Pyrazole-imidazole-benzene compound as well as preparation method and application thereof

Abstract

The invention provides a pyrazole-imidazole-benzene compound as well as a preparation method and application thereof, and belongs to the fields of medicinal chemistry and pharmacotherapeutics. The compound shown in the formula I or the pharmaceutically acceptable salt thereof provided by the invention has good Aurora A kinase and JAK2 kinase inhibitory activity, can effectively inhibit the expression of Aurora A kinase and JAK2 kinase, also has an obvious inhibitory effect on human triple negative breast cancer cells MDA-MB-231, is higher than that of a positive control drug, can be applied to the aspect of preparing drugs for tumor-related diseases, and can be used for preparing drugs for treating tumor-related diseases. The invention particularly relates to application of the compound in the aspect of preparing double-target inhibitors of Aurora A kinase and JAK2 kinase. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry and pharmacotherapy, and specifically relates to a pyrazole-imidazole-benzene compound, a preparation method of the compound and a drug combination containing the compound. Background Art

[0002] Currently, breast cancer is one of the most common cancers in women and has long plagued women's health. Among breast cancers, triple negative breast cancer (TNBC) is the most challenging type, accounting for about 15-20% of all breast cancers. It is a difficult and key point in the treatment of breast cancer due to its poor prognosis, low tumor differentiation, high heterogeneity, strong invasiveness, high recurrence and metastasis rate, and limited treatment options.

[0003] Clinically, TNBC, characterized by negative estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (2HER2), is a more challenging type of breast cancer, accounting for about 15-20% of all breast cancers. Studies have shown that in the molecular targeted therapy of TNBC, due to the involvement of different proteins and enzymes and the complexity of the signal transduction system, after inhibiting one pathway, tumor-related signal molecules can also transmit information through other pathways by single-target kinase inhibitors, resulting in poor anti-tumor effects. Multi-target kinase inhibitors are one of the ways to solve these problems. They can simultaneously regulate all links that lead to the disease without affecting the relationship between the conduction systems and are not prone to drug resistance. The occurrence of TNBC is often accompanied by overexpression and proliferation of Aurora kinase family members, leading to abnormalities in the mitotic process and instability of the tumor genome. At the same time, JAK2 gene amplification or mutation is often detected in the somatic chromosome 20p24.1 region of TNBC patients. In addition, studies have shown that the synergistic effect of co-inhibiting Aurora kinase and JAK2 kinase has better therapeutic efficacy in tumor treatment. Summary of the invention

[0004] The purpose of the present invention is to provide a pyrazole-imidazole-benzene compound based on the prior art. The compound has good Aurora A kinase and JAK2 kinase inhibitory activity, can effectively inhibit the expression of Aurora A kinase and JAK2 kinase, and has a significant inhibitory effect on human triple-negative breast cancer cells MDA-MB-231, which is higher than the positive control drug, and can be used in the preparation of drugs for tumor-related diseases.

[0005] The second object of the present invention is to provide a pharmaceutical composition which uses the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient or the main active ingredient, supplemented with a pharmaceutically acceptable carrier.

[0006] The third object of the present invention is to provide a use of the above compound in the field of medicine.

[0007] The technical solution of the present invention is as follows:

[0008]

[0009] Among them,

[0010] R 1 represents X, a heteroalicyclic group or a substituted heteroalicyclic group, and the substituted heteroalicyclic group can be mono-substituted or multi-substituted by any of the following substituents: C1-C4 alkyl, C1-C4 alkoxy, hydroxy or halogen; X represents the following groups:

[0011]

[0012] R 3 represents C1-C4 alkyl, C1-C4 haloalkyl or phenyl;

[0013] R 4 represents C1-C4 alkyl, C1-C4 haloalkyl or phenyl;

[0014] R 2 represents C3-C10 cycloalkyl, substituted C3-C10 cycloalkyl, C1-C6 alkyl, phenyl or substituted phenyl; the substituted C3-C10 cycloalkyl or substituted phenyl can be mono-substituted or multi-substituted by any of the following substituents: C1-C4 alkyl, C1-C4 alkoxy, hydroxy or halogen.

[0015] In a preferred embodiment, R 1 represents X, a morpholine ring, a piperidine ring, a piperazine ring, a pyrrolidine ring, a substituted morpholine ring, a substituted piperidine ring or a substituted piperazine ring, and the substituted morpholine ring, substituted piperidine ring or substituted piperazine ring can be mono-substituted or multi-substituted by any of the following substituents: methyl, ethyl, methoxy or ethoxy; R 3 represents methyl or ethyl; R 4 represents methyl or ethyl.

[0016] In a more preferred embodiment, R 1 represents X, a morpholine ring, a piperidine ring, a piperazine ring, a substituted piperidine ring or a substituted piperazine ring, and the substituted piperidine ring or substituted piperazine ring can be mono-substituted or multi-substituted by any of the following substituents: methyl, ethyl.

[0017] In a particularly preferred embodiment, R 1 represents the following groups:

[0018]

[0019] In a preferred embodiment, R 2 represents cyclopropane, cyclobutane, cyclopentane, substituted cyclopentane, cyclohexane, substituted cyclopentane, cycloheptane, propyl, isopropyl, tert-butyl, phenyl, substituted phenyl; the substituted cyclopentane, substituted cyclopentane or substituted phenyl may be mono-substituted or multi-substituted by any of the following substituents: methyl, ethyl, methoxy, ethoxy, fluorine or chlorine.

[0020] In a more preferred embodiment, R 2 represents cyclopropane, cyclobutane, cyclopentane, cyclohexane, substituted cyclopentane, cycloheptane, propyl, isopropyl, tert-butyl, phenyl, substituted phenyl; the substituted cyclopentane, substituted cyclopentane or substituted phenyl may be mono-substituted or multi-substituted by any of the following substituents: methyl, methoxy or fluorine.

[0021] In a particularly preferred embodiment, R 2 represents the following groups:

[0022]

[0023] In an even more particularly preferred embodiment, R 1 represents the following groups:

[0024]

[0025] R 2 represents the following groups:

[0026]

[0027] Furthermore, in the compound of formula I or a pharmaceutically acceptable salt thereof, the compound is selected from the following compounds:

[0028]

[0029]

[0030]

[0031]

[0032] The present invention provides a pharmaceutical composition which uses the compound of the present invention or a pharmaceutically acceptable salt thereof as the active ingredient or the main active ingredient, supplemented with a pharmaceutically acceptable carrier.

[0033] The compounds of the present invention or their pharmaceutically acceptable salts can be used in the preparation of drugs for treating diseases related to tumors, for example, drugs that are dual-target inhibitors of Aurora A kinase and JAK2 kinase.

[0034] Unless otherwise specified, the following terms used in the specification and claims have the meanings discussed below:

[0035] "Pharmaceutically acceptable salts" refer to those salts that retain the biological activity and properties of the parent compound. Such salts include:

[0036] (1) Salts formed by reacting the free base of the parent compound with inorganic acids or organic acids. Inorganic acids include hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid, perchloric acid, etc.; organic acids include acetic acid, trifluoroacetic acid, propionic acid, acrylic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, oxalic acid, (D)- or (L)-malic acid, fumaric acid, maleic acid, ascorbic acid, camphoric acid, benzoic acid, hydroxybenzoic acid, γ-hydroxybutyric acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, cinnamic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, aspartic acid, stearic acid, mandelic acid, succinic acid, glutaric acid, malonic acid, etc.

[0037] (2) Salts formed by replacing the acidic protons in the parent compound with metal ions or coordinating with organic bases. Metal ions such as alkali metal ions, alkaline earth metal ions or aluminum ions; organic bases such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, quinine, etc.

[0038] "Pharmaceutical composition" refers to a mixture of one or more of the compounds of the present invention or their pharmaceutically acceptable salts, solvates, hydrates or prodrugs with other chemical components, such as pharmaceutically acceptable carriers. The purpose of the pharmaceutical composition is to facilitate the administration process to animals.

[0039] "Pharmaceutical carrier" or "pharmaceutically acceptable carrier" refers to the inactive components in a pharmaceutical composition that do not cause significant irritation to the organism and do not interfere with the biological activity and properties of the administered compound, such as but not limited to: calcium carbonate, calcium phosphate, various sugars (such as lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic polymers or methacrylic polymers, gels, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

[0040] "Alkyl" means a saturated aliphatic hydrocarbon group having 1 to 20 carbon atoms, including straight-chain and branched-chain groups (the numerical ranges mentioned in this application, such as "1-20", refer to the group, which is an alkyl group at this time and can contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms). More preferably, the alkyl group is a medium-sized alkyl group having 1 to 10 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl, etc. Preferably, the alkyl group is a lower alkyl group having 1 to 8 or 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl or tert-butyl, etc. The alkyl group can be substituted or unsubstituted. When it is a substituted alkyl group, the substituent is preferably one or more, more preferably 1 to 3, and most preferably 1 or 2 substituents.

[0041] "Aromatic heterocycle" or "heteroaryl" means a monocyclic or fused-ring group having 5 to 12 ring atoms, containing one, two, three or four ring heteroatoms selected from N, O or S, and the remaining ring atoms are C, and further having a completely conjugated π-electron system. Non-limiting examples of unsubstituted heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine and carbazole. The heteroaryl group can be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one, two or three, and even more preferably one or two, independently selected from the following groups, including: lower alkyl, trihaloalkyl, halogen, hydroxy, lower alkoxy, mercapto, (lower alkyl)thio, cyano, acyl, thioacyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, nitro, N-sulfonamido, S-sulfonamido. Preferred heteroaryl groups are optionally substituted with one or two substituents, and the substituents are independently selected from halogen, lower alkyl, trihaloalkyl, hydroxy, mercapto, cyano, N-acylamino, mono- or dialkylamino, carboxyl or N-sulfonamido.

[0042] "Heteroalicyclic group" means a monocyclic or fused-ring group having 5 to 9 ring atoms in the ring, wherein one or two ring atoms are heteroatoms selected from N, O or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are C. These rings can have one or more double bonds, but these rings do not have a completely conjugated π-electron system. Non-limiting examples of unsubstituted heteroalicyclic groups are pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, etc.

[0043] "Hydroxy" means the -OH group.

