Preparation method of trans-4-(2-(4-(benzo [b] thiophene-7-yl) piperazine-1-yl) ethyl) cyclohexyl) urea and intermediate of trans-4-(2-(4-(benzo [b] thiophene-7-yl) piperazine-1-yl) ethyl) cyclohexyl) urea

Abstract

The invention provides a preparation method of trans-4-(2-(4-(benzo [b] thiophene-7-yl) piperazine-1-yl) ethyl) cyclohexyl) urea and an intermediate for synthesizing the trans-4-(2-(4-(benzo [b] thiophene-7-yl) piperazine-1-yl) ethyl) cyclohexyl) urea. The invention further provides a preparation method of the trans-4-(2-(4-(benzo [b] thiophene-7-yl) piperazine-1-yl) ethyl) cyclohexyl) urea.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical synthesis, and specifically relates to a method for preparing trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazine-1-yl)ethyl)cyclohexyl)urea, and an intermediate for synthesizing the trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazine-1-yl)ethyl)cyclohexyl)urea. Background Art

[0002] The structure of trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazin-1-yl)ethyl)cyclohexyl)urea is shown in the following formula b:

[0003]

[0004] Chinese patent application (application number: CN202310083508.1) describes trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazin-1-yl)ethyl)cyclohexyl)urea and its preparation method:

[0005]

[0006] However, it is of great significance to research and develop more different synthesis processes and intermediates so as to select processes and intermediates that are more suitable for actual industrial scenarios such as production and transportation in the future. Summary of the invention

[0007] The present invention provides a method for preparing trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazin-1-yl)ethyl)cyclohexyl)urea (compound b), comprising the step of hydrolyzing a cyclohexane derivative of formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof to obtain trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazin-1-yl)ethyl)cyclohexyl)urea:

[0008]

[0009] in,

[0010] R1, R2 and R3 are each independently selected from H or a leaving group, and at least one of R1, R2 and R3 is a leaving group;

[0011] Optionally, any two of said R1, R2 and R3 may be connected to form a ring;

[0012] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0013]

[0014] The R1 is a leaving group;

[0015] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0016]

[0017] The R3 is a leaving group;

[0018] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0019]

[0020] Said R1 and R2 are each independently selected from a leaving group, and optionally, R1 and R2 can be connected to form a ring;

[0021] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0022]

[0023] Said R1 and R3 are each independently selected from a leaving group, and optionally, R1 and R3 can be connected to form a ring;

[0024] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0025]

[0026] Said R1, R2 and R3 are each independently selected from a leaving group, and optionally, R1 and R2 may be connected to form a ring, and / or R1 and R3 may be connected to form a ring, and / or R2 and R3 may be connected to form a ring;

[0027] The connection to form a ring may be connection to form a five-membered, six-membered, seven-membered or eight-membered heterocyclic ring;

[0028] In some specific embodiments, the leaving group is selected from hydroxyl, thiol, alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy, haloalkylthio, aldehyde, ester, thioester, acyl, ester acyl, amide, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, cycloalkyl, cycloalkyloxy, cycloalkylthio, heterocyclyl, heterocyclyloxy, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy or heteroarylthio, preferably alkyl, alkoxy, haloalkyl, haloalkoxy, aldehyde, ester, acyl, ester acyl, amide, cycloalkyl, cycloalkyloxy, heterocyclyl, heterocyclyloxy, aryloxy, heteroaryloxy; optionally, it can be further substituted;

[0029] In some specific embodiments, the hydroxyl, thiol, alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy, haloalkylthio, aldehyde, ester, thioester, acyl, ester acyl, amide, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, cycloalkyl, cycloalkyloxy, cycloalkylthio, heterocyclyl, heterocyclyloxy, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy or heteroarylthio can be further substituted by one or more of the following groups:

[0030] hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic group, aryl or heteroaryl; optionally, it may be further substituted, for example, further substituted by one or more of hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic group, aryl or heteroaryl;

[0031] In some specific embodiments, the removable group is selected from hydroxyl, thiol, C1-20 alkyl, C1-20 alkoxy, C1-20 alkylthio, C1-20 haloalkyl, C1-20 haloalkoxy, C1-20 haloalkylthio, aldehyde, C1-20 ester, C1-20 thioester, C1-20 acyl, C1-20 ester acyl, C1-20 amide, sulfonic acid, sulfonyl, phosphate, phosphoryl, D-(+)-pyranose glucopyranose group, D-(+)-pyranose glucopyranose ester group, 3-20 membered cycloalkyl, 3-20 membered cycloalkyloxy, 3-20 membered cycloalkylthio, 3-20 membered heterocyclyl, 3-20 membered heterocyclyloxy , 3-20 membered heterocyclylthio, 6-20 membered aryl, 6-20 membered aryloxy, 6-20 membered arylthio, 5-20 membered heteroaryl, 5-20 membered heteroaryloxy or 5-20 membered heteroarylthio, preferably C1-20 alkyl, C1-20 alkoxy, C1-20 haloalkyl, C1-20 haloalkoxy, C1-20 aldehyde, C1-20 ester, C1-20 acyl, C1-20 esteracyl, C1-20 amido, 3-20 membered cycloalkyl, 3-20 membered cycloalkyloxy, 3-20 membered heterocyclyl, 3-20 membered heterocyclyloxy, 6-20 membered aryloxy, 6-20 membered heteroaryloxy; optionally, it may be further substituted;

[0032] In some specific embodiments, the hydroxyl, thiol, C1-20 alkyl, C1-20 alkoxy, C1-20 alkylthio, C1-20 haloalkyl, C1-20 haloalkoxy, C1-20 haloalkylthio, aldehyde, C1-20 ester, C1-20 thioester, C1-20 acyl, C1-20 ester acyl, C1-20 amide, sulfonic acid, sulfonyl, phosphate, phosphoryl, D-(+)-pyranose glucose group , D-(+)-pyranose ester group, 3-20-membered cycloalkyl, 3-20-membered cycloalkyloxy, 3-20-membered cycloalkylthio, 3-20-membered heterocyclyl, 3-20-membered heterocyclyloxy, 3-20-membered heterocyclylthio, 6-20-membered aryl, 6-20-membered aryloxy, 6-20-membered arylthio, 5-20-membered heteroaryl, 5-20-membered heteroaryloxy or 5-20-membered heteroarylthio, etc., may be further substituted with one or more of the following groups:

[0033] hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, D-(+)-pyranose glucopyranose group, D-(+)-pyranose glucopyranose ester group, amino acid group, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, C1-20 acyl, C1-20 acylamino, C1-20 thioester, C1-20 ester, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted, for example further substituted with one or more of hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, D-(+)-pyranose glucopyranose group, D-(+)-pyranose glucopyranose ester group, amino acid group, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, C1-20 acyl, C1-20 acylamino, C1-20 thioester, C1-20 ester, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl;

[0034] In some embodiments, the leaving group can be selected from alkyl or substituted alkyl, alkoxy or substituted alkoxy, aldehyde, acyl or substituted acyl, ester or substituted ester, thioester or substituted thioester, ester acyl or substituted ester acyl, D-(+)-pyranose ester group or substituted D-(+)-pyranose glucopyranose group, D-(+)-pyranose glucopyranose ester group or substituted D-(+)-pyranose glucopyranose ester group, amide or substituted amide, etc.; preferably C1-20 alkyl or substituted C1-20 alkyl, C1-20 Alkoxy or substituted C1-20 alkoxy, aldehyde, C1-20 acyl or substituted C1-20 acyl, C1-20 ester or substituted C1-20 ester, C1-20 thioester or substituted C1-20 thioester, C1-20 ester acyl or substituted C1-20 ester acyl, D-(+)-pyranose ester or substituted D-(+)-pyranose glucopyranose group, D-(+)-pyranose glucopyranose ester or substituted D-(+)-pyranose glucopyranose ester, C1-20 amide or substituted C1-20 amide, etc.;

[0035] The substituents on the substituted alkyl, substituted alkoxy, substituted acyl, substituted ester, substituted thioester, substituted ester acyl, substituted D-(+)-pyranose glucopyranose group, substituted D-(+)-pyranose glucopyranose ester group, substituted amide group, etc. may be one or more of the following groups:

[0036] halogen, hydroxyl, thiol, amino, amine, (C1-20) alkyl, (C1-20) hydroxyalkyl, (C1-20) alkoxy, (C1-20) haloalkyl, (C1-20) ester, (C1-20) thioester, sulfonic acid, sulfonyl, phosphate, phosphoryl, (C1-20) acylamine, D-(+)-pyranose glucopyranose group, D-(+)-pyranose glucopyranose ester group, amino acid group, (3-20 membered) cycloalkyl, (3-20 membered) heterocyclyl, (6-20 membered aryl) or (5-20 membered) heteroaryl, etc.; optionally, it may be further substituted;

[0037] In some embodiments, the leaving group may be

[0038] In some embodiments, the leaving group may be

[0039] In some embodiments, the leaving group can be selected from

[0040] in,

[0041] m is an integer from 0 to 10, specifically 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0042] n is an integer from 1 to 10, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0043] X1 and X2 are each independently selected from -O-, -S- or -NR7-;

[0044] R4, R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it can be further substituted;

[0045] R6 is each independently selected from hydrogen, sulfonyl, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted;

[0046] R7 is each independently selected from hydrogen, hydroxy, thiol, sulfonic acid, sulfonyl, phosphate, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted;

[0047] R8 is each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sugar group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, acyl, acylamino, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted;

[0048] R9 is selected from

[0049] R 10 Selected from -O-,

[0050] Optionally, any two of said R4, R5, R6, R7 and R8 may be connected to form a ring;

[0051] The hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic, aryl or heteroaryl in R4, R5, R6, R7 and R8 may be substituted by one or more of the following groups:

[0052] hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic group, aryl or heteroaryl; optionally, it may be further substituted, for example, further substituted by one or more of hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic group, aryl or heteroaryl;

[0053] In some embodiments, the leaving group may be

[0054] In the present invention, the leaving group may not be a methyl group;

[0055] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0056]

[0057] Wherein, m, R4, R5, and R6 are defined as above;

[0058] Preferably,

[0059] m is 0, 1, 2, 3, 4, 5 or 6;

[0060] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0061] R6 is each independently selected from hydrogen, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0062] More preferably,

[0063] m is 0, 1, 2 or 3;

[0064] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0065] R6 is each independently selected from hydrogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0066] More preferably,

[0067] Each independently selected from

[0068] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0069]

[0070] Wherein, n, R4, R5, and R7 are defined as above;

[0071] Preferably,

[0072] n is 1, 2 or 3;

[0073] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0074] R7 is each independently selected from hydrogen, hydroxyl, mercapto, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0075] More preferably,

[0076] n is 1 or 2;

[0077] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0078] R7 is each independently selected from hydrogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0079] More preferably,

[0080] Each independently selected from

[0081] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0082]

[0083] Wherein, n, R4, R5, R7, and R8 are defined as above;

[0084] Preferably,

[0085] n is 1, 2 or 3;

[0086] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0087] R7 is each independently selected from hydrogen, hydroxyl, mercapto, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0088] R8 is each independently selected from hydrogen, halogen, hydroxyl, mercapto, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0089] More preferably,

[0090] n is 1 or 2;

[0091] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0092] R7 is each independently selected from hydrogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0093] R8 is each independently selected from hydrogen, halogen, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0094] More preferably,

[0095] Each independently selected from

[0096] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0097]

[0098] Wherein, m, R4, R5, and R6 are defined as above;

[0099] Preferably,

[0100] m is 0, 1, 2, 3, 4, 5 or 6;

[0101] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0102] R6 is each independently selected from hydrogen, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0103] More preferably,

[0104] m is 0, 1, 2 or 3, especially 0 or 1;

[0105] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0106] R6 is each independently selected from hydrogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0107] More preferably,

[0108] Each independently selected from

[0109] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0110]

[0111] Wherein, R8 is defined as above;

[0112] Preferably,

[0113] R8 is each independently selected from hydrogen, halogen, hydroxyl, mercapto, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0114] More preferably,

[0115] R8 is each independently selected from hydrogen, halogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0116] More preferably,

[0117] Each independently selected from

[0118] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0119]

[0120] Wherein, m, R4, R5, and R7 are as defined above;

[0121] Preferably,

[0122] m is 0, 1, 2 or 3;

[0123] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0124] R7 is each independently selected from hydrogen, hydroxyl, mercapto, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0125] More preferably,

[0126] m is 0, 1 or 2;

[0127] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0128] R7 is each independently selected from hydrogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0129] More preferably,

[0130] Each independently selected from

[0131] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0132]

[0133] Wherein, m, R4, R5, and R6 are defined as above;

[0134] Preferably,

[0135] m is 0, 1, 2, 3, 4, 5 or 6;

[0136] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0137] R6 is each independently selected from hydrogen, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituents are as described above; or -O-R6 is a sugar group;

[0138] More preferably,

[0139] m is 0, 1, 2 or 3, especially 0 or 1;

[0140] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0141] R6 is each independently selected from hydrogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl; or -O-R6 is a D-(+)-pyranose group;

[0142] The C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclic group, 6-10 membered aryl or 5-10 membered heteroaryl etc. may be substituted by one or more of the following groups:

[0143] hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, D-(+)-pyranose glucopyranose, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0144] More preferably,

[0145] Each independently selected from

[0146] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0147]

[0148]

[0149] Wherein, n, X1, X2, R4, R5, and R8 are as defined above;

[0150] Preferably,

[0151] X1 is -O-, X2 is -O-;

[0152] or,

[0153] X1 is -S-, X2 is -S-;

[0154] or,

[0155] X1 is -O-, X2 is -S-;

[0156] or,

[0157] X1 is -S-, X2 is -O-;

[0158] n is 1, 2 or 3;

[0159] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0160] R8 is each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, amino acid group, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 acylamino, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0161] More preferably,

[0162] n is 1 or 2;

[0163] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0164] R8 is each independently selected from hydrogen, halogen, amino, amine, amino acid group, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl, optionally, it can be further substituted, for example, further by hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, D-(+)-pyranose group, D-(+)-pyranose glucopyranose ester group, amino acid group, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, C1-10 acyl, C1-10 acylamino, C1-10 thioester, C1-10 ester, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl are substituted by one or more of the following groups;

[0165] The amino acid group is preferably a phenylalanine group, an alanine group, or a glycine group.

