Preparation and use of 3-amino-pyrazine-2-carboxamide derivatives targeting fgfr

Abstract

The application belongs to the technical field of pharmaceutical chemistry, and discloses a preparation method and application of a 3-amino-pyrazine-2-amide derivative taking FGFR as a target point.The derivative has the following structure general formula (I) shown in the figure, wherein R1 and R2 substituents can be combined arbitrarily.The compound can effectively inhibit the phosphorylation of FGFR kinase, and can be used for the treatment of tumor diseases related to FGFR kinase mutations.In particular, the compound SN23 has good selective inhibitory effect on FGFR2 kinase, and has significant inhibitory effect on the proliferation of various tumor cells, and can be used for the research and development of antitumor drugs.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, and specifically relates to a method for preparing and using a 3-amino-pyrazine-2-amide derivative targeting FGFR. Background Technology

[0002] Fibroblast growth factor receptors (FGFRs) belong to the receptor tyrosine kinase superfamily. Their abnormally activated signaling pathways are closely related to the occurrence and development of tumors, making them important therapeutic targets for cancer. Currently, marketed drugs targeting FGFR include Erdafitinib (2019), Pemigatinib (2020), Infigratinib (2021), and Futibatinib (2022). Many small molecule FGFR inhibitors, including covalent and non-covalent inhibitors, are also under development and entering clinical trials. Most reported FGFR inhibitors are ATP-competitive inhibitors, and their binding "ATP pockets" are highly conserved, leading to poor selectivity, significant toxicity, and a high risk of drug resistance. For example, many marketed FGFR inhibitors are associated with hyperphosphatemia. Furthermore, the four FGFR subtypes (FGFR1-4) are structurally highly similar, but their mutation types and rates differ across different cancer types. Although the second-generation futibatinib has been approved for marketing, there is still a lack of FGFR inhibitors with good selectivity. Therefore, developing FGFR inhibitors with excellent specificity is of great significance and quite challenging, and is also one of the important directions in current FGFR inhibitor development. Summary of the Invention

[0003] To address the shortcomings of the aforementioned technologies, this invention provides a method for preparing and using a 3-amino-pyrazine-2-amide derivative targeting FGFR. This type of compound can effectively inhibit the phosphorylation of FGFR and can be used to treat various FGFR-related tumors.

[0004] To achieve the above-mentioned technical effects, the present invention is implemented through the following technical solution:

[0005] A 3-amino-pyrazine-2-amide derivative targeting FGFR, having the general structural formula shown in formula (I):

[0006]

[0007] R1 can be either methyl (-CH3) or hydrogen (-H).

[0008] The R2 group can be selected from any of the following substituents. The wavy line indicates the position of the substituent.

[0009]

[0010] The R1 and R2 groups can be combined arbitrarily.

[0011] Furthermore, the 3-amino-pyrazine-2-amide derivative-targeted FGFR small molecule inhibitor is one of compounds SN1-SN28, and R1 and R2 of SN1-SN28 are shown in the table below.

[0012]

[0013] This invention also provides a method for preparing the above-mentioned 3-amino-pyrazine-2-amide derivative targeting FGFR, comprising the following steps:

[0014] The synthetic steps for compounds SN1-SN6 are as follows:

[0015] (1) Synthesis of compound SN6: 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (500 mg, 1.42 mmol), (4-(hydroxymethyl)phenyl)boronic acid (260 mg, 1.71 mmol), and sodium carbonate (301 mg, 2.84 mmol) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (4 / 1, 8 mL, volume ratio) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (63 mg, 0.086 mmol) as a catalyst. The reaction mixture was heated at 90 °C for 10 h. After the reaction was completed, thin-layer chromatography (TLC) was used to monitor the reaction. The reaction was quenched with water (5 mL) and extracted with ethyl acetate (6 mL * 3 times). The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3) to give the target compound SN6, a light brown solid (459 mg, yield 85%).

[0016] (2) General method for the synthesis of compound SN1-4: In a dry 20 mL round-bottom flask, compound SN6 (118 mg, 0.38 mmol), catalytic amounts of 4-dimethylaminopyridine (4.68 mg, 0.038 mmol), triethylamine (0.106 mL, 0.76 mmol), and dry dichloromethane (4 mL) were added sequentially. The resulting mixture was stirred at room temperature for 15 minutes, and p-toluenesulfonyl chloride (62.96 mg, 0.57 mmol) was slowly added. The mixture was stirred at room temperature for another 5 minutes, followed by the addition of the corresponding aliphatic amine and cesium carbonate until the reaction was complete. The mixture was diluted with 5 mL of water and then extracted with ethyl acetate (6 mL * 3 times). Purification was performed by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) to obtain the target compound SN1-4.

[0017] (3) Synthesis of compound SN5: First, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (100 mg, 0.28 mmol), (4-(hydroxyethylpiperazine)phenyl)boronic acid (0.42 mmol), and sodium carbonate (60 mg, 0.56 mmol) were added sequentially to a 20 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (4 / 1 volume ratio, 4 mL in total) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes. Then, Pd(dppf)Cl2-CH2Cl2 (12 mg, 0.0817 mmol) was added as a catalyst. The reaction mixture was heated at 90 °C for 6 hours, and then tetrabutylammonium fluoride (2 mL) was added. The reaction was carried out at room temperature for 2 hours. After the reaction was completed, 5 mL of water was added to dilute the mixture, and then it was extracted with ethyl acetate (6 mL * 3 times). Purification by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) yielded the target compound SN5, a pale yellow solid (20 mg).

[0018] The general method for synthesizing compounds SN7-SN14: The synthesis of compounds SN7-SN14 involves two steps. The first step is the synthesis of the intermediate borate ester. Commercially available 4-ethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and a secondary fatty amine (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase was separated, and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain the crude borate ester intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 6–9 h. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3) to obtain the target compounds SN7–SN14 as solid powders, with yields of 10–85%.

[0019] The general method for synthesizing compounds SN15-SN-28: In a dry 50 mL round-bottom flask, 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one of synthesis steps SN7-SN14 (1.5 equivalents), and sodium carbonate (2.5 equivalents) were added sequentially. A mixed solvent of 1,4-dioxane and water (4 / 1 v / v) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalents) as a catalyst. The reaction mixture was heated at 100 °C for 6-9 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon-based protecting group, yielding the target compounds SN15-SN-28 as solid powders with yields of 22-99%.

[0020] This invention provides the use of a 3-amino-pyrazine-2-amide derivative targeting FGFR, wherein the use is in the preparation of an antitumor drug.

[0021] The 3-amino-pyrazine-2-amide derivative of the present invention exhibits certain anti-tumor cell proliferation activity. As a further preferred embodiment, the 3-amino-pyrazine-2-amide derivative is compound SN23.

[0022] Compared with existing technologies, the technical effects of the present invention are reflected in the following: the present invention provides a small molecule inhibitor targeting FGFR with a novel structure, and the compound SN23 exhibits excellent selectivity for FGFR2 and has a significant inhibitory effect on the proliferation of various tumor cells. Attached Figure Description

[0023] Figure 1 Flowchart of the preparation process for compounds SN1-SN28

[0024] Figure 2 Diagram showing the inhibitory activity of compounds SN1-SN28 against FGFR1-4.