[0044] "Alkoxy" means -O-(unsubstituted alkyl) and -O-(unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0045] "Halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

[0046] Adopting the technical solution of the present invention, the advantages are as follows:

[0047] The pyrazole-imidazole-benzene compounds provided by the present invention have good inhibitory activities against Aurora A kinase and JAK2 kinase, can effectively inhibit the expression of Aurora A kinase and JAK2 kinase, and also have obvious inhibitory effects on human triple-negative breast cancer cells MDA-MB-231, which are higher than those of the positive control drug, and can be applied to the preparation of drugs for diseases related to tumors. Brief Description of the Drawings

[0048] Figure 1 Shows the effects of compounds ZT-1B, ZT-2F and ZT-2M on the expression of Aurora A and STAT3 proteins;

[0049] Figure 2 Shows the cell cycle arrest of compound ZT-2F-induced MDA-MB-231 cells in the G2 phase;

[0050] Figure 3 Shows the apoptosis experiment of compound ZT-2F-induced MDA-MB-231 cells. Detailed Description of the Invention

[0051] According to the following examples, the present invention can be better understood. However, those skilled in the art can easily understand that the content described in the examples is only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0052] I. Synthetic route of ZT series compounds in the present invention:

[0053] Using 4-nitropyrazole-3-carboxylic acid and 4-methylacetophenone as starting materials, through reaction steps such as N-alkylation, α-bromination of 4-methylacetophenone, esterification, imidazole ring formation, nitro reduction, urea formation reaction, etc., the ZT series compounds are finally obtained. The specific synthetic steps are as follows:

[0054]

[0055] (1) Synthesis of Compound 1

[0056]

[0057] At room temperature, 4-nitropyrazole-3-carboxylic acid (2.5 g, 15.9 mmol) was dissolved in 25 mL of tetrahydrofuran (THF). Under stirring, 3,4-dihydro-2H-pyran (DHP) (4.02 g, 47.75 mmol) and p-toluenesulfonic acid monohydrate (PTSA) (0.15 g, 0.075 mmol) were added successively. Stirring was continued overnight at room temperature. After the reaction was complete as detected by TLC, the solvent in the reaction solution was concentrated under reduced pressure, and then 50 mL of EA was added for dissolution and extraction with 40 mL of water. The organic layer was collected, and the aqueous layer was further extracted with ethyl acetate (50 mL × 3). The combined organic phases were extracted with saturated NaCl aqueous solution three times. The organic layer was dried over anhydrous sodium sulfate, filtered by suction, and a dark brown solution was obtained, which was directly triturated. Purification by silica gel column chromatography (dichloromethane:methanol = 200:1, V / V) gave 3.65 g of the target product with a yield of 95.3%.

[0058] (2) Synthesis of Compound 2

[0059]

[0060] At room temperature, p-methylacetophenone (20 mL, 150 mmol) was dissolved in 160 mL of acetonitrile solution. Subsequently, NBS (29.2 g, 164 mmol) and AIBN (2.46 g, 14.98 mmol) were added successively. After addition, the temperature of the reaction solution was raised to 85 °C and heated under reflux for 8 h. The reaction was complete as detected by TLC, and a yellow reaction solution was obtained. The acetonitrile was removed by concentration under reduced pressure to give a yellow oily compound, which was dissolved in 100 mL of CH 2 Cl 2 and extracted with 200 mL of water. The organic layer was collected. The aqueous layer was further extracted with CH 2 Cl 2 (50 mL × 3). The organic layer was collected. Then the organic layer was extracted successively with 1 M dilute hydrochloric acid solution, saturated sodium bicarbonate aqueous solution, and saturated NaCl solution. The organic layer was dried over anhydrous sodium sulfate, filtered by suction, concentrated under reduced pressure to give a yellow oil, which was dissolved in 20 mL of ethyl acetate and heated with stirring at 78 °C until the compound was completely dissolved. 200 mL of petroleum ether was quickly added during heating, and then the reaction solution was carefully placed in the refrigerator and left to stand overnight. A large amount of white solid precipitated. 30.2 g of the target product was obtained with a yield of 95%.

[0061] (3) Synthesis of Compound 3a

[0062]

[0063] At room temperature, compound 2 (10 g, 47 mmol) was dissolved in 60 mL of acetonitrile solution. While stirring, potassium carbonate (6.48 g, 47 mmol), a catalytic amount of potassium iodide, and morpholine (4.08 g, 47 mmol) were added successively. After the addition was complete, the temperature of the reaction solution was raised to 85 °C and heated under reflux for 8 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then lowered to room temperature, and the mixture was filtered by suction. The filter cake was washed thoroughly with ethyl acetate to obtain a dark yellow solution, which was directly triturated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 15 / 1, 12 / 1, 10 / 1, 8 / 1, V / V) to obtain 9.3 g of the target compound with a yield of 90%. 1 H NMR (400 MHz, Chloroform-d) δ 7.94 - 7.88 (m, 2H), 7.44 (d, J = 8.0 Hz, 2H), 3.76 - 3.67 (m, 4H), 3.55 (s, 2H), 2.60 (s, 3H), 2.45 (t, J = 4.6 Hz, 4H).

[0064] (4) Synthesis of compound 3b

[0065]

[0066] At room temperature, compound 2 (10 g, 47 mmol) was dissolved in 60 mL of acetonitrile solution. While stirring, potassium carbonate (6.48 g, 47 mmol), a catalytic amount of potassium iodide, and piperidine (3.83 g, 47 mmol) were added successively. After the addition was complete, the temperature of the reaction solution was raised to 85 °C and heated under reflux for 8 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then lowered to room temperature, and the mixture was filtered by suction. The filter cake was washed thoroughly with ethyl acetate to obtain a dark yellow solution, which was directly triturated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 8 / 1, 4 / 1, 1 / 1, V / V) to obtain 8.97 g of the target compound with a yield of 90%. 1 HNMR (400 MHz, Chloroform-d) δ 7.95 - 7.86 (m, 2H), 7.46 - 7.37 (m, 2H), 3.52 (s, 2H), 2.60 (s, 3H), 2.38 (s, 4H), 1.64 - 1.52 (m, 4H), 1.51 - 1.39 (m, 2H).

[0067] (5) Synthesis of compound 3d

[0068]

[0069] At room temperature, dissolve compound 2 (10 g, 47 mmol) in 60 mL of acetonitrile solution. While stirring, sequentially add potassium carbonate (6.48 g, 47 mmol), a catalytic amount of potassium iodide, and diethylamine (3.29 g, 47 mmol). After addition, raise the temperature of the reaction solution to 85 °C and heat under reflux for 8 h. Detect the completion of the reaction by TLC. Cool the temperature of the reaction solution to room temperature, perform suction filtration, wash the filter cake with ethyl acetate until clean to obtain a dark yellow solution. Directly prepare sand, and separate and purify by silica gel column chromatography (petroleum ether:ethyl acetate = 8 / 1, 4 / 1, 1 / 1, V / V) to obtain 9.3 g of the target compound with a yield of 88.9%. 1 H NMR (400 MHz, Chloroform-d) δ 7.91 (dd, J = 7.6, 2.4 Hz, 2H), 7.46 (d, J = 7.6 Hz, 2H), 3.63 (s, 2H), 2.57 (dd, J = 21.2, 5.2 Hz, 7H), 1.11 - 1.01 (m, 6H).

[0070] (6) Synthesis of compound 3e

[0071]

[0072] At room temperature, dissolve compound 2 (10 g, 47 mmol) in 60 mL of acetonitrile solution. While stirring, sequentially add potassium carbonate (6.48 g, 47 mmol), a catalytic amount of potassium iodide, and N-methylpiperazine (4.7 g, 47 mmol). After addition, raise the temperature of the reaction solution to 85 °C and heat under reflux for 8 h. Detect the completion of the reaction by TLC. Cool the temperature of the reaction solution to room temperature, perform suction filtration, wash the filter cake with ethyl acetate until clean to obtain a dark yellow solution. Directly prepare sand, and separate and purify by silica gel column chromatography (petroleum ether:ethyl acetate = 4 / 1, 1 / 1, V / V) to obtain 8.24 g of the target compound with a yield of 76%.

[0073] (7) Synthesis of compound 3f

[0074]

[0075] At room temperature, dissolve compound 2 (10 g, 47 mmol) in 60 mL of acetonitrile solution. While stirring, sequentially add potassium carbonate (6.48 g, 47 mmol), a catalytic amount of potassium iodide, and 3-methylpiperidine (4.66 g, 47 mmol). After addition, raise the temperature of the reaction solution to 85 °C and heat under reflux for 8 h. Detect the completion of the reaction by TLC. Cool the temperature of the reaction solution to room temperature, perform suction filtration, wash the filter cake with ethyl acetate until clean to obtain a dark yellow solution. Directly prepare sand, and separate and purify by silica gel column chromatography (petroleum ether:ethyl acetate = 15 / 1, 12 / 1, 10 / 1, 8 / 1, 4 / 1, 1 / 1, V / V) to obtain 8.57 g of the target compound with a yield of 78.8%.

[0076] (8) Synthesis of Compound 4a

[0077]

[0078] At 0 °C, dissolve compound 3a (1.383 g, 6.3 mmol) in 50 mL of 48% HBr solution and stir for 5 min. Then, dropwise add a solution of bromine (1.20 g, 150 mmol) dissolved in 3 mL of 48% HBr into the above solution at a dropping rate of 25 seconds. After the addition is complete, remove the ice bath and stir the reaction solution at room temperature for 2 - 3 h. When the reaction is complete as detected by TLC, adjust the pH of the reaction solution to neutral with saturated sodium bicarbonate solution, extract with 30 mL of ethyl acetate, extract the aqueous layer with 30 mL of ethyl acetate three times, and combine the organic layers. Extract the organic layer three times with saturated NaCl solution, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure to remove ethyl acetate, and directly and rapidly add it to the next step without purification, with a yield of 90%.

[0079] (9) Synthesis of Compound 4b

[0080]

[0081] At 0 °C, dissolve compound 3b (1.74 g, 8.0 mmol) in 50 mL of 48% HBr solution and stir for 5 min. Then, dropwise add a solution of bromine (1.20 g, 15 mmol) dissolved in 5 mL of 48% HBr into the above solution at a dropping rate of 25 seconds. After the addition is complete, remove the ice bath and stir the reaction solution at room temperature for 2 - 3 h. When the reaction is complete as detected by TLC, adjust the pH of the reaction solution to neutral with saturated sodium bicarbonate solution. Extract with 30 mL of ethyl acetate, extract the aqueous layer with 30 mL of ethyl acetate three times, and combine the organic layers. Extract the organic layer three times with saturated NaCl solution, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure to remove ethyl acetate, and directly and rapidly add it to the next step without purification, with a yield of 90%.

[0082] (10) Synthesis of Compound 4d

[0083]

[0084] At 0 °C, compound 3d (1.80 g, 8.7 mmol) was dissolved in 50 mL of 48% HBr solution and stirred for 5 min. Then, a solution of bromine (2.11 g, 13 mmol) dissolved in 5 mL of 48% HBr was added dropwise to the above solution at a dropping rate of 25 seconds. After the addition was complete, the ice bath was removed, and the reaction mixture was stirred at room temperature for 2 - 3 h. When the reaction was completed as detected by TLC, the pH of the reaction mixture was adjusted to neutral with saturated sodium bicarbonate solution. It was extracted with 30 mL of ethyl acetate, and the aqueous layer was extracted with 30 mL of ethyl acetate three times. The organic layers were combined. The organic layer was extracted three times with saturated NaCl solution, the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove ethyl acetate, and directly used in the next step without purification, with a yield of 90%.

