Dihydropyrazole azepine compound, pharmaceutical composition containing the same and application thereof in anti-tumor

Abstract

The present invention discloses a dihydropyrazole-azepine compound, a pharmaceutical composition containing the same, and its use in anti-tumor treatment. The structure of the dihydropyrazole-azepine compound is shown in Formula I. The inventors of the present invention have confirmed through multiple experiments that the compound of the present invention has a significant inhibitory effect on AKT1, exhibiting a potent anti-proliferative effect on tumor cell lines such as mouse neuroblastoma Neuro2a cells and a potent differentiation-inducing effect on Neuro2a cell lines. Therefore, the compound of the present invention can be used as a differentiation-inducing regulator and / or AKT inhibitor in drugs for treating solid tumors or blood cancers associated with cell proliferation and differentiation abnormalities in humans or animals.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and specifically designs dihydropyrazole azepine compounds as AKT inhibitors, pharmaceutical compositions containing the same, and their use in inducing differentiation and anti-tumor. Background Art

[0002] Akt, or protein kinase B, has attracted considerable attention as a promising anti-tumor target. Akt is a core component of the PI3K / Akt signaling pathway. The Akt family comprises three isoforms: Akt1, Akt2, and Akt3, sharing over 80% sequence homology. Studies have shown that the different Akt isoforms are highly identical in structure and function, though their expression levels vary in different tumors. Akt can directly phosphorylate mTOR, Bad, and Caspase9 proteins, and regulate the ForkHead family of transcription factors and NF-κB, thereby controlling numerous cellular processes crucial for tumor development and progression, including transcription, translation, metabolism, apoptosis, and angiogenesis. Studies have also shown that Akt is overexpressed or dysregulated in many tumors. Akt abnormalities are closely associated with the development and progression of these tumors, as well as resistance to chemotherapy and radiotherapy. Both in vitro and in vivo pharmacological studies have demonstrated that Akt inhibitors can promote programmed cell death in cancer cells.

[0003] Abnormal tumor cell differentiation is a key process in tumor development and progression. Induction of differentiation refers to a therapeutic approach in which malignant tumor cells, under the influence of differentiation inducers in vitro or in vivo, reverse their differentiation to normal or near-normal cell types, thereby reversing the malignant phenotype. The discovery of effective differentiation inducers will transform the previous treatment paradigm, which primarily targets tumor cells for killing, toward a new approach focused on controlling and modulating their biological behavior, ultimately achieving a cure for tumors. Over the past 20 years, over a hundred compounds with differentiation effects have been discovered, a significant number of which have entered Phase I and II clinical trials. A few, such as retinoids (RA), arsenic compounds, interferon-γ (IFN-γ), and nerve growth factor (NGF), have been used as differentiation inducers in clinical tumor treatment and prevention, demonstrating promising efficacy. Therefore, the research and application of differentiation inducers has opened up a broad avenue for the study of oncology drugs and their pharmacology. In recent years, the therapeutic role of targeted drugs in tumor differentiation has become increasingly prominent. Akt occupies a key node in the tumor differentiation-related signaling pathway, and its inhibitors may have the potential to induce tumor cell differentiation.

[0004] In summary, using AKT inhibitors to develop small molecule drugs that have both differentiation-inducing therapeutic effects and proliferation-inhibiting effects is expected to provide clinical drugs with a new mechanism of action for tumor treatment. Summary of the Invention

[0005] The purpose of the present invention is to provide a novel dihydropyrazole azepine compound having strong anticancer effect, strong differentiation inducing effect and AKT inhibitory effect, and its optical isomers or pharmaceutically acceptable salts or solvates.

[0006] Terminology: As used herein, the term "aryl" refers to an all-carbon monocyclic or fused polycyclic group of 5 to 12 carbon atoms with a completely conjugated electron system. Non-limiting examples of aromatic rings include benzene, naphthalene, and anthracene. Aromatic rings may be unsubstituted or substituted. Substituents on aromatic rings are selected from halogen, nitro, amino, cyano, hydroxyl, C l ~C6 alkyl, C l ~C6 alkoxy, halogenated C l ~C6 alkyl, halogenated C l ~C6 alkoxy, C3~C6 cycloalkyl, halogenated C3~C6 cycloalkyl.

[0007] The term "heteroaromatic" as used herein refers to an unsaturated carbon ring of 5 to 12 carbon atoms with a completely conjugated electron system, which is equivalent to one or more carbons in the above-mentioned "aryl" being replaced by heteroatoms such as oxygen, nitrogen, sulfur, etc. The heteroaromatic ring can be a monocyclic ring or a bicyclic ring, that is, it is formed by the fusion of two rings. Specific heteroaromatic groups can be: pyridyl, pyrimidinyl, pyrazinyl, isoxazolyl, isothiazolyl, pyrazolyl, thiazolyl, oxazolyl and imidazolyl. The heteroaromatic group can be unsubstituted or substituted. The substituents of the heteroaromatic group are selected from halogen, nitro, amino, cyano, hydroxyl, C l ~C6 alkyl, C l ~C6 alkoxy, halogenated C l ~C6 alkyl, halogenated C l ~C6 alkoxy, C3~C6 cycloalkyl, halogenated C3~C6 cycloalkyl.

[0008] As used herein, the term "heterocycloalkyl" refers to a monocyclic or fused ring group having 5 to 9 ring atoms (atoms connecting the radicals to form the ring), one or two of which are heteroatoms selected from N, O or S(O)m (wherein m is an integer from 0 to 2), and the remaining ring atoms are C. These rings may have one or more double bonds, but these rings do not have a completely conjugated π electron system. The heterocycle may be unsubstituted or substituted. Substituted heterocycloalkyls may be pyrrolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino, homopiperazinyl, and the like. Substituents of the heterocycle may be selected from halogen, nitro, amino, cyano, hydroxyl, C l ~C6 alkyl, C l ~C6 alkoxy, halogenated C l ~C6 alkyl, halogenated C l ~C6 alkoxy, C3~C6 cycloalkyl, halogenated C3~C6 cycloalkyl.

[0009] As used herein, the term "cycloalkyl" refers to a saturated monocyclic carbocyclic ring having 3 to 6 carbon atoms, unless a different number of atoms is specified. Cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. "Cycloalkyl" also includes substituted cycloalkyl groups. Cycloalkyl groups may also be optionally substituted on any available carbon with one or more substituents selected from alkoxy, halogen, and haloalkyl, such as perfluoroalkyl.

[0010] The term "alkyl" as used herein refers to an alkyl group having 1 to 6 carbon atoms. It includes but is not limited to methyl, ethyl, propyl, isopropyl, butyl, pentyl, and heptyl. Substituents are selected from halogen, nitro, amino, cyano, hydroxyl, C l ~C6 alkyl, C l ~C6 alkoxy, halogenated C l ~C6 alkyl, halogenated C l ~C6 alkoxy, C3~C6 cycloalkyl, halogenated C3~C6 cycloalkyl.

[0011] As used herein, the term "alkoxy" refers to an -O-alkyl group, wherein alkyl is as defined above. Examples of "alkoxy" as used herein include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, and tert-butoxy. "Alkoxy" also includes substituted alkoxy groups. Alkoxy groups may be substituted one or more times with halogen.

[0012] The term "halogen" as used herein means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine or bromine.

[0013] The term "pharmaceutically acceptable derivatives" refers to salts and solvates of the selected compounds.

[0014] As used herein, the term "solvate" refers to a complex of variable stoichiometry formed by a solute (e.g., a compound of Formula (I) to Formula (VI) of the present invention) and a solvent. For the purposes of the present invention, the solvent must not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid. Preferably, the solvent used is a pharmaceutically acceptable solvent. Suitable pharmaceutically acceptable solvents include, but are not limited to, water, ethanol, and acetic acid. More preferably, the solvent used is water.

[0015] The present invention adopts the following technical solutions:

[0016] The substituted piperidine-heteroaryl derivatives provided by the present invention have the general formula (I):

[0017]

[0018] and its optical isomers or pharmaceutically acceptable salts or solvates,

[0019] in:

[0020] Ring A is selected from unsubstituted or substituted five-membered or six-membered aryl groups, containing 1 to 4 five-membered or six-membered heterocyclic aryl groups selected from O, N, and S;

[0021] R1 is selected from unsubstituted or substituted aryl, unsubstituted or substituted heterocyclic aryl, unsubstituted or substituted cycloalkyl, unsubstituted or substituted saturated or unsaturated heterocyclic alkyl, and any condensed aryl or heterocyclic aryl;

[0022] R2 is selected from amino, cyano, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, olefin-substituted alkyl, C1-C4 carbonyloxy, halogenated C1-C4 alkoxy, wherein n is an integer of 0-4, Rd is selected from H, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, Re is selected from C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, and ring D is selected from unsubstituted or substituted five-membered to eight-membered saturated or unsaturated aliphatic nitrogen-containing heterocyclic ring, unsubstituted or substituted five-membered to eight-membered N-containing heteroaryl, and the carbon atoms in the heterocyclic group and heteroaryl may be further substituted by O or S;

[0023] R3 is selected from H, halogen, hydroxyl, hydroxymethyl, hydroxyethyl, carboxyl, saturated or unsaturated C1-C4 hydrocarbon group, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclic aryl, unsubstituted or substituted saturated or partially saturated heterocycle, unsubstituted or substituted cycloalkyl;

[0024] R4 is selected from H, halogen, nitro, amino, cyano, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy;

[0025] m=an integer from 0 to 3;

[0026] X is selected from O, NH, S, C(═O);

[0027] Y is selected from carbon (CH2), C (=O), sulfonyl;

[0028] The substituted substituent is selected from halogen, nitro, amino, cyano, hydroxyl, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, and halogenated C1-C3 alkoxy.

[0029] Furthermore, the preferred compounds of the present invention have the structures shown in the general formula (II) and (II'):

[0030]

[0031] and its optical isomers or pharmaceutically acceptable salts or solvates,

[0032] in:

[0033] (R1R2R3R4XYm) is as defined by the general formula (I);

[0034] E is selected from (O, S, NH, NH and NCH3);

[0035] T is selected from CH and N.

[0036] Furthermore, the preferred compounds of the present invention have the general formula (III):

[0037]

[0038] and its optical isomers or pharmaceutically acceptable salts or solvates,

[0039] in:

[0040] R1, R2, R3, R4, X, Y, and m are as defined in the general formula (I).

[0041] Furthermore, the preferred compounds of the present invention have the general formula (IV):

[0042]

[0043] and its optical isomers or pharmaceutically acceptable salts or solvates,

[0044] in:

[0045] R1, R2, R3, X, and Y are as defined in the general formula (I);

[0046] E is preferably S.

[0047] As further preferred, X is O or CH; Y is CH2;

[0048] The R1 is selected from substituted phenyl; the substituent is one or more halogen (more preferably one or more F, Cl, Br), halogenated C1-C3 alkyl (methyl, ethyl, propyl or isopropyl);

[0049] R2 is selected from H, 3-(dimethylamino)propyl, allyl, 3-(1H-imidazol-1-yl)propyl, 3-hydroxypropyl, 3-(pyrrolidin-1-yl)propyl, 3-(piperidin-1-yl)propyl, 3-morpholinopropyl, 3-(4-hydroxypiperidin-1-yl)propyl, 2-(methylamino)-2-oxoethyl;

[0050] R3 is selected from H, halogen, C1-C5 alkyl (preferably methyl, ethyl, propyl or isopropyl);

[0051] The R3 is selected from H; E is S.

[0052] More specifically, the preferred compounds of the general formula (IV) of the present invention are:

[0053] 10-Bromo-N-(((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0054] 10-Chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide

[0055] N-((3S,4S)-4-(3,4-Difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide

[0056] 10-Chloro-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide

[0057] 10-Bromo-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide

[0058] N-((3S,4S)-4-(3,5-Difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide

[0059] 10-Bromo-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolino[1,5-d]thiopheno[3,2-f][1,4]oxazepine-2-carboxamide

[0060] 10-Bromo-N-(((3S,4S)-4-(4-chlorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0061] 10-Bromo-N-(((3S,4S)-4-(4-chloro-3-trifluoromethylphenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0062] 10-Bromo-N-(((3S,4S)-4-(3-chloro-4-trifluoromethylphenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0063] 10-Bromo-N-(((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0064] 10-Chloro-N-(((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0065] 1-Bromo-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0066] 1-Bromo-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0067] 1-Bromo-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0068] 1-Chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0069] N-((3S,4S)-4-(3,4-Difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0070] 1-Bromo-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0071] 1-Chloro-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0072] N-((3S,4S)-4-(3-Fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide

[0073] 10-Bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(dimethylamino)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0074] N-((3S,4S,6R)-6-Allyl-4-(3,4-difluorophenyl)piperidin-3-yl)-10-bromo-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0075] N-((3S,4S,6R)-6-(3-(1H-imidazol-1-yl)propyl)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-bromo-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0076] 10-Bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-hydroxypropyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0077] 10-Bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(pyrrolidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0078] 10-Bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(piperidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0079] 10-Bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-morpholinopropyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0080] 10-Bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(4-hydroxypiperidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0081] 10-Bromo-N-(((3S,4S,6S)-4-(3,4-difluorophenyl)-6-(2-(methylamino)-2-oxoethyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0082] 10-Chloro-N-(((3S,4S,6S)-4-(3,4-difluorophenyl)-6-(2-(methylamino)-2-oxoethyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0083] N-((3S,4S)-4-(3,4-Difluorophenyl)piperidin-3-yl)-10-ethyl-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0084] N-((3S,4S)-4-(3,4-Difluorophenyl)piperidin-3-yl)-10-(2-hydroxyethyl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0085] 10-Bromo-N-(((3R,4R)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide

[0086] N-((3S,4S)-4-(3,4-Difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide

[0087] 10-Chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide

[0088] 10-Bromo-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide

[0089] N-((3S,4S)-4-(3,4-Difluorophenyl)piperidin-3-yl)-10-methyl-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide

[0090] and optical isomers of the above compounds or pharmaceutically acceptable salts or solvates thereof.

