1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compounds and their preparation and use

Abstract

The present invention belongs to the technical field of medicinal chemistry. Specifically, the present invention discloses a 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound and its preparation and use. The compound of the present invention has a structure represented by formula (I). The compound has a novel structure and is a kind of adenosine receptor inhibitor. It has excellent inhibitory activity against adenosine A2A receptor and good subtype selectivity. It can inhibit adenosine A2A receptor and has adenosine A2A receptor and ... 2A Adenosine A is used in immunotherapy as a receptor-targeting inhibitor. 2A By inhibiting the activity of the receptor, the immune system can release the tumor cell rejection function, thereby playing a role in treating tumors. At the same time, the compound of the present invention is easy to prepare, uses inexpensive compounds as raw materials, has mild reaction conditions, has high total yield and purity, and can be produced on a scale of several tens of grams, making it suitable for use in the subsequent clinical trials of adenosine A 2A This will be useful in researching potential receptor inhibitors. [C70] TIFF2025503599000074.tif35170

Description

[Technical Field]

[0001] The present invention belongs to the technical field of medicinal chemistry, and particularly relates to 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compounds and their preparation and use. [Background technology]

[0002] The adenosine receptor is a G protein-coupled receptor, a membrane protein containing seven transmembrane α-helices, A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15, A16, A17, A18, A19, A19, A20, A21, A22, A23, A24, A25, 2A , A 2B There are four subtypes of adenosine receptors: A, A2, and A3. The tumor microenvironment contains a large amount of extracellular adenosine, and high concentrations of extracellular adenosine interact with G protein-coupled adenosine receptors on the membrane surface of immune cells, transmitting immunosuppressive signals into the immune cells, suppressing the proliferation and maturation of immune cells such as T cells, NK cells, macrophages, and dendritic DC cells, and inhibiting the production of cytokines (IL-2, IFN-g) by immune cells, leading to immune impairment and tumor cell immune escape. Of the four types of G protein-coupled adenosine receptors, adenosine A 2A Adenosine receptors are the most abundant and widely expressed subtype in immune cells such as T cells and NK cells, and when they bind to adenosine, they exert immunosuppressive effects. Therefore, the development of adenosine receptor inhibitors is of great significance for improving the immune function of the body.

[0003] Currently, small molecule adenosine receptor inhibitors are being widely studied, and A 2A Blocking the transmission of immunosuppressive signals via receptors can improve the body's immune response. 2A Receptor inhibitors are already in clinical research, and these small molecule adenosine receptor inhibitors have remarkable therapeutic effects on common tumor diseases such as melanoma, lung squamous cell carcinoma, breast cancer, gastric cancer, ovarian cancer, and prostate cancer. However, the development of small molecule adenosine receptor inhibitors is still in its early stages, and there is much room for improvement. Therefore, research into novel adenosine receptor inhibitors with more anticancer activity is urgently needed. Summary of the Invention

[0004] To overcome the above-mentioned shortcomings of the existing technology, the first object of the present invention is to provide 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compounds.

[0005] A second object of the present invention is to provide a method for producing the above 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound.

[0006] The third object of the present invention is to provide a use of the above-mentioned 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound, which is capable of inhibiting adenosine A. 2A It has excellent inhibitory activity against the A receptor and good subtype selectivity. 2A It is expected to be used in tumor immunotherapy that targets receptors.

[0007] The first object of the present invention is achieved by the following technical measures.

[0008] A 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound having the structure represented by formula (I): [ka] However, R 1 are independently selected from hydrogen and C1-C4 alkyl groups; R 2 are independently a C1-C6 alkyl group, a C1-C4 alkanol group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, a substituted phenyl group, 2-pyridine, 3-pyridine, 4-pyridine, 2-furan, a C4-C12 heterocycle, [ka] Selected from.

[0009] R 4are independently selected from a substituted phenyl group, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, imidazole, pyrazole, oxazole, pyrazine, pyridazine, 2-furan, 2-substituted furan, thiophene, 2-substituted thiophene, a C6-C12 fused ring, and an N-containing C6-C12 fused ring.

[0010] R 5 are independently selected from hydrogen, deuterium, halogen, a hydroxy group, a nitro group, a cyano group, a C1-C4 alkyl group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, and a C3-C6 cycloalkyl group.

[0011] R 6 are independently selected from hydrogen, deuterium, halogen, a hydroxy group, a nitro group, a cyano group, a C1-C4 alkyl group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, and a C3-C6 cycloalkyl group.

[0012] R 7 are independently selected from a substituted phenyl group, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, imidazole, pyrazole, oxazole, pyrazine, pyridazine, 2-furan, 2-substituted furan, thiophene, 2-substituted thiophene, substituted indole, a C6-C12 fused ring, and an N-containing C6-C12 fused ring.

[0013] The substituents in the 2-substituted furan, substituted phenyl and substituted indole are independently selected from a C1-C4 alkyl group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, a C1-C4 alkanol group, a cyano group, a halogen atom, a hydroxy group and an amine group.

[0014] Preferably, the R 1 are independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, and butyl; R 2are independently n-pentyl group, isopentyl group, n-butyl group, isobutyl group, 2-methoxyethyl group, 2,2-difluoroethyl group, 2-hydroxyethyl group, 1-hydroxypropyl group, 2-hydroxypropyl group, 3-hydroxypropyl group, 3-hydroxybutyl group, 2-pyridine, 3-pyridine, 4-pyridine, 2-furan, 2-substituted furan, thiophene, 2-substituted thiophene, substituted phenyl group, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, imidazole, pyrazole, oxazole, pyrazine, pyridazine, quinoline, isoquinoline, naphthalene, indole, substituted indole, [ka] Selected from R 4 are independently selected from substituted phenyl groups, 2-furan, 2-substituted furan, thiophene, 2-substituted thiophene, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, imidazole, pyrazole, oxazole, pyrazine, pyridazine, quinoline, isoquinoline, naphthalene, indole, and substituted indole; R 5 are independently selected from hydrogen, deuterium, halogen, hydroxy, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy, and cyclopropyl; R 6 are independently selected from hydrogen, deuterium, halogen, hydroxy, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy, and cyclopropyl; R 7 are independently selected from substituted phenyl groups, thiophene, 2-substituted thiophenes, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, imidazole, pyrazole, oxazole, pyrazine, pyridazine, 2-furan, 2-substituted furans, quinoline, isoquinoline, naphthalene, indole, and substituted indole; The substituents in the 2-substituted furan, substituted phenyl group, and substituted indole are a C1-C4 alkyl group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, a C1-C4 alkanol group, a cyano group, a halogen atom, a hydroxy group, and an amine group.

[0015] In one preferred embodiment of the present invention, the 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound is any one selected from the following structural formulas: [ka] TIFF0007784765000005.tif33170

[0016] The second object of the present invention is achieved by the following technical measures.