[0166] The 3-10 membered cycloalkyl group may specifically be cyclopropyl or cyclohexyl;

[0167] R8 is preferably independently

[0168] More preferably,

[0169] Each independently selected from

[0170]

[0171] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0172]

[0173] Wherein, R4, R5, and R8 are defined as above;

[0174] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0175]

[0176] Wherein, n, X1, R4, R5, and R8 are defined as above;

[0177] Preferably,

[0178] X1 is -O-;

[0179] n is 1, 2 or 3;

[0180] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0181] R8 is each independently selected from hydrogen, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0182] More preferably,

[0183] n is 1 or 2;

[0184] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0185] R8 is each independently selected from C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl, which may be further substituted, for example, by hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, D-(+)-pyranose glucose group, D-(+)-pyranose glucose group, substituted with one or more of a glucose ester group, an amino acid group, a C1-10 alkyl group, a C1-10 haloalkyl group, a C1-10 hydroxyalkyl group, a C1-10 alkoxy group, a C1-10 hydroxyalkoxy group, a C1-10 haloalkoxy group, a C1-10 acyl group, a C1-10 acylamino group, a C1-10 thioester group, a C1-10 ester group, a 3-10 membered cycloalkyl group, a 3-10 membered heterocyclyl group, a 6-10 membered aryl group or a 5-10 membered heteroaryl group;

[0186] More preferably,

[0187] for

[0188] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0189]

[0190]

[0191] Wherein, n, X1, R4, R5, and R6 are defined as above;

[0192] Preferably,

[0193] X1 is -O-;

[0194] n is 1, 2 or 3;

[0195] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0196] R6 is each independently selected from hydrogen, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, it may be further substituted; preferably, the further substituted substituent is as described above;

[0197] More preferably,

[0198] n is 1 or 2;

[0199] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0200] R6 is each independently selected from hydrogen, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0201] The C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, 3-10 membered cycloalkyl, 3-10 membered heterocyclic group, 6-10 membered aryl or 5-10 membered heteroaryl etc. may be substituted by one or more of the following groups:

[0202] hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, C1-10 ester, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0203] More preferably,

[0204] Each independently selected from

[0205] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0206]

[0207] in,

[0208] R 11 is a removable linking group;

[0209] g is selected from 2, 3 or 4;

[0210] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0211]

[0212] in,

[0213] R9 is selected from

[0214] R 10 Selected from -O-,

[0215] n, X1, X2, R4, and R5 are as defined above;

[0216] Preferably,

[0217] X1 and X2 are -O-;

[0218] n is 1, 2, 3, 4 or 5;

[0219] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-20 alkyl, C1-20 haloalkyl, C1-20 hydroxyalkyl, C1-20 alkoxy, C1-20 hydroxyalkoxy, C1-20 haloalkoxy, 3-20 membered cycloalkyl, 3-20 membered heterocyclyl, 6-20 membered aryl or 5-20 membered heteroaryl; optionally, they may be further substituted; preferably, the further substituted substituents are as described above;

[0220] More preferably,

[0221] n is 1, 2, 3 or 4;

[0222] R4 and R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, mercapto, cyano, amino, C1-10 alkyl, C1-10 haloalkyl, C1-10 hydroxyalkyl, C1-10 alkoxy, C1-10 hydroxyalkoxy, C1-10 haloalkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl;

[0223] In the present invention, or R 10 Each can be independently selected from

[0224] In the present invention, any two of R4, R5, R6, R7 and R8 can be connected to form a ring; preferably a five-membered, six-membered, seven-membered or eight-membered ring;

[0225] In some specific embodiments, the structure of the cyclohexane derivative is as follows:

[0226]

[0227]

[0228]

[0229]

[0230] In the present invention, the cyclohexane derivatives may not include the following compounds:

[0231]

[0232] In the present invention, the function of the hydrolysis is to remove the leaving groups; for example, to remove the leaving groups R1, R2 and / or R3 in the above-mentioned cyclohexane derivatives;

[0233] In the present invention, the hydrolysis may be acid hydrolysis or alkaline hydrolysis; the acid hydrolysis is to hydrolyze and remove the leaving group under acid conditions; the alkaline hydrolysis is to hydrolyze and remove the leaving group under alkaline conditions;

[0234] In the acid hydrolysis, the acid may be an inorganic acid or an organic acid;

[0235] The inorganic acid is preferably hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid or phosphoric acid, more preferably hydrochloric acid or sulfuric acid, and further preferably hydrochloric acid;

[0236] The organic acid may be oxalic acid, maleic acid, acetic acid, formic acid or tartaric acid;

[0237] In the acid hydrolysis, the base is preferably an inorganic base, and the inorganic base is preferably sodium hydroxide, potassium hydroxide, or sodium hydrogen;

[0238] The molar ratio of the acid / base to the cyclohexane derivative may be 1:100 to 100:1, preferably 10-20:1;

[0239] When acid hydrolysis is used, after hydrolysis, optionally, the pH of the system is adjusted to 7-10, preferably 8, with a base; wherein the base for adjusting the pH can be sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, etc.;

[0240] The hydrolysis can be carried out in a solvent, and the solvent can be water, an organic solvent or a mixture thereof; the organic solvent can be methanol, ethanol, tetrahydrofuran, ethyl acetate, acetonitrile, etc.; the solvent is preferably methanol, a mixture of tetrahydrofuran and methanol, or a mixture of methanol and water; the volume ratio of tetrahydrofuran to methanol in the mixture of tetrahydrofuran and methanol can be 1:10 to 10:1, preferably 1:3 to 3:1; the volume ratio of methanol to water in the mixture of methanol and water can be 1:10 to 10:1, preferably 1:3 to 3:1;

[0241] The reaction temperature of the hydrolysis is -20°C to 100°C, preferably 0°C to 50°C, more preferably 10°C to 30°C;

[0242] The reaction time of the hydrolysis is 0.1-10 hours, preferably 0.5-2 hours.

[0243] Detailed description of the invention

[0244] Before further describing the present invention, it should be understood that the present invention is not limited to the specific embodiments described, as they may of course vary. It should also be understood that the purpose of the terms used herein is only to describe specific embodiments and is not intended to be limiting, as the scope of the present invention is limited only by the appended claims.

[0245] Unless otherwise indicated, all scientific and technological terms used herein have the same ordinary meaning as understood by those of ordinary skill in the art to which the present invention belongs. All patents, applications, published applications and other publications cited herein are incorporated herein by reference in their entirety. If the definitions in this section are contrary to or inconsistent with the definitions listed in the patents, applications and other publications incorporated herein by reference, the definitions in this section will prevail over the definitions incorporated herein by reference.

[0246] In the present invention, "alkyl" refers to a saturated aliphatic hydrocarbon group, which may be a C1-20 alkyl group, preferably a C1-10 alkyl group, more preferably a C1-6 alkyl group, and most preferably a C1-3 alkyl group. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof;

[0247] In the present invention, "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent. The cycloalkyl group may contain 3 to 20 ring carbon atoms ("3-20-membered cycloalkyl"), preferably 3 to 12 ring carbon atoms ("3-12-membered cycloalkyl"), further preferably 3 to 10 ring carbon atoms ("3-10-membered cycloalkyl"), more preferably 3 to 8 ring carbon atoms ("3-8-membered cycloalkyl"), and most preferably 3 to 6 ring carbon atoms ("3-6-membered cycloalkyl"). Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.;

[0248] In the present invention, "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing a specified number of ring atoms and including at least one heteroatom selected from N, O and S as a ring member of the cycloalkyl ring. The heterocyclyl may contain 3 to 20 ring atoms ("3-20 membered heterocyclyl"), preferably 3 to 12 ring atoms ("3-12 membered heterocyclyl"), further preferably 3 to 10 ring atoms ("3-10 membered heterocyclyl"), more preferably 3 to 8 ring atoms ("3-8 membered heterocyclyl"), and most preferably 3 to 6 ring atoms ("3-6 membered heterocyclyl"). Non-limiting examples of heterocyclyl include oxirane, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, etc.;

[0249] In the present invention, "aryl" refers to an optionally substituted monocyclic, biaryl or fused bicyclic or polycyclic ring system having the well-known aromatic characteristics, wherein at least one ring contains a completely conjugated π-electron system. Typically, aryl contains 6 to 20 carbon atoms ("6-20 membered aryl") as ring members, preferably 6 to 14 carbon atoms ("6-C membered aryl") or more preferably 6 to 12 carbon atoms ("6-10 membered aryl"). Fused aryl may include an aryl ring fused to another aryl ring, or to a saturated or partially unsaturated carbocyclic or heterocyclic ring. The point of attachment to the base molecule on such a fused aryl ring system may be a C atom of the aromatic portion of the ring system or a C or N atom of the non-aromatic portion. Examples of aryl include, but are not limited to, phenyl, biphenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl, indenyl, benzo[d][1,3]dioxole and tetrahydronaphthyl;

[0250] In the present invention, " heteroaryl " refers to monocyclic, heterobiaryl or fused bicyclic or polycyclic ring system with well-known aromaticity characteristics, which contains a specified number of ring atoms and includes at least one heteroatom selected from N, O and S as a ring member in the aromatic ring. The inclusion of heteroatoms allows the aromaticity of 5-ring and 6-ring. Usually, heteroaryl contains 5-20 ring atoms (" 5-20 yuan heteroaryl "), preferably 5-14 ring atoms (" 5-14 yuan heteroaryl "), and more preferably 5-10 ring atoms (" 5-10 yuan heteroaryl "). The heteroaryl ring is connected to the base molecule by the ring atoms of the heteroaromatic ring, thereby keeping aromaticity. Therefore, 6 yuan heteroaryl rings can be connected to the base molecule by ring C atoms, and 5 yuan heteroaryl rings can be connected to the base molecule by ring C or N atoms. Examples of unsubstituted heteroaryl groups often include, but are not limited to, pyrrole, furan, thiophene, pyrazole, imidazole, isoxazole, oxazole, isothiazole, thiazole, triazole, oxadiazole, thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, benzofuran, benzothiophene, indole, benzimidazole, indazole, quinoline, isoquinoline, purine, triazine, naphthyridine, and carbazole;

[0251] In the present invention, "ester group" refers to a group formed by esterification of a carboxyl group (-COOH) and a hydroxyl group (-OH), with the OH group removed and the H group removed. The ester group can be wherein R6 and R8 are as defined above, and may be selected from a C1-20 ester group, preferably a C1-10 ester group, further preferably a C2-8 ester group, more preferably a C2-6 ester group, and most preferably a C2-3 ester group;

[0252] In the present invention, the "thioester group" can be wherein R6 and R8 are as defined above, and may be selected from C1-20 thioester groups, preferably C1-10 thioester groups, further preferably C2-8 thioester groups, more preferably C2-6 thioester groups, and most preferably C2-3 thioester groups;

[0253] In the present invention, "acyl" refers to the atomic group remaining after removing the hydroxyl group from an organic acid, and has the general formula R8-CO-, wherein R8 is defined as described above, and the acyl group can be selected from C1-20 acyl, preferably C1-10 acyl, further preferably C1-8 acyl, more preferably C1-6 acyl, and most preferably C1-3 acyl;

[0254] In the present invention, "aldehyde group" refers to -C(O)H;

[0255] In the present invention, "amino acid group" refers to a compound formed when the hydrogen atom on the carbon atom of a carboxylic acid is replaced by an amino group, and "amino acid group" refers to a group formed by removing a hydrogen atom on the carbon atom in an amino acid, and amino acids include glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine. Correspondingly, the groups obtained based on amino acids include glycine group, alanine group, valine group, leucine group, isoleucine group, methionine group, proline group, tryptophan group, serine group, tyrosine group, cysteine ​​group, phenylalanine group, asparagine group, glutamine group, threonine group, aspartic acid group, glutamine group, threonine group, aspartic acid group, glutamic acid group, lysine group, arginine group, histidine group, selenocysteine ​​group, pyrrolysine group, etc., and specifically can be

[0256] In the present invention, "sugar group" refers to a group formed by removing one hydrogen from the hydroxyl group of a sugar molecule, and can be a glucose group, a fructose group, a galactose group, a sucrose group, a maltose group, etc., and can be specifically

[0257] In the present invention, "sugar ester group" refers to a -CO-sugar group, wherein the sugar group is as defined above;

[0258] In the present invention, "alkoxy" refers to alkyl-O-, wherein alkyl is as defined above; cycloalkyloxy, heterocyclyloxy, aryloxy, heteroaryloxy are similarly defined;

[0259] In the present invention, "alkylthio" refers to alkyl-S-, wherein the alkyl group is as defined above; cycloalkylthio, heterocyclylthio, arylthio, and heteroarylthio are similarly defined;

[0260] In the present invention, "amide group" refers to wherein R7 is as defined above;

[0261] In the present invention, "acylamino group" refers to wherein R7 and R8 are as defined above;

[0262] In the present invention, "amino group" refers to wherein R7 is as defined above, and at least one R7 is not H;

[0263] In the present invention, "haloalkyl" refers to an alkyl group substituted by one or more halogens, wherein alkyl is as defined above;

[0264] In the present invention, "haloalkoxy" refers to an alkoxy group substituted by one or more halogens, wherein the alkoxy group is as defined above;

[0265] In the present invention, "haloalkylthio" refers to an alkylthio group substituted by one or more halogens, wherein the alkylthio group is as defined above;

[0266] As used herein, "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups, wherein the alkyl group is as defined above;

[0267] As used herein, "hydroxyalkoxy" refers to an alkoxy group substituted with one or more hydroxy groups, wherein the alkoxy group is as defined above;

[0268] In the present invention, "ester acyl" refers to an ester group -CO-, wherein the ester group is as defined above;

[0269] In the present invention, "hydroxyl" refers to -OH;

[0270] In the present invention, "thiol" refers to -SH;

[0271] In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine;

[0272] In the present invention, "amino" refers to -NH2;

[0273] In the present invention, "cyano" refers to -CN;

[0274] In the present invention, "nitro" refers to -NO2;

[0275] In the present invention, "sulfonic acid group" refers to -SO3H;

[0276] In the present invention, "sulfonyl" refers to R8-S(=O)2-, wherein R8 is as defined above, and the sulfonyl group can specifically be methylsulfonyl, phenylsulfonyl, p-toluenesulfonyl, etc.;

[0277] In the present invention, "phosphate group" refers to wherein R6 and R8 are as defined above;

[0278] In the present invention, "phosphoryl" refers to wherein R6 and R8 are as defined above;

[0279] In the present invention, the above-defined groups may be optionally substituted or unsubstituted, and when substituted, may be substituted by one or more of the following groups:

[0280] Hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic group, aryl or heteroaryl; optionally, it can be further substituted.

[0281] In the present invention, any isotope-labeled derivatives of the compounds of the present invention or their pharmaceutically acceptable salts are covered by the present invention. Atoms that can be isotopically labeled include, but are not limited to, hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, iodine, etc. They can be labeled with isotopes. 2 H(D), 3 H. 11 C. 13 C. 14 C. 15 N. 18 F. 31 P. 32 P. 35 S. 36 Cl and 125 Unless otherwise indicated, when a position is specifically designated as deuterium (D), the position is understood to have an abundance of deuterium (i.e., at least 45% deuterium incorporation) that is at least 3000 times greater than the natural abundance of deuterium, which is 0.015%.

[0282] In the present invention, "plurality" means two or more, for example, 2, 3, 4, 5 or 6, etc.

[0283] In the present invention, "optional" or "optionally" means that the event or circumstance described later may but need not occur, and the description includes occasions where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes situations where the heterocyclic group is substituted with an alkyl group and situations where the heterocyclic group is not substituted with an alkyl group.

[0284] In the present invention, "pharmaceutically acceptable salts" refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.

[0285] In the present invention, the following abbreviations / terms are used:

[0286] acetyl chloride: acetyl chloride

[0287] pyridine: pyridine

[0288] Sulfuryl chloride: Sulfuryl chloride

[0289] Triphosgene: triphosgene

[0290] Acetone: Acetone

[0291] THF: Tetrahydrofuran

[0292] DCM: dichloromethane

[0293] ACN: Acetonitrile

[0294] DMF: N,N-dimethylformamide

[0295] TEA: triethylamine

[0296] DIEA: N,N-diisopropylethylamine

[0297] MTBE: Methyl tert-butyl ether DETAILED DESCRIPTION

[0298] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples.

[0299] The experimental methods in the examples of the present invention without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by raw material or commodity manufacturers. Reagents without specifying specific sources are conventional reagents purchased from the market.

[0300] Example 1

[0301] Preparation of trans-4-(2-(4-(Benzo[b]thiophen-7-yl)piperazin-1-yl)ethyl)cyclohexyl)urea (Intermediate 1, also referred to as Compound b)

[0302]

[0303] 9.3 g of trans-4-[2-[4-(Benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexylamine (Compound a) was dissolved in a mixed solvent of DMF:water = 3:1 (37.2 mL), 2.71 g of potassium cyanate was added, and the mixture was heated at 85°C for 2 h, and 15 g of potassium cyanate was added to react at 85°C overnight. The mixture was stopped from heating, cooled to room temperature, stirred with water, filtered, the filter cake was rinsed with water, and dried to obtain 6.5 g of Intermediate 1.

[0304] MS (ESI): m / z = 387.3 [M+H] + .

