CN113582906A - Difluorophenyl ring propylamine compound and preparation method and application thereof - Google Patents

Abstract

The invention provides a difluorophenyl cyclopropylamine compound which is characterized by being a 3, 4-difluorophenyl cyclopropylamine derivative or a pharmaceutically acceptable salt thereof, wherein the general formula of the 3, 4-difluorophenyl cyclopropylamine derivative is

Description

Difluorophenyl ring propylamine compound and preparation method and application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a difluorophenyl alanine compound and a preparation method thereof, which are applied to the development of an anti-tumor drug taking histone lysine specific demethylase 1(LSD1) as a target.

Background

Lysine-specific demethylase 1(LSD1) was the first histone demethylase to be found, which specifically demethylates and demethylates the methyl groups at the H3K4 and H3K9 sites by a Flavin Adenine Dinucleotide (FAD) -dependent mechanism, thereby regulating gene expression and transcriptional activity. Current studies demonstrate that LSD1 is overexpressed in a variety of tumor cells. LSD1 can regulate gene expression by activating or inhibiting chromatin domains through histone demethylation, and regulate tumor development and development by influencing the expression of factors essential for cell proliferation and differentiation. Therefore, LSD1 is gaining increasing attention as an attractive target for anticancer drugs.

Since LSD1 belongs to the family of amine oxidases and has structural and sequence similarities with monoamine oxidases (MAOs) a and B, in earlier studies, MAOs inhibitors were used to screen for backbone structures with LSD1 inhibitory activity, with the best phenylpropylamine (TCP) activity. Among the LSD1 inhibitors that have been reported so far, there are 6 LSD1 inhibitors with TCP structure: TCP, ORY-1001, GSK-2879552, INCB059872, IMG-7289 and ORY-2001, have been used in clinical trial studies. Therefore, TCP is more and more concerned as a pharmacophore of an LSD1 inhibitor, and has certain guiding significance for developing a novel LSD1 small molecule inhibitor based on the structure expansion and the synthesis of diverse small molecule compounds of TCP.

Disclosure of Invention

In view of the above, in order to develop and utilize the existing clinical drug resources, the present invention needs to provide a novel difluorophenyl cyclopropylamine compound, and a preparation method and an application thereof.

Therefore, the invention aims to provide difluorophenyl cyclopropylamine compounds and a preparation method thereof, so as to open up a new way for searching a new antitumor drug based on an LSD1 target.

The invention also aims to provide a 3, 4-difluorophencyclylamine derivative or a pharmaceutically acceptable salt thereof for preparing the LSD1 inhibitor.

In order to achieve the above object, the present invention provides a difluorophenylalanyl amine compound, which is a 3, 4-difluorophenylalanyl amine derivative or a pharmaceutically acceptable salt thereof, wherein the general formula of the 3, 4-difluorophenylalanyl amine derivative is as follows:

wherein the group R in the formula¹Is H, contains C_1～6Alkyl group of (C)_3～7Cycloalkyl, phenyl or substituted phenyl of (a);

radical R²Is H, contains C_1～6Alkyl group of (C)_1～6Halogenoalkyl of (C)_3～7Cycloalkyl of, containing C_3～6Halocycloalkyl, phenyl or substituted phenyl of (a);

radical R³Is H, contains C_1～6Alkyl, phenyl or substituted phenyl, halogen, containing C_1～6Alkoxy, cyano, nitro, trifluoromethyl;

radical R⁴Is hydroxy, containing C_1～6Alkoxy, amino or substituted amino, anilino, benzylamino or substituted benzylamino, piperidinyl, piperazinyl, N-substituted piperazinyl, N-disubstituted amino, C_3～6A cyclic amino group.

Wherein "substituted phenyl" herein means containing C_1～6Alkyl-substituted phenyl, halophenyl, cyano-substituted phenyl, nitro-substituted phenyl or amino-substituted phenyl. As used herein, "substituted amino" refers to a group containing C_1～6Alkylamino or halogen-substituted amino of (a). As used herein, the term "substituted benzylamino" refers to benzylamino wherein the phenyl group is substituted at the carbon atom, specifically to benzylamino substituted with halogen, C-containing groups_1～3Alkyl-substituted benzylamino group of (A), C_1～3Alkoxy-substituted benzylamino, cyano-substituted benzylamino or nitro-substitutedA benzylamino group.

In the above formula VI and formula VII, the above formula VII corresponds to the hydrochloride of formula VI. Namely, the 3, 4-difluorophenyl cyclopropylamine derivative is a compound represented by the general formula VI or a hydrochloride thereof.

Preferably, the group R¹Is H, methyl, ethyl, propyl, cyclopropyl or phenyl.

Preferably, the group R²Is H, methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, phenyl, or halophenyl. Wherein the halophenyl is 4-fluorophenyl, 4-chlorophenyl or 4-bromophenyl.

Preferably, the group R³Is H, 4-methyl, 4-ethyl, 4-propyl, 4-butyl, 4-fluoro, 4-bromo, 3-phenyl, 3-benzyl, 3-methoxy, 3-ethoxy, 3-cyano, 3-nitro, or 4-trifluoromethyl; wherein "4-" means a group R³In the 4-position of the phenyl radical, "3-" is a radical R³At the 3-position of the phenyl group.

The group R⁴Is hydroxyl, methoxy, ethoxy, pentoxy, amino, fluoro-amino, methylamino, ethylamino,

(N-tert-butoxycarbonylpiperazinyl),

Based on the above, the 3, 4-difluorophencyclylamine derivative is one of the compounds VI-1 to VI-20 having the general formula VI and the following groups:

VI-1:R¹＝H,R²＝H,R³＝H,

VI-2:R¹＝H,R²＝H,R³＝H,

VI-3:R¹＝H,R²＝H,R³＝H,

VI-4:R¹＝H,R²＝H,R³＝H,

VI-5:R¹＝H,R²＝H,R³＝H,

VI-6:R¹＝H,R²＝H,R³＝H,

VI-7:R¹＝H,R²＝H,R³＝H,

VI-8:R¹＝H,R²＝H,R³＝H,

VI-9:R¹＝H,R²＝H,R³＝H,

VI-10:R¹＝H,R²＝H,R³＝H,

VI-11:R¹＝H,R²＝H,R³＝H,

VI-12:R¹＝H,R²＝H,R³＝H,

VI-13:R¹＝H,R²＝H,R³＝H,

VI-14:R¹＝H,R²＝H,R³＝H,

VI-15:R¹＝H,R²＝H,R³＝H,

VI-16:R¹＝H,R²＝H,R³＝H,

VI-17:R¹＝H,R²＝H,R³＝H,

VI-18:R¹＝H,R²＝H,R³＝H,

VI-19:R¹＝H,R²＝H,R³＝H,

VI-20:R¹＝H,R²＝H,R³＝H,

based on the above, the 3, 4-difluorophencyclylamine derivative is one of the compounds VI-1 to VI-13 having the general formula VII and the following groups:

VII-1:R¹＝H,R²＝H,R³＝H,

VII-2:R¹＝H,R²＝H,R³＝H,

VII-3:R¹＝H,R²＝H,R³＝H,

VII-4:R¹＝H,R²＝H,R³＝H,

VII-5:R¹＝CH₃,R²＝H,R³＝H,

VII-6:

R²＝H,R³＝H,

VII-7:

R²＝H,R³＝H,

VII-8:R¹＝H,R²＝CH₃,R³＝H,

VII-9:R¹＝H,

R³＝H,

VII-10:R¹＝H,

R³＝H,

VII-11:R¹＝H,R²＝H,R³＝4-CH₃,

VII-12:R¹＝H,R²＝H,R³＝4-F,

VII-13:R¹＝H,R²＝H,R³＝4-Br,

the invention also provides a preparation method of the difluorophenyl ring propylamine compound, which comprises the following steps:

the preparation of the intermediate IV is that 3, 4-difluorophenyl cyclopropylamine and ethyl glyoxylate are dissolved in a toluene mixed solvent and then react with an indole compound III at normal temperature to prepare the intermediate IV; wherein the toluene mixed solvent mainly comprises toluene and hexafluoroisopropanol (abbreviated as HFIP), and the structural general formula of the indole compound III is as follows:

the structural general formula of the intermediate IV is as follows:

preparation of intermediate V the intermediate IV is first reacted with aqueous lithium hydroxide solution in an organic solvent at room temperature; removing the organic solvent, and adjusting the pH value to 3-4 by using hydrochloric acid to precipitate a solid; then carrying out suction filtration and separation to obtain an intermediate V; wherein the organic solvent is tetrahydrofuran or dioxane, and the structural general formula of the intermediate V is

Preparation of general formula VI the intermediates V and HATU (Chinese name: 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate) were dissolved in DMF (Chinese name: N, N-dimethylformamide) at ambient temperature to form a system S1; reacting an amine compound R⁴H and organic base DIPEA (Chinese name: N, N-diisopropylethylamine) are dissolved in DMF to form a system S2; the system S2 was added dropwise to the system S1 and reacted at room temperature to produce the product of general formula VI.