[0025] Figure 3 Figure showing the inhibitory activity of compound SN23 on tumor cell proliferation. Detailed Implementation

[0026] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0027] This invention discloses a 3-amino-pyrazine-2-amide derivative targeting FGFR, which has the general structural formula shown in (I) below. A flowchart of the preparation process for this structure is attached to the specification. Figure 1 :

[0028]

[0029] The present invention employs the following general characterization methods for the following embodiments:

[0030] The product testing conditions in the following examples were as follows: Characterization of the examples was performed using 600Hz nuclear magnetic resonance spectroscopy (Bruker Avance III 600), with deuterated chloroform or deuterated dimethyl sulfoxide as solvents; tetramethylsilane (TMS) was used as an internal standard. The NMR data of the compounds in the examples are reported below: 1 H NMR chemical shift, in δ; peak shape: s = singlet, d = doublet, t = triplet, m = multiplet; coupling constant J (in Hz). 13The C NMR data were reported as chemical shifts; the compounds in the examples were further validated using liquid chromatography-mass spectrometry (LC-MS, instrument model Agilent 7000D, Agilent Technologies, Inc.).

[0031] Example 1

[0032] Compound SN1 in this embodiment: 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-((4-ethylpiperazin-1-yl)methyl)phenyl)pyrazin-2-amide

[0033] The specific structural formula of compound SN1 is as follows: The preparation method of compound SN1 is as follows:

[0034] In a dry 20 mL round-bottom flask, compound SN6 (118 mg, 0.38 mmol), catalytic amounts of 4-dimethylaminopyridine (4.68 mg, 0.038 mmol), triethylamine (0.106 mL, 0.76 mmol), and dry dichloromethane (4 mL) were added sequentially. The resulting mixture was stirred at room temperature for 15 minutes, and p-toluenesulfonyl chloride (62.96 mg, 0.57 mmol) was slowly added. The mixture was stirred at room temperature for another 5 minutes, followed by the addition of 1-ethylpiperazine and cesium carbonate (1.5 equivalents) until the reaction was complete. The mixture was diluted with 5 mL of water and then extracted with ethyl acetate (6 mL x 3 times). Purification by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) yielded the target compound SN1 as a pale yellow solid in 50% yield. Its structural characterization data are as follows: 1 H NMR (600MHz, CDCl3) δ = 9.87 (s, 1H), 8.65 (s, 1H), 7.83–7.81 (m, 2H), 7.46–7.45 (m, 2H), 6.94 (d, J = 2.2Hz, 2H) ,6.29(t,J=2.2Hz,1H),3.82(s,6H),3.58(s,2H),2.52(s,6H),2.43(q,J=7.2Hz,4H),1.09(t,J=7.2Hz,3H). 13 C NMR(150MHz, CDCl3)δ=164.12,161.29,154.13,144.79,140.45,139.25,139.12,134.97,129.95,1 25.67,124.58,98.28,97.00,62.77,55.58,53.18,52.92,52.42,12.06.LC-MS(ESI)calculatedfor C 26 H 33N6O3,[M+H] + =477.25; found:477.30.

[0035] Example 2

[0036] Compound SN2 in this embodiment: 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-((4-methylpiperazin-1-yl)methyl)

[0037] The specific structural formula of SN2 of the phenylpyrazine-2-amide compound is as follows: The preparation method of compound SN2 is as follows:

[0038] In a dry 20 mL round-bottom flask, compound SN6 (118 mg, 0.38 mmol), catalytic amounts of 4-dimethylaminopyridine (4.68 mg, 0.038 mmol), triethylamine (0.106 mL, 0.76 mmol), and dry dichloromethane (4 mL) were added sequentially. The resulting mixture was stirred at room temperature for 15 min, and p-toluenesulfonyl chloride (62.96 mg, 0.57 mmol) was slowly added. The mixture was stirred at room temperature for another 5 min, followed by the addition of 1-methylpiperazine and cesium carbonate (1.5 equivalents) until the reaction was complete. The mixture was diluted with 5 mL of water and then extracted with ethyl acetate (6 mL x 3 times). Purification by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) yielded the target compound SN2 as a pale yellow solid in 60% yield. Its structural characterization data are as follows: 1 HNMR (600MHz, CDCl3) δ = 9.87 (s, 1H), 8.65 (s, 1H), 7.83 (d, J = 8.2Hz, 2H), 7.46 (d, J = 8.0Hz, 2H), 6 .94(d,J=2.2Hz,2H),6.29(t,J=2.3Hz,1H),3.83(s,6H),3.59(s,2H),2.57(s,8H),2.35(s,3H). 13 CNMR(151MHz, CDCl3)δ=164.09,161.26,154.12,144.77,140.35,139.22,138.93,135.02,129 .91,125.68,124.55,98.25,96.94,62.61,55.56,55.09,52.85,45.88.LC-MS(ESI)calculated for C 25 H 31 N6O3[M+H] + =463.24; found:463.20.

[0039] Example 3

[0040] Compound SN3 in this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-(piperazin-1-ylmethyl)phenyl)pyrazine-2-carboxamide.

[0041] The specific structural formula of compound SN3 is as follows: The preparation method of compound SN3 is as follows:

[0042] In a dry 20 mL round-bottom flask, compound SN6 (118 mg, 0.38 mmol), catalytic amounts of 4-dimethylaminopyridine (4.68 mg, 0.038 mmol), triethylamine (0.106 mL, 0.76 mmol), and dry dichloromethane (4 mL) were added sequentially. The resulting mixture was stirred at room temperature for 15 min, and p-toluenesulfonyl chloride (62.96 mg, 0.57 mmol) was slowly added. The mixture was stirred at room temperature for another 5 min, followed by the addition of piperazine and cesium carbonate (1.5 equivalents) until the reaction was complete. The mixture was diluted with 5 mL of water and then extracted with ethyl acetate (6 mL x 3 times). Purification by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) yielded the target compound SN3 as a pale yellow solid in 48% yield. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ=10.28(s,1H),8.90(s,1H),8.16(d,J=8.3Hz,2H),7.66(s,2H),7.43(d,J=8.1Hz,2H),7.13(d,J =2.2Hz,2H),6.32(t,J=2.3Hz,1H),3.77(s,6H),3.59(s,2H),3.04(t,J=5.0Hz,4H),2.61(t,J=1.9Hz,1H),2.56(s,4H). 13 C NMR (150MHz, DMSO-d6)δ=164.59,160.46,154.18,144.73,139.60,138.51,137.38,134.78,1 29.38,125.72,123.75,99.06,96.21,55.28,48.93,45.27,42.75.LC-MS(ESI)calculatedfor C 24 H 29 N6O3[M+H] + =,449.22; found:449.20.

[0043] Example 4

[0044] Compound SN4 in this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-(morpholinomethyl)phenyl)pyrazine-2-carboxamide. The specific structural formula of compound SN4 is as follows: The preparation method of compound SN4 is as follows:

[0045] In a dry 20 mL round-bottom flask, compound SN6 (118 mg, 0.38 mmol), catalytic amounts of 4-dimethylaminopyridine (4.68 mg, 0.038 mmol), triethylamine (0.106 mL, 0.76 mmol), and dry dichloromethane (4 mL) were added sequentially. The resulting mixture was stirred at room temperature for 15 minutes, and p-toluenesulfonyl chloride (62.96 mg, 0.57 mmol) was slowly added. The mixture was stirred at room temperature for another 5 minutes, followed by the addition of morpholine and cesium carbonate (1.5 equivalents) until the reaction was complete. The mixture was diluted with 5 mL of water and then extracted with ethyl acetate (6 mL x 3 times). Purification by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) yielded the target compound SN4 as a pale yellow solid in 56% yield. Its structural characterization data are as follows: 1 H NMR (600MHz, CDCl3) δ = 9.85 (s, 1H), 8.64 (s, 1H), 7.82 (d, J = 7.9Hz, 2H), 7.46 (d, J = 7.9Hz, 2H), 6.94 (d, J = 2.2Hz,2H),6.28(t,J=2.2Hz,1H),3.82(s,6H),3.74(t,J=4.7Hz,4H),3.56(s,2H),2.49(t,J=4.7Hz,4H). 13 C NMR (150MHz, CDCl3) δ = 164.05, 161.25, 154.13, 144.71, 140.24, 139.21, 135.08, 129. 92,125.66,124.55,98.26,96.89,67.06,63.11,55.54,53.71.LC-MS(ESI)calculated for C 24 H 28 N5O4[M+H] + =450.20; found:450.20.