[0305] H NMR (400MHz, d-TFA) δ8.33(d,J=7.6Hz,1H),7.91-7.79(m,4H),5.08(t,J=12Hz,2H),4.68-4.56(m,4H),4.40(t ,J=12Hz,2H),3.84-3.82(m,3H),2.41(d,J=10Hz,2H),2.19-2.12(m,4H),1.70-1.64(m,3H),1.52-1.46(m,2H).

[0306] 13 C-NMR(101MHz,d-TFA)δ159.675,144.647,132.903,129.461,127.624,126.844,125.74 8,125.389,117.161,56.978,51.445,50.941,49.697,33.981,31.246,30.353,30.154.

[0307] Example 2

[0308]

[0309] In a 50mL three-necked flask, intermediate 1 (0.200g, 0.517mmol) was dissolved in tetrahydrofuran (5mL), pyridine (0.287g, 3.622mmol) was added, acetyl chloride (0.256g, 3.257mmol) was added under ice bath, and the mixture was reacted at room temperature for 1h. After the reaction was completed, the solvent was removed by rotary evaporation, 20mL of water was added, and the mixture was stirred at room temperature for 1h to precipitate a white solid. After filtering, the filter cake was washed with water and dried to obtain compound 1 (88.98mg, yield: 40.2%).

[0310] LC-MS:429.2[M+H] + .

[0311] 1 H NMR(400MHz,TFA-d1)δ8.17(d,J=8.0Hz,1H),7.89(d,J=7.8Hz,1H),7.78–7.59(m,3H),5.02(s,2H),4.43(d,J=40.7Hz,6H), 3.85(s,1H),3.66(s,2H),2.41(s,3H),2.32-2.17(m,2H),2.01(d,J=15.2Hz,4H),1.64-1.45(m,3H),1.34(t,J=12.6Hz,2H).

[0312] Example 3

[0313]

[0314] In a 50mL three-necked flask, intermediate 1 (0.200g, 0.517mmol) was dissolved in dichloromethane (5mL), pyridine (1.475g, 18.647mmol) was added, chloromethyl chloroformate (0.75g, 7.937mmol) was added under ice bath, and refluxed for 30min. After the reaction was completed, the solvent was removed by rotary evaporation, 20mL of water was added, and the mixture was stirred at room temperature for 1h to precipitate a white solid. After filtering, the filter cake was washed with water and dried to obtain compound 2 (206.50mg, yield: 89.84%).

[0315] LC-MS:445.3[M+H] + .

[0316] 1 H NMR(400MHz,TFA-d1)δ8.20(d,J=8.0Hz,1H),7.92(d,J=7.6Hz,1H),7.81-7.56(m,3H),5.05(s,2H),4.46(d,J=39.8Hz,6H),4.01(s, 3H), 3.89 (s, 1H), 3.69 (s, 2H), 2.25 (d, J = 12.1Hz, 2H), 2.04 (d, J = 15.4Hz, 4H), 1.55 (dd, J = 24.8, 12.7Hz, 3H), 1.35 (d, J = 14.3Hz, 2H).

[0317] Example 4

[0318]

[0319] Step 1: Preparation of intermediate 2

[0320] In a 100mL three-necked flask, intermediate 1 (0.500g, 1.293mmol) was dissolved in tetrahydrofuran (15mL), and pyridine (1.966g, 24.855mmol) was added; chloroacetyl chloride (1.419g, 12.564mmol) was added under ice bath, and the reaction was carried out at room temperature for 1h. After the reaction was completed, water was added to dilute and extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was obtained by column chromatography to obtain intermediate 2 (521.00mg, yield: 87.02%).

[0321] LC-MS:463.3[M+H] + .

[0322] Step 2: Preparation of compound 3

[0323] In a 50mL three-necked flask, intermediate 2 (0.100g, 0.216mmol) was dissolved in tetrahydrofuran (5mL), and 3mL of ammonia water was added; the reaction was carried out at 50°C for 2h. After the reaction was completed, water was added to dilute and extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and dried. The crude product was purified by preparative liquid phase to obtain compound 3 (23.46mg, yield: 24.49%).

[0324] LC-MS:444.3[M+H] + .

[0325] 1 H NMR (400 MHz, TFA-d1) δ 8.13 (d, J = 7.9 Hz, 1H), 7.77-7.54 (m, 4H), 4.89 (t, J = 12.8 Hz, 2H), 4.55-4.12 (m, 8H), 3.81 (s, 1H), 3.68-3.54 (m, 2H), 2.19 (d, J = 12.2 Hz, 2H), 2.03-1.85 (m, 4H), 1.62-1.37 (m, 3H), 1.28 (q, J = 12.5 Hz, 2H). Example 5

[0326]

[0327] Step 1: Preparation of intermediate 2

[0328] In a 100mL three-necked flask, intermediate 1 (0.500g, 1.293mmol) was dissolved in tetrahydrofuran (15mL), and pyridine (1.966g, 24.855mmol) was added; chloroacetyl chloride (1.419g, 12.564mmol) was added under ice bath, and the reaction was carried out at room temperature for 1h. After the reaction was completed, water was added to dilute and extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was obtained by column chromatography to obtain intermediate 2 (521.00mg, yield: 87.02%).

[0329] LC-MS:463.3[M+H] + .

[0330] Step 2: Preparation of compound 4

[0331] In a 50mL three-necked flask, intermediate 2 (0.100g, 0.216mmol) was dissolved in acetonitrile (5mL), potassium carbonate (0.239g, 1.728mmol), methylamine hydrochloride (0.073g, 1.080mmol) and potassium iodide (0.004g, 0.022mmol) were added; the reaction was carried out at 50°C for 2h. After the reaction was completed, water was added to dilute and extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by preparative liquid phase to obtain compound 4 (20.64mg, yield: 20.88%).

[0332] LC-MS:458.3[M+H] + .

[0333] 1 H NMR(400MHz,TFA-d1)δ8.13(d,J=7.9Hz,1H),7.79-7.52(m,4H),4.99-4.83(m,2H),4.52-4.07(m,8H),3.80(s, 1H),3.67-3.54(m,2H),3.12(s,3H),2.24-2.09(m,2H),2.04-1.84(m,4H),1.35(dq,J=65.0,12.6,11.8Hz,5H).

[0334] Example 6

[0335]

[0336] In a 50mL three-necked flask, intermediate 1 (0.100g, 0.259mmol) was dissolved in tetrahydrofuran (6mL), and pyridine (0.492g, 6.220mmol) was added; propionyl chloride (0.533g, 5.761mmol) was added under ice bath, and the reaction was carried out at room temperature for 1h. After the reaction was completed, the solvent was removed by rotary evaporation, 20mL of water was added, and the mixture was stirred at room temperature for 1h to precipitate a white solid. After filtering, the filter cake was washed with water, and the wet filter cake was slurried with 15mL of methanol at room temperature for 1h, filtered and dried to obtain compound 5 (62.00mg, yield: 54.08%).

[0337] LC-MS:443.3[M+H] + .

[0338] 1H NMR(400MHz,TFA-d1)δ8.23–8.12(m,1H),7.90(d,J=7.8Hz,1H),7.77–7.51(m,3H),5.03(s,2H),4.55–4.27(m,6H),3.85(s,1H),3.6 6(s,2H),2.66(q,J=7.6Hz,2H),2.23(d,J=12.3Hz,2H),2.01(d,J=14.7Hz,4H),1.52(q,J=12.5,11.8Hz,3H),1.35(t,J=7.5Hz,3H).

[0339] Example 7

[0340]

[0341] In a 50mL three-necked flask, intermediate 1 (0.100g, 0.259mmol) was dissolved in tetrahydrofuran (5mL), and pyridine (0.492g, 6.220mmol) was added; n-butyryl chloride (0.513g, 4.815mmol) was added under ice bath, and the temperature was raised to 50°C for reaction for 1h. After the reaction was completed, the solvent was removed by rotary evaporation, 20mL of water was added, and the mixture was stirred at room temperature for 1h to precipitate a white solid. After filtering, the filter cake was washed with water, and the wet filter cake was slurried with 15mL of methanol at room temperature for 1h, filtered and dried to obtain compound 6 (52.61mg, yield: 44.48%).

[0342] LC-MS:457.3[M+H] + .

[0343] 1 H NMR(400MHz,TFA-d1)δ8.16(d,J=8.0Hz,1H),7.88(d,J=7.9Hz,1H),7.72(d,J=5.5Hz,1H) ,7.69–7.59(m,2H),5.05–4.99(m,2H),4.46(d,J=13.6Hz,2H),4.37(d,J=7.0Hz,4H),3.90 –3.80(m,1H),3.65(t,J=8.3Hz,2H),2.58(dt,J=12.5,7.5Hz,2H),2.23(d,J=12.3Hz,2H), 2.08–1.76(m,7H),1.51(q,J=12.9,12.1Hz,2H),1.38–1.25(m,2H),1.11(t,J=7.4Hz,3H).

[0344] Example 8

[0345]

[0346] In a 50mL three-necked flask, intermediate 1 (0.100g, 0.259mmol) was dissolved in tetrahydrofuran (6mL), and pyridine (0.492g, 6.220mmol) was added; isobutyryl chloride (0.509g, 4.777mmol) was added under ice bath, and the temperature was raised to 50°C for reaction for 1h. After the reaction was completed, the solvent was removed by rotary evaporation, 20mL of water was added, and the mixture was stirred at room temperature for 1h to precipitate a white solid. After filtering, the filter cake was washed with water, and the wet filter cake was slurried with 15mL of methanol at room temperature for 1h, filtered and dried to obtain compound 7 (45.37mg, yield: 37.06%).

[0347] LC-MS:457.4[M+H] + .

[0348] 1 H NMR(400MHz,TFA-d1)δ8.16(d,J=8.0Hz,1H),7.68(dt,J=40.8,7.1Hz,4H),4.92(t,J=13.3Hz,2H),4.61–4.16(m,6H),3.83( s,1H),3.64(d,J=9.0Hz,2H),2.95–2.70(m,1H),2.22(d,J=12.5Hz,2H),1.96(dd,J=22.4,11.7Hz,4H),1.68–1.17(m,11H).

[0349] Example 9

[0350]

[0351] In a 50mL three-necked flask, intermediate 1 (0.100g, 0.259mmol) was dissolved in tetrahydrofuran (5mL), and pyridine (0.492g, 6.220mmol) was added; propyl chloroformate (0.545g, 4.447mmol) was added under ice bath, and the temperature was raised to 50°C for reaction for 1h. After the reaction was completed, the solvent was removed by rotary evaporation, 20mL of water was added, and the mixture was stirred at room temperature for 1h to precipitate a white solid. After filtering, the filter cake was washed with water, and the wet filter cake was slurried with 15mL of methanol at room temperature for 1h, filtered and dried to obtain compound 8 (50.32mg, yield: 41.11%).

[0352] LC-MS:473.3[M+H] + .

[0353] 1H NMR(400MHz,TFA-d1)δ8.16(d,J=8.1Hz,1H),7.79–7.55(m,4H),4.92(t,J =12.9Hz,2H),4.54–4.45(m,2H),4.40(d,J=13.5Hz,2H),4.35–4.19(m,4H ),3.93–3.78(m,1H),3.68–3.59(m,2H),2.21(d,J=12.5Hz,2H),2.03–1.7 7(m,7H),1.63–1.43(m,4H),1.30(q,J=12.5Hz,2H),1.08(d,J=7.4Hz,3H).

[0354] Example 10

[0355]

[0356] Step 1: Preparation of intermediate 2

[0357] In a 100mL three-necked flask, intermediate 1 (0.500g, 1.293mmol) was dissolved in tetrahydrofuran (15mL), and pyridine (1.966g, 24.855mmol) was added; chloroacetyl chloride (1.419g, 12.564mmol) was added under ice bath, and the reaction was carried out at room temperature for 1h. After the reaction was completed, water was added to dilute and extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was obtained by column chromatography to obtain intermediate 2 (521.00mg, yield: 87.02%).

[0358] LC-MS:463.3[M+H] + .

[0359] Step 2: Preparation of compound 9

[0360] In a 50mL three-necked flask, intermediate 1 (0.100g, 0.259mmol) was dissolved in acetonitrile (5mL), potassium carbonate (0.358g, 2.592mmol), dimethylamine hydrochloride (0.169g, 2.073mmol) and potassium iodide (0.004g, 0.026mmol) were added; the mixture was reacted at 50°C for 2h. After the reaction was completed, the solvent was removed by rotary evaporation, 20mL of water was added, and the mixture was stirred at room temperature for 1h to precipitate a gray-brown solid. After filtering, the filter cake was washed with water and dried to obtain compound 9 (86.30mg, yield: 70.65%).

[0361] LC-MS:472.3[M+H] + .

[0362] 1H NMR(400MHz,TFA-d1)δ8.13(d,J=8.0Hz,1H),7.84–7.50(m,4H),4.89(t,J=13.0Hz,2H),4.56–4.29(m,6H),4.30–4.00(m, 2H),3.81(s,1H),3.61(dd,J=10.1,6.2Hz,2H),3.24(s,6H),2.19(d,J=12.5Hz,2H),2.01–1.86(m,5H),1.62–1.19(m,4H).

[0363] Embodiment 11

[0364]

[0365] Step 1: Preparation of intermediate 5

[0366] In a 500mL three-necked flask, intermediate 3 (10.022g, 161.3mmol) was dissolved in methyl tert-butyl ether (200mL), and DIPEA (22.9g, 177.4mmol) was added; intermediate 4 (21.842g, 169.4mmol) was added under ice bath, and the reaction was carried out at room temperature for 16h. After the reaction was completed, 1N HCl (50mL) was added to quench the reaction, the reaction solution was extracted with methyl tert-butyl ether, and the organic phase was washed with 1N HCl (50mL) and saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and the organic phase was spin-dried to obtain intermediate 5 (25.8g, yield: 99.5%).

[0367] 1 H NMR (400MHz, CDCl3) δ5.74 (s, 2H), 2.90 (q, J = 4.0, 12.0Hz, 2H), 1.30 (t, J = 8.0Hz, 3H).

[0368] Step 2: Preparation of intermediate 6

[0369] In a 500mL single-mouth bottle, intermediate 5 (25.990g, 168.1mmol) was dissolved in acetone (200mL), sodium iodide (50.393g, 336.2mmol) was added, and the reaction was carried out at 40°C for 1.5h. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed twice with acetone; after the filtrate was spin-dried, it was dissolved in methyl tert-butyl ether and washed with water. The organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried to obtain intermediate 6 (36g, yield: 87.03%).

[0370] 1H NMR (400MHz, CDCl3) δ5.97 (s, 2H), 2.89 (q, J = 4.0, 12.0Hz, 2H), 1.34 (t, J = 8.0Hz, 3H).

[0371] Step 3: Preparation of intermediate 7

[0372] In a 100mL single-mouth bottle, intermediate 6 (7.997g, 32.5mmol) was dissolved in DMF (40mL), and sodium acetate (3.199g, 39.0mmol) was added. The reaction was carried out at room temperature for 16h. After the reaction was completed, water was added to quench the reaction. The reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was spin-dried. The crude product was obtained by column chromatography to obtain intermediate 7 (3.2g, yield: 55.25%).

[0373] 1 H NMR (400MHz, CDCl3) δ5.77(s,2H),2.86(q,J=4.0,12.0Hz,2H),2.10(s,3H),1.29(t,J=8.0Hz,3H).

[0374] Step 4: Preparation of intermediate 8

[0375] In a 100 mL single-necked bottle, intermediate 7 (3.190 g, 17.9 mmol) was dissolved in dichloromethane (20 mL), and sulfonyl chloride (3.617 g, 26.8 mmol) was added under ice bath. The reaction was carried out at room temperature for 3 h. After the reaction was completed, the solvent was dried to obtain intermediate 8 (2.2 g, yield: 80.58%).

[0376] 1 H NMR (400MHz, CDCl3) δ5.78 (s, 2H), 2.16 (s, 3H).