[0091] More specifically, the preferred compounds of the general formula (IV) of the present invention are:

[0092]

[0093]

[0094]

[0095] In addition, preferred compounds of the present invention have the structure of formula (V):

[0096]

[0097] and its optical isomers or pharmaceutically acceptable salts or solvates,

[0098] in:

[0099] R1, R2, R3, R4, X, and Y are as defined in the general formula (I);

[0100] Furthermore, the preferred compounds of the present invention have the general formula (VI):

[0101]

[0102] and its optical isomers or pharmaceutically acceptable salts or solvates, wherein:

[0103] R1, R2, R3, X, and Y are as defined in the general formula (I);

[0104] More specifically, the preferred compounds of the general formula (VI) of the present invention are:

[0105] 1-Chloro-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide

[0106] N-(((3S,4S)-4-(3,4-Difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide

[0107] N-(((3S,4S)-4-(3-Fluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide

[0108] N-(((3S,4S)-4-(3,5-Difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide

[0109] N-(((3S,4S)-4-(3,4,5-Trifluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide

[0110] and optical isomers of the above compounds or pharmaceutically acceptable salts or solvates thereof.

[0111] The salts of the substituted piperidine-nitrogen heterocyclic compounds of the present invention can be prepared using methods well known to those skilled in the art. The salts can be organic acid salts or inorganic acid salts, such as citrates, fumarates, oxalates, malates, lactates, camphorsulfonates, p-toluenesulfonates, and methanesulfonates. Inorganic acid salts include hydrohalides, sulfates, phosphates, and nitrates. For example, methanesulfonates and trifluoromethanesulfonates can be formed with lower alkylsulfonic acids such as methanesulfonic acid and trifluoromethanesulfonic acid; p-toluenesulfonates and benzenesulfonates can be formed with arylsulfonic acids such as benzenesulfonic acid or p-toluenesulfonic acid; corresponding salts can be formed with organic carboxylic acids such as acetic acid, fumaric acid, tartaric acid, oxalic acid, maleic acid, malic acid, succinic acid, or citric acid; and glutamate or aspartate can be formed with amino acids such as glutamic acid or aspartic acid. Corresponding salts can also be formed with inorganic acids, such as hydrohalic acids (such as hydrofluoric acid, hydrobromic acid, hydroiodic acid, hydrochloric acid), nitric acid, carbonic acid, sulfuric acid or phosphoric acid.

[0112] The second object of the present invention is to provide a pharmaceutical composition comprising at least one active ingredient and one or more pharmaceutically acceptable carriers or excipients, wherein the active ingredient may be any one or more of the substituted piperidine-nitrogen heterocyclic compound of the present invention, an optical isomer of the compound, a pharmaceutically acceptable salt of the compound or its optical isomer, or a solvate of the compound or its optical isomer.

[0113] The carrier includes conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field, and flavoring agents, sweeteners, etc. may also be added if necessary. The drug of the present invention can be prepared in various forms such as tablets, powders, granules, capsules, oral solutions, and injections. The drugs in these dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0114] The present invention also provides the use of the compounds of general formula (I) to general formula (VI), and optical isomers thereof, or pharmaceutically acceptable salts or solvates thereof, in the preparation of anti-tumor drugs. The tumors are neuroblastoma, breast cancer, sarcoma, lung cancer, prostate cancer, colon cancer, rectal cancer, kidney cancer, pancreatic cancer, blood cancer, glioma, head cancer, neck cancer, thyroid cancer, pancreatic cancer, liver cancer, ovarian cancer, vulvar cancer, cervical cancer, endometrial cancer, testicular cancer, bladder cancer, esophageal cancer, gastric cancer, nasopharyngeal cancer, cheek cancer, oral cancer, gastrointestinal stromal cancer, skin cancer, or multiple myeloma.

[0115] The present invention also provides a method for preparing compounds of general formula (I) and pharmaceutically acceptable derivatives thereof. The compounds of general formula (I) are prepared by reacting two main components of the compound, referred to herein as the amine fragment and the acid fragment of the compound. The synthetic routes of some compounds of general formula (I) are as follows:

[0116] Each compound of general formula (I) can be conveniently prepared by separately preparing two constructs of the compound and then combining these components to form the compound of general formula (I). For convenience, the two constructs are referred to herein as amine fragments and acid fragments. When an amine fragment / acid fragment combination appears in the text, it also refers to the corresponding part.

[0117] The acid fragment of the compound of the present invention is a (ternary cycloaryl)carboxylic acid compound represented by the general formula (VII):

[0118]

[0119] The amine fragment of the compound of the present invention is a substituted piperidine compound containing a protecting group represented by the general formula (VIII):

[0120] Compound acid fragment (VII) and amine fragment (VIII) are combined in the synthetic route shown in the following scheme:

[0121]

[0122] Among them, PG is a common amino protecting group, such as Boc (tert-butyloxycarbonyl), Cbz (benzyloxycarbonyl), Ac (acetyl), etc.

[0123] More specifically, general formula (II) and general formula (V) will be used as representatives for explanation.

[0124] The acids of the general formula (II) and the general formula (V) of the present invention are three-membered, five-membered or six-membered aryl carboxylic acid compounds represented by the center (i-1) and the center (i-2) of the general formula:

[0125]

[0126] wherein E and T are as defined in the general formula (II), i.e., E is selected from (O, S, NH, NH and NCH3), and T is selected from CH and N.

[0127] Wherein R3, R4, X, Y, and m are as defined in the general formula (I), that is, R3 is selected from H, halogen, hydroxyl, hydroxymethyl, carboxyl, saturated or unsaturated C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclic aryl, unsubstituted or substituted saturated or partially saturated heterocycle, unsubstituted or substituted cycloalkyl; R4 is selected from H, halogen, nitro, amino, cyano, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy; m is an integer from 0 to 3; X is selected from O, N, S, carbon; Y is selected from carbonyl, sulfonyl, carbon, etc.

[0128] The amine fragment of the compound of the present invention is a Boc-protected substituted piperidine compound represented by the general formula amine fragment (ii)

[0129]

[0130] Wherein the substituents R1 and R2 are as defined in the general formula (I), that is, R1 is selected from unsubstituted or substituted aryl, unsubstituted or substituted heterocyclic aryl, unsubstituted or substituted cycloalkyl, unsubstituted or substituted saturated or unsaturated heterocyclic alkyl, any condensed aryl, heterocyclic aryl,

[0131] R2 is selected from amino, cyano, C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 carboxyl, halogenated C1-C4 alkoxy, wherein n=an integer of 0-4, Rd is selected from H,

[0132] C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, Re is selected from C1-C4 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, and ring D is selected from unsubstituted or substituted five-membered to eight-membered saturated or unsaturated aliphatic nitrogen-containing heterocyclic ring;

[0133] The compound of the general formula acid fragment (i) and amine fragment (ii) are combined in the synthetic route shown in the following scheme

[0134]

[0135] As shown in the above synthetic route, the product obtained by reacting the acid fragment (i) with the amine fragment (ii) in the presence of a condensing agent (EDCI) is subjected to Boc removal under acidic conditions to obtain compounds of the general formula (II) and (V).

[0136] The preparation methods of the compounds of general formula (III), general formula (IV) and general formula (VI) refer to the preparation methods of the compounds of general formula (II) and general formula (V).

[0137] The present invention also provides the use of the compounds described herein, their optical isomers, or pharmaceutically acceptable salts or solvates thereof, in the preparation of AKT inhibitors, particularly in the preparation of agents for treating cell proliferative diseases. Such cell proliferative diseases include cancer. In other words, the present invention provides the use of substituted piperidine-nitrogen heterocyclic compounds or their pharmaceutically acceptable salts, alone or in combination with other drugs, for the treatment of proliferative diseases (such as cancer). Antitumor drugs that can be used in combination with the compounds provided by the present invention or their pharmaceutically acceptable salts include, but are not limited to, at least one of the following categories: mitotic inhibitors (such as vinblastine, vindesine and vinorelbine); tubulin degradation inhibitors (such as taxol); alkylating agents (such as cisplatin, carboplatin and cyclophosphamide); antimetabolites (5-fluorouracil, tirifuracil, methotrexate, cytarabine and hydroxyurea); insertable antibiotics (such as alexin, mitomycin and bleomycin); enzymes (such as aspartase); topoisomerase inhibitors (such as etoposide and camptothecin); biological response modifiers (such as interferon); and proteasome inhibitors (such as bortezomib).

[0138] The inventors of the present invention have demonstrated through multiple experiments that the compounds of the present invention have a significant inhibitory effect on AKT1, exhibiting potent anti-proliferative effects and potent differentiation-inducing effects on mouse neuroblastoma cell lines such as Neuro2a. Therefore, the compounds of the present invention can be used as differentiation-inducing regulators and / or AKT inhibitors in pharmaceuticals for the treatment of solid tumors or blood cancers associated with abnormal cell proliferation and differentiation in humans or animals. BRIEF DESCRIPTION OF THE DRAWINGS

[0139] Figure 1 This is the differentiation-promoting diagram of Neuro2a in the non-medication group;

[0140] Figure 2 This is a diagram showing the differentiation-promoting effect of compound 1 on Neuro2a. DETAILED DESCRIPTION

[0141] The feasibility of the present invention is illustrated below through examples. Those skilled in the art should understand that according to the teachings of the prior art, modifications or replacements of corresponding technical features still fall within the scope of protection claimed by the present invention.

[0142] Example 1. Synthesis of Intermediates 1 to 3

[0143]

[0144] Step 1. Synthesis of compound 1-2

[0145] Dissolve 5-chloro-2-acylthiophene (1.00 g, 5.52 mmol) in 10.0 mL of tetrahydrofuran, then add triethylamine (587 mg, 5.81 mmol) and ethylene glycol mono-tert-butyl ether (686 mg, 5.80 mmol) sequentially. Allow to react at room temperature for 4 h. After completion of the reaction, monitor by TLC. Quench the reaction mixture into water and extract three times with ethyl acetate. The combined organic layers are washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product is purified by silica gel column chromatography to yield 1.21 g of the desired product (Compound 1-2) as a light yellow oil in an 83.7% yield. 1 H NMR (400MHz, CDCl3) δ7.60 (d, J = 4.0Hz, 1H), 6.93 (d, J = 4.0Hz, 1H), 4.40-4.32 (m, 2H), 3.69-3.61 (m, 2H), 1.21 (s, 9H). ESI-MS: m / z = 285.0 [M+Na] + .

[0146] Step 2. Synthesis of Compound 1-3

[0147] Suspend 60% NaH (274 mg, 6.85 mmol) in 10.0 mL of anhydrous DMF (N,N-dimethylformamide). Under nitrogen, add ethylene glycol mono-tert-butyl ether (800 mg, 6.77 mmol) and compound 1-2 (1.20 g, 4.56 mmol) in sequence. Heat to 60°C and react for 2 h. After TLC analysis, pour the reaction solution into 200 mL of water and extract three times with ethyl acetate. Combine the organic layers, wash twice with saturated sodium chloride, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The resulting crude product is quickly purified by silica gel column chromatography to obtain 594 mg of the target product (compound 1-3) as a light yellow oil in a 37.5% yield. 1 HNMR(400MHz, CDCl3)δ7.53(d,J=4.2Hz,1H),6.24(d,J=4.3Hz,1H),4.36-4.29(m,2H),4.22–4.15(m,2 H),3.71(dd,J=5.7,4.3Hz,2H),3.67–3.60(m,2H),1.22(s,9H),1.21(s,9H).ESI-MS:m / z=345.2[M+H] +

[0148] Step 3. Synthesis of Compound 1-4

[0149] Compound 1-3 (590 mg, 1.72 mmol) was dissolved in 10.0 mL of methanol. Under nitrogen, a 50% solution of sodium methoxide in methanol (204 mg, 1.88 mmol) was added. The mixture was heated to reflux and allowed to react for 2 h. After cooling to room temperature, the reaction solution was poured into water and extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was rapidly purified by silica gel column chromatography to yield 430 mg of the target product, compound 1-4, as a colorless oil in a 97% yield; ESI-MS: m / z = 259.1 [M+H] + .

[0150] Step 4. Synthesis of Compound 1-5

[0151] Compound 1-4 (439 mg, 1.70 mmol) was dissolved in 10.0 mL of tetrahydrofuran, and NBS (N-bromosuccinimide, 363 mg, 2.04 mmol) was added. After reacting at room temperature for 1 hour, the reaction was monitored by TLC. After completion, the reaction solution was poured into water to quench the reaction, extracted three times with ethyl acetate, and the organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography to obtain compound 1-5 as a light yellow oily liquid (516 mg, 1.53 mmol) in a yield of 90.0%. ESI-MS: m / z = 359.0 [M+Na]+ .

[0152] Step 5. Synthesis of Compound 1-6

[0153] Compound 1-5 (4.70 g, 14.0 mmol) and 3-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)-1H-pyrazole (3.00 g, 15.5 mmol) were dissolved in a mixture of dioxane (50.0 mL) and water (12.0 mL). Under nitrogen, sodium bicarbonate (3.00 g, 35.7 mmol) and bis(tri-tert-butylphosphine)palladium (1.00 g, 1.96 mmol) were added. The mixture was heated to 86°C and allowed to react for 1 h. TLC monitored the reaction completion. After cooling to room temperature, the reaction solution was poured into water to quench the reaction. The mixture was extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to afford compound 1-6 as a pale yellow solid (3.11 g, 9.60 mmol) in a yield of 68.6%. 1 HNMR (400MHz, CDCl3) δ7.83 (s, 1H), 7.62 (d, J = 2.0Hz, 1H), 6.48 (d, J = 1.9Hz, 1H), 4.4 1–4.33(m,2H),3.88(s,3H),3.84–3.76(m,2H),1.31(s,9H).ESI-MS:m / z=325.1[M+H] + .

