A small molecule inhibitor of bromodomain-containing protein 4 of triazolopyridine class and preparation method and application thereof

Abstract

The application discloses a triazolopyridine bromodomain-containing protein 4 (BRD4) small molecule inhibitor and a preparation method and application thereof, a compound or a pharmaceutically acceptable salt or a racemate thereof, the general formula of which is shown as formula (I). The novel synthetic triazolopyridine compound effectively regulates the occurrence and development process of related diseases by inhibiting the activity of BRD4 protein. In addition to the advantage of inhibiting the activity of BRD4 protein, the compound has good water solubility and liver microsomal stability, which means that it can be better absorbed and distributed in the body, thereby improving the therapeutic effect. The compound also has good drugability, so that it can be better applied to various inflammatory, autoimmune diseases and tumor drugs. The application provides a new choice for the treatment of related diseases and is expected to play an important role in clinical practice.

Description

Technical Field

[0001] This invention relates to a triazolopyridine bromine domain protein 4 small molecule inhibitor, its preparation method and application, belonging to the field of medicinal chemistry. Background Technology

[0002] Bromodomain-containing protein 4 (BRD4) is a transcriptional and epigenetic regulator containing a bromodomain, belonging to the bromodomain and superterminal domain family. Its overexpression is closely related to the development of various cancers. BRD4 plays a crucial role in epigenetic regulation, recruiting positive transcription elongation factor b, activating RNA polymerase II, regulating DNA transcription, and controlling the expression of tumor growth factors and anti-apoptotic genes, such as the C-Myc gene, which is closely related to tumorigenesis and development. Epigenetic regulation refers to the regulation of gene expression phenotypes and functions without altering the DNA sequence, including post-translational modifications of histones such as histone acetylation, phosphorylation, and methylation, DNA methylation, and chromatin remodeling. Studies have demonstrated that epigenetic changes are closely related to tumorigenesis and development, and epigenetic regulation has become an important approach to cancer treatment. Among histone modifications, acetylated lysine residues at the histone tail are considered major epigenetic markers of chromatin opening and transcriptional activation. BRD4 acts as a "reader" for epigenetic regulation, utilizing its tandem BD domains to recognize acetylated lysine residues in histones, playing a crucial role in epigenetics. Therefore, inhibiting BRD4 expression may be a promising approach for cancer treatment.

[0003] Over the past decade, with the elucidation of the physiological functions of BRD4, significant progress has been made in the development of BRD4 inhibitors. Currently, more than ten BRD4 inhibitors have entered human clinical trials for the treatment of diseases such as cancer and inflammation. However, most of the BRD4 inhibitors in clinical trials have exhibited dose-limiting toxicity, thrombocytopenia, and gastrointestinal toxicity as side effects. Furthermore, studies have found that... BRD4 inhibitors have led to resistance, and no BRD4 inhibitors are currently on the market. Therefore, developing BRD4 inhibitors with novel scaffolds is significant for elucidating the physiological function of BRD4 and the mechanisms of resistance. Summary of the Invention

[0004] Purpose of the invention: The technical problem to be solved by the present invention is to provide a triazolopyridine BRD4 small molecule inhibitor that inhibits BRD4 protein activity, has good water solubility and liver microsomal stability, as well as its preparation method and application.

[0005] Technical solution: To solve the above-mentioned technical problems, the present invention provides a compound as shown in general formula (I) or a pharmaceutically acceptable salt or racemate thereof:

[0006]

[0007] Wherein, R1 is selected from groups with the following structures:

[0008]

[0009] R2 is selected from any mono- or di-substituted group at any position on the aromatic ring, and the substituent is selected from one or two of the following: halogen, CH3, OCH3, and CF3.

[0010] Ring A is selected from

[0011] R3 is selected from CH3, CH2CH3, or CH(CH3)2;

[0012] Y is selected from CH2 or NH.

[0013] This invention also provides a method for preparing the aforementioned compound or its pharmaceutically acceptable salt or racemate, wherein the synthetic route of the compound is as follows:

[0014]

[0015] Intermediates 3, 5, 7 or 3, 5, 10 are used to construct an imidazole ring under the action of Lewis acid Sc(OTf)3 to obtain the target compound J1-56.

[0016] The present invention also provides a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt or racemate thereof.

[0017] This also includes pharmaceutically acceptable carriers or excipients.

[0018] The present invention also provides the use of the said compound or its pharmaceutically acceptable salt, racemate or the said pharmaceutical composition in the preparation of an inhibitor having BRD4 inhibitory activity.

[0019] The present invention also provides the use of the said compound or a pharmaceutically acceptable salt, racemate or said pharmaceutical composition in the preparation of a medicament for treating inflammation, autoimmune diseases or tumors.

[0020] The concentration of the compound is 1–10 μM.

[0021] The present invention also provides the use of the aforementioned heterocyclic compounds or their pharmaceutically acceptable salts or racemates in the preparation of immune checkpoint inhibitors.

[0022] This invention includes the free form of compounds of formula (I), as well as pharmaceutically acceptable salts and stereoisomers thereof, wherein pharmaceutically acceptable salts include conventional non-toxic salts of compounds of this invention formed by reacting the compounds of this invention with inorganic or organic acids.

[0023] The use of the compound of formula (I) described in this invention, its stereoisomers, tautomers or pharmaceutically acceptable salts in the preparation of antitumor drugs.

[0024] As a preferred embodiment of the present invention, the compound of formula (I) is used as a medicament for various inflammatory, autoimmune diseases and tumors, including the small molecule inhibitor of triazolidine bromide domain protein 4 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0025] Dosage:

[0026] According to standard pharmaceutical techniques, the compounds of the present invention can be administered to mammals, preferably humans, alone or in combination with pharmaceutically acceptable receptors, excipients or diluents in a pharmaceutical composition; the compounds can be administered orally or subcutaneously, intramuscularly, intraperitoneally, intravenously, rectally and topically, through the eyes, lungs, nasal cavity or parenterally.

[0027] Drug metabolites and prodrugs:

[0028] Metabolites of the compounds and pharmaceutically acceptable salts involved in this application, as well as prodrugs that can be converted in vivo into structures of the compounds and pharmaceutically acceptable salts involved in this application, are also included in this application.

[0029] The purpose of this invention is to design small molecule compounds with the structural features of formula (Ⅰ). These compounds can effectively inhibit the activity of bromine domain protein 4 and have significant anti-proliferative activity against various cancer cell lines mediated by BRD4.

[0030] Beneficial Effects: Compared with existing technologies, this invention has the following significant advantages: 1. This novel synthetic triazolopyridine compound effectively regulates the occurrence and development of related diseases by inhibiting the activity of BRD4 protein; 2. In addition to the advantage of inhibiting BRD4 protein activity, these compounds also have good water solubility and liver microsomal stability, which means that they can be better absorbed and distributed in vivo, thereby improving the therapeutic effect; 3. These compounds also have good drug-like properties, making them more suitable for use in the treatment of various inflammatory diseases, autoimmune diseases, and tumors; 4. It provides new options for the treatment of related diseases and is expected to play an important role in clinical practice. Detailed Implementation

[0031] Example 1: Synthesis of N-(4-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-1)

[0032] Synthesis route:

[0033]

[0034] Step 1: Preparation of intermediate 5-bromo-2-hydrazine-1,2-dihydropyridine

[0035] Chemical structural formula:

[0036] The starting materials 2,5-dibromopyridine (3000 mg, 12.7 mmol), hydrazine hydrate (6.2 mL, 127 mmol), PEG 400 (12.7 mL), isobutanol (2.5 mL), and water (12.7 mL) were added to a round-bottom flask and reacted at 98 °C. After the reaction was completed by TLC monitoring, ice water (38 mL) was added at room temperature, and the mixture was stirred in an ice bath for 1 h. The mixture was filtered and dried under infrared light to give 1725.1 mg of a grayish-white product, with a yield of 72.45%. 1 H NMR (300MHz, Chloroform-d) δ8.13(s,1H),7.54(d,J=8.9Hz,1H),6.66(d,J=8.8Hz,1H),6.02(s,1H),3.53(s,2H).

[0037] Step 2: Preparation of intermediate 6-bromo-3-methyl-[1,2,4]triazolo[4,3-a]pyridine

[0038] Chemical structural formula:

[0039] The intermediate 5-bromo-2-hydrazide-1,2-dihydropyridine (1661.2 mg, 8.8 mmol) and acetyl chloride (15 mL) were added to a round-bottom flask and reacted at 60 °C. After the reaction was completed by TLC monitoring, n-hexane (10 mL) was added, and the mixture was stirred at room temperature for 15 min. The mixture was filtered and dried under infrared radiation. The crude product was dissolved in a two-phase mixture containing H₂O (15 mL) and dichloromethane (DCM) (15 mL), and the pH was adjusted to neutral. The mixture was stirred at room temperature for 15 min and extracted with DCM / H₂O. The organic layer was collected, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. 1567.9 mg of a grayish-white solid was obtained, with a yield of 83.69%. 1 H NMR (300MHz, Chloroform-d) δ8.07 (s, 1H), 7.68 (d, J = 9.6Hz, 1H), 7.34 (d, J = 1.7Hz, 1H), 2.79 (s, 3H).

[0040] Step 3: Preparation of intermediate 3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxaldehyde

[0041] Chemical structural formula:

[0042] Under anhydrous and oxygen-free conditions, the intermediate 6-bromo-3-methyl-[1,2,4]triazolo[4,3-a]pyridine (500 mg, 2.4 mmol) was dissolved in anhydrous THF (2.5 mL), and the mixture was cooled to -8 °C. i-PrMgCl (Annegi, W4200311000, 3.1 mL, 2.8 mmol) was dissolved in THF (2.8 mL, 2.8 mmol) and added dropwise to the reaction mixture over five minutes. The reaction mixture was then transferred to 0 °C and stirred for 30 minutes. DMF (NN-dimethylformamide, 0.9 mL, 5.6 mmol) was added, and the mixture was heated to 55 °C. After the reaction was complete as monitored by TLC, dichloromethane (5 mL) was added. The reaction mixture was slowly poured into a 10 wt% citric acid aqueous solution (6 mL), and the two phases were stirred. The aqueous layer was extracted with DCM; the organic layer was collected and dried over anhydrous sodium sulfate. Purified by silica gel column chromatography (DCM:MeOH = 60:1, v / v), 212.5 mg of a grayish-white solid was obtained, with a yield of 56.40%. 1 H NMR (300MHz, Chloroform-d) δ10.04(s,1H),8.51(d,J=1.3Hz,1H),7.83(d,J=9.1Hz,1H),7.75(d,J=8.2Hz,1H),2.90(s,3H).

[0043] Step 4: Preparation of intermediate N-(4-fluorobenzene)formamide

[0044] Chemical structural formula:

[0045] The starting materials 4-fluoroaniline (112 mg, 1 mmol), HCOONa (13.6 mg, 0.2 mmol), and HCOOH (2 mL) were added to a round-bottom flask and heated to 65 °C. The reaction was monitored by TLC until complete. Ethyl acetate (EA) (3 mL) and saturated NaHCO3 (3 mL) were added, and the mixture was stirred at room temperature for 15 min. The aqueous layer was extracted with EA, and the organic layer was collected and dried over anhydrous sodium sulfate. Purification was performed by silica gel column chromatography (PE:EA = 4:1, v / v) to give 172.3 mg of a grayish-white solid, with a yield of 86.13%.