[0377] Step 5: Preparation of compound 10

[0378] In a 100mL single-mouth bottle, intermediate 1 (0.500g, 1.293mmol) was dissolved in dichloromethane (10mL), pyridine (0.306g, 3.87mmol) was added, and intermediate 8 (2.196g, 14.4mmol) was added under ice bath, and the reaction was carried out at room temperature for 10min. After the reaction was completed, water was added to quench the reaction, and dichloromethane was extracted. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by column chromatography to obtain compound 10 (107.4mg, yield: 16.53%).

[0379] LC-MS: 503.2[M+H] + .

[0380] 1 H NMR (400MHz, DMSO-d6) δ10.28(s,1H),7.71(d,J=8.0Hz,1H),7.55(d,J=4.0Hz,1H), 7.46-7.43(m,2H),7.34-7.31(m,1H),6.97(d,J=8.0Hz,1H),5.68(s,2H),3.49-3.4 3(m,1H),3.14(s,4H),2.58(s,4H),2.39(s,2H),2.08(s,3H),1.87(d,J=12.0Hz,2H ),1.77(d,J=12.0Hz,2H),1.40-1.36(m,2H),1.23-1.16(m,3H),1.05-0.96(m,2H).

[0381] Example 12

[0382]

[0383] Intermediate 9 (0.496g, 5.5mmol) was dissolved in dichloromethane (10mL), pyridine (0.649g, 8.2mmol) was added, triphosgene (0.801g, 2.7mmol) was added at 0°C, and the reaction was carried out at room temperature for 1h. After the reaction was completed, the solid was filtered off and the filtrate was collected and dried. Intermediate 1 (0.100g, 0.259mmol) was dissolved in dichloromethane (10mL), pyridine (0.098g, 1.239) was added, the product of the previous step was added at 0°C, and the reaction was carried out at room temperature for 1h. The reaction solution was diluted with water and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered and dried. The crude product was subjected to column chromatography to obtain compound 11 (27.6mg, yield: 21.12%).

[0384] LC-MS: 503.3 [M+H] + .

[0385] 1H NMR (400MHz, DMSO-d6) δ9.92 (s, 1H), 7.71 (d, J = 4.0Hz, 1H), 7.60-7.54 (m, 2H), 7.44 (d, J=4.0Hz,1H),7.35-7.31(m,1H),6.97(d,J=8.0Hz,1H),4.93-4.89(m,1H),3.46-3.42(m ,1H),3.38-3.35(m,2H),3.26(s,3H),3.14(s,4H),2.59(s,4H),2.40(s,2H),1.88-1.74 (m,4H),1.42-1.36(m,2H),1.29-1.23(m,3H),1.16(d,J=8.0Hz,3H),1.05-0.99(m,2H).

[0386] Embodiment 13

[0387]

[0388] In a 50mL three-necked flask, intermediate 1 (0.100g, 0.259mmol) was dissolved in tetrahydrofuran (5mL), and pyridine (0.492g, 6.220mmol) was added; ethyl chloroformyl acetate (0.585g, 3.885mmol) was added under ice bath, and the reaction was carried out at room temperature for 1h. After the reaction was completed, water was added to dilute and extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by preparative liquid phase to obtain compound 12 (20.00mg, yield: 15.42%).

[0389] LC-MS: 501.3 [M+H] + .

[0390] 1 H NMR (400MHz, DMSO) δ10.48(s,1H),9.51(s,1H),7.78(d,J=5.4Hz,1H),7.64(d,J= 7.8Hz,1H),7.49(d,J=5.4Hz,1H),7.38(t,J=7.7Hz,1H),7.06(d,J=7.6Hz,1H),4 .10(q,J=7.1Hz,2H),3.68(d,J=12.8Hz,4H),3.45(s,3H),3.12(d,J=12.3Hz,2H) ,1.91(d,J=12.0Hz,2H),1.78(d,J=12.7Hz,2H),1.60(s,2H),1.39–0.97(m,10H).

[0391] Embodiment 14

[0392]

[0393] In a 50mL single-mouth bottle, formic acid (2.44g, 53.009mmol) and acetic anhydride (2.174g, 21.295mmol) were refluxed for 1h, and then intermediate 1 (0.100g, 0.259mmol) was added to the single-mouth bottle, and the reflux reaction was continued for 2h. After the reaction was completed, the crude product was directly freeze-dried using a freeze dryer, and the crude product was purified by preparative liquid phase to obtain compound 13 (20.00mg, yield: 18.63%).

[0394] LC-MS:415.2[M+H] + .

[0395] 1 H NMR(400MHz,TFA-d1)δ9.80–9.32(m,1H),8.86(s,1H),7.75(d,J=5.4Hz,1H),7.59(d ,J=7.8Hz,1H),7.48(d,J=5.5Hz,1H),7.37(t,J=7.7Hz,1H),7.01(d,J=7.6Hz,1H),6 .61(s,1H),3.46(s,5H),2.62(t,J=4.6Hz,4H),2.43(t,J=7.5Hz,2H),1.98–1.86(m, 2H),1.84–1.73(m,2H),1.43(q,J=7.1Hz,2H),1.33–1.14(m,3H),1.12–0.96(m,2H).

[0396] Embodiment 15

[0397]

[0398] Intermediate 1 (0.100 g, 0.259 mmol) was dissolved in tetrahydrofuran (6 mL), pyridine (0.098 g, 1.239 mmol) and intermediate 10 (48.612 mg, 0.26 mmol) were added, and the mixture was reacted at 50 °C for 3 h. The reaction solution was diluted with water and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered and dried by rotation. The crude product was subjected to thin layer chromatography to obtain compound 14 (30.5 mg, yield: 13.27%).

[0399] LC-MS: 887.35[M+H] + .

[0400] 1H NMR (400MHz, DMSO-d6) δ10.05 (s, 2H), 7.70 (d, J = 8.0Hz, 2H), 7.55-7.51 (m, 4 H),7.43(d,J=8Hz,2H),7.34-7.30(m,2H),6.06(d,J=8.0Hz,2H),4.27(s,4H) ,3.45(s,2H),3.13(s,8H),2.57(s,8H),2.40-2.36(m,4H),1.88-1.77(m,8H) ,1.41-1.36(m,4H),1.30-1.23(m,2H),1.20-1.14(m,4H),1.05-0.97(m,4H).

[0401] Example 16

[0402]

[0403] Intermediate 1 (0.201 g, 0.52 mmol) was dissolved in tetrahydrofuran (5 mL), pyridine (0.411 mg, 5.2 mmol) and intermediate 4 (0.670 g, 5.2 mmol) were added, and the mixture was reacted at 40 ° C for 3 h. The reaction solution was cooled and filtered, and the solid was collected and washed twice with tetrahydrofuran. The crude product was purified by column chromatography to obtain compound 15 (23.1 mg, yield: 9.27%).

[0404] LC-MS:479.15[M+H] + .

[0405] 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),9.57(s,1H),7.54(d,J=8.0Hz,1H),7. 59(d,J=8.0Hz,1H),7.46-7.42(m,2H),7.37-7.33(m,1H),7.01(d,J=8.0Hz, 1H),5.90-5.76(m,2H),5.29(s,1H),3.46(s,1H),3.24(s,3H),2.96-2.79(m ,5H),1.87-1.72(m,4H),1.50(s,2H),1.28-1.18(m,4H),1.08-0.98(m,3H).

[0406] Embodiment 17

[0407]

[0408] Intermediate 1 (0.100 g, 0.259 mmol) was dissolved in tetrahydrofuran (6 mL), pyridine (0.098 g, 1.239 mmol) and intermediate 11 (159.250 mg, 1.3 mmol) were added, and the reaction was carried out at room temperature for 3 h. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by column chromatography to obtain compound 16 (58.9 mg, yield: 48.12%).

[0409] LC-MS:473.05[M+H] + .

[0410] 1 H NMR (400MHz, DMSO-d6) δ10.55(s,1H),7.77(d,J=4.0Hz,1H),7.63(d,J=8.0Hz,1H),7.48(d,J=4.0Hz,1H),7.40-7.36(m,1H),7.05( d,J=8.0Hz,1H),3.78(s,3H),3.66-3.54(m,3H),3.19(s,8H),1.89-1.75(m,4H),1.61(s,2H),1.32-1.18(m,4H),1.09-0.99(m,2H).

[0411] Embodiment 18

[0412]

[0413] In a sealed tube, intermediate 1 (0.300 g, 0.777 mmol) was dissolved in DMF (3 mL), intermediate 12 (0.418 g, 1.56 mmol), potassium phosphate (0.332 g, 1.56 mmol) and cuprous iodide (15 mg, 0.08 mmol) were added, and the reaction was carried out at 90 ° C for 12 h. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was prepared by reverse phase to obtain compound 17 (12.8 mg, yield:

[0414] 2.10%), as a white solid.

[0415] LC-MS: 785.45[M+H] + .

[0416] 1H NMR (400MHz, DMSO-d6) δ7.71(d,J=4.0Hz,2H),7.54(d,J=8.0Hz,2H),7.43(d,J=8.0Hz,2H),7.34-7.30(m,2H),6.96(d,J=8.0Hz,2H),6.26(s,1H),5 .98(s,1H),5.32(s,1H),4.21(s,2H),3.13(s,8H),2.57(s,8H),2.39-2.3 4(m,4H),1.99(s,2H),1.78-1.71(m,8H),1.36(s,4H),1.05-0.95(m,8H).

[0417] Embodiment 19

[0418]

[0419] In a sealed tube, intermediate 1 (0.100 g, 0.259 mmol) and paraformaldehyde (117 mg, 1.3 mmol) were dissolved in tetrahydrofuran (2 mL) and water (1 mL) and reacted overnight at 80° C. The reaction solution was concentrated and the crude product was prepared by reverse phase to give compound 18 (45.8 mg, yield: 39.59%).

[0420] LC-MS:447.15[M+H] + .

[0421] 1 H NMR (400MHz, DMSO-d6) δ7.71(d,J=4.0Hz,1H),7.54(d,J=8.0Hz,1H),7.43(d,J=8.0Hz,1H),7.34-7. 30(m,1H),6.96(d,J=8.0Hz,1H),6.32-6.28(m,0.7H),5.90-5.85(m,1H),5.54-5.50(m,0.6H),5.21 -5.18(m,0.7H),4.64(d,J=8.0Hz,1H),4.44-4.41(m,1.6H),3.30-3.27(m,1H),3.13(s,4H),2.57(s ,4H),2.39-2.36(m,2H),1.82-1.72(m,4H),1.38-1.35(m,2H),1.23-1.16(m,1H),1.10-0.96(m,4H).

[0422] Embodiment 20

[0423]

[0424] In a sealed tube, intermediate 1 (0.050 g, 0.130 mmol) and paraformaldehyde (35 mg, 0.39 mmol) were dissolved in ethanol (3 mL), 1N sodium hydroxide aqueous solution (0.02 mL) was added, and the reaction was continued at 80 ° C overnight. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and dried. The crude product was purified by column chromatography to obtain compound 19 (9.8 mg, yield: 16.95%).

[0425] LC-MS:445.20[M+H] + .

[0426] 1 H NMR (400MHz, DMSO-d6) δ7.71(d,J=8.0Hz,1H),7.55(d,J=8.0Hz,1H),7.44(d,J=4.0Hz,1 H),7.35-7.31(m,1H),6.97(d,J=8.0Hz,1H),6.55-6.50(m,1H),5.89(d,J=8.0Hz,1H),4 .44(d,J=8.0Hz,2H),3.39-3.35(m,3H),3.14(s,4H),2.58(s,4H),2.39(s,2H),1.82(d, J=8.0Hz,2H),1.74(d,J=12.0Hz,2H),1.38(s,2H),1.28-1.26(m,1H),1.08-0.96(m,7H).

[0427] Embodiment 21

[0428]

[0429] In a sealed tube, intermediate 1 (0.050 g, 0.130 mmol) and paraformaldehyde (35 mg, 0.39 mmol) were dissolved in ethanol (3 mL) and reacted at 80°C overnight. The reaction solution was diluted with water and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was prepared using reverse phase to obtain compound 20 (15.5 mg, yield: 28.62%).

[0430] LC-MS:417.20[M+H] + .

[0431] 1H NMR (400MHz, DMSO-d6) δ8.20(s,1H),7.71(d,J=8.0Hz,1H),7.54(d,J=8.0Hz,1H ),7.44(d,J=4.0Hz,1H),7.34-7.30(m,1H),6.96(d,J=8.0Hz,1H),6.33-5.86(m ,1H),5.30-4.42(m,2H),3.32-3.26(m,1H),3.14(s,4H),2.58(s,4H),2.41-2.3 7(m,2H),1.82-1.72(m,4H),1.37-1.35(m,2H),1.23(s,1H),1.07-0.95(m,4H).

[0432] Embodiment 22

[0433]

[0434] Compound a (0.116 g, 0.3 mmol) was dissolved in dichloromethane (5 mL), triethylamine (0.243 g, 2.4 mmol) and triphosgene (0.045 g, 0.15 mmol) were added at 0°C, reacted at room temperature for half an hour, and intermediate 13 (50 mg, 0.6 mmol) was added, and reacted at room temperature for half an hour. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by column chromatography to obtain compound 21 (22.1 mg, yield: 17.68%).

[0435] LC-MS:417.15[M+H] + .

[0436] 1 H NMR (400MHz, DMSO-d6) δ8.94(s,1H),7.71(d,J=4.0Hz,1H),7.54(d,J=8.0Hz,1H) ,7.44(d,J=4.0Hz,1H),7.34-7.31(m,1H),6.97(d,J=4.0Hz,1H),6.53(d,J=8.0Hz ,1H),3.50(s,3H),3.42-3.38(m,1H),3.14(s,4H),2.58(s,4H),2.40-2.37(m,2H) ,1.75(d,J=12.0Hz,4H),1.40-1.35(m,2H),1.29-1.23(m,3H),1.02-1.95(m,2H).

[0437] Embodiment 23

[0438]

[0439] Compound a (0.170 g, 0.44 mmol) was dissolved in dichloromethane (5 mL), triethylamine (0.354 g, 3.5 mmol) and triphosgene (0.078 g, 0.264 mmol) were added at 0°C, and the reaction was carried out at room temperature for half an hour. Intermediate 14 (0.066 g, 0.88 mmol) was added, and the reaction was carried out at room temperature for half an hour. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was subjected to column chromatography to obtain compound 22 (18.2 mg, yield: 9.30%).

[0440] LC-MS:445.15[M+H] + .

[0441] 1 H NMR (400MHz, DMSO-d6) δ7.71(d,J=4.0Hz,1H),7.55(d,J=8.0Hz,1H),7.43( d,J=4.0Hz,1H),7.34-7.31(m,1H),6.97(d,J=8.0Hz,1H),5.77-5.71(m,2H ),3.30-3.27(m,2H),3.23(s,3H),3.14-3.10(m,6H),2.57(s,4H),2.38(s, 2H),1.81-1.72(m,4H),1.37-1.35(m,2H),1.23(s,2H),1.04-0.94(m,4H).

[0442] Embodiment 24

[0443]

[0444] Step 1: Preparation of intermediate 16

[0445] Intermediate 6 (2.0 g, 8.13 mmol) was dissolved in DMF (10 mL), and intermediate 15 (937 mg, 9.75 mmol) was added, and the mixture was reacted at room temperature for 2 h. The reaction solution was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and vortexed to obtain intermediate 16.

[0446] 1 H NMR (400MHz, CDCl3) δ5.81(s,2H),2.92-2.87(m,2H),2.43-2.37(m,2H),1.34-1.31(m,3H0,1.18-1.14(m,3H).

[0447] Step 2: Preparation of intermediate 17

[0448] Intermediate 16 (1 g, 5.2 mmol) was dissolved in dichloromethane (1 mL), and sulfonyl chloride (1 g, 7.8 mmol) was added at 0°C, and the mixture was reacted at room temperature for 2 h. After the reaction, the reaction solution was dried to obtain intermediate 17, which was used in the next step.