[0154] Step 6. Synthesis of Compound 1-7

[0155] Compound 1-6 (500 mg, 1.54 mmol) was dissolved in 5.00 mL of methanol and concentrated hydrochloric acid (1.00 mL) was slowly added dropwise. The mixture was heated to 60°C and allowed to react overnight. TLC confirmed the complete reaction. The reaction solution was poured into water and saturated sodium bicarbonate solution was added dropwise until the pH reached 8-9. The mixture was extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was added with 10 mL of DCM (dichloromethane), stirred at room temperature for 0.5 h, and filtered to obtain compound 1-7 as a white solid (298 mg, 1.11 mmol). The yield was 72.1%. 1 H NMR (400MHz, DMSO) δ10.13(s,2H),8.18(s,1H),7.97(d,J=2.3Hz,1H),6.88(d,J=2.3Hz, 1H), 4.32 (t, J=4.4Hz, 2H), 3.84 (t, J=4.4Hz, 2H), 3.83 (s, 3H). ESI-MS: m / z=269.1[M+H] + .

[0156] Step 7. Synthesis of Compound 1-8

[0157] Compound 1-7 (200 mg, 0.746 mmol) was dissolved in a mixture of chloroform (3.00 mL) and DMA (11.00 mL). Pyridine (60.0 mg, 0.759 mmol) and thionyl chloride (268 mg, 2.25 mmol) were added sequentially under ice-cooling conditions. The reaction was continued under ice-cooling for 30 minutes, then heated to 60°C for 1 hour. TLC monitored the reaction completion. After cooling to room temperature, the reaction mixture was poured into water for quenching and extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to afford compound 1-8 as a pale yellow solid (187 mg, 0.654 mmol) in an 87.6% yield. 1 H NMR (400MHz, CDCl3) δ7.92(s,1H),7.62(d,J=2.0Hz,1H),6.57(d,J=2.0Hz,1H),4 .50(t,J=5.3Hz,2H),3.94(t,J=5.3Hz,2H),3.88(s,3H).ESI-MS:m / z=287.0[M+H] + .

[0158] Step 8. Synthesis of Compound 1-9

[0159] Compound 1-8 (87 mg, 0.304 mmol) was dissolved in 5.00 mL of DMF, and cesium carbonate (195 mg, 0.598 mmol) and potassium iodide (75.5 mg, 0.455 mmol) were added sequentially. The mixture was stirred at room temperature overnight. TLC confirmed the complete reaction. After cooling to room temperature, the reaction mixture was poured into water to quench the reaction. The mixture was extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound 1-9 as a yellow solid (58.52 mg, 0.234 mmol) in a 77% yield. 1 H NMR (400MHz, CDCl3) δ7.72(s,1H),7.48(d,J=2.1Hz,1H),6.43(d,J=2.1Hz,1H ),4.79–4.72(m,2H),4.63–4.56(m,2H),3.87(s,3H).ESI-MS: m / z=251.0[M+H] + .

[0160] Step 9. Synthesis of Compound 1-10a

[0161] Synthesis steps Referring to step 4 of Example 1, a method similar to that of compound 1-5 was used to prepare compound 1-10a with a yield of 89.8%; 1 H NMR (400MHz, CDCl3) δ8.44(s,1H),7.46(s,1H),4.72–4.63(m,2H),4.58(dt,J=5.8,1.8Hz,2H),3.88(s,3H).ESI-MS:m / z=328.7[M+H] + .

[0162] Step 10. Synthesis of Compound 1-10b

[0163] Synthesis steps Referring to step 9 of Example 1, a method similar to that of compound 1-10a was used to prepare compound 1-10b using compound 1-9 and N-chlorosuccinimide (NCS) as raw materials. Yield: 87%; ESI-MS: m / z = 285 [M+H] + .

[0164] Step 11. Synthesis of Intermediate 2

[0165] Compound 1-10a (125 mg, 0.382 mmol) was dissolved in 5.00 mL of a mixture of tetrahydrofuran (THF) and water (v / v, 1:1). Anhydrous lithium hydroxide (91.0 mg, 3.80 mmol) was added, and the mixture was heated to 60°C and refluxed overnight. After TLC analysis, the reaction was complete, and the mixture was concentrated under reduced pressure. 20 mL of water was added to the residue, and the mixture was acidified with 1N hydrochloric acid to a pH of 2-3. A large amount of white solid precipitated. The solid was extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting product (Intermediate 2) was used directly in the next step without further purification. 1 HNMR (400MHz, DMSO) δ13.26(s,1H),8.28(s,1H),7.62(s,1H),4.76–4.69(m,2H),4.69–4.62(m,2H).ESI-MS: m / z=312.9[MH] - .

[0166] Step 12. Synthesis of Intermediate 1

[0167] Synthesis steps: Referring to step 11 of Example 1, a method similar to that of intermediate 2 was used to prepare compound intermediate 1 using compound 1-9 as starting material; ESI-MS: m / z = 235.0 [MH] - .

[0168] Step 13. Synthesis of Intermediate 3

[0169] Synthesis steps: Referring to step 11 of Example 1, intermediate 3 was prepared using compound 1-10b as a starting material using a method similar to that of intermediate 2; ESI-MS: m / z = 269.0 [MH] - .

[0170] Example 2 Synthesis of Intermediates 4 to 6

[0171]

[0172] Step 1. Synthesis of compound 2-2

[0173] Compound 2-1 (1.00 g, 4.30 mmol) and bis(triphenylphosphine)palladium chloride (120 mg, 0.171 mmol) were added to a double-necked flask. Under nitrogen, 8.00 mL of water, 700 μL of tetrahydropyrrole, and (propynyloxy)trimethylsilane (650 mg, 5.07 mmol) were injected sequentially. The mixture was reacted in an oil bath at 100°C for 2 h. TLC monitored the complete reaction. After cooling to room temperature, the reaction solution was poured into water to quench the reaction. The mixture was extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a light yellow oily liquid (380 mg, 1.81 mmol) in a 42.2% yield. 1 H NMR (400MHz, CDCl3) δ7.64(d,J=3.9Hz,1H),7.14(d,J=3.9Hz,1H),4.52(s,2H),4 .34(q,J=7.1Hz,2H),1.79(s,1H),1.37(t,J=7.1Hz,3H).ESI-MS:m / z=209.0[MH] - .

[0174] Step 2. Synthesis of compound 2-3

[0175] Compound 2-2 (2.50 g, 11.9 mmol) was dissolved in 50.0 mL of methanol and palladium on carbon (250 mg, 10%) was added. After three hydrogen substitutions, the reaction was hydrogenated at room temperature under approximately 3.8 atmospheres of hydrogen pressure overnight. The filtrate was filtered and concentrated to afford compound 2-3 as a yellow oil (2.20 g, 10.5 mmol) in an 88.2% yield. It was used directly in the next step without further purification. 1HNMR (400MHz, CDCl3) δ7.63(d,J=3.8Hz,1H),6.81(dt,J=3.8,0.9Hz,1H),4.32(q,J=7.1Hz,2H),3.71(t,J=6 .2Hz,2H),2.95(t,J=7.6Hz,2H),1.95(tt,J=7.6,6.3Hz,2H),1.36(t,J=7.1Hz,3H).ESI-MS:m / z=213.0[MH] - .

[0176] Step 3. Synthesis of Compound 2-4

[0177] Compound 2-3 (100 mg, 0.47 mmol) was dissolved in 1.00 mL of DCM under an ice bath. Aluminum trichloride (186.7 mg, 1.40 mmol) and bromine (74.6 mg, 0.467 mmol) in DCM were added under nitrogen. The reaction was allowed to react at room temperature for 2 h. TLC confirmed the complete reaction. The reaction mixture was poured into water and extracted three times with ethyl acetate. The combined organic layers were washed with saturated sodium thiosulfate and saturated brine, dried over anhydrous sodium sulfate, and the solvent was recovered under reduced pressure. The residue was purified by column chromatography to afford compound 2-4 as a yellow oil (134 mg, 0.46 mmol) in a 97.8% yield. 1 H NMR (400MHz, CDCl3) δ7.60(s,1H),4.33(q,J=7.1Hz,2H),3.71(t,J=6.2Hz,2H),2.92(t,J =7.6Hz, 2H), 1.94 (dq, J = 7.7, 6.3Hz, 2H), 1.35 (t, J = 7.1Hz, 3H). ESI-MS: m / z = 292.8 [M+H] + .

[0178] Step 4. Synthesis of Compound 2-5

[0179] Compound 2-4 (1.00 g, 3.42 mmol) was dissolved in 10.0 mL of DMF and 2.0 mL of water. Under nitrogen, 1H-pyrazole-3-boronic acid pinacolyl (794 mg, 4.08 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (250 mg, 0.342 mmol), and cesium carbonate (2.30 g, 7.06 mmol) were added. The reaction mixture was heated to 100°C for 5 h. TLC confirmed the complete reaction. The mixture was cooled to room temperature and poured into water for quenching. The mixture was extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound 2-5 as a yellow solid (258 mg, 0.914 mmol). The yield was 26.7%. 1 H NMR(400MHz, CDCl3)δ7.83(s,1H),7.66–7.60(m,1H),6.76(s,2H),6.48(s,1H),4.36(q,J=7.1Hz,2 H),3.67(s,2H),3.30(t,J=6.2Hz,2H),2.02(s,2H),1.38(t,J=7.1Hz,3H).ESI-MS:m / z=281.1[M+H] + .

[0180] Step 5. Synthesis of Compound 2-6

[0181] Compound 2-5 (100 mg, 0.357 mmol) and carbon tetrabromide (130 mg, 0.392 mmol) were dissolved in 5 mL of DCM under ice conditions. After nitrogen displacement three times, a solution of triphenylphosphine (121.3 mg, 0.462 mmol) in DCM (2.00 mL) was slowly added dropwise. The reaction was continued in an ice bath overnight. TLC confirmed the reaction was complete. The reaction solution was concentrated, and the residue was purified by column chromatography to afford compound 2-6 as a yellow solid (69.3 mg, 0.203 mmol) in a 56.8% yield. 1 H NMR (400MHz, CDCl3) δ7.93(s,1H),7.65(d,J=2.3Hz,1H),6.52(d,J=2.3Hz,1H),5.72(s,1H),4.35(q,J=7.1Hz,2 H),3.47(t,J=6.5Hz,2H),3.32–3.21(m,2H),2.33–2.20(m,2H),1.38(t,J=7.1Hz,3H).ESI-MS:m / z=343.0[M+H] + .

[0182] Step 6. Synthesis of Compound 2-7

[0183] Synthesis steps: Referring to step 8 of Example 1, a method similar to that of compound 1-9 was used, and compound 2-6 was used as the starting material to prepare compound 2-7 with a yield of 82.9%; 1 H NMR (400MHz, CDCl3) δ7.90(s,1H),7.49(d,J=2.0Hz,1H),6.54(d,J=2.0Hz,1H),4.62–4.56(m,2H),4.40(q,J =7.1Hz,2H),3.30–3.24(m,2H),2.36(qt,J=7.0,4.0Hz,2H),1.43(t,J=7.2Hz,3H).ESI-MS:m / z=263.1[M+H] + .

[0184] Step 7. Synthesis of compound 2-8a

[0185] Synthesis steps Referring to step 4 of Example 1, a method similar to that of compound 1-5 was used to prepare compound 2-8a using compound 2-7 and N-bromosuccinimide (NBS) as raw materials with a yield of 95%; 1 H NMR (400MHz, DMSO) δ8.17(s,1H),7.62(s,1H),4.32–4.30(m,2H),3.82–3.75(m,2H),3. 14(t,J=7.0Hz,2H),2.35–2.18(m,2H),1.30(t,J=7.1Hz,3H).ESI-MS:m / z=340.9[M+H] + .

[0186] Step 8. Synthesis of compound 2-8b

[0187] Synthesis steps Referring to step 4 of Example 1, a method similar to that of compound 1-5 was used to prepare compound 2-8b using compound 2-7 and N-chlorosuccinimide (NCS) as raw materials. Yield: 80%; ESI-MS: m / z = 297 [M+H] + .

[0188] Step 9. Synthesis of Intermediate 4

[0189] Synthesis steps: Referring to step 11 of Example 1, intermediate 4 was prepared using a method similar to that of intermediate 2, using compound 2-7 as the starting material; ESI-MS: m / z = 233.0 [MH] - .

[0190] Step 12. Synthesis of Intermediate 5

[0191] Synthesis steps: Referring to step 11 of Example 1, intermediate 5 was prepared using a method similar to that of intermediate 2, using compound 2-8a as the starting material; ESI-MS: m / z = 311.0 [MH] - .

[0192] Step 13. Synthesis of Intermediate 6

[0193] Synthesis steps: Referring to step 11 of Example 1, intermediate 6 was prepared using a method similar to that of intermediate 2, using compound 2-8b as the starting material; ESI-MS: m / z = 267.0 [MH] - .