In the preparation step of the intermediate IV, HFIP in the toluene mixed solvent is mainly used for promoting the condensation reaction of 3, 4-difluorophenylalanyl amine and ethyl glyoxylate to generate imine and the subsequent addition reaction of the imine and an indole compound III; toluene is primarily used to increase the solubility of the reaction substrate. The synthesis mechanism of the intermediate IV is as follows: the intermediate IV is formed by pre-activating 3, 4-difluorophenylalanyl amine and ethyl glyoxylate in a toluene mixed solvent consisting of HFIP and toluene to form an imine intermediate, adding indole compound III, and directly combining three molecules into one molecule through an addition reaction promoted by HFIP.

Based on the preparation method, the preparation of the intermediate IV comprises the following steps: uniformly stirring 3, 4-difluorophenylalanyl amine and ethyl glyoxylate with the mixed solvent at normal temperature according to the molar ratio of 1: 1-1: 3, wherein the volume ratio of toluene to hexafluoroisopropanol in the mixed solvent is 1: 1-5: 1; and then adding the substituted indole III, continuing stirring at normal temperature for 6-12 h, monitoring by TLC (thin layer chromatography) to complete reaction, cooling to room temperature, adding silica gel, stir-frying, and performing column chromatography separation to obtain the intermediate IV.

Based on the preparation method, the preparation method of the intermediate V comprises the following steps: dissolving the intermediate IV in dioxane, and stirring in an ice bath at 0 ℃ to form an intermediate IV mixed solution; then dissolving lithium hydroxide in water to form the lithium hydroxide aqueous solution, wherein the molar ratio of the intermediate IV to the lithium hydroxide is 1: 1-1: 10, and the volume ratio of dioxane to water is 1: 1-20: 1; dropwise adding the lithium hydroxide aqueous solution into the intermediate IV mixed solution, and continuously stirring at room temperature for reaction for 4-6 h; monitoring the reaction to be complete through TLC, cooling to room temperature, distilling under reduced pressure to remove dioxane in the system, and adjusting the pH to 3-4 with a dilute hydrochloric acid solution to separate out a solid; and then carrying out suction filtration, ice water washing and drying treatment to obtain the intermediate V.

Based on the above preparation method, the preparation of formula VI comprises the steps of: dissolving the intermediate V and HATU in DMF according to a molar ratio of 1: 1-1: 3, and stirring at normal temperature for 1-48 h to obtain the system S1; simultaneously reacting amine compound R⁴Dissolving H and an organic base DIPEA in DMF according to a molar ratio of 1: 1-1: 3, and stirring at normal temperature for 0.5-2H to obtain a system S2; dropwise adding the system S2 into the system S1, and continuously reacting at normal temperature for 6-8 hours; after completion of the reaction, monitored by TLC, a product mixture was formed; adding equal amount of water and ethyl acetate to the product mixture for extraction, extracting the aqueous phase three times with ethyl acetate, and combining the organic layers; and drying the organic layer by anhydrous magnesium sulfate, performing suction filtration, adding silica gel for stir-frying, and performing column chromatography separation to obtain the product of the general formula VI.

Based on the preparation method, the method also comprises the step of preparing the general formula VII, dissolving the general formula VI in ethanol and dropwise adding concentrated hydrochloric acid to react at room temperature to obtain the hydrochloride product of the general formula VI, namely the general formula VII. Specifically, dissolving the intermediate VI in ethanol, and stirring at normal temperature to form an intermediate VI solution with the molar concentration of 0.1-10 mmol/mL; dropwise adding 2-3 drops of concentrated hydrochloric acid into the intermediate VI solution, and continuously stirring for 6-8 hours; and after the reaction is completely monitored by TLC, carrying out reduced pressure distillation to remove the solvent, adding ethyl acetate into the residue to separate out the product, carrying out suction filtration, and washing the filter cake with ethyl ester to obtain the hydrochloride product with the general formula VII.

Therefore, the reaction process of the difluorophenyl cyclopropylamine compound provided by the invention is as follows:

the invention also provides application of the difluorophenyl cyclopropylamine compound in preparation of an LSD1 inhibitor.

Therefore, the difluorophenyl cyclopropylamine compound provided by the scheme is a 3, 4-difluorophenyl cyclopropylamine derivative or a pharmaceutically acceptable salt thereof, and experiments prove that the difluorophenyl cyclopropylamine compound has the effect of inhibiting LSD1, so that a new way is developed for searching a new antitumor drug based on an LSD1 target.

In addition, the invention also provides a preparation method of the 3, 4-difluorophenyl cyclopropylamine compound, which comprises the steps of sequentially and respectively preparing an intermediate IV, an intermediate V, an intermediate VI and an intermediate VII, wherein the intermediate IV is prepared by adopting a three-raw-material one-pot reaction method, pre-activating 3, 4-difluorophenyl cyclopropylamine and ethyl glyoxylate in a mixed solvent of HFIP and toluene to form an imine intermediate, adding an indole compound III, and directly combining three molecules into one molecule through an addition reaction promoted by HFIP to prepare the intermediate IV, the preparation method does not need transition metal catalysis, does not need high-temperature heating and inert gas protection, and is simple in preparation method and simplified in the step of preparing the 3, 4-difluorophenyl cyclopropylamine compound; meanwhile, the substrate has wide application range and high yield which can reach 85 percent.

Detailed Description

The technical solution of the present invention is further described in detail by the following embodiments. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art.

Example 1

This example provides a compound VI-1 having the formula:

the synthetic scheme of compound VI-1 is shown below:

specifically, this example also provides a method for preparing compound VI-1, including the following steps:

preparation of intermediate IV-1: 2.53g, about 15mmol of the starting material 3, 4-difluorophenylalanylamine I and 1.02g, about 10mmol of ethyl glyoxylate II were charged into a round-bottomed flask, and then a mixed solvent consisting of 12mL of toluene and 4mL of hexafluoroisopropanol was added thereto; after stirring at normal temperature for about 30min, 1.17g and about 10mmol of indole III-1 are added into the reaction system and stirring is continued for 6h at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then silica gel was added to fry the sample, and column chromatography was performed to obtain 2.5g of yellow oily IV-1 in 68% yield.

Preparation of intermediate V-1: 3.7g of about 10mmol of intermediate IV-1 are dissolved in 15mL of dioxane and stirred at 0 ℃ in an ice bath, 957mg of about 40mmol of lithium hydroxide are then dissolved in 10mL of water and added dropwise to the reaction system, and the reaction is continued for 6h at room temperature. And monitoring the reaction to be complete by TLC, cooling the system to room temperature, distilling under reduced pressure to remove dioxane in the system, and adjusting the pH to 3-4 by using 1M dilute hydrochloric acid solution to precipitate a solid. And (3) carrying out suction filtration on the reaction system, washing a filter cake with a small amount of ice water, and drying to obtain 3.4g of white solid V-1 with the yield of 99%.

Preparation of product VI-1: 342mg, about 1mmol of intermediate V-1 and 574mg, about 1.5mmol of HATU are dissolved in DMF and stirred at normal temperature for 1h to obtain system S1, and 140mg, about 1.5mmol of aniline and 195mg, about 1.5mmol of organic base DIPEA are dissolved in DMF and stirred at normal temperature for 1h to obtain system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 8 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 50mg yellow solid VI-1 with yield of 12%;¹H NMR(400MHz,DMSO-d₆) δ 11.03-10.93(m,1H),10.10-9.97(m,1H),7.82-7.68(m,1H),7.67-7.57(m,2H),7.39-7.15(m,5H),7.12-6.92(m,4H),6.90-6.76(m,1H),4.75(s,1H),3.25(s,1H),2.32-2.20(m,1H),2.05-1.90(m,1H),1.21-1.11(m,1H),1.10-0.98(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-1 provided in this example is shown above.

