity evaluation of compounds

[0231] 1. CSF1R in vitro biochemical kinase assay

[0232] This invention uses the ADP-Glo™ kinase assay kit (Promega, cat. No. V9101) to quantify the amount of ADP generated during the kinase reaction, thereby evaluating the kinase inhibitory activity of the compounds. The specific experimental procedure is as follows: The kinase reaction was 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 of peptide; kinase reaction systems containing Hepes, MgCl2, Brij35, EGTA, and DTT were prepared to prepare 2× ATP and substrate solutions, and 2× kinase solutions, respectively; 20 nL of serially diluted compounds were transferred to the 384 assay plate using an Echo 655. Add 3 μL of 2× kinase solution to each well, mix well, and incubate at 25°C for 20 min. Add 3 μL of 2× substrate and ATP solution to each well and incubate at 25°C for 30 min. Add 4 μL of ADP-Glo ​​reagent to each well and incubate at 25°C for 40 min. Add 6 μL of Kinase Detection Reagent to each well and incubate at 25°C for 40 min. Read the RLU (Relative Luminescence Unit) signal using a BMG multi-function plate reader. The signal intensity was used to characterize the kinase activity, and the IC50 value of the compound was determined using a four-parameter curve in Graphpad Prism.

[0233] 2. In vitro biochemical kinase assays of C-KIT / FLT3 / PDGFRα / PDGFRβ

[0234] The selectivity of compounds for tyrosine kinase targets (C-KIT, FLT3, PDGFRα, PDGFRβ) was detected using the HTRF KinEASE-TK kit. The specific experimental procedure is as follows: The kinase reactions performed in this invention were carried out in 384-well plates (Greiner, cat. No. 784075), containing 1 nM C-KIT, 6 μM ATP, and 1 μM TK substrate; containing 0.024 nM FLT3, 2 μM ATP, and 1 μM TK substrate; containing 0.12 nM PDGFRα (Carna, cat. No. 08-157), 0.5 μM ATP, and 1 μM TK substrate; containing 0.23 nM PDGFRβ, 1 μM ATP, and 1 μM TK substrate; a 1× kinase reaction system containing MgCl2, MnCl, SEB, and DTT was prepared to prepare 2× ATP and substrate solutions and 2× kinase solutions; 25 nL of the compound was transferred to a 384-well 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 solutions using detection buffer; add 5 μL of Kinase Detection Reagent to the wells, and incubate at 25 °C for 60 min; use a BMG multi-function plate reader to read the fluorescence signals at 620 nm (Cryptate) and 665 nm (XL665), and use a four-parameter curve in Graphpad Prism to determine the IC50 of the compound. 50 Values. The results are shown in Table 1.

[0235] Example 16 Cell activity evaluation of the compound

[0236] Mouse myeloid leukemia lymphocyte line M-NFS-60 (RPMI 1640 + 0.05 mM β-mercaptoethanol + 62 ng / mL CSF-1 + 10% FBS), human monocytic leukemia cell line THP-1 (RPMI 1640 + 10% FBS + 0.05 mM β-mercaptoethanol + 1% P / S), and mouse microglia BV2 (MEM + 1% NEAA + 10% FBS + 1% P / S) were all purchased from Wuhan Pronosei Life Sciences Co., Ltd., and maintained normal growth under culture conditions of 37℃ and 5% CO2. Logarithmic growth phase BV2, M-NFS-60, and THP-1 cells were cultured at 8 × 10⁶ cells per well. 3 3×10 4 4×10 4Cells were seeded into 96-well plates, with 100 μL of culture medium per well, and three replicates were set for each drug concentration. After seeding BV2 cells, they were cultured overnight at 37°C and 5% CO2. After cell attachment, CSF1R inhibitor was added for 72 h. Suspension cells M-NFS-60 and THP-1 cells were allowed to stand for an appropriate time before drug treatment, followed by incubation for 72 h. The Cell Titer-Glo Luminescent Cell Viability Assay (Promega, cat. No. G7571) was used.

[0237] Cell viability was assessed. Following the reagent instructions, CellTiter-Glo Buffer was thawed before detection and equilibrated with the lyophilized substrate powder to room temperature. The two were then thoroughly mixed to prepare the CellTiter-Glo Reagent assay. Simultaneously, the cell culture plate and its contents were equilibrated at room temperature. 100 μL of CellTiter-Glo Reagent was added to either cell-containing or cell-free complete culture medium. The mixture was stirred for 2 min using a shaker to induce cell lysis. The culture plate was then incubated at room temperature for 10 min to stabilize the luminescence signal. The RLU (Relative Luminescence Unit) signal was read using a microplate reader. Signal intensity was used to characterize cell viability, and the IC50 value of the compound was determined using a four-parameter curve in Graphpad Prism. The results are shown in Table 1.