[0194] Example 3 Synthesis of Intermediates 7-8

[0195]

[0196] Step 1. Synthesis of compound 3-2

[0197] Methyl 4-bromo-3-hydroxybenzoate (500 mg, 2.16 mmol) was dissolved in MeCN (10.0 mL), and cesium carbonate (1.70 g, 5.22 mmol) and 1,2-dibromoethane (3.30 g, 17.6 mmol) were added. The reaction was heated under reflux for 5 h. TLC confirmed the complete reaction. The mixture was cooled to room temperature and poured into water to quench the reaction. The mixture was extracted three times with ethyl acetate. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound 3-2 as a colorless oily liquid (717.1 mg, 2.13 mmol). The yield was 98.6%. 1 H NMR (400MHz, CDCl3) δ7.69 (d, J=8.1Hz, 1H), 7.64–7.57 (m, 2H), 4.46 (td, J=6.4, 3. 3Hz, 2H), 4.01–3.95 (m, 3H), 3.76 (td, J=6.4, 3.3Hz, 2H). ESI-MS: m / z=336.9[M+H] + .

[0198] Step 2. Synthesis of compound 3-3

[0199] Synthesis steps: Referring to step 5 of Example 1, a similar synthesis method as compound 1-6 was used to prepare compound 3-3 using compound 3-2 as the starting material. Yield: 84.2%; ESI-MS: m / z = 325.0 [M+H] + .

[0200] Step 3. Synthesis of compound 3-4

[0201] Synthesis steps: Referring to step 8 of Example 1, a method similar to that of compound 1-9 was used to prepare compound 3-4 using compound 3-3 as the starting material. The yield was 78.4%; ESI-MS: m / z = 245.1 [M+H] + .

[0202] Step 4. Synthesis of compound 3-5

[0203] Synthesis steps Referring to step 4 of Example 1, a method similar to that of compound 1-5 was used to prepare compound 3-5 using compound 3-4 and N-chlorosuccinimide (NCS) as raw materials with a yield of 89.0%; 1 H NMR (400MHz, DMSO) δ7.91 (d, J=8.2Hz, 1H), 7.83 (dd, J=8.2, 1.7Hz, 1H), 7.78 (s, 1H), 7.71 (d, J= 1.6Hz,1H),4.60(t,J=5.1Hz,2H),4.51(t,J=5.1Hz,2H),3.89(s,3H).ESI-MS:m / z=279.1[M+H] + .

[0204] Step 5. Synthesis of Intermediate 8

[0205] Synthesis steps: Referring to step 11 of Example 1, intermediate 8 was prepared using a method similar to that of intermediate 2, using compound 3-5 as the starting material; ESI-MS: m / z = 263.0 [MH] - .

[0206] Step 6. Synthesis of Intermediate 7

[0207] Synthesis steps: Referring to step 11 of Example 1, a similar synthesis method as that of intermediate 2 was used to prepare compound intermediate 7 using compound 3-4 as starting material; ESI-MS: m / z = 243.0 [MH] - .

[0208] Example 4 Synthesis of Intermediates 9 to 11

[0209]

[0210] Step 1 Synthesis of compound 4-2

[0211] Compound 4-1 (2.1 g, 11 mmol) was dissolved in 30 mL of glacial acetic acid and iron powder (3.1 g, 55 mmol) was added. The reaction solution was heated to 50 degrees Celsius for 1 h. TLC detected that the raw material reaction was complete, and LC-MS monitored that the hydroxylamine intermediate was completely converted. 50 ml of saturated sodium bicarbonate was added to quench the reaction and the mixture was extracted with ethyl acetate (500 mL × 3). The organic layer was synthesized and washed twice with saturated NaCl solution and dried over anhydrous sodium sulfate. The filtrate was concentrated to give a yellow oily liquid (2.20 g, 10.5 mmol) compound 4-2 in a yield of 95%. No further purification was required and it was used directly in the next step. ESI-MS: m / z=158.0[M+H] + .

[0212] Step 2 Synthesis of compound 4-3

[0213] Compound 4-2 (1.48 g, 9.43 mmol) and 4-dimethylaminopyridine (115 mg, 0.94 mmol) were dissolved in 20 mL of 1,4-dioxane, and triethylamine (1.14 g, 11.3 mmol) and di-tert-butyl dicarbonate (2.47 g, 11.3 mmol) were added. The reaction solution was heated to 80 degrees Celsius for 6 hours. TLC confirmed the complete reaction of the starting material. 100 mL of water was added and the mixture was extracted with ethyl acetate (300 mL x 3). The organic layer was washed twice with saturated NaCl solution, dried over anhydrous sodium sulfate, and the solvent was recovered under reduced pressure. The residue was purified by column chromatography to obtain compound 4-3 as a pale yellow solid (1.54 g, 6 mmol). The yield was 63.6%. 1 H NMR(400MHz,Chloroform-d)δ7.61(d,J=4.1Hz,1H),6.50(d,J=4.1Hz,1H),3.88(s,3H),1.57(s,9H).ESI-MS:m / z=258.0[M+H] +

[0214] Step 3 Synthesis of compound 4-4

[0215] Synthesis steps: Referring to step 4 of Example 1, a method similar to that of compound 1-5 was used to prepare compound 4-4 using compound 4-3 and N-bromosuccinimide (NBS) as starting materials. The yield was 80%. ESI-MS: m / z = 336.0 [M+H] + .

[0216] Step 4 Synthesis of Compound 4-5

[0217] Synthesis steps: Referring to step 5 of Example 1, a synthesis method similar to that of compound 1-6 was used to prepare compound 4-5 using compound 4-4 as starting material with a yield of 73.9%. 1H NMR (400MHz, Chloroform-d) δ10.66(s,1H),7.93(s,1H),7.67(d,J=2.5Hz,1H),6.57(d,J=2.5Hz,1H),3.91(s,3H),1.60(s,9H).ESI-MS: m / z=324.1[M+H] + .

[0218] Step 5 Synthesis of compounds 4-6 and 4-7

[0219] Compound 4-5 (91 mg, 0.28 mmol) was dissolved in 4N hydrochloric acid ethanol solution (5 mL) and stirred at room temperature for 3 h. TLC confirmed complete conversion of the starting material. The solvent was removed under reduced pressure to obtain crude compound 4-6, which was directly dissolved in 5.0 mL of anhydrous tetrahydrofuran. Under ice bath conditions, triethylamine (85 mg, 0.84 mmol) was added, and chloroacetyl chloride (97 mg, 0.84 mmol) was slowly added dropwise. The reaction was allowed to proceed at room temperature for 30 minutes. TLC monitored the reaction completion. The solvent was removed under reduced pressure, 30 mL of water was added, and the mixture was extracted with ethyl acetate (40 mL × 3). The organic layers were combined, washed twice with saturated NaCl solution, dried over anhydrous sodium sulfate, and the solvent was recovered under reduced pressure. The residue was purified by column chromatography to obtain compound 4-7 as a white solid (44 mg, 0.15 mmol) in a yield of 52%. 1 H NMR (400MHz, DMSO-d6) δ13.33(s,1H),12.58(s,1H),8.17(s,1H),7.93(d,J= 1.9Hz,1H),6.93(s,1H),4.68(s,2H),3.85(s,3H).ESI-MS:m / z=300.0[M+H] + .

[0220] Step 6 Synthesis of Compound 4-8

[0221] Synthesis steps: Referring to step 8 of Example 1, a method similar to that of compound 1-9 was used to prepare compound 4-8 using compound 4-7 as starting material, with a yield of 48.4%. 1 H NMR(400MHz,Chloroform-d)δ7.80(s,1H),7.60(d,J=1.9Hz,1H),6.49(d,J=1.9Hz,1H),4.99(s,2H),3.95(s,3H).ESI-MS:m / z=264.0[M+H] + .

[0222] Step 7. Synthesis of compound 4-9a

[0223] Synthesis steps Referring to step 4 of Example 1, a method similar to that of compound 1-5 was used to prepare compound 4-9a using compound 4-8 and N-bromosuccinimide (NBS) as raw materials with a yield of 97.7%. 1 H NMR (400MHz, DMSO-d6) δ11.65(s,1H),8.03(d,J=5.1Hz,1H),7.80(s,1H),4.99(s,2H),3.89(s,3H).ESI-MS:m / z=342.0[M+H] + .

[0224] Step 8. Synthesis of compound 4-9b

[0225] Synthesis steps Referring to step 4 of Example 1, a method similar to that of compound 1-5 was used to prepare compound 4-9b using compound 4-8 and N-chlorosuccinimide (NCS) as raw materials. The yield was 97%. 1 H NMR (400MHz, DMSO-d6) δ11.67(s,1H),7.96(d,J=5.7Hz,1H),7.80(s,1H),4.98(s,2H),3.88(s,3H).ESI-MS:m / z=298.0[M+H] + .

[0226] Step 9. Synthesis of Intermediate 9

[0227] Synthesis steps Referring to step 11 of Example 1, intermediate 9 was prepared using a synthesis method similar to that of intermediate 2 and compound 4-8 as starting material; ESI-MS: m / z = 248.0 [MH]-.

[0228] Step 12. Synthesis of Intermediate 10

[0229] Synthesis steps: Referring to step 11 of Example 1, intermediate 10 was prepared using a method similar to that of intermediate 2, using compound 4-9a as the starting material; ESI-MS: m / z = 326.0 [MH] - .

[0230] Step 13. Synthesis of Intermediate 11

[0231] Synthesis steps: Referring to step 11 of Example 1, intermediate 11 was prepared using a method similar to that of intermediate 2, using compound 4-9b as the starting material; ESI-MS: m / z = 282.0 [MH] - .

[0232] Example 5 Synthesis of Intermediates 12 to 19

[0233]

[0234] Step 1: Synthesis of Compound 5-3a (Synthesis of Compounds 5-3b to 5-3g, see Table 1)

[0235] Tert-butyl 2-nitroethylcarbamate (compound 5-1, 380 mg, 2 mmol), ((S)-(-)-α,α-diphenyl-2-pyrrolmethyl)trimethylsilyl ether (33 mg, 0.1 mmol) ((S)-2-(diphenyl((trimethylsilyl)oxy)methyl)pyrrolidine), benzoic acid (25 mg, 0.2 mmol) were dissolved in anhydrous dichloromethane (2 ml) and 3,4-difluorocinnamaldehyde (compound 5-2a, 168 mg, 1 mmole) was slowly added under ice bath conditions. ol), stirred at room temperature for about 24 hours, the reaction system was diluted with dichloromethane to more than five times, 200 μl of trifluoroacetic acid was slowly added dropwise to the reaction solution, and the reaction was allowed to react at room temperature for 5 hours. Subsequently, about 10 ml of 1N sodium bicarbonate solution was added to the reaction solution, and stirred at room temperature for 10 minutes. The reaction solution was then extracted three times with ethyl acetate, and the combined organic phases were washed once with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 228 mg of a light yellow solid (compound 5-3a), with a yield of 67%; ESI-MS: m / z=341[M+H] + .

[0236] Table 1 Synthesis method of compounds 5-3b to 5-3g

[0237]

[0238]

[0239] Step 2 Synthesis of Intermediate 12 (Synthesis of Intermediates 13 to 18 is shown in Table 2)

[0240] Compound 5-3a (170 mg, 0.5 mmol) was dissolved in ethyl acetate (10 ml), and 30 mg of 10% Pd / C was added. The mixture was hydrogenated at room temperature overnight. After the reaction was completed, the mixture was filtered and the filtrate was dried to give 101 mg of an oily liquid (Intermediate 12) in a yield of 65%; ESI-MS: m / z = 313 [M+H] + .

[0241] Table 2 Synthesis methods of intermediates 13 to 18

[0242]

[0243] Step 3 Synthesis of Compound 5-4

[0244] Synthesis steps Referring to step 1 of Example 5, a method similar to that of compound 5-3a was used to prepare compound 5-4 using compounds 5-1, 5-2a, and ((R)-(-)-α,α-diphenyl-2-pyrrolylmethyl)trimethylsilyl ether as raw materials; ESI-MS: m / z=341 [M+H] + .

[0245] Step 4 Synthesis of Intermediate 19

[0246] Synthesis steps: Refer to step 2 of Example 5, and use a method similar to that of intermediate 12, using compound 5-4 as the starting material to prepare intermediate 19. ESI-MS: m / z=313 [M+H] + .

[0247] Example 6 Synthesis of Intermediate 20

[0248]

[0249] Dissolve tert-butyl 2-nitroethylcarbamate (compound 6-1, 2.85 g, 15 mmol), ((S)-(-)-α,α-diphenyl-2-pyrrolyl)trimethylsilyl ether (0.36 g, 1.1 mmol), and benzoic acid (0.25 g, 2 mmol) in anhydrous dichloromethane (15 ml). Slowly add 3,4-difluorocinnamaldehyde (compound 6-2, 1.68 g, 10 mmol) under ice bath conditions. Stir at room temperature for about 18 h to generate compound 6-3 and proceed directly to the next step. Dilute the reaction solution with dichloromethane. The mixture was diluted to 100 ml and cooled to -78°C. Allyltrimethylsilane (5 ml, 30 mmol) was added to the reaction solution, followed by slow dropwise addition of boron trifluoride etherate. The reaction was continued for 10 h. 100 ml of 1N sodium bicarbonate solution was added to the reaction solution, and the mixture was stirred at room temperature for 10 min. The mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain 1.9 g of a white solid (Intermediate 20) in a 50% yield; ESI-MS: m / z = 383 [M+H] + .

[0250] Example 7 Synthesis of Intermediate 21

[0251]

[0252] Step 1 Synthesis of Compound 7-1

[0253] Intermediate 19 (149 mg, 0.390 mmol) was dissolved in ethanol (10.0 mL) and water (2.00 mL). Iron powder (110 mg, 1.95 mmol) and ammonium chloride (42.0 mg, 0.785 mmol) were added, and the mixture was heated to reflux with mechanical stirring for 3 h. TLC monitored the reaction for complete reaction. After the ethanol was removed from the reaction solution, the residual liquid was poured into 100 mL of saturated sodium bicarbonate solution and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to yield 102 mg of a light yellow oily liquid (Compound 7-1). ESI-MS: m / z = 296 [M-Tert-butyl] + .

