A hpk1 kinase inhibitor and preparation method and application thereof

Abstract

This invention relates to an HPK1 kinase inhibitor, its preparation method, and its application. The HPK1 kinase inhibitor has the structure shown in formula (Ⅰ): (Ⅰ). Compared with the prior art, the HPK1 kinase inhibitor of this invention can effectively inhibit HPK1 activity, IC50... 50 The values ​​all reached nanomolar concentration levels, which is expected to improve the body's own immune function and weaken the immune escape effect of tumor cells.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, and relates to an HPK1 kinase inhibitor, its preparation method and application. Background Technology

[0002] Surgical resection, radiotherapy, chemotherapy, and small-molecule targeted drugs are the main methods used to treat cancer. Unfortunately, for many forms of cancer or tumors, surgical resection is often not a feasible option, and radiotherapy and chemotherapy, while killing tumor cells, also damage healthy cells. Furthermore, the instability of the tumor cell genome promotes mutations in tumor cells, leading to rapid changes in the cancer genome and resistance to drugs specifically targeting tumors, making cancer treatment extremely difficult. In recent years, a novel strategy for cancer treatment has emerged: leveraging the patient's own immune system to kill tumor cells and enhancing the body's anti-tumor immunity. One approach involves inhibiting negative regulators of the immune response that maintain peripheral tolerance, causing the tumor to be recognized as a non-self antigen, thereby overcoming the immune escape of tumor cells.

[0003] Hematopoietic progenitor cell kinase (HPK1) is a member of the mitogen-activated protein kinase kinase kinase (MAP4K) family of Ste20 serine / threonine kinases, also known as MAP4K1. It is primarily expressed in hematopoietic cells (such as T cells, B cells, neutrophils, dendritic cells, and macrophages), playing a negative regulatory role in T and B cells. Inhibition of HPK1 kinase activity can enhance the ability of immune cells to kill tumor cells, making HPK1 a highly valuable target in tumor immunology. HPK1 kinase is not expressed in major organs, suggesting that HPK1 kinase inhibitors may not cause any serious complications. Summary of the Invention

[0004] The purpose of this invention is to provide an HPK1 kinase inhibitor with a novel structure and high activity, as well as its preparation method and application.

[0005] The objective of this invention can be achieved through the following technical solutions: One of the technical solutions of the present invention is to provide an HPK1 kinase inhibitor, which has the structure shown in formula (Ⅰ) below: (I), R1 is selected from H and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group, nitrogen-containing cyclohexane group containing 0-3 substituents, and any combination thereof, wherein the substituents are selected from any combination thereof, methyl, amino, or trifluoromethyl; R2 is selected from H and C. 1-6Alkyl, C 1-6 Alkoxy, C 3-8 The nitrogen-containing bicyclic heterocyclic group comprises any one or more combinations of cycloalkyl, phenyl, phenyl containing N-methylacetamide substituents, and nitrogen-containing bicyclic heterocyclic groups, wherein the nitrogen-containing heterocycle in the nitrogen-containing bicyclic heterocyclic group has no substituents or contains methyl substituents, and the ring carbon has no substituents or contains fluorine substituents or contains methoxy substituents.

[0006] In some specific embodiments, R1 is selected from... , , , Any one of them; R2 is selected from , , , Any one of them.

[0007] R1 is connected to the imidazole ring and consists of a carbon chain of two carbon atoms; R2 is connected to the amino group on the triazine ring and consists of a nitrogen atom and a carbon atom, forming a carbon-nitrogen bond.

[0008] In some specific embodiments, the HPK1 kinase inhibitor is selected from any one of the following structural formulas: , , , , , , , , , , , , .