[0238] Table 1 Results of compound enzyme activity and cell viability assays Note: "A" refers to the CSF1R kinase inhibitory activity IC50 of the compound. 50 Less than 100 nM; "B" indicates that the compound CSF1R has an inhibitory activity of 100 nM. <IC 50 <1000nM; "C" refers to the CSF1R kinase inhibitory activity IC50 of the compound. 50 If the value is greater than 1000nM, NT indicates that it has not been detected.

[0239] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No markings in the claims should be construed as limiting the scope of the claims.

Claims

1. A class of compounds of formula I or II, their stereoisomers, or pharmaceutically acceptable salts, hydrates, or solvates thereof, characterized in that, As shown in Equations I and II R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, difluoromethyl, methoxy, morpholino, azirmonomethylpiperazine, or azirmonomethylpyrazole; R2 is selected from hydrogen, chlorine, fluorine, C1-C3 alkyl, or halogen-substituted C1-C3 alkyl; R3 is selected from hydrogen, C1-C6 alkyl, C3-C5 cycloalkyl, deuterated methyl or halogen-substituted C1-C6 alkyl; R4 and R5 are each independently selected from hydrogen or C1-C6 alkyl groups, or R4 and R5 together with the carbon atoms they are connected to form C3-C6 cycloalkyl groups.

2. The compound according to claim 1, its stereoisomers, or its pharmaceutically acceptable salts, hydrates, or solvates, characterized in that, in, R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, morpholino, N-methylpiperazine, or N-methylpyrazole; R2 is selected from hydrogen, fluorine, or C1-C3 alkyl groups; R3 is selected from hydrogen, C1-C3 alkyl, C3-C5 cycloalkyl, deuterated methyl or fluorine-substituted C1-C3 alkyl; R4 and R5 are each independently selected from hydrogen, C1 to C3 alkyl groups, or R4 and R5 together with the carbon atoms they are connected to form C3 to C5 cycloalkyl groups.

3. The compound according to claim 1, its stereoisomers, or its pharmaceutically acceptable salts, hydrates, or solvates, characterized in that, in, R1 is selected from hydrogen, chlorine, fluorine, trifluoromethyl, morpholino, N-methylpiperazine, or N-methylpyrazole; R2 is selected from hydrogen, fluorine, or methyl; R3 is selected from hydrogen, C1-C3 alkyl, cyclopropyl or deuterated methyl; R4 and R5 are each independently selected from hydrogen and methyl, or R4 and R5 together with the carbon atom they are connected to form a C3-C4 cycloalkyl group.

4. The compound according to claim 1, its stereoisomers, or its pharmaceutically acceptable salts, hydrates, or solvates, characterized in that, The compound is: 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. The compound, its stereoisomer, or its pharmaceutically acceptable salt, hydrate, or solvate according to any one of claims 1 to 4, characterized in that, The compound is used in the preparation of drugs for treating cancer.

6. The compound, its stereoisomer, or its pharmaceutically acceptable salt, hydrate, or solvate according to any one of claims 1 to 4, characterized in that, The compound is used in the preparation of drugs for treating tumors, autoimmune diseases, neurodegenerative diseases, traumatic brain injury, metabolic diseases, or metastatic diseases.

7. The compound, its stereoisomer, or its pharmaceutically acceptable salt, hydrate, or solvate according to any one of claims 1 to 4, characterized in that, The compound is used 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 carcinoma, colorectal cancer, gastrointestinal stromal tumors, melanoma, mesothelioma, osteosarcoma, head and neck cancer, glioblastoma, leukemia, peritoneal malignant tumors, peripheral T-cell lymphoma, bone metastatic cancer, multiple myeloma, metastasis of primary tumor sites, myeloproliferative disorders, hyperproliferative disorders, metabolic disorders, rheumatoid arthritis, osteoarthritis, pigmented villonodular synovitis, multiple sclerosis, autoimmune nephritis, lupus, Crohn's disease, asthma, neurodegenerative diseases, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, osteoporosis, hypereosinophilic syndrome, mastocytosis or mast cell leukemia, graft-versus-host disease, and traumatic brain injury.

8. The compound of any one of claims 1 to 4, its stereoisomer, or its pharmaceutically acceptable salt, hydrate, or solvate, characterized in that, The compound is used in the preparation of drugs for treating tenosynovial giant cell tumor, multiple myeloma, glioblastoma, metastatic or bone metastatic cancers at primary tumor sites, metabolic diseases, graft-versus-host disease, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, and traumatic brain injury.

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

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