[0017] The method for producing the 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound includes the following steps: S1. According to the following reaction scheme, the compound represented by formula 1 is dissolved in an acidic solvent, cooled to 0-10°C, and then the compound represented by formula 2 is added dropwise. The mixture is then cooled to room temperature and reacted to obtain the compound represented by formula 3; [ka] S2. According to the following reaction scheme, the compound represented by formula 3 is cooled to 0-10°C, and then a solution of ammonia in 1,4-dioxane is added and reacted at room temperature under sealed conditions to obtain the compound represented by formula 4; [ka] S3. According to the following reaction scheme, the compound represented by formula 4 is dissolved in a basic organic solvent, cooled to 0-20°C, and then the compound represented by formula 5 is added and reacted at the same temperature to obtain the compound represented by formula 6; [ka] S4. According to the following reaction scheme, the compound represented by formula 6 is mixed with an ammonium salt and an acid, and then heated to 80-100°C and reacted at the same temperature to obtain the compound represented by formula 7; [ka] S5. According to the following reaction scheme, the compound represented by formula 7 is dissolved in a basic organic solvent, cooled to 0°C, and then di-t-butyl dicarboxylate is added. The mixture is then cooled to room temperature and reacted to obtain the compound represented by formula 8; [ka] S6. According to the following reaction scheme, the compound represented by formula 8 is dissolved in an organic solvent, cooled to 0°C, and then a basic substance is added. The mixture is then cooled to room temperature and reacted to obtain the compound represented by formula 9; [ka] S7. According to the following reaction scheme, the compound represented by formula 9 is dissolved in an organic solvent, cooled to 0°C, and then an oxidizing agent is added. The temperature is then returned to room temperature and reacted to obtain the compound represented by formula 10; [ka] S8. According to the following reaction scheme, the compound represented by formula 10 is dissolved in an organic solvent, cooled to 0°C, and then an oxidizing agent is added and reacted at the same temperature to obtain the compound represented by formula 11; [ka] S9. According to the following reaction scheme, the compounds represented by formula 11 and formula 12 are dissolved in an organic solvent, cooled to 0°C, and then a condensing agent is added. The mixture is then cooled to room temperature and reacted to obtain a compound represented by formula 13; [ka] S10. According to the following reaction scheme, the compound represented by formula 13 is dissolved in an organic solvent, cooled to 0°C, and then an acidic substance is added. The mixture is then cooled to room temperature and reacted to obtain the target compound.

[0018] [ka] R in the compound of formula I obtained by the above production process 1 If further modification of the group is required, it can be modified by referring to the usual method for modifying groups. For example, R 2 but [ka] When R 4 is a substituted phenyl group, the substituent of which is a 4-hydroxy group, R 4 When a compound in which R is a 4-methoxyphenyl group is subjected to a normal reaction, 4 It is possible to prepare a compound in which is a 4-hydroxyphenyl group.

[0019] The compound of the present invention has a novel structure, and the chemical synthesis method is simple and the conditions are mild, resulting in high yield and purity of the product, making it suitable for large-scale industrial production and easy to popularize and apply.

[0020] Preferably, in step S1, the acidic solvent comprises acetic acid, propionic acid and DMF (N,N-dimethylformamide). Specifically, the acidic solvent is acetic acid.

[0021] Preferably, in step S1, the molar mass ratio of the compound represented by formula 1 to the compound represented by formula 2 is 1:1.

[0022] Preferably, in step S2, the molar mass ratio of the compound represented by formula 3 to ammonia is 1:3-4.

[0023] More preferably, in step S2, the molar mass ratio of the compound represented by formula 3 to ammonia is 1:4.

[0024] Preferably, in step S3, the basic substance added to the basic organic solvent includes triethylamine, N,N-diisopropylethylamine, potassium carbonate, or cesium carbonate, and the organic solvent includes acetonitrile, dichloromethane, DMF (N,N-dimethylformamide), DMSO (dimethyl sulfoxide), or NMP (N-methylpyrrolidone). Specifically, the basic substance is N,N-diisopropylethylamine (DIPEA), and the organic solvent is acetonitrile.

[0025] Preferably, in step S3, the molar mass ratio of the compound represented by formula 4 to the compound represented by formula 5 is 1:1.

[0026] Preferably, in step S4, the ammonium salt includes ammonium acetate, ammonium sulfate, or ammonium nitrate, and the acid includes acetic acid or glacial acetic acid. Specifically, the ammonium salt is ammonium acetate, and the acid is acetic acid.

[0027] Preferably, in step S4, the molar mass ratio of the compound represented by formula 6 to the ammonium salt is 1:6, and the reaction temperature is 95°C.

[0028] Preferably, in step S5, the basic substance added to the basic organic solvent includes triethylamine and N,N-diisopropylethylamine, and the organic solvent includes dichloromethane, tetrahydrofuran, and 1,2-dichloroethane. Specifically, the basic substance is triethylamine, and the organic solvent is tetrahydrofuran.

[0029] Preferably, in step S5, the molar mass ratio of the compound represented by formula 7 to di-t-butyl dicarboxylate is 1:2.

[0030] Preferably, in step S6, the basic substance includes potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, or potassium hydroxide, and the organic solvent includes methanol, ethanol, or acetonitrile. Specifically, the basic substance is potassium carbonate, and the organic solvent is methanol.

[0031] Preferably, in step S6, the molar mass ratio of the compound represented by formula 8 to the basic substance is 1:2.

[0032] Preferably, in step S7, the oxidizing agent comprises Dess-Martin periodinane and pyridinium chlorochromate. Specifically, the oxidizing agent is Dess-Martin periodinane.

[0033] More preferably, in step S7, the molar mass ratio of the compound represented by formula 9 to the oxidizing agent is 1:1.

[0034] Preferably, in step S8, the oxidizing agent is sodium hypochlorite, and the organic solvent includes tetrahydrofuran and 2-methyltetrahydrofuran. Specifically, the organic solvent is tetrahydrofuran.

[0035] Preferably, in step S8, the molar mass ratio of the compound represented by formula 10 to the oxidizing agent is 1:2.

[0036] Preferably, in step S9, the compound represented by formula 12 is any one selected from 2-aminomethylpyridine, n-pentylamine, isobutylamine, 2,2-difluoroethylamine, 2-methoxyethylamine, ethanolamine, isopropanolamine, DL-aminopropanol, 3-amino-1-propanol instead of 2-aminomethylpyridine, aniline, p-methylaniline, m-aminoanisole, p-chloroaniline, 2-aminopyridine, 3-aminopyridine, benzylamine, 3-chlorobenzylamine, 3-aminomethylpyridine, 4-aminomethylpyridine, 2-furanmethylamine, 1-(2-pyridine)ethylamine instead of 2-aminomethylpyridine, (quinolin-8-yl)methanamine, (quinolin-8-yl)ethanamine, 2-(pyridin-2-yl)-2-propylamine, 2-(3-chlorophenyl)ethylamine, and 2-(4-methoxyphenyl)ethylamine.

[0037] Preferably, in step S9, the condensing agent includes a mixture of DMAP (4-dimethylaminopyridine) and DCC (dicyclohexylcarbodiimide), a mixture of DMAP and EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), a mixture of HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) and DIPEA (N,N-diisopropylethylamine), a mixture of HOBt (1-hydroxybenzotriazole) and DCC, or a mixture of HOBt and EDCI. Specifically, the condensing agent is a mixture of HOBt and DCC, and the molar ratio of HOBt to DCC is 1:1.

[0038] Preferably, in step S9, the molar mass ratio of the compound represented by formula 11 to the compound represented by formula 12 is 1:1.

[0039] Preferably, in step S10, the acidic substance includes trifluoroacetic acid, formic acid, and concentrated hydrochloric acid. Specifically, the acidic substance is trifluoroacetic acid.

[0040] Preferably, in step S10, the molar mass ratio of the compound represented by formula 13 to the acidic substance is 1:10.

[0041] The third object of the present invention is achieved by the following technical measures.

[0042] Use of the above 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound in the manufacture of an adenosine receptor inhibitor or an antitumor drug.