[0085] (11) Synthesis of compound 4e

[0086]

[0087] At 0 °C, compound 3b (1.60 g, 6.9 mmol) was dissolved in 50 mL of 48% HBr solution and stirred for 5 min. Then, a solution of bromine (1.65 g, 10.3 mmol) dissolved in 5 mL of 48% HBr was added dropwise to the above solution at a dropping rate of 25 seconds. After the addition was complete, the ice bath was removed, and the reaction mixture was stirred at room temperature for 2 - 3 h. When the reaction was completed as detected by TLC, the pH of the reaction mixture was adjusted to neutral with saturated sodium bicarbonate solution. It was extracted with 30 mL of ethyl acetate, and the aqueous layer was extracted with 30 mL of ethyl acetate three times. The organic layers were combined. The organic layer was extracted three times with saturated NaCl solution, the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove ethyl acetate, and directly used in the next step without purification, with a yield of 90%.

[0088] (12) Synthesis of compound 4f

[0089]

[0090] At 0 °C, compound 3f (1.383 g, 6.3 mmol) was dissolved in 50 mL of 48% HBr solution and stirred for 5 min. Then, a solution of bromine (1.20 g, 150 mmol) dissolved in 5 mL of 48% HBr was added dropwise to the above solution at a dropping rate of 25 seconds. After the addition was complete, the ice bath was removed, and the reaction mixture was stirred at room temperature for 2 - 3 h. When the reaction was completed as detected by TLC, the pH of the reaction mixture was adjusted to neutral with saturated sodium bicarbonate solution. It was extracted with 30 mL of ethyl acetate, and the aqueous layer was extracted with 30 mL of ethyl acetate three times. The organic layers were combined. The organic layer was extracted three times with saturated NaCl solution, the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove ethyl acetate, and directly used in the next step without purification, with a yield of 90%.

[0091] (13) Synthesis of compound 5a

[0092]

[0093] At room temperature, compound 1 (4.82 g, 20 mmol) was dissolved in acetonitrile solution (100 mL), stirred until completely dissolved, and then potassium carbonate (2.76 g, 20 mmol), a catalytic amount of potassium iodide, and compound 4a (7.56 g, 20 mmol) dissolved in 50 mL of acetonitrile solution were added to the above solution in sequence. The mixture was continuously stirred at room temperature overnight. After the reaction was detected to be complete by TLC, it was filtered by suction, the filter cake was washed clean with ethyl acetate, and the filtrate was directly made into sand. The target product was obtained as a pale yellow solid by silica gel column chromatography (petroleum ether:ethyl acetate = 6 / 1, 3 / 1, 1 / 1, 1 / 2, V / V) with a yield of 68%.

[0094] (14) Synthesis of compound 5b

[0095]

[0096] At room temperature, compound 1 (4.82 g, 20 mmol) was dissolved in acetonitrile solution (100 mL), stirred until completely dissolved, and then potassium carbonate (2.76 g, 20 mmol), a catalytic amount of potassium iodide, and compound 4b (7.56 g, 20 mmol) dissolved in 50 mL of acetonitrile solution were added to the above solution in sequence. The mixture was continuously stirred at room temperature overnight. After the reaction was detected to be complete by TLC, the reaction solution was filtered by suction, the filter cake was washed clean with ethyl acetate, and the filtrate was directly made into sand. The target product was obtained as a pale yellow solid by silica gel column chromatography (petroleum ether:ethyl acetate = 6 / 1, 3 / 1, 1 / 1, V / V) with a yield of 73%.

[0097] (15) Synthesis of compound 5d

[0098]

[0099] At room temperature, compound 1 (4.82 g, 20 mmol) was dissolved in acetonitrile solution (100 mL), stirred until completely dissolved, and then potassium carbonate (2.76 g, 20 mmol), a catalytic amount of potassium iodide, and compound 4d (7.56 g, 20 mmol) dissolved in 50 mL of acetonitrile solution were added to the above solution in sequence. The mixture was continuously stirred at room temperature overnight. After the reaction was detected to be complete by TLC, it was filtered by suction, the filter cake was washed clean with ethyl acetate, and the filtrate was directly made into sand. The target product was obtained as a pale yellow solid by silica gel column chromatography (petroleum ether:ethyl acetate = 6 / 1, 3 / 1, 1 / 1, V / V) with a yield of 63%. 11H NMR (400 MHz, Chloroform-d) δ 8.45 (s, 1H), 7.98 (s, 2H), 7.79 (s, 2H), 5.62 (s, 2H), 5.50 (d, J = 9.0 Hz, 1H), 4.12 (d, J = 11.2 Hz, 2H), 3.76 (q, J = 10.0, 7.3 Hz, 2H), 2.95 (s, 4H), 2.32 - 2.21 (m, 2H), 2.02 (s, 2H), 1.71 (t, J = 5.6 Hz, 2H), 1.35 (s, 6H).

[0100] (16) Synthesis of Compound 5e

[0101]

[0102] At room temperature, compound 1 (4.82 g, 20 mmol) was dissolved in acetonitrile solution (100 mL), stirred until completely dissolved, and then potassium carbonate (2.76 g, 20 mmol), a catalytic amount of potassium iodide, and compound 4e (7.56 g, 20 mmol) dissolved in 50 mL of acetonitrile solution were added to the above solution in sequence. Stirring was continued at room temperature overnight. After TLC detection showed that the reaction was complete, filtration was carried out by suction, the filter cake was washed clean with ethyl acetate, and the filtrate was directly made into sand. The target product was obtained as a pale yellow solid after separation and purification by silica gel column chromatography (petroleum ether:ethyl acetate = 4 / 1, 1 / 1, 1 / 2, V / V), with a yield of 32%.

[0103] (17) Synthesis of Compound 5f

[0104]

[0105] At room temperature, compound 1 (4.82 g, 20 mmol) was dissolved in acetonitrile solution (100 mL), stirred until completely dissolved, and then potassium carbonate (2.76 g, 20 mmol), a catalytic amount of potassium iodide, and compound 4f (7.56 g, 20 mmol) dissolved in 50 mL of acetonitrile solution were added to the above solution in sequence. Stirring was continued at room temperature overnight. After TLC detection showed that the reaction was complete, filtration was carried out by suction, the filter cake was washed clean with ethyl acetate, and the filtrate was directly made into sand. The target product was obtained as a pale yellow solid after separation and purification by silica gel column chromatography (petroleum ether:ethyl acetate = 6 / 1, 3 / 1, 1 / 1, 1 / 2, V / V), with a yield of 65%.

[0106] (18) Synthesis of Compound 6a

[0107]

[0108] At room temperature, dissolve compound 5a (2.29 g, 5 mmol) in 20 mL of acetic acid solution. With stirring, add ammonium acetate (4.62 g, 60 mmol). After addition, raise the temperature of the reaction solution to 130 °C and react for 5 h. Monitor the reaction by TLC until completion. Cool the reaction solution to room temperature, dilute it with ethanol, and remove acetic acid by concentration under reduced pressure to obtain a dark yellowish-brown oily substance. At 0 °C, slowly add the reaction solution dropwise to a saturated sodium bicarbonate solution on ice, adjust the pH of the reaction solution to neutral, and a large amount of yellow solid will precipitate. Stir for 1 - 2 h, filter by suction, and dry the filter cake in a vacuum drying oven. Without purification, it can be directly used for the next reaction to obtain 0.81 g of the target product with a yield of 45%.

[0109] (19) Synthesis of compound 6b

[0110]

[0111] At room temperature, dissolve compound 5b (2.28 g, 5 mmol) in 20 mL of acetic acid solution. With stirring, add ammonium acetate (4.62 g, 60 mmol). After addition, raise the temperature of the reaction solution to 130 °C and react for 5 h. Monitor the reaction by TLC until completion. Cool the reaction solution to room temperature, dilute it with ethanol, and remove acetic acid by concentration under reduced pressure to obtain a dark yellowish-brown oily substance. At 0 °C, slowly add the reaction solution dropwise to a saturated sodium bicarbonate solution on ice, adjust the pH of the reaction solution to neutral, and a large amount of yellow solid will precipitate. Stir for 1 - 2 h, filter by suction, and dry the filter cake in a vacuum drying oven. Without purification, it can be directly used for the next reaction to obtain 0.90 g of the target product with a yield of 50%.

[0112] (20) Synthesis of compound 6d

[0113]

[0114] At room temperature, dissolve compound 5d (2.20 g, 5 mmol) in 20 mL of acetic acid solution. With stirring, add ammonium acetate (4.62 g, 60 mmol). After addition, raise the temperature of the reaction solution to 130 °C and react for 5 h. Monitor the reaction by TLC until completion. Cool the reaction solution to room temperature, dilute it with ethanol, and remove acetic acid by concentration under reduced pressure to obtain a dark yellowish-brown oily substance. At 0 °C, slowly add the reaction solution dropwise to a saturated sodium bicarbonate solution on ice, adjust the pH of the reaction solution to neutral, and a large amount of yellow solid will precipitate. Stir for 1 - 2 h, filter by suction, and dry the filter cake in a vacuum drying oven. Without purification, it can be directly used for the next reaction to obtain 0.87 g of the target product with a yield of 43%.

[0115] (21) Synthesis of compound 6e

[0116]

[0117] At room temperature, dissolve compound 5e (2.29 g, 5 mmol) in 20 mL of acetic acid solution. With stirring, add ammonium acetate (4.62 g, 60 mmol). After addition, raise the temperature of the reaction solution to 130 °C and react for 5 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, dilute the reaction solution with ethanol, and remove acetic acid by concentration under reduced pressure to obtain a dark brownish-yellow oily substance. At 0 °C, slowly add the reaction solution dropwise to ice-cold saturated sodium bicarbonate solution, adjust the pH of the reaction solution to neutral, and a large amount of yellow solid precipitates. Stir for 1 - 2 h, filter by suction, and dry the filter cake in a vacuum drying oven. Without purification, it can be directly used for the next reaction to obtain 0.24 g of the target product with a yield of 30%.