[0254] Step 2 Synthesis of compound 7-2

[0255] Intermediate 2 (110 mg, 0.35 mmol), 1-hydroxybenzotriazole (HOBt) (97 mg, 0.345 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EDCI (98.8 mg, 0.517 mmol) were dissolved in anhydrous dichloromethane (4 ml) and stirred on ice for 10 minutes. Diisopropylethylamine (0.115 ml, 1.21 mmol) was added and continued to stir on ice for 15 minutes. Compound 7-1 (123 mg, 0.35 mmol) was added and stirred at room temperature overnight. After completion of the reaction monitored by TLC, the mixture was poured into 15 ml of water and extracted with dichloromethane three times. The organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and dried by spin drying. The mixture was purified by column chromatography to give 150 mg of a light yellow solid (compound 7-2) in a yield of 66%. ESI-MS: m / z=649 [M+H] + .

[0256] Step 3 Synthesis of compound 7-3

[0257] Compound 7-2 (648 mg, 1 mmol) was dissolved in 5 ml of tetrahydrofuran. Borane was slowly added until the bubbles disappeared. TLC monitored the reaction completion. 3 ml of 4N NaOH solution was slowly added. The residue was poured into 50 ml of water and extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 500 mg of a colorless oily liquid (compound 7-3), in a 75% yield. ESI-MS: m / z = 667 [M+H] + .

[0258] Step 4 Synthesis of Intermediate 21

[0259] Compound 7-3 (666 mg, 1 mmol) was dissolved in 5 mL of dichloromethane solution. DIPEA (mg, 3 mmol) was added under ice-bath conditions, and then methanesulfonyl chloride (212 mg, 1.1 mmol) was slowly added dropwise. The mixture was stirred at room temperature for 20 min. After completion of the reaction monitored by TLC, the mixture was poured into 15 mL of water and extracted with dichloromethane three times. The organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried. The mixture was purified by column chromatography to give 600 mg of a light yellow solid (Intermediate 21) in a yield of 80.6%. ESI-MS: m / z = 745 [M+H] + .

[0260] Example 8 Synthesis of Intermediate 22

[0261]

[0262] Step 1 Synthesis of Compound 8-1

[0263] Intermediate 20 (1.9 g, 5 mmol) was dissolved in 40 ml of a mixed solvent of DCM / CH3CN / H2O (v / v / v=1 / 1 / 2). Under ice-bath conditions, NaIO4 (5.35 g, 25 mmol) and RuCl3 monohydrate (170 mg, 1 mmol) were slowly added in sequence. The mixture was stirred at room temperature overnight, and the black insoluble matter was filtered off. The filtrate was adjusted to pH=5 with dilute hydrochloric acid solution and extracted three times with dichloromethane. The organic layers were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to obtain 1.8 g of a colorless oil (Compound 8-1) with a yield of 90%. 1 HNMR(500MHz,CDC13)δ7.11(dd,J=18.2,8.4Hz,lH),7.05-7.00(m,lH),6.92(dd,J=5.4,3.lHz,lH), 4.89(m,lH),4.62(m,2H),3.42(m,lH),3.29(m,lH),2.76(d,J=6.7Hz,2H),1.97(s,2H),1.46(s,9H).

[0264] Step 2 Synthesis of compound 8-2

[0265] Intermediate 2-3 (200 mg, 0.5 mmol) was dissolved in 5 ml of DMF. HBTU (379 mg, 1 mmol) and triethylamine (0.25 ml) were added sequentially in an ice bath. After 15 minutes of reaction at room temperature, 1 ml of 30% methylamine in ethanol was added dropwise and the reaction continued for 3 hours. Upon completion of the reaction, the mixture was poured into 10 ml of water and extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to obtain 200 mg of a white solid (Intermediate 5-2) in a 97% yield. This solid was used directly in the next step without purification.

[0266] Step 3 Synthesis of Intermediate 22

[0267] Synthesis steps: Referring to step 1 of Example 7, similar to the synthesis method of compound 7-1, intermediate 22 was prepared using compound 8-2 as the starting material with a yield of 91%; ESI-MS: m / z=384 [M+H] + .

[0268] Example 10 Synthesis of Intermediates 24-25

[0269]

[0270] Step 1 Synthesis of Intermediate 24

[0271] Synthesis steps: Referring to step 2 of Example 7, intermediate 12 and intermediate 2 were used as raw materials to prepare intermediate 24 with a yield of 63%; ESI-MS: m / z = 609 [M+H] + .

[0272] Step 2 Synthesis of Intermediate 25

[0273] Tris(dibenzylideneacetone)dipalladium (Pd2(dba)3) (91.6 mg, 0.1 mmol) and 10% tri-tert-butylphosphine pentanediol solution (0.8 mL, 0.4 mmol) were dissolved in 3 mL of anhydrous DMF and stirred at room temperature for 3 h under N2 protection. 20 mL of a mixed solution of dioxane and water (v / v, 5:1) was added dropwise to the reaction solution to dilute it. The mixture was cooled to 0°C and intermediate 24 (608 mg, 1 mmol), ethylene boronic anhydride pyridine complex (265 mg, 1.1 mmol) and potassium carbonate (414 mg, 3 mmol) were added. The mixture was heated to 70°C and stirred overnight. After the reaction of the starting material was complete, the reaction solution was poured into water (50 mL) and extracted with EA (30 mL × 3). The organic layers were combined, washed with saturated NaCl solution (30 mL × 2), dried over anhydrous sodium sulfate, and the solvent was recovered under reduced pressure. The residue was purified by column chromatography to obtain intermediate 25 as a white solid (278 mg, 0.5 mmol). ESI-MS: m / z = 557 [M+H] + .

[0274] Example 11 Synthesis of Intermediate 26

[0275]

[0276] Step 1 Synthesis of compound 10-2

[0277] Compound 10-1 (328.44 mg, 4 mmol) was dissolved in 5 ml of tetrahydrofuran, and 3,4-dihydro-2H-pyran (328.44 mg, 4 mmol) and 40 μl of trifluoroacetic acid were added. The mixture was stirred overnight in an oil bath at 80°C. After completion of the reaction, the mixture was poured into 15 ml of water and extracted three times with EA. The organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and then purified by column chromatography to afford 400 mg of compound 10-2 as a pale yellow solid in a yield of 60.25%. ESI-MS: m / z = 167 [M+H] + .

[0278] Step 2 Synthesis of compound 10-3

[0279] Compound 10-2 (1 g, 6.024 mmol) was dissolved in 10 ml of anhydrous tetrahydrofuran. The reaction solution was cooled to -40 °C and n-butyllithium (424 mg, 7.23 mmol, 3 ml) (2.5 mol / L in haxane) was added dropwise. The mixture was stirred at this temperature for 1 h. 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.23 g, 6.626 mmol) was then added dropwise. The mixture was stirred at room temperature. After completion of the reaction, the reaction was quenched by TLC. The pH was adjusted to 5-6 and the mixture was extracted with EA. The organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and spin-dried to give 1.557 g of compound 10-3 as a yellow oily liquid in a yield of 88.5%. The compound was used directly in the next step. ESI-MS: m / z=293 [M+H] + .

[0280] Step 3 Synthesis of compound intermediate 10-4

[0281] Compound 10-3 (200 mg, 0.685 mmol) was dissolved in 2 ml of methanol and gradually added to 2 ml of 4N hydrochloric acid methanol solution under ice bath. After stirring overnight, TLC plate was observed. After the reaction was completed, the reaction solution was spin-dried to obtain 170 mg of intermediate 10-4 as a light yellow oil. ESI-MS: m / z = 127 [M+H] + .

[0282] Step 4 Synthesis of compound 10-5

[0283] Synthesis Steps Referring to step 5 of Example 1, a method similar to that of compound 1-6 was used to prepare compound 10-5 using compounds 1-5 and 10-4 as raw materials.

[0284] Step 5 Synthesis of compound 10-6

[0285] Synthesis Steps Referring to step 6 of Example 1, a synthesis method similar to that of compound 1-7 was used to prepare compound 10-6 using compound 10-5 as a raw material.

[0286] Step 6 Synthesis of compound 10-7

[0287] Synthesis Steps Referring to step 7 of Example 1, a method similar to that of compound 1-8 was used to prepare compound 10-7 using compound 10-6 as a raw material.

[0288] Step 7 Synthesis of compound 10-8

[0289] Synthesis Steps Referring to step 8 of Example 1, a synthesis method similar to that of compound 1-9 was used to prepare compound 10-8 using compound 10-7 as a raw material.

[0290] Step 8 Synthesis of Intermediate 26

[0291] Synthesis steps Referring to step 10 of Example 1, intermediate 26 was prepared using a synthesis method similar to that of intermediate 2 and compound 10-8 as a starting material.

[0292] Example 12 Synthesis of 10-bromo-N-(((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 1)

[0293]

[0294] To the obtained intermediate 24 was added 2 ml of 4N hydrochloric acid-ethanol solution, and the mixture was stirred at room temperature for two hours, followed by TLC analysis. After completion of the reaction, the mixture was spin-dried, saturated sodium bicarbonate was added, and the mixture was extracted twice with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain a white solid product (Compound 1) in an 87% yield. 1H NMR (400MHz, DMSO-d6) δ9.53 (s, 1H), 9.41 (s, 1H), 8.76 (d, J = 8.8Hz, 1H), 8.08 (s, 1H), 7.62 (s, 1H) ,7.44–7.27(m,2H),7.18–7.09(m,1H),4.77–4.65(m,2H),4.59(dtd,J=13.5,11.0,9.8,6.2Hz,2H ),4.43(dtd,J=15.6,11.5,10.0,4.2Hz,1H),3.39(d,J=11.8Hz,2H),3.17(td,J=11.6,4.0Hz,1H) ,2.95(q,J=10.0,9.1Hz,2H),2.16–2.05(m,1H),2.01(d,J=12.5Hz,1H).ESI-MS:m / z=509.0[M+H] + .

[0295] Example 13. Synthesis of 10-chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolino[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide (Compound 2)

[0296]

[0297] Intermediate 3 (92.4 mg, 0.35 mmol), 1-hydroxybenzotriazole (HOBt) (97 mg, 0.345 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EDCI (98.8 mg, 0.517 mmol) were dissolved in anhydrous dichloromethane (4 ml), stirred under ice bath for 10 minutes, diisopropylethylamine (0.115 ml, 1.21 mmol) was added, and continued to stir under ice bath for 15 minutes, and intermediate 12 (110 mg, 0.35 mmol) was added, and stirred at room temperature overnight. After the reaction was completed, it was poured into 15 ml of water, and the reaction solution was extracted with dichloromethane 3 times. The organic phases were combined, washed with saturated sodium chloride 2 times, dried over anhydrous sodium sulfate, spin-dried, and purified by column chromatography to obtain a light yellow solid; 2 ml of the obtained light yellow solid was added 4N hydrochloric acid ethanol solution was stirred at room temperature for two hours and then TLC plate was observed. After the reaction was completed, it was dried by spin drying, saturated sodium bicarbonate was added, and extracted twice with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain a white solid product (Compound 2) with a yield of 88.2%; the yield was 34%. 1H NMR (500MHz, DMSO-d6) δ9.46(d,J=10.6Hz,1H),9.34(d,J=12.0Hz,1H),8.75(d,J=8.9Hz,1H),8.01( s,1H),7.59(s,1H),7.34(dt,J=10.7,8.5Hz,1H),7.31–7.24(m,1H),7.09(dt,J=9.6,2.9Hz,1H),4.6 5(t,J=4.2Hz,2H),4.61–4.51(m,2H),4.48–4.30(m,1H),3.41–3.31(m,2H),3.14(td,J=11.6,4.1Hz, 1H),3.00–2.85(m,2H),2.05(dd,J=13.0,3.6Hz,1H),1.99(q,J=4.9Hz,1H).ESI-MS:m / z=465.0[M+H] + .

[0298] Example 14. Synthesis of N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide (Compound 3)

[0299]

[0300] Synthesis steps: Referring to Example 13, intermediate 12 and intermediate 1 were used as raw materials to prepare compound 3 with a yield of 34%. 1 H NMR(400MHz,DMSO-d6)δ9.93(s,1H),9.73(s,1H),9.05(d,J=8.9Hz,1H),8.50(d,J=4.8Hz,1H), 8.21(s,1H),7.52–7.43(m,1H),7.33(dd,J=8.3,4.5Hz,2H),6.50(s,1H),4.62(d,J=4.4Hz,2H) ,4.55(t,J=3.9Hz,2H),3.35(d,J=6.5Hz,2H),3.20(dd,J=11.2,4.2Hz,1H),3.00(d,J=12.5Hz, 1H),2.93(d,J=12.0Hz,2H),2.15(q,J=12.9Hz,1H),1.98–1.87(m,1H).ESI-MS:m / z=431.0[M+H] + .

[0301] Example 15. Synthesis of 10-chloro-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide (Compound 4)

[0302]

[0303] Synthesis steps: Referring to Example 13, intermediate 14 and intermediate 3 were used as raw materials to prepare compound 4 with a yield of 34%. 1 H NMR(500MHz,DMSO-d6)δ9.46(d,J=10.6Hz,1H),9.34(d,J=11.0Hz,1H),8.71(d,J=9.0Hz,1H), 8.00(s,1H),7.59(s,1H),7.31(td,J=8.0,6.3Hz,1H),7.16–7.04(m,2H),7.01(td,J=8.6,2.6 Hz,1H),4.71–4.61(m,2H),4.60–4.51(m,2H),4.48–4.38(m,1H),3.36(dt,J=12.2,3.1Hz,2H) ,3.14(td,J=11.7,4.2Hz,1H),3.01–2.86(m,2H),2.12–1.93(m,2H).ESI-MS:m / z=447.0[M+H] + .