[0009] The second technical solution of the present invention is to provide a method for preparing an HPK1 kinase inhibitor as described in one of the above technical solutions, comprising the following steps: S1. Dissolve guanidine hydrochloride and 2-chloro-4-nitroimidazole in tetrahydrofuran, replace with nitrogen, slowly add tetrahydrofuran solution containing potassium tert-butoxide, and react the resulting mixture under nitrogen protection to obtain intermediate 1; S2. Dissolve intermediate 1 obtained in step S1 in a mixed solution of dioxane and water, add compound A, Pd(dppf)Cl2 and sodium carbonate, and react under nitrogen protection to obtain intermediate 2. S3. Dissolve intermediate 2 obtained in step S2 in a mixed solution of toluene and tetrahydrofuran, add compound B, cuprous iodide, N,N'-dimethylethylenediamine and potassium phosphate, and react under nitrogen protection to obtain intermediate 3. S4. Dissolve intermediate 3 obtained in step S3 in dichloromethane solution, add trifluoroacetic acid, stir the reaction to obtain the target compound, which is the HPK1 kinase inhibitor. in, The structural formula of compound A is: , R3 is selected from hydrogen, halogen, nitro, cyano, and C. 1-6 Alkyl, C 1-6 Any combination of one or more of the following: alkyl halogroups, hydroxyl groups, mercapto groups, and amino groups; R4 is selected from hydrogen, halogen, nitro, cyano, and C. 1-6 Alkyl, C 1-6 Any combination of one or more of the following: alkyl halogroups, hydroxyl groups, mercapto groups, and amino groups; X1 is selected from either C or N; The structural formula of compound B is: , R5 is selected from hydrogen, halogen, cyano, nitro, and -OR. x -NHR x -N(R) x )2、-C(O)R x C 1-6 Alkyl, C 1-6 Any one or more combinations of haloalkyl groups; The R x Selected from hydrogen, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Any one of the haloalkyl groups; wherein, the C 1-6 Hydroxyalkyl groups without substituents or containing oxy or C groups 1-3 Alkyl, phenyl, or any one or more substituent groups; R6 is selected from hydrogen, halogen, cyano, hydroxyl, C 1-6 Any one of alkyl, 3-10 membered carbon cyclic group, oxygen-containing heterocyclic group, and nitrogen-containing heterocyclic group; wherein, the C 1-6 Alkyl, 3-10 membered carbon cyclic group, oxygen-containing heterocyclic group, nitrogen-containing heterocyclic group without substituents or containing 1-4 R7 substituents; The R7 is selected from hydrogen, halogen, hydroxyl, amide, C 1-6 Any combination of one or more alkyl groups; X2 is selected from either C or N; X3 is selected from either C or N.

[0010] In some specific embodiments, the structure of compound A is selected from any of the following structural formulas: They are named as follows: tert-butyl (tert-butoxycarbonyl) (5-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)pyridin-3-yl)carbamate, tert-butyl (tert-butoxycarbonyl) (4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)pyridin-3-yl)carbamate, tert-butyl (tert-butoxycarbonyl) (2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)pyridin-3-yl)carbamate, tert-butyl (tert-butoxycarbonyl) (5-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-4-(trifluoromethyl)pyridin-2-yl)carbamate; The structure of compound B is selected from any of the following structural formulas: They were named as follows: 7-iodo-6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline, 6-fluoro-7-iodo-2-methyl-1,2,3,4-tetrahydroisoquinoline, 7-iodo-2-methyl-1,2,3,4-tetrahydroisoquinoline, and 2-(4-iodophenyl)-N-methylacetamide.

[0011] In some specific embodiments, in step S1, the molar ratio of 2-chloro-4-nitroimidazole, guanidine hydrochloride, and potassium tert-butoxide is 1: (2~3): (6~7). The reaction was carried out under nitrogen atmosphere at 50-80℃ with stirring for 10-20 h.

[0012] As a more preferred embodiment, in step S1, the molar ratio of 2-chloro-4-nitroimidazole, guanidine hydrochloride, and potassium tert-butoxide is 1:2:6.

[0013] In some specific embodiments, in step S2, the molar ratio of intermediate 1, compound A, Pd(dppf)Cl2, and sodium carbonate is 1:(2~3):(0.1~0.2):(3~4). The reaction was carried out under nitrogen atmosphere at 70-120℃ with stirring for 1-7 h.

[0014] As a more preferred embodiment, in step S2, the molar ratio of intermediate 1, compound A, Pd(dppf)Cl2, and sodium carbonate is 1:2:0.1:3.

[0015] In some specific embodiments, in step S3, the molar ratio of intermediate 2, compound B, cuprous iodide, N,N'-dimethylethylenediamine, and potassium phosphate is 1:1:1:1:(3~4). Under nitrogen atmosphere, react at 80-120℃ for 20-30 h with stirring.

[0016] As a more preferred embodiment, in step S3, the molar ratio of intermediate 2, compound B, cuprous iodide, N,N'-dimethylethylenediamine, and potassium phosphate is 1:1:1:1:3.

[0017] In some specific embodiments, in step S4, the molar ratio of intermediate 3 to trifluoroacetic acid is 1:(1.5~1.6). The reaction was stirred at room temperature for 1-10 h at a temperature of 15℃-25℃.