[0118] (22) Synthesis of compound 6f

[0119]

[0120] At room temperature, dissolve compound 5f (2.35 g, 5 mmol) in 20 mL of acetic acid solution. With stirring, add ammonium acetate (4.62 g, 60 mmol). After addition, raise the temperature of the reaction solution to 130 °C and react for 5 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, dilute the reaction solution with ethanol, and remove acetic acid by concentration under reduced pressure to obtain a dark brownish-yellow oily substance. At 0 °C, slowly add the reaction solution dropwise to ice-cold saturated sodium bicarbonate solution, adjust the pH of the reaction solution to neutral, and a large amount of yellow solid precipitates. Stir for 1 - 2 h, filter by suction, and dry the filter cake in a vacuum drying oven. Without purification, it can be directly used for the next reaction to obtain 0.91 g of the target product with a yield of 50%.

[0121] (23) Synthesis of compound 7a

[0122]

[0123] At room temperature, dissolve compound 6a (1.77 g, 5 mmol) in 100 mL of methanol solution, add 177 mg of palladium carbon (wetted with ca. 55% Water), charge with hydrogen, and then raise the temperature of the reaction solution to 45 °C and stir for 6 - 7 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, filter by suction, wash the filter cake thoroughly with methanol, concentrate the filtrate under reduced pressure to obtain 1.55 g of a dark brownish-yellow oily substance. Without purification, immediately use it in the next step with a yield of 85%.

[0124] (24) Synthesis of compound 7b

[0125]

[0126] At room temperature, dissolve compound 6b (1.76 g, 5 mmol) in 100 mL of methanol solution, add 176 mg of palladium on carbon (wetted with ca. 55% Water), charge with hydrogen, then raise the temperature of the reaction solution to 45 °C and stir for 6 - 7 h. Detect that the reaction is complete by TLC. Cool the reaction solution to room temperature, filter by suction, wash the filter cake thoroughly with methanol, concentrate the filtrate under reduced pressure to obtain 1.56 g of a yellowish-brown oily substance. Without purification, immediately use it in the next step, with a yield of 85.7%.

[0127] (25) Synthesis of compound 7d

[0128]

[0129] At room temperature, dissolve compound 6d (1.72 g, 5 mmol) in 100 mL of methanol solution, add 172 mg of palladium on carbon (wetted with ca. 55% Water), charge with hydrogen, then raise the temperature of the reaction solution to 45 °C and stir for 6 - 7 h. Detect that the reaction is complete by TLC. Cool the reaction solution to room temperature, filter by suction, wash the filter cake thoroughly with methanol, concentrate the filtrate under reduced pressure to obtain 1.54 g of a yellowish-brown oily substance. Without purification, immediately use it in the next step, with a yield of 85%.

[0130] (26) Synthesis of compound 7e

[0131]

[0132] At room temperature, dissolve compound 6e (1.84 g, 5 mmol) in 100 mL of methanol solution, add 184 mg of palladium on carbon (wetted with ca. 55% Water), charge with hydrogen, then raise the temperature of the reaction solution to 45 °C and stir for 6 - 7 h. Detect that the reaction is complete by TLC. Cool the reaction solution to room temperature, filter by suction, wash the filter cake thoroughly with methanol, concentrate the filtrate under reduced pressure to obtain 1.58 g of a yellowish-brown oily substance. Without purification, immediately use it in the next step, with a yield of 85.9%.

[0133] (27) Synthesis of compound 7f

[0134]

[0135] At room temperature, dissolve compound 6f (1.83 g, 5 mmol) in 100 mL of methanol solution, add 183 mg of palladium on carbon (wetted with ca. 55% Water), charge with hydrogen, then raise the temperature of the reaction solution to 45 °C and stir for 6 - 7 h. Detect that the reaction is complete by TLC. Cool the reaction solution to room temperature, filter by suction, wash the filter cake thoroughly with methanol, concentrate the filtrate under reduced pressure to obtain 1.63 g of a yellowish-brown oily substance. Without purification, immediately use it in the next step, with a yield of 83%.

[0136] (28) Synthesis of Compound ZT-1A

[0137]

[0138] At room temperature, dissolve compound 7a (300 mg, 0.92 mmol) in 20 mL of THF solution, then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and concentrate the solvent under reduced pressure. Without separation and purification, directly and rapidly add it to the next reaction. Dissolve compound [7a] in 15 mL of DMF solution, then add cyclohexylamine (0.64 g, 6.44 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h, then filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 of EA. Combine the organic layers. Directly form sand from the organic layer and perform column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V). 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.94 (d, J = 4.4 Hz, 1H), 7.90 - 7.75 (dd, J = 50.8, 7.8 Hz, 2H), 7.56 (s, 1H), 7.28 - 7.24 (t, J = 6.6 Hz, 2H), 7.06 (s, 1H), 3.54 (t, J = 4.4 Hz, 4H), 3.43 (s, 2H), 3.41 (s, 1H), 2.32 (s, 4H), 1.82 (d, J = 11.8 Hz, 2H), 1.73 - 1.41 (m, 3H), 1.31 - 1.03 (m, 5H).

[0139] (29) Synthesis of Compound ZT-1B

[0140]

[0141] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then cyclopentylamine (0.56 g, 6.44 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated and chromatographed on a basic alumina column (methylene chloride: methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.76 (s, 1H), 7.99 (s, 1H), 7.91 - 7.79 (dd, J = 40.1, 8.0 Hz, 2H), 7.61 (d, J = 2.0 Hz, 1H), 7.28 - 7.30 (d, J = 8.0 Hz, 2H), 7.15 (s, 1H), 3.96 (dt, J = 13.4, 6.8 Hz, 1H), 3.58 (t, J = 4.6 Hz, 4H), 3.46 (d, J = 4.9 Hz, 2H), 2.36 (s, 4H), 1.86 (dq, J = 44.6, 6.6, 6.2 Hz, 2H), 1.73 - 1.60 (m, 2H), 1.60 - 1.38 (m, 4H).

[0142] (30) Synthesis of compound ZT-1C

[0143]

[0144] At room temperature, dissolve compound 7a (300 mg, 0.92 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and concentrate the solvent under reduced pressure. Without separation and purification, directly and rapidly add it to the next reaction. Dissolve compound [7a] in 15 mL of DMF solution, and then add cyclopropylamine (0.58 g, 6.44 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h, and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 of EA. Combine the organic layers. Directly form sand from the organic layer, and perform column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ8.01(s,1H),7.81-7.79(d,J=8.0Hz,2H),7.61(d,J=2.4Hz,1H),7.27(d,J=8.0Hz,2H),7.19(s,1H),3.57(t,J=4.6Hz,4H),3.45(s,2H),2.70 -2.60(m,1H),2.35(s,4H),0.74(d,J=4.8Hz,2H),0.53-0.48(m,2H).

[0145] (31) Synthesis of compound ZT-1F

[0146]

[0147] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly used in the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then 2-methylcyclohexylamine (0.73 g, 6.44 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V) was carried out. 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.71 (s, 1H), 7.98 (s, 1H), 7.94 - 7.77 (m, 2H), 7.60 (s, 1H), 7.30 (d, J = 8.2 Hz, 2H), 6.99 (s, 1H), 3.58 (t, J = 4.6 Hz, 4H), 3.47 (s, 2H), 3.14 (s, 1H), 2.36 (s, 4H), 1.85 (d, J = 10.4 Hz, 1H), 1.75 - 1.14 (m, 9H), 0.96 - 0.85 (m, 3H).

[0148] (32) Synthesis of compound ZT-1K

[0149]

[0150] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution. Subsequently, CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly introduced into the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then 4-fluoroaniline (0.72 g, 6.44 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powdered compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powdered solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V) was carried out. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.80 - 9.75 (d, J = 21.8 Hz, 1H), 9.21 - 9.07 (d, J = 56.8 Hz, 1H), 8.08 - 8.07 (d, J = 6.8 Hz, 1H), 7.94 - 7.81 (dd, J = 43.2, 8.0 Hz, 2H), 7.64 - 7.63 (d, J = 2.0 Hz, 1H), 7.56 - 7.50 (m, 2H), 7.33 - 7.29 (t, J = 7.8 Hz, 2H), 7.16 - 7.09 (q, J = 8.6 Hz, 2H), 3.60 - 3.57 (t, J = 4.6 Hz, 4H), 3.47 - 3.46 (d, J = 4.4 Hz, 2H), 2.37 (s, 4H).

[0151] (33) Synthesis of compound ZT-1M

[0152]

[0153] At room temperature, dissolve compound 7a (300 mg, 0.92 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Detect that the reaction is complete by TLC. Cool the temperature of the reaction solution to room temperature, and then concentrate the solvent under reduced pressure. Without separation and purification, directly and quickly put it into the next reaction. Dissolve compound [7a] in 15 mL of DMF solution, and then add 4-methylcyclohexylamine (0.73 g, 6.44 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Detect that the reaction is complete by TLC. Cool the temperature of the reaction solution to room temperature, and then pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly grind the organic layer into sand and perform column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.65 (s, 1H), 7.95 - 7.93 (t, J = 4.6 Hz, 1H), 7.91 - 7.73 (m, 2H), 7.56 (s, 1H), 7.28 - 7.24 (t, J = 7.0 Hz, 2H), 7.05 (s, 1H), 3.65 (s, 1H), 3.55 - 3.53 (t, J = 4.6 Hz, 4H), 3.43 - 3.41 (d, J = 5.0 Hz, 2H), 2.32 (s, 4H), 1.84 (d, J = 12.3 Hz, 1H), 1.69 - 1.38 (m, 5H), 1.38 - 1.06 (m, 3H), 0.91 - 0.77 (m, 3H).

[0154] (34) Synthesis of compound ZT-1N

[0155]

[0156] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly introduced into the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then cyclobutylamine (0.46 g, 6.44 mmol) was added. After addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder of the compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 EA. The organic layers were combined. The organic layer was directly triturated and chromatographed on a basic alumina column (methylene chloride: methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ7.98 - 7.96(dd,J=4.8,1.3Hz,1H),7.93 - 7.79(dd,J=45.6,8.1Hz,2H),7.62 - 7.61(d,J=2.1Hz,1H),7.45(d,J=31.0Hz,1H),7.32 - 7.30(d,J=8.2Hz,2H),4.20 - 4.10(h,J=9.0,8.5Hz,1H),3.59 - 3.57(t,J=4.0Hz,4H),3.47 - 3.45(d,J=7.7Hz,2H),2.38 - 2.35(d,J=5.0Hz,4H),2.28 - 2.17(m,2H),2.00 - 1.87(m,2H),1.71 - 1.55(m,2H).

[0157] (35) Synthesis of compound ZT-1O

[0158]

[0159] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly introduced into the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then n-butylamine (0.47 g, 6.44 mmol) was added. After addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated and chromatographed on a basic alumina column (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ7.98(s,1H),7.93-7.79(m,2H),7.61(d,J=2.2Hz,1H),7.35-7.26(m,2H),7.14(s,1H),3.59-3.58(t,J=4.6Hz,4H),3.48-3.46(d,J=6.2Hz,2H),3.16-3.06(m,2H),2.37(s,4H),1.51-1.41(m,2H),1.39-1.29(m,2H),0.92-0.88(td,J=7.2,3.8Hz,3H).