[0449] Step 3: Preparation of compound 23

[0450] Intermediate 1 (200 mg, 0.52 mmol) was dissolved in tetrahydrofuran (5 mL), pyridine (0.1 mL) and intermediate 17 (600 mg, 3.6 mmol) were added, and the reaction was carried out at room temperature for 16 h. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and dried by rotation. The crude product was subjected to thin layer chromatography to obtain compound 23 (27.6 mg, yield: 10.3%).

[0451] LC-MS: 517.25[M+H] + .

[0452] 1 H NMR (400MHz, DMSO-d6): δ10.26(s,1H),7.71(d,J=4.0Hz,1H),7.54(d,J=8 .0Hz,1H),7.46-7.42(m,2H),7.32-7.30(m,1H),6.97(d,J=4.0Hz,1H),5.7 0(s,2H),3.50-3.42(m,1H),3.14(s,4H),2.58(s,4H),2.41-2.36(m,4H),1 .87-1.75(m,4H),1.41-1.36(m,2H),1.23-1.16(m,4H),1.04-1.00(m,4H).

[0453] Embodiment 25

[0454]

[0455] Step 1: Preparation of intermediate 19

[0456] Intermediate 6 (2.0 g, 8.13 mmol) was dissolved in DMF (20 mL), and intermediate 19 (980 mg, 8.94 mmol) was added, and the reaction was carried out at room temperature for 2 h. After the reaction was completed, water was added to dilute and extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was subjected to column chromatography to obtain intermediate 19 (1.4 g, yield: 82.6%).

[0457] 1H NMR (400MHz, CDCl3) δ5.81 (s, 2H), 2.92-2.86 (m, 2H), 2.64-2.57 (m, 1H), 1.34-1.31 (m, 3H), 1.19 (d, J = 8.0Hz, 6H).

[0458] Step 2: Preparation of intermediate 20

[0459] Intermediate 19 (1.4 g, 6.80 mmol) was dissolved in DCM (2 mL), and sulfonyl chloride (1.1 g, 8.16 mmol) was added at 0°C. The reaction was carried out at room temperature for 3 h. After the reaction was completed, the solvent was dried to obtain a crude intermediate 20 (900 mg, yield: 75%).

[0460] 1 H NMR (400MHz, CDCl3) δ5.83 (s, 2H), 2.68-2.59 (m, 1H), 1.22-1.18 (m, 6H).

[0461] Step 3: Preparation of compound 24

[0462] Intermediate 1 (200 mg, 0.52 mmol) was dissolved in THF (5 mL), pyridine (0.1 mL) was added, and intermediate 20 (280 mg, 1.55 mmol) was added under ice bath, and the reaction was carried out at room temperature for 12 h. After the reaction was completed, water was added to dilute and extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and dried. The crude product was purified by column chromatography to obtain compound 24 (38.4 mg, yield: 13.9%).

[0463] LC-MS: 531.35[M+H] + .

[0464] 1 H NMR (400MHz, DMSO-d6) δ10.26(s,1H),7.70(d,J=4.0Hz,1H),7.54(d,J=8.0Hz,1H ),7.45-7.43(m,2H),7.34-7.30(m,1H),6.97(d,J=4.0Hz,1H),5.71(s,2H),3.48- 3.42(m,1H),3.14(s,4H),2.59-2.55(m,4H),2.41-2.38(m,2H),1.87-1.74(m,4H ),1.41-1.36(m,2H),1.28-1.14(m,4H),1.09(d,J=8.0Hz,6H),1.04-0.98(m,2H).

[0465] Embodiment 26

[0466]

[0467] Step 1: Preparation of intermediate 22

[0468] Intermediate 21 (12.0 g, 83.93 mmol) was dissolved in methyl tert-butyl ether (100 mL), DIEA (11.9 g, 92.32 mmol) was added, and compound 2 (5.5 g, 88.12 mmol) was added under ice bath, and the reaction was carried out at room temperature for 16 h. After the reaction was completed, 1N HCl (50 mL) was added to quench the reaction, the reaction solution was extracted with methyl tert-butyl ether, and the organic phase was washed with 1N HCl (50 mL) and saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried to obtain intermediate 22 (11.0 g, yield: 77.7%).

[0469] 1 H NMR (400MHz, CDCl3) δ6.62-6.58(m,1H),2.94-2.88(m,2H),1.81(d,J=8.0Hz,3H),1.36-1.32(m,3H).

[0470] Step 2: Preparation of intermediate 23

[0471] Intermediate 22 (3.0 g, 17.79 mmol) was dissolved in acetic acid (5 mL), DIEA (6.9 g, 53.37 mmol) was added, and the mixture was reacted at 55°C for 16 h. After the reaction was completed, the reaction solution was diluted with water and extracted with ethyl acetate, and the organic phase was washed with saturated sodium carbonate aqueous solution and brine, dried over anhydrous sodium sulfate, filtered and dried to obtain intermediate 23 (3.3 g, yield: 97.0%).

[0472] 1 H NMR (400MHz, CDCl3) δ6.94-6.90(m,1H),2.91-2.81(m,2H),2.07(s,3H),1.49(d,J=8.0Hz,3H),1.32-1.29(m,3H).

[0473] Step 3: Preparation of intermediate 24

[0474] Intermediate 23 (3.3 g, 17.17 mmol) was dissolved in dichloromethane (3 mL), and sulfonyl chloride (3.5 g, 25.75 mmol) was added under ice bath. The mixture was reacted at room temperature for 3 h. After the reaction, the solvent was dried to obtain intermediate 24 (2.1 g, yield: 73.4%), which was used in the next step.

[0475] 1H NMR (400MHz, CDCl3) δ6.82-6.79 (m, 1H), 2.12 (s, 3H), 1.58 (d, J = 4.0Hz, 3H).

[0476] Step 4: Preparation of compound 25

[0477] Intermediate 1 (600 mg, 1.55 mmol) was dissolved in dichloromethane (10 mL), pyridine (0.1 mL) and intermediate 24 (2.0 g, 12.0 mmol) were added under ice bath, and the reaction was carried out at room temperature for 16 h. After the reaction was completed, water was added to quench the reaction, and dichloromethane was extracted. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was subjected to thin layer chromatography to obtain compound 25 (11.6 mg, yield: 1.4%).

[0478] LC-MS: 517.25[M+H] + .

[0479] 1 H NMR (400MHz, DMSO-d6) δ10.18(s,1H),7.71(d,J=4.0Hz,1H),7.54(d,J=8.0Hz,1H),7.46- 7.42(m,2H),7.34-7.30(m,1H),6.96(d,J=8.0Hz,1H),6.72-6.68(m,1H),3.49-3.45(m,1 H),3.13(s,4H),2.57(s,4H),2.40-2.36(m,2H),2.04(s,3H),1.85(s,2H),1.76(d,J=12. 0Hz, 2H), 1.43 (d, J = 4.0Hz, 3H), 1.39-1.35 (m, 2H), 1.23-1.19 (m, 3H), 1.05-0.98 (m, 2H).

[0480] Embodiment 27

[0481]

[0482] Step 1: Preparation of intermediate 26

[0483] Intermediate 6 (6.0 g, 24.38 mmol) was dissolved in DMF (50 mL), and intermediate 25 (3.1 g, 29.25 mmol) was added, and the reaction was carried out at room temperature for 16 h. The reaction solution was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and dried. The crude product was purified by column chromatography to obtain intermediate 26 (3.6 g, yield: 72.3%).

[0484] 1H NMR (400MHz, CDCl3) δ5.80(s,2H),2.92-2.86(m,2H),1.65-1.62(m,1H),1.34-1.31(m,3H),1.09-1.05(m,2H),0.96-0.91(m,2H).

[0485] Step 2: Preparation of intermediate 27

[0486] Intermediate 26 (1 g, 4.89 mmol) was dissolved in dichloromethane (1 mL), and sulfuryl chloride (989 mg, 7.33 mmol) was added at 0°C and reacted at room temperature for 2 h. After the reaction, the reaction solution was dried to obtain intermediate 27 (700 mg, yield: 80.2%), which was used in the next step.

[0487] Step 3: Preparation of compound 26

[0488] Intermediate 1 (200 mg, 0.52 mmol) was dissolved in tetrahydrofuran (5 mL), pyridine (0.1 mL) and intermediate 27 (700 mg, 3.92 mmol) were added, and the reaction was carried out at room temperature for 16 h. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and dried by rotation. The crude product was subjected to thin layer chromatography to obtain compound 26 (56.3 mg, yield: 20.5%).

[0489] LC-MS: 529.15[M+H] + .

[0490] 1 H NMR (400MHz, DMSO-d6): δ10.27(s,1H),7.71(d,J=4.0Hz,1H),7.54(d,J=8.0Hz,1H),7.45-7 .42(m,2H),7.34-7.30(m,1H),6.97(d,J=4.0Hz,1H),5.70(s,2H),3.49-3.42(m,1H),3.14(s ,4H),2.58(s,4H),2.40-2.37(m,2H),1.87(d,J=12.0Hz,2H),1.77((d,J=12.0Hz,2H),),1. 72-1.65(m,1H),1.41-1.36(m,2H),1.23-1.17(m,5H),0.98-0.96(m,2H),0.90-0.88(m,2H).

[0491] Embodiment 28

[0492]

[0493] Step 1: Preparation of intermediate 29

[0494] Intermediate 6 (2 g, 8.16 mmol) was dissolved in DMF (20 mL) and added to intermediate 28 (1.1 g, 9.8 mmol), and the mixture was reacted at room temperature for 2 h. The reaction solution was diluted with water and extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by column chromatography to obtain intermediate 29 (1 g, yield: 59.5%).

[0495] 1 H NMR (400MHz, CDCl3) δ5.77(s,2H),2.90-2.85(m,2H),2.38-2.34(m,2H),1.59-1.50(m,2H),1.26-1.22(m,3H),0.89-0.86(m,3H).

[0496] Step 2: Preparation of intermediate 30

[0497] Intermediate 29 (1 g, 4.85 mmol) was dissolved in dichloromethane (1 mL), and sulfuryl chloride (780 mg, 5.82 mmol) was added at 0°C, and the mixture was reacted at room temperature for 2 h. The reaction solution was dried by rotary evaporation to obtain intermediate 30 (800 mg, yield: 91.5%), which was used in the next step.

[0498] 1 H NMR (400MHz, CDCl3) δ5.83(s,2H),2.42-2.38(m,2H),1.72-1.67(m,2H),0.99-0.95(m,3H).

[0499] Step 3: Preparation of compound 27

[0500] Intermediate 1 (200 mg, 0.52 mmol) was dissolved in tetrahydrofuran (20 mL), pyridine (123 mg, 1.56 mmol) and intermediate 30 (468 mg, 2.6 mmol) were added, and the reaction was carried out at room temperature for 2 h. The reaction solution was diluted with water and extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried. The crude product was purified by column chromatography to obtain compound 27 (20.7 mg, yield: 7.5%).

[0501] LC-MS: 531.35[M+H] + .

[0502] 1H NMR (400MHz, DMSO-d6) δ10.29(s,1H),7.71(d,J=4.0Hz,1H),7.54(d,J=8.0Hz,1H),7.47-7.42(m,2H),7.34-7.30(m,1H),6 .96(d,J=8.0Hz,1H),5.70(s,2H),3.49-3.44(m,1H),3.14(s,4H),2.57(s,4H),2.40-2.33(m,4H),2.15-2.12(m,1H),1.87 -1.75(m,4H),1.57-1.47(m,2H),1.41-1.36(m,2H),1.23-1.16(m,2H),1.05-0.99(m,2H),0.90-0.86(m,3H).

[0503] Embodiment 29

[0504]

[0505] Step 1: Preparation of intermediate 32

[0506] Intermediate 6 (2.0 g, 8.13 mmol) was dissolved in dichloromethane (50 mL) and water (10 mL), and sodium bicarbonate (1.8 g, 10.89 mmol) and tetrabutylammonium hydrogen sulfate (3.7 g, 10.89 mmol) were added. After reacting at room temperature for 1 h, intermediate 31 (1.4 g, 10.89 mmol) was dissolved in dichloromethane and added dropwise to the above reaction solution. After reacting at room temperature for 2 h, the reaction solution was diluted with water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried. The crude product was mixed and purified by column chromatography to obtain intermediate 32 (1.4 g, yield: 69.9%).

[0507] 1 H NMR (400MHz, CDCl3) δ5.79 (s, 2H), 2.88 (q, J = 8.0 16.0Hz,2H),2.31-2.35(m,1H),1.90(d,J=13.0Hz,2H),1.74(d,J=12.2Hz,2H),1.64(s,1H),1.49-1.40(m,2H),1.34–1.22(m,6H).

[0508] Step 2: Preparation of intermediate 33

[0509] Intermediate 32 (1 g, 4.06 mmol) was dissolved in dichloromethane (1 mL), and sulfonyl chloride (0.8 g, 6.09 mmol) was added dropwise under ice bath. After the addition, the reaction solution was reacted at room temperature for 2 h. After the reaction, the reaction solution was directly spin-dried to obtain intermediate 33, and the crude product was directly used in the next step.

[0510] 1 H NMR (400MHz, CDCl3) δ5.81 (s, 2H), 2.35-2.38 (m, 1H), 1.92 (d, J = 13.2Hz, 1H) ,1.80-1.72(m,1H),1.67-1.61(m,1H),1.51-1.40(m,1H),1.34-1.20(m,1H).

[0511] Step 3: Preparation of compound 28

[0512] Intermediate 1 (100 mg, 0.25 mmol) was dissolved in THF (10 mL), and pyridine (102.3 mg, 1.29 mmol) was added. Intermediate 33 (345.0 mg, 1.29 mmol) was added under ice bath. The reaction solution was reacted at room temperature for 16 h. The reaction solution was directly spin-dried, and the crude product was purified by thin layer chromatography to obtain compound 28 (51.2 mg, yield: 34.6%).

[0513] LC-MS: 571.45 [M+H] + .

[0514] 1 H NMR (400MHz, CD3OD) δ7.54-7.50(m,2H),7.33-7.29(m,2H),6.99(d,J=7.7Hz,1H),5.75(s,2H),3.57-3.55(m,1H),2.93-2.90(m,4H),2.68-2.67( m,2H),2.38-2.36(m,1H),2.00(d,J=10.4Hz,2H),1.88-1.87(m,4H),1.7 3-1.72(m,2H),1.66-1.64(m,1H),1.54-1.52(m,2H),1.49-1.04(m,14H).

[0515] Embodiment 30

[0516]

[0517] Compound 15 (50 mg, 0.10 mmol) was dissolved in tetrahydrofuran (5 mL), and aqueous ammonia (5 mL) was added. The reaction was allowed to react at room temperature for 16 h. The reaction solution was diluted with water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried. The crude product was purified using a thin layer chromatography plate to obtain compound 29 (15.4 mg, yield: 34.3%).

[0518] 1 H NMR (400MHz, CD3OD-d6) δ7.50(t,J=7.1Hz,2H),7.32-7.28(m,2H),6.96(d,J=7.6Hz,1H),3.50-3.49(m,1H),3.23(s,4H),2 .73-2.72(m,4H),2.54-2.49(m,2H),1.97(d,J=10.8Hz,2H),1.84(d,J=11.6Hz,2H),1.51-1.49(m,2H),1.32-1.06(m,7H).

[0519] Embodiment 31

[0520]

[0521] Step 1: Preparation of intermediate 2

[0522] Intermediate 1 (200 mg, 0.52 mmol) was dissolved in dichloromethane (20 mL), pyridine (409 mg, 5.2 mmol) was added, and chloroacetyl chloride (584 mg, 5.2 mmol) was added dropwise under ice bath, and the mixture was reacted at room temperature for 1 h. The reaction solution was directly spin-dried, the crude product was mixed, and intermediate 2 (200 mg, yield: 83.5%) was obtained by column chromatography as a brown solid.