Example 2

This example provides a compound VI-2 having the formula:

this example also provides a method for preparing compound VI-2, comprising the steps of:

preparation of product VI-2: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 160mg of benzylamine (about 1.5mmol) and 195mg of DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 8 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, vacuum filtering, adding silica gel, parching, and separating by column chromatography to obtain57mg of VI-2 as a yellow solid in 13% yield;¹H NMR(400MHz,DMSO-d₆) δ 10.97 to 10.87(m,1H),8.61 to 8.48(m,1H),7.77 to 7.61(m,1H),7.33(t, J ═ 7.5Hz,1H),7.28 to 7.15(m,6H),7.13 to 7.01(m,2H),7.01 to 6.89(m,2H),6.85 to 6.77(m,1H),4.61(d, J ═ 5.4Hz,1H),4.38 to 4.14(m,2H),3.05(s,1H),2.28 to 2.17(m,1H),2.03 to 1.85(m,1H),1.14 to 1.05(m,1H),1.04 to 0.94(t, J ═ 5.8Hz, 1H); therefore, it can be confirmed that the structural formula of the compound VI-2 provided in this example is shown above.

Example 3

This example provides a compound VI-3 having the formula:

this example also provides a method for preparing compound VI-3, comprising the steps of:

preparation of product VI-3: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 181mg of p-methylbenzylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 8 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, performing suction filtration, adding silica gel, parching, and performing column chromatography separation to obtain 49mg of yellow solid VI-3 with a yield of 11%;¹H NMR(400MHz,DMSO-d₆) δ 10.92(d, J ═ 7.5Hz,1H),8.57-8.44(m,1H),8.06(t, J ═ 5.4Hz,1H),7.77-7.60(m,1H),7.33(t, J ═ 7.4Hz,1H),7.27-7.24(m,2H),7.22-7.19(m,2H),7.10(d, J ═ 7.8Hz,1H),7.06-7.03(m,1H),6.99(s,1H),6.96-6.90(m,1H),6.86-6.76(m,1H),4.60(d, J ═ 7.6Hz,1H),4.33-4.26(m,2H),3.05(s,1H),2.30(s,2H),2.26 (m,2H), 1H, and 1H, 7, 2, 7, 2, 7, 2, 7; therefore, it can be confirmed that the structural formula of the compound VI-3 provided in this example is as shown above.

Example 4

This example provides a compound VI-4 having the formula:

this example also provides a method for preparing compound VI-4, comprising the steps of:

preparation of product VI-4: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 205mg of p-methoxybenzylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 48mg yellow solid VI-4 with a yield of 10%;¹H NMR(400MHz,DMSO-d₆) δ 10.91(d, J ═ 7.2Hz,1H),8.53-8.43(m,1H),7.77-7.59(m,1H),7.38-7.26(m,2H),7.25-7.18(m,1H),7.18-7.10(m,2H),7.09-7.01(m,2H),7.00-6.89(m,2H),6.84-6.70(m,1H),4.58(d, J ═ 8.3Hz,1H),4.32-4.20(m,2H),3.75(s,1H),3.70(s,3H),3.04(s,1H),2.27-2.14(m,1H),2.09-1.81(m,1H),1.14-1.05(m,1H),1.03-0.93(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-4 provided in this example is as shown above.

Example 5

This example provides a compound VI-5 having the formula:

this example also provides a method for preparing compound VI-5, comprising the steps of:

preparation of product VI-5: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, as275mg of benzhydrylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 55mg yellow solid VI-5 with yield of 11%;¹H NMR(400MHz,DMSO-d₆) δ 10.89(s,1H),8.95(t, J ═ 7.9Hz,1H),7.76-7.61(m,1H),7.47-7.34(m,2H),7.33-7.27(m,3H),7.25-7.18(m,5H),7.17-7.09(m,3H),7.05(t, J ═ 7.6Hz,1H),6.99-6.64(m,3H),6.23-6.09(m,1H),4.81-4.70(m,1H),3.11(s,1H),2.25-2.15(m,1H),2.01-1.88(m,1H),1.15-1.03(m,1H),1.03-0.93(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-5 provided in this example is as shown above.

Example 6

This example provides a compound VI-6 having the formula:

this example also provides a method for preparing compound VI-6, comprising the steps of:

preparation of product VI-6: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 198mg of p-cyanobenzylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 57mg yellow solid VI-6 with yield of 13%;¹H NMR(400MHz,DMSO-d₆)δ10.98(d,J＝7.5Hz,1H),8.71(t,J＝6.1Hz,1H),7.84-7.79(m,1H),7.68(d,J＝8.2Hz1H),7.64(d, J ═ 7.9Hz,1H),7.54 to 7.49(m,1H),7.38(d, J ═ 8.0Hz,2H),7.34(d, J ═ 8.0Hz,1H),7.26 to 7.22(m,1H),7.18(d, J ═ 2.4Hz,1H),7.09 to 7.04(m,1H),6.96 to 6.91(m,1H),6.84 to 6.77(m,1H),4.66(s,1H),4.41 to 4.36(m,2H),2.89(s,1H),2.25 to 2.17(m,1H),2.05 to 1.95(m,1H),1.14 to 1.07(m,1H),1.04 to 0.97(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-6 provided in this example is as shown above.

Example 7

This example provides a compound VI-7 having the formula:

this example also provides a method for preparing compound VI-7, comprising the steps of:

preparation of product VI-7: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 187mg of p-fluorobenzylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 62mg yellow solid VI-7 with a yield of 14%;¹H NMR(400MHz,DMSO-d₆) δ 10.82(d, J ═ 7.5Hz,1H),8.61-8.53(m,1H),8.07(s,1H),7.75-7.59(m,1H),7.46-7.40(m,2H),7.33(t, J ═ 7.7Hz,1H),7.26-7.24(m,1H),7.22-7.19(m,1H),7.17-7.13(m,1H),7.07-7.04(m,1H),7.01-6.94(m,2H),6.85-6.75(m,1H),4.59(d, J ═ 5.3Hz,1H),4.34(d, J ═ 4.3Hz,2H),2.89(s,1H),2.24-2.15(m,1H),2.00 (m,1H), 1.95-1H), 1.06(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-7 provided in this example is as shown above.

Example 8

This example provides a compound VI-8 having the formula:

this example also provides a method for preparing compound VI-8, comprising the steps of:

preparation of product VI-8: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 181mg of N-methylbenzylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 61mg yellow solid VI-8 with a yield of 14%;¹H NMR(400MHz,DMSO-d₆) δ 10.99(d, J ═ 7.5Hz,1H),7.80-7.51(m,1H),7.39-7.33(m,1H),7.33-7.26(m,1H),7.25-7.20(m,2H),7.18-7.11(m,2H),7.11-7.02(m,2H),6.99(t, J ═ 7.6Hz,2H),6.92(d, J ═ 7.4Hz,1H),6.87-6.75(m,1H),5.10-4.88(m,1H),4.77-4.56(m,1H),4.53-4.05(m,1H),3.04(s,1H),2.86-2.75(m,3H),2.30-2.09(m,1H), 2.80-1H), 15.0 (m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-8 provided in this example is as shown above.

Example 9

This example provides a compound VI-9 having the formula:

this example also provides a method for preparing compound VI-9, comprising the steps of:

preparation of product VI-9: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1 while 150mg of N-methylpiperazine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMFDMF was stirred at room temperature for 1 hour to obtain system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 76mg yellow solid VI-9 with a yield of 18%;¹H NMR(400MHz,DMSO-d₆) δ 11.00-10.97(m,1H),7.75-7.57(m,1H),7.33(t, J ═ 9.2Hz,1H),7.28-7.21(m,1H),7.21-7.11(m,1H),7.10-7.01(m,2H),7.00-6.88(m,1H),6.87-6.77(m,1H),5.08-4.98(m,1H),3.69-3.42(m,3H),3.29-3.00(m,2H),2.29-2.08(m,3H),2.07-1.97(m,3H),1.97-1.75(m,3H),1.18-1.04(m,1H),1.03-0.94(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-9 provided in this example is as shown above.

Example 10

This example provides a compound VI-10 having the formula:

this example also provides a method for preparing compound VI-10, comprising the steps of:

preparation of product VI-10: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 171mg of N-ethylpiperazine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 66mg yellow solid VI-10 with yield of 15%;¹H NMR(400MHz,DMSO-d₆)δ10.99-10.89(m,1H),7.74-7.55(m,1H),7.33(t,J＝9.2Hz,1H),7.25(t,J＝9.3Hz,1H),7.19-7.09(m,1H),7.09-6.99(m,2H),6.99-6.87(m,1H),6.87-6.79(m,1H)5.07-4.94(m,1H),3.75-3.38(m,3H),3.29-3.08(m,2H),2.95(s,1H),2.36-2.24(m,1H),2.20-2.07(m,3H),2.04-1.88(m,2H),1.87-1.74(m,1H),1.12-1.02(m,1H),1.02-0.95(m,1H),0.93-0.81(m, 3H); therefore, it can be confirmed that the structural formula of the compound VI-10 provided in this example is shown above.