[0043] Preferably, the adenosine receptor is adenosine A 2A receptor, and the antitumor drug is A 2A It is a tumor immunotherapy drug that targets receptors.

[0044] The 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound of the present invention is adenosine A 2AAmong them, compounds 14a to 14e, 14k, 14o, 14p, 14r to 14u, 14w, 14x and 14ac to 14ae have good inhibitory effects on adenosine A 2A The inhibitory rate for the adenosine A receptor was 70% or more, and compounds 14a, 14r, 14s, 14u, and 14ac in particular had an inhibitory rate of 80% or more. 2A It exhibits excellent inhibitory effects on adenosine A 2A Receptor inhibitor or A 2A It can be manufactured into a tumor immunotherapy drug that targets the receptor.

[0045] Specifically, the present invention provides an adenosine receptor inhibitor having, as a main active ingredient, a 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound according to any one of claims 1 to 3.

[0046] Specifically, the present invention provides an antitumor drug having, as a main active ingredient, a 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound according to any one of claims 1 to 3.

[0047] Compared with the existing technology, the beneficial effects of the present invention are as follows:

[0048] The present invention discloses 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compounds, which are structurally novel and are a kind of adenosine receptor inhibitors, and are capable of inhibiting adenosine A 2A It has excellent inhibitory activity against adenosine A receptors and good subtype selectivity. 2A It is used in immunotherapy as a receptor-targeted inhibitor and adenosine A 2ABy inhibiting the activity of adenosine A receptor, the immune system can release its function of eliminating tumor cells, thereby achieving the effect of treating tumors. At the same time, the compound of the present invention is easy to prepare, uses inexpensive compounds as raw materials, has mild reaction conditions, has high total yield and purity, and can be produced on a scale of several tens of grams, making it suitable for subsequent clinical use. 2A This will be useful in researching candidate receptor inhibitors. DETAILED DESCRIPTION OF THE INVENTION

[0049] Specific embodiments of the present invention will be further described below. Note that the description of these embodiments is intended to facilitate understanding of the present invention, but does not limit the present invention. In addition, the technical features of each embodiment of the present invention described below may be combined as long as they do not conflict with each other.

[0050] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods, and the test materials used in the following examples are all commercially available unless otherwise specified.

[0051] Example 1 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14a) The specific manufacturing process is as follows:

[0052] (1) Preparation of Compound Represented by Formula 3: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 1 (1,2-phenylenediamine, 3.2 g, 30 mmol) and 50 mL of acetic acid were placed in a reaction bottle at room temperature and cooled in an ice bath. Then, the starting material, i.e., the compound represented by formula 1 (methyl 2,2,2-trichloroacetimidate, 4 mL, 30 mmol), was slowly added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. When the reaction was complete as indicated by TLC, the reaction mixture was filtered, and the cake was washed with water (3 times, 25 mL each time), and finally dried under vacuum to obtain the compound represented by formula 3 (2-trichloromethyl-benzopyrimidine) (7.1 g, 93%).

[0053] (2) Preparation of Compound Represented by Formula 4: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 3 (2-trichloromethylbenzopyrimidine, 5.9 g, 25 mmol) was cooled to 0°C, and then a solution of ammonia in 1,4-dioxane (0.40 M, 125 mL) was added at 0°C. After sealing, the mixture was stirred at room temperature for 2 hours. After TLC showed that the reaction was complete, the mixture was concentrated under reduced pressure and applied to a column to obtain the compound represented by formula 4 (2-cyanobenzopyrimidine, 2.7 g, 76%).

[0054] (3) Preparation of Compound Represented by Formula 6: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, MeCN (40 mL), DIPEA (4.4 g, 34 mmol), and the compound represented by formula 4 (2-cyanobenzopyrimidine, 2.6 g, 17 mmol) were added to a reaction vessel and cooled to -15 °C. At the same temperature, the compound represented by formula 5 (3-bromo-2-oxopropyl acetate, 3.3 g, 17 mmol) was added and stirred at -15 °C for 3 hours. When the reaction was complete as indicated by TLC, the mixture was diluted with 150 mL of water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and applied to a column to obtain the compound represented by formula 6 (1-(2-(4-methoxyphenyl)-2-oxaethyl)-1-hydro-2-cyanobenzopyrimidine, 3.5 g, 80%). The spectral information of the product is as follows: 1 H NMR (400 MHz, chloroform-d) δ 7.89-7.86 (m, 1H), 7.50-7.40 (m, 2H), 7.30-7.28 (m, 1H), 5.28 (s, 2H), 4.81 (s, 2H), 2.22 (s, 3H).

[0055] (4) Preparation of Compound Represented by Formula 7: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 6 (3-(2-cyano-1H-benzo[d]imidazol-1-yl)-2-oxopropyl acetate, 3.3 g, 13 mmol), ammonium acetate (5.1 g, 66 mmol), and 5 mL of acetic acid were placed in a sealed tube and stirred at 95 °C for 1 hour. After TLC showed the reaction was complete, saturated sodium bicarbonate solution was added to neutralize the reaction until no more bubbles were generated. The mixture was then extracted with dichloromethane (three times, each time with 100 mL of dichloromethane). The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and applied to a column to obtain the compound represented by formula 7 (1-aminobenzo[4,5]imidazo[1,2-a]pyrazin-3-yl)methyl acetate, 2.8 g, 85%). The spectral information of the product is as follows: 1H NMR (500 Mhz, chloroform-d) δ 7.96-7.86 (m, 3H), 7.56-7.46 (m, 3H), 6.06 (s, 2H), 5.10 (s, 2H), 2.16 (s, 3H).

[0056] (5) Preparation of Compound Represented by Formula 8: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 7 (1-aminobenzo[4,5]imidazo[1,2-a]pyrazin-3-yl)methyl acetate, 2.8 g, 11 mmol), DMAP (134 mg, 1.1 mmol), and triethylamine (4.5 g, 44 mmol) were dissolved in 50 mL of tetrahydrofuran and cooled to 0°C. Then, di-t-butyl dicarbonate (6.0 g, 27.5 mmol) was slowly added dropwise at 0°C, and the mixture was stirred at room temperature for 4 hours. After stirring, the reaction mixture was shown to be complete by TLC. Then, 150 mL of water was added and extracted with dichloromethane (three times, each extraction with 100 mL of dichloromethane). The organic phases were combined, washed with 100 mL of saturated brine, dried over sodium sulfate, concentrated under reduced pressure, and applied to a column to obtain the compound represented by formula 8 ((1-(di(t-butoxycarbonyl)amino)benzo[4,5]imidazo[1,2-a]pyrazin-3-yl)methyl acetate, 4.2 g, 84%). The spectral information of the product is as follows: 1 H NMR (400 MHz, chloroform-d): 8.46 (s, 1 H), 8.07 (d, J=8.3 Hz, 1 H), 7.97 (d, J=8.3 Hz, 1 H), 7.64 (t, J=7.7 Hz, 1 H), 7.53 (t, J=7.7 Hz, 1 H), 5.29 (s, 2 H), 2.16 (3 H, s), 1.41 (s, 18 H).