[0046] Example 5

[0047] Compound SN5 of this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-(4-(2-hydroxyethyl)piperazin-1-yl)methyl)phenyl)pyrazine-2-carboxamide.

[0048] The specific structural formula of compound SN5 is as follows: The preparation method of compound SN5 is as follows:

[0049] In a 20 mL dry round-bottom flask, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (100 mg, 0.28 mmol), (4-(hydroxyethylpiperazine)phenyl)boric acid (0.42 mmol), and sodium carbonate (60 mg, 0.56 mmol) were added sequentially. A mixture of 1,4-dioxane and water (4 / 1 v / v, 4 mL) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min. Then, Pd(dppf)Cl2-CH2Cl2 (12 mg, 0.0817 mmol) was added as a catalyst. The reaction mixture was heated at 90 °C for 6 hours, followed by the addition of tetrabutylammonium fluoride (2 mL). The reaction was carried out at room temperature for 2 hours. After the reaction was complete, 5 mL of water was added to dilute the mixture, and then it was extracted with ethyl acetate (6 mL x 3 times). The solution was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) to give the target compound SN5 as a pale yellow solid. Its structural characterization data are as follows: 1 H NMR (600MHz, CDCl3) δ = 9.88 (s, 1H), 8.67 (s, 1H), 7.86–7.83 (m, 2H), 7.47 (d, J = 7.9Hz, 2H), 6.95 (d, J = 2.2Hz, 2H), 6 .30(t,J=2.2Hz,1H),3.84(s,6H),3.67(t,J=5.3Hz,2H),3.61(s,2H),3.35(d,J=8.7Hz,2H),2.64(t,J=5.2Hz,8H). 13 C NMR(150MHz, CDCl3)δ=164.13,161.34,154.18,144.81,140.41,139.25,135.21,130.00,125.80 ,124.69,98.36,97.03,62.52,59.57,59.13,57.60,55.63,53.00,52.56.LC-MS(ESI)calculated for C 26 H 33 N6O4[M+H] + =493.25; found:493.20.

[0050] Example 6

[0051] Compound SN6 in this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-(hydroxymethyl)phenyl)pyrazine-2-carboxamide. The specific structural formula of compound SN6 is as follows: The preparation method of compound SN6 is as follows:

[0052] In a dry 50 mL round-bottom flask, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (500 mg, 1.42 mmol), (4-(hydroxyethyl)phenyl)boric acid (260 mg, 1.71 mmol), and sodium carbonate (301 mg, 2.84 mmol) were added sequentially. A mixture of 1,4-dioxane and water (4 / 1 v / v, 8 mL) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (63 mg, 0.086 mmol) as a catalyst. The reaction mixture was heated at 90 °C for 10 h. After the reaction was complete as monitored by TLC, water (5 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (6 mL x 3 times). The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3) to give the target compound SN6, a light brown solid, in 85% yield. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ = 10.28 (s, 1H), 8.90 (s, 1H), 8.15 (d, J = 8.3Hz, 2H), 7.63 (brs, 2H), 7.43 ( d,J=8.3Hz,2H),7.13(d,J=2.2Hz,2H),6.32(t,J=2.2Hz,1H),4.56(d,J=5.7Hz,2H),3.77(s,6H). 13 C NMR(150MHz,DMSO-d6)δ=164.60,160.45,154.15,144.60,142.68,139.59,138.70,1 34.18,126.77,125.47,123.69,99.08,96.19,62.65,55.23.LC-MS(ESI)calculated for C 20 H 21 N4O4,[M+H] + =381.15; found:381.10.

[0053] Example 7

[0054] Compound SN7 in this embodiment: 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-(pyrrolidone-1-ylmethyl)phenyl)pyrazine-2-carboxamide

[0055] The specific structural formula of compound SN7 is as follows: The preparation method of compound SN7 is as follows:

[0056] To a 50 mL dry round-bottom flask, add commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent), acetonitrile (0.2 M), and triethylamine (0.65 mL, 2.0 equivalent) and tetrahydropyrrole (4.0 mmol, 2.0 equivalent) dropwise under stirring. The reaction mixture is stirred at room temperature for 4 hours. After the reaction is complete, the solvent is evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase is separated, and dried over anhydrous sodium sulfate. Concentrate under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which is used in the next step without further purification. In the second step, to a 50 mL dry round-bottom flask, add 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent). A mixture of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalents) as a catalyst. The reaction mixture was heated at 100°C for 6 hours. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3) to give the target compound SN7 as a pale yellow solid in 46% yield. Its structural characterization data are as follows: 46%. 1 H NMR (600MHz, CDCl3) δ=9.87(s,1H),8.66(s,1H),7.85–7.83(m,2H),7.49(d,J=8.0Hz,2H),6.95(d,J= 2.2Hz,2H),6.29(t,J=2.2Hz,1H),3.83(s,6H),3.72(s,2H),2.61–2.58(m,4H),1.83(p,J=3.1Hz,4H). 13 CNMR(150MHz, CDCl3)δ=164.15,161.31,154.14,144.84,140.49,139.26,135.00,129.79,1 25.77,124.61,98.30,97.06,77.16,60.30,55.60,54.22,23.59LC-MS(ESI)calculatedfor C 24 H 28 N5O3,[M+H] + =434.21; found:434.20.

[0057] Example 8

[0058] Compound SN8 in this embodiment: 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-((4-methylpiperidin-1-yl)methyl)

[0059] (Phenyl)pyrazine-2-amide.

[0060] The specific structural formula of compound SN8 is as follows: The preparation method of compound SN8 is as follows: Commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and 4-methylpiperidine (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase was separated, and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixture of 1,4-dioxane and water (4 / 1 v / v) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes. Then, Pd(dppf)Cl₂-CH₂Cl₂ (0.05 equivalents) was added as a catalyst, and the reaction mixture was heated at 100 °C for 6 h. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The target compound SN8 was obtained as a pale yellow solid with a yield of 16%. Its structural characterization data are as follows: 1 H NMR (600MHz, CDCl3) δ=9.87(s,1H),8.66(s,1H),7.84–7.82(m,2H),7.47(d,J=8.1Hz,2H),6.95(d,J=2.2Hz,2H),6.29(t,J=2.2Hz,1H),3.83(s,6 H),3.60(s,2H),2.93(dt,J=11.9,3.3Hz,2H),2.06–2.02(m,2H),1.65–1 .61(m,2H),1.42–1.34(m,2H),1.28–1.23(m,1H),0.93(d,J=6.2Hz,3H). 13C NMR(150MHz, CDCl3)δ=164.14,161.31,154.14,144.81,140.47,139.25,135.03,130.20,125.67 ,124.63,98.31,97.06,77.16,62.96,55.60,53.92,34.12,30.75,21.94.LC-MS(ESI)calculated for C 26 H 32 N5O3,[M+H] + =462.24; found:462.20.

[0061] Example 9

[0062] Compound SN9 in this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-((1,1-thiomorpholino)methyl)phenyl)pyrazine-2-carboxamide.