[0018] As a more preferred embodiment, in step S4, the molar ratio of intermediate 3 to trifluoroacetic acid is 1:1.5.

[0019] The third technical solution of the present invention is to provide the use of the HPK1 kinase inhibitor as described in one of the above technical solutions in the preparation of a drug for the prevention and / or treatment of HPK1 inhibition-related lung cancer, melanoma, liver cancer, kidney cancer, prostate cancer, pancreatic cancer, rectal cancer, colon cancer, breast cancer, bladder cancer, and gastric cancer.

[0020] Compared with the prior art, the present invention has the following advantages: (1) The method for synthesizing the HPK1 kinase inhibitor of the present invention is simple, the raw materials are readily available, and it can be obtained through a few simple reaction steps.

[0021] (2) The HPK1 kinase inhibitor of the present invention can effectively inhibit HPK1 activity, and is expected to improve the body's own immune function and weaken the immune escape effect of tumor cells. The HPK1 kinase inhibitor IC prepared by the present invention... 50 All values ​​reached nanomolar concentration levels, compared to PF-07265028 currently in Phase I clinical trials, IC50 values ​​were significantly higher. 50 The values ​​are on the same order of magnitude or even lower, which shows that the HPK1 kinase inhibitor of the present invention has broad application prospects in the preparation of drugs for treating and / or preventing tumors related to HPK1 kinase inhibition. Attached Figure Description

[0022] Figure 1 This is a flowchart illustrating the synthesis process of the HPK1 kinase inhibitor of this invention. Detailed Implementation

[0023] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. These embodiments are based on the technical solution of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0025] Unless otherwise specified, the materials and processes described in the following embodiments or examples are conventional materials and processes used in the art to achieve the corresponding functions.

[0026] Example 1: The compound synthesized in this embodiment is named N-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)-6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline-7-amine, and its structural formula is as follows: .

[0027] It includes the following preparation steps: Step a: At room temperature (25℃), a solution of tetrahydrofuran (100 mL) containing potassium tert-butoxide (22.82 g, 203.37 mmol) was slowly added dropwise to a solution of tetrahydrofuran (100 mL) containing 2-chloro-4-nitroimidazole (5.00 g, 33.89 mmol) and guanidine hydrochloride (6.48 g, 67.79 mmol). The mixture was heated to 65℃ and refluxed under nitrogen protection for 12 h. After the reaction was completed, the mixture was cooled to room temperature (25℃), and a saturated ammonium chloride solution was added. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1.69 g of intermediate 1, with a yield of 29.2%. Its structural formula is as follows: .

[0028] Step b: Intermediate 1 (1 g, 5.86 mmol) was dissolved in a mixed solution of dioxane (21 mL) and water (3 mL). Compound A (5.73 g, 11.73 mmol), Pd(dppf)Cl2 (429 mg, 0.59 mmol), and sodium carbonate (1.86 g, 17.59 mmol) were added. The mixture was refluxed at 90 °C for 5 h under nitrogen protection, cooled to room temperature (25 °C), filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain 2.36 g of intermediate 2, with a yield of 81.1%. Its structural formula is as follows: .

[0029] Compound A is tert-butyl (tert-butoxycarbonyl) (5-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-4-(trifluoromethyl)pyridin-2-yl)carbamate, and its structural formula is as follows: .

[0030] Step c: Intermediate 2 (1 g, 2.01 mmol) was dissolved in a mixed solution of toluene (20 mL) and tetrahydrofuran (4 mL). Compound B (0.61 g, 2.01 mmol), cuprous iodide (0.38 g, 2.01 mmol), N,N'-dimethylethylenediamine (0.18 g, 2.01 mmol), and potassium phosphate (1.28 g, 6.04 mmol) were added. The mixture was refluxed at 110 °C for 24 h under nitrogen protection. After cooling to room temperature (25 °C), the mixture was filtered through diatomaceous earth. The filtrate was evaporated to dryness under reduced pressure and purified by column chromatography to give 580 mg of intermediate 3, with a yield of 43.0%. Its structural formula is as follows: .

[0031] Compound B is 7-iodo-6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline, and its structural formula is as follows: .

[0032] Step d: Trifluoroacetic acid (92 mg, 1.3 mmol) was added to a solution of intermediate 3 (580 mg, 0.86 mmol) in dichloromethane (5 mL), and the mixture was stirred at room temperature (25 °C) for 2 h. The pH was adjusted to neutral by adding saturated sodium bicarbonate, and the mixture was extracted three times with dichloromethane. The organic phases were combined, evaporated under reduced pressure, and purified by column chromatography to give 291 mg of the target compound, with a yield of 71.5%.