[0046] Step 5: Preparation of intermediate 1-fluoro-4-isocyanuric benzene

[0047] Chemical structural formula:

[0048] Under anhydrous and anaerobic conditions, intermediate N-(4-fluorobenzamide) (100 mg, 0.5 mmol), anhydrous NET3 (anhydrous triethylamine, 0.7 mL, 5 mmol), and anhydrous THF (2 mL) were added to a two-necked flask and stirred at -5 °C for 10 min. Then, a solution of anhydrous THF (1 mL) containing POCl3 (0.12 mL, 1.25 mmol) was added dropwise over 35 minutes. After the reaction was complete as monitored by TLC, the reaction mixture was poured into a saturated K2CO3 solution at 0 °C. Extraction was performed by EA, and the solution was dried over anhydrous sodium sulfate. Alkaline alumina column chromatography (PE:EA = 40:1, v / v) yielded 70 mg of a white solid, with a yield of 76.50%.

[0049] Step 6: Preparation of intermediate (5-bromopyrazin-2-yl) tert-butyl carbamate

[0050] Chemical structural formula:

[0051] The starting material 2-amino-5-bromopyrazine (870.0 mg, 5.0 mmol) was dissolved in DCM (5 mL), and 4-DMAP (4-dimethylaminopyridine, 61.1 mg, 0.5 mmol) was added. At 0 °C, di-tert-butyl dicarbonate (1.7 mL, 7.5 mmol) was added dropwise. The reaction was carried out at room temperature, and the reaction was monitored by TLC until completion. The product was purified by silica gel column chromatography (PE:EA = 1:2, v / v) to give 799.5 mg of white solid, with a yield of 58.34%.

[0052] Step 7: Preparation of intermediate (5-morpholinopyrazin-2-yl) tert-butyl carbamate

[0053] Chemical structural formula:

[0054] (5-bromopyrazin-2-yl)carbamate tert-butyl ester (1000.0 mg, 3.6 mmol) and morpholine (6 mL) were added to a round-bottom flask and reacted at 75 °C. The reaction was monitored by TLC until completion, quenched with water, extracted with EA, and dried over anhydrous sodium sulfate. Purification by silica gel column chromatography (PE:EA = 5:1, v / v) yielded 413.5 mg of a white solid, with a yield of 40.98%. 1 H NMR (300MHz, CDCl3) δ8.75(s,1H),7.76(s,1H),7.13(s,1H),3.84(t,4H),3.45(t,4H),1.53(s,9H).

[0055] Step 8: Preparation of intermediate 5-morpholinpyrazine-2-amine

[0056] Chemical structural formula:

[0057] Intermediate (5-morpholinopyrazin-2-yl) tert-butyl carbamate (200 mg, 0.7 mmol), DCM (5 mL), and trifluoroacetic acid (2.5 mL) were added to a round-bottom flask and stirred at room temperature. The reaction was monitored by TLC until complete, quenched with saturated sodium bicarbonate solution, extracted with DCM, and dried over anhydrous sodium sulfate. Purification was performed by alkaline alumina column chromatography (PE:EA = 1:1, v / v) to give 112.7 mg of a yellow solid, with a yield of 87.60%.

[0058] Step 9: Preparation of N-(4-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-1)

[0059] Chemical structural formula:

[0060] Under anhydrous and oxygen-free conditions, Sc(OTf)3 (Bidet, BD31937, 40.4 mg, 0.08 mmol) was added to anhydrous DCM / MeOH (3 mL / 1 mL) solution containing the intermediate 3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxaldehyde (66.1 mg, 0.4 mmol) prepared in step 3 and the intermediate 5-morpholinopyrazine-2-amine (74.4 mg, 0.4 mmol) prepared in step 8. The mixture was stirred at room temperature for 30 min. Then, the intermediate 1-fluoro-4-isocyanate benzene (70.2 mg, 0.4 mmol) prepared in step 5 was added to the flask, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The reaction was quenched with saturated sodium bicarbonate solution, extracted with DCM, and purified by silica gel column chromatography (DCM:MeOH = 20:1, v / v) to give 34.5 mg of a yellowish-white solid, with a yield of 20.5%. 1 H NMR (300MHz, DMSO-d6) δ8.96(s,1H),8.62(d,J=17.0Hz,2H),7.95(d,J=9.7Hz,1H),7.79(d,J=9.7Hz,1H),7.38 -7.27(m,3H),6.55(d,J=8.3Hz,2H),3.75(d,J=4.6Hz,4H),3.27(d,J=4.6Hz,4H),2.64(s,3H).HRMS(ESI+)m / z calculated for C 23 H 21 FN8O[M+H] + ,445.1821;found,445.1821.

[0061] Example 2: Synthesis of J-10 to J-14

[0062] 1. Synthesis of N-(4-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-10)

[0063] Synthesis route:

[0064]

[0065] Steps 1 to 6 are the same as steps 1 to 6 in Example 1.

[0066] Step 7: Preparation of intermediate (5-(4-methylpiperazin-1-yl)pyrazin-2-yl)tert-butyl carbamate

[0067] Chemical structural formula:

[0068] The intermediate (5-bromopyrazin-2-yl)carbamate tert-butyl ester (1000.0 mg, 3.6 mmol) prepared in step 6 and N-methylpiperazine (6 mL) were added to a round-bottom flask and reacted at 85 °C. The reaction was monitored by TLC until completion, cooled to room temperature, quenched with water, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. Purification was performed by silica gel column chromatography (PE:EA = 1:4, v / v, 1% TEA) to give 313.5 mg of a white solid, with a yield of 34.98%. 1 H NMR (300MHz, CDCl3) δ8.75(s,1H),7.76(s,1H),7.13(s,1H),3.84(t,4H),3.45(t,4H),1.53(s,9H).

[0069] Step 8: Preparation of intermediate 5-(4-methylpiperazin-1-yl)pyrazin-2-amine

[0070] Chemical structural formula:

[0071] Intermediate (200 mg, 0.7 mmol) of 5-(4-methylpiperazin-1-yl)pyrazin-2-yl)carbamate tert-butyl ester, DCM (5 mL), and trifluoroacetic acid (2.5 mL) were added to a round-bottom flask and stirred at room temperature. The reaction was monitored by TLC until complete, quenched with saturated sodium bicarbonate solution, extracted with DCM, and dried over anhydrous sodium sulfate. Purification was performed by alkaline alumina column chromatography (PE:EA = 1:1, v / v) to give 178.7 mg of a yellow solid, with a yield of 91.60%.

[0072] Step 9: Preparation of 2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-fluorophenyl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-10)

[0073] Chemical structural formula:

[0074] Under anhydrous and anaerobic conditions, Sc(OTf)3 (Bide, BD31937, 40.4 mg, 0.08 mmol) was added to an anhydrous DCM / MeOH (3 mL / 1 mL) solution of intermediates methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxaldehyde (66.1 mg, 0.4 mmol) and 5-(4-methylpiperazin-1-yl)pyrazin-2-amine (69.4 mg, 0.4 mmol). The mixture was stirred at room temperature for 30 min. Then, intermediate 1-fluoro-4-isocyanuric benzene (70.2 mg, 0.4 mmol) was added to the flask, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The reaction was quenched with saturated sodium bicarbonate solution, extracted with DCM, and purified by silica gel column chromatography with DCM / MeOH (20 / 1 + 1% TEA) to give 42.5 mg of a yellowish-white solid, with a yield of 24.3%. 1 H NMR (300MHz, DMSO-d6) δ8.86(s,1H),8.54(s,1H),8.27(s,1H),7.96(d,J=9.8Hz,1H),7.79(d,J=9.7Hz,1H),7.41(s,1H), 7.21(d,J=8.3Hz,2H),6.61(d,J=8.3Hz,2H),3.17(s,4H)2.88(s,4H),2.64(s,3H)2.24(s,3H).HRMS(ESI+)m / zcalculated for C 24 H 24 FN9[M+H] + ,458.2139;found,458.2139.

[0075] 2. Synthesis of N-(4-chlorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-11)

[0076] Replacing 4-fluoroaniline with 1 mmol of 4-chloroaniline, the product was prepared according to the method described above, yielding 21.4 mg of a pale white solid with a yield of 16.7%. 1H NMR (300MHz, DMSO-d6) δ8.86(s,1H),8.54(s,1H),8.32(s,1H),8.08(d,J=9.0Hz,1H),7.50(s,1H),7.23( d,J=8.3Hz,2H),7.17(s,1H),6.61(d,J=8.3Hz,2H),3.78(s,4H)2.53(s,7H),2.22(s,3H).HRMS(ESI+)m / z calculated for C 24 H 24 ClN9[M+H] + ,474.1843;found,474.1843.

[0077] 3. Synthesis of N-(4-bromophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-12)

[0078] By replacing 4-fluoroaniline with 1 mmol of 4-bromoaniline, 19.2 mg of a pale yellow solid was prepared according to the above method, with a yield of 12.6%. 1 H NMR (300MHz, DMSO-d6) δ8.85(s,1H),8.45(s,1H),8.28(s,1H),8.14(d,J=9.9Hz,1H),7.75(d,J=9.7Hz,1H),7.5 2-7.46(m,2H),7.37-7.31(m,2H),7.09(s,1H),3.26(s,4H),2.43(d,J=6.7Hz,7H),2.22(s,3H).HRMS(ESI+)m / z calculated for C 24 H 24 BrN9[M+H] + ,518.1338;found,518.1338.

[0079] 4. Synthesis of N-(3-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-13)

[0080] By replacing 4-fluoroaniline with 1 mmol of 3-fluoroaniline, 29.2 mg of a yellow solid was prepared according to the above method, with a yield of 15.6%. 1H NMR (300MHz, DMSO-d6) δ8.86 (s, 1H), 8.37 (s, 1H), 8.14 (d, J = 9.9Hz, 1H), 7. 75(d,J=9.7Hz,1H),7.33(d,J=7.6Hz,1H),7.15(s,1H),7.09(d,J=1.9Hz,1H ),7.09(s,1H),6.89(d,J=7.7Hz,1H),6.79(s,1H),3.26(d,J=5.5Hz,4H),3 .60(d,J=5.6Hz,2H),2.73(s,2H),2.64(s,3H),2.22(s,3H).HRMS(ESI+)m / z calculated for C 24 H 24 FN9[M+H] + ,458.2139;found,458.2139.

[0081] 5. Synthesis of N-(2-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-14)

[0082] By replacing 4-fluoroaniline with 1 mmol of 2-fluoroaniline, 31.3 mg of a yellow solid was prepared according to the above method, with a yield of 21.3%. 1 H NMR (300MHz, DMSO-d6) δ8.82(s,1H),8.44(s,1H),8.27(s,1H),8.14(d,J=9.9Hz,1H),7.75(d,J=9.7Hz,1H),7.61 –7.54(m,1H),7.22–7.12(m,3H),7.09(s,1H),3.65(s,4H),2.46(s,4H),2.42(s,3H),2.23(s,3H).HRMS(ESI+)m / z calculated for C 24 H 24 FN9[M+H] + ,458.2139;found,458.2139.