[0063] The specific structural formula of compound SN9 is as follows: The preparation method of compound SN9 is as follows: Commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and thiomorpholine-1,1-dioxide (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase was separated, and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixture of 1,4-dioxane and water (4 / 1 v / v) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalents) as a catalyst. The reaction mixture was heated at 100 °C for 6 h. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The target compound SN9 was obtained as a pale yellow solid in 10% yield. Its structural characterization data are as follows: 1HNMR (600MHz, CDCl3) δ = 9.85 (s, 1H), 8.66 (s, 1H), 7.86 (d, J = 7.8Hz, 2H), 7.46 (d, J = 7.8Hz, 2H), 6.95(d,J=2.2Hz,2H),6.30(t,J=2.2Hz,1H),3.83(s,6H),3.74(s,2H),3.07(d,J=30.6Hz,8H). 13 C NMR(150MHz, CDCl3)δ=164.11,161.46,154.35,144.85,140.18,139.29,135.85,129.6 4,126.10,124.82,ss98.57,97.07,61.37,55.65,51.75,50.87.LC-MS(ESI)calculated for C 24 H 28 N5O5S, [M+H] + =498.17; found:498.10.

[0064] Example 10

[0065] Compound SN10 in this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-((4-methyl-1,4-diazepine-1-yl)methyl)phenyl)pyrazine-2-amide.

[0066] The specific structural formula of compound SN10 is as follows: The preparation method of compound SN10 is as follows: Commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and N-methylperiperazine (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase was separated, and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixture of 1,4-dioxane and water (4 / 1, v / v) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalents) as a catalyst. The reaction mixture was heated at 100 °C for 6 h. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The target compound SN10 was obtained as a pale yellow solid in 54% yield. Its structural characterization data are as follows: 1 H NMR (600MHz, CDCl3) δ = 9.84 (s, 1H), 8.65 (s, 1H), 7.84–7.83 (m, 2H), 7.44 (d, J = 5.5Hz, 2H), 6.94 (d, J = 2.2Hz, 2H), 6.29 (d, J =2.2Hz,1H),3.82(s,6H),3.74(s,2H),3.34(d,J=5.1Hz,2H),3.17(s,2H),2.96(s,3H),2.85(s,2H),2.78(d,J=1.2Hz,4H). 13 C NMR (150MHz, CDCl3) δ = 164.02, 161.30, 154.22, 144.77, 139.98, 139.19, 129.73, 125.99, 125.96, 124. 64,98.37,96.86,66.25,62.15,55.60,54.96,53.66,49.04,44.52,23.82.LC-MS(ESI)calculatedfor C 26 H 33 N6O3,[M+H] + =477.25; found:477.20.

[0067] Example 11

[0068] Compound SN11 in this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-((4-(dimethylamino)piperidin-1-yl)methyl)phenyl)pyrazine-2-carboxamide).

[0069] The specific structural formula of compound SN11 is as follows: The preparation method of compound SN11 is as follows: Commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and 4-dimethylaminopiperidine (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase was separated, and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixture of 1,4-dioxane and water (4 / 1 v / v) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalents) as a catalyst. The reaction mixture was heated at 100 °C for 6 h. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The target compound SN11 was obtained as a pale yellow solid in 54% yield. Its structural characterization data are as follows: 1 H NMR (600MHz, CDCl3) δ = 9.87 (s, 1H), 8.65 (s, 1H), 7.83–7.81 (m, 2H), 7.44 (d, J = 8. 2Hz,2H),6.94(d,J=2.2Hz,2H),6.29(t,J=2.2Hz,1H),3.82(s,6H),3.55(s,2H), 2.98–2.95(m,2H),2.58(d,J=7.2Hz,4H),2.53(dt,J=11.6,3.7Hz,1H),2.02–1.9 8(m,2H),1.75–1.72(m,2H),1.60(dd,J=12.1,3.8Hz,2H),1.04(t,J=7.2Hz,6H). 13C NMR (150MHz, CDCl3) δ = 164.13, 161.28, 154.11, 144.79, 140.48, 139.57, 139.25, 134.84, 129.83, 125. 61,124.55,98.28,96.97,62.77,58.32,55.57,53.66,43.72,28.55,13.78.LC-MS(ESI)calculatedfor C 29 H 39 N6O3,[M+H] + =519.30; found:519.30.

[0070] Example 12

[0071] Compound SN12 in this embodiment is 3-amino-6-(4-((4-(diethylamino)piperidin-1-yl)methyl)phenyl)-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide.

[0072] The specific structural formula of compound SN12 is as follows: The preparation method of compound SN12 is as follows: Commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and 4-diethylaminopiperidine (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase was separated, and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixture of 1,4-dioxane and water (4 / 1 v / v) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalents) as a catalyst. The reaction mixture was heated at 100 °C for 6 h. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The target compound SN12 was obtained as a pale yellow solid in 53% yield. Its structural characterization data are as follows: 1H NMR(600MHz, CDCl3)δ=9.87(s,1H),8.65(s,1H),7.84–7.82(m,2H),7.44(d, J=8.1Hz,2H),6.95(d,J=2.2Hz,2H),6.29(t,J=2.2Hz,1H),3.83(s,6H),3.8 2(s,2H),3.57(s,2H),3.01(dd,J=9.5,6.4Hz,2H),2.61(td,J=8.1,4.0Hz,1 H), 2.45 (s, 6H), 2.06 (td, J = 11.9, 2.3Hz, 2H), 1.90 (dt, J = 12.9, 2.8Hz, 2H). 13 C NMR(150MHz, CDCl3)δ=164.06,161.24,154.12,144.69,140.26,139.20,138.87,135.00,129 .82,125.64,98.26,96.89,62.37,62.18,55.53,52.57,40.28,27.10.LC-MS(ESI)calculated for C 27 H 35 N6O3,[M+H] + =491.2726; found:491.2.

[0073] Example 13

[0074] Compound SN13 of this embodiment is 3-amino-N-(3,5-dimethoxyphenyl)-6-(4-((4-(4-(4-(pyrrolidin-1-yl)piperidin-1-yl)methyl)phenyl)pyrazine-2-carboxamide).

[0075] The specific structural formula of compound SN13 is as follows: The preparation of compound SN13 was as follows: Commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and 4-pyrrolidine-1-ylpiperidine (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), and the organic phase was separated and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate ester obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (4 / 1 volume ratio) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 8 h. After the reaction was completed as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The target compound SN13 was obtained as a pale yellow solid with a yield of 35%. Its structural characterization data are as follows: 1 HNMR (600MHz, CDCl3) δ = 9.88 (s, 1H), 8.66 (s, 1H), 7.84–7.81 (m, 2H), 7.47–7. 44(m,2H),6.95(d,J=2.3Hz,2H),6.29(t,J=2.2Hz,1H),3.83(s,6H),3.56(s,2 H),2.93(dt,J=12.3,3.2Hz,2H),2.70(d,J=6.2Hz,5H),2.04(td,J=11.8,2.3H z,2H),1.92–1.88(m,2H),1.83(p,J=3.2Hz,4H),1.69(qd,J=12.0,3.9Hz,2H). 13 C NMR(150MHz, CDCl3)δ=164.16,161.30,154.12,144.82,140.53,139.52,139.26,134.89,129.75,125.67, 124.59,98.31,97.04,77.16,62.51,62.10,55.60,52.51,51.23,30.70,23.42.LC-MS(ESI)calculatedfor C29 H 37 N6O3,[M+H] + =517.28; found:517.30.

[0076] Example 14

[0077] Compound SN14 in this embodiment: 6-(4-([1,4'-bispiperidine]-1'-ylmethyl)phenyl)-3-amino-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide.