[0033] The high-resolution mass spectrometry data of the target compound N-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)-6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline-7-amine are as follows (MS (ESI): M / Z = 472.1743 [M+H] + .

[0034] 1H NMR (401 MHz, DMSO-d6): δ 9.33 (s, 1H), 9.02 (s, 1H), 7.50 (s, 1H), 7.14 (t, J =1.0 Hz, 1H), 6.77 (t, J = 1.0 Hz, 1H), 6.34 (s, 2H), 3.88 (s, 3H), 3.64 (d, J= 1.0 Hz, 2H), 2.85-2.64 (m, 5H), 2.35 (s, 3H). 13 C NMR (101 MHz, DMSO-d6): δ160.39, 160.36, 160.34, 160.31, 156.04, 155.32, 155.30, 155.28, 149.90,149.88, 149.86, 149.86, 147.84, 141.79, 141.69, 131.04, 129.73, 129.08,128.77, 128.45, 128.14, 127.75, 127.64, 124.97, 122.33, 119.66, 118.87,113.07, 112.20, 112.16, 112.15, 112.08, 105.68, 105.65, 105.60, 105.56, 57.54, 55.96, 52.25, 44.59, 27.62. Example 2: The compound synthesized in this embodiment is named N-(6-(5-amino-6-methylpyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)-6-fluoro-2-methyl-1,2,3,4-tetrahydroisoquinoline-7-amine, and its structural formula is as follows: .

[0035] It includes the following preparation steps: Step a: At room temperature (25℃), a solution of potassium tert-butoxide (22.82 g, 203.37 mmol) in tetrahydrofuran (100 mL) was slowly added dropwise to a solution of 2-chloro-4-nitroimidazole (5.00 g, 33.89 mmol) and guanidine hydrochloride (6.48 g, 67.79 mmol) in tetrahydrofuran (100 mL). The mixture was heated to 65℃ and refluxed under nitrogen protection for 12 h. After the reaction was completed, the mixture was cooled to room temperature, and a saturated ammonium chloride solution was added. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1.69 g of intermediate 1, with a yield of 29.2%. Its structural formula is as follows: .

[0036] Step b: Intermediate 1 (1.00 g, 5.86 mmol) was dissolved in a mixed solution of dioxane (21 mL) and water (3 mL). Compound A (5.09 g, 11.73 mmol), Pd(dppf)Cl2 (429 mg, 0.59 mmol), and sodium carbonate (1.86 g, 17.59 mmol) were added. The mixture was refluxed at 90 °C for 5 h under nitrogen protection, cooled to room temperature (25 °C), filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain 2.26 g of intermediate 2, with a yield of 87.3%. Its structural formula is as follows: .

[0037] Compound A is tert-butyl (tert-butoxycarbonyl) (2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)pyridin-3-yl)carbamate, with the following structural formula: .

[0038] Step c: Intermediate 2 (1 g, 2.26 mmol) was dissolved in a mixed solution of toluene (20 mL) and tetrahydrofuran (4 mL). Compound B (0.66 g, 2.26 mmol), cuprous iodide (0.43 g, 2.26 mmol), N,N'-dimethylethylenediamine (0.20 g, 2.26 mmol), and potassium phosphate (1.44 g, 6.78 mmol) were added. The mixture was refluxed at 110 °C for 24 h under nitrogen protection. After cooling to room temperature (25 °C), the mixture was filtered through diatomaceous earth. The filtrate was evaporated to dryness under reduced pressure and purified by column chromatography to obtain 519 mg of intermediate 3, with a yield of 37.9%. Its structural formula is as follows: .

[0039] Compound B is 6-fluoro-7-iodo-2-methyl-1,2,3,4-tetrahydroisoquinoline, and its structural formula is as follows: .

[0040] Step d: Trifluoroacetic acid (93 mg, 1.29 mmol) was added to a 5 mL solution of intermediate 3 (519 mg, 0.86 mmol) in dichloromethane, and the mixture was stirred at room temperature for 2 h. The pH was adjusted to neutral by adding saturated sodium bicarbonate, and the mixture was extracted three times with dichloromethane. The organic phases were combined, evaporated to dryness under reduced pressure, and purified by column chromatography to give 216 mg of the target compound, with a yield of 62.2%.