[0083] Example 3: Synthesis of compounds J-20 to J-25

[0084] 1. Synthesis of 2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-fluorophenyl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-20)

[0085] Synthesis route:

[0086]

[0087] Step 1: Prepare intermediate 5-bromo-2-hydrazine-1,2-dihydropyridine according to step 1 in Example 1.

[0088] Step 2: Preparation of intermediate 6-bromo-3-ethyl-[1,2,4]triazolo[4,3-a]pyridine

[0089] Chemical structural formula:

[0090] Intermediate 5-bromo-2-hydrazide-1,2-dihydropyridine (2728 mg, 12.1 mmol) and propionyl chloride (25 mL) were added to a round-bottom flask and reacted at 60 °C. After the reaction was completed by TLC monitoring, the mixture was cooled to room temperature, and n-hexane (15 mL) was added. The mixture was stirred at room temperature for 15 min, filtered, and dried under infrared radiation. The crude product was dissolved in a two-phase mixture containing H₂O (20 mL) and DCM (20 mL), and 1 N NaOH solution was added to adjust the pH to neutral. The mixture was stirred at room temperature for 15 min, and the DCM layer was collected. The aqueous layer was extracted with dichloromethane. The organic layer was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. 1054.62 mg of a grayish-white solid was given, with a yield of 38.61%. 1 HNMR (300MHz, DMSO-d6) δ8.79(s,1H),7.71(d,J=9.7Hz,1H),7.43(d,J=9.6Hz,1H),3.07(q,J=7.5Hz,2H),1.34(t,J=7.5Hz,3H).

[0091] Step 3: Preparation of intermediate 3-ethyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxaldehyde

[0092] Chemical structural formula:

[0093] Under anhydrous and oxygen-free conditions, the intermediate 6-bromo-3-ethyl-[1,2,4]triazolo[4,3-a]pyridine (1000 mg, 5.7 mmol) was dissolved in anhydrous THF (14 mL), and the mixture was cooled to -8 °C. 2M i-prMgCl₂ (8 mL, 6.84 mmol) was dissolved in THF (2 mL) and added dropwise over five minutes. The reaction mixture was transferred to an ice bath and stirred for 30 minutes. DMF (1.06 mL, 13.7 mmol) was added, and the mixture was heated to 55 °C. After the reaction was complete as monitored by TLC, the reaction mixture was cooled to room temperature, and dichloromethane (10 mL) was added. The reaction mixture was then slowly poured into a 10 wt% citric acid aqueous solution (12 mL), and the two phases were stirred. The aqueous layer was extracted with dichloromethane, and the organic layer was collected, washed with saturated brine, and dried over anhydrous sodium sulfate. Purified by silica gel column chromatography (DCM:MeOH = 60:1, v / v), 446.5 mg of a grayish-white solid was obtained, with a yield of 44.70%. 1 H NMR (300MHz, DMSO-d6) δ9.97(s,1H),9.29(s,1H),7.79(d,J=9.5Hz,1H),7.60(d,J=9.6Hz,1H),3.16(q,J=7.5Hz,2H),1.41(t,J=7.5Hz,3H).

[0094] Steps 4 to 8 are the same as steps 4 to 8 in Example 1.

[0095] Step 9: Preparation of 2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-fluorophenyl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-20)

[0096] Chemical structural formula:

[0097] Under anhydrous and anaerobic conditions, Sc(OTf)3 (Bide, BD31937, 40.4 mg, 0.08 mmol) was added to an anhydrous DCM / MeOH (3 mL / 1 mL) solution of intermediates 3-ethyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxaldehyde (66.1 mg, 0.4 mmol) and 5-morpholinopyrazine-2-amine (74.4 mg, 0.4 mmol). The mixture was stirred at room temperature for 30 min. Then, intermediate 1-fluoro-4-isocyanuric acid (70.2 mg, 0.4 mmol) was added to the flask, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The reaction was quenched with saturated sodium bicarbonate solution, extracted with DCM, and purified by silica gel column chromatography (DCM:MeOH = 20:1, v / v) to give 34.5 mg of a yellowish-white solid, with a yield of 20.5%. 1H NMR (300MHz, DMSO-d6) δ9.02(s,1H),8.91(s,1H),8.74(s,1H),7.99(d,J=9.7Hz,1H),7.75(d,J=9.6Hz,1H),7.46-7.34(m,2H),7.33-7.18(m,1H) ,6.97(t,J=8.7Hz,2H),3.70(d,J=4.8Hz,4H),3.23(d,J=4.8Hz,4H),3.02(p,J=7.4,6.9Hz,2H),1.19(q,J=7.1Hz,3H).HRMS(ESI+)m / zcalculated for C 24 H 23 FN8O[M+H] + ,459.1979; found,459.1979.

[0098] 2. Synthesis of N-(4-chlorophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-21)

[0099] By replacing the intermediate 1-fluoro-4-isocyanate with 0.4 mmol of the intermediate 1-chloro-4-isocyanate, 33.2 mg of a white solid was prepared by the above method, with a yield of 18.1%. 1 H NMR (300MHz, DMSO-d6) δ8.99(s,1H),8.55(s,1H),8.06-7.94(m,2H),7.81(d,J=9.7Hz,1H),7.64(d,J=7.9Hz,1H),7.38(s,1H),7.05(t,J=7.8Hz, 1H),6.75(t,J=7.5Hz,1H),6.13(d,J=8.1Hz,1H),3.74(d,J=5.5Hz,4H),3.28(s,4H),2.98(q,J=7.5Hz,2H),1.26(t,J=7.4Hz,3H).HRMS(ESI+)m / z calculated forC 24 H 23 ClN8O[M+H] + ,475.1683;found,475.1683.

[0100] 3. Synthesis of N-(4-bromophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-22)

[0101] By replacing 4-chloroaniline with 1 mmol of 4-bromoaniline, 34.3 mg of a pale yellow solid was prepared according to the above method, with a yield of 19.2%. 1 H NMR (300MHz, DMSO-d6) δ8.99(s,1H),8.55(s,1H),8.06-7.94(m,2H),7.81(d,J=9.6Hz,1H),7.64(d,J=7.9Hz,1H),7.38(s,1H),7.05(t,J=7. 7Hz,1H),6.75(t,J=7.4Hz,1H),6.13(d,J=8.1Hz,1H),3.75(s,4H),3.29(s,4H),2.98(q,J=7.6Hz,2H),1.26(t,J=7.4Hz,3H).HRMS(ESI+)m / z calculated for C 24 H 23 BrN8O[M+H] + ,519.1178;found,519.1178.

[0102] 4. Synthesis of N-(3-bromo-4-fluorophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-23)

[0103] By replacing 4-chloroaniline with 1 mmol of 3-bromo-4-fluoroaniline, 32.3 mg of a yellow solid was prepared according to the above method, with a yield of 18.7%. 1 H NMR (300MHz, DMSO-d6) δ8.97(s,1H),8.88(s,1H),8.73(s,1H),7.95(d,J=9.7Hz,1H),7.81(d,J=9.7Hz,1H),7.49(s,1H),6.89 (p,J=8.0Hz,2H),6.19(d,J=9.5Hz,1H),3.74(s,4H),3.29(s,4H),3.03(q,J=7.7Hz,2H),1.26(t,J=7.2Hz,3H).HRMS(ESI+)m / z calculated for C 24 H 22 BrFN8O[M+H] + ,537.1084;found,537.1084.

[0104] 5. Synthesis of N-(2,4-dichlorophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-24)

[0105] By replacing 4-chloroaniline with 1 mmol of 2,4-bromoaniline, 23.9 mg of a red solid was prepared according to the above method, with a yield of 15.4%. 1 H NMR (300MHz, DMSO-d6) δ9.18(s,1H),8.99(d,J=12.9Hz,2H),8.25(d,J=9.3Hz,1H),7.83(d,J=9.3Hz,1H),7.41-7.31(m,2H),6 .91(s,1H),6.49(d,J=8.9Hz,1H),3.75(s,4H),3.28(d,J=5.8Hz,4H),2.84(q,J=7.7Hz,2H),1.35-1.22(m,3H).HRMS(ESI+)m / z calculated for C 24 H 22 Cl2N8O[M+H] + ,509.1294;found,509.1294.

[0106] 6. Synthesis of 2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-methoxyphenyl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-25)

[0107] By replacing 4-chloroaniline with 1 mmol of 1-methoxy-4-aniline, 33.6 mg of a pale red solid was prepared according to the above method, with a yield of 19.9%. 1 H NMR (300MHz, DMSO-d6) δ8.94(s,1H),8.59(s,1H),8.12(s,1H),8.03(d,J=9.7Hz,1H),7.80(d,J=9.8Hz,1H),7.34(s,1H),6.81(d,J =8.3Hz,2H),6.56(d,J=8.4Hz,2H),3.76(s,4H),3.65(s,3H),3.35(s,4H),3.02-2.93(m,2H),1.25(d,J=7.3Hz,3H).HRMS(ESI+)m / z calculated for C 25 H 26 N8O2[M+H] + ,471.2179;found,471.2179.

[0108] Example 4: Synthesis of compounds J-26-27

[0109] 1. Synthesis of 2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-fluorophenyl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-26)

[0110] Chemical structural formula:

[0111] Under anhydrous and anaerobic conditions, Sc(OTf)3 (Bidet, BD31937, 40.4 mg, 0.08 mmol) was added to anhydrous DCM / MeOH (3 mL / 1 mL) solutions of intermediates 6-bromo-3-ethyl-[1,2,4]triazolo[4,3-a]pyridine (66.1 mg, 0.4 mmol) and 5-(4-methylpiperazin-1-yl)pyrazin-2-amine (69.4 mg, 0.4 mmol). The mixture was stirred at room temperature for 30 min. Then, intermediate 1-fluoro-4-isocyanuric benzene (70.2 mg, 0.4 mmol) was added to the flask, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The reaction was quenched with saturated sodium bicarbonate solution, extracted with DCM, and purified by silica gel column chromatography with DCM / MeOH (20 / 1 + 1% TEA) to give 31.5 mg of a yellowish-white solid, with a yield of 19.2%. 1 H NMR (300MHz, DMSO-d6) δ9.05(s,1H),8.58(s,1H),8.06(s,1H),7.86(d,J=9.7Hz,1H),7.69(d,J=7.9Hz,1H),7.56(s,1H),7.37(s,1H),7.10(s,1 H),6.81(d,J=8.8Hz,1H),6.18(d,J=7.8Hz,1H),3.03(d,J=5.5Hz,4H),2.66(d,J=5.5Hz,7H),2.84(q,J=7.7Hz,2H),1.31(m,3H).HRMS(ESI+)m / z calculated for C 25 H 26 FN9[M+H] + ,472.2295;found,472.2295.