[0304] Example 16. Synthesis of 10-bromo-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide (Compound 5)

[0305]

[0306] Synthesis steps: Referring to Example 13, intermediate 14 and intermediate 2 were used as raw materials to prepare compound 5 with a yield of 45%. 1H NMR(500MHz,DMSO-d6)δ9.52(d,J=10.6Hz,1H),9.38(d,J=11.2Hz,1H),8.69(d,J=8.7Hz,1H),8.03(s,1H), 7.59(s,1H),7.36–7.28(m,1H),7.18–7.05(m,2H),7.01(td,J=8.6,2.5Hz,1H),4.66(t,J=5.3Hz,2H),4.61 –4.49(m,2H),4.44(ddt,J=16.3,11.4,5.7Hz,1H),3.35(dd,J=11.1,3.5Hz,2H),3.14(td,J=11.8,3.8Hz,1 H),2.92(dt,J=22.4,11.1Hz,2H),2.06(td,J=12.9,3.5Hz,1H),2.01–1.94(m,1H).ESI-MS:m / z=491.0[M+H] + .

[0307] Example 17. Synthesis of N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide (Compound 6)

[0308]

[0309] Synthesis steps: Referring to Example 13, intermediate 12 and intermediate 1 were used as raw materials to prepare compound 6 with a yield of 48%. 1 H NMR (500MHz, Methanol-d4) δ7.55 (s, 1H), 7.42 (d, J = 2.0Hz, 1H), 6.96–6.87 (m, 2H), 6.73 (tt, J = 9. 1,2.3Hz,1H),6.41(d,J=2.1Hz,1H),4.66–4.60(m,2H),4.60–4.53(m,2H),4.24(td,J=11.1,4.5Hz ,1H),3.31–3.23(m,1H),3.19–3.11(m,1H),2.91(td,J=11.8,3.8Hz,1H),2.73(td,J=12.8,2.8Hz, 1H),2.65(dd,J=12.4,10.9Hz,1H),1.97–1.90(m,1H),1.86–1.71(m,1H).ESI-MS:m / z=430.9[M+H] + .

[0310] Example 18. Synthesis of 10-bromo-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolino[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide (Compound 7)

[0311]

[0312] Synthesis steps: Referring to Example 13, intermediate 12 and intermediate 2 were used as raw materials to prepare compound 7 with a yield of 43%. 1 H NMR (400MHz, DMSO-d6) δ9.50(d,J=10.5Hz,1H),9.37(d,J=11.6Hz,1H),8.76(d,J=8.8Hz,1H),8. 06(s,1H),7.59(s,1H),7.07(tt,J=9.4,2.4Hz,1H),6.98(h,J=4.1Hz,2H),4.74–4.64(m,2H),4.6 3–4.51(m,2H),4.42(qd,J=11.4,4.3Hz,1H),3.37(td,J=7.9,7.5,4.3Hz,2H),3.19(td,J=11.6,4 .2Hz,1H),2.93(p,J=11.0,10.5Hz,2H),2.16–2.03(m,1H),2.02(s,1H).ESI-MS:m / z=509.0[M+H] + .

[0313] Example 19. Synthesis of 10-bromo-N-(((3S,4S)-4-(4-chlorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 8)

[0314]

[0315] Synthesis steps: Referring to Example 13, intermediate 15 and intermediate 2 were used as raw materials to prepare compound 8 with a yield of 50%. 1H NMR (400MHz, Methanol-d4) δ8.08(d,J=5.2Hz,1H),7.64(d,J=7.7Hz,2H),7.56(dd,J=8.1,5.2Hz,2H),7.49(d,J=5.2Hz,1H),4.71 –4.64(m,2H),4.63–4.53(m,3H),3.69–3.50(m,3H),3.28–3.14(m,2H),2.22(s,1H),2.16(d,J=11.5Hz,1H).ESI-MS:m / z=507[M+H] + .

[0316] Example 20. Synthesis of 10-bromo-N-(((3S,4S)-4-(4-chloro-3-trifluoromethylphenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 9)

[0317]

[0318] Synthesis steps: Referring to Example 13, intermediate 16 and intermediate 2 were used as raw materials to prepare compound 9 with a yield of 51%. 1 H NMR(500MHz,DMSO-d6)δ9.39(d,J=10.7Hz,1H),9.20(d,J=11.4Hz,1H),8.70(d,J=8.8Hz,1H),8.01(s ,1H),7.74(d,J=2.1Hz,1H),7.69(d,J=8.3Hz,1H),7.61(s,1H),7.56(dd,J=8.3,2.1Hz,1H),4.71–4.6 3(m,2H),4.62–4.52(m,2H),4.47(tdd,J=11.7,8.6,4.3Hz,1H),3.40–3.35(m,2H),3.23(td,J=11.0, 5.8Hz, 1H), 2.94 (dd, J=24.0, 11.9Hz, 2H), 2.05 (td, J=10.4, 9.4, 3.4Hz, 2H). ESI-MS: m / z=575.0[M+H] + .

[0319] Example 21. Synthesis of 10-bromo-N-(((3S,4S)-4-(3-chloro-4-trifluoromethylphenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 10)

[0320]

[0321] Synthesis steps: Referring to Example 13, intermediate 17 and intermediate 2 were used as raw materials to prepare compound 10 with a yield of 51%. 1 H NMR(500MHz,DMSO-d6)δ8.07(s,1H),7.70(d,J=8.0Hz,1H),7.60(s,1H),7.4 5(d,J=5.8Hz,2H),4.63(dd,J=6.3,3.3Hz,2H),4.59–4.46(m,3H),3.65–3.59 (m,1H),3.59–3.52(m,1H),3.48(q,J=7.0Hz,1H),3.19(dt,J=24.1,12.4Hz, 2H),2.21(d,J=14.4Hz,1H),2.12(t,J=13.1Hz,1H).ESI-MS:m / z=575.0[M+H] + .

[0322] Example 22. Synthesis of 10-bromo-N-(((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 11)

[0323]

[0324] Synthesis steps: Referring to Example 13, intermediate 18 and intermediate 2 were used as raw materials to prepare compound 11 with a yield of 56%. 1 H NMR(500MHz,DMSO-d6)δ9.39(d,J=10.9Hz,1H),9.24(d,J=11.4Hz,1H),8.75(d ,J=8.9Hz,1H),8.08(s,1H),7.61(s,1H),7.21(dd,J=9.1,6.5Hz,2H),4.69–4. 54(m,4H),4.39(ddt,J=16.2,11.4,5.8Hz,1H),3.41–3.33(m,2H),3.16(td,J= 11.5,4.4Hz,1H),2.98–2.89(m,2H),2.09–1.96(m,2H).ESI-MS:m / z=527[M+H] + .

[0325] Example 23. Synthesis of 10-chloro-N-(((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 12)

[0326]

[0327] Synthesis steps: Referring to Example 13, intermediate 18 and intermediate 3 were used as raw materials to prepare compound 12 with a yield of 49%. 1 H NMR(500MHz,DMSO-d6)δ9.51(d,J=10.7Hz,1H),9.37(d,J=11.1Hz,1H),8.86(d, J=8.8Hz,1H),8.06(s,1H),7.61(s,1H),7.22(dd,J=9.1,6.6Hz,2H),4.67(dt,J =4.8,2.3Hz,2H),4.64–4.54(m,2H),4.45–4.36(m,1H),3.41–3.33(m,2H),3.20 (td,J=11.7,4.0Hz,1H),2.94(s,2H),2.13–1.98(m,2H).ESI-MS:m / z=483[M+H] + .

[0328] Example 24. Synthesis of 1-bromo-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolino[1,5-a]thiophene[3,2-c]azepane-9-carboxamide (Compound 13)

[0329]

[0330] Synthesis steps: Referring to Example 13, intermediate 15 and intermediate 2 were used as raw materials to prepare compound 13 with a yield of 77.2%. 1 H NMR (500MHz, DMSO-d6) δ9.48 (s, 1H), 9.40 (s, 1H), 8.79 (d, J = 8.8Hz, 1H), 8.03 (s, 1H) ),7.61(s,1H),7.44–7.24(m,2H),7.12(s,1H),4.54–4.41(m,1H),4.31–4.19(m,J=4 .4Hz,2H),3.38(d,J=11.9Hz,2H),3.21–3.10(m,1H),3.02(t,J=6.9Hz,2H),2.93(q, J=12.5,12.1Hz,2H),2.31–2.16(m,2H),2.11–1.99(m,2H).ESI-MS:m / z=507.1[M+H] + .

[0331] Example 25. Synthesis of 1-bromo-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolino[1,5-a]thiophene[3,2-c]azepane-9-carboxamide (Compound 14)

[0332]

[0333] Synthesis steps: Referring to Example 13, intermediate 13 and intermediate 5 were used as raw materials to prepare compound 14 with a yield of 86.1%. 1 H NMR(500MHz,DMSO-d6)δ9.48(d,J=8.8Hz,1H),9.38(d,J=10.0Hz,1H),8.81(d,J=8.9Hz,1H),8.0 4(s,1H),7.60(s,1H),7.06(ddd,J=9.3,7.2,2.1Hz,1H),6.98(d,J=6.6Hz,2H),4.47(qd,J=11.6, 4.4Hz,1H),4.31–4.16(m,2H),3.36(d,J=11.4Hz,2H),3.19(td,J=11.6,4.2Hz,1H),3.01(t,J=7 .0Hz,2H),2.92(q,J=10.9Hz,2H),2.28–2.14(m,2H),2.11–1.98(m,2H).ESI-MS:m / z=507.1[M+H] + .

[0334] Example 26. Synthesis of 1-chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolino[1,5-a]thiophene[3,2-c]azepane-9-carboxamide (Compound 15)

[0335]

[0336] Synthesis steps: Referring to Example 13, intermediate 12 and intermediate 6 were used as raw materials to prepare compound 15 with a yield of 77.7%; 1H NMR(400MHz,Chloroform-d)δ7.66(s,1H),7.38(s,1H),7.07(td,J=10.1,9.0,3.7Hz,2H),6.99(d d,J=8.1,5.3Hz,1H),4.22(dd,J=7.4,4.2Hz,2H),4.20–4.12(m,1H),3.52(dd,J=12.2,4.3Hz,1H) ,3.19(d,J=13.0Hz,1H),3.06(t,J=6.9Hz,2H),2.75(td,J=11.9,11.5,3.5Hz,2H),2.70–2.58(m, 1H),2.26–2.23(m,2H),1.94(dd,J=13.6,3.5Hz,1H),1.81–1.70(m,1H).ESI-MS:m / z=463.0[M+H] + .

[0337] Example 27. Synthesis of N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolino[1,5-a]thiophene[3,2-c]azepane-9-carboxamide (Compound 16)

[0338]

[0339] Synthesis steps: Referring to Example 13, intermediate 12 and intermediate 4 were used as raw materials to prepare compound 16 with a yield of 86.6%; 1 H NMR(400MHz,Chloroform-d)δ7.33(d,J=18.2Hz,2H),7.06(q,J=8.6Hz,2H),6.98(t,J=6.2Hz,1H), 6.26(s,1H),6.03(d,J=8.2Hz,1H),4.49–4.27(m,2H),4.20(dd,J=11.3,7.1Hz,1H),3.47(dd,J=12. 2,4.3Hz,1H),3.15(d,J=11.7Hz,1H),3.11(t,J=6.3Hz,2H),2.71(t,J=12.1Hz,2H),2.59(t,J=11. 2Hz,1H),2.19(p,J=5.6Hz,2H),1.94(s,1H),1.68(tt,J=12.3,6.2Hz,1H).ESI-MS:m / z=429.0[M+H] + .

[0340] Example 28. Synthesis of 1-bromo-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolino[1,5-a]thiophene[3,2-c]azepane-9-carboxamide (Compound 17)

[0341]

[0342] Synthesis steps: Referring to Example 13, intermediate 14 and intermediate 5 were used as raw materials to prepare compound 17 with a yield of 89.3%; 1 H NMR(500MHz,DMSO-d6)δ9.46(d,J=8.6Hz,1H),9.35(d,J=10.7Hz,1H),8.74(d,J=8.9Hz,1H),8.01(s ,1H),7.60(s,1H),7.32(q,J=7.8Hz,1H),7.09(dd,J=17.0,9.0Hz,2H),7.01(td,J=8.6,2.2Hz,1H),4 .49(qd,J=11.6,4.4Hz,1H),4.35–4.13(m,2H),3.39–3.31(m,2H),3.14(td,J=11.6,4.1Hz,1H),3.0 1(t,J=7.0Hz,2H),2.97–2.84(m,2H),2.27–2.12(m,2H),2.10–1.99(m,2H).ESI-MS:m / z=489.0[M+H] + .

[0343] Example 29. Synthesis of 1-chloro-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolino[1,5-a]thiophene[3,2-c]azepane-9-carboxamide (Compound 18)

[0344]

[0345] Synthesis steps: Referring to Example 13, intermediate 14 and intermediate 6 were used as raw materials to prepare compound 18 with a yield of 92.2%; 1H NMR (500MHz, DMSO-d6) δ9.52(s,1H),9.44(d,J=10.0Hz,1H),8.78(d,J=8.9Hz,1H),8.02(s,1H),7.58( s,1H),7.31(q,J=7.8Hz,1H),7.09(dd,J=14.2,9.0Hz,2H),7.03–6.96(m,1H),4.48(qd,J=11.6,4.4Hz ,1H),4.25(p,J=10.3Hz,2H),3.35(d,J=11.9Hz,2H),3.15(td,J=11.8,3.6Hz,1H),3.06–3.04(m,2H), 2.92(q,J=11.2Hz,2H),2.16(dp,J=13.5,7.5,7.0Hz,2H),2.09–1.94(m,2H).ESI-MS:m / z=445.1[M+H] + .