[0041] The high-resolution mass spectrometry data of the target compound N-(6-(5-amino-6-methylpyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)-6-fluoro-2-methyl-1,2,3,4-tetrahydroisoquinoline-7-amine are as follows: MS (ESI): M / Z = 406.1826 [M+H] + .

[0042] 1 H NMR (401 MHz, DMSO-d6): δ 9.89 (s, 1H), 8.79 (d, J = 1.6 Hz, 1H), 7.78 (d, J = 1.8 Hz, 1H), 7.17 (dt, J = 4.9, 1.0 Hz, 1H), 6.95 (dt, J = 8.0, 0.9Hz, 1H), 4.98 (s, 2H), 3.62 (d, J = 0.9 Hz, 2H), 2.92 - 2.77 (m, 4H), 2.34(s, 3H), 2.28 (s, 3H). 13 C NMR (101 MHz, DMSO-d6): δ 156.02, 155.91, 153.92,153.17, 150.68, 145.84, 142.64, 141.55, 141.45, 138.36, 130.89, 130.81,129.67, 129.65, 128.67, 128.46, 126.46, 119.45, 119.38, 118.59, 115.79,115.59, 57.40, 52.14, 44.63, 27.84, 27.82, 18.77. Example 3: The compound synthesized in this embodiment is named N-(6-(5-amino-4-methylpyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)-2-methyl-1,2,3,4-tetrahydroisoquinoline-7-amine, and its structural formula is as follows: .

[0043] It includes the following preparation steps: Step a: At room temperature (25℃), a solution of potassium tert-butoxide (22.82 g, 203.37 mmol) in tetrahydrofuran (100 mL) was slowly added dropwise to a solution of 2-chloro-4-nitroimidazole (5.00 g, 33.89 mmol) and guanidine hydrochloride (6.48 g, 67.79 mmol) in tetrahydrofuran (100 mL). The mixture was heated to 65℃ and refluxed under nitrogen protection for 12 h. After the reaction was completed, the mixture was cooled to room temperature, and a saturated ammonium chloride solution was added. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1.69 g of intermediate 1, with a yield of 29.2%. Its structural formula is as follows: .

[0044] Step b: Intermediate 1 (1 g, 5.86 mmol) was dissolved in a mixed solution of dioxane (21 mL) and water (3 mL). Compound A (5.09 g, 11.73 mmol), Pd(dppf)Cl2 (429 mg, 0.59 mmol), and sodium carbonate (1.86 g, 17.59 mmol) were added. The mixture was refluxed at 90 °C for 5 h under nitrogen protection, cooled to room temperature (25 °C), filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain 2.09 g of intermediate 2, with a yield of 80.7%. Its structural formula is as follows: .

[0045] Compound A is a tert-butyl (tert-butoxycarbonyl) (4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)pyridin-3-yl) carbamate, with the following structural formula: .

[0046] Step c: Intermediate 2 (1 g, 2.26 mmol) was dissolved in a mixed solution of toluene (20 mL) and tetrahydrofuran (4 mL). Compound B (0.62 g, 2.26 mmol), cuprous iodide (0.43 g, 2.26 mmol), N,N'-dimethylethylenediamine (0.20 g, 2.26 mmol), and potassium phosphate (1.44 g, 6.78 mmol) were added. The mixture was refluxed at 110 °C for 24 h under nitrogen protection. After cooling to room temperature (25 °C), the mixture was filtered through diatomaceous earth. The filtrate was evaporated to dryness under reduced pressure and purified by column chromatography to give 631 mg of intermediate 3, with a yield of 47.4%. Its structural formula is as follows: .

[0047] Compound B is 7-iodo-2-methyl-1,2,3,4-tetrahydroisoquinoline, and its structural formula is as follows: .

[0048] Step d: Trifluoroacetic acid (116 mg, 1.61 mmol) was added to a solution of intermediate 3 (631 mg, 1.07 mmol) in dichloromethane (5 mL), and the mixture was stirred at room temperature for 2 h. The pH was adjusted to neutral by adding saturated sodium bicarbonate, and the mixture was extracted three times with dichloromethane. The organic phases were combined, evaporated to dryness under reduced pressure, and purified by column chromatography to give 305 mg of the target compound, with a yield of 73.3%.

[0049] The high-resolution mass spectrometry data of the target compound N-(6-(5-amino-4-methylpyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)-2-methyl-1,2,3,4-tetrahydroisoquinoline-7-amine are as follows (MS(ESI): M / Z = 388.1920 [M+H] + .