[0112] 2. Synthesis of N-(4-bromophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-27)

[0113] By replacing 4-fluoroaniline with 1 mmol of 4-bromoaniline, 25.6 mg of a pale yellow solid was prepared according to the above method, with a yield of 15.4%. 1 H NMR (300MHz, DMSO-d6) δ8.96(s,1H),8.63(s,1H),8.55(s,1H),8.14(d,J=9.9Hz,1H),7.79(d,J=9.9Hz,1H),7.52-7.46(m,2H),7.37 -7.31(m,2H),7.08(s,1H),3.77(s,4H),3.67-3.60(m,4H),3.26(s,3H)2.95(q,J=7.1Hz,2H),1.36(t,J=7.0Hz,3H).HRMS(ESI+)m / z calculated for C 25 H 26 BrN9[M+H] + ,532.1495;found,532.1495.

[0114] 3. Synthesis of N-(3-chloro-4-fluorophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-28)

[0115] By replacing the intermediate 1-fluoro-4-isocyanonitrile with 0.4 mmol of the intermediate 1-fluoro-2-chloro-4-isocyanonitrile, 31.3 mg of a pale yellow solid was prepared by the above method, with a yield of 17.6%. 1 H NMR(300MHz,DMSO-d6)δ9.09(s,1H),8.63(s,1H),8.14(d,J=9.9Hz,1H),8.1 2(s,1H),7.79(d,J=9.9Hz,1H),7.55(d,J=1.9Hz,1H),7.38(d,J=7.8Hz,1H) ,7.25(d,J=7.8Hz,1H),7.09(s,1H),3.65(s,4H),3.60(s,3H),3.35(s,4H),2.95(q,J=7.1Hz,2H),1.36(t,J=7.1Hz,3H).HRMS(ESI+)m / zcalculatedfor C 25 H 25 ClFN9[M+H] + ,506.1905;found,506.1905.

[0116] 4. Synthesis of 2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-methoxyphenyl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-29)

[0117] The intermediate 1-fluoro-4-isocyanate was replaced with 0.4 mmol of 1-methoxy-4-isocyanate in the manner described above, and 19.4 mg of a pale yellow solid was obtained with a yield of 13.1%. 1 H NMR (300MHz, DMSO-d6) δ9.05(s,1H),8.63(s,1H),8.54(s,1H),8.14(d,J=9.9,1.3Hz,1H),7.79(d,J=9.9Hz,1H),7.47-7.40(m,2H),7.08(s,1 H),6.99-6.93(m,2H),3.78(s,4H),3.77(s,3H),3.65(s,3H),3.37-3.30(m,4H),2.95(q,J=7.1Hz,2H),1.36(t,J=7.0Hz,3H).HRMS(ESI+)m / z calculated forC 26 H 29 N9O[M+H] + ,484.2495;found,484.2495.

[0118] Example 5: Synthesis of N-(4-fluorophenyl)-6-(1-methyl-1H-pyrazole-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-30)

[0119] Synthesis route:

[0120]

[0121] Steps 1 to 5 are the same as steps 1 to 5 in Example 1.

[0122] Step 6: Preparation of intermediate 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoron-2-yl)-1H-pyrrole

[0123] Chemical structural formula:

[0124] Under anhydrous and oxygen-free conditions, 4-bromo-1-methyl-1H-pyrazole (Jiuding Chemical, B140A, 2000.0 mg, 12.4 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxorane) (Bide, BD14304, 3783.1 mg, 14.9 mmol), AcOK (potassium acetate, 1828.6 mg, 18.6 mmol), Pd(dppf)Cl2 (dichloro[1,1'-bis(diphenylphosphine)ferrocene]palladium, 454.4 mg, 0.62 mmol), and dioxane (dioxane, 30 mL) were added sequentially to a two-necked flask. After the addition was complete, the temperature was naturally raised to 85 °C and the reaction was stirred. The reaction was monitored by TLC until it was complete. Heating was stopped and the mixture was cooled to room temperature. The diatomaceous earth was filtered and purified by silica gel column chromatography (DCM:MeOH = 60:1, v / v) to give 2462.9 mg of a light brown solid, with a yield of 95.5%. 1 H NMR (300MHz, DMSO-d6) δ7.89(s,1H),7.62–7.47(m,1H),3.83(s,3H),1.24(s,12H).

[0125] Step 7: Preparation of intermediate 5-(1-methyl-1H-pyrazol-4-yl)pyrazin-2-amine

[0126] Chemical structural formula:

[0127] Under nitrogen protection, 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)-1H-pyrrole (2000.0 mg, 11.5 mmol), 5-bromopyrazin-2-amine (Bide, BD21669, 2871.4 mg, 13.8 mmol), K2CO3 (4768.2 mg, 34.5 mmol), Pd(dppf)Cl2 (420.7 mg, 0.575 mmol), EtOH (ethanol, 15 mL), and water (5 mL) were added to a two-necked flask. After the addition was complete, the mixture was naturally heated to 80 °C and stirred. The reaction was monitored by TLC until complete, then heating was stopped and the mixture was cooled to room temperature. After silica gel pretreatment (1 mL of water was added to every 10 g of silica gel and stirred until homogeneous), the product was purified by silica gel column chromatography (DCM:MeOH = 80:1, v / v) to obtain 1887.1 mg of a light brown solid, with a yield of 93.7%. 1 H NMR (300MHz, DMSO-d6) δ8.24(s,1H),8.07(s,1H),7.85(d,J=7.8Hz,2H),6.35(s,2H),3.85(s,3H).

[0128] Step 8: Preparation of N-(4-fluorophenyl)-6-(1-methyl-1H-pyrazole-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-30)

[0129] Chemical structural formula:

[0130] Under anhydrous and anaerobic conditions, Sc(OTf)3 (40.4 mg, 0.08 mmol) was added to an anhydrous DCM / MeOH (3 mL / 1 mL) solution containing 3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxaldehyde (66.1 mg, 0.38 mmol) and 5-(1-methyl-1H-pyrazol-4-yl)pyrazin-2-amine (70.2 mg, 0.38 mmol). The mixture was stirred at room temperature for 30 min. Then, the intermediate 1-fluoro-4-isocyanate benzene (74.4 mg, 0.38 mmol) was added to the flask, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The reaction was quenched with saturated NaHCO3, extracted with DCM, and purified by silica gel column chromatography (DCM:MeOH = 20:1, v / v, 1% TEA) to give 24.5 mg of off-white solid, with a yield of 10.2%. 1 H NMR (300MHz, DMSO-d6) δ9.14(s,1H),8.64(s,1H),8.44(d,J=12.4Hz,2H),8.28(s,1H),7.98(d,J=14.2Hz,2H),7. 79(d,J=9.7Hz,1H),7.01(t,J=8.6Hz,2H),6.62(dd,J=8.5,4.5Hz,2H),3.87(s,3H),2.61(s,3H).HRMS(ESI+)m / z calculated forC 23 H 19 FN9[M+H] + ,440.1742; found,440.1735.

[0131] Example 6 Synthesis of 2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-fluorophenyl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-56)

[0132] Chemical structural formula:

[0133] Under anhydrous and anaerobic conditions, Sc(OTf)3 (40.4 mg, 0.08 mmol) was added to an anhydrous DCM / MeOH (3 mL / 1 mL) solution of 3-ethyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxaldehyde (66.1 mg, 0.38 mmol) and 5-(1-methyl-1H-pyrazol-4-yl)pyrazin-2-amine (70.2 mg, 0.38 mmol). The mixture was stirred at room temperature for 30 min. Then, the intermediate 1-fluoro-4-isocyanuric benzene (74.4 mg, 0.38 mmol) was added to the flask, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The reaction was quenched with saturated sodium bicarbonate solution, extracted with DCM, and purified by silica gel column chromatography (DCM:MeOH = 20:1, v / v, 1% TEA) to give 27.4 mg of off-white solid, with a yield of 10.6%. 1 H NMR(300MHz,DMSO-d6)δ9.13(s,1H),8.65(d,J=12.6Hz,2H),8.45(s,1H),8.28(s,1H),8.00(s,2H),7.80(d ,J=9.6Hz,1H),7.00(s,2H),6.64(s,2H),3.87(s,3H),3.00(s,2H),1.25(s,3H).HRMS(ESI+)m / zcalculated for C 24 H 21 FN9[M+H]+,454.1898; found,454.1901.

[0134] Example 7 Synthesis of N-(4-chlorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-2)

[0135] Chemical structural formula:

[0136] The intermediate N-(1-chloro-4-benzene)formamide was prepared by replacing 4-fluoroaniline in step 4 of Example 1 with 1 mmol of 4-chloroaniline, following the method in step 4 of Example 1. The intermediate 1-chloro-4-isocyanuric acid was then prepared by replacing N-(4-fluorobenzene)formamide in step 5 of Example 1 with 0.5 mmol of N-(1-chloro-4-benzene)formamide, following the method in step 5 of Example 1.

[0137] The intermediate 1-fluoro-4-isocyanate was replaced with 0.4 mmol of 1-chloro-4-isocyanate in the preparation method of step 9 in Example 1, and 24.5 mg of a pale yellow solid was obtained with a yield of 20%. 1H NMR (300MHz, DMSO-d6) δ8.96(s,1H),8.64(d,J=7.5Hz,2H),7.95(d,J=9.7Hz,1H),7.79(d,J=9.8Hz,1H),7.30(s,1H), 7.21(d,J=8.3Hz,2H),6.60(d,J=8.3Hz,2H),3.75(d,J=4.6Hz,4H),3.26(d,J=4.6Hz,4H),2.63(s,3H).HRMS(ESI+)m / z calculated for C 23 H 21 ClN8O[M+H] + ,461.1527;found,461.1527.

[0138] Example 8: Synthesis of N-(3-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-3)

[0139] Chemical structural formula:

[0140] The intermediate N-(1-fluoro-3-benzene)formamide was prepared by replacing 4-fluoroaniline in step 4 of Example 1 with 1 mmol of 3-fluoroaniline, following the method in step 4 of Example 1. The intermediate 1-fluoro-3-isocyanuric acid was prepared by replacing N-(4-fluorobenzene)formamide in step 5 of Example 1 with 0.5 mmol of N-(1-fluoro-3-benzene)formamide, following the method in step 5 of Example 1.

[0141] The intermediate 1-fluoro-3-isocyanate benzene was replaced with 0.4 mmol of the intermediate 1-fluoro-4-isocyanate benzene, and the preparation method in step 9 of Example 1 was used to prepare 44.5 mg of a light red solid, with a yield of 31.2%. 1 H NMR (300MHz, DMSO-d6) δ8.99(s,1H),8.61(d,J=7.5Hz,2H),7.95(d,J=9.7Hz,1H),7.79(d,J=9.7Hz,1H),7.48(d,J=7.9Hz,1H),7.37(s,1H),7.01( d,J=8.0Hz,1H),6.81(d,J=7.7Hz,1H),6.15(d,J=8.1Hz,1H),3.75(d,J=4.6Hz,4H),3.29(d,J=4.6Hz,4H),2.62(s,3H).HRMS(ESI+)m / zcalculated for C 23 H 21FN8O[M+H] + ,445.1822;found,445.1822.