[0078] The specific structural formula of compound SN14 is as follows: The preparation method of compound SN14 is as follows: Commercially available 4-bromomethylphenylboronic acid pinacol ester (2.0 mmol, 1.0 equivalent) and acetonitrile (0.2 M) solution were added to a 50 mL dry round-bottom flask. Triethylamine (0.65 mL, 2.0 equivalent) and 4-piperidinylpiperidine (4.0 mmol, 2.0 equivalent) were added dropwise under stirring. The reaction mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated, diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL, 3 times), the organic phase was separated, and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a crude borate intermediate as a grayish-white solid powder, which was used in the next step without further purification. In the second step, 3-amino-6-bromo-N-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide (1 equivalent), the borate obtained in step one (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixture of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalents) as a catalyst. The reaction mixture was heated at 100 °C for 8 h. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The target compound SN14 was obtained as a pale yellow solid with a yield of 41%. Its structural characterization data are as follows: 1H NMR(600MHz, CDCl3)δ=9.87(s,1H),8.65(s,1H),7.83–7.81(m,2H),7.45–7 .42(m,2H),6.94(d,J=2.2Hz,2H),6.29(t,J=2.2Hz,1H),3.82(s,6H),3.55( s,2H),3.01–2.98(m,2H),2.66(s,4H),2.02(td,J=11.9,2.2Hz,2H),1.90(d ,J=12.1Hz,2H),1.71(ddd,J=24.3,11.4,4.7Hz,7H),1.49(d,J=8.3Hz,2H). 13 C NMR(150MHz, CDCl3)δ=164.13,161.30,154.13,144.81,140.45,139.28,139.24,134.97,129.83,125.69 ,124.59,98.31,96.99,63.29,62.57,55.60,53.20,50.16,27.37,25.43,24.27.LC-MS(ESI),calculated for C 30 H 39 N6O3,[M+H] + =531.30; found:531.20.

[0079] Example 15

[0080] Compound SN15 in this embodiment is 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-(4-ethylpiperazin-1-yl)methyl)phenyl)pyrazine-2-carboxamide.

[0081] The specific structural formula of compound SN15 is as follows: The preparation method of compound SN15 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-(4-ethylpiperazine-1-yl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 6–9 h. The reaction was monitored by TLC until completion. The reaction was quenched with water and extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN15 as a pale yellow solid with a yield of 35%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ = 10.06 (s, 1H), 9.25 (s, 1H), 8.86 (s, 1H), 8.09 (d, J = 8.1Hz, 2H), 7.59 (s, 2H), 7.39 (d, J = 8.1Hz, 2H), 6. 77(d,J=2.2Hz,2H),6.04(t,J=2.2Hz,1H),3.50(s,2H),3.36(s,4H),2.39(s,4H),2.31(d,J=7.2Hz,2H),0.97(t,J=7.2Hz,3H). 13 C NMR (150MHz, DMSO-d6)δ=164.65,158.87,154.59,144.93,139.65,139.01,138.90,134.99,129 .66,125.95,124.38,99.52,99.29,62.23,53.07,52.85,52.06,12.43.LC-MS(ESI)calculated for C 24 H 29 N6O3,[M+H] + =449.22; found:449.20.

[0082] Example 16

[0083] Compound SN16 in this embodiment is 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-((4-methylpiperazin-1-yl)methyl)phenyl)pyrazine-2-carboxamide).

[0084] The specific structural formula of compound SN16 is as follows: The preparation method of compound SN16 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-((4-methylpiperazin-1-yl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0. Using 0.5 equivalents as a catalyst, the reaction mixture was heated at 100 °C for 8 h. After the reaction was complete, as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Tetrabutylammonium fluoride (2 mL) was then added to remove the silanyl protecting group, yielding the target compound SN16 as a pale yellow solid in 41% yield. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ=10.08(s,1H),9.32(s,2H),8.88(s,1H),8.12(d,J=8.1Hz,2H),7.63(s,2H),7.41(d,J= 8.0Hz,2H),6.76(d,J=2.1Hz,2H),6.04(t,J=2.2Hz,1H),3.57(d,J=7.2Hz,2H),3.51–3.49(m,8H),3.38(s,3H). 13 C NMR (150MHz, DMSO-d6)δ=164.18,158.44,154.16,144.49,139.16,138.49,134.71,129.3 4,125.55,123.96,99.10,98.92,61.32,53.92,51.32,44.49.LC-MS(ESI)calculatedfor C 23 H 27 N6O3,[M+H] + =435.21; found:435.20.

[0085] Example 17

[0086] Compound SN17 in this embodiment is 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-(piperazin-1-ylmethyl)phenyl)pyrazine-2-carboxamide.

[0087] The specific structural formula of compound SN17 is as follows:

[0088] The preparation method of compound SN17 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-(piperazine-1-ylmethyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 6 h. After the reaction was completed as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, and the resulting target compound SN17 was a light yellow solid powder with a yield of 31%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ = 10.12 (s, 1H), 9.94 (s, 2H), 8.95 (s, 1H), 8.29 (d, J = 8.1Hz, 2H), 7.79 ( d,J=8.0Hz,2H),6.76(d,J=2.1Hz,2H),6.06(t,J=2.2Hz,1H),3.56(s,2H),3.53–3.44(m,8H). 13 C NMR (150MHz, DMSO-d6) δ = 164.10, 158.42, 154.34, 144.61, 139.10, 137.63, 136.98, 132. 08,128.70,125.93,124.34,99.30,98.97,66.38,58.05,47.17.LC-MS(ESI)calculated for C 22 H 25 N6O3,[M+H] + =421.19; found:421.20.

[0089] Example 18

[0090] Compound SN18 in this embodiment is 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-(morpholinomethyl)phenyl)pyrazine-2-carboxamide.

[0091] The specific structural formula of compound SN18 is as follows:

[0092] The preparation method of compound SN18 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-(morpholinomethyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 7 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN18 as a pale yellow solid with a yield of 64%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ = 10.06 (s, 1H), 9.25 (s, 2H), 8.86 (s, 1H), 8.10 (d, J = 8.0Hz, 2H), 7.59 (brs, 2H), 7.41 (d ,J=8.0Hz,2H),6.77(d,J=2.2Hz,2H),6.04(s,1H),3.58(t,J=4.6Hz,4H),3.51(s,2H),2.38(t,J=4.6Hz,4H).. 13 C NMR (150MHz, DMSO-d6) δ = 164.21, 158.43, 154.16, 144.50, 139.22, 138.54, 137.97, 134. 65,129.36,125.54,123.96,99.10,98.86,66.25,62.14,53.20.LC-MS(ESI)calculated for C 22 H 24 N5O4, [M+H] + =422.17; found:422.10.

[0093] Example 19

[0094] Compound SN19 in this embodiment is 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-(4-(2-hydroxyethyl)piperazin-1-yl)methyl)phenyl)pyrazine-2-carboxamide.

[0095] The specific structural formula of compound SN19 is as follows:

[0096] The preparation method of compound SN19 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-(4-(2-hydroxyethyl)piperazine-1-yl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 7 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN19 as a pale yellow solid with a yield of 89%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ=10.08(s,1H),9.32(s,2H),8.87(s,1H),8.12(d,J=8.0Hz,2H),7.64(s,2H),7.41(d,J=7.9H z,2H),6.77(d,J=2.2Hz,2H),6.05(t,J=2.2Hz,1H),4.66(s,1H),3.61–3.53(m,4H),2.61(s,6H),2.57–2.47(m,4H). 13 C NMR (150MHz, DMSO-d6)δ=164.17,158.41,154.15,144.49,139.16,138.46,134.72,129.35 ,125.54,123.95,99.07,98.87,69.78,61.33,59.46,57.45,52.56.LC-MS(ESI)calculated for C 24 H 29 N₆O₄,[M+H] + =465.22; found:465.20.

[0097] Example 20

[0098] The specific structural formula of compound SN20 in this embodiment is as follows: 3-Amino-N-(3,5-dihydroxyphenyl)-6-(4-(hydroxymethyl)phenyl)pyrazine-2-carboxamide.