[0050] 1 H NMR (401 MHz, DMSO-d6): δ 8.99 (s, 1H), 8.90 (d, J = 1.6 Hz, 1H), 8.12 (d, J = 1.8 Hz, 1H), 7.30 (dd, J = 7.7, 2.2 Hz, 1H), 7.26 (dt, J = 2.2, 1.0Hz, 1H), 7.06 (dt, J = 7.6, 1.0 Hz, 1H), 5.07 (s, 2H), 3.58 (d, J = 1.0 Hz,2H), 2.97 - 2.86 (m, 2H), 2.84 - 2.66 (m, 2H), 2.34 (s, 3H). 13 C NMR (101 MHz, DMSO-d6): δ 156.16, 152.87, 142.94, 141.78, 141.75, 138.92, 137.64, 137.46, 132.58, 130.38, 127.73, 127.57, 123.82, 118.42, 118.01, 57.78, 52.38, 44.66,28.19, 13.23. Example 4: The compound synthesized in this embodiment is named 2-(4-((6-(5-aminopyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)amino)phenyl)-N-methylacetamide, and its structural formula is as follows: .

[0051] It includes the following preparation steps: Step a: At room temperature (25℃), a solution of potassium tert-butoxide (22.82 g, 203.37 mmol) in tetrahydrofuran (100 mL) was slowly added dropwise to a solution of 2-chloro-4-nitroimidazole (5.00 g, 33.89 mmol) and guanidine hydrochloride (6.48 g, 67.79 mmol) in tetrahydrofuran (100 mL). The mixture was heated to 65℃ and refluxed under nitrogen protection for 12 h. After the reaction was completed, the mixture was cooled to room temperature, and a saturated ammonium chloride solution was added. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1.69 g of intermediate 1, with a yield of 29.2%. Its structural formula is as follows: .

[0052] Step b: Intermediate 1 (1 g, 5.86 mmol) was dissolved in a mixed solution of dioxane (21 mL) and water (3 mL), and compound A (4.93 g, 11.73 mmol), Pd(dppf)Cl2 (429 mg, 0.59 mmol), and sodium carbonate (1.86 g, 17.59 mmol) were added. The mixture was refluxed at 90 °C for 5 h under nitrogen protection, cooled to room temperature (25 °C), filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain 2.02 g of intermediate 2, with a yield of 80.5%. Its structural formula is as follows: .

[0053] Compound A is tert-butyl (tert-butoxycarbonyl) (5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)pyridin-3-yl)carbamate, with the following structural formula: .

[0054] Step c: Intermediate 2 (1 g, 2.33 mmol) was dissolved in a mixed solution of toluene (20 mL) and tetrahydrofuran (4 mL). Compound B (0.64 g, 2.33 mmol), cuprous iodide (0.44 g, 2.33 mmol), N,N'-dimethylethylenediamine (0.21 g, 2.33 mmol), and potassium phosphate (1.28 g, 7.00 mmol) were added. The mixture was refluxed at 110 °C for 24 h under nitrogen protection. After cooling to room temperature (25 °C), the mixture was filtered through diatomaceous earth. The filtrate was evaporated to dryness under reduced pressure and purified by column chromatography to give 602 mg of intermediate 3, with a yield of 44.9%. Its structural formula is as follows: .

[0055] Compound B is 2-(4-iodophenyl)-N-methylacetamide, and its structural formula is as follows: .

[0056] Step d: Trifluoroacetic acid (113 mg, 1.57 mmol) was added to a solution of intermediate 3 (602 mg, 1.05 mmol) in dichloromethane (5 mL), and the mixture was stirred at room temperature for 2 h. The pH was adjusted to neutral by adding saturated sodium bicarbonate, and the mixture was extracted three times with dichloromethane. The organic phases were combined, evaporated to dryness under reduced pressure, and purified by column chromatography to give 298 mg of the target compound, with a yield of 75.9%.

[0057] The high-resolution mass spectrometry data of the target compound 2-(4-((6-(5-aminopyridin-3-yl)-7H-imidazo[4,5-e][1,2,4]triazin-3-yl)amino)phenyl)-N-methylacetamide are as follows: MS (ESI): M / Z = 376.1556 [M+H] + .