Example 11

This example provides a compound VI-11 having the formula:

this example also provides a method for preparing compound VI-11, comprising the steps of:

preparation of product VI-11: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 279mg of N-Boc piperazine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 102mg yellow solid VI-11 with a yield of 20%;¹H NMR(400MHz,DMSO-d₆) δ 11.01-10.91(m,1H),7.73-7.57(m,1H),7.33(t, J ═ 9.4Hz,1H),7.28-7.11(m,2H),7.10-6.99(m,2H),6.98-6.89(m,1H),6.88-6.74(m,1H),5.02(s,1H),3.55-3.46(m,2H),3.28-3.08(m,3H),2.93-2.84(m,4H),2.14(s,1H),1.91(s,1H),1.42(s,3H),1.34(s,6H),1.16-1.03(m,1H),1.03-0.91(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-11 provided in this example is shown above.

Example 12

This example provides a compound VI-12 having the formula:

this example also provides a method for preparing compound VI-12, comprising the steps of:

preparation of product VI-12: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 192mg of N-acetylpiperazine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 44mg yellow solid VI-12 with a yield of 10%;¹H NMR(400MHz,DMSO-d₆) δ 11.03-10.92(m,1H),7.64-7.56(m,1H),7.36-7.30(m,1H),7.27-7.20(m,1H),7.18-7.11(m,1H),7.09-7.00(m,2H),6.91(t, J ═ 7.6Hz,1H),6.88-6.76(m,1H),5.04(s,1H),3.70-3.46(m,4H),3.42-3.37(m,1H),3.30-3.19(m,3H),2.13(s,1H),2.00-1.82(m,5H),1.12-1.06(m,1H),1.03-0.94(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-12 provided in this example is shown above.

Example 13

This example provides a compound VI-13 having the formula:

this example also provides a method for preparing compound VI-13, comprising the steps of:

preparation of product VI-13: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 288mg of 1- (4-methoxyphenyl) piperazine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of acetic acid were added to the systemThe ethyl ester was extracted, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 43mg yellow solid VI-13 with yield of 8%;¹H NMR(400MHz,DMSO-d₆) δ 11.00-10.90(m,1H),7.76-7.58(m,1H),7.37-7.28(m,1H),7.25-7.10(m,2H),7.09-6.99(m,2H),6.99-6.88(m,1H),6.87-6.80(m,1H),6.79-6.70(m,4H),5.14-5.00(m,1H),3.80-3.56(m,6H),3.53-3.38(m,1H),3.00-2.67(m,5H),2.23-2.10(m,1H),2.03-1.88(m,1H),1.18-1.04(m,1H),1.02-0.94(m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-13 provided in this example is shown above.

Example 14

This example provides a compound VI-14 having the formula:

this example also provides a method for preparing compound VI-14, comprising the steps of:

preparation of product VI-14: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 195mg of 1- (2-hydroxyethyl) piperazine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 55mg yellow solid VI-14 with yield of 12%;¹H NMR(400MHz,DMSO-d₆) δ 11.00-10.91(m,1H),7.73-7.56(m,1H),7.37-7.30(m,1H),7.28-7.21(m,1H),7.20-7.10(m,1H),7.10-7.01(m,2H),7.00-6.87(m,1H),6.86-6.78(m,1H),5.07-4.98(m,1H),4.41-4.33(m,1H),3.67-3.40(m,5H),3.28-3.11(m,2H),2.35-2.09(m,6H),1.96-1.84(m,2H),1.13-1.04(m,1H),1.03-0.95(m, 1H); thus, the quantization provided by the present embodiment can be confirmedThe structural formula of the compound VI-14 is shown as above.

Example 15

This example provides a compound VI-15 having the formula:

this example also provides a method for preparing compound VI-15, comprising the steps of:

preparation of product VI-15: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 348mg of 1- (4-tert-butylbenzyl) piperazine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 51mg yellow solid VI-15 with yield of 9%;¹H NMR(400MHz,DMSO-d₆) δ 10.99 to 10.92(m,1H),7.73 to 7.57(m,1H),7.34(t, J ═ 8.5Hz,1H),7.30(d, J ═ 8.1Hz,2H),7.27 to 7.21(m,1H),7.20 to 7.14(m,1H),7.14 to 7.09(m,2H),7.08 to 7.01(m,2H),6.99 to 6.88(m,1H),6.86 to 6.79(m,1H),5.07 to 4.99(m,1H),3.74 to 3.54(m,1H),3.54 to 3.43(m,2H),3.32 to 3.24(m,2H),3.23 to 3.11(m,2H),2.33 to 2.24(m,1H), 2.3.23 to 3.11(m,2H),2.33 to 2.24(m,1H), 2.04 to 2.90 (m,1H), 1H, 1.13 to 1H), 1H, 3.3.3.3.3.3.3.3, 3.3.3, 3, 1, 3, 1, 3, 1, 3, 1, 3, 1, 3, 1, H, 1, H, 1, 1H) (ii) a Therefore, it can be confirmed that the structural formula of the compound VI-15 provided in this example is shown above.

Example 16

This example provides a compound VI-16 having the formula:

this example also provides a method for preparing compound VI-16, comprising the steps of:

preparation of product VI-16: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 149mg of 4-piperidone (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 45mg yellow solid VI-16 with yield of 11%;¹H NMR(400MHz,DMSO-d₆) δ 10.95-10.86(m,1H),8.15-8.07(m,1H),7.76-7.61(m,1H),7.37-7.29(m,1H),7.28-7.22(m,1H),7.22-7.15(m,1H),7.10-7.00(m,2H),7.00-6.89(m,1H),6.89-6.82(m,1H),4.61-4.53(m,1H),4.15-4.00(m,1H),3.03(s,1H),2.45-2.29(m,2H),2.27-2.12(m,3H),2.03-1.84(m,3H),1.73-1.55(m,2H),1.16-1.07(m,1H), 1.96-0.07 (m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-16 provided in this example is shown above.

Example 17

This example provides a compound VI-17 having the formula:

this example also provides a method for preparing compound VI-17, comprising the steps of:

preparation of product VI-17: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 106mg of cyclopropylmethylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Organic layer with anhydrous sulfurDrying magnesium, vacuum filtering, adding silica gel, parching, and separating by column chromatography to obtain yellow solid VI-17 (46 mg), with yield of 12%;¹H NMR(400MHz,DMSO-d₆) δ 10.94-10.85(m,1H),8.17-8.05(m,1H),7.74-7.62(m,1H),7.37-7.23(m,2H),7.23-7.14(m,1H),7.11-6.99(m,2H),6.99-6.89(m,1H),6.88-6.79(m,1H),4.53(s,1H),3.11-2.95(m,2H),2.94-2.84(m,1H),2.21(s,1H),2.05-1.84(m,1H),1.18-0.96(m,3H),0.94-0.78(m,1H),0.54-0.26(m,2H),0.25-0.05(m, 2H); therefore, it can be confirmed that the structural formula of the compound VI-17 provided in this example is shown above.

Example 18

This example provides a compound VI-18 having the formula:

this example also provides a process for preparing compound VI-18, comprising the steps of:

preparation of product VI-18: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 176mg of 3-isopropoxypropylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 47mg yellow solid VI-18 with yield of 11%;¹H NMR(400MHz,DMSO-d₆) δ 10.95-10.87(m,1H),8.06-7.98(m,1H),7.75-7.61(m,1H),7.33(t, J ═ 7.6Hz,1H),7.29-7.21(m,1H),7.20-7.15(m,1H),7.11-6.99(m,2H),6.99-6.90(m,1H),6.89-6.78(m,1H),4.51(s,1H),3.45-3.36(m,1H),3.31-3.19(m,2H),3.18-3.08(m,2H),2.24-2.16(m,1H),2.04-1.84(m,1H),1.65-1.48(m,2H),1.14-0.92(m, 9H); therefore, it can be confirmed that the structural formula of the compound VI-18 provided in this example is shown above.