[0057] (6) Preparation of Compound Represented by Formula 9: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 8 (methyl 1-(di(t-butoxycarbonyl)aminobenzo[4,5]imidazo[1,2-a]pyrazin-3-yl)acetate, 4.1 g, 9 mmol) was dissolved in 50 mL of methanol, cooled to 0 ° C, and then potassium carbonate (3.7 g, 27 mmol) was slowly added at 0 ° C. The mixture was stirred at room temperature for 4 hours. When the reaction was complete as determined by TLC, 200 mL of water was added and the mixture was extracted with dichloromethane (three times, each time with 100 mL of dichloromethane). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then column-filtered to obtain the compound represented by formula 9 (t-butyl (3-(hydroxymethyl)benzo[4,5]imidazo[1,2-a]pyrazin-3-yl)carbamate, 67%, 1.9 g). The spectral information of the product is as follows: 1 H NMR (400 MHz, chloroform-d) δ 8.56 (s, 1H), 8.12 (s, 1H), 7.96 (d, J=8.3 Hz, 1H), 7.89 (d, J=8.3 Hz, 1H), 7.61-7.57 (m, 1H), 7.50-7.60 (m, 1H), 4.85 (s, 2H), 1.56 (s, 9H).

[0058] (7) Preparation of Compound Represented by Formula 10: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 9 ((3-(hydroxymethyl)benzo[4,5]imidazo[1,2-a]pyrazin-1-yl) t-butyl carbamate, 1.9 g, 6 mmol) was dissolved in 50 mL of dichloromethane and cooled to 0 ° C. Dess-Martin periodinane (3.6 g, 8.4 mmol) was slowly added at 0 ° C. and stirred at room temperature for 4 hours. After TLC showed the reaction was complete, the mixture was filtered, and saturated sodium bicarbonate solution was added until no more bubbles were generated. Extraction was performed with dichloromethane (3 times, 100 mL dichloromethane each time). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then column-filtered to obtain the compound represented by formula 10 ((3-formylbenzo[4,5]imidazo[1,2-a]pyrazin-3-yl) t-butyl aminoformate, 1.5 g, 81%). The spectral information of the product is as follows: 1 H NMR (500 MHz, chloroform-d) δ 10.17 (s, 1H), 8.82 (s, 1H), 8.03–8.00 (m, 2H), 7.70–7.67 (m, 1H), 7.61–7.57 (m, 1H), 1.61 (s, 9H).

[0059] (8) Preparation of Compound Represented by Formula 11: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 10 ((3-formylbenzo[4,5]imidazo[1,2-a]pyrazin-3-yl)aminoformate t-butyl ester, 1.4 g, 4.5 mmol), sodium dihydrogen phosphate (2.2 g, 18 mmol), and isopentene (1.6 g, 22 mmol) were dissolved in a mixture of 40 mL of tetrahydrofuran and 20 mL of water. The mixture was cooled to 0°C, and then sodium chlorite (1.6 g, 18 mmol) was slowly added at 0°C. The mixture was stirred at 0°C for 4 hours, and TLC showed that the reaction was complete. After the reaction was completed, the reaction was quenched with aqueous sodium thiosulfate (3.6 g, 22 mmol) at 0°C and extracted with dichloromethane (three times, 100 mL dichloromethane each time). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting solid was washed with n-hexane to give the compound represented by formula 11 (1-((t-butoxycarbonyl)amino)benzo[4,5]imidazo[1,2-a]pyrazine-3-carboxylic acid, 886 mg, 60%). The spectral information of the product is as follows: 1 H NMR (400 MHz, DMSO-d6) δ9.46 (s, 1H), 8.56 (s, 1H), 7.98-7.96 (m, 1H), 7.65-7.54 (m, 2H), 1.50 (s, 9H).

[0060] (9) Preparation of Compound Represented by Formula 13: The chemical reaction formula for the manufacturing process is as follows: [ka] According to the above reaction scheme, the compound represented by formula 11 (1-((t-butoxycarbonyl)amino)benzo[4,5]imidazo[1,2-a]pyrazine-3-carboxylic acid, 67 mg, 0.2 mmol), the compound represented by formula 12 (2-aminomethylpyridine, 32 mg, 0.3 mmol), and HOBt (41 mg, 0.3 mmol) were dissolved in 2 mL of dichloromethane and cooled to 0°C. Then, DCC (62 mg) was added slowly at 0°C. g, 0.3 mmol) was added and stirred at room temperature for 16 hours. When TLC showed the reaction was complete, it was filtered through silica gel and washed with a dichloromethane / methanol mixture (dichloromethane:methanol = 50:1). The filtrate was concentrated under reduced pressure to obtain the crude product of the compound represented by formula 13 ((3-((pyridin-2-ylmethyl)aminocarbonyl)benzo[4,5]imidazo[1,2-a]pyrazin-3-yl)carbamate t-butyl). The spectral information of the product is as follows: 1 H NMR (400 MHz, chloroform-d) δ 9.00 (s, 1H), 8.95 - 8.92 (m, 1H), 8.63 (d, J = 4.8 Hz, 1H), 8.49 (s, 1H), 8.00 (d, J = 8.5 Hz, 2H), 7.70 - 7.62 (m, 2H), 7.58 - 7.54 (m, 1H), 7.39 (d, J = 7.8 Hz, 1H), 7.22 - 7.19 (m, 1H), 4.85 (d, J = 5.8 Hz, 2H), 1.60 (s, 9H).

[0061] (10) Preparation of target compounds: The chemical reaction formula for the manufacturing process is as follows: [ka] The crude product of the compound represented by formula 13 obtained in step (9) (t-butyl (3-((pyridin-2-ylmethyl)aminocarbonyl)benzo[4,5]imidazo[1,2-a]pyrazin-3-yl)carbamate) is dissolved in 2 mL of dichloromethane and cooled to 0°C. 0.2 mL of trifluoroacetic acid is slowly added at 0°C, and the mixture is stirred at room temperature for 4 hours. After TLC indicates the completion of the reaction, saturated aqueous sodium bicarbonate solution is added until no bubbles are generated, and the mixture is then cooled to 0°C. Extraction with methane (three times, 100 mL each time with dichloromethane) was performed. The organic phases were combined and washed with saturated brine (100 mL). The combined organic phases were then dried over anhydrous sodium sulfate, concentrated under reduced pressure, and applied to a column to obtain 45 mg of the target compound 14a (1-amino-N-(pyridin-2-ylmethyl)benzo[4,5]imidazo[1,2-a]pyrazine-3-formamide), with a yield of 70% (this yield is the combined yield of steps (9) and (10)). The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6)δ8.95 (s, 1H), 8.81 (t, J= 6.0 Hz, 1H), 8.56 (d, J= 4.8 Hz, 1H), 8.45 (d, J= 8.2 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.78 (t, J= 7.7 Hz, 1H), 7.56-7.46 (m, 4H), 7.39 (d, J= 7.8 Hz, 1H), 7.30 (t, J= 6.2 Hz, 1H), 4.68 (d, J= 5.8 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ164.2, 158.2 , 149.4, 143.6, 137.3, 136.3 , 130.6, 130.2, 126.4, 123.6, 122.8, 121.9, 120.7, 113.6, 111.5, 44.6.

[0062] Example 2 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14b) The process for producing compound 14b was the same as that for compound 14a in Example 1, except that in step S9, n-pentylamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14b (46 mg, 70%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, chloroform-d) δ 8.75 (s, 1H), 7.94 (dd, J= 17.1, 8.3 Hz, 2H), 7.77 (t, J= 5.3 Hz, 1H), 7.59-7.56 (m, 1H), 7.51-7.47 (m, 1H), 5.88 (s, 2H), 3.48 (q, J= 6.8 Hz, 2H), 1.64 (p, J= 7.3 Hz, 2H), 1.43-1.38 (m, 2H), 1.36-1.32 (m, 3H), 0.91-0.89 (m, 3H). 13 C NMR (126 MHz, Chloroform-d) δ 163.7, 148.1, 143.7, 135.7, 130.1, 129.7, 126.6, 123.8, 121.0, 111.7, 39.5, 31.5, 29.7, 26.7, 22.6, 14.0.