[0058] 1 H NMR (401 MHz, DMSO-d6): δ 9.14 (t, J = 1.7 Hz, 1H), 8.76 (s, 1H), 8.23 ​​(t, J = 1.7 Hz, 1H), 7.99 (t, J = 1.7 Hz, 1H), 7.79 (q, J = 4.9 Hz, 1H), 7.49– 7.42 (m, 2H), 7.22 (dt, J = 7.8, 1.0 Hz, 2H), 5.48 (s, 2H), 3.46 (t, J =1.0 Hz, 2H), 2.70 (d, J = 4.7 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6): δ 171.08,155.70, 152.87, 142.61, 142.58, 141.65, 141.60, 139.10, 137.08, 129.95,129.67, 128.80, 119.65, 117.23, 41.78, 26.43. Comparative Example 1: Most of them are the same as in Example 1, except for the equivalent of potassium tert-butoxide.

[0059] In step a: At room temperature (25°C), a solution of potassium tert-butoxide (2.28 g, 20.34 mmol) in tetrahydrofuran (100 mL) was slowly added dropwise to a solution of 2-chloro-4-nitroimidazole (1.00 g, 6.78 mmol) and guanidine hydrochloride (1.30 g, 13.56 mmol) in tetrahydrofuran (20 mL). The mixture was heated to 65°C and refluxed under nitrogen protection for 12 h. After the reaction was completed, the mixture was cooled to room temperature, and a saturated ammonium chloride solution was added. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 140 mg of intermediate 1, with a yield of 12.1%. Its structural formula is as follows: .

[0060] Potassium tert-butoxide acts as a strong base to activate guanidine hydrochloride, and a decrease in the equivalent amount of potassium tert-butoxide in the reaction will significantly reduce the yield of the target product.

[0061] Comparative Example 2: Most of the contents are the same as in Example 1, except for the equivalent of Pd(dppf)Cl2.

[0062] In step b: the product from step a (1 g, 5.86 mmol) was dissolved in a mixed solution of dioxane (21 mL) and water (3 mL), and tert-butyl (tert-butyloxycarbonyl) (5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)-4-(trifluoromethyl)pyridin-2-yl)carbamate (5.73 g, 11.73 mmol), Pd(dppf)Cl2 (214 mg, 0.29 mmol), and sodium carbonate (1.86 g, 17.59 mmol) were added. The mixture was refluxed at 90 °C for 5 h under nitrogen protection, cooled to room temperature, filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain 0.91 g of intermediate 2, with a yield of 31.2%. Its structural formula is as follows: .

[0063] As a catalyst for the Suzuki coupling reaction, the yield of the target product decreases significantly when the amount of Pd(dppf)Cl2 decreases.

[0064] The compounds prepared in Examples 1-4 were subjected to HPK1 protein inhibitory activity tests, and their IC50 values ​​were... 50 The measurement results are as follows: This invention utilizes model predictions from computer-aided drug design to design and synthesize a series of compounds containing 1,2,4-imidazotriazine fragments. HPK1 kinase inhibitor activity assays revealed that this series of compounds exhibited IC50 levels of the same or lower than those of PF-07265028, which showed excellent efficacy in Phase I clinical trials. 50 The test results hold promise for use in the development of drugs to treat cancer and other diseases related to HPK1 activity.

[0065] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. An HPK1 kinase inhibitor, characterized in that, The structure is as shown in equation (Ⅰ): （Ⅰ）， R1 is selected from H and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group, nitrogen-containing cyclohexane group containing 0-3 substituents, and any combination thereof, wherein the substituents are selected from any combination thereof, methyl, amino, or trifluoromethyl; R2 is selected from H and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The nitrogen-containing bicyclic heterocyclic group comprises any one or more combinations of cycloalkyl, phenyl, phenyl containing N-methylacetamide substituents, and nitrogen-containing bicyclic heterocyclic groups, wherein the nitrogen-containing heterocycle in the nitrogen-containing bicyclic heterocyclic group has no substituents or contains methyl substituents, and the ring carbon has no substituents or contains fluorine-substituted O groups or contains methoxy substituents.

2. The HPK1 kinase inhibitor according to claim 1, characterized in that, R1 is selected from , , , Any one of them; R2 is selected from , , , Any one of them.