[0523] Step 2: Preparation of intermediate 34

[0524] Dissolve intermediate 2 (200 mg, 0.43 mmol) in tetrahydrofuran (5 mL), add ammonia water (5 mL), and react at room temperature for 16 h. The reaction raw material consumption is 30%, and the reaction solution is diluted with water, saturated sodium bicarbonate solution is appropriate, and extracted with ethyl acetate. The organic phases are combined, washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried. The crude product is purified by thin layer chromatography to obtain intermediate 34 (50 mg, yield: 26.1%).

[0525] Step 3: Preparation of compound 30

[0526] Intermediate 34 (50 mg, 0.11 mmol) was dissolved in dichloromethane (10 mL), pyridine (89.2 mg, 1.1 mmol) was added, and acetyl chloride (88.6 mg, 1.1 mmol) was added dropwise under ice bath. After the addition, the reaction was allowed to react at room temperature for 1 h. The reaction solution was diluted with water and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and then spin-dried. The crude product was subjected to thin layer chromatography to obtain compound 30 (32.3 mg, yield: 59.0%).

[0527] LC-MS:486.35[M+H] + .

[0528] 1 H NMR (400MHz, CD3OD) δ7.54-7.45(m,2H),7.34-7.27(m,2H),6.97(d,J=7.6Hz,1H),4.57(s,2H),3.90(s,2H),3.61-3.51(m,1H),3.23(s,4H) ,2.72(s,4H),2.55–2.45(m,2H),1.99(s,5H),1.84(d,J=12.8Hz,2H), 1.52-1.48(m,2H),1.33(s,1H),1.23-1.21(m,1H),1.10-0.88(m,2H).

[0529] Embodiment 32

[0530]

[0531] Step 1: Preparation of intermediate 36

[0532] Intermediate 5 (5 g, 20.3 mmol) was dissolved in N,N-dimethylformamide (100 mL), and intermediate 35 (7.57 g, 60.9 mmol) was added. The reaction solution was reacted at room temperature for 16 h. The reaction solution was diluted with water, extracted three times with ethyl acetate, and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried. The crude product was mixed and intermediate 36 (3.4 g, yield: 47.7%) was obtained by column chromatography.

[0533] 1 H NMR (400MHz, CDCl3) δ5.80 (s, 2H), 2.88 (q, J=8.0 12.0Hz, 2H), 1.31 (t, J=8.0Hz, 3H), 1.21 (s, 9H).

[0534] Step 2: Preparation of intermediate 37

[0535] Intermediate 36 (500 mg, 2.26 mmol) was dissolved in dichloromethane (1 mL), and sulfonyl chloride (454 mg, 3.4 mmol) was added dropwise under ice bath. After the addition, the mixture was reacted at room temperature for 1 h. After the reaction was complete, the reaction solution was dried at room temperature to obtain intermediate 37, and the crude product was directly used in the next step.

[0536] Step 3: Preparation of compound 31

[0537] Intermediate 1 (150 mg, 0.388 mmol) was dissolved in dichloromethane (10 mL), pyridine (306 mg, 3.88 mmol) was added, and intermediate 37 (442 mg, 2.27 mmol) was added under ice bath, and the reaction was carried out at room temperature for 16 h. The reaction solution was diluted with water and extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate and then spin-dried, and the crude product was purified by thin layer chromatography to obtain compound 31 (39.5 mg, yield: 18.6%).

[0538] LC-MS: 545.35[M+H] + .

[0539] 1 H NMR (400MHz, CDCl3) δ7.52(d,J=7.9Hz,1H),7.44(d,J=7.7Hz,1H),7.40(d,J=5.4Hz,1H), 7.35-7.29(m,1H),7.03(s,1H),6.94(d,J=7.6Hz,1H),5.77(s,2H),3.63-3.61(m,1H),3. 34-3.32(m,4H),2.75-2.73(m,4H),2.53-2.52(m,2H),2.03(d,J=12.1Hz,2H),1.82(d,J= 11.5Hz,2H),1.50-1.48(m,2H),1.25-1.24(m,2H),1.22-1.20(m,9H),1.18-1.03(m,3H).

[0540] Embodiment 33

[0541]

[0542] Step 1: Preparation of intermediate 39

[0543] Intermediate 5 (6 g, 35.7 mmol) was dissolved in N,N-dimethylformamide (50 mL), and intermediate 38 (5.9 g, 53.56 mmol) was added at room temperature. The reaction solution was heated at 50°C for 16 h. The reaction solution was diluted with water, extracted three times with ethyl acetate, and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried. The crude product was mixed and intermediate 39 (3.4 g, yield: 47.6%) was obtained by column chromatography.

[0544] 1 H NMR (400MHz, CDCl3) δ6.92 (q, J=4.0 12.0Hz,1H),2.89-2.82(m,2H),2.57-2.50(m,1H),1.49(d,J=5.5Hz,3H),1.30(t,J=7.4Hz,3H),1.17(d,J=7.0Hz,6H).

[0545] Step 2: Preparation of intermediate 40

[0546] Intermediate 39 (800 mg, 3.362 mmol) was dissolved in dichloromethane (1 mL), and sulfonyl chloride (736 mg, 5.448 mmol) was added dropwise under ice bath, and the mixture was reacted at room temperature for 1 h. After the reaction was complete, the reaction solution was dried at room temperature to obtain intermediate 40, and the crude product was directly used in the next step.

[0547] Step 3: Preparation of compound 32

[0548] Intermediate 1 (100 mg, 0.26 mmol) was dissolved in THF (10 mL), pyridine (205 mg, 2.6 mmol) was added, and intermediate 40 (400 mg, 2.0 mmol) was added under ice bath, and the reaction solution was reacted at room temperature for 16 h. The reaction solution was diluted with water and extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and then spin-dried. The crude product was purified using a thin layer chromatography plate to obtain compound 32 (4.2 mg, yield: 2.9%).

[0549] LC-MS: 545.20 [M+H] + .

[0550] 1H NMR(400MHz,CD3OD)δ7.52-7.50(m,2H),7.35 -7.27(m,2H),6.96(d,J=7.6Hz,1H),6.78(t,J=5.4Hz,1H),3.57-3.56(m,1H),3.22(s,4H),2.71(s,4H),2.52-3.50 (m,3H),1.98(d,J=12.4Hz,2H),1.84(d,J=12.9Hz,2H),1.47-1.45(m,3H),1.25-1.20(m,6H),1.13(d,J=7.0Hz,6H).

[0551] Embodiment 34

[0552] Preparation of N'-trans-4-[2-[4-(Benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-3-methylurea (Compound c)

[0553]

[0554] Dissolve 20g of trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexylamine (compound a) in 160mL of dichloromethane, add 13.2g of CDI (N,N'-carbonyldiimidazole), cool to -10℃-0℃, drop 29.2g of triethylamine, and react at this temperature for 1h. Then add 9.73g of methylamine hydrochloride, and react at room temperature (25℃±2℃) overnight. After the reaction, add 50mL of water and stir for 30-60min, filter, and dry to obtain 13.2g of compound c.

[0555] 1 H NMR (400MHz, d-TFA) δ8.387(d,J=8Hz,1H),7.95-7.83(m,4H),5.12(t,J=12Hz,2H),4.72-4.60(m,4H),4.44(t,J=12.4Hz ,2H),3.93-3.83(m,3H),3.26(s,3H),2.43(d,J=10.8Hz,2H),2.23-2.14(m,4H),1.77-1.69(m,3H),1.542-1.453(m,2H).

[0556] 13C-NMR(101MHz,d-TFA)δ158.809,144.685,132.931,129.466,127.652,125.771,125.668,1 25.404,117.229,56.969,51.782,50.950,49.732,33.961,31.363,30.478,30.175,26.891.

[0557] Embodiment 35

[0558] Preparation of N'-trans-4-[2-[4-(Benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea maleate (Compound e)

[0559]

[0560] Step 1: Preparation of compound d

[0561] Dissolve 1.73 g of trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexylamine (compound a) in 50 mL of dichloromethane, add 1.40 mL of triethylamine, and then add 5.50 mmol of N,N-diformylformyl chloride. Stir at room temperature (25°C±2°C) for 48 hours. After the reaction is completed, add 50 mL of water to extract the liquid, concentrate the organic phase (0.01 MPa, 45°C), collect the target component by column chromatography (400 mesh silica gel model) with methanol: dichloromethane = 1:10, and concentrate to obtain 1.89 g of amorphous compound d.

[0562] 1 H-NMR (CDC13) ppm: 7.79 (1H, d, J = 5.5Hz), 7.76 (1H, d, J = 8.1Hz), 7.33 (1H, m), 7.28 (1H, d, 7.6Hz), 6.96 (1H, d, J = 7.6Hz), 6.48 (1H,brs),3.44-3.36(8H,m),3.58(1H,m),3.01(6H,s),2.46(2H,m),1.68-1.42(8H,m),1.52-1.48(1H,m),1.38-1.36(2H,m).

[0563] Step 2: Preparation of compound e

[0564] Dissolve 15g of N'-trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound d) in 67.5g of anhydrous ethanol, add 4.7g of anhydrous ethanol solution of maleic acid dropwise at 45°C-50°C, and keep warm for 1.5 hours. Slowly return to room temperature under stirring, solid precipitates, filter, wash the filter cake twice with anhydrous ethanol, and dry under vacuum to obtain 12.8g of Compound e.

[0565] Biological Example 1 Receptor Binding Assay

[0566] 1. Purpose of the experiment

[0567] The main purpose of this test is to determine the effect of compounds on 5-hydroxytryptamine receptors (5-HT 1A , 5-HT 2A and 5-HT 2B )'s affinity K i .

[0568] 2. Experimental Materials and Instruments

[0569] 2.1 Test products

[0570] Preparation method: Dissolve the test compounds in DMSO to make 10mM stock solution and store in a 4℃ refrigerator.

[0571] 2.2 Receptor membrane proteins

[0572] Expresses 5-HT receptors 1A , 5-HT 2A and 5-HT 2B ) The information of three cell membrane proteins is as follows:

[0573] Table 1. 5-HT receptor membrane proteins

[0574] Receptors Gene Abbreviation Genebank accession number Host cells Storage concentration (mg / mL) 5-HT1A HTR1A NM_000524 CHO 0.8 5-HT2A HTR2A NM_000621 HEK293 1.95 5-HT2B 5-HT2B NM_000867 HEK293 7

[0575] 2.3 Reagents and consumables

[0576] Table 2. Reagents and consumables

[0577]

[0578]

[0579] 2.4 Main instruments

[0580] Table 3. Instruments and equipment

[0581] Instrument No. instrument model Manufacturer 834638 MicroBeta2 2450Microplate Counter PerkinElmer 256866 Harvest C961961 PerkinElmer 300943 Shaker 4625-1CECN Thermo Oven No. 1 Oven HPX-9052MBE BOXUN 401242 Ultrapure water instrument Milli-Q Millipore

[0582] 3. Experimental Methods

[0583] 1) Configuration of cell membrane

[0584] 5-HT 1A 5-HT was added to the assay buffer. 1A The membrane was diluted to 100 μg / mL.

[0585] 5-HT 2A 5-HT was added to the assay buffer. 2A The membrane was diluted to 50 μg / mL.

[0586] 5-HT 2B 5-HT was added to the assay buffer. 2B The membrane was diluted to 50 μg / mL.

[0587] 2) Isotope configuration

[0588] 5-HT 1A : [3H]8-OH-DPAT was diluted to 0.6 nM in assay buffer.

[0589] 5-HT 2A : Dilute [3H]-Ketanserin to 2 nM in assay buffer.

[0590] 5-HT 2B : Dilute [3H]-LSD to 2 nM in assay buffer.

[0591] 3) Make 8 4-fold dilutions of the test compound and reference compound, and add 1 μL to each well.

[0592] 5-HT 1A The initial concentration of the reference compound 8-OH-DPAT was 1000 nM; for 5-HT 2A The initial concentration of the reference compound Ketanserin was 1000 nM; for 5-HT 2B , reference compound

[0593] ( + )The starting concentration of DOI was 10 μM.

[0594] 4) Add 1 μL DMSO to the high signal control wells (High control) and 1 μL DMSO to the low signal control wells (Low

[0595] control)5-HT 1A 1 μL of 2 mM 8-OH-DPAT (final concentration 10 μM) was added to the plate, 5-

[0596] HT 2A Add 1 μL of 200 μM Ketanserin (final concentration 1000 nM) and 5-HT 2B1 μL of 10 mM Serotonin was added to the plate (final concentration 50 μM).

[0597] 5) Add 100 μL of the corresponding cell membrane to each well.

[0598] 6) Add 100 μL of the corresponding isotope to each well.

[0599] 7) The 96-well plate was sealed and placed on a shaker at room temperature and 300 rpm for 1 hour. Meanwhile, the GF / C filter plate was soaked with 0.3% PEI.

[0600] 8) After incubation, the cells were collected onto a GF / C filter plate using a cell collector, washed 4 times with plate washing buffer, and dried in an oven at 50°C for 1 hour.

[0601] 9) Seal the bottom of the dried GF / C filter plate with a film, add 50 μL of scintillation fluid to each well, and seal. Read using Microbeta.

[0602] Table 4. Detection buffers used for different targets

[0603] Target Assay Buffer <![CDATA[5-HT 1A ]]> <![CDATA[50mM Tris-HCl pH 7.4;10mM MgSO4;0.5mM EDTA]]> <![CDATA[5-HT 2A ]]> 50mM Tris-HCl pH7.4 <![CDATA[5-HT 2B ]]> <![CDATA[50mM Tris-HCl pH 7.4;4mM CaCl2;0.1%ascorbic acid]]>

[0604] 4. Data Processing and Analysis

[0605] 1) Calculate the activity percentage using Microsoft Excel software, formula: %Inhibition = 100 × (1-

[0606] (Sample Raw Value-Low Control Average) / (High Control Average-LowControl Average))

[0607] 2) The inhibition rate was fitted using the four-parameter equation “log (inhibitor) vs response-variable slope” in GraphPad Prism 5.0 to calculate the IC 50 .

[0608] 3) Calculate K i Value, formula: K i =IC 50 / (1+[L]Kd)

[0609] 5. Test results

[0610] Compound b, compound c and compound e have an effect on 5-hydroxytryptamine receptor (5-HT 1A , 5-HT 2A and 5-HT 2B ) Affinity Ki As shown in Table 5.

[0611] Table 5. Affinity of Compound B, Compound C, and Compound E for each target

[0612]

[0613] 6. Conclusion

[0614] Compound b and compound c have an effect on 5-HT 1A , 5-HT 2A and 5-HT 2B The affinity of compound e for 5-HT receptor is stronger than that of compound e for 5-HT 1A , 5-HT 2A and 5-HT 2B Receptor affinity.

[0615] Biological Example 2 Cell Functional Test

[0616] Compounds for D2 and 5-HT 1A Cellular functional assay of receptors.

[0617] 1. Purpose of the experiment

[0618] Using FLIPR calcium flux detection technology, the effects of compounds on D2 receptors and 5-HT were detected at the cellular level. 1A Receptor stimuli.

[0619] 2. Experimental Materials and Instruments

[0620] 2.1 Test products

[0621] Preparation method: Dissolve the test compounds in DMSO to make 10mM stock solution and store in a 4℃ refrigerator.

[0622] 2.2 Cell lines

[0623] Stable expression of D2 and 5-HT 1A The cell line information of the receptor is as follows in Table 6:

[0624] Table 6. Cell line information

[0625]

[0626] 2.3 Buffer

[0627] 1) Assay buffer: HBSS buffer containing 2 mM HEPES.

[0628] 2) Probenecid (250 mM) solution: press Fluo-4 Direct TMAccording to the kit instructions, add 77 mg probenecid to 1 ml test buffer and prepare it before use.