Example 19

This example provides a compound VI-19 having the formula:

this example also provides a method for preparing compound VI-19, comprising the steps of:

preparation of product VI-19: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 153mg of N, N-dimethylpropanediamine (about 1.5mmol) and 195mg of DIPEA (about 1.5mmol) as an organic base were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 64mg yellow solid VI-19 with yield of 15%;¹H NMR(400MHz,DMSO-d₆) δ 10.98-10.89(m,1H),8.08(t, J ═ 5.6Hz,1H),7.64(d, J ═ 8.0Hz,1H),7.35-7.30(m,1H),7.28-7.22(m,1H),7.18-7.14(m,1H),7.07-7.02(m,1H),7.02-6.95(m,1H),6.93(t, J ═ 7.5Hz,1H),6.89-6.83(m,1H),4.50(s,1H),3.15-3.04(m,2H),2.23-2.07(m,5H),2.06-2.00(m,6H),1.56-1.44(m,2H),1.13-1.05(m,1H), 1.95-1H (m, 1H); therefore, it can be confirmed that the structural formula of the compound VI-19 provided in this example is shown above.

Example 20

This example provides a compound VI-20 having the formula:

this example also provides a method for preparing compound VI-20, comprising the steps of:

preparation of product VI-20: 342mg of intermediate V-1 (ca. 1mmol) and 574mg of HATU (ca. 1.5 m)mol) in DMF and stirred at room temperature for 1h to obtain system S1, while 110mg of diethylamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) are dissolved in DMF and stirred at room temperature for 1h to obtain system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. Drying the organic layer with anhydrous magnesium sulfate, filtering, adding silica gel, parching, and separating by column chromatography to obtain 40mg yellow solid VI-20 with a yield of 10%;¹H NMR(400MHz,DMSO-d₆) δ 11.00-10.91(m,1H),7.76-7.59(m,1H),7.36-7.31(m,1H),7.28-7.21(m,1H),7.16-7.10(m,1H),7.08-7.06(m,1H),7.04-6.99(m,1H),6.98-6.88(m,1H),6.88-6.80(m,1H),4.87(s,1H),3.52-3.44(m,1H),3.31-3.24(m,1H),3.23-3.05(m,3H),2.19-2.10(m,1H),2.00-1.91(m,1H),1.03-0.97(m,3H),0.96-0.90(m,3H),0.81-0.68(m, 0H); therefore, it can be confirmed that the structural formula of the compound VI-20 provided in this example is shown above.

Example 21

This example provides a compound VII-1 having the formula:

this example also provides a method for preparing compound VII-1, comprising the steps of:

preparation of product VII-1: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 192mg of 4-dimethylaminopiperidine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered under suction, and then, silica gel was added thereto to fry the sample, and column chromatography was performed to obtain 66mg of a yellow oil with a yield of 15%. The oil was dissolved in ethanol and the mixture was washed with water,adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and removing the solvent by reduced pressure distillation to obtain a hydrochloride product VII-1;¹H NMR(400MHz,DMSO-d₆) δ 11.87-11.66(m,1H),11.42-10.98(m,1H),10.35-9.33(m,2H),7.83-7.68(m,1H),7.65-7.53(m,1H),7.52-7.40(m,1H),7.38-7.27(m,1H),7.27-7.17(m,1H),7.17-7.05(m,1H),7.05-6.97(m,1H),6.96-6.54(m,1H),6.07(s,1H),4.65-4.49(m,1H),4.20-3.90(m,2H),3.37-3.25(m,1H),3.15-2.96(m,1H),2.81-2.56(m,6H), 2.01-2.32 (m,1H), 1.01-1H), 1.51-1H, 1.31-4.31H, 1H) (ii) a Therefore, it can be confirmed that the structural formula of the compound VII-1 provided in this example is as shown above.

Example 22

This example provides a compound VII-2 having the formula:

this example also provides a method for preparing compound VII-2, comprising the steps of:

preparation of product VII-2: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 36mg of a yellow oil with a yield of 9%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-2;¹H NMR(400MHz,DMSO-d₆)δ11.58-11.50(,1H),10.39(s,1H),10.35-9.57(m,2H),9.04-8.94(m,1H),7.80-7.57(m,1H),7.56-7.45(m,1H),7.45-7.28(m,2H),7.21-7.07(m,2H),7.05-6.92(m,1H),6.92-6.56(m,1H),5.55-5.33(m,1H),3.75-3.63(m,1H) 3.39(s,2H),3.31-3.20(m,2H),3.11(s,1H),2.81-2.61(m,6H),1.66-1.42(m,1H),1.34-1.21(m, 1H); therefore, it can be confirmed that the structural formula of the compound VII-2 provided in this example is as shown above.

Example 23

This example provides a compound VII-3 having the formula:

this example also provides a method for preparing compound VII-3, comprising the steps of:

preparation of product VII-3: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 153mg of N, N, N' -trimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 40mg of a yellow oil with a yield of 9%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-3;¹H NMR(400MHz,DMSO-d₆) δ 11.66-11.56(m,1H),10.37-10.00(m,1H),10.00-9.78(m,1H),9.77-9.53(m,1H),7.80-7.60(m,1H),7.60-7.49(m,1H),7.47-7.38(m,1H),7.38-7.25(m,1H),7.23-7.09(m,2H),7.07-6.90(m,2H),5.78(s,1H),4.32-4.11(m,1H),3.25-3.13(m,2H),2.95-2.89(m,3H),2.88-2.82(m,3H),2.81-2.74(m,3H),2.70-2.56(m,3H), 1.44-1H), 1.14-1H (m, 1H); therefore, it can be confirmed that the structural formula of the compound VII-3 provided in this example is as shown above.

Example 24

This example provides a compound VII-4 having the formula:

this example also provides a method for preparing compound VII-4, comprising the steps of:

preparation of product VII-4: 342mg of intermediate V-1 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 90mg of ethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered under suction, and then, silica gel was added thereto, and the resulting mixture was subjected to column chromatography to obtain 38mg of a yellow oil in a yield of 10%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-4;¹H NMR(400MHz,DMSO-d₆) δ 11.56-11.48(m,1H),10.43-10.13(m,1H),9.94-9.57(m,1H),8.94(t, J ═ 5.7Hz,1H),8.18(s,3H),7.80-7.58(m,1H),7.56-7.45(m,1H),7.45-7.26(m,2H),7.22-7.08(m,2H),7.06-6.94(m,1H),6.94-6.54(m,1H),5.49-5.29(m,1H),3.61-3.52(m,1H),3.20-3.09(m,1H),2.98-2.88(m,1H),2.87-2.76(m,1H),2.66-2.20(m, 2.20), 1.36H, 1H, 36H, 1H, 6, 1H, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1, 6, 1; therefore, it can be confirmed that the structural formula of the compound VII-4 provided in this example is as shown above.

Example 25

This example provides a compound VII-5 having the formula:

this example also provides a method for preparing compound VII-5, comprising the steps of:

preparation of intermediate IV-2: the preparation of intermediate IV-2 provided in this example and that provided in example 1The preparation method of the intermediate IV-1 is basically the same, and the main difference is that: 1.31g and about 10mmol of indole compound III-2 are added into the reaction system, and stirring is continued for 6 hours at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then a silica gel stir-fried sample was added and separated by column chromatography to obtain 3.26g of yellow oily IV-2 with a yield of 85%. Wherein the structural formula of the indole compound III-2 is

The structural formula of the intermediate IV-2 is

Preparation of intermediate V-2: the preparation method of the intermediate IIV-2 provided in this example is substantially the same as the preparation method of the intermediate V-1 provided in example 1, with the main difference that: reaction was carried out using 3.84g of about 10mmol of intermediate IV-2 to give 3.55g of white solid V-2 in 99% yield. Wherein the intermediate V-2 has a structural formula

Preparation of product VII-5: 356mg of intermediate V-2 (ca. 1mmol) and 574mg of HATU (ca. 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (ca. 1.5mmol) and 195mg of the organic base DIPEA (ca. 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 47mg of a yellow oil with a yield of 11%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-5;¹H NMR(400MHz,DMSO-d₆)δ10.37(s,1H),10.36-9.64(m,2H),9.07-8.94(m,1H),7.80-7.56(m,1H),7.52-7.43(m,2H),7.39-7.28(m,1H),7.24-7.17(m,1H),7.16-7.05(m,1H),6.99-6.49(m,2H),5.58-5.35(m,1H),3.79-3.72(m,3H),3.71-3.60(m,1H),3.32-3.17(m,2H),3.16-3.06(m,1H),2.82-2.70(m,6H),2.65-2.10(m,2H),1.65-1.43(m,1H),1.34-1.21(m, 1H); therefore, it can be confirmed that the structural formula of the compound VII-5 provided in this example is as shown above.