[0160] (36) Synthesis of compound ZT-1P

[0161]

[0162] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then tert-butylamine (0.47 g, 6.44 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h, and then filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V) was carried out. 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.56 (d, J = 50.4 Hz, 1H), 8.00 (s, 1H), 7.96 - 7.79 (m, 2H), 7.60 (s, 1H), 7.29 (d, J = 7.8 Hz, 2H), 7.05 (s, 1H), 3.57 - 3.59 (t, J = 4.6 Hz, 4H), 3.45 - 3.47 (d, J = 4.6 Hz, 2H), 2.36 (s, 4H), 1.29 - 1.33 (d, J = 13.6 Hz, 9H).

[0163] (37) Synthesis of compound ZT-1Q

[0164]

[0165] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly introduced into the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then isopropylamine (0.38 g, 6.44 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h, and then filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V) was carried out. 1 H NMR(400MHz,DMSO-d 6 )δ8.73 - 8.66(d,J=27.9Hz,1H),7.99 - 7.98(d,J=5.2Hz,1H),7.81 - 7.79(d,J=8.2Hz,2H),7.61 - 7.60(d,J=2.1Hz,1H),7.32 - 7.29(d,J=8.2Hz,2H),7.13(dd,J=21.9,7.5Hz,1H),3.88 - 3.74(m,1H),3.59 - 3.57(t,J=4.6Hz,4H),3.47 - 3.45(d,J=5.7Hz,2H),2.36(s,4H),1.15 - 1.08(dd,J=21.5,6.5Hz,6H).

[0166] (38) Synthesis of compound ZT-1S

[0167]

[0168] At room temperature, compound 7a (300 mg, 0.92 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly used in the next reaction. Compound [7a] was dissolved in 15 mL of DMF solution, and then cycloheptylamine (0.73 g, 6.44 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powdered compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powdered solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated and chromatographed on a basic alumina column (dichloromethane:methanol = 120 / 1, 100 / 1, 80 / 1, 60 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ11.01-10.93(d,J=29.2Hz,1H),10.28-10.27(d,J=1.4Hz,1H),10.25-10.04(m,2H),9.89(d,J=2.1Hz,1H),9.61-9.57(m,2H),9.42(s,1H),6.00-5.92(dq,J=8.4,4.2Hz,1H),5.88-5.86(t,J=4.6Hz,4H),5.76-5.74(d,J=6.6Hz,2H),4.65(s,4H),4.20-4.05(m,2H),3.92-3.70(m,9H).

[0169] (39) Synthesis of compound ZT-2A

[0170]

[0171] At room temperature, dissolve compound 7b (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Detect that the reaction is complete by TLC. Cool the temperature of the reaction solution to room temperature, and then concentrate the solvent under reduced pressure. Without separation and purification, directly and quickly put it into the next reaction. Dissolve compound [7b] in 15 mL of DMF solution, and then add cyclohexylamine (0.64 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Detect that the reaction is complete by TLC. Cool the temperature of the reaction solution to room temperature, and then pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly make sand from the organic layer and perform column chromatography on basic alumina (methylene chloride: methanol = 100 / 1, 80 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ8.72(s,1H),8.04(s,1H),8.01 - 7.81(m,2H),7.67(s,1H),7.36 - 7.34(d,J=7.8Hz,2H),7.16(s,1H),3.58 - 3.51(m,3H),2.44(s,4H),1.91(d,J=11.0Hz,2H),1.77(d,J=11.9Hz,2H),1.57(d,J=11.1Hz,5H),1.40 - 1.18(m,7H).

[0172] (40) Synthesis of compound ZT - 2B

[0173]

[0174] At room temperature, compound 7b (300 mg, 0.93 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. Then the temperature of the reaction solution was cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7b] was dissolved in 15 mL of DMF solution, and then cyclopentylamine (0.55 g, 6.52 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. Then the temperature of the reaction solution was cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powdered compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powdered solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated and purified by column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ8.76(s,1H),7.99(d,J=2.7Hz,1H),7.83(dd,J=39.8,7.9Hz,2H),7.60(d,J=1.8Hz,1H),7.28(dd,J=8.0,6.0Hz,2H),7.14(s,1H),3.95(dq,J=12.6,6.6Hz,1H),3.42(d,J=5.4Hz,2H),2.32(d,J=6.6Hz,4H),1.88(ddt,J=31.4,12.8,5.4Hz,2H),1.68(ddq,J=9.0,6.1,3.6,3.0Hz,2H),1.50(tq,J=12.2,6.7,5.1Hz,7H),1.39(q,J=6.9,5.1Hz,2H).

[0175] (41) Synthesis of compound ZT-2C

[0176]

[0177] At room temperature, dissolve compound 7b (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and concentrate the solvent under reduced pressure. Without separation and purification, directly and quickly add it to the next reaction. Dissolve compound [7b] in 15 mL of DMF solution, and then add cyclopropylamine (0.37 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly make sand from the organic layer and perform column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V). 1H NMR (400 MHz, Methanol-d 4 ) δ 8.02 (s, 1H), 7.82 - 7.72 (s, 2H), 7.48 (s, 1H), 7.37 - 7.36 (d, J = 7.8 Hz, 2H), 3.66 (s, 2H), 2.69 (s, 1H), 2.60 (s, 4H), 1.657 - 1.64 (t, J = 5.7 Hz, 4H), 1.53 - 1.50 (d, J = 13.2 Hz, 2H), 0.87 - 0.82 (q, J = 7.1, 6.5 Hz, 2H), 0.61 - 0.57 (m, 2H).

[0178] (42) Synthesis of compound ZT-2F

[0179]

[0180] At room temperature, dissolve compound 7b (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and concentrate the solvent under reduced pressure. Without separation and purification, directly and rapidly add it to the next reaction. Dissolve compound [7b] in 15 mL of DMF solution, and then add 2-methylcyclohexylamine (0.73 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and pour the reaction solution into 150 mL of ice water. A large amount of fine powdered compound precipitates. Stir the solution for 1 h, and filter by suction, but the fine powdered solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly form sand from the organic layer, and perform column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V). 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.98 (s, 1H), 7.94 - 7.79 (m, 2H), 7.62 (s, 1H), 7.33 - 7.31 (d, J = 7.8 Hz, 2H), 6.97 (s, 1H), 3.54 (t, J = 10.5 Hz, 1H), 3.36 (s, 2H), 2.50 - 2.31 (m, 4H), 1.88 - 1.78 (m, 1H), 1.70 (d, J = 13.0 Hz, 1H), 1.64 - 1.48 (m, 5H), 1.43 (d, J = 12.0 Hz, 4H), 1.19 (d, J = 35.0 Hz, 4H), 0.96 - 0.81 (m, 4H).

[0181] (43) Synthesis of compound ZT-2J

[0182]

[0183] At room temperature, compound 7b (300 mg, 0.93 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7b] was dissolved in 15 mL of DMF solution, and then 4-methoxyaniline (0.80 g, 6.52 mmol) was added. After addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V) was carried out.

[0184] (44) Synthesis of compound ZT-2M

[0185]

[0186] At room temperature, compound 7b (300 mg, 0.93 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7b] was dissolved in 15 mL of DMF solution, and then 4-methylcyclohexylamine (0.73 g, 6.52 mmol) was added. After addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V) was carried out. 1 H NMR(400MHz,DMSO-d 6)δ 9.55 - 9.44 (d, J = 41.2 Hz, 1H), 9.13 - 8.99 (d, J = 53.4 Hz, 1H), 8.05 (s, 1H), 7.88 - 7.80 (dd, J = 26.5, 7.6 Hz, 3H), 7.62 (s, 1H), 7.43 - 7.38 (t, J = 9.3 Hz, 3H), 7.29 - 7.27 (d, J = 8.2 Hz, 3H), 6.89 - 6.87 (d, J = 9.0 Hz, 2H), 3.71 (s, 4H), 3.43 (s, 2H), 2.34 (s, 4H), 1.51 (s, 4H), 1.40 (s, 2H).

[0187] (45) Synthesis of Compound ZT - 2N

[0188]

[0189] At room temperature, dissolve compound 7b (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, and then concentrate the solvent under reduced pressure. Without separation and purification, directly and rapidly add it to the next reaction. Dissolve compound [7b] in 15 mL of DMF solution, and then add cyclobutylamine (0.48 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, and then pour the reaction solution into 150 mL of ice water. A large amount of fine powdered compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powdered solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly make sand from the organic layer and perform column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V). 1 H NMR (400 MHz, DMSO - d 6 )δ 7.99 (s, 1H), 7.98 - 7.73 (m, 2H), 7.59 (s, 1H), 7.28 (d, J = 7.6 Hz, 2H), 7.03 (s, 1H), 3.46 (s, 2H), 3.39 (s, 2H), 2.36 (s, 4H), 1.51 (t, J = 5.6 Hz, 4H), 1.40 (d, J = 5.0 Hz, 2H), 1.32 (d, J = 8.6 Hz, 9H).

[0190] (46) Synthesis of Compound ZT - 2P

[0191]

[0192] At room temperature, dissolve compound 7b (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the temperature of the reaction solution to room temperature, and then concentrate the solvent under reduced pressure. Without separation and purification, directly and rapidly put it into the next reaction. Dissolve compound [7b] in 15 mL of DMF solution, and then add tert-butylamine (0.48 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the temperature of the reaction solution to room temperature, and then pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL×3 EA. Combine the organic layers. Directly make sand from the organic layer and perform column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ7.99(s,1H),7.89 - 7.77(d,J=73.8Hz,2H),7.59(s,1H),7.28(d,J=7.7Hz,2H),7.03(s,1H),3.46(s,2H),2.36(s,4H),1.51(t,J=5.6Hz,4H),1.44 - 1.27(m,10H).

[0193] (47) Synthesis of compound ZT-2Q

[0194]

[0195] At room temperature, compound 7b (300 mg, 0.93 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7b] was dissolved in 15 mL of DMF solution, and then isopropylamine (0.38 g, 6.52 mmol) was added. After addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC until completion. Then the temperature of the reaction solution was cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated and purified by column chromatography on basic alumina (methylene chloride:methanol = 100 / 1, 80 / 1, V / V). 1 1H NMR (400 MHz, DMSO-d 6 ) δ 7.95 (s, 1H), 7.91 - 7.70 (m, 2H), 7.54 (d, J = 8.2 Hz, 1H), 7.25 (d, J = 8.1 Hz, 2H), 7.06 (t, J = 11.0 Hz, 1H), 3.78 (q, J = 6.8 Hz, 1H), 2.32 (s, 4H), 1.47 (p, J = 5.6 Hz, 4H), 1.35 (q, J = 6.0 Hz, 2H), 1.09 (t, J = 9.4 Hz, 6H).