3. The HPK1 kinase inhibitor according to claim 1, characterized in that, The HPK1 kinase inhibitor is selected from any one of the following structural formulas: 、 、 、 、 、 、 、 、 、 、 、 、 。 4. A method for preparing an HPK1 kinase inhibitor as described in any one of claims 1 to 3, characterized in that, Includes the following steps: S1. Dissolve guanidine hydrochloride and 2-chloro-4-nitroimidazole in tetrahydrofuran, replace with nitrogen, slowly add tetrahydrofuran solution containing potassium tert-butoxide, and react the resulting mixture under nitrogen protection to obtain intermediate 1; S2. Dissolve intermediate 1 obtained in step S1 in a mixed solution of dioxane and water, add compound A, Pd(dppf)Cl2 and sodium carbonate, and react under nitrogen protection to obtain intermediate 2. S3. Dissolve intermediate 2 obtained in step S2 in a mixed solution of toluene and tetrahydrofuran, add compound B, cuprous iodide, N,N'-dimethylethylenediamine and potassium phosphate, and react under nitrogen protection to obtain intermediate 3. S4. Dissolve intermediate 3 obtained in step S3 in dichloromethane solution, add trifluoroacetic acid, stir the reaction to obtain the target compound, which is the HPK1 kinase inhibitor. in, The structural formula of compound A is: , R3 is selected from hydrogen, halogen, nitro, cyano, and C. 1-6 Alkyl, C 1-6 Any combination of one or more of the following: alkyl halogroups, hydroxyl groups, mercapto groups, and amino groups; R4 is selected from hydrogen, halogen, nitro, cyano, and C. 1-6 Alkyl, C 1-6 Any combination of one or more of the following: alkyl halogroups, hydroxyl groups, mercapto groups, and amino groups; X1 is selected from either C or N; The structural formula of compound B is: , R5 is selected from hydrogen, halogen, cyano, nitro, and -OR. x -NHR x -N(R) x )2、-C(O)R x C 1-6 Alkyl, C 1-6 Any one or more combinations of haloalkyl groups; The R x Selected from hydrogen, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Any one of the haloalkyl groups; wherein, the C 1-6 Hydroxyalkyl groups without substituents or containing oxy or C groups 1-3 Alkyl, phenyl, or any one or more substituent groups; R6 is selected from hydrogen, halogen, cyano, hydroxyl, C 1-6 Any one or more combinations of alkyl, 3-10 membered carbon cyclic groups, oxygen-containing heterocyclic groups, and nitrogen-containing heterocyclic groups; wherein, the C 1-6 Alkyl, 3-10 membered carbon cyclic, oxygen-containing heterocyclic, nitrogen-containing heterocyclic without substituents or containing 1-4 R7 substituents respectively; The R7 is selected from hydrogen, halogen, hydroxyl, amide, C 1-6 Any combination of one or more alkyl groups; X2 is selected from either C or N; X3 is selected from either C or N.

5. The method for preparing the HPK1 kinase inhibitor according to claim 4, characterized in that, The structure of compound A is selected from any one of the following structural formulas: The structure of compound B is selected from any one of the following structural formulas: 。 6. The method for preparing the HPK1 kinase inhibitor according to claim 4, characterized in that, In step S1, the molar ratio of 2-chloro-4-nitroimidazole, guanidine hydrochloride, and potassium tert-butoxide is 1:(2~3):(6~7). The reaction was carried out under nitrogen atmosphere at 50-80℃ with stirring for 10-20 h.

7. The method for preparing the HPK1 kinase inhibitor according to claim 4, characterized in that, In step S2, the molar ratio of intermediate 1, compound A, Pd(dppf)Cl2, and sodium carbonate is 1:(2~3):(0.1~0.2):(3~4). The reaction was carried out under nitrogen atmosphere at 70-120℃ with stirring for 1-7 h.

8. The method for preparing the HPK1 kinase inhibitor according to claim 4, characterized in that, In step S3, the molar ratio of intermediate 2, compound B, cuprous iodide, N,N'-dimethylethylenediamine, and potassium phosphate is 1:1:1:1:(3~4). Under nitrogen atmosphere, react at 80-120℃ for 20-30 h with stirring.

9. The method for preparing the HPK1 kinase inhibitor according to claim 4, characterized in that, In step S4, the molar ratio of intermediate 3 to trifluoroacetic acid is 1:(1.5~1.6). The reaction was stirred at room temperature for 1-10 h at a temperature of 15℃-25℃.

10. The use of an HPK1 kinase inhibitor as described in any one of claims 1 to 3 in the preparation of a medicament for the prevention and / or treatment of HPK1 inhibition-related tumors, wherein the tumors are selected from lung cancer, melanoma, liver cancer, kidney cancer, Any one of the following: prostate cancer, pancreatic cancer, rectal cancer, colon cancer, breast cancer, bladder cancer, or stomach cancer.

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