Example 26

This example provides a compound VII-6 having the formula:

this example also provides a method for preparing compound VII-6, comprising the steps of:

preparation of intermediate IV-3: the preparation method of intermediate IV-3 provided in this example is substantially the same as the preparation method of intermediate IV-1 provided in example 1, with the main difference that: 1.57g and about 10mmol of indole compound III-3 are added into the reaction system, and stirring is continued for 6 hours at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then a silica gel stir-fried sample was added and separated by column chromatography to obtain 3.28g of yellow oily IV-3 with a yield of 80%. Wherein the structural formula of the indole compound III-3 is

The structural formula of the intermediate IV-3 is

Preparation of intermediate V-3: the preparation method of intermediate IV-3 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction with 4.1g, about 10mmol of intermediate IV-3 gave 3.82g of white solid V-3 in 100% yield. Wherein the intermediate V-3 has a structural formula

Preparation of product VII-6: 382mg of intermediate V-3 (ca. 1mmol) and 574mg of HATU (ca. 1.5 m)mol) in DMF, stirring at normal temperature for 1h to obtain system S1, simultaneously dissolving 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of DIPEA (about 1.5mmol) in DMF, and stirring at normal temperature for 1h to obtain system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered under suction, and then, silica gel was added thereto, and the resulting mixture was subjected to column chromatography to obtain 46mg of a yellow oil with a yield of 10%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-6;¹H NMR(400MHz,DMSO-d₆) δ 10.59-10.16(m,2H),10.04-9.56(m,1H),9.09-8.99(m,1H),7.82-7.63(m,1H),7.58-7.52(m,1H),7.52-7.41(m,1H),7.36-7.11(m,3H),7.11-6.96(m,1H),6.71-6.42(m,1H),5.56-5.30(m,1H),3.74-3.61(m,1H),3.43-3.32(m,1H),3.31-3.16(m,2H),3.15-3.05(m,1H),2.87-2.62(m,7H),2.62-2.53(m,1H),1.63-1.47(m,1H), 1.15-3.05 (m,1H),2.87-2.62(m,7H),2.62-2.53(m,1H), 1.63-1H), 1.47(m,1H), 0.84-0.84 (m, 1H); therefore, it can be confirmed that the structural formula of the compound VII-6 provided in this example is as shown above.

Example 27

This example provides a compound VII-7 having the formula:

this example also provides a method for preparing compound VII-7, comprising the steps of:

preparation of intermediate IV-4: the preparation method of intermediate IV-4 provided in this example is substantially the same as the preparation method of intermediate IV-1 provided in example 1, with the main difference that: 1.93g and about 10mmol of indole compound III-4 are added into the reaction system, and stirring is continued for 6 hours at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then a silica gel stir-fried sample was added and separated by column chromatography to obtain 3.35g of yellow oily IV-4 with a yield of 75%. Wherein the structural formula of the indole compound III-4 is

The structural formula of the intermediate IV-4 is

Preparation of intermediate V-4: the preparation method of intermediate IV-4 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction with 4.46g, about 10mmol of intermediate IV-4 gave 3.89g of white solid V-4 in 93% yield. Wherein the intermediate V-4 has a structural formula

Preparation of product VII-7: 418mg of intermediate V-4 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered under suction, and then, silica gel was added thereto, and the resulting mixture was subjected to column chromatography to obtain 52mg of a yellow oil in a yield of 11%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-7;¹H NMR(400MHz,DMSO-d₆) δ 10.66-10.26(m,2H),10.23-9.63(m,1H),9.12-9.02(m,1H),7.92-7.80(m,1H),7.79-7.69(m,1H),7.63-7.50(m,3H),7.50-7.38(m,3H),7.36-7.27(m,1H),7.27-7.19(m,1H),7.16-6.94(m,2H),6.64-6.40(m,1H),5.70-5.44(m,1H),3.76-3.65(m,1H),3.29-3.17(m,2H),3.16-3.06(m,1H),2.81-2.70(m,7H),2.65-2.04(m,1H), 1.51-1H (m, 1H); therefore, it can be confirmed that the structural formula of the compound VII-7 provided in this example is as shown above.

Example 28

This example provides a compound VII-8 having the formula:

this example also provides a method for preparing compound VII-8, comprising the steps of:

preparation of intermediate IV-5: the preparation method of intermediate IV-5 provided in this example is substantially the same as the preparation method of intermediate IV-1 provided in example 1, with the main difference that: 1.31g and about 10mmol of indole compound III-5 are added into the reaction system, and stirring is continued for 6 hours at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then silica gel was added to fry the sample, and column chromatography separation was carried out to obtain 2.96g of yellow oily IV-5 in 77% yield. Wherein the structural formula of the indole compound III-5 is

The structural formula of the intermediate IV-5 is

Preparation of intermediate V-5: the preparation method of intermediate IV-5 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction with 3.84g, about 10mmol of intermediate IV-5 gave 3.42g of white solid V-5 in 96% yield. Wherein the intermediate V-5 has a structural formula

Preparation of product VII-8: 356mg of intermediate V-5 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, the same amount of water and 20mL of ethyl acetate were added to the system to conduct extractionThe aqueous phase was extracted three times with ethyl acetate and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 55mg of a yellow oil with a yield of 13%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-8;¹H NMR(400MHz,DMSO-d₆) δ 11.49-11.40(m,1H),10.75-10.47(m,1H),10.20-9.80(m,1H),9.76-9.43(m,1H),7.66-7.52(m,1H),7.40-7.31(m,1H),7.30-7.23(m,1H),7.22-7.15(m,1H),7.11-6.99(m,2H),6.99-6.89(m,1H),6.87-6.58(m,1H),5.47-5.27(m,1H),3.76-3.61(m,1H),3.29-3.14(m,1H),3.11-3.02(m,1H),2.75-2.60(m,8H),2.46-2.17(m, 1H), 1.55-1H), 1H (m, 1H); therefore, it can be confirmed that the structural formula of the compound VII-8 provided in this example is as shown above.

Example 29

This example provides a compound VII-9 having the formula:

this example also provides a method for preparing compound VII-9, comprising the steps of:

preparation of intermediate IV-6: the preparation method of intermediate IV-6 provided in this example is substantially the same as the preparation method of intermediate IV-1 provided in example 1, with the main difference that: 1.93g and about 10mmol of indole compound III-6 are added into the reaction system, and stirring is continued for 6 hours at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then a silica gel stir-fried sample was added and separated by column chromatography to obtain 3.7g of yellow oily IV-6 with a yield of 83%. Wherein the structural formula of the indole compound III-6 is

The structural formula of the intermediate IV-6 is

Intermediate V-6The preparation of (1): the preparation method of intermediate IV-6 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction with 4.46g, about 10mmol of intermediate IV-6 gave 3.93g of white solid V-6 in 94% yield. Wherein the intermediate V-6 has a structural formula

Preparation of product VII-29: 418mg of intermediate V-6 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 60mg of a yellow oil with a yield of 12%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-9;¹H NMR(400MHz,DMSO-d₆) δ 12.01-11.86(m,1H),10.82-10.56(m,1H),10.26-9.85(m,1H),9.73-9.46(m,1H),8.72-8.56(m,1H),7.77-7.69(m,1H),7.67-7.58(m,2H),7.56-7.50(m,1H),7.48-7.43(m,1H),7.42-7.34(m,2H),7.27-7.16(m,2H),7.14-7.03(m,1H),6.94-6.70(m,1H),6.70-6.38(m,1H),5.53-5.30(m,1H),3.77-3.68(m,1H),3.36-3.16(m, 3.3H), 3.70-3.38 (m,1H), 3.3.0-6.13.13 (m,1H), 1H) (ii) a Therefore, it can be confirmed that the structural formula of the compound VII-9 provided in this example is as shown above.

Example 30

This example provides a compound VII-10 having the formula:

this example also provides a method for preparing compound VII-10, comprising the steps of:

preparation of intermediate IV-7: the preparation method of intermediate IV-7 provided in this example is substantially the same as the preparation method of intermediate IV-1 provided in example 1, with the main difference that: 2.27g of indole compound III-7 of about 10mmol is added to the reaction system, and stirring is continued for 6h at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then a silica gel stir-fried sample was added and separated by column chromatography to obtain 3.93g of yellow oily IV-7 with a yield of 82%. Wherein the structural formula of the indole compound III-7 is

The structural formula of the intermediate IV-7 is

Preparation of intermediate V-7: the preparation method of intermediate IV-7 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction was carried out using 4.79g of about 10mmol of intermediate IV-7 to give 4.1g of white solid V-7 in 91% yield. Wherein the intermediate V-7 has a structural formula

Preparation of product VII-10: 452mg of intermediate V-7 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 51mg of a yellow oil with a yield of 10%. Dissolving the oily substance in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8 hr, and distilling under reduced pressure to remove solventObtaining a hydrochloride product VII-10;¹H NMR(400MHz,DMSO-d₆) δ 12.05-11.89(m,1H),10.84-10.54(m,1H),10.25-9.40(m,2H),8.68-8.55(m,1H),7.76-7.56(m,4H),7.49-7.41(m,1H),7.39-7.34(m,1H),7.30-7.16(m,2H),7.15-7.04(m,1H),6.94-6.44(m,2H),5.70-5.31(m,1H),3.79-3.69(m,1H),3.40-3.35(m,1H),3.28-3.18(m,2H),3.11-3.02(m,1H),2.79-2.65(m,6H),2.23-1.99(m,1H), 1.96-1H, 0.20-1H); therefore, it can be confirmed that the structural formula of the compound VII-10 provided in this example is as shown above.