[0063] Example 3 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14c) The process for producing compound 14c was the same as that for compound 14a in Example 1, except that in step S9, isobutylamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14c (43 mg, 70%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 Mhz, Chloroform-d) δ 8.76 (s, 1H), 7.96 (dd, J= 16.6, 8.3 Hz, 2H), 7.84 (s, 1H), 7.59 (t, J= 7.7 Hz, 1H), 7.51 (t, J= 7.7 Hz, 1H), 5.77 (s, 2H), 3.33 (t, J= 6.6 Hz, 2H), 1.02 (d, J= 6.7 Hz, 6H). 13 C NMR (126 MHz, DMSO-d6) δ 163.9, 149.3, 143.6, 136.2, 130.6, 130.3, 126.4, 123.5, 120.6, 113.6, 111.2, 46.5, 28.8, 20.5.

[0064] Example 4 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14d) The process for producing compound 14d was the same as that for compound 14a in Example 1, except that in step S9, 2,2-difluoroethylamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 3 (44 mg, 75%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.46-8.44 (m, 1H), 8.41 (t, J= 6.2 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.58-7.55 (m, 1H), 7.52-7.47 (m, 3H), 6.32-6.01 (m, 1H), 3.80 (tt, J= 15.8, 4.2 Hz, 2H). 19 F NMR (471 MHz, DMSO-d6) δ-122.25.

[0065] Example 5 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14e) The process for producing compound 14e was the same as that for compound 14a in Example 1, except that in step S9, 2-methoxyethylamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14e (40 mg, 71%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 Mhz, Chloroform-d) δ 8.75 (s, 1H), 8.06 (s, 1H), 7.96 (dd, J= 17.6, 8.2 Hz, 2H), 7.59 (t, J= 7.5 Hz, 1H), 7.51 (t, J= 7.6 Hz, 1H), 5.79 (s, 2H), 3.69 (q, J= 5.2 Hz, 2H), 3.61 (t, J= 4.9 Hz, 2H), 3.43 (s, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 164.0, 149.4, 143.6, 136.2, 130.6 , 130.1, 126.4, 123.5, 120.6, 113.6, 111.3, 71.1, 58.4, 38.9.

[0066] Example 6 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14f) The process for preparing compound 14f was the same as that for compound 14a in Example 1, except that in step S9, ethanolamine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 5 (27 mg, 50%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.90 (s, 1H), 8.43 (d, J= 8.2 Hz, 1H), 8.23 ​​(t, J= 5.6 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.57-7.46 (m, 4H), 4.92 (t, J= 4.7 Hz, 1H), 3.56 (q, J= 4.8 Hz, 2H). 3.14 (t, J= 3.5 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.0, 149.4, 143.6, 136.2, 130.6, 130.2, 126.4, 123.6, 120.6, 113.6, 111.2, 60.3, 41.9.

[0067] Example 7 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14g) The process for producing compound 14g was the same as that for compound 14a in Example 1, except that in step S9, isopropanolamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14g (30 mg, 53%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (400 MHz, DMSO-d6) δ 8.91 (s, 1H), 8.44 (d, J= 8.1 Hz, 1H), 8.20-8.15 (m, 1H), 7.92 (d, J= 8.1 Hz, 1H), 7.58-7.55 (d, J= 12.1 Hz, 3H), 7.48 (t, J= 7.5 Hz, 1H), 4.94 (d, J= 4.2 Hz, 1H), 3.84-3.75 (m, 1H), 3.21-3.15 (M, 1H), 1.10 (d, J= 6.1 Hz, 3H).3 C NMR (126 MHz, DMSO-d6) δ 163.9, 149.4, 143.5, 136.2, 130.6, 130.2, 126.4, 123.5, 120.6, 113.6, 111.2, 65.6, 46.6, 21.6.

[0068] Example 8 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14h) The process for preparing compound 14h was the same as that for compound 14a in Example 1, except that in step S9, DL-aminopropanol was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 14h (28 mg, 51%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.89 (s, 1H), 8.43 (d, J= 8.2 Hz, 1H), 8.01 (d, J= 8.5 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.58-7.54 (m, 3H), 7.48 (t, J= 7.7 Hz, 1H), 4.97 (s, 1H), 4.07-4.04 (M, 1H), 3.50-3.47 (m, 2H), 1.18 (d, J= 6.6 Hz, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 163.3, 149.4, 143.6, 136.2, 130.6, 130.3, 126.4, 123.5, 120.6, 113.6, 111.2, 64.6, 46.8, 17.9.

[0069] Example 9 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14i) The synthesis of compound 14i was the same as that of compound 14a in Example 1, except that in step S9, 3-amino-1-propanol was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, resulting in the synthesis of compound 14i (32 mg, 55%), whose chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.90 (d, J= 1.6 Hz, 1H), 8.44 (d, J= 8.3 Hz, 1H), 8.22 (t, J= 6.0 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.56 (t, J = 6.8 Hz, 1H), 7.49-7.46 (m, 3H), 4.58 (t, J= 5.0 Hz, 1H), 3.51 (q, J= 5.3 Hz, 2H), 3.42 (q, J= 6.2, 5.6 Hz, 2H), 1.71 (p, J= 6.6 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.0, 149.3, 143.6, 136.2, 130.6, 130.4, 126.4, 123.5, 120.6, 113.6, 111.2, 59.1, 36.6, 32.9.

[0070] Example 10 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14j) The process for producing compound 14j was the same as that for compound 14a in Example 1, except that in step S9, aniline was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14j (47 mg, 78%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 10.09 (s, 1H), 9.08 (s, 1H), 8.49 (d, J= 8.2 Hz, 1H), 7.95 (d, J= 8.2 Hz, 1H), 7.81 (d, J= 7.9 Hz, 2H), 7.62-7.57 (m, 3H), 7.51 (t, J= 7.6 Hz, 1H), 7.41 (t, J= 7.5 Hz, 2H), 7.15 (t, J= 7.3 Hz, 1H). 13 C NMR (126 MHz, DMSO-d6) δ 162.5, 149.3, 143.7, 138.6, 136.2, 130.6, 130.0, 129.5, 126.5, 124.3, 123.7, 120.7, 120.0, 113.7, 112.1.

[0071] Example 11 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14k) The process for producing compound 14k was the same as that for compound 14a in Example 1, except that in step S9, p-methylaniline was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14k (48 mg, 76%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (400 MHz, DMSO-d6) δ 9.99 (s, 1H), 9.05 (s, 1H), 8.49-8.47 (m, 1H), 7.94 (d, J= 7.9 Hz, 1H), 7.68-7.50 (m, 6H), 7.22-7.18 (d, J= 7.6 Hz, 2H), 2.28 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ 162.3, 149.3, 143.7, 136.2, 136.1, 133.3, 130.6, 130.1, 129.8, 126.5, 123.7, 120.7, 120.0, 113.7, 112.0, 21.0.