[0346] Example 30. Synthesis of N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolino[1,5-a]thiophene[3,2-c]azepane-9-carboxamide (Compound 19)

[0347]

[0348] Synthesis steps: Referring to Example 13, intermediate 14 and intermediate 4 were used as raw materials to prepare compound 19 with a yield of 78.6%; 1 H NMR(500MHz,DMSO-d6)δ9.45(d,J=9.3Hz,1H),9.36(d,J=9.9Hz,1H),9.03(d,J=8.9Hz,1H),8.25(s,1H),7.41 (d,J=1.7Hz,1H),7.30(dq,J=14.1,7.9,6.2Hz,1H),7.18–7.08(m,2H),6.98(td,J=8.7,2.2Hz,1H),6.59(d,J= 1.6Hz,1H),4.49(qd,J=11.8,4.6Hz,1H),4.44–4.37(m,2H),3.35(d,J=11.7Hz,2H),3.23(td,J=11.6,4.1Hz, 1H),3.17–3.08(m,2H),2.96(q,J=10.9Hz,2H),2.17–2.08(m,2H),2.06–1.95(m,2H).ESI-MS:m / z=411.1[M+H] + .

[0349] Step 31. Synthesis of 1-chloro-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (Compound 20)

[0350]

[0351] Synthesis steps: Referring to Example 13, intermediate 13 and intermediate 7 were used as raw materials to prepare compound 20 with a yield of 88.2%; 1 H NMR (500MHz, DMSO) δ9.47(d,J=9.6Hz,1H),9.38(d,J=11.6Hz,1H),8.83(d,J=8.9Hz,1H),7.78(d,J=8.1Hz,1H),7 .72(s,1H),7.59(dd,J=8.2,1.9Hz,1H),7.51(d,J=1.8Hz,1H),7.05(ddd,J=9.3,6.9,2.4Hz,1H),7.04–6.95(m,2H ),4.59(ddt,J=14.8,9.8,5.0Hz,1H),4.53(q,J=4.5,3.8Hz,2H),4.46(dd,J=6.4,4.7Hz,2H),3.38(t,J=3.5Hz,2 H),3.22(td,J=11.3,4.8Hz,1H),2.95(q,J=11.1Hz,2H),2.12–2.04(m,1H),2.03(s,1H).ESI-MS:m / z=459.1[M+H] + .

[0352] Example 32. Synthesis of N-(((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (Compound 21)

[0353]

[0354] Synthesis steps: Referring to Example 13, intermediate 12 and intermediate 7 were used as raw materials to prepare compound 21 with a yield of 89.0%; 1H NMR(500MHz,MeOD)δ8.01–7.93(m,2H),7.41(dd,J=8.2,1.6Hz,1H),7.35(d,J=1.6Hz,1H),7.33– 7.26(m,1H),7.20–7.15(m,2H),7.14(d,J=2.4Hz,1H),4.83–4.75(m,2H),4.63(td,J=11.3,3.9Hz ,1H),4.57–4.52(m,2H),3.61(dd,J=12.1,4.3Hz,1H),3.55(d,J=12.2Hz,1H),3.27–3.20(m,2H) ,3.17(d,J=11.9Hz,1H),2.19(d,J=14.3Hz,1H),2.11(d,J=12.7Hz,1H).ESI-MS:m / z=425.1[M+H] +

[0355] Example 33. Synthesis of N-(((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (Compound 21)

[0356]

[0357] Synthesis steps: Referring to Example 13, intermediate 14 and intermediate 7 were used as raw materials to prepare compound 22 with a yield of 94.5%; 1 H NMR (500MHz, DMSO) δ9.56(d,J=10.7Hz,1H),9.46(q,J=11.0Hz,1H),8.70(d,J=8.8Hz,1H),7.89(d,J=8.4Hz,1H),7.51(d,J =1.9Hz,1H),7.40(dd,J=8.3,1.8Hz,1H),7.35(d,J=1.7Hz,1H),7.30(p,J=8.7,8.0Hz,1H),7.15–7.04(m,2H),6.99(td,J= 8.6,2.5Hz,1H),6.93(d,J=2.0Hz,1H),4.64–4.59(m,2H),4.59–4.54(m,1H),4.43(s,2H),3.36(dq,J=11.8,5.9,4.6Hz,2H ),3.19(td,J=11.7,3.9Hz,1H),2.94(p,J=11.2Hz,2H),2.12–2.02(m,1H),1.99(d,J=14.8Hz,1H).ESI-MS:m / z=407.1[M+H] + .

[0358] Example 34. Synthesis of N-(((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (Compound 23)

[0359]

[0360] Synthesis steps: Referring to Example 13, intermediate 14 and intermediate 7 were used as raw materials to prepare compound 23 with a yield of 68.2%; 1 H NMR(500MHz,MeOD)δ7.93(d,J=8.4Hz,1H),7.91–7.86(m,1H),7.40(dd,J=8.3,1.8Hz,1H),7.35(d,J=1.7Hz ,1H),7.07(d,J=2.5Hz,1H),7.00(h,J=4.3Hz,2H),6.80(tt,J=9.0,2.3Hz,1H),4.77–4.71(m,2H),4.64(td, J=11.5,4.2Hz,1H),4.55–4.48(m,2H),3.61(dd,J=12.3,4.4Hz,1H),3.58–3.52(m,1H),3.25(ddd,J=24.1, 12.5,9.1Hz,2H),3.16(d,J=12.0Hz,1H),2.21(d,J=14.1Hz,1H),2.17–2.04(m,1H)ESI-MS:m / z=425.1[M+H] + .

[0361] Example 35. Synthesis of N-(((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (Compound 24)

[0362]

[0363] Synthesis steps: Referring to Example 13, intermediate 18 and intermediate 7 were used as raw materials to prepare compound 24 with a yield of 68.2%; 1H NMR(500MHz,DMSO-d6)δ9.39(d,J=10.9Hz,1H),9.24(d,J=11.4Hz,1H),8.75(d ,J=8.9Hz,1H),8.08(s,1H),7.61(s,1H),7.21(dd,J=9.1,6.5Hz,2H),4.69–4. 54(m,4H),4.39(ddt,J=16.2,11.4,5.8Hz,1H),3.41–3.33(m,2H),3.16(td,J= 11.5,4.4Hz,1H),2.98–2.89(m,2H),2.09–1.96(m,2H).ESI-MS:m / z=443[M+H] + .

[0364] Example 36. Synthesis of 10-bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(dimethylamino)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 25)

[0365]

[0366] Intermediate 22 (mg, mmol) was dissolved in 3 ml of DMF solution, and N,N-dimethylamine (mg, mmol) was added. The temperature was raised to 90°C and stirred for 2 h. After the reaction was completed as monitored by TLC, the mixture was cooled to room temperature and poured into 15 ml of water. The mixture was extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a pale yellow solid (mg, mmol). The yield was %. The pale yellow solid (mg, mmol) was dissolved in 3 ml of 4N HCl-1,4-dioxane solution and stirred at room temperature for 1 h. The reaction was then analyzed by TLC. After the reaction was completed, the mixture was dried by spin drying. Saturated sodium bicarbonate was added and the mixture was extracted twice with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain a white solid product (Compound 25). The yield was %; ESI-MS: m / z=594 [M+H] + .

[0367] Example 37. Synthesis of N-((3S,4S,6R)-6-allyl-4-(3,4-difluorophenyl)piperidin-3-yl)-10-bromo-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 26)

[0368]

[0369] Synthesis steps: Referring to Example 12, compound 26 was prepared from compound 7-2 with a yield of 58%. 1 H NMR(500MHz,Methanol-d4)δ8.11(s,1H),7.47(s,1H),7.28–7.18(m,2H),7.12(s,1H),5.85(dd t,J=17.1,10.2,7.0Hz,1H),5.40(dd,J=17.0,1.5Hz,1H),5.30(dd,J=10.1,1.5Hz,1H),4.76–4. 63(m,2H),4.63–4.51(m,2H),4.44(td,J=11.4,4.7Hz,1H),3.79(s,1H),3.40(td,J=12.3,4.9H z,2H),2.81–2.68(m,2H),2.12(dd,J=5.0,2.4Hz,1H),1.32–1.27(m,2H).ESI-MS:m / z=549[M+H] + .

[0370] Example 38. Synthesis of N-((3S,4S,6R)-6-(3-(1H-imidazol-1-yl)propyl)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-bromo-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 27)

[0371]

[0372] Synthesis steps: Referring to Example 34, intermediate 22 and imidazole were used as raw materials to prepare compound 27. Yield: % ESI-MS: m / z = 617 [M+H] + .

[0373] Example 39 Synthesis of 10-bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-hydroxypropyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 28)

[0374]

[0375] Synthesis steps: Referring to Example 12, compound 28 was prepared from compound 7-3. Yield: % ESI-MS: m / z=567 [M+H] +

[0376] Example 40 Synthesis of 10-bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(pyrrolidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 29)

[0377]

[0378] Synthesis steps: Referring to Example 34, intermediate 22 and pyrrole were used as raw materials to prepare compound 29 with a yield of %. 1 HNMR(500MHz,Methanol-d4)δ8.14(s,1H),7.47(s,1H),7.34(t,J=9.2Hz,1H),7.18(d,J=13.1Hz ,2H),4.65(s,2H),4.60–4.50(m,2H),4.45(d,J=10.9Hz,1H),3.78–3.74(m,1H),3.73–3.70(m,1H ),3.58–3.54(m,1H),3.40(d,J=7.9Hz,2H),3.27–3.23(m,2H),3.15(s,1H),2.75(s,2H),2.24–2. 19(m,2H),2.18(s,2H),2.06(s,2H),2.02–1.99(m,2H),1.95–1.88(m,2H).ESI-MS:m / z=620[M+H] +

[0379] Example 41. Synthesis of 10-bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(piperidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 30)

[0380]

[0381] Synthesis steps: Referring to Example 34, intermediate 22 and piperidine were used as raw materials to prepare compound 30. Yield: % ESI-MS: m / z = 634 [M+H] +

[0382] Example 42. Synthesis of 10-bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-morpholinopropyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 31)

[0383]

[0384] Synthesis steps: Referring to Example 34, intermediate 22 and morpholine were used as raw materials to prepare compound 31. Yield: % ESI-MS: m / z=636[M+H]+

[0385] Example 43. Synthesis of 10-bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(4-hydroxypiperidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 32)

[0386]

[0387] Synthesis steps: Referring to Example 34, intermediate 22 and 4-hydroxypiperidine were used as raw materials to prepare compound 32. Yield: % ESI-MS: m / z = 650 [M+H] +

[0388] Example 44. Synthesis of 10-bromo-N-(((3S,4S,6S)-4-(3,4-difluorophenyl)-6-(2-(methylamino)-2-oxoethyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 33)

[0389]

[0390] Synthesis steps: Referring to Example 13, intermediate 22 and intermediate 2 were used as raw materials to prepare compound 33. Yield: % ESI-MS: m / z = 580 [M+H] +

[0391] Example 45. Synthesis of 10-chloro-N-(((3S,4S,6S)-4-(3,4-difluorophenyl)-6-(2-(methylamino)-2-oxoethyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 34)

[0392]

[0393] Synthesis steps: Referring to Example 13, intermediate 22 and intermediate 3 were used as raw materials to prepare compound 34 with a yield of 66%. 1H NMR(500MHz,Methanol-d4)δ8.06(s,1H),7.46(s,1H),7.33–7.09(m,3H),4.64(dq,J=4.4,2.1Hz,2 H),4.57(dq,J=5.1,2.1Hz,2H),4.41(td,J=11.5,4.9Hz,1H),4.09(dd,J=9.8,5.1Hz,1H),3.44(dd ,J=13.0,4.9Hz,1H),3.40–3.33(m,2H),3.02(dd,J=16.2,9.9Hz,1H),2.78(s,3H),2.76–2.71(m,1 H),2.23(ddd,J=14.8,12.9,5.1Hz,1H),2.05(ddd,J=14.8,3.9,1.8Hz,1H).ESI-MS:m / z=536[M+H] +

[0394] Example 46. Synthesis of N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-ethyl-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 35)

[0395]

[0396] Step 1 Synthesis of compound 11-1

[0397] The synthesis steps were as shown in step 2 of Example 5. Compound 11-1 was prepared using intermediate 6-1 as a raw material with a yield of 80.6%.

[0398] Step 2 Synthesis of compound 35

[0399] The synthesis steps were as shown in Example 12. Compound 35 was prepared using compound 11-1 as the raw material with a yield of 43.2%. 1HNMR(400MHz,DMSO-d6)δ9.50(s,1H),9.41(s,1H),9.01(d,J=10.4Hz,1H),7.78–7.71(m,1H ),7.37–7.29(m,3H),7.15–7.09(m,1H),4.57(t,J=4.4Hz,2H),4.52–4.49(m,2H),4.40(d,J= 7.5Hz,1H),3.39–3.31(m,2H),3.25(t,J=11.8Hz,1H),2.97(dt,J=25.3,12.1Hz,2H),2.68(q d,J=7.6,2.6Hz,2H),2.08(q,J=12.8,11.9Hz,1H),2.02–1.92(m,1H),1.16(t,J=7.4Hz,3H).

[0400] Example 47. Synthesis of N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-(2-hydroxyethyl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 36)

[0401]

[0402] Step 1 Synthesis of Compound 12-1

[0403] The synthesis steps were as follows: Refer to step 3 of Example 7, and use intermediate 25 as the raw material to prepare compound 12-1 with a yield of .