[0196] (48) Synthesis of compound ZT-2S

[0197]

[0198] At room temperature, dissolve compound 7b (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, and then concentrate the solvent under reduced pressure. Without separation and purification, directly and quickly put it into the next reaction. Dissolve compound [7b] in 15 mL of DMF solution, and then add cycloheptylamine (0.74 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, and then pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly granulate the organic layer and perform column chromatography on basic alumina (methylene chloride: methanol = 100 / 1, 80 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ7.97(s,1H),7.88(s,2H),7.56(s,1H),7.27(d,J=7.9Hz,2H),7.12(s,1H),3.73-3.62(m,1H),3.41(s,2H),2.32(s,4H),1.86(s,2H),1.67-1.34(m,15H).

[0199] (49) Synthesis of compound ZT-4A

[0200]

[0201] At room temperature, dissolve compound 7d (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, and then concentrate the solvent under reduced pressure. Without separation and purification, directly and quickly put it into the next reaction. Dissolve compound [7d] in 15 mL of DMF solution, and then add cyclohexylamine (0.67 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the reaction solution to room temperature, and then pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly granulate the organic layer and perform column chromatography on basic alumina (methylene chloride: methanol = 80 / 1, 60 / 1, V / V).1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.72 (s, 1H), 7.96 (s, 1H), 7.86 - 7.85 (d, J = 7.8 Hz, 2H), 7.54 (s, 1H), 7.30 - 7.28 (d, J = 7.8 Hz, 2H), 7.09 (s, 1H), 3.52 (s, 2H), 3.46 (s, 1H), 2.48 - 2.42 (m, 4H), 1.85 - 1.83 (d, J = 9.8 Hz, 2H), 1.75 - 1.66 (m, 2H), 1.57 (d, J = 12.3 Hz, 1H), 1.30 - 1.16 (m, 5H), 0.98 (t, J = 7.1 Hz, 6H).

[0202] (50) Synthesis of Compound ZT-4B

[0203]

[0204] At room temperature, dissolve compound 7d (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the temperature of the reaction solution to room temperature, and then concentrate the solvent of the reaction solution under reduced pressure. Without separation and purification, directly and quickly put it into the next reaction. Dissolve compound [7d] in 15 mL of DMF solution, and then add cyclopentylamine (0.58 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Detect the completion of the reaction by TLC. Cool the temperature of the reaction solution to room temperature, and then pour the reaction solution into 150 mL of ice water. A large amount of fine powdery compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powdery solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly make sand from the organic layer and perform column chromatography on basic alumina (methylene chloride:methanol = 80 / 1, 60 / 1, V / V). 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.74 (s, 1H), 7.98 (s, 1H), 7.93 - 7.79 (dd, J = 47.8, 6.2 Hz, 2H), 7.63 (s, 1H), 7.38 - 7.36 (d, J = 7.8 Hz, 2H), 7.13 (s, 1H), 4.01 - 3.92 (m, 1H), 3.72 (s, 2H), 2.63 (d, J = 17.6 Hz, 4H), 1.96 - 1.86 (m, 2H), 1.72 - 1.59 (m, 2H), 1.57 - 1.36 (m, 4H), 1.07 - 1.03 (t, J = 6.2 Hz, 6H).

[0205] (51) Synthesis of Compound ZT-4C

[0206]

[0207] At room temperature, dissolve compound 7d (300 mg, 0.93 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and concentrate the solvent under reduced pressure. Without separation and purification, directly and quickly add it to the next reaction. Dissolve compound [7d] in 15 mL of DMF solution, and then add cyclopropylamine (0.39 g, 6.52 mmol). After addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Monitor the reaction by TLC until completion. Then cool the reaction solution to room temperature, and pour the reaction solution into 150 mL of ice water. A large amount of fine powder-like compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powder-like solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly make sand from the organic layer and perform column chromatography on basic alumina (methylene chloride:methanol = 80 / 1, 60 / 1, V / V). 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.99 (d, J = 8.2 Hz, 1H), 7.81 - 7.79 (d, J = 7.7 Hz, 2H), 7.61 (s, 1H), 7.32 - 7.30 (d, J = 7.8 Hz, 2H), 7.17 (s, 1H), 3.61 (s, 1H), 3.33 (s, 2H), 2.65 (m, 4H), 1.36 - 0.95 (m, 6H), 0.75 - 0.71 (t, J = 7.0 Hz, 2H), 0.52 - 0.48 (p, J = 4.9, 4.5 Hz, 2H).

[0208] (52) Synthesis of Compound ZT-4F

[0209]

[0210] At room temperature, compound 7d (300 mg, 0.93 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly introduced into the next reaction. Compound [7d] was dissolved in 15 mL of DMF solution, and then 2-methylcyclohexylamine (0.77 g, 6.52 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powdery compound precipitated out. The solution was stirred for 1 h, and then filtered by suction, but the fine powdery solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 80 / 1, 60 / 1, V / V) was carried out. 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.97 (s, 1H), 7.94 - 7.73 (m, 2H), 7.61 (s, 1H), 7.34 - 7.32 (s, 2H), 6.97 (s, 1H), 3.66 - 3.59 (d, J = 14.6 Hz, 2H), 3.14 (s, 1H), 2.57 - 2.51 (m, 4H), 1.86 - 1.84 (d, J = 11.5 Hz, 1H), 1.75 - 1.72 (d, J = 14.0 Hz, 1H), 1.63 - 1.60 (d, J = 11.2 Hz, 1H), 1.51 - 1.11 (m, 6H), 1.02 (t, J = 7.0 Hz, 6H), 0.92 (d, J = 6.5 Hz, 3H).

[0211] (53) Synthesis of compound ZT-5A

[0212]

[0213] At room temperature, compound 7e (300 mg, 0.89 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly used in the next reaction. Compound [7e] was dissolved in 15 mL of DMF solution, and then cyclohexylamine (0.62 g, 6.23 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 80 / 1, 60 / 1, V / V) was carried out. 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.93 (s, 1H), 7.90 - 7.70 (m, 2H), 7.55 (s, 1H), 7.25 - 7.23 (s, 2H), 7.04 (s, 1H), 3.41 (s, 3H), 2.51 - 2.49 (p, J = 1.9 Hz, 2H), 2.45 - 2.42 (s, 2H), 2.34 - 2.25 (d, J = 19.0 Hz, 4H), 2.11 (s, 3H), 1.87 - 1.81 (d, J = 15.0 Hz, 2H), 1.67 - 1.65 (d, J = 8.0 Hz, 2H), 1.59 - 1.46 (m, 1H), 1.28 - 1.08 (m, 5H).

[0214] (54) Synthesis of compound ZT-5B

[0215]

[0216] At room temperature, compound 7e (300 mg, 0.89 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7e] was dissolved in 15 mL of DMF solution, and then cyclopentylamine (0.53 g, 6.23 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h, and then filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 80 / 1, 60 / 1, V / V) was carried out. 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.98 (s, 1H), 7.93 - 7.75 (m, 2H), 7.59 (s, 1H), 7.28 - 7.26 (d, J = 8.0 Hz, 2H), 7.10 (s, 1H), 3.99 - 3.94 (q, J = 7.0 Hz, 1H), 3.45 (s, 2H), 2.55 - 2.51 (p, J = 1.8 Hz, 2H), 2.49 - 2.45 (p, J = 1.8 Hz, 2H), 2.39 - 2.28 (d, J = 4.4 Hz, 4H), 2.14 (s, 3H), 1.94 - 1.86 (s, 2H), 1.67 (s, 2H), 1.57 - 1.40 (m, 4H).

[0217] (55) Synthesis of compound ZT-5M

[0218]

[0219] At room temperature, compound 7e (300 mg, 0.89 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7e] was dissolved in 15 mL of DMF solution, and then 4-methylcyclohexylamine (0.70 g, 6.23 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powdered compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powdered solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated and chromatographed on a basic alumina column (dichloromethane:methanol = 80 / 1, 60 / 1, V / V). 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.97 (s, 1H), 7.94 - 7.70 (m, 2H), 7.59 (s, 1H), 7.28 (d, J = 7.8 Hz, 2H), 7.06 (s, 1H), 3.45 (s, 3H), 2.55 - 2.52 (dq, J = 4.0, 2.0 Hz, 2H), 2.49 - 2.45 (m, 2H), 2.41 - 2.28 (m, 4H), 2.14 (s, 3H), 1.91 - 1.83 (t, J = 15.0 Hz, 1H), 1.74 - 1.42 (m, 4H), 1.33 - 1.20 (m, 4H), 0.90 - 0.86 (dd, J = 22.2, 6.4 Hz, 3H).

[0220] (56) Synthesis of compound ZT-6A

[0221]

[0222] At room temperature, compound 7f (300 mg, 0.89 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly introduced into the next reaction. Compound [7f] was dissolved in 15 mL of DMF solution, and then cyclohexylamine (0.62 g, 6.23 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powdered compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powdered solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, V / V) was carried out. 1 H NMR(400MHz,DMSO-d 6 )δ7.97(s,1H),7.92-7.77(m,2H),7.59(s,1H),7.29-7.27(d,J=7.8Hz,2H),7.07-7.01(d,J=24.6Hz,1H),3.44(s,1H),2.76-2.71(t,J=10.0Hz,2H),2.55-2.52(dp,J=4.0,1.8Hz,2H),2.49(s,2H),1.85(d,J=11.0Hz,3H),1.73-1.55(m,6H),1.36-1.06(m,6H),0.81(d,J=5.6Hz,3H).

[0223] (57) Synthesis of compound ZT-6B

[0224]

[0225] At room temperature, compound 7f (300 mg, 0.89 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly introduced into the next reaction. Compound [7f] was dissolved in 15 mL of DMF solution, and then cyclopentylamine (0.53 g, 6.23 mmol) was added. After addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, V / V) was carried out. 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.76 (s, 1H), 7.99 (s, 1H), 7.95 - 7.70 (m, 2H), 7.58 (s, 1H), 7.28 - 7.26 (d, J = 7.8 Hz, 2H), 7.12 (s, 1H), 4.03 - 3.91 (m, 1H), 3.50 - 3.42 (d, J = 3.9 Hz, 4H), 2.74 - 2.69 (t, J = 10.3 Hz, 2H), 1.93 - 1.82 (q, J = 13.2, 10.6 Hz, 3H), 1.72 - 1.36 (m, 10H), 0.82 - 0.79 (d, J = 5.4 Hz, 3H).