[0629] 3) Fluo-4 Direct TM (2X, 8M) loading buffer: According to the kit instructions, add the required amount of Fluo-4 Direct TM Take out and melt, add 10mL test buffer and 0.2mL 250mM Probenecid solution to each tube, vortex and shake in the dark for more than 5 minutes, and prepare before use.

[0630] 2.4 Reagents and consumables

[0631] Table 7. Reagents and consumables

[0632] name Manufacturer DMEM medium Invitrogen F12 medium Invitrogen L-Glutamate Invitrogen Fetal bovine serum Hyclone G418 Invitrogen Hygromycin Invitrogen Penicillin-Streptomycin Hyclone HBSS Invitrogen HEPES Invitrogen <![CDATA[Fluo-4Direct TM Kit]]> Invitrogen 384-well poly-lysine coated cell plates Greiner 384-well compound plate Greiner Labcyte PP 384plate Labcyte (±)-SKF-38393 Sigma Dopamine Sigma 5-CT Sigma SCH-23390 Sigma Amisulpride Sigma Methiothepin mesylate Sigma 5-HT Sigma

[0633] 2.5 Instruments and Equipment

[0634] Table 8. Instruments and equipment

[0635] instrument model Manufacturer FLIPR FT-0249 Molecular Device Bravo USS2009180003 Agilent Technologies Echo ECHO 555 Labcyte Cell Counter AN16097 Beckman

[0636] 3. Experimental Methods

[0637] Day 1: Cell plating

[0638] 1) For frozen cells, remove the cells from liquid nitrogen and quickly place them in a 37°C water bath. Add 10 mL of preheated culture medium to suspend the cells, remove 1 mL and use Vi-CELL TM Count and measure the activity; for cultured cells, wash twice with preheated DPBS, then add appropriate amount of trypsin to digest at 37℃ for 1 minute; then add appropriate amount of culture medium to terminate digestion, take out 1mL and use Vi-CELL TM Count and determine viability.

[0639] 2) Centrifuge the cells at 1000 rpm for 5 minutes.

[0640] 3) Gently discard the supernatant without touching the cell mass at the bottom.

[0641] 4) Dilute the cells to 1×10 6 Cells / mL (20,000 cells / 20 μL) were seeded in 384-well poly-lysine-coated cell plates and cultured at 37° C., 5% CO 2 for 16-20 hours.

[0642] Day 2: Sample addition and detection

[0643] 1)EC 80 Detection:

[0644] a) To EC80 2×Fluo-4 detection reagent was added to the detection cell plate, 20 μL per well, incubated in a 37°C incubator for 50 min, and then allowed to stand at room temperature for 10 min.

[0645] b) The compound was diluted 3-fold with DMSO to 10 concentrations using Echo 555 and added to the EC 80 In the test compound plate 1, 500 nL was added to each well in duplicate, and then 30 μL of the detection buffer was added to each well.

[0646] Table 9. Starting concentrations of positive compounds for each target agonist

[0647] Cell Name Agonists Starting concentration <![CDATA[D1]]> (±)-SKF-38393 360nM <![CDATA[D2]]> Dopamine 180nM <![CDATA[5-HT 1A ]]> 5-CT 180nM <![CDATA[5-HT 2A ]]> 5-CT 300nM <![CDATA[5-HT 2B ]]> 5-HT 600nM

[0648] c) EC 80 Detection cell plate, EC 80 Test compound plate, place pipette tips into FLIPR instrument, start the instrument, transfer 10 μL reference compound from compound plate 1 to the cell plate, read, and calculate EC 80 .

[0649] 2) Compound activity detection

[0650] a) Add 2×Fluo-4 detection reagent to each well of the cell plate, incubate in a 37°C incubator for 50 min, and then let stand at room temperature for 10 min.

[0651] b) The compound was diluted 4-fold with DMSO into 10 concentrations by Echo 555 and added to compound plate 2, 900 nL per well, duplicate wells, and then 30 μL of detection buffer was added to each well. The starting concentration of the test compound for agonist detection was 12 μM, and the starting concentration for antagonist detection was 10

[0652] μM.

[0653] Table 10. Starting concentrations of positive antagonist compounds for each target

[0654] Cell Name Agonists Starting concentration Antagonists Starting concentration <![CDATA[D1]]> (±)-SKF-38393 360nM SCH-23390 75nM <![CDATA[D2]]> Dopamine 180nM Amisulpride 250nM <![CDATA[5-HT 1A ]]> 5-CT 180nM Methiothepin mesylate 7500nM <![CDATA[5-HT 2A ]]> 5-CT 300nM Methiothepin mesylate 2500nM <![CDATA[5-HT 2B ]]> 5-HT 600nM Methiothepin mesylate 10000nM

[0655] c) Prepare 6*EC of the compounds respectively 80 And added to compound plate 3, 30 μL per well.

[0656] d) Place the cell plate, the corresponding compound plate 2, and the pipette tip into the FLIPR instrument, start the instrument, transfer 10 μL of compound from compound plate 2 to the cell plate, read the number, and calculate the

[0657] EC 50 ;

[0658] e) Take out compound plate 2 and put it into the corresponding EC 80 Compound plate 3, transfer 10 μL of compound from compound plate 3 to the cell plate, read, and calculate the IC of each compound 50 .

[0659] 4. Data Processing and Analysis

[0660] 1) Calculate the activity percentage using Microsoft Excel software, using the formula %Effect =

[0661] 100×(Sample Raw Value-Low Control Average) / (High Control Average-

[0662] Low Control Average); for inhibitors, the formula %Inhibition=100×(1-(Sample

[0663] Raw Value-Low Control Average) / (High Control Average–HLow Control

[0664] Average)

[0665] 2) Using GraphPad Prism 5 data analysis software, log[agonist] vs.

[0666] The response--Variable slope mode is used for fitting analysis. For the inhibitor, log[inhibitior] is used.

[0667] vs.response--Variable slope mode is used for fitting analysis to obtain the

[0668] IC 50 / EC 50 value.

[0669] 5. Test results

[0670] This experiment detected the agonist and antagonist effects of compound b, compound c and compound e on multiple receptors at the cellular level. The results are shown in Tables 11 and 12.

[0671] Table 11. Effects of Compound b, Compound c, and Compound e on D2 and 5-HT 1A Receptor agonism

[0672]

[0673] Table 12. Effects of Compound b and Compound c on D1, D2, and 5-HT 1A , 5-HT 2A , 5-HT 2B Receptor antagonism

[0674]

[0675] 6. Conclusion

[0676] The test results showed that compound e had an effect on 5-HT 1A Compounds b and c did not show agonist effects on 5-HT receptors. 1A The receptors showed agonist effects. Compound b and compound c had agonist effects on D1, D2, and 5-HT 1A , 5-HT 2A and 5-HT 2B The receptors showed a certain antagonistic effect.

[0677] Biological Example 3 Cellular functional assay of the test compound on D3 receptor.

[0678] 1. Purpose of the experiment

[0679] Evaluation of the biological activity of the compounds.

[0680] 2. Experimental Methods

[0681] The test method (dopamine D3, GTPγS binding) is as follows:

[0682]

[0683] 3. Experimental results

[0684] The test data are shown in Table 13 below:

[0685] Table 13. Antagonistic effects of compound b, compound c, and compound e on D3 receptor

[0686] Compound Detection concentration Inhibition percentage Compound b 1μM 105％ Compound c 1μM 100％ Compound e 1μM 74％

[0687] 4. Conclusion

[0688] The test results showed that the antagonistic effect of compound b and compound c on D3 receptor was greater than that of compound e on D3 receptor.

[0689] Biological Example 4 hERG Toxicity Study

[0690] 1. Purpose of the experiment

[0691] The rapidly activated delayed rectifier potassium channel (IKr) encoded by hERG (human Ether-à-go-go-Related Gene) is an important ion channel involved in the formation of phase 3 repolarization of the cardiac action potential. Drug blockade of the hERG channel can lead to prolonged cardiac repolarization, which is manifested as a prolonged QT interval on the electrocardiogram, known as long QT syndrome. In some cases, drug-induced delayed ventricular repolarization may cause fatal torsade de pointes (TdP) arrhythmia.

[0692] This experiment used manual patch clamp technology (the gold standard for hERG safety evaluation) to study the inhibitory effect of the test compound on hERG potassium channels and evaluate its risk of inducing ventricular repolarization toxicity.

[0693] 2. Experimental materials and instruments:

[0694] 2.1 Reagents

[0695] Table 14. Chemical reagent information

[0696]

[0697] 2.2 Instruments

[0698] Table 15. Instrument information

[0699]

[0700]

[0701] 3. Experimental steps:

[0702] 3.1 Cell culture

[0703] In this experiment, HEK-293 cell lines stably expressing hERG potassium channel were used. hERG potassium channel cells were purchased from Creacell (Cat. No.: A-0320). The cell culture method is as follows:

[0704] The HEK-293 cell line stably expressing hERG potassium channel was cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg / mL G418 at a culture temperature of 37° C. and a carbon dioxide concentration of 5%.

[0705] Cell passaging: Remove old culture medium and wash once with PBS, then add 1 mL of TrypLE TM

[0706] Express solution, incubate at 37℃ for about 0.5min. When the cells detach from the bottom of the dish, add about 5mL of complete medium preheated at 37℃. Use a pipette to gently blow the cell suspension to separate the aggregated cells. Transfer the cell suspension to a sterile centrifuge tube and centrifuge at 1000rpm for 5min to collect the cells. For expansion or maintenance culture, inoculate the cells in a 6cm cell culture dish, with 2.5×10 cells in each cell culture dish. 5 cells (final volume: 5 mL).

[0707] To maintain the electrophysiological activity of cells, the cell density must not exceed 80%.

[0708] Before patch clamping, cells were stained with TrypLE TM Express separation, 4×10 3 The cells were plated on coverslips and cultured in 24-well plates (final volume: 500 μL). After 18 hours, the assay was performed.

[0709] 3.2 Electrophysiological recording

[0710] 3.2.1 Record the liquid used

[0711] Extracellular fluid:

[0712] 140mM NaCl, 3.5mM KCl, 1mM MgCl2·6H2O, 2mM CaCl2·2H2O, 10mM D-Glucose, 10mM HEPES, 1.25mM NaH2PO4·2H2O, pH adjusted to 7.4 with NaOH.

[0713] Intracellular fluid:

[0714] 20mM KCl, 115mM K-Aspartic, 1mM MgCl2·6H2O, 5mM EGTA, 10mM HEPES, 2mM Na2-ATP, KOH adjust pH=7.2.

[0715] The extracellular solution can be stored for 2 weeks. After the intracellular solution is prepared, it is divided into 1 mL per tube and stored in a -20℃ refrigerator. Freshly thawed intracellular solution is used for daily testing. All intracellular solutions should be used up within three months. If it exceeds three months, the old intracellular solution should be discarded and re-prepared.

[0716] 3.2.2 Patch clamp assay

[0717] The voltage stimulation scheme for recording hERG current by whole-cell patch clamp is as follows: After the whole-cell seal is formed, the cell membrane voltage is clamped at -80mV. The clamping voltage is depolarized from -80mV to -50mV for 0.5s (as a leakage current detection), then stepped to 30mV for 2.5s, and then quickly restored to -50mV for 4s to stimulate the tail current of the hERG channel. Data is collected repeatedly every 10s to observe the effect of drugs on the hERG tail current. The experimental data are collected by the IPA amplifier (Sutter Instrument) and stored in the SutterPatch (with Igor Pro) software.

[0718] The patch clamp operation first uses a microelectrode puller to pull a capillary glass tube into a recording electrode, then loads the electrode filled with intracellular fluid into the microelectrode holder, and manipulates the microelectrode manipulator under an inverted microscope to immerse the electrode in the extracellular fluid and record the electrode resistance (Rpip). Then slowly contact the electrode to the cell surface, apply negative pressure to form a GΩ seal. At this time, fast capacitance compensation is performed, and negative pressure is continued to be applied to break the cell membrane to form a whole-cell recording mode. Finally, slow capacitance compensation is performed and experimental parameters such as series resistance (Rs) are recorded. No leakage compensation is given.

[0719] When the hERG current recorded by the whole cell is stable, the drug is administered. Each drug concentration acts for about 5 minutes (or the current stabilizes) before the next concentration is detected. Multiple concentrations are detected for each test compound. The coverslip with cells is placed in a recording bath under an inverted microscope. The blank control external solution and the working solution of the test compound flow through the recording bath from low concentration to high concentration in sequence by gravity perfusion to act on the cells. A peristaltic pump is used for liquid exchange during recording. The current detected by each cell in the external solution without the compound serves as its own control group. At least three cells are used for each concentration and the test is repeated three times independently. All electrophysiological tests are performed at room temperature.

[0720] 4. Data Analysis:

[0721] First, the peak tail current after each drug concentration is compound ) and blank control tail current (Peak tail current control ) and then calculate the inhibition rate corresponding to each drug concentration The mean (Mean), standard deviation (SD) and standard error (SE) of the inhibition rate at each concentration were calculated, and the data were expressed as Mean±SE.

[0722] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0723] The IC of each compound was calculated using the above equation. 50 The dose-effect curve was fitted nonlinearly, where IC 50 is the half inhibitory concentration. IC 50 The calculation and curve fitting were completed using GraphPad Prism software.

[0724] 5. Experimental Results

[0725] The inhibitory effects of the test compounds b and d on hERG channels were detected in three independent repeated experiments, and the IC values ​​of the test compounds on hERG channel current were calculated by fitting the concentration-effect curves. 50 value.

[0726] Table 16. IC of Compound b and Compound d on hERG channel current 50 value

[0727] Compound <![CDATA[IC 50 ]]> Compound b 3.401μM Compound d 1.585μM

[0728] The test results show that the inhibitory effect of compound b on the hERG channel is lower than that of compound d, showing better safety.

[0729] Synthesis Example 1

[0730]

[0731] In a 50 mL three-necked flask, compound 1 (0.050 g, 0.117 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (21 mg).

[0732] Synthesis Example 2

[0733]

[0734] In a 50 mL three-necked flask, compound 2 (0.050 g, 0.112 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (19 mg).

[0735] Synthesis Example 3

[0736]

[0737] In a 50 mL three-necked flask, compound 3 (0.050 g, 0.113 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (26 mg).

[0738] Synthesis Example 4

[0739]

[0740] In a 50 mL three-necked flask, compound 4 (0.050 g, 0.109 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (31 mg).

[0741] Synthesis Example 5

[0742]

[0743] In a 50 mL three-necked flask, compound 5 (0.050 g, 0.113 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (30 mg).

[0744] Synthesis Example 6

[0745]

[0746] In a 50 mL three-necked flask, compound 6 (0.050 g, 0.109 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇=1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (24 mg).

[0747] Synthesis Example 7

[0748]

[0749] In a 50 mL three-necked flask, compound 7 (0.050 g, 0.109 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (15 mg).

[0750] Synthesis Example 8

[0751]

[0752] In a 50 mL three-necked flask, compound 8 (0.050 g, 0.106 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (29 mg).

[0753] Synthesis Example 9

[0754]

[0755] In a 50 mL three-necked flask, compound 9 (0.050 g, 0.106 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (26 mg).

[0756] Synthesis Example 10

[0757]

[0758] In a 50 mL three-necked flask, compound 10 (0.050 g, 0.099 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (18 mg).

[0759] Synthesis Example 11

[0760]

[0761] In a 50 mL three-necked flask, compound 10 (0.050 g, 0.099 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (28 mg).

[0762] Synthesis Example 12

[0763]

[0764] In a 50 mL three-necked flask, compound 12 (0.050 g, 0.100 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (24 mg).

[0765] Synthesis Example 13

[0766]

[0767] In a 50 mL three-necked flask, compound 14 (0.050 g, 0.056 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (17 mg).