Example 31

This example provides a compound VII-11 having the formula:

this example also provides a method for preparing compound VII-11, comprising the steps of:

preparation of intermediate IV-8: the preparation method of intermediate IV-8 provided in this example is substantially the same as the preparation method of intermediate IV-1 provided in example 1, with the main difference that: 1.31g and about 10mmol of indole compound III-8 are added into the reaction system, and stirring is continued for 6 hours at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then silica gel was added to fry the sample, and column chromatography separation was carried out to obtain 2.61g of yellow oily IV-8 with a yield of 68%. Wherein the structural formula of the indole compound III-8 is shown in the specification

The structural formula of the intermediate IV-8 is

Preparation of intermediate V-8: the preparation method of intermediate IV-8 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction with 3.84g, about 10mmol of intermediate IV-8 gave 3.52g of V-8 as a white solid in 99% yield. Wherein, the structure of intermediate V-8Formula (II)

Preparation of product VII-11: 356mg of intermediate V-8 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 41mg of a yellow oil with a yield of 9%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-11;¹H NMR(400MHz,DMSO-d₆) δ 11.46-11.33(m,1H),10.29(s,1H),9.97-9.61(m,1H),8.94-8.83(m,1H),7.53-7.34(m,2H),7.32-7.26(m,1H),7.24-7.11(m,2H),7.06-6.94(m,1H),6.94-6.50(m,2H),5.47-5.27(m,1H),3.75-3.62(m,1H),3.30-3.20(m,2H),3.14-3.05(m,1H),2.84-2.66(m,7H),2.65-2.54(m,1H),2.36-2.16(m,3H),1.61-1.48(m,1H), 1.38-1H (m, 1H); therefore, it can be confirmed that the structural formula of the compound VII-11 provided in this example is as shown above.

Example 32

This example provides a compound VII-12 having the formula:

this example also provides a method for preparing compound VII-12, comprising the steps of:

preparation of intermediate IV-9: the preparation method of intermediate IV-9 provided in this example is substantially the same as the preparation method of intermediate IV-1 provided in example 1, with the main difference that: 1.35g of about 10mmol of indole compound III was added to the reaction system-9 stirring at room temperature for 6 h. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then silica gel was added to fry the sample, and column chromatography separation was carried out to obtain 2.68g of IV-9 as a yellow oil in 69% yield. Wherein the structural formula of the indole compound III-9 is

The structural formula of the intermediate IV-9 is

Preparation of intermediate V-9: the preparation method of intermediate IV-9 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction with 3.88g, about 10mmol of intermediate IV-9 gave 3.6g of V-9 as a white solid in 100% yield. Wherein the intermediate V-9 has a structural formula

Preparation of product VII-12: 360mg of intermediate V-9 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1, while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 58mg of a yellow oil with a yield of 13%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-12;¹H NMR(400MHz,DMSO-d₆) δ 11.75-11.67(m,1H),10.60-10.18(m,2H),10.08-9.67(m,1H),9.17-9.05(m,1H),7.67-7.41(m,2H),7.40-7.22(m,2H),7.22-7.12(m,1H),7.03-6.57(m,2H),5.63-5.28(m,1H),3.74-3.62(m,1H),3.34-3.17(m,2H),3.16-3.05(m,1H),2.86-2.71(m,6H),2.70-2.17(m,2H),1.65-1.47(m,1H),1.34-1.21(m, 1H); thus, it is possible to provideIt can be confirmed that the structural formula of the compound VII-12 provided in this example is shown above.

Example 33

This example provides a compound VII-13 having the formula:

this example also provides a method for preparing compound VII-13, comprising the steps of:

preparation of intermediate IV-10: the preparation method of the intermediate IV-10 provided in this example is substantially the same as the preparation method of the intermediate IV-1 provided in example 1, with the main difference that: 1.96g of indole compound III-10 of about 10mmol is added into the reaction system, and stirring is continued for 6 hours at normal temperature. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and then a silica gel stir-fried sample was added and separated by column chromatography to obtain 3.60g of yellow oily IV-10 with a yield of 80%. Wherein, the structural formula of the indole compound III-10 is

The structural formula of the intermediate IV-10 is

Preparation of intermediate V-10: the preparation method of intermediate IV-10 provided in this example is substantially the same as the preparation method of intermediate V-1 provided in example 1, with the main difference that: reaction was carried out using 4.49g of about 10mmol of intermediate IV-10 to give 4.13g of white solid V-10 in 98% yield. Wherein the intermediate V-10 has a structural formula

Preparation of product VII-13: 421mg of intermediate V-10 (about 1mmol) and 574mg of HATU (about 1.5mmol) were dissolved in DMF and stirred at room temperature for 1h to give system S1 while 132mg of N, N-dimethylethylenediamine (about 1.5mmol) and 195mg of the organic base DIPEA (about 1.5mmol) were dissolved in DMF,stirring at normal temperature for 1h to obtain a system S2. And dropwise adding the system S2 into the system S1, and continuously reacting for 6 hours at normal temperature. After completion of the reaction was monitored by TLC, an equal amount of water and 20mL of ethyl acetate were added to the system for extraction, the aqueous phase was extracted three times with ethyl acetate, and the organic layers were combined. The organic layer was dried over anhydrous magnesium sulfate, filtered, added with silica gel, and subjected to column chromatography to obtain 47mg of a yellow oil with a yield of 10%. Dissolving the oily matter in ethanol, adding 2-3 drops of concentrated hydrochloric acid, stirring at room temperature for 8h, and distilling under reduced pressure to remove the solvent to obtain a hydrochloride product VII-13;¹H NMR(400MHz,DMSO-d₆) δ 11.86-11.74(m,1H),10.37(s,1H),10.21-9.82(m,1H),9.09-8.97(m,1H),7.97-7.69(m,1H),7.59-7.51(m,1H),7.43-7.36(m,1H),7.34-7.26(m,1H),7.25-7.12(m,2H),7.04-6.90(m,1H),6.80-6.50(m,1H),5.53-5.32(m,1H),3.75-3.64(m,1H),3.39-3.37(m,1H),3.32-3.21(m,2H),3.14-3.06(m,1H),2.81-2.71(m, 6.68-1H), 2.65(m, 1H), 1H), 1.46-1H, 1H); therefore, it can be confirmed that the structural formula of the compound VII-13 provided in this example is as shown above.

Assays for inhibition of LSD1 Activity

The experimental method comprises the following steps: the samples were the above-mentioned synthetic compounds VI-1 to VI-20 and VII-1 to VII-13. Sample stock solution: weighing 1-2 mg of samples respectively, dissolving the samples into mother liquor with the concentration of 10mM by using DMSO (dimethyl sulfoxide), and diluting the mother liquor to the concentration to be measured by using DMSO during the experiment. The samples were respectively incubated with human-derived complex protein LSD1/CoREST purified from an E.coli expression system, and then incubated with H3K4me2 polypeptide synthesized by Gill Biochemical (Shanghai) Co., Ltd for 30 min. After the incubation was completed, fluorescent dye Amplex Red and horseradish peroxidase HRP were added to react for 5min, and then a fluorescence signal was measured using an EnVision microplate reader (PerkinElmer, Waltham, MA, USA) (E)_X＝535nm，E_m595nm) and calculating the inhibition ratio. Specifically, the inhibition ratio calculation formula is as follows:

wherein the "fluorescence intensity of the sample group" in the above formula for calculating the inhibition ratioThe degree, the fluorescence intensity of the standard group and the fluorescence intensity of the blank group were measured by the above-mentioned experimental methods, except that: under the same conditions, the object of measurement of "fluorescence intensity of sample group" is a sample liquid containing the above-mentioned compound to be measured, and the object of measurement of "fluorescence intensity of standard group" is a standard liquid containing no sample as compared with the above-mentioned sample liquid. The "blank fluorescence intensity" was measured on a blank sample that did not contain LSD1/CoREST complex protein and H3K4me2 polypeptide. IC of the above sample Compound was treated with Graphpad Prism 8.0₅₀Data, results are shown in table 1.