[0226] (58) Synthesis of compound ZT-6C

[0227]

[0228] At room temperature, compound 7f (300 mg, 0.89 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. TLC detected that the reaction was complete. The temperature of the reaction solution was lowered to room temperature, and then the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7f] was dissolved in 15 mL of DMF solution, and then cyclopropylamine (0.36 g, 6.23 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. TLC detected that the reaction was complete. The temperature of the reaction solution was lowered to room temperature, and then the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, V / V). 1H NMR (400 MHz, DMSO-d 6 ) δ 8.00 (s, 1H), 7.79 - 7.77 (d, J = 7.8 Hz, 2H), 7.59 (s, 1H), 7.26 - 7.24 (d, J = 7.8 Hz, 2H), 7.16 (s, 1H), 3.42 (s, 1H), 2.80 - 2.59 (m, 3H), 2.07 - 1.93 (m, 1H), 1.88 - 1.83 (t, J = 11.0 Hz, 1H), 1.66 - 1.45 (m, 5H), 0.80 (d, J = 5.3 Hz, 3H), 0.73 (d, J = 7.6 Hz, 2H), 0.52 - 0.37 (m, 2H).

[0229] (59) Synthesis of compound ZT-6F

[0230]

[0231] At room temperature, compound 7f (300 mg, 0.89 mmol) was dissolved in 20 mL of THF solution, and then CDI (298 mg, 28.5 mmol) was added. After the addition, the temperature of the reaction solution was raised to 72 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the solvent was concentrated under reduced pressure. Without separation and purification, it was directly and rapidly put into the next reaction. Compound [7f] was dissolved in 15 mL of DMF solution, and then 2-methylcyclohexylamine (0.71 g, 6.23 mmol) was added. After the addition, the temperature of the reaction solution was raised to 100 °C and heated under reflux for 16 h. The reaction was monitored by TLC and found to be complete. The temperature of the reaction solution was then cooled to room temperature, and the reaction solution was poured into 150 mL of ice water. A large amount of fine powder-like compound precipitated out. The solution was stirred for 1 h and filtered by suction, but the fine powder-like solid could not be filtered out. It was extracted with 50 mL of EA, and the aqueous layer was extracted with 25 mL × 3 of EA. The organic layers were combined. The organic layer was directly triturated, and column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, V / V) was carried out. 1 H NMR(400MHz,DMSO-d 6 )δ7.94(s,1H),7.88-7.74(dd,J=50.4,7.26-7.16Hz,2H),7.53(s,1H),7.26(s,2H),6.98(d,J=48.3Hz,1H),3.42(s,2H),3.17-3.04(m,1H),2.70(s,2H),2.52-2.47(m,2H),2.45(s,2H),1.86-1.76(m,1H),1.74-1.33(m,9H),1.19(s,3H),0.87(t,J=7.5Hz,3H),0.77(d,J=5.4Hz,3H).

[0232] (60) Synthesis of compound ZT-6M

[0233]

[0234] At room temperature, dissolve compound 7f (300 mg, 0.89 mmol) in 20 mL of THF solution, and then add CDI (298 mg, 28.5 mmol). After the addition, raise the temperature of the reaction solution to 72 °C and reflux for 16 h. Monitor the reaction completion by TLC. After the reaction solution cools down to room temperature, concentrate the solvent under reduced pressure. Without separation and purification, directly and rapidly add it to the next reaction. Dissolve compound [7f] in 15 mL of DMF solution, and then add 4-methylcyclohexylamine (0.71 g, 6.23 mmol). After the addition, raise the temperature of the reaction solution to 100 °C and reflux for 16 h. Monitor the reaction completion by TLC. After the reaction solution cools down to room temperature, pour the reaction solution into 150 mL of ice water. A large amount of fine powdered compound precipitates. Stir the solution for 1 h and filter by suction, but the fine powdered solid cannot be filtered out. Extract with 50 mL of EA, and extract the aqueous layer with 25 mL × 3 EA. Combine the organic layers. Directly granulate the organic layer and perform column chromatography on basic alumina (methylene chloride:methanol = 120 / 1, 100 / 1, 80 / 1, V / V). 1 H NMR(400MHz,DMSO-d 6 )δ8.70(s,1H),7.98-7.96(d,J=6.8Hz,1H),7.92-7.988(s,2H),7.77(s,1H),7.59(s,1H),7.29-7.27(d,J=8.2Hz,2H),7.05(s,1H),3.75-3.72(d,J=14.1Hz,2H),2.76-2.71(d,J=13.2Hz,4H),1.89-1.84(d,J=12.1Hz,3H),1.66(s,2H),1.58 -1.56(m,5H),1.48(s,2H),1.36(d,J=8.0Hz,2H),0.81-0.80(d,J=5.2Hz,6H).

[0235] II. Experimental methods

[0236] 1. Method

[0237] 1.1 Study on the inhibition of Aurora A kinase and JAK2 activities by compounds

[0238] The HTRF-KinEASE-STK kit is used for the kinase activity inhibition assay experiment. In a 384-well plate, add kinase, substrate, ATP, and buffer to the negative control group; add only substrate, ATP, and buffer to the blank control group; add kinase, substrate, ATP, and different concentrations of the test compounds to each test compound group. After reacting at room temperature for 1 h, terminate with EDTA, and add Eu3 +The STK antibody and streptavidin-XL665 were reacted for 1 hour. The fluorescence intensities at 665 nm and 615 nm were detected with a Synergy 4 multimode microplate reader using an excitation light of 340 nm, and the fluorescence ratio of 615 nm / 665 nm, namely the HTRF value, was calculated. The half maximal inhibitory concentration (IC 50 ) was calculated by fitting with GraphPad Prism 5.0 software. The reaction conditions for each kinase were as follows:

[0239] Aurora A: Aurora A, 3 ng / μL; STK substrate 2-biotin, 1 μM; ATP, 15 μM;

[0240] The substrate of CHK1 was STK substrate 1-biotin, the substrate of PKA was STK substrate 2-biotin, the substrate of AKT1 was STK substrate 3-biotin, and the substrates of the remaining kinases were all TK.

[0241] 1.2 Study on the inhibitory activity of the compound against human breast cancer cell MDA-MB-231

[0242] In this experiment, the CCK-8 cell proliferation assay was mainly used to determine the effect of the compound on the in vitro proliferation of human triple-negative breast cancer cells. Aurora A and JAK2 kinases are highly expressed in MDA-MB-231 cells. Therefore, the cells MDA-MB-231 were selected for the experiment on the effect of the compound on the in vitro proliferation of human triple-negative breast cancer cells. The experimental method was as follows:

[0243] (1) The cells were digested and counted, and a cell suspension of 4×104 cells / mL was prepared. 100 μL of the cell suspension was added to each well of a 96-well cell culture plate;

[0244] (2) The 96-well cell culture plate was placed in an incubator at 37 °C, 5%, CO 2 and cultured for 24 hours;

[0245] (3) The drug was diluted with the medium to the required working solution concentration, and 100 μL of the corresponding drug-containing medium was added to each well. At the same time, a negative control group was set up;

[0246] (4) The 96-well cell culture plate was placed in an incubator at 37 °C, 5%, CO 2 and cultured for 72 hours;

[0247] (5) The 96-well plate was stained with CCK-8, λ = 450 nm, and the OD value was measured;

[0248] 1) Add 10 μL of CCK-8 to each well and continue culturing in the incubator for 2 - 3 hours;

[0249] 2) Gently mix on a shaker for 10 minutes to remove air bubbles in the 96-well plate;

[0250] 3) At λ = 450 nm, read the OD value of each well with an enzyme-linked immunosorbent assay (ELISA) reader and calculate the inhibition rate.

[0251] (6) Calculate the inhibition rate of each group.

[0252]

[0253] According to the inhibition rate, calculate the half-maximal inhibitory concentration (IC50) using the LOGIT method. 50 The experiment was repeated more than 3 times, and the data were expressed as Mean ± SD. 1.3 Detection of intracellular Aurora A and STAT3 expression by Western blot

[0254] The human breast cancer cell line MDA-MB-231 is an adherent cell, and all were cultured in DMEM medium containing 10% fetal bovine serum (FBS). The conventional culture was placed in an incubator at 37°C and 5% CO2, and divided into a drug treatment group and a control group. In the drug treatment group, the compound concentrations were 0 μM, 10 μM, and 20 μM. After incubation for 48 h, the cells were collected and lysed. The protein concentration was determined by the BCA method, and 50 μg of protein was electrophoresed using 10% SDS-PAGE. The separated proteins were transferred to a polyvinylidene difluoride (PVDF) membrane. After blocking with 5% milk, the blot was probed with antibodies and then incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody, and detected using 2 SuperSignal West Pico Chemiluminescent substrate

[0255]

[0256] 1.4 Flow cytometry analysis of the compound-induced cell cycle arrest in human breast cancer cell line MDA-MB-231

[0256] Count the MDA-MB-231 cells. According to the counting results, take 5×10 5 cells per well and evenly spread the cell suspension in a 24-well plate, and add complete medium with a total volume of 1 mL. Set the compound concentration gradients to 0, 10, 20 μM (0, 5, 10 μM) and according to C 1 V 1 = C 2 V 2Calculate the required volume of the drug and add it to the wells seeded with cells. Set up three replicates for each concentration to ensure the reliability of the experiment. Place the well plate in the incubator and culture for 24 h. After 24 h, collect the cells into a flow cytometry tube and centrifuge at 3000 rpm for 3 min to precipitate the cells. Discard the supernatant, and while vortexing, add 70% ethanol dropwise to fully fix the MDA-MB-231 cells. Place them at 4 °C overnight. After overnight fixation, centrifuge the cells at 3000 rpm for 3 min to precipitate the cells. Discard the PBS and wash the cells twice. Add PI staining solution at 200 μL / L to the tube wall, resuspend and mix well, and incubate in the dark for half an hour. Filter the cells through a filter before loading onto the machine. Collect the cells at a low speed throughout the process. Repeat the experiment three times. Use ModFit 5.0 to process the data and record the proportion of cells in each stage of the cell cycle.

[0257] 1.5 Detection of apoptosis of compound on human breast cancer cell line MDA-MB-231 by flow cytometry

[0258] Cell plating and drug treatment are the same as in Section 1.2.4 of this part, but set the compound concentration gradients as 0, 5, and 10 μM. Place the 24-well plate in the incubator and culture for 48 h. After 48 h, collect the cells into a flow cytometry tube and centrifuge at 3000 rpm for 3 min to precipitate the cells. Discard the supernatant. Add 200 μL of 1×Binding Buffer to each tube to resuspend the cells, and add 1 μL of APC-AnnexinV and 2 μL of 7-AAD to the cell suspension and incubate in the dark at room temperature for 15 min. After incubation, place the flow cytometry tube on the flow cytometer for detection. Repeat the experiment three times. Use FlowJo 7.6 to analyze the data and record the proportion of apoptotic cells at each concentration.