[0142] Example 9: Synthesis of compounds J-4 to J-8

[0143] 1. Synthesis of N-(2-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-4)

[0144] Chemical structural formula:

[0145] The intermediate N-(1-fluoro-2-benzene)formamide was prepared by replacing 4-fluoroaniline in step 4 of Example 1 with 1 mmol of 2-fluoroaniline, following the method in step 4 of Example 1. The intermediate 1-fluoro-2-isocyanuric acid was prepared by replacing N-(4-fluorobenzene)formamide in step 5 of Example 1 with 0.5 mmol of N-(1-fluoro-2-benzene)formamide, following the method in step 5 of Example 1.

[0146] The intermediate 1-fluoro-2-isocyanate benzene was replaced with 0.4 mmol of the intermediate 1-fluoro-4-isocyanate benzene, and the preparation method in step 9 of Example 1 was used to prepare a red solid of 34.5 mg, with a yield of 24.2%. 1 H NMR(300MHz,DMSO-d6)δ8.98(s,1H),8.59(s,1H),8.08(s,1H),7.95(d,J= 9.6Hz,1H),7.78(d,J=9.6Hz,1H),7.62(d,J=8.0Hz,1H),7.37(s,1H),7.04 (d,J=7.7Hz,1H),6.73(d,J=7.6Hz,1H),6.13(d,J=8.1Hz,1H),3.75(d,J=4.6Hz,4H),3.27(d,J=4.6Hz,4H),2.61(s,3H).HRMS(ESI+)m / zcalculated for C 23 H 21 FN8O[M+H] + ,445.1822;found,445.1822.

[0147] 2. Synthesis of N-(3-chloro-4-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-5)

[0148] By replacing 4-fluoroaniline with 1 mmol of 1-fluoro-2-chloro-4-aniline, 22.5 mg of a pale red solid was prepared according to the method described above, with a yield of 12.4%. 1 H NMR (300MHz, DMSO-d6) δ8.96(s,1H),8.65(s,1H),8.56(s,1H),7.95(d,J=9.7Hz,1H),7.80(d,J=9.6Hz,1H),7.33(s,1H),7.21 (d,J=9.0Hz,1H),6.78(d,J=6.1Hz,1H),6.49(s,1H),3.75(d,J=4.7Hz,4H),3.29(d,J=4.8Hz,4H),2.66(s,3H).HRMS(ESI+)m / z calculated for C 23 H 20 ClFN8O[M+H] + ,479.1433;found,479.1433.

[0149] 3. Synthesis of N-(2,4-dichlorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-6)

[0150] By replacing 4-fluoroaniline with 1 mmol of 1,3-dichloro-4-aniline, 33.4 mg of a pale red solid was prepared according to the above method, with a yield of 21.4%. 1 H NMR (300MHz, DMSO-d6) δ9.61(s,1H),9.08(s,1H),8.97(s,1H),8.09(d,J=9.7Hz,1H),7.91(d,J=9.6Hz,1H),7.54 -7.43(m,2H),7.10(s,1H),6.64(d,J=8.1Hz,1H),3.84(s,4H),3.40(d,J=4.8Hz,4H),2.82(s,3H).HRMS(ESI+)m / z calculatedfor C 23 H 20 Cl2N8O[M+H] + ,495.1137; found,495.113.

[0151] 4. Synthesis of N-(2,4-difluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinimidazo[1,2-a]pyrazin-3-amine (J-7)

[0152] By replacing 4-fluoroaniline with 1 mmol of 1,3-difluoro-4-aniline, 19.4 mg of a pale red solid was prepared according to the above method, with a yield of 12.3%. 1 H NMR (300MHz, DMSO-d6) δ8.97(s,1H),8.61(s,1H),8.03(s,1H),7.94(d,J=9.7Hz,1H),7.79(d,J=9.8Hz,1H),7.67–7.57(m,1H), 7.39(s,1H),7.00–6.88(s,1H),6.16(d,J=5.1Hz,1H),3.75(d,J=5.5Hz,4H),3.39(d,J=4.8Hz,4H),2.65(s,3H).HRMS(ESI+)m / z calculated for C 23 H 20 F2N8O[M+H] + ,463.1728;found,463.1728.

[0153] 5. Synthesis of 2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholino-N-(4-(trifluoromethyl)phenyl)imidazo[1,2-a]pyrazin-3-amine (J-8)

[0154] By replacing 4-fluoroaniline with 1 mmol of 1-trifluoromethyl-4-aniline, 49.4 mg of a yellow solid was prepared according to the above method, with a yield of 34.2%. 1 H NMR (300MHz, DMSO-d6) δ9.17(s,1H),9.06(s,1H),9.00(s,1H),8.26(d,J=9.4Hz,1H),7.82(d,J=9.3Hz,1H),7.53(d,J=8. 4Hz,2H),7.32(s,1H),6.75(d,J=8.2Hz,2H),3.76(t,J=4.6Hz,4H),3.28(s,4H),2.48(s,3H).HRMS(ESI+)m / zcalculated for C 24 H 20 F3N8O[M+H] + ,495.1790; found,495.1790.

[0155] Example 10: Synthesis of N-(4-methoxyphenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-morpholinylimidazo[1,2-a]pyrazin-3-amine (J-9)

[0156] Chemical structural formula:

[0157] The intermediate N-(1-methoxy-4-benzene)formamide was prepared by replacing 4-fluoroaniline in step 4 of Example 1 with 1 mmol of 1-methoxy-4-aniline, following the method in step 4 of Example 1. The intermediate 1-methoxy-4-isocyanuric acid was then prepared by replacing N-(4-fluorobenzene)formamide in step 5 of Example 1 with 0.5 mmol of N-(1-methoxy-4-benzene)formamide, following the method in step 5 of Example 1.

[0158] The intermediate 1-fluoro-4-isocyanate was replaced with 0.4 mmol of 1-methoxy-4-isocyanate in the manner described in step 9 of Example 1, and 29.4 mg of yellow solid was obtained, with a yield of 18.7%. 1 H NMR (300MHz, DMSO-d6) δ8.93(s,1H),8.61(s,1H),8.19(s,1H),8.00(d,J=9.7Hz,1H),7.78(d,J=9.7Hz,1H),7.40(s,1H),6. 95(s,1H),6.79(d,J=8.4Hz,2H),6.55(d,J=8.4Hz,2H),3.64(s,3H),3.52(s,3H),2.82(s,4H),2.60(s,3H).HRMS(ESI+)m / z calculated for C 24 H 24 N8O2[M+H] + ,457.2022;found,457.2022.

[0159] Example 11 Synthesis of compounds J-15-16

[0160] 1. Synthesis of N-(3-chloro-4-fluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-15)

[0161] Chemical structural formula:

[0162] Intermediate N-(1-fluoro-2-chloro-4-benzene)formamide was prepared by replacing 4-fluoroaniline with 1 mmol of 1-fluoro-2-chloro-4-aniline according to step 4 of Example 1; intermediate 1-fluoro-2-chloro-4-isocyanuric acid was prepared by replacing N-(4-fluorobenzene)formamide with 0.5 mmol of N-(1-fluoro-2-chloro-4-benzene)formamide according to step 5 of Example 1.

[0163] The intermediate 1-fluoro-2-chloro-4-isocyanonitrile was replaced with 0.4 mmol of the intermediate 1-fluoro-4-isocyanonitrile, and the preparation method of Example 2 was used to prepare the product, yielding 48.3 mg of off-white solid with a yield of 29.2%. 1 H NMR (300MHz, DMSO-d6) δ8.83(s,1H),8.63(s,1H),8.15(d,J=1.3Hz,1H),8.13(d,J=4.2Hz,1H),7. 75(s,2H),7.55(d,J=1.9Hz,1H),7.25(d,J=7.8Hz,1H),7.09(s,1H),3.65(d,J=5.5Hz,4H),3.60(d J=5.6Hz,4H),2.43(s,3H),2.23(s,3H).HRMS(ESI+)m / z calculated for C 24 H 23 ClFN9[M+H] + ,492.1749;found,492.1749.

[0164] 2. Synthesis of N-(2,4-dichlorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-16)

[0165] The red solid was prepared using the method described above, yielding 32.7 mg in a yield of 22.3%. 1 H NMR (300MHz, DMSO-d6) δ8.83(s,1H),8.71(s,1H),8.63(s,1H),8.14(d,J=9.9Hz,1H),7.75(d,J=9.7Hz,1H),7.54–7.47(m,2H),7.37(d J=8.1Hz,1H),7.09(s,1H),3.65(d,J=5.5Hz,2H),3.60(d,J=5.6Hz,2H),3.43(s,4H),2.64(s,3H),2.23(s,3H).HRMS(ESI+)m / z calculated forC 24 H 23 Cl2N9[M+H] + ,508.1453;found,508.1453.

[0166] Example 12: Synthesis of compounds J-17 to J-18

[0167] 1. Synthesis of N-(2,4-difluorophenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-17)

[0168] Chemical structural formula:

[0169] Intermediate N-(1,3-difluoro-4-benzene)formamide was prepared by replacing 4-fluoroaniline with 1 mmol of 1,3-difluoro-4-aniline according to step 4 of Example 1; intermediate 1,3-difluoro-4-isocyanate was prepared by replacing N-(4-fluorobenzene)formamide with 0.5 mmol of N-(1,3-difluoro-4-benzene)formamide according to step 5 of Example 1.

[0170] The intermediate 1-fluoro-4-isocyanate was replaced with 0.4 mmol of intermediate 1,3-difluoro-4-isocyanate in the manner described in Example 2, and 18.3 mg of a pale red solid was obtained with a yield of 10.2%. 1 H NMR (300MHz, DMSO-d6) δ8.88(s,1H),8.63(s,1H),8.25(s,1H),8.14(dd,J=9.9Hz,1H),7.75(d,J=9.7Hz,1H),7.51(d,J =7.3Hz,1H),7.09(s,1H),7.07-7.02(m,2H),3.27(d,J=5.5Hz,4H),2.73(d,J=5.5Hz,7H),2.22(s,3H).HRMS(ESI+)m / z calculated for C 24 H 23 F2N9[M+H] + ,476.2044;found,476.2044.

[0171] 2. Synthesis of 2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)-N-(4-(trifluoromethyl)phenyl)imidazo[1,2-a]pyrazin-3-amine (J-18)

[0172] By replacing 4-fluoroaniline with 1 mmol of 1-trifluoromethyl-4-aniline, 38.3 mg of a red solid was prepared according to the above method, with a yield of 19.3%. 1H NMR (300MHz, DMSO-d6) δ8.87(s,1H),8.63(s,1H),8.52(s,1H),8.14(d,J=9.9Hz,1H),7.75(d,J=9.7Hz,1H),7.68-7.58(m,4H) ,7.09(s,1H),3.65(d,J=5.5Hz,2H),3.60(d,J=5.6Hz,2H),2.73(s,4H),2.38(s,3H),2.23(s,3H).HRMS(ESI+)m / zcalculated for C 25 H 24 F3N9[M+H] + ,508.2107;found,508.2107.