[0099] The preparation method of compound SN20 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-(hydroxymethyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes. Subsequently, Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) was added as a catalyst, and the reaction mixture was heated at 100 °C for 7 h. After the reaction was completed as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, and the resulting target compound SN20 was a pale yellow solid with a yield of 46%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6)δ=10.88(s,1H),10.07(s,1H),9.44(s,2H),8.88(s,1H),8.13(d,J =7.9Hz,2H),7.63(s,2H),7.42(d,J=7.9Hz,2H),6.79–6.74(m,2H),4.55(d,J=4.8Hz,2H). 13 C NMR (150MHz, DMSO-d6) δ = 164.1, 158.5, 154.1, 144.4, 142.8, 139.0, 138.6, 134.1, 126.8, 125.3, 123.8, 99.0, 57.5.LC-MS (ESI) m / z: calculated for C 18 H 17 N4O4,[M+H] + =353.12; found:353.10.

[0100] Example 21

[0101] Compound SN21 in this embodiment: 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-(pyrrolidone-1-ylmethyl)phenyl)pyrazine-2-carboxamide

[0102] The specific structural formula of compound SN21 is as follows:

[0103] The preparation method of compound SN21 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-(pyrrolidine-1-ylmethyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 7 h. After the reaction was completed as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN21 as a pale yellow solid with a yield of 92%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ = 10.10 (s, 1H), 9.31 (s, 2H), 8.89 (s, 1H), 8.15 (d, J = 8.0Hz, 2H), 7.70–7.61 (m, 2H) ),7.49(d,J=7.8Hz,2H),6.77(d,J=2.2Hz,2H),6.04(s,1H),3.85(s,2H),2.68(s,4H),1.80–1.76(m,4H). 13 C NMR (150MHz, DMSO-d6)δ=164.18,158.42,154.24,144.59,139.18,138.20,135.40,12 9.82,125.68,124.05,99.18,98.90,58.00,53.10,22.90.LC-MS(ESI)m / z:calculated for C 22 H 24 N5O3,[M+H] + =406.18; found:406.10.

[0104] Example 22

[0105] Compound SN22 in this embodiment: 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-((4-methylpiperidin-1-yl)methyl)phenyl)pyrazine-2-carboxamide

[0106] The specific structural formula of compound SN22 is as follows:

[0107] The preparation method of compound SN22 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-((4-methylpiperidin-1-yl)methyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 7 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN22 as a pale yellow solid with a yield of 22%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ = 9.29 (s, 2H), 8.87 (s, 1H), 8.10 (d, J = 8.2Hz, 2H), 7 .62(brs,2H),7.39(d,J=7.9Hz,2H),6.77(d,J=2.2Hz,2H),6.03(t,J=2.2Hz ,1H),3.50(s,2H),2.79(dt,J=11.6,3.5Hz,2H),1.97–1.91(m,2H),1.59–1 .55(m,2H),1.22(d,J=2.9Hz,1H),1.17–1.12(m,2H),0.89(d,J=6.5Hz,3H). 13 C NMR (150MHz, DMSO-d6)δ=164.21,158.41,154.14,144.50,139.20,138.56,131.55,129.31,128 .69,125.50,123.94,99.10,98.85,69.80,54.92,53.22,33.85,21.82.LC-MS(ESI)calculated for C 25 H 32 N5O3,[M+H] + =450.24; found:450.20.

[0108] Example 23

[0109] Compound SN23 in this embodiment is: 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-((1,1-dioxothiomorpholino)methyl)phenyl)pyrazine-2-carboxamide

[0110] The specific structural formula of compound SN23 is as follows:

[0111] The preparation method of compound SN23 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-((1,1-dioxothiomorpholino)methylphenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (…). Using 0.05 equivalents as a catalyst, the reaction mixture was heated at 100 °C for 7 h. After the reaction was complete, as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Tetrabutylammonium fluoride (2 mL) was then added to remove the silanyl protecting group, yielding the target compound SN23 as a pale yellow solid in 87% yield. Its structural characterization data are as follows: 1 HNMR (600MHz, DMSO-d6) δ = 10.10 (s, 1H), 9.30 (s, 2H), 8.88 (s, 1H), 8.13 (d, J = 8.0Hz, 2H), 7.64 (brs, 2H), 7.44 (d, J=8.0Hz,2H),6.77(d,J=2.1Hz,2H),6.03(t,J=2.2Hz,1H),3.73(s,2H),3.12(t,J=5.2Hz,4H),2.92–2.89(m,4H). 13 CNMR(150MHz,DMSO-d6)δ=164.20,158.40,154.16,144.53,139.21,138.44,137.71,134 .89,129.28,125.65,123.99,99.10,98.84,59.35,50.32,50.08.LC-MS(ESI)calculated for C 22 H 23 N5O5S, [M+H] + =470.14; found:470.10.

[0112] Example 24

[0113] Compound SN24 in this embodiment is: 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-(4-methyl-1,4-diaza-1-yl)methyl)phenyl)pyrazine-2-carboxamide

[0114] The specific structural formula of compound SN24 is as follows:

[0115] The preparation method of compound SN24 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-(4-methyl-1,4-diaza-1-yl)methyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 8 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN24 as a pale yellow solid with a yield of 33%. Its structural characterization data are as follows: 1 HNMR (600MHz, DMSO-d6) δ = 10.1 (s, 1H), 9.3 (s, 4H), 8.9 (s, 1H), 8.1 (d, J = 8.3Hz, 2H), 7.6 (brs, 2H), 7.4 (d, J = 8.0Hz, 2H), 6.8 (d, J = 2.2Hz, 3H), 6. 1(t,J=2.2Hz,1H),3.7(s,2H),2.8(t,J=5.7Hz,3H),2.8(d,J=5.7Hz,2H) ,2.7–2.7(m,3H),2.7(t,J=6.1Hz,3H),2.4(s,4H),1.8(q,J=5.8Hz,2H). 13 C NMR (150MHz, DMSO-d6)δ=164.21,158.44,154.14,144.49,139.19,138.51,134.54,129.00,128.74,12 5.55,123.94,99.07,98.87,61.40,56.89,55.39,53.60,45.36,25.61,13.52.LC-MS(ESI)calculated for C 24 H29 N6O3,[M+H] + =449.22; found:449.20.

[0116] Example 25

[0117] Compound SN25 in this embodiment: 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-((4-(dimethylamino)piperidin-1-yl)methyl)phenyl)pyrazine-2-carboxamide

[0118] The specific structural formula of compound SN25 is as follows:

[0119] The preparation method of compound SN25 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-((4-(dimethylamino)piperidin-1-yl)methyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 8 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, and the resulting target compound SN25 was a pale yellow solid with a yield of 94%. Its structural characterization data are as follows: 1 HNMR (600MHz, DMSO-d6) δ=10.08(s,1H),8.87(s,1H),8.10(d,J=8.2Hz,2H),7.63(brs,2H),7.38(d,J=8.1Hz,2H),6.77(d,J=2.1Hz,2H),6.03(t,J= 2.2Hz,1H),3.48(s,2H),2.85(d,J=11.5Hz,2H),2.20(s,6H),2.18(s,1H) ,2.13(tt,J=11.2,3.7Hz,2H),1.95–1.91(m,2H),1.72(d,J=11.8Hz,2H). 13C NMR (150MHz, DMSO-d6)δ=164.19,158.44,154.13,144.48,139.19,138.77,138.57,134.48,129 .14,125.49,123.92,99.06,98.86,61.75,61.70,52.32,41.31,27.91.LC-MS(ESI)calculated for C 25 H 31 N6O3,[M+H] + =463.24; found:463.20.