[0072] Example 12 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14l) The process for producing compound 11 was the same as that for compound 14a in Example 1, except that in step S9, m-aminoanisole was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 11 (46 mg, 69%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 10.06 (s, 1H), 9.07 (s, 1H), 8.48 (d, J= 8.0 Hz, 1H), 7.94 (d, J= 8.3 Hz, 1H), 7.61-7.51 (m, 6H), 7.36-7.31 (m, 2H), 6.72 (d, J= 8.1 Hz, 1H), 3.79 (s, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 162.5, 160.2, 149.3, 143.7, 139.8, 136.2, 130.6, 130.3, 130.0, 126.6, 123.7, 120.7, 113.7, 112.3, 112.2, 109.8, 105.8, 55.55.

[0073] Example 13 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14m) The process for producing compound 14m was the same as that for compound 14a in Example 1, except that in step S9, p-chloroaniline was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14m (41 mg, 61%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 10.21 (s, 1H), 9.08 (s, 1H), 8.49 (d, J= 7.0 Hz, 1H), 7.95-7.85 (m, 3H), 7.60-7.45 (m, 6H). 13 C NMR (126 MHz, DMSO-d6) δ 162.7, 149.3, 143.7, 137.7, 136.2, 130.6, 129.9, 129.3, 127.9, 126.6, 123.7, 121.7, 120.7, 113.7, 112.3.

[0074] Example 14 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14n) The process for producing compound 14n was the same as that for compound 14a in Example 1, except that in step S9, 2-aminopyridine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14n (43 mg, 70%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 10.33 (s, 1H), 9.14 (s, 1H), 8.51 (d, J= 8.1 Hz, 1H), 8.39 (s, 1H), 8.33 (d, J= 8.3 Hz, 1H), 7.96-7.91 (m, 2H), 7.81 (s, 2H), 7.59 (t, J= 7.7 Hz, 1H), 7.51 (t, J= 7.8 Hz, 1H), 7.20 (s, 1H). 13 C NMR (126 MHz, DMSO-d6) δ 162.5, 151.3, 149.5, 148.9, 143.7, 139.1, 136.3, 130.7, 129.1, 126.6, 123.8, 120.7, 120.4, 113.7, 113.3, 112.5.

[0075] Example 15 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14o) The process for producing compound 14o was the same as that for compound 14a in Example 1, except that in step S9, 3-aminopyridine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14o (35 mg, 59%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 10.29 (s, 1H), 9.11 (s, 1H), 8.99 (s, 1H), 8.49 (d, J= 8.1 Hz, 1H), 8.35-8.27 (m, 2H), 7.95 (d, J= 8.2 Hz, 1H), 7.60-7.43 (m, 5H). 13C NMR (126 MHz, DMSO-d6) δ 163.3, 149.4, 145.2, 143.7, 142.1, 136.2, 135.5, 130.6, 129.8, 127.3, 126.6, 124.2, 123.7, 120.7, 113.7, 112.6.

[0076] Example 16 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14p) The process for producing compound 14p was the same as that for compound 14a in Example 1, except that in step S9, benzylamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14p (46 mg, 73%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.60 (s, 1H), 8.44 (d, J= 7.3 Hz, 1H), 7.92 (d, J= 7.6 Hz, 1H), 7.59-7.52 (m, 1H), 7.47 (s, 3H), 7.36 (s, 4H), 7.27 (s, 1H), 4.56 (d, J= 5.1 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.1, 149.4, 143.6, 139.8, 136.2, 130.6, 130.2, 128.9, 128.0, 127.5, 126.4, 123.5, 120.6, 113.6, 111.5, 42.8.

[0077] Example 17 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14q) The process for preparing compound 14q was the same as that for compound 14a in Example 1, except that in step S9, 3-chlorobenzylamine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 14q (44 mg, 62%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.77 (t, J= 6.5 Hz, 1H), 8.43 (d, J= 8.2 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.57 (t, J= 7.7 Hz, 1H), 7.50-7.45 (m, 3H), 7.41-7.37 (m, 2H), 7.33-7.32 (m, 2H), 4.55 (d, J= 6.3 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.3, 149.4, 143.6, 142.6, 136.2, 133.5, 130.8, 130.6, 130.1, 127.7, 127.3, 126.6, 126.5, 123.6, 120.7, 113.6, 111.6, 42.3.

[0078] Example 18 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14r) The process for producing compound 14r was the same as that for compound 14a in Example 1, except that in step S9, 3-aminopyridine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14r (50 mg, 79%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.74 (t, J= 5.9 Hz, 1H), 8.60 (s, 1H), 8.48 (d, J= 4.2 Hz, 1H), 8.45 (d, J= 8.2 Hz, 1H), 7.93 (d, J= 8.2 Hz, 1H), 7.77 (d, J= 7.6 Hz, 1H), 7.57 (t, J= 7.6 Hz, 1H), 7.52-7.42 (m, 3H), 7.41-7.35 (m, 1H), 4.59 (d, J= 6.1 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.4, 149.5, 149.4, 148.7, 143.6, 136.2, 135.8, 135.4, 130.6, 130.2, 126.4, 124.0, 123.6, 120.6, 113.6, 111.6, 40.5.

[0079] Example 19 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14s) The process for producing compound 14s was the same as that for compound 14a in Example 1, except that in step S9, 4-aminopyridine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14s (41 mg, 65%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.80 (t, J= 6.1 Hz, 1H), 8.52 (d, J= 5.0 Hz, 2H), 8.44 (d, J= 8.2 Hz, 1H), 7.93 (d, J= 8.2 Hz, 1H), 7.57 (t, J= 7.6 Hz, 1H), 7.50-7.45 (m, 3H), 7.33 (d, J= 4.9 Hz, 2H), 4.58 (d, J= 6.2 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.6, 150.0, 149.4, 148.9, 143.6, 136.3, 130.6, 130.1, 126.4, 123.6, 122.7, 120.7, 113.6, 111.7, 41.9.

[0080] Example 20 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14t) The process for producing compound 14t was the same as that for compound 14a in Example 1, except that in step S9, 2-furanmethylamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, and finally compound 14t (39 mg, 64%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 8.95 (d, J= 1.4 Hz, 1H), 8.46 (d, J= 8.2 Hz, 1H), 8.42 (t, J= 5.9 Hz, 1H), 7.93 (d, J= 8.2 Hz, 1H), 7.63-7.62 (m, 1H), 7.59-7.55 (m, 1H), 7.52-7.47 (m, 3H), 6.44-6.43 (m, 1H), 6.36-6.35 (m, 1H), 4.58 (d, J= 6.0 Hz, 2H). 13C NMR (126 MHz, DMSO-d6) δ 163.8, 152.4, 149.4, 143.6, 142.9, 136.2, 130.6, 130.0, 126.4, 123.6, 120.7, 113.6, 111.5, 111.0, 107.7, 36.1.

[0081] Example 21 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14u) The process for preparing compound 14u was the same as that for compound 14a in Example 1, except that in step S9, 1-(2-pyridine)ethylamine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, resulting in the final preparation of compound 14u (44 mg, 66%), whose chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, chloroform-d) δ 8.78-8.75 (m, 2H), 8.64 (d, J= 4.2 Hz, 1H), 7.98-7.92 (m, 2H), 7.68 (td, J= 7.6, 1.8 Hz, 1H), 7.58 (t, J= 7.7 Hz, 1H), 7.49 (t, J= 7.7 Hz, 1H), 7.35 (d, J= 7.8 Hz, 1H), 7.23-7.21 (m, 1H), 5.93 (s, 2H), 5.39 (p, J= 7.0 Hz, 1H), 1.64 (d, J= 6.9 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 163.2, 161.2, 149.4, 148.3, 143.6, 136.9, 135.8, 130.1, 129.9, 126.6, 123.8, 122.4, 121.5, 121.0, 111.7, 50.1, 22.4.