[0173] 3. Synthesis of N-(4-methoxyphenyl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(4-methylpiperazin-1-yl)imidazo[1,2-a]pyrazin-3-amine (J-19)

[0174] By replacing the intermediate 1-fluoro-4-isocyanate with 0.4 mmol of the intermediate 1-methoxy-4-isocyanate, 16.2 mg of a white solid was prepared by the above method, with a yield of 9.1%. 1 H NMR (300MHz, DMSO-d6) δ8.84(s,1H),8.54(s,1H),8.14(d,J=9.9Hz,1H),7.75(d,J=9.7Hz,1H),7.19(d,J=1.4Hz,1H),7.05(d,J=7.8H z,1H),7.09(s,1H),6.71(d,J=7.7Hz,1H),6.58(m,2H),3.78(s,3H),3.65(d,J=2.7Hz,2H),3.60(d,J=2.8Hz,2H),2.73(s,4H),2.64(d J=2.7Hz,3H),2.21(s,3H).HRMS(ESI+)m / z calculated for C 25 H 27 N9O[M+H] + ,470.2339;found,470.2339.

[0175] Example 13: Synthesis of compounds J-31 to 34

[0176] 1. Synthesis of N-(4-chlorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-31)

[0177] Chemical structural formula:

[0178] The intermediate 1-chloro-4-isocyanonitrile was synthesized according to the method of Example 7, with 0.4 mmol of intermediate 1-chloro-4-isocyanonitrile replacing intermediate 1-fluoro-4-isocyanonitrile. The preparation method of Example 5 was used to obtain 61.4 mg of a pale yellow solid, with a yield of 23.6%. 1 H NMR (300MHz, DMSO-d6) δ9.15(d,J=1.5Hz,1H),8.63(d,J=15.4Hz,2H),8.41(d,J=1.6Hz,1H),8.29(s,1H),8.04-7.91 (m,2H),7.80(d,J=9.6Hz,1H),7.21(d,J=8.3Hz,2H),6.63(d,J=8.4Hz,2H),3.88(s,3H),2.63(s,3H).HRMS(ESI+)m / z calculated forC 23 H 19 ClN9[M+H] + ,456.1446; found,456.1440.

[0179] 2. Synthesis of N-(4-bromophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-32)

[0180] By replacing the intermediate 1-fluoro-4-isocyanate with 0.4 mmol of the intermediate 1-bromo-4-isocyanate, 71.5 mg of a pale yellow solid was prepared by the above method, with a yield of 25.1%. 1 H NMR (300MHz, DMSO-d6) δ9.10(d,J=22.1Hz,2H),8.79(s,1H),8.44(s,1H),8.39-8.19(m,1H),7.98(d,J=16.4Hz,2H),7. 79(d,J=9.7Hz,1H),7.30(d,J=8.2Hz,2H),6.60(d,J=8.3Hz,2H),3.87(s,3H),2.65(s,3H).HRMS(ESI+)m / zcalculated for C 23 H19 BrN9[M+H] + ,500.0941; found,500.0947.

[0181] 3. Synthesis of N-(3-fluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-33)

[0182] The intermediate 1-fluoro-2-chloro-4-isocyanonitrile was synthesized according to the method of Example 8, with 0.4 mmol of intermediate 1-fluoro-3-isocyanonitrile used instead of intermediate 1-fluoro-4-isocyanonitrile. The preparation method of Example 5 was used to obtain 43.3 mg of a pale yellow solid, with a yield of 17.3%. 1 H NMR (300MHz, DMSO-d6) δ9.24(s,1H),9.14(s,1H),8.80(s,1H),8.47(s,1H),8.30(s,1H),7.99(d,J=11.5Hz,2H),7.79(d,J=9.9H z,1H),7.15(d,J=8.1Hz,1H),6.52(d,J=11.0Hz,2H),6.44(d,J=8.1Hz,1H),3.87(s,3H),2.65(s,3H).HRMS(ESI+)m / zcalculated for C 23 H 19 FN9[M+H] + ,440.1742; found,440.1737.

[0183] 4. Synthesis of N-(2-fluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-34)

[0184] By replacing 4-fluoroaniline with 1 mmol of 2-fluoroaniline, 73.3 mg of a pale yellow solid was prepared according to the above method, with a yield of 29.4%. 1H NMR (300MHz, DMSO-d6) δ9.17(s,1H),8.60(s,1H),8.50(s,1H),8.28(d,J=26.1Hz,2H),8.04(s,1H),7.93(d,J=9.7Hz,1H),7.79(d,J=9.7H z,1H),7.48(d,J=7.9Hz,1H),6.98(t,J=7.6Hz,1H),6.79(t,J=7.7Hz,1H),6.20(d,J=8.1Hz,1H),3.87(s,3H),2.60(s,3H).HRMS(ESI+)m / z calculated for C 23 H 19 FN9[M+H] + ,440.1742; found,440.1745.

[0185] 5. Synthesis of N-(3-chloro-4-fluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-35)

[0186] The intermediate 1-fluoro-2-chloro-4-isocyanonitrile was synthesized according to the method of Example 17, with 0.4 mmol of the intermediate 1-fluoro-2-chloro-4-isocyanonitrile used instead of the intermediate 1-fluoro-4-isocyanonitrile. The preparation method of Example 5 was used to obtain 42.6 mg of a pale yellow solid, with a yield of 15.8%. 1 H NMR (300MHz, DMSO-d6) δ9.14(d,J=1.5Hz,1H),8.65(d,J=17.8Hz,2H),8.44(d,J=1.6Hz,1H),8.29(s,1H),8.02(s,1H),7.94(d,J=9.7H z,1H),7.81(d,J=9.7Hz,1H),7.20(t,J=9.0Hz,1H),6.89-6.77(m,1H),6.53(d,J=8.7Hz,1H),3.87(s,3H),2.65(s,3H).HRMS(ESI+)m / z calculated for C 23 H 18 ClFN9[M+H] + ,474.1352;found,474.1356.

[0187] 6. Synthesis of 6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-phenylimidazo[1,2-a]pyrazin-3-amine (J-36)

[0188] By replacing 4-fluoroaniline with 1 mmol of aniline, 56.7 mg of a pale yellow solid was prepared according to the above method, with a yield of 23.6%. 1 H NMR (300MHz, DMSO-d6) δ9.13(s,1H),8.78(d,J=39.9Hz,1H),8.46(s,1H),8.27(s,1H),7.98(s,2H),7.76(d,J= 9.7Hz,1H),7.15(t,J=7.0Hz,2H),6.75(s,1H),6.64(d,J=7.9Hz,2H),3.86(s,3H),2.59(s,3H).HRMS(ESI+)m / z calculated for C 23 H 20 N9[M+H] + ,422.1836; found,422.1839.

[0189] Example 14: Synthesis of compounds J-37 to J-41

[0190] 1. Synthesis of 6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-N-(4-(trifluoromethyl)phenyl)imidazo[1,2-a]pyrazin-3-amine (J-37)

[0191] Chemical structural formula:

[0192] The intermediate 1-fluoro-4-isocyanate was replaced with 0.4 mmol of 1-trifluoromethyl-4-isocyanate in the manner described in Example 5, and 39.4 mg of a pale yellow solid was obtained, with a yield of 14.1%. 1 H NMR (300MHz, DMSO-d6) δ9.61(s,1H),9.16(s,1H),8.84(s,1H),8.47(s,1H),8.29(s,1H),7.98(d,J=13.7 Hz,2H),7.78(s,1H),7.47(s,2H),6.80(s,2H),3.86(s,3H),2.65(s,3H).HRMS(ESI+)m / zcalculatedfor C 24 H 19 F3N9[M+H] +,490.1710; found,490.1717.

[0193] 2. Synthesis of N-(2-bromo-4-fluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-38)

[0194] By replacing 4-fluoroaniline with 1 mmol of 1-fluoro-3-bromo-4-aniline, 59.0 mg of a pale yellow solid was prepared according to the above method, with a yield of 19.9%. 1 H NMR(300MHz,DMSO-d6)δ9.14(d,J=1.4Hz,1H),8.64-8.58(m,1H),8.51(d,J=1.5 Hz,1H),8.31(s,1H),8.08(s,1H),8.03(s,1H),7.92(dd,J=9.6,1.5Hz,1H),7.77 (d,J=9.7Hz,1H),7.60(dd,J=8.2,2.9Hz,1H),6.91(td,J=8.6,3.0Hz,1H),6.24(dd,J=9.0,5.1Hz,1H),3.87(s,3H),2.63(s,3H).HRMS(ESI+)m / zcalculatedfor C 23 H 18 BrFN9[M+H] + ,518.0847;found,518.0851.

[0195] 3. Synthesis of N-(2-bromo-4-fluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-39)

[0196] By replacing 4-fluoroaniline with 1 mmol of 2,6-fluoroaniline, a pale yellow solid of 67.2 mg was prepared according to the above method, with a yield of 25.8%. 1H NMR (300MHz, DMSO-d6) δ9.11(d,J=1.4Hz,1H),8.75(s,1H),8.64(s,1H),8.47(s,1H),8.31(s,1H),8.04(s,1H),7.90(d,J=9.6 Hz,1H),7.77(d,J=9.6Hz,1H),7.00(t,J=8.5Hz,2H),6.87-6.78(m,1H),3.89(s,3H),2.64(s,3H).HRMS(ESI+)m / zcalculated for C 23 H 18 F2N9[M+H] + ,458.1648;found,458.1652.

[0197] 4. Synthesis of N-(4-bromo-3-fluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-40)

[0198] Intermediate N-(1-bromo-2-fluoro-4-benzene)formamide was prepared by replacing 4-fluoroaniline with 1 mmol of 1-bromo-2-fluoro-4-aniline according to step 4 of Example 2; intermediate 1-bromo-2-fluoro-4-isocyanuric acid was prepared by replacing N-(4-fluorobenzene)formamide in step 5 of Example 1 with 0.5 mmol of intermediate N-(1-bromo-2-fluoro-4-benzene)formamide according to step 5 of Example 1.

[0199] The intermediate 1-bromo-2-fluoro-4-isocyanate was replaced with 0.4 mmol of the intermediate 1-bromo-2-fluoro-4-isocyanate, and the preparation method of Example 5 was used to prepare the product, yielding 79.1 mg of a pale yellow solid with a yield of 26.8%. 1 H NMR (300MHz, DMSO-d6) δ9.44(s,1H),9.14(s,1H),8.84(s,1H),8.45(s,1H),8.29(s,1H),7.98(d,J=21.5Hz,2H),7.79(d,J =9.6Hz,1H),7.40(t,J=8.4Hz,1H),6.71(d,J=13.2Hz,1H),6.43(d,J=8.6Hz,1H),3.87(s,3H),2.68(s,3H).HRMS(ESI+)m / z calculated for C 23 H 18 BrFN9[M+H] + ,518.0847;found,518.0846.