[0768] Synthesis Example 14

[0769]

[0770] In a 50 mL three-necked flask, compound 17 (0.050 g, 0.064 mmol) was dissolved in methanol (2 mL), and 2 mL of 1 M HCl was added. The mixture was stirred at room temperature for 0.5 h. After the reaction was completed, an appropriate amount of Na2CO3 aqueous solution was added to adjust the pH to 8.0. The solvent was evaporated and silica gel was added to mix the sample. The mixture was then purified by column chromatography (DCM / MeOH=20 / 1) to obtain compound b (32 mg).

[0771] Synthesis Example 15

[0772]

[0773] In a 50 mL three-necked flask, compound 18 (0.050 g, 0.112 mmol) was dissolved in methanol (2 mL), and 2 mL of 1 M HCl was added. The mixture was stirred at room temperature for 0.5 h. After the reaction was completed, an appropriate amount of Na2CO3 aqueous solution was added to adjust the pH to 8.0. The solvent was evaporated and silica gel was added to mix the sample. The mixture was then purified by column chromatography (DCM / MeOH=20 / 1) to obtain compound b (13 mg).

[0774] Synthesis Example 16

[0775]

[0776] In a 50 mL three-necked flask, compound 19 (0.050 g, 0.112 mmol) was dissolved in methanol (2 mL), and 2 mL of 1 M HCl was added. The mixture was stirred at room temperature for 0.5 h. After the reaction was completed, an appropriate amount of Na2CO3 aqueous solution was added to adjust the pH to 8.0. The solvent was evaporated and silica gel was added to mix the sample. The mixture was then purified by column chromatography (DCM / MeOH=20 / 1) to obtain compound b (21 mg).

[0777] Synthesis Example 17

[0778]

[0779] In a 50 mL three-necked flask, compound 20 (0.050 g, 0.120 mmol) was dissolved in methanol (2 mL), 1 M HCl 2 mL was added, and the mixture was stirred at room temperature for 0.5 h. After the reaction was completed, an appropriate amount of Na2CO3 aqueous solution was added to adjust the pH to 8.0. The solvent was evaporated and silica gel was added to mix the sample. The mixture was then purified by column chromatography (DCM / MeOH=20 / 1) to obtain compound b (15 mg).

[0780] Synthesis Example 18

[0781]

[0782] In a 50 mL three-necked flask, compound 21 (0.050 g, 0.120 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (18 mg).

[0783] Synthesis Example 19

[0784]

[0785] In a 50 mL three-necked flask, compound 23 (0.050 g, 0.097 mmol) was dissolved in 2 mL of a mixed solvent (V 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (21 mg).

[0786] Synthesis Example 20

[0787]

[0788] In a 50 mL three-necked flask, compound 24 (0.050 g, 0.094 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (22 mg).

[0789] Synthesis Example 21

[0790]

[0791] In a 50 mL three-necked flask, compound 25 (0.050 g, 0.097 mmol) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), 2 mL of 20% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hour. After the reaction was completed, 2 mL of water was added, and the mixture was stirred for another 0.5 hour to precipitate a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain compound b (25 mg).

[0792] Synthesis Example 22

[0793]

[0794] In a 50 mL three-necked flask, compound 26 (0.050 g, 0.095 mmol, 1.0 eq) was dissolved in 2 mL of a mixed solvent (V) consisting of tetrahydrofuran and methanol. 四氢呋喃 :V 甲醇 =1:1), add 2 mL of 20% sodium hydroxide aqueous solution, stir at room temperature for 0.5 hour, add 2 mL of water after the reaction is completed, stir for another 0.5 hour, precipitate solid, filter and wash the filter cake with water, and dry to obtain compound b (30 mg).

Claims

1. A method for preparing trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazin-1-yl)ethyl)cyclohexyl)urea, comprising the step of hydrolyzing a cyclohexane derivative of formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof to obtain trans-4-(2-(4-(benzo[b]thiophen-7-yl)piperazin-1-yl)ethyl)cyclohexyl)urea: in, R1, R2 and R3 are each independently selected from H or a leaving group, and at least one of R1, R2 and R3 is a leaving group; Optionally, any two of said R1, R2 and R3 may be connected to form a ring; The hydrolysis is acid hydrolysis or alkaline hydrolysis; In the acid hydrolysis, the acid is preferably an inorganic acid or an organic acid; The inorganic acid is preferably hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid or phosphoric acid, more preferably hydrochloric acid or sulfuric acid, further preferably hydrochloric acid; The organic acid is preferably oxalic acid, maleic acid, acetic acid, formic acid or tartaric acid; In the alkaline hydrolysis, the base is preferably an inorganic base, and the inorganic base is preferably sodium hydroxide, potassium hydroxide or sodium hydrogen; Optionally, the molar ratio of the acid / base to the cyclohexane derivative is 1:100 to 100:1, preferably 10-20:

1.

2. The preparation method according to claim 1, wherein the structure of the cyclohexane derivative is as follows: in, R1 is a leaving group; or, The structure of the cyclohexane derivative is as follows: Wherein, R3 is a leaving group; or, The structure of the cyclohexane derivative is as follows: Wherein, R1 and R2 are each independently selected from a leaving group, and optionally, R1 and R2 can be connected to form a ring; or, The structure of the cyclohexane derivative is as follows: Wherein, R1 and R3 are each independently selected from a leaving group, and optionally, R1 and R3 can be connected to form a ring; or, The structure of the cyclohexane derivative is as follows: Said R1, R2 and R3 are each independently selected from a leaving group, and optionally, R1 and R2 may be connected to form a ring, and / or R1 and R3 may be connected to form a ring, and / or R2 and R3 may be connected to form a ring; The rings are preferably connected to form a five-membered, six-membered, seven-membered or eight-membered heterocyclic ring.

3. The preparation method according to any one of claims 1 to 2, wherein the leaving group is selected from hydroxyl, sulfhydryl, alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy, haloalkylthio, aldehyde, ester, thioester, acyl, ester acyl, amide, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, cycloalkyl, cycloalkyloxy, cycloalkylthio, heterocyclyl, heterocyclyloxy, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy or heteroarylthio, preferably alkyl, alkoxy, haloalkyl, haloalkoxy, aldehyde, ester, acyl, ester acyl, amide, cycloalkyl, cycloalkyloxy, heterocyclyl, heterocyclyloxy, aryloxy, heteroaryloxy; optionally, it can be further substituted.

4. The preparation method according to claim 1, wherein the leaving group is in, m is an integer from 0 to 10, specifically 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; n is an integer from 1 to 10, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; R4, R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it can be further substituted; R6 is each independently selected from hydrogen, sulfonyl, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R7 is each independently selected from hydrogen, hydroxy, thiol, sulfonic acid, sulfonyl, phosphate, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R8 is each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sugar group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, acyl, acylamino, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; Optionally, any two of R4, R5, R6, R7 and R8 may be connected to form a ring.

5. The preparation method according to claim 1, wherein the leaving group is or, The leaving group is in, n is an integer from 1 to 10, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; X1 and X2 are each independently selected from -O-, -S- or -NR7-; R4, R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it can be further substituted; R6 is each independently selected from hydrogen, sulfonyl, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R7 is each independently selected from hydrogen, hydroxy, thiol, sulfonic acid, sulfonyl, phosphate, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R8 is each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sugar group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, acyl, acylamino, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R9 is selected from R 10 Selected from -O-, Optionally, any two of R4, R5, R6, R7 and R8 may be connected to form a ring.

6. According to the preparation method according to any one of claims 1 to 5, the leaving group is 7. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, m, R4, R5 and R6 are as defined above.

8. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, n, R4, R5, and R7 are as defined above.

9. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, n, R4, R5, R7, and R8 are as defined above.

10. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, m, R4, R5 and R6 are as defined above.

11. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, R8 is defined as above.

12. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, m, R4, R5, and R7 are as defined above.

13. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, m, R4, R5 and R6 are as defined above.

14. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, n, X1, X2, R4, R5, and R8 are as defined above.

15. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, R4, R5, and R8 are as defined above.

16. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, n, X1, R4, R5, and R8 are as defined above.

17. The preparation method according to claim 4, wherein the structure of the cyclohexane derivative is as follows: in, n, X1, R4, R5 and R6 are as defined above.

18. The preparation method according to claim 1, wherein the structure of the cyclohexane derivative is as follows: in, R 11 is a removable linking group; g is selected from 2, 3 or 4.

19. The preparation method according to claim 18, wherein the structure of the cyclohexane derivative is as follows: in, R9 is selected from R 10 Selected from -O-, n, X1, X2, R4 and R5 are as defined above.

20. The preparation method according to any one of claims 1 to 19, wherein the structure of the cyclohexane derivative is as follows:

21. A cyclohexane derivative, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the structure of the cyclohexane derivative is as follows: in, R1, R2 and R3 are each independently selected from H or a leaving group, and at least one of R1, R2 and R3 is a leaving group; Optionally, any two of said R1, R2 and R3 may be connected to form a ring; The cyclohexane derivatives do not include the following compounds:

22. The cyclohexane derivative, its stereoisomer or a pharmaceutically acceptable salt thereof according to claim 21, wherein the structure of the cyclohexane derivative is as follows: in, R1 is a leaving group; or, The structure of the cyclohexane derivative is as follows: Wherein, R3 is a leaving group; or, The structure of the cyclohexane derivative is as follows: Wherein, R1 and R2 are each independently selected from a leaving group, and optionally, R1 and R2 can be connected to form a ring; or, The structure of the cyclohexane derivative is as follows: Wherein, R1 and R3 are each independently selected from a leaving group, and optionally, R1 and R3 can be connected to form a ring; or, The structure of the cyclohexane derivative is as follows: Said R1, R2 and R3 are each independently selected from a leaving group, and optionally, R1 and R2 may be connected to form a ring, and / or R1 and R3 may be connected to form a ring, and / or R2 and R3 may be connected to form a ring; The rings are preferably connected to form a five-membered, six-membered, seven-membered or eight-membered heterocyclic ring.

23. The cyclohexane derivative according to any one of claims 21 to 22, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the leaving group is selected from hydroxyl, thiol, alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy, haloalkylthio, aldehyde, ester, thioester, acyl, esteracyl, amide, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, cycloalkyl, cycloalkyloxy, cycloalkylthio, heterocyclyl, heterocyclyloxy, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy or heteroarylthio, preferably alkyl, alkoxy, haloalkyl, haloalkoxy, aldehyde, ester, acyl, esteracyl, amide, cycloalkyl, cycloalkyloxy, heterocyclyl, heterocyclyloxy, aryloxy, heteroaryloxy; optionally, it can be further substituted.

24. The cyclohexane derivative according to claim 23, its stereoisomer or pharmaceutically acceptable salt thereof, wherein the hydroxyl, thiol, alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy, haloalkylthio, aldehyde, ester, thioester, acyl, ester acyl, amide, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, cycloalkyl, cycloalkyloxy, cycloalkylthio, heterocyclyl, heterocyclyloxy, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy or heteroarylthio is further substituted with one or more of the following groups: hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic group, aryl or heteroaryl; optionally, it can be further substituted, for example, further substituted by one or more of hydrogen, halogen, nitro, hydroxyl, sulfhydryl, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclic group, aryl or heteroaryl.

25. The cyclohexane derivative according to any one of claims 21 to 24, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the leaving group is selected from a hydroxyl group, a thiol group, a C1-20 alkyl group, a C1-20 alkoxy group, a C1-20 alkylthio group, a C1-20 haloalkyl group, a C1-20 haloalkoxy group, a C1-20 haloalkylthio group, an aldehyde group, a C1-20 ester group, a C1-20 thioester group, a C1-20 acyl group, a C1-20 ester acyl group, a C1-20 amide group, a sulfonic acid group, a sulfonyl group, a phosphate group, a phosphoryl group, a D-(+)-pyranose glucopyranose group, a D-(+)-pyranose glucopyranose ester group, a 3-20 membered cycloalkyl group, a 3-20 membered cycloalkyloxy group, a 3-20 membered cycloalkylthio group, a 3-2 0-membered heterocyclyl, 3-20-membered heterocyclyloxy, 3-20-membered heterocyclylthio, 6-20-membered aryl, 6-20-membered aryloxy, 6-20-membered arylthio, 5-20-membered heteroaryl, 5-20-membered heteroaryloxy or 5-20-membered heteroarylthio, preferably C1-20 alkyl, C1-20 alkoxy, C1-20 haloalkyl, C1-20 haloalkoxy, C1-20 aldehyde, C1-20 ester, C1-20 acyl, C1-20 ester acyl, C1-20 amido, 3-20-membered cycloalkyl, 3-20-membered cycloalkyloxy, 3-20-membered heterocyclyl, 3-20-membered heterocyclyloxy, 6-20-membered aryloxy, 6-20-membered heteroaryloxy; optionally, it may be further substituted.

26. The cyclohexane derivative according to any one of claims 21 to 25, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the structural unit Selected from Preferably 27. The cyclohexane derivative, stereoisomer thereof or pharmaceutically acceptable salt thereof according to claim 21, wherein the leaving group is in, m is an integer from 0 to 10, specifically 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; n is an integer from 1 to 10, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; R4, R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it can be further substituted; R6 is each independently selected from hydrogen, sulfonyl, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R7 is each independently selected from hydrogen, hydroxy, thiol, sulfonic acid, sulfonyl, phosphate, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R8 is each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sugar group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, acyl, acylamino, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; Optionally, any two of R4, R5, R6, R7 and R8 may be connected to form a ring.

28. The cyclohexane derivative according to claim 21, its stereoisomer or a pharmaceutically acceptable salt thereof, The leaving group is or, The leaving group is Best in, n is an integer from 1 to 10, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; X1 and X2 are each independently selected from -O-, -S- or -NR7-; R4, R5 are each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sulfonic acid, sulfonyl, phosphate, phosphoryl, sugar group, sugar ester group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, haloalkoxy, acyl, acylamino, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it can be further substituted; R6 is each independently selected from hydrogen, sulfonyl, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R7 is each independently selected from hydrogen, hydroxy, thiol, sulfonic acid, sulfonyl, phosphate, phosphoryl, alkyl, haloalkyl, hydroxyalkyl, acyl, thioester, ester, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R8 is each independently selected from hydrogen, halogen, nitro, hydroxyl, thiol, cyano, amino, amine, sugar group, amino acid group, alkyl, haloalkyl, hydroxyalkyl, acyl, acylamino, cycloalkyl, heterocyclyl, aryl or heteroaryl; optionally, it may be further substituted; R9 is selected from R 10 Selected from -O-, Optionally, any two of R4, R5, R6, R7 and R8 may be connected to form a ring.

29. The cyclohexane derivative according to any one of claims 21 to 28, its stereoisomer or a pharmaceutically acceptable salt thereof, The leaving group is 30. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, m, R4, R5 and R6 are as defined above.

31. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, n, R4, R5, and R7 are as defined above.

32. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, n, R4, R5, R7, and R8 are as defined above.

33. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, m, R4, R5 and R6 are as defined above.

34. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, R8 is defined as above.

35. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, m, R4, R5, and R7 are as defined above.

36. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, m, R4, R5 and R6 are as defined above.

37. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, n, X1, X2, R4, R5, and R8 are as defined above.

38. The cyclohexane derivative according to claim 37, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, R4, R5, and R8 are as defined above.

39. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, n, X1, R4, R5, and R8 are as defined above.

40. The cyclohexane derivative according to any one of claims 28 to 29, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, n, X1, R4, R5 and R6 are as defined above.

41. The cyclohexane derivative according to claim 21, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, R 11 is a removable linking group; g is selected from 2, 3 or 4.

42. The cyclohexane derivative according to claim 41, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows: in, R9 is selected from R 10 Selected from -O-, n, X1, X2, R4 and R5 are as defined above.

43. The cyclohexane derivative according to any one of claims 21 to 42, its stereoisomer or a pharmaceutically acceptable salt thereof, The structure of the cyclohexane derivative is as follows:

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