[0082] Example 22 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14v) The process for preparing compound 14v was the same as that for compound 14a in Example 1, except that in step S9, (quinolin-8-ylmethyl)amine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 14v (48 mg, 65%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1 H NMR (500 MHz, DMSO-d6) δ 9.06-9.05(m, 1H), 8.94 (s, 1H), 8.84 (t, J= 6.3 Hz, 1H), 8.45-8.42 (m, 2H), 7.93 (t, J= 9.1 Hz, 2H), 7.73 (d, J= 7.0 Hz, 1H), 7.64-7.54 (m, 3H), 7.49-7.46 (m, 3H), 5.16 (d, J= 6.3 Hz, 2H).

[0083] Example 23 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14w) The process for preparing compound 14w was the same as that for compound 14a in Example 1, except that in step S9, 1-(8-quinoline)ethylamine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, ultimately producing compound 14w (52 mg, 68%), whose chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 9.47 (d, J= 9.0 Hz, 1H), 9.14 (d, J= 4.3 Hz, 1H), 8.88 (s, 1H), 8.44-8.41 (m, 2H), 7.94-7.91 (m, 2H), 7.80 (d, J= 7.1 Hz, 1H), 7.64-7.45 (m, 6H), 6.05 (p, J= 7.1 Hz, 1H), 1.69 (d, J= 6.6 Hz, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 162.9 , 150.4, 149.4, 145.7, 143.6, 141.3, 137.3, 136.3, 130.6, 130.4, 128.9, 127.9, 127.3, 126.9, 126.4, 123.5, 121.9, 120.6, 113.6, 111.3, 47.4, 23.6.

[0084] Example 24 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14x) The synthesis of compound 14x was the same as that of compound 14a in Example 1, except that in step S9, 2-(pyridin-2-yl)-2-propylamine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, resulting in the synthesis of compound 14x (48 mg, 70%), whose chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (400 MHz, chloroform-d) δ 9.29 (s, 1H), 8.72 (s, 1H), 8.61 (d, J= 4.4 Hz, 1H), 7.96 (d, J= 8.3 Hz, 1H), 7.87 (d, J= 8.2 Hz, 1H), 7.72 (td, J= 7.9, 1.5 Hz, 1H), 7.57 (t, J= 7.6 Hz, 1H), 7.50-7.45 (m, 2H), 7.21-7.18 (m, 1H), 6.12 (s, 2H), 1.91 (s, 6H). 13 C NMR (101 MHz, Chloroform-d) δ 164.6, 163.1, 148.3, 148.1, 143.7, 136.9, 135.8, 130.7, 130.1, 126.5, 123.7, 121.8, 121.0, 119.4, 111.6, 111.4, 57.0, 27.8.

[0085] Example 25 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14y) The process for preparing compound 14y was the same as that for compound 14a in Example 1, except that in step S9, 2-(3-chlorophenyl)ethylamine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 14y (46 mg, 63%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 8.89 (s, 1H), 8.43 (d, J= 8.0 Hz, 1H), 8.25 (d, J= 6.5 Hz, 1H), 7.91 (d, J= 7.9 Hz, 1H), 7.56 (t, J= 7.2 Hz, 1H), 7.49-7.45 (m, 3H), 7.35-7.32 (m, 2H), 7.28-7.23 (m, 2H), 3.60 (q, J= 7.0 Hz, 2H), 2.89 (t, J= 6.4 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.0, 149.3, 143.6, 142.4, 136.2, 133.5, 130.7, 130.6, 130.2, 129.0, 127.9, 126.7, 126.4, 123.5, 120.6, 113.6, 111.3, 35.3.

[0086] Example 26 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14z) The process for preparing compound 14z was the same as that for compound 14a in Example 1, except that in step S9, 2-(4-methoxyphenyl)ethylamine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 14z (51 mg, 72%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 8.90 (s, 1H), 8.44 (d, J= 8.2 Hz, 1H), 8.19 (t, J= 6.1 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.56 (t, J= 7.6 Hz, 1H), 7.50-7.46 (m, 3H), 7.19 (d, J= 8.5 Hz, 2H), 6.88 (d, J= 8.5 Hz, 2H), 3.73 (s, 3H), 3.55 (q, J= 6.9 Hz, 2H), 2.81 (t, J= 7.4 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 163.9, 158.2, 149.3, 143.6, 136.2, 131.6, 130.6, 130.3, 130.1, 126.4, 123.5, 120.6, 114.3, 113.6, 111.2, 55.5, 40.9, 34.9.

[0087] Example 27 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14aa) The synthesis of the first part of compound 14aa was the same as that of compound 14z in Example 26. Then, compound 14z was used as an intermediate according to the following reaction scheme: the methoxy group was converted to a hydroxy group with boron tribromide. The specific steps were as follows: Compound 14z (36 mg, 0.1 mmol) was dissolved in 1 mL of dichloromethane, cooled to 0 ° C, and then a dichloromethane solution of boron tribromide (0.2 mmol, 0.2 mL) was added. The temperature was then returned to room temperature and the reaction was carried out. After purification by column chromatography, compound 14aa (31 mg, 88%) was produced. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 9.24 (s, 1H), 8.91 (s, 1H), 8.44 (d, J= 8.1 Hz, 1H), 8.18 (t, J= 5.4 Hz, 1H), 7.92 (d, J= 8.1 Hz, 1H), 7.56 (t, J= 7.5 Hz, 1H), 7.50-7.48 (m, 3H), 7.07 (d, J= 8.0 Hz, 2H), 6.72 (d, J= 8.0 Hz, 2H), 3.53 (q, J= 6.4 Hz, 2H), 2.76 (t, J= 7.0 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ163.9, 156.2, 149.3, 143.6, 136.2, 130.6, 130.3, 130.0, 129.8, 126.4, 123.5, 120.6, 115.7, 113.6, 111.2, 41.0, 35.0.

[0088] Example 28 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14ab) The process for preparing compound 14ab was the same as that for compound 14a in Example 1, except that in step S9, homopiperonylamine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 14ab (52 mg, 69%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 8.90 (s, 1H), 8.44 (d, J= 8.2 Hz, 1H), 8.17 (t, J= 6.1 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.56 (t, J= 7.6 Hz, 1H), 7.49-7.45 (m, 3H), 6.86-6.83 (m, 2H), 6.72 (d, J= 7.7 Hz, 1H), 5.98 (s, 2H), 3.55 (q, J= 6.9 Hz, 2H), 2.80 (t, J= 7.3 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 163.9, 149.3, 147.7, 146.0, 143.6, 136.2, 133.5, 130.6, 130.3, 126.4, 123.5, 122.0, 120.6, 113.6, 111.2, 109.4, 108.7, 101.2, 40.8, 35.5.