[0200] 5. Synthesis of N-(4-bromo-2-fluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-41)

[0201] By replacing 4-fluoroaniline with 1 mmol of 1-bromo-3-fluoro-4-aniline, 47.8 mg of a pale yellow solid was prepared according to the above method, with a yield of 16.2%. 1 H NMR(300MHz,DMSO-d6)δ9.16(s,1H),8.74(s,1H),8.66(s,1H),8.51(s,1H),8.30(s,1H),8.03(s,1H),7.90(d,J=9.6Hz,1H),7 .80(d,J=9.6Hz,1H),7.61-7.52(m,1H),7.02(d,J=8.6Hz,1H),6.21(t,J=9.0Hz,1H),3.87(s,3H),2.66(s,3H).HRMS(ESI+)m / z calculated for C 23 H 18 BrFN9[M+H] + ,518.0847; found,518.0850. Example 15 Synthesized compounds J-42~50

[0202] 1. Synthesis of N-(2-chlorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-42)

[0203] Chemical structural formula:

[0204] Intermediate N-(2-chlorobenzene)formamide was prepared by replacing 4-fluoroaniline with 1 mmol of 2-chloroaniline according to step 4 of Example 1; intermediate 1-chloro-2-isocyanuric acid was prepared by replacing N-(4-fluorobenzene)formamide with 0.5 mmol of N-(2-chlorobenzene)formamide according to step 5 of Example 1.

[0205] The intermediate 1-fluoro-4-isocyanate was replaced with 0.4 mmol of 1-chloro-2-isocyanate in place of the intermediate 1-fluoro-4-isocyanate, and the preparation method of Example 5 was used to obtain 57.4 mg of a pale yellow solid, with a yield of 21.1%. 1H NMR (300MHz, DMSO-d6) δ9.17(s,1H),8.61(s,1H),8.50(s,1H),8.29(d,J=21.1Hz,2H),8.04(s,1H),7.93(d,J=9.7Hz,1H),7.79(d,J=9.7H z,1H),7.48(d,J=7.9Hz,1H),6.98(t,J=7.7Hz,1H),6.79(t,J=7.6Hz,1H),6.20(d,J=8.1Hz,1H),3.87(s,3H),2.60(s,3H).HRMS(ESI+)m / z calculated forC 23 H 19 ClN9[M+H] + ,456.1447; found,456.1445.

[0206] 2. Synthesis of N-(2,4-chlorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-43)

[0207] The intermediate 1-fluoro-4-isocyanonitrile was replaced with 0.4 mmol of the intermediate 1,3-chloro-4-isocyanonitrile, and the preparation method of Example 5 was used to prepare the product, yielding 42.1 mg of a pale yellow solid with a yield of 15.1%. 1 H NMR(300MHz,DMSO-d6)δ9.17(d,J=1.5Hz,1H),8.67(s,1H),8.49(d,J=1.5H z,1H),8.39(s,1H),8.32(s,1H),8.04(s,1H),7.89(dd,J=9.6,1.5Hz,1H), 7.80(d,J=9.7Hz,1H),7.62(d,J=2.4Hz,1H),7.03(dd,J=8.7,2.4Hz,1H),6.21(d,J=8.8Hz,1H),3.87(s,3H),2.64(s,3H).HRMS(ESI+)m / zcalculated for C 23 H 18 C l2 N9[M+H] + ,490.1057;found,490.1058.

[0208] 3. Synthesis of N-(4-methoxyphenyl)-6-(1-methyl-1H-pyrazole-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-44)

[0209] By replacing 4-fluoroaniline with 1 mmol of 1-methoxy-4-aniline, 37.2 mg of a pale yellow solid was prepared according to the above method, with a yield of 14.4%. 1 H NMR (300MHz, DMSO-d6) δ9.15(d,J=18.9Hz,2H),8.47(d,J=24.4Hz,2H),8.28(d,J=10.1Hz,2H),7.97(s,1H),7.78(d,J=9. 3Hz,1H),6.76(d,J=8.1Hz,2H),6.57(d,J=8.3Hz,2H),3.86(s,3H),3.61(s,3H),2.45(s,3H).HRMS(ESI+)m / zcalculated for C 24 H 21 N9O[M+H] + ,452.1869;found,452.1869.

[0210] 4. Synthesis of N-(2-chloro-4-bromophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-45)

[0211] By replacing 4-fluoroaniline with 1 mmol of 1-bromo-3-chloro-4-aniline, 71.0 mg of a pale yellow solid was prepared according to the above method, with a yield of 23.3%. 1 H NMR(300MHz,DMSO-d6)δ9.16(s,1H),8.70(s,1H),8.49(s,2H),8.32(s,1H),8.04(s,1H),7.94-7.7 6(m,2H),7.70(s,1H),7.13(s,1H),6.15(d,J=8.8Hz,1H),3.87(s,3H),2.64(s,3H).HRMS(ESI+)m / z calculated for C 23 H 18 BrClN9[M+H] + ,534.0552;found,534.0557.

[0212] 5. Synthesis of N-(2-bromophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-46)

[0213] Replacing 4-fluoroaniline with 1 mmol of 2-bromoaniline, the product was prepared according to the method described above, yielding 89.3 mg of a pale yellow solid, with a yield of 31.3%. 1 H NMR (300MHz, DMSO-d6) δ9.17(d,J=1.5Hz,1H),8.57(s,1H),8.54-8.49(m,1H),8.33(s,1H),8.06(d,J=10.7Hz,2H),7.94(dd,J=9.7,1.6Hz,1H),7.7 9(d,J=9.6Hz,1H),7.67-7.61(m,1H),7.02(t,J=7.7Hz,1H),6.74(d,J=7. 6Hz,1H),6.19(d,J=8.0Hz,1H),3.87(s,3H),2.60(s,3H).HRMS(ESI+)m / z calculated for C 23 H 19 BrN9[M+H] + ,500.0947; found,500.0944.

[0214] 6. Synthesis of N-(3-bromophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-47)

[0215] Intermediate N-(3-bromobenzene)formamide was prepared by replacing 4-fluoroaniline with 1 mmol of 3-bromoaniline according to step 4 of Example 1; intermediate 1-bromo-3-isocyanate was prepared by replacing N-(4-fluorobenzene)formamide with 0.5 mmol of N-(3-bromobenzene)formamide according to step 5 of Example 1. Intermediate 1-bromo-3-isocyanate was prepared by replacing intermediate 1-fluoro-4-isocyanate with 0.4 mmol of intermediate 1-bromo-3-isocyanate using the preparation method of Example 5, yielding 37.5 mg of a pale yellow solid, with a yield of 13.2%. 1H NMR(300MHz,DMSO-d6)δ9.09(s,1H),8.65(s,1H),8.43(s,1H),8.23(s,1H),7.98(s,2H),7.76(d,J=9.7Hz,1 H),7.05(d,J=8.6Hz,1H),6.94-6.83(m,2H),6.52(d,J=8.0Hz,1H),3.84(s,3H),2.60(s,3H).HRMS(ESI+)m / z calculated for C 23 H 19 BrN9[M+H] + ,500.0947; found,500.0943.

[0216] 7. Synthesis of N-(3-chlorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-48)

[0217] By replacing 4-fluoroaniline with 1 mmol of 3-chloroaniline, 41.4 mg of a pale yellow solid was prepared according to the above method, with a yield of 15.9%. 1 H NMR (300MHz, DMSO-d6) δ9.16(d,J=12.1Hz,2H),8.79(s,1H),8.48(s,1H),8.29(s,1H),7.98(d,J=17.1Hz,2H),7.79(d,J=9.6 Hz,1H),7.13(t,J=8.2Hz,1H),6.78(d,J=7.6Hz,2H),6.57-6.47(m,1H),3.87(s,3H),2.65(s,3H).HRMS(ESI+)m / zcalculated for C 23 H 19 ClN9[M+H] + ,456.1447;found,456.1443.

[0218] 8. Synthesis of N-(3,4-dichlorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-49)

[0219] Intermediate N-(1,2-chloro-4-benzene)formamide was prepared by replacing 4-fluoroaniline with 1 mmol of 1,2-chloro-4-aniline according to step 4 of Example 2. Intermediate 1,2-chloro-4-isocyanuric acid was prepared by replacing N-(4-fluorobenzene)formamide in step 5 of Example 5 with 0.5 mmol of intermediate N-(1,2-chloro-4-benzene)formamide according to step 5 of Example 5. Intermediate 1,2-chloro-4-isocyanuric acid was prepared by replacing intermediate 1-fluoro-4-isocyanuric acid with 0.4 mmol of intermediate 1,2-chloro-4-isocyanuric acid using the preparation method of Example 5, yielding 74.8 mg of a pale yellow solid with a yield of 26.8%. 1 H NMR(300MHz,DMSO-d6)δ9.15(s,1H),8.83(s,1H),8.70(s,1H),8.45(s,1H),8.30(s,1H),8.03(s,1H),7.97-7.77(m, 2H),7.36(d,J=8.7Hz,1H),6.91(s,1H),6.53(d,J=8.5Hz,1H),3.87(s,3H),2.66(s,3H).HRMS(ESI+)m / zcalculated for C 23 H 18 C l2 N9[M+H] + ,490.1057;found,490.1052.

[0220] 9. Synthesis of N-(2-fluoro-3-chlorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-2-(3-methyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)imidazo[1,2-a]pyrazin-3-amine (J-50)

[0221] By replacing 4-fluoroaniline with 1 mmol of 1-chloro-2-fluoro-3-aniline, 45.2 mg of a pale yellow solid was prepared according to the method described above, with a yield of 16.7%. 1 H NMR(300MHz,DMSO-d6)δ9.16(s,1H),8.71(s,2H),8.55(s,1H),8.30(s,1H),8.04(s,1H),7 .86(d,J=27.7Hz,2H),6.89(s,2H),6.18(s,1H),3.87(s,3H),2.65(s,3H).HRMS(ESI+)m / z calculated for C 23 H 18 ClFN9[M+H] + ,474.1352;found,474.1351.

[0222] Example 16: Synthesis of J-51 to J-55

[0223] 1. Synthesis of N-(4-chlorophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-3-amine (J-52)

[0224] Chemical structural formula:

[0225] The intermediate 1-fluoro-4-isocyanate was replaced with 0.4 mmol of 1-chloro-4-isocyanate in place of the intermediate 1-fluoro-4-isocyanate, and the preparation method of Example 6 was used to prepare the product, yielding 71.4 mg of a pale yellow solid with a yield of 26.7%. 1 H NMR (300MHz, DMSO-d6) δ9.20-9.07(m,1H),8.86(s,1H),8.69(s,1H),8.45(s,1H),8.29(s,1H),7.99(d,J=12.3Hz,2H),7.80(d,J =9.6Hz,1H),7.19(d,J=8.4Hz,2H),6.64(d,J=8.3Hz,2H),3.87(s,3H),3.04-2.98(m,2H),1.26(d,J=7.5Hz,3H).HRMS(ESI+)m / z calculated for C 24 H 21 ClN9[M+H] + ,470.1603; found,470.1606.

[0226] 2. Synthesis of N-(4-bromophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-3-amine (J-51)

[0227] The intermediate 1-bromo-4-isocyanate was replaced with 0.4 mmol of the intermediate 1-bromo-4-isocyanate in place of the intermediate 1-fluoro-4-isocyanate, and the mixture was prepared according to the method in Example 6, yielding 50.0 mg of a pale yellow solid with a yield of 17.1%. 1H NMR(300MHz,DMSO-d6)δ9.18-9.10(m,1H),8.71(s,1H),8.64(s,1H),8.44(d,J=1.6Hz,1H),8.29(s,1H),8.01(s,1H), 7.97(d,J=9.7Hz,1H),7.81(d,J=9.6Hz,1H),3.87(s,3H),3.00(q,J=7.5Hz,2H),1.26(t,J=7.5Hz,3H).HRMS(ESI+)m / z calculated for C 24 H 21 BrN9[M+H] + ,514.1098;found,514.1102.