[0120] Example 26

[0121] Compound SN26 of this embodiment is 3-amino-6-(4-((4-(diethylamino)piperidin-1-yl)methyl)phenyl)-N-(3,5-dihydroxyphenyl)pyrazine-2-carboxamide.

[0122] The specific structural formula of SN26 is as follows:

[0123] The preparation method of compound SN26 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-((4-(diethylamino)piperidin-1-yl)methyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 8 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN26 as a pale yellow solid with a yield of 74%. Its structural characterization data are as follows: 1HNMR (600MHz, DMSO-d6) δ = 10.11 (s, 1H), 8.89 (s, 1H), 8.11 (d, J = 8.0Hz, 2H), 7. 67(s,2H),7.39(d,J=7.9Hz,2H),6.81(d,J=2.1Hz,2H),6.08(t,J=2.1Hz,1H),3 .49(s,2H),2.86(d,J=11.5Hz,2H),2.53–2.50(m,4H),2.48–2.44(m,1H),1.94 –1.91(m,2H),1.65(d,J=11.4Hz,2H),1.47–1.41(m,2H),0.96(t,J=7.1Hz,6H). 13 C NMR (150MHz, DMSO-d6)δ=164.18,158.46,154.15,144.47,139.19,138.74,138.59,134.47,129.15 ,125.46,123.91,99.05,98.89,61.77,57.68,52.76,42.97,28.12,13.49.LC-MS(ESI)calculated forC 27 H 34 N6O3,[M+H] + =491.27; found:491.20.

[0124] Example 27

[0125] Compound SN27 in this embodiment: 3-amino-N-(3,5-dihydroxyphenyl)-6-(4-((4-(pyrrolidin-1-yl)piperidin-1-yl)methyl)phenyl)pyrazin-2-carboxamide

[0126] The specific structural formula of compound SN27 is as follows:

[0127] The preparation method of compound SN27 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-((4-(pyrrolidone-1-yl)piperidin-1-yl)methyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 8 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN27 as a pale yellow solid with a yield of 54%. Its structural characterization data are as follows: 1 H NMR (600MHz, DMSO-d6) δ = 10.01 (s, 1H), 8.87 (s, 1H), 8.09 (d, J = 8.0Hz, 2H), 7.67 (b rs,2H),7.38(d,J=7.9Hz,2H),6.73(d,J=2.1Hz,2H),6.09(t,J=2.3Hz,1H),3.46( s,2H),3.25(d,J=7.2Hz,1H),2.77(d,J=11.1Hz,2H),2.50(t,J=1.9Hz,2H),2.44( d,J=6.1Hz,4H),1.94(d,J=10.6Hz,2H),1.79–1.75(m,2H),1.63(q,J=3.5Hz,4H). 13 C NMR (150MHz, DMSO-d6)δ=173.40,164.03,159.21,154.22,144.47,138.88,138.83,138.54,134.46,12 9.21,125.41,123.90,99.35,98.70,61.96,54.98,51.85,31.08,23.50,22.97.LC-MS(ESI)calculated for C 27 H 33 N6O3,[M+H] + =489.25; found:489.20.

[0128] Example 28

[0129] Compound SN28 in this embodiment: 6-(4-([1,4'-bispiperidin]-1'-ylmethyl)phenyl)-3-amino-N-(3,5-dihydroxyphenyl)pyrazine-2-carboxamide

[0130] The specific structural formula of compound SN28 is as follows:

[0131] The preparation method of compound SN28 is as follows: 3-amino-N-(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide (1 equivalent), 4-([1,4'-bispiperidine]-1'-ylmethyl)phenylboronic acid ester (1.5 equivalent), and sodium carbonate (2.5 equivalent) were added sequentially to a 50 mL dry round-bottom flask. A mixed solvent of 1,4-dioxane and water (volume ratio 4 / 1) was added to form a suspension. The suspension was degassed under nitrogen bubbling for 10 min, followed by the addition of Pd(dppf)Cl2-CH2Cl2 (0.05 equivalent) as a catalyst. The reaction mixture was heated at 100 °C for 8 h. After the reaction was complete as monitored by TLC, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3). Then, tetrabutylammonium fluoride (2 mL) was added to remove the silicon protecting group, yielding the target compound SN28 as a pale yellow solid with a yield of 54%. Its structural characterization data are as follows: 1 HNMR (600MHz, DMSO-d6) δ = 10.09 (s, 1H), 9.33 (s, 2H), 8.88 (s, 1H), 8.10 (d, J = 7.8 Hz,2H),7.65(brs,2H),7.37(d,J=7.9Hz,2H),6.78(d,J=2.2Hz,2H),6.05(d,J=2. 4Hz,1H),3.47(s,2H),2.85(d,J=11.1Hz,2H),2.26(s,1H),1.90(t,J=11.6Hz,2H ),1.69(d,J=12.0Hz,2H),1.47(ddt,J=16.4,12.0,4.2Hz,8H),1.38–1.35(m,2H). 13 C NMR (150MHz, DMSO-d6)δ=164.20,158.44,154.15,144.50,139.21,138.69,138.56,134.51,129.19,125 .48,123.91,99.06,98.85,62.13,61.76,52.65,49.64,27.33,25.62,14.11.LC-MS(ESI)calculatedfor C28 H 35 N6O3,[M+H] + =503.27; found:503.20.

[0132] Example 29

[0133] This embodiment tested the phosphorylation inhibition activity of compounds SN1-SN28 obtained in Examples 1-28 above. The detailed process is as follows: Step 1: Dilute 5× kinase buffer with ultrapure water to 1.33× kinase buffer. Dilute 200 ng / μL of FGFR1-4 protein to 0.6, 1.4, 0.5, and 4.0 ng / μL with 1.33× kinase buffer, and dilute the substrate peptide to 4 μM, labeled as 2× kinase / peptide mixture. Next, dilute 10 mM of ATP with 1.33× kinase buffer to 10, 20, 300, and 600 μM, labeled as 4× ATP. Finally, dilute the compound stock solution to 4× compound with ultrapure water, and perform the same dilution with DMSO. Dilute the phosphorylated substrate peptide (10 mM) to 4 μM with 1.33× kinase buffer, labeled as 2× P-peptide. Step 2 is the kinase reaction. First, dispense the solution into 384-well plates as follows, mix well, and incubate at room temperature in the dark for 1 hour. The solutions are as follows: Experimental wells: 5 μL 2× kinase / peptide mixture + 2.5 μL 4× compound + 2.5 μL 4× ATP; DMSO control wells: 5 μL 2× kinase / peptide mixture + 2.5 μL DMSO + 2.5 μL 4× ATP; Non-phosphorylated control wells: 5 μL 2× kinase / peptide mixture + 2.5 μL DMSO + 2.5 μL 1.33× kinase buffer; 100% phosphorylated control wells: 5 μL 2× P-peptide + 2.5 μL DMSO + 2.5 μL 1.33× kinase buffer. Step three is the Development reaction. Dilute the Development reagent with Development buffer to a 3% Development solution, add 5 μL to each well, mix well, and incubate at room temperature for 1 hour. Add 5 μL of stop solution to each well, mix well, and terminate the reaction. See the attached instruction manual for test results. Figure 2 .