[0089] Example 29 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14ac) The process for preparing compound 14ac was the same as that for compound 14a in Example 1, except that in step S9, 2-(2-aminoethyl)pyridine was used instead of 2-aminomethylpyridine to prepare the compound represented by formula 13, and finally compound 14ac (48 mg, 71%) was prepared. Its chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 8.90 (s, 1H), 8.58 (d, J= 4.8 Hz, 1H), 8.45-8.42 (m, 2H), 7.92 (d, J= 8.1 Hz, 1H), 7.73 (t, J= 7.7 Hz, 1H), 7.56 (t, J= 7.7 Hz, 1H), 7.49-7.46 (m, 3H), 7.32 (d, J= 7.8 Hz, 1H), 7.27-7.24 (m, 1H), 3.74 (q, J= 6.7 Hz, 2H), 3.05 (t, J= 7.1 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 163.9, 159.6, 149.6, 149.3, 143.6, 137.1, 136.2, 130.6, 130.3, 126.4, 123.6, 123.5, 122.0, 120.6, 113.6, 111.2, 38.8, 37.6.

[0090] Example 30 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14ad) The synthesis of compound 14ad was the same as that of compound 14a in Example 1, except that in step S9, 3-(2-aminoethyl)pyridine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, resulting in the synthesis of compound 14ad (46 mg, 69%), whose chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 8.90 (s, 1H), 8.49 (s, 1H), 8.44-8.43 (m, 2H), 8.26 (t, J= 6.2 Hz, 1H), 7.92 (d, J= 8.3 Hz, 1H), 7.71-7.69 (m, 1H), 7.56 (t, J= 6.7 Hz, 1H), 7.49-7.45 (m, 3H), 7.35-7.32 (m, 1H), 3.62 (q, J= 6.8 Hz, 2H), 2.91 (t, J= 7.2 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.1, 150.3, 149.3, 148.0, 143.6, 136.7, 136.2, 135.3, 130.6, 130.2, 126.4, 124.0, 123.5, 120.6, 113.6, 111.3, 40.0, 32.9.

[0091] Example 31 Preparation of 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compound (compound 14ae) The synthesis of compound 14ae was the same as that of compound 14a in Example 1, except that in step S9, 4-(2-aminoethyl)pyridine was used instead of 2-aminomethylpyridine to produce the compound represented by formula 13, resulting in the synthesis of compound 14ae (39 mg, 59%), whose chemical structure is as follows: [ka] The spectral information of the product is as follows: 1H NMR (500 MHz, DMSO-d6) δ 8.89 (s, 1H), 8.49-8.48 (m, 2H), 8.43 (d, J= 8.3 Hz, 1H), 8.25 (t, J= 6.1 Hz, 1H), 7.92 (d, J= 8.2 Hz, 1H), 7.58-7.54 (m, 1H), 7.49-7.46 (m, 1H), 7.43 (s, 2H), 7.31-7.30 (m, 2H), 3.64 (q, J= 6.9 Hz, 2H), 2.92 (t, J= 7.2 Hz, 2H). 13 C NMR (126 MHz, DMSO-d6) δ 164.1, 150.0, 149.3, 148.8, 143.6, 136.2, 130.6, 130.2, 126.4, 124.7, 123.5, 120.6, 113.6, 111.3, 39.8, 34.9.

[0092] Example 32: 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide-based compounds adenosine A 2A Inhibitory effect on receptors 1-aminobenzo[4,5]imidazo[1,2-a]pyrazine-3-formamide compounds of Examples 1-31 and adenosine A 2A The receptor binding status was tested. The specific procedure was as follows: each tube was charged with membrane protein solution (60 μg / tube, containing 2 mg / mL of ADA) and radioactive ligand 1 nM [3H]-ZM241385; -4The mixture was mixed uniformly by shaking at different M concentrations and incubated at 37°C for 30 min. The reaction was stopped in a water bath, and the free and bound ligands were separated by vacuum filtration through GF / C glass fiber filters. The membranes were then washed three times with approximately 4 mL of pre-chilled 50 mM Tris-HCl. The membranes were then placed upside down (i.e., with the grid facing up) on a tray and heated for 3 min until dry. The dried small circular filter membranes were then placed in liquid scintillation tubes and 2 mL of scintillation fluid was added. [3H] counting was then performed in a multi-function liquid scintillation counter. Three duplicate tubes were set up for each binding point, and the average value was calculated. Finally, data analysis was performed to determine the adenosine A of the compounds. 2A The inhibition rate for the receptor was calculated. Compound inhibition rate (I%) = (total binding tube cpm - compound cpm) / total binding tube cpm x 100%

[0093] The measurement results are shown in Table 1. As can be seen from Table 1, compounds 14a-14ae inhibited the activity of adenosine A 2A Among them, compounds 14a to 14e, 14k, 14o, 14p, 14r to 14u, 14w, 14x and 14ac to 14ae have good inhibitory effects on adenosine A 2A The inhibitory rate for the adenosine A receptor was 70% or more, and compounds 14a, 14r, 14s, 14u, and 14ac in particular had an inhibitory rate of 80% or more. 2A It has shown excellent inhibitory effects on the adenosine A2A receptor. It is used in immunotherapy as an inhibitor targeting the adenosine A2A receptor, 2A By inhibiting the activity of the receptor, the immune system's ability to eliminate tumor cells is released, thereby achieving a tumor treatment effect. [Table 1]

[0094] Although the embodiments of the present invention have been described in detail above, the present invention is not limited to the described embodiments. Those skilled in the art may make various changes, modifications, substitutions and variations to these embodiments without departing from the principle and spirit of the present invention, and these changes, modifications and variations will still fall within the scope of protection of the present invention.

Claims

1. The structure is represented by formula (I): 【Transformation 56】 (However, R １ are independently selected from hydrogen and C1-C4 alkyl groups; R ２ are independently a C1-C6 alkyl group, a C1-C4 alkanol group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, a substituted phenyl group, 2-pyridine, 3-pyridine, 4-pyridine, 2-furan, a C4-C12 heterocycle, 【Chemistry 57】 Selected from The R ４ are independently selected from substituted phenyl groups, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, imidazole, pyrazole, oxazole, pyrazine, pyridazine, 2-furan, 2-substituted furan, thiophene, 2-substituted thiophene, substituted indole, C6-C12 fused ring, and N-containing C6-C12 fused ring; The R ５ are independently selected from hydrogen, deuterium, halogen, a hydroxy group, a nitro group, a cyano group, a C1-C4 alkyl group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, and a C3-C6 cycloalkyl group; The R ６ are independently selected from hydrogen, deuterium, halogen, a hydroxy group, a nitro group, a cyano group, a C1-C4 alkyl group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, and a C3-C6 cycloalkyl group; The R ７ are independently selected from substituted phenyl groups, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, imidazole, pyrazole, oxazole, pyrazine, pyridazine, 2-furan, 2-substituted furan, thiophene, 2-substituted thiophene, C6-C12 fused ring, and N-containing C6-C12 fused ring; The substituents in the 2-substituted furan, substituted phenyl group, and substituted indole are independently selected from a C1-C4 alkyl group, a C1-C4 haloalkyl group, a C1-C4 alkoxy group, a C1-C4 alkanol group, a cyano group, a halogen atom, a hydroxy group, and an amine group. A 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound having the following structural formula: 【Transformation 58】

2. Use of the 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound according to claim 1 in the manufacture of an adenosine receptor inhibitor or an antitumor drug.

3. The adenosine receptor is adenosine A ２Ａ receptor, and the antitumor drug is A ２Ａ The use according to claim 2, characterized in that it is a tumor immunotherapy drug that targets a receptor.

4. An adenosine receptor inhibitor, characterized in that the 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound according to claim 1 is a major active ingredient.

5. An antitumor drug comprising the 1-aminobenzo[4,5]imidazo[1,2]pyrazine-3-formamide compound according to claim 1 as a main active ingredient.

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