[0228] 3. Synthesis of N-(2-fluorophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-3-amine (J-53)

[0229] By replacing the intermediate 1-fluoro-4-isocyanate with 0.4 mmol of the intermediate 1-fluoro-2-isocyanate, 48.9 mg of a pale yellow solid was prepared by the above method, with a yield of 18.9%. 1 H NMR(300MHz,DMSO-d6)δ9.17(s,1H),8.58(s,1H),8.50(s,1H),8.32(s,1H),8 .24(s,1H),8.04(s,1H),7.97(d,J=9.7Hz,1H),7.80(d,J=9.6Hz,1H),7.49(d, J=7.9Hz,1H),6.99(t,J=7.8Hz,1H),6.80(t,J=7.7Hz,1H),6.20(d,J=8.1Hz, 1H),3.87(s,3H),2.96(q,J=7.5Hz,2H),1.25(t,J=7.4Hz,3H).HRMS(ESI+)m / z calculated for C 24 H 21 FN9[M+H] + ,454.1899; found,454.1903.

[0230] 4. Synthesis of N-(2-bromophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-3-amine (J-54)

[0231] By replacing the intermediate 1-fluoro-4-isocyanate with 0.4 mmol of the intermediate 1-bromo-2-isocyanate, 105.4 mg of off-white solid was prepared according to the above method, with a yield of 35.9%. 1 H NMR(300MHz,DMSO-d6)δ9.17(d,J=1.5Hz,1H),8.57-8.48(m,2H),8.32(s,1H),8 .05(d,J=5.0Hz,2H),7.98(d,J=9.6Hz,1H),7.81(d,J=9.7Hz,1H),7.64(d,J=7.9 Hz,1H),7.03(t,J=7.7Hz,1H),6.74(t,J=7.6Hz,1H),6.19(d,J=8.1Hz,1H),3.87 (s,3H),2.96(q,J=7.5Hz,2H),1.24(t,J=7.5Hz,3H).HRMS(ESI+)m / zcalculated for C 24 H 21 BrN9[M+H] + ,514.1098;found,514.1101.

[0232] 5. Synthesis of N-(2-bromo-4-fluorophenyl)-2-(3-ethyl-[1,2,4]triazolo[4,3-a]pyridin-6-yl)-6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-3-amine (J-55)

[0233] Intermediate N-(1-fluoro-3-bromo-4-benzene)formamide was prepared by replacing 4-fluoroaniline with 1 mmol of 1-fluoro-3-bromo-4-aniline according to step 4 of Example 2; intermediate N-(1-fluoro-3-bromo-4-benzene)formamide was prepared by replacing intermediate 1-fluoro-4-isocyanate with 0.4 mmol of intermediate 1-fluoro-3-bromo-4-isocyanate using the preparation method of Example 6, yielding 137.1 mg of a pale yellow solid with a yield of 45.2%. 1 HNMR(300MHz,DMSO-d6)δ9.16(s,1H),8.53(d,J=18.5Hz,2H),8.32(s,1H),8.03(d,J=8.4Hz,2H),7.97(d,J=9.6Hz,1H),7.81(d,J=9.8Hz,1H), 7.70-7.57(m,1H),6.92(t,J=8.3Hz,1H),6.22(dd,J=8.8,5.0Hz,1H),3.88(s,3H),3.00(q,J=7.5Hz,2H),1.27(t,J=7.8Hz,3H).HRMS(ESI+)m / z calculated for C 24 H20 BrFN9[M+H] + ,532.1004; found,532.1005.

[0234] Example 17 BRD4(BD1) protein inhibitory activity assay

[0235] The inhibitory activity of the compounds against BRD4 BD1 was tested using homogeneous time-resolved fluorescence (HTRF). Compounds 1–29 were diluted to 0.2 μM with DMSO, and compounds 30–56 were diluted to 1 μM with DMSO. The positive control (+)-JQ1 (BPS, Cat. No. 27402) was diluted to 1 μM and 0.2 μM with DMSO, and a blank control was added with DMSO. The compound solutions were transferred to different 384-well plates. One plate contained 0.2 μM of compound 1–29, 0.2 μM of the positive control, and blank control DMSO; the other plate contained 1 μM of compound 30–56, 1 μM of the positive control, and blank control DMSO. Two x BRD4(D1) (RBC, Cat. No. RD-11-157) were added to each plate, followed by two x compounds 1–56. After shaking for 30 seconds, the plates were incubated at room temperature for 1 hour. The HTRF signal values ​​were read using an EnVision detector (Ex at 340nm, Emat at 615nm & 665nm). Inhibition rate (%) = (Control wells - Experimental wells) / (Control wells - Blank wells) * 100. The results are shown in Table 1.

[0236] Table 1. Inhibition rate of compound J1-56 and positive control JQ1 against BRD4(BD1)

[0237]

[0238]

[0239] ND = Not determined.

[0240] The above data indicate that the compound has a significant inhibitory effect on BRD4(BD1) protein, and further research is warranted.

[0241] Example 18: In vitro antitumor cell proliferation assay of the compound:

[0242] Select healthy MV-4-11 and HCT116 cells, collect and centrifuge them, resuspend them in PBS (phosphate-buffered saline, pH 7.2, 10 mM), centrifuge at 1400 r / min using a benchtop low-speed centrifuge (Changsha Weierkang Xiangying Centrifuge Co., Ltd., TDZ4-WS), discard the supernatant, resuspend the cells in 1 mL of 10% serum culture medium, mix well, count, and dilute to a final cell concentration of 9000 cells / 100 μL. After mixing the cells, add them to a 96-well plate (100 μL per well), ensuring a uniform cell density in each well. Compounds 1–29 and the positive control (+)-JQ1 (BPS, Cat. No. 27402) were administered at concentrations of 10 μM and 1 μM, respectively (100 μL per well). A blank control (200 μL of 10% serum culture medium) and a negative control (100 μL of cell suspension) were also included. The plates were incubated at 37°C, 5% CO2. Three days after administration, under light-protected conditions, 20 μL of CCK-8 was added to each well, and the absorbance was measured using a ThermoFisher (51119670DP, wavelength 450 nm) to calculate the inhibition rate. Inhibition rate (%) = (OD value of negative control group - OD value of experimental group) / OD value of negative control group * 100%.

[0243] Based on the inhibition rate results, after treating MV-4-11 cells in good growth condition according to the above steps, the final cell concentration was 9000 cells / 100μL. Compounds 3, 6, 8, 9, 12-16 at different concentrations (20μM, 5μM, 1μM, 0.5μM, 0.25μM, 0.125μM, 0.0625μM, 0.03125μM) and positive control (+)-JQ1 (BPS, Cat. No. 27402) were administered (100μL per well). A blank control (200μL 10% serum culture medium) and a negative control (100μL cell suspension) were also set up and cultured in an incubator (37℃, 5% CO2). Three days after administration, 20 μL of CCK-8 was added to each well under light-protected conditions, and the absorbance was measured using a microplate reader (ThermoFisher, 51119670DP, wavelength 450 nM) to calculate the IC50.

[0244] Based on the inhibition rate results, HCT116 cells in good growth condition were treated according to the above steps to achieve a final cell concentration of 9000 cells / 100 μL. Compounds 5, 6, 9, 11, 15–17, and 22–24 at different concentrations (100 μM, 33.33 μM, 11.11 μM, 3.70 μM, 1.23 μM, 0.41 μM, 0.14 μM, 0.046 μM, 0.015 μM) and the positive control (+)-JQ1 (BPS, Cat. No. 27402) were administered (100 μL per well). A blank control (200 μL 10% serum culture medium) and a negative control (100 μL cell suspension) were also included. All cells were incubated in an incubator (37℃, 5% CO2). Three days after administration, 20 μL of CCK-8 was added to each well under light-protected conditions, and the absorbance was measured using a ThermoFisher (51119670DP, wavelength 450 nm) to calculate the IC50. The results are shown in Table 2.

[0245] Table 2. Results of the in vitro antiproliferative activity of compound J1-29 and positive control JQ1 on tumor cells.

[0246]

[0247]

[0248] ND = Not determined.

[0249] Select healthy MV4-11 and HL-6 cells and treat them according to the above steps to achieve a final cell concentration of 9000 / 100 μL. Mix the cells thoroughly and add them to a 96-well plate (100 μL per well), ensuring a consistent cell density in each well. Administer different concentrations (100 μM, 33.33 μM, 11.11 μM, 3.70 μM, 1.23 μM, 0.41 μM, 0.14 μM, 0.046 μM, 0.015 μM) of compounds 31, 32, 35, 38, 46, 52, and 55, along with the positive control (+)-JQ1 (BPS, Cat. No. 27402) (100 μL per well). A blank control (200 μL 10% serum culture medium) and a negative control (100 μL cell suspension) are also included. The plates are incubated at 37°C, 5% CO2. Three days after administration, 20 μL of CCK-8 was added to each well under light-protected conditions, and the absorbance was measured using a microplate reader (ThermoFisher, 51119670DP, wavelength 450 nM) to calculate the IC50.

[0250] Inhibition rate (%) = (OD value of negative control group - OD value of experimental group) / OD value of negative control group * 100%. IC50 values ​​were calculated using GraphPad Prism 7.0 software. The results are shown in Table 3.

[0251] Table 3 shows the in vitro antiproliferative activity of some compounds and the positive control JQ1 on tumor cells.

[0252] Compound numbering MV-4-11IC50(μM) HL-6IC50(μM) J-31 4.90±0.34 8.25±0.60 J-32 19.99±0.35 21.69±2.83 J-35 0.05±0.01 ND J-38 1.95±0.34 2.83±0.49 J-46 2.81±0.34 3.24±0.50 J-52 8.81±1.40 6.74±1.42 J-55 0.02±0.01 ND JQ1 3.05±0.01 3.15±0.21

[0253] ND = Not determined.

[0254] The above data show the antiproliferative activity of the synthesized compounds against tumor cells. The results indicate that at a concentration of 10 μM, all compounds exhibited significant antiproliferative activity (>70%) against MV-4-11 leukemia cells, with J-35 and J-55 showing better IC50 against MV-4-11 cells than the positive control drug JQ1. At a concentration of 10 μM, all compounds showed good antiproliferative activity against HCT116 and HL-6 cells, with slightly weaker antiproliferative activity at 1 μM.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is shown below: or .

2. A pharmaceutical composition, characterized in that, It comprises the compound of claim 1 or a pharmaceutically acceptable salt thereof.

3. The pharmaceutical composition according to claim 2, characterized in that, It also includes pharmaceutically acceptable carriers or excipients.

4. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2 or 3, in the preparation of an inhibitor having BRD4 inhibitory activity.

5. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2 or 3, in the preparation of a medicament for treating leukemia.

6. The application according to claim 5, characterized in that, The concentration of the compound is 1~10µM.

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