[0134] Example 30

[0135] This embodiment tested the antitumor cell proliferation activity of compound SN23 obtained in Example 23 above. (1) Cells and culture medium: NCI-H520, SNU-16, KMS-11, SW-780 and BaF3 cells were cultured in RPMI1640 complete medium, and MDA-MB-453 cells were cultured in DMEM complete medium. The complete medium was prepared by adding 10% FBS (fetal bovine serum) and 1% penicillin-streptomycin antibiotics to the corresponding medium. (2) Cell proliferation test: Cell plating: Take cells from a 10cm dish, wash once with 4mL PBS, and add 1mL trypsin. Then, digest at room temperature for 1min, add 4mL of culture medium to stop digestion, and gently pipette to disperse the cells. Collect the cells into a 15mL centrifuge tube and centrifuge at 800rpm for 3min. After centrifugation, discard the supernatant, add 2ml of culture medium to resuspend, and mix the cell suspension by pipetting. Finally, take 10 μL of the resuspended cell suspension to fill the well, count the cells, and distribute them into 96-well plates according to the cell count, so that each well contains 190 μL of culture medium and 1×10⁻⁶ cells / well. 4 Cells were cultured in a 37°C CO2 (5%) incubator. The compound stock solution was removed from the -20°C freezer and diluted 50-fold with culture medium. Then, using a multi-channel pipette, the diluted compound was added to the corresponding wells (10 μL to 190 μL) to a 1000-fold dilution, with each well containing 0.1% DMSO. The 96-well plate was placed in a 37°C CO2 (5%) incubator for 72 h. After 72 h, the CellTiter-Glo reagent was removed from the -20°C freezer and thawed in a 37°C water bath. The 96-well plate was removed from the incubator, and 100 μL of supernatant from each well was aspirated using a multi-channel pipette. 40 μL of CellTiter-Glo reagent was added to each well using a multi-channel pipette, the container was capped, and the plate was placed on a shaker and incubated for 10 min at room temperature and in the dark. The chemiluminescence signal was then measured using a microplate reader with the parameters set. Data was collected and analyzed using GraphPad Prims 8 software. Results are attached to the instruction manual. Figure 3 .

Claims

1. A 3-amino-pyrazine-2-amide derivative targeting FGFR, characterized in that, The derivative has the general structural formula shown in equation (Ⅰ): Wherein, the R1 group is selected as either methyl or hydrogen; Wherein, the R2 group is selected from any of the following substituents, and the wavy line indicates the position of the substituent; The R1 and R2 groups can be combined arbitrarily.

2. A 3-amino-pyrazine-2-amide derivative targeting FGFR, characterized in that, It is one of the following compounds: 。 3. The method for preparing a 3-amino-pyrazine-2-amide derivative targeting FGFR according to claim 2, characterized in that... The following steps are included: The synthetic steps for compounds SN1-SN6 are as follows: Synthesis of compound SN6: 500 mg of 3-amino-6-bromo- ... N -(3,5-dimethoxyphenyl)pyrazine-2-carboxamide, 260 mg (4-(hydroxymethyl)phenyl)boric acid, and 301 mg sodium carbonate were added to a mixture of 1,4-dioxane and water to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of 63 mg Pd(dppf)Cl2-CH2Cl2 as a catalyst. The reaction mixture was heated to 90 °C. o Heating at C for 10 hours, the reaction was monitored by thin-layer chromatography (TLC) until completion. Then, 5 mL of water was added to quench the reaction, and the mixture was extracted with 6 mL of ethyl acetate. This extraction process was repeated three times. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain the target compound SN6. The general method for synthesizing compounds SN1-SN4: In a dry 20 mL round-bottom flask, 118 mg of compound SN6, 4.68 mg of catalytic amount of 4-dimethylaminopyridine, 0.106 mL of triethylamine, and 4 mL of dry dichloromethane were added sequentially. The resulting mixture was stirred at room temperature for 15 minutes, and 62.96 mg of p-toluenesulfonyl chloride was slowly added. The mixture was stirred at room temperature for another 5 minutes, followed by the addition of the corresponding aliphatic amine and cesium carbonate until the reaction was complete. The mixture was diluted with 5 mL of water, extracted three times with ethyl acetate, and purified by silica gel column chromatography to obtain the target compounds SN1-SN4, respectively. Synthesis of compound SN5: First, 100 mg of 3-amino-6-bromo- ... N 1,4-(3,5-dimethoxyphenyl)pyrazine-2-carboxamide, 0.42 mmol (4-(hydroxyethylpiperazine)phenyl)boric acid, and 60 mg sodium carbonate were added to a mixture of 1,4-dioxane and water to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of 12 mg Pd(dppf)Cl2-CH2Cl2 as a catalyst. The reaction mixture was heated to 90 °C. o After heating at C for 6 hours, 2 mL of tetrabutylammonium fluoride was added, and the reaction was carried out at room temperature for 2 hours. After the reaction was completed, 5 mL of water was added to dilute the mixture, and then it was extracted three times with ethyl acetate and purified by silica gel column chromatography to obtain the target compound SN5. The general method for synthesizing compounds SN7-SN14: The synthesis of compounds SN7-SN14 involves two steps. The first step is the synthesis of the intermediate borate ester. 2.0 mmol of commercially available 4-ethylphenylboronic acid pinacol ester and 0.2 M acetonitrile solution are added to a 50 mL dry round-bottom flask. Under stirring, 0.65 mL of triethylamine and 4.0 mmol of secondary fatty amine are added dropwise. The reaction mixture is stirred at room temperature for 4 hours. After the reaction is complete, the solvent is evaporated, diluted with 50 mL of ethyl acetate, and washed with 20 mL of saturated brine. This washing process is repeated three times. The organic phase is separated and dried over anhydrous sodium sulfate. The solution is concentrated under reduced pressure to obtain the crude borate ester intermediate as a grayish-white solid powder. The second step involves adding 1 equivalent of 3-amino-6-bromo- N -(3,5-dimethoxyphenyl)pyrazine-2-carboxamide, 1.5 equivalents of borate ester obtained in step one, and 2.5 equivalents of sodium carbonate were added to a mixed solvent of 1,4-dioxane and water to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of 0.05 equivalents of Pd(dppf)Cl2-CH2Cl2 as a catalyst. The reaction mixture was heated to 100 °C. o The reaction was heated at C for 6-9 hours. After the reaction was completed by TLC monitoring, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain the target compounds SN7-SN14 as solid powders. General method for synthesizing compounds SN15-SN28: In a 50 mL dry round-bottom flask, add 1 equivalent of 3-amino- N -(3,5-dihydroxyphenyl)-6-bromopyrazine-2-carboxamide, SN7-SN14 synthesis of step one yielded 1.5 equivalents of borate ester and 2.5 equivalents of sodium carbonate, which were then added to a mixed solvent of 1,4-dioxane and water to form a suspension. The suspension was degassed under nitrogen bubbling for 10 minutes, followed by the addition of 0.05 equivalents of Pd(dppf)Cl2-CH2Cl2 as a catalyst. The reaction mixture was then heated to 100... o The reaction was heated at C for 6-9 hours. After the reaction was completed by TLC monitoring, water was added to quench the reaction and the mixture was extracted three times with ethyl acetate. The resulting organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography. Then, 2 mL of tetrabutylammonium fluoride was added to remove the silicon-protecting group, and the target compounds SN15-SN28 were obtained as solid powders with a yield of 22-99%.

4. The method for preparing a 3-amino-pyrazine-2-amide derivative targeting FGFR according to claim 3, characterized in that: The compounds SN1-SN28 described in the steps were separated and purified by silica gel column chromatography, using petroleum ether / ethyl acetate in a volume ratio of 3:1 to 2:

1.

5. The method for preparing a 3-amino-pyrazine-2-amide derivative targeting FGFR according to claim 3, wherein the intermediate borate ester synthesized from compounds SN7-SN14 is characterized in that: No separation or purification is required.

6. The method for preparing a 3-amino-pyrazine-2-amide derivative targeting FGFR according to claim 3, wherein the mixed solvent of 1,4-dioxane and water is characterized in that: The volume ratio of 1,4-dioxane to water is 4 / 1.

7. The use of the 3-amino-pyrazine-2-amide derivative targeting FGFR according to claim 2 in the preparation of antitumor drugs.

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