TABLE 1 data table of inhibitory Activity of the Compounds of the present invention on LSD1/CoREST

Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention and not to limit it; although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art will understand that: modifications to the specific embodiments of the invention or equivalent substitutions for parts of the technical features may be made; without departing from the spirit of the present invention, it is intended to cover all aspects of the invention as defined by the appended claims.

Claims (10)

1. A difluorophenyl cyclopropylamine compound is characterized by being a 3, 4-difluorophenyl cyclopropylamine derivative or a pharmaceutically acceptable salt thereof, wherein the general formula of the 3, 4-difluorophenyl cyclopropylamine derivative is as follows:

wherein the group R in the formula¹Is H, contains C_1～6Alkyl group of (C)_3～7Cycloalkyl, phenyl or substituted phenyl of (a);

radical R²Is H, contains C_1～6Alkyl group of (C)_1～6Halogenoalkyl of (C)_3～7Cycloalkyl of, containing C_3～6Halocycloalkyl, phenyl or substituted phenyl of (a);

radical R³Is H, contains C_1～6Alkyl, phenyl or substituted phenyl, halogen, containing C_1～6Alkoxy, cyano, nitro, trifluoromethyl;

radical R⁴Is hydroxy, containing C_1～6Alkoxy, amino or substituted amino, anilino, benzylamino or substituted benzylamino, piperidinyl, piperazinyl, N-substituted piperazinyl, N-disubstituted amino, C_3～6A cyclic amino group.

2.3, 4-difluorophenypropylamine compound according to claim 1, wherein the group R is¹Is H, methyl, ethyl, propyl, cyclopropyl or phenyl; the group R²Is H, methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, phenyl, or halophenyl; the group R³Is H, 4-methyl, 4-ethyl, 4-propyl, 4-butyl, 4-fluoro, 4-bromo, 3-phenyl, 3-benzyl, 3-methoxy, 3-ethoxy, 3-cyano, 3-nitro, or 4-trifluoromethyl; the group R⁴Is hydroxyl, methoxy, ethoxy, pentoxy, amino, fluoro-amino, methylamino, ethylamino,

3. A difluorophenylpropylamine compound according to claim 2, wherein said 3, 4-difluorophenylpropylamine derivative is one of compounds VI-1 to VI-20 having said general formula VI and the following group:

VI-1:R¹＝H,R²＝H,R³＝H,

VI-2:R¹＝H,R²＝H,R³＝H,

VI-3:R¹＝H,R²＝H,R³＝H,

VI-4:R¹＝H,R²＝H,R³＝H,

VI-5:R¹＝H,R²＝H,R³＝H,

VI-6:R¹＝H,R²＝H,R³＝H,

VI-7:R¹＝H,R²＝H,R³＝H,

VI-8:R¹＝H,R²＝H,R³＝H,

VI-9:R¹＝H,R²＝H,R³＝H,

VI-10:R¹＝H,R²＝H,R³＝H,

VI-11:R¹＝H,R²＝H,R³＝H,

VI-12:R¹＝H,R²＝H,R³＝H,

VI-13:R¹＝H,R²＝H,R³＝H,

VI-14:R¹＝H,R²＝H,R³＝H,

VI-15:R¹＝H,R²＝H,R³＝H,

VI-16:R¹＝H,R²＝H,R³＝H,

VI-17:R¹＝H,R²＝H,R³＝H,

VI-18:R¹＝H,R²＝H,R³＝H,

VI-19:R¹＝H,R²＝H,R³＝H,

VI-20:R¹＝H,R²＝H,R³＝H,

4. a difluorophenylpropylamine compound according to claim 2, wherein said 3, 4-difluorophenylpropylamine derivative is one of compounds VI-1 to VI-13 having said general formula VII and the following groups:

VII-1:R¹＝H,R²＝H,R³＝H,

VII-2:R¹＝H,R²＝H,R³＝H,

VII-3:R¹＝H,R²＝H,R³＝H,

VII-4:R¹＝H,R²＝H,R³＝H,

VII-5:R¹＝CH₃,R²＝H,R³＝H,

VII-6:

R²＝H,R³＝H,

VII-7:

R²＝H,R³＝H,

VII-8:R¹＝H,R²＝CH₃,R³＝H,

VII-9:R¹＝H,

R³＝H,

VII-10:R¹＝H,

R³＝H,

VII-11:R¹＝H,R²＝H,R³＝4-CH₃,

VII-12:R¹＝H,R²＝H,R³＝4-F,

VII-13:R¹＝H,R²＝H,R³＝4-Br,

5. a method for preparing difluorocyclopropylamine compound as claimed in any one of claims 1 to 4, comprising the steps of:

the preparation of the intermediate IV is that 3, 4-difluorophenyl cyclopropylamine and ethyl glyoxylate are dissolved in a toluene mixed solvent and then react with an indole compound III at normal temperature to prepare the intermediate IV; wherein the toluene mixed solvent mainly comprises toluene and hexafluoroisopropanol, and the structural general formula of the indole compound III is

The structural general formula of the intermediate IV is

Preparation of intermediate V the intermediate IV is first reacted with aqueous lithium hydroxide solution in an organic solvent at room temperature; removing the organic solvent, and adjusting the pH value to 3-4 by using hydrochloric acid to precipitate a solid; then carrying out suction filtration and separation to obtain an intermediate V; wherein the organic solvent is tetrahydrofuran or dioxane, and the structural general formula of the intermediate V is

Preparation of general formula VI the intermediates V and HATU were dissolved in DMF at room temperature to form formation system S1; reacting an amine compound R⁴Dissolving H and organic base DIPEA in DMF to form a system S2; the system S2 was added dropwise to the system S1 and reacted at room temperature to produce the product of general formula VI.

6. The process according to claim 5, characterized in that the step of preparing the intermediate IV comprises: uniformly stirring 3, 4-difluorophenylalanyl amine and ethyl glyoxylate with the mixed solvent at normal temperature according to the molar ratio of 1: 1-1: 3, wherein the volume ratio of toluene to hexafluoroisopropanol in the mixed solvent is 1: 1-5: 1; and then adding the substituted indole III, continuing stirring at normal temperature for 6-12 h, monitoring by TLC (thin layer chromatography) to complete reaction, cooling to room temperature, adding silica gel, stir-frying, and performing column chromatography separation to obtain the intermediate IV.

7. The process according to claim 6, characterized in that the step of preparation of intermediate V comprises: dissolving the intermediate IV in dioxane, and stirring in an ice bath at 0 ℃ to form an intermediate IV mixed solution; then dissolving lithium hydroxide in water to form the lithium hydroxide aqueous solution, wherein the molar ratio of the intermediate IV to the lithium hydroxide is 1: 1-1: 10, and the volume ratio of dioxane to water is 1: 1-20: 1; dropwise adding the lithium hydroxide aqueous solution into the intermediate IV mixed solution, and continuously stirring at room temperature for reaction for 4-6 h; monitoring the reaction to be complete through TLC, cooling to room temperature, distilling under reduced pressure to remove dioxane in the system, and adjusting the pH to 3-4 with a dilute hydrochloric acid solution to separate out a solid; and then carrying out suction filtration, ice water washing and drying treatment to obtain the intermediate V.

8. The method of claim 7, wherein the step of preparing of formula VI comprises: dissolving the intermediate V and HATU in DMF according to a molar ratio of 1: 1-1: 3, and stirring at normal temperature for 1-48 h to obtain the system S1; simultaneously reacting amine compound R⁴H anddissolving DIPEA (DiPEA) in DMF (dimethyl formamide) according to a molar ratio of 1: 1-1: 3, and stirring at normal temperature for 0.5-2 h to obtain a system S2; dropwise adding the system S2 into the system S1, and continuously reacting at normal temperature for 6-8 hours; after completion of the reaction, monitored by TLC, a product mixture was formed; adding equal amount of water and ethyl acetate to the product mixture for extraction, extracting the aqueous phase three times with ethyl acetate, and combining the organic layers; and drying the organic layer by anhydrous magnesium sulfate, performing suction filtration, adding silica gel for stir-frying, and performing column chromatography separation to obtain the product of the general formula VI.

9. The preparation method according to any one of claims 5 to 8, further comprising the step of preparing the general formula VII, dissolving the general formula VI in ethanol and dropwise adding concentrated hydrochloric acid to react at room temperature to obtain the hydrochloride product of the general formula VI, namely the general formula VII.

10. Use of difluorocyclopropylamine compounds according to any one of claims 1 to 4 as LSD1 inhibitors.