[0259] 1.6 Data processing

[0260] All data are expressed as mean ± standard deviation (Mean ± SD), and statistical analysis is performed using SPSS19.0 software. The comparison between two groups uses an unpaired two-tailed t-test; the comparison among multiple groups uses a one-way analysis of variance (one-way ANOVA) with a completely randomized design. The significance level of the hypothesis test is determined according to α = 0.05. P < 0.05 indicates that the difference is statistically significant.

[0261] 2. Results and discussion

[0262] 2.1 Study on the inhibition of Aurora A / JAK2 kinase activity by the compound

[0263] Use the HTRF-KinEASE-STK luminescent kinase assay kit to determine the activity of the compound against Aurora A and JAK2 kinases. The results are shown in Table 1.

[0264] Table 1 Inhibitory activities of compounds against Aurora A and JAK2 kinases

[0265]

[0266]

[0267] a. Positive control drugs: MZ-1C and MD-1 (derived from CN111004220A); b. Not Test

[0268] As can be seen from Table 1, the tested target compounds all have good inhibitory effects on Aurora A kinase and JAK2 kinase. Among them, compounds ZT-1B, ZT-2F, ZT-2N, ZT-2S, ZT-2P, ZT-2Q, ZT-4A, ZT-4B, ZT-4C, ZT-4F, ZT-6F and ZT-6M have significant inhibitory activities. The IC 50 of ZT-2F against Aurora A and JAK2 kinases are 0.005 ± 0.0004 μM and 0.021 ± 0.002 μM respectively.

[0269] 2.2 Determination of the inhibitory rate of compounds at a single concentration by CCK-8 method

[0270] In the experiment to detect the effect of compounds on the in vitro proliferation of human triple-negative breast cancer cells, human triple-negative breast cancer cells MDA-MB-231 were selected for activity detection. The cancer cells in the logarithmic growth phase were seeded in a 96-well cell culture plate, and 100 μL of cell suspension was added to each well. After culturing for 24 h, the drugs were administered at a concentration of 10 μM. After the drugs acted for 72 h, CCK-8 staining was performed to measure the OD value and calculate the inhibitory rate. The results are shown in Table 2.

[0271] Table 2 Growth inhibitory rates of compounds against MDA-MB-231 (10 μM)

[0272]

[0273]

[0274] a. Positive control drugs: MZ-1C, paclitaxel, Ruxolitinib and MD-1 (derived from CN111004220A); b. NotTest

[0275] As can be seen from Table 2, most of the ZT-1 / 2 series compounds showed good inhibitory activity against MDA-MB-231 cells under the condition of 10 μM, higher than that of the positive control drugs MZ-1C, Ruxolitinib and paclitaxel. The inhibition rates of ZT-2A, ZT-2B, ZT-2F, ZT-2M, ZT-2A and ZT-2B reached more than 90%, among which the inhibition rates of ZT-2F and ZT-2M were as high as 95.22% and 96.22% respectively.

[0276] 2.3 Determination of the IC of compounds by CCK-8 method 50

[0277] According to the above results, compounds with a single-concentration inhibition rate against human triple-negative breast cancer cells MDA-MB-231 exceeding 50% were selected for the determination of IC 50 values. Three replicate wells were set for each drug concentration, and the test results are shown in Table 3.

[0278] Table 3 IC of ZT series compounds against cells MDA-MB-231 50

[0279]

[0280]

[0281] a. Positive control drugs: MZ-1C, paclitaxel and Ruxolitinib; b.Not Test

[0282] As can be seen from Table 3: The selected compounds all had good anti-cell proliferation activity against MDA-MB-231 cells, with IC 50 values between 5 and 10 μM, and better than the positive control drug MZ-1C. Among them, the IC 50 values of compounds ZT-1B, ZT-2F and ZT-2M were 6.565 μM, 5.247 μM and 5.860 μM respectively, which were worthy of further study.

[0283] 2.4 Detection of the expression of Aurora A / STAT3 protein by Western Blot

[0284] Based on the comprehensive results of anti-triple-negative breast cancer cell proliferation and kinase inhibitory activity, compounds ZT-1B, ZT-2F and ZT-2M were selected for further experimental research. Western Blot was used to detect the inhibition of the expression of intracellular Aurora A, JAK2 and STAT3 by compounds ZT-1B, ZT-2F and ZT-2M, and the results are as Figure 1 shown.

[0285] From Figure 1It can be seen that at a concentration of 10 μM, compound ZT-1B significantly reduces the expression levels of Aurora A and STAT3 proteins; compound ZT-2M reduces the expression level of STAT3 protein at a concentration of 10 μM; compound ZT-2F can significantly reduce the protein levels of Aurora A, JAK2, and STAT3 proteins and inhibit the phosphorylation of p-JAK2 and p-STAT3 at a concentration of 10 μM.

[0286] 2.5 Cell cycle arrest experiment

[0287] Inhibiting the overexpression of Aurora A kinase and JAK2 can induce G2 phase arrest and thus apoptosis in MDA-MB-231 cells. Therefore, in this invention, flow cytometry was used to detect the effects of compounds ZT-1B, ZT-2F, and ZT-2M on the cell cycle progression of MDA-MB-231 cells. Among them, the results of ZT-2F on the cell cycle progression of MDA-MB-231 cells are as Figure 2 shown.

[0288] Compound ZT-1B induces 51.14% of the cells to arrest in the G2 phase at a concentration of 10 μM and 64.67% of the cells to arrest in the G2 phase at a concentration of 20 μM; compound ZT-2F induces 15.06% of the cells to arrest in the G2 phase at a concentration of 5 μM and 17.75% of the cells to arrest in the G2 phase at a concentration of 10 μM, as Figure 2 ; compound ZT-2M induces 26.70% of the cells to arrest in the G2 phase at a concentration of 5 μM and 53.81% of the cells to arrest in the G2 phase at a concentration of 10 μM, and induces dose-dependent G2 phase arrest of MDA-MB-231 cells.

[0289] 2.6 Apoptosis experiment

[0290] Inhibiting the overexpression of Aurora A kinase and JAK2 can induce apoptosis in MDA-MB-231 cells. To study the effect of the compound on the apoptosis of MDA-MB-231 cells, flow cytometry was used to detect the number of cells double-stained with Annexin V-APC-A and 7-AAD-A. Early apoptotic cells were defined as Annexin 7-AAD-A, late apoptotic cells were defined as Annexin APC-A, and the total percentage of apoptotic cells was the sum of early apoptotic cells and late apoptotic cells.

[0291] Compared with the control group, the early apoptosis rate of compound ZT-1B in MDA-MB-231 cells treated at a concentration of 5 μM was 44.4%, and the late apoptosis rate was 16.9%. At a concentration of 10 μM, the early apoptosis rate was 35.3% and the late apoptosis rate was 21.0%. For compound ZT-2F at a concentration of 5 μM, the early apoptosis rate of MDA-MB-231 cells treated was 58.8%, and the late apoptosis rate was 5.08%. At a concentration of 10 μM, the early apoptosis rate was 48.4% and the late apoptosis rate was 6.78%, as Figure 3 shown; for compound ZT-2M at a concentration of 5 μM, the early apoptosis rate of MDA-MB-231 cells treated was 78.4%, and the late apoptosis rate was 3.14%. At a concentration of 10 μM, the early apoptosis rate was 73.9% and the late apoptosis rate was 2.83%. Among the three MDA-MB-231 cell lines, the percentage of apoptotic cells increased with the increase of drug concentration after 48 h of treatment.

[0292] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: it is still possible to modify the technical solutions described in the foregoing embodiments, or to equivalently replace some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof, in, R 1 represents X, heteroalicyclic group or substituted heteroalicyclic group, wherein the substituted heteroalicyclic group may be arbitrarily substituted or polysubstituted by the following substituents: C1-C4 alkyl, C1-C4 alkoxy, hydroxyl or halogen; X represents the following groups: R 3 represents C1-C4 alkyl, C1-C4 haloalkyl or phenyl; R 4 represents C1-C4 alkyl, C1-C4 haloalkyl or phenyl; R 2 represents C3-C10 cycloalkyl, substituted C3-C10 cycloalkyl, C1-C6 alkyl, phenyl or substituted phenyl; the substituted C3-C10 cycloalkyl or substituted phenyl may be arbitrarily mono- or poly-substituted by the following substituents: C1-C4 alkyl, C1-C4 alkoxy, hydroxyl or halogen.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 represents X, a morpholine ring, a piperidine ring, a piperazine ring, a tetrahydropyrrole ring, a substituted morpholine ring, a substituted piperidine ring or a substituted piperazine ring, wherein the substituted morpholine ring, the substituted piperidine ring or the substituted piperazine ring may be arbitrarily substituted or polysubstituted by the following substituents: methyl, ethyl, methoxy or ethoxy; R 3 Represents methyl or ethyl; R 4 Represents methyl or ethyl.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein R 1 represents X, a morpholine ring, a piperidine ring, a piperazine ring, a substituted piperidine ring or a substituted piperazine ring, wherein the substituted piperidine ring or the substituted piperazine ring may be arbitrarily substituted or polysubstituted by the following substituents: methyl or ethyl; preferably, R 1 Represents the following groups:

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 2 represents cyclopropane, cyclobutane, cyclopentane, substituted cyclopentane, cyclohexane, substituted cyclopentane, cycloheptane, propyl, isopropyl, tert-butyl, phenyl, substituted phenyl; the substituted cyclopentane, substituted cyclopentane or substituted phenyl may be arbitrarily mono- or poly-substituted by the following substituents: methyl, ethyl, methoxy, ethoxy, fluorine or chlorine.

5. The compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein R 2 represents cyclopropane, cyclobutane, cyclopentane, cyclohexane, substituted cyclopentane, cycloheptane, propyl, isopropyl, tert-butyl, phenyl, substituted phenyl; the substituted cyclopentane, substituted cyclopentane or substituted phenyl may be arbitrarily mono- or poly-substituted by the following substituents: methyl, methoxy or fluorine.

6. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein R 2 Represents the following groups:

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 Represents the following groups: R 2 Represents the following groups:

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein The compound is selected from:

9. A pharmaceutical composition, comprising the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof as an active ingredient or a main active ingredient, and supplemented with a pharmaceutically acceptable carrier.

10. Use of the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 8 in the preparation of drugs for tumor-related diseases, in particular in the preparation of dual-target inhibitors of Aurora A kinase and JAK2 kinase.

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