[0404] Step 2 Synthesis of compound 36

[0405] The synthesis steps were as described in Example 12. Compound 36 was prepared using compound 12-1 as the starting material with a yield of 45%. 1H NMR(500MHz,DMSO-d6)δ9.15(d,J=10.9Hz,1H),9.05(d,J=9.1Hz,2H),7.74(s,1H),7.37–7.28 (m,3H),7.14(dt,J=9.0,2.9Hz,1H),4.59–4.54(m,2H),4.53–4.46(m,2H),4.38(dtd,J=11.2,8 .0,7.1,4.4Hz,1H),3.58(s,1H),3.36–3.33(m,1H),3.27(td,J=8.9,8.4,4.7Hz,1H),3.06–2. 90(m,2H),2.89(s,1H),2.82(dq,J=18.6,7.2Hz,2H),2.73(s,1H),2.00(dd,J=9.2,3.4Hz,2H).

[0406] Example 48. Synthesis of 10-bromo-N-(((3R,4R)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (Compound 37)

[0407]

[0408] Synthesis steps: Referring to Example 13, intermediate 19 and intermediate 2 were used as raw materials to prepare compound 37 with a yield of %. 1 HNMR(500MHz,DMSO-d6)δ9.46(s,1H),9.38–9.23(m,1H),8.68(dt,J=9.0,2.2Hz,1H),8.03(d,J=1 .4Hz,1H),7.60(s,1H),7.36(dt,J=10.8,8.5Hz,1H),7.29(ddd,J=12.1,7.8,2.1Hz,1H),7.15–7.0 6(m,1H),4.73–4.65(m,2H),4.62–4.51(m,2H),4.39(ddd,J=7.0,4.7,2.4Hz,1H),3.43–3.34(m,2 H),3.11(td,J=11.6,4.4Hz,1H),3.00–2.85(m,2H),1.99(d,J=11.0Hz,2H).ESI-MS:m / z=509[M+H] +

[0409] Example 49. Synthesis of N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide (Compound 38)

[0410]

[0411] Synthesis steps: Referring to Example 13, intermediate 9 and intermediate 2 were used as raw materials to prepare compound 38. Yield: % ESI-MS: m / z = 444 [M+H] +

[0412] Example 50. Synthesis of 10-chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide (Compound 39)

[0413]

[0414] Synthesis steps: Referring to Example 13, intermediates 11 and 12 were used as raw materials to prepare compound 39. Yield: % ESI-MS: m / z = 479 [M+H] +

[0415] Example 51. Synthesis of 10-bromo-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide (Compound 40)

[0416]

[0417] Synthesis steps with reference to Example 13, using intermediates 10 and 12 as raw materials, to prepare compound 40, yield%. 1H NMR(400MHz,Chloroform-d)δ7.86(s,1H),7.51(s,1H),7.19(dt,J=7.7,5.4Hz,1 H),7.09(tdd,J=9.4,5.6,3.0Hz,2H),4.85(d,J=11.2Hz,2H),4.45(dd,J=10.5,6 .3Hz,1H),3.55(d,J=4.6Hz,1H),3.46(d,J=12.6Hz,1H),3.10(dtd,J=11.8,8.3, 7.3,3.7Hz,3H),2.15–2.11(m,1H),1.98(d,J=12.9Hz,1H).ESI-MS:m / z=522[M+H] +

[0418] Example 52 Synthesis of N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-methyl-5,6-dihydropyrazolo[1,5-d]thiophene[3,2-f][1,4]oxazepine-2-carboxamide (Compound 41)

[0419]

[0420] Synthesis steps: Referring to Example 13, intermediate 12 and intermediate 25 were used as raw materials to prepare compound 39. Yield: % ESI-MS: m / z = 479 [M+H] +

[0421] The compound disclosed in the present invention has growth inhibition and differentiation promotion effects on neuroblasts, as well as AKT1 kinase inhibitory activity.

[0422] Using the clinical differentiation promoter 13-cisRA as a positive control, the cell synapse elongation change phenotypic screening method was used to evaluate the differentiation ratio, average synapse length and maximum synapse length of common tumor cell lines (neuroblast cell lines). The MTT method was used to determine the in vitro inhibitory effect (IC50) of the compounds on them. At the same time, a commercial AKT1 kit was used to evaluate the AKT1 enzyme inhibitory activity (IC50).

[0423] The pharmacological experimental methods and results of the anti-tumor activity of the compounds of the present invention are as follows:

[0424] The first step was to determine the in vitro tumor-inducing differentiation activity and conduct a preliminary structure-activity relationship study. Neuroblastoma cell lines were used to determine the in vitro tumor-inducing differentiation activity of the synthesized compounds.

[0425] Experimental Materials

[0426] Cell line: Mouse neuroblastoma cells (Neuro2a)

[0427] Culture medium: RPMI1640 + fetal bovine serum

[0428] Drug preparation method: Dissolve the drug in DMSO to prepare a 50mM stock solution, and dilute it in a certain proportion to obtain 5 different concentrations

[0429] Tumor cell culture in vitro:

[0430] The selected Neuro2a cells were incubated in a 37°C, 5% CO2 cell culture incubator and passaged (adherent cells were digested with Duck's EDTA before passage) when the cell density reached 70-90% for future experiments.

[0431] Appropriate numbers of NB Neruo2a cells were seeded into 6-well plates and cultured overnight in a cell culture incubator. After the cells attached and grew, they were treated with various compounds at a final concentration of 0.5 μM, with DMSO (1%) serving as a blank control. After 48 hours of treatment, images were taken. Changes in synaptic elongation were calculated using the NeuonJ plugin in ImageJ software. The experiment was repeated three times. Differentiation strength was assessed using the differentiation ratio (number of synapses / number of cells), average synaptic length, and maximum synaptic length. All data were normalized by dividing by the control group.

[0432] The calculation formula for differentiation ratio is: differentiation ratio = (number of axonal cells in the experimental group / total number of cells in the experimental group) / (number of axonal cells in the control group / total number of cells in the control group); average neurite length = (average neurite length in the experimental group) / (average neurite length in the control group); maximum neurite length = (maximum neurite length in the experimental group) / (maximum neurite length in the control group).

[0433] The second is the test method for the inhibitory activity of the test compound on Neuro2a cell proliferation:

[0434] Cells in the logarithmic growth phase were seeded in 96-well culture plates at a density of 4 × 103 cells / well. Complete medium supplemented with 10% serum was added to each well and the cells were cultured overnight in a 20% oxygen incubator. After the cells adhered, a gradient of test compounds was added and the cells were cultured in a normoxic incubator for an additional 3 days to evaluate the inhibitory effect of the candidate compounds on cell proliferation. The OD value of each well was measured using a microplate reader (detection wavelength: 490 nm), and the obtained data were used to calculate the IC50 value.

[0435] The calculation formula of cell inhibition rate is: cell inhibition rate (%) = (OD value of control group - OD value of drug group) / OD value of control group × 100%. The half inhibition concentration IC50 value was calculated by Bliss method and obtained by Graphpad5.0 software.

[0436] Finally, the test method for the inhibition rate of the test compound on AKT1 enzyme:

[0437] Mobility shift assay was used to detect the inhibitory activity of compounds on AKT1.

[0438] The compound was dissolved in 100% DMSO to a 10 mM stock solution, which was stored in a nitrogen cabinet protected from light. Preparation of a compound concentration gradient: The starting concentration of the test compound was 1000 nM, and six concentrations were tested in a single well using 10-fold or 3-fold dilutions. The solution was diluted to 100 times the final concentration in a 384-well plate. Then, 250 nl was transferred to a 384-well reaction plate using an Echo550 for later use. 250 nl of 100% DMSO was added to each of the negative and positive control wells. 10 μl of kinase solution at 2.5 times the final concentration was added to each of the compound and positive control wells; 10 μl of 1× Kinase buffer was added to the negative control well. Centrifuge at 1000 rpm for 30 seconds, shake the reaction plate to mix, and incubate at room temperature for 10 minutes. A mixed solution of ATP and Kinase substrate 6 at 25 / 15 times the final concentration was prepared using 1× Kinase buffer. Initiate the reaction by adding 15 μl of a mixture of ATP and substrate at 25 / 15 times the final concentration. Centrifuge the 384-well plate at 1000 rpm for 30 seconds, vortex to mix, and incubate at room temperature for 30 minutes. Stop the kinase reaction by adding 30 μl of stop assay solution, centrifuge at 1000 rpm for 30 seconds, and vortex to mix. Read the conversion rate using a Caliper EZ Reader.

[0439] The inhibition rate was calculated as follows: Inhibition% = (Conversion%_max - Conversion%_sample) / (Conversion%_max - Conversion%_min) * 100, where Conversion%_sample is the conversion rate reading of the sample; Conversion%_min is the mean of the negative control wells, representing the conversion rate reading of the wells without enzyme activity; and Conversion%_max is the mean of the positive control wells, representing the conversion rate reading of the wells without compound inhibition.

[0440] IC50 was calculated as follows: Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope)). The logarithm of the concentration was used as the X-axis, and the log(inhibitor) vs. response-variable slope function in GraphPad Prism 5 was used to fit the dose-effect curve to determine the IC50 value for each compound on enzyme activity.

[0441] Table 1 IC50 (μM) of some compounds on tumor cell proliferation and Akt1 kinase

[0442]

[0443]

[0444] Table 2 Differentiation-promoting activity of some compounds on Neuro2a

[0445]

[0446]

[0447] a.Diff rate=Differentiation rate: the number of axons / cells; normalized to control

[0448] b.Avg neurite=Average neurite length; normalized to control

[0449] c.Max neurite=maximum neurite length; normalized to control

[0450] Figure 1 This is the differentiation of Neuro2a cells in the non-medicated group. Figure 2 The results are as follows: The differentiation of Neuro2a cells in the drug group (Compound 1). By comparison, it can be found from the phenotype that Compound 1 can significantly promote the synaptic growth of Neuro2a cells. This shows that the compounds of the present invention represented by Compound 1 have the ability to promote the differentiation of Neuro2a cells.

Claims

1. Compounds represented by general formula (IV) and (IV'): ； and pharmaceutically acceptable salts thereof, wherein: R1 is selected from unsubstituted or substituted phenyl; R2 is selected from H, allyl, 、 、 、 、 , 3-(piperidin-1-yl)propyl, 3-morpholinopropyl, 3-(4-hydroxypiperidin-1-yl)propyl, wherein: n = an integer of 0-4; R d is selected from H, C1~C4 alkyl, halogenated C1~C4 alkyl; Re is selected from C1~C4 alkyl, halogenated C1~C4 alkyl; Ring D is selected from unsubstituted or substituted five-membered saturated aliphatic nitrogen-containing heterocyclic group, unsubstituted or substituted five-membered nitrogen-containing heteroaryl group, and the carbon atoms in the heterocyclic group and heteroaryl group may be further substituted by O or S; R3 is selected from H, halogen, hydroxyl, hydroxymethyl, hydroxyethyl, carboxyl, saturated or unsaturated C1~C4 hydrocarbon group, halogenated C1~C4 alkyl, C1~C4 alkoxy, halogenated C1~C4 alkoxy; R4 is selected from H, halogen, nitro, amino, cyano, C1~C4 alkyl, halogenated C1~C4 alkyl, C1~C4 alkoxy, halogenated C1~C4 alkoxy; X is selected from O, Y is selected from CH2; X is selected from CH2, Y is selected from CH2; X is selected from NH; Y is selected from C(=O); E is S; The substituent is selected from halogen, nitro, amino, cyano, hydroxy, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, and halogenated C1-C3 alkoxy.

2. The compound according to claim 1, characterized in that X is O or CH2; Y is CH2; The R1 is selected from substituted phenyl; the substituent is one or more halogen, halogenated C1~C3 alkyl; R2 is selected from H, 3-(dimethylamino)propyl, allyl, 3-(1H-imidazol-1-yl)propyl, 3-hydroxypropyl, 3-(pyrrolidin-1-yl)propyl, 3-(piperidin-1-yl)propyl, 3-morpholinopropyl, 3-(4-hydroxypiperidin-1-yl)propyl, 2-(methylamino)-2-oxoethyl; R3 is selected from H, halogen, C1~C5 alkyl; E stands for S.

3. The compound according to claim 1, characterized in that The compound is selected from: 10-Bromo-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (1) 10-Chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (2) N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (3) 10-Chloro-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (4) 10-Bromo-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (5) N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (6) 10-Bromo-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolino[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (7) 10-Bromo-N-((3S,4S)-4-(4-chlorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (8) 10-Bromo-N-((3S,4S)-4-(4-chloro-3-trifluoromethylphenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (9) 10-Bromo-N-((3S,4S)-4-(3-chloro-4-(trifluoromethyl)phenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (10) 10-Bromo-N-((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (11) 10-Chloro-N-((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (12) 1-Bromo-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thieno[3,2-c]azepane-9-carboxamide (13) 1-Bromo-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thieno[3,2-c]azepane-9-carboxamide (14) 1-Chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thienyl[3,2-c]azepane-9-carboxamide (15) N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thieno[3,2-c]azepane-9-carboxamide (16) 1-Bromo-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thieno[3,2-c]azepane-9-carboxamide (17) 1-Chloro-N-((3S,4S)-4-(3-fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thieno[3,2-c]azepane-9-carboxamide (18) N-((3S,4S)-4-(3-Fluorophenyl)piperidin-3-yl)-6,7-dihydro-5H-pyrazolo[1,5-a]thieno[3,2-c]azepane-9-carboxamide (19) 1-Chloro-N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (20) N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (21) N-((3S,4S)-4-(3-Fluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (22) N-((3S,4S)-4-(3,5-difluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (23) N-((3S,4S)-4-(3,4,5-trifluorophenyl)piperidin-3-yl)-5,6-dihydrobenzo[f]pyrazolo[1,5-d][1,4]oxazepine-9-carboxamide (24) 10-Bromo-N-((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(dimethylamino)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (25) N-((3S,4S,6R)-6-allyl-4-(3,4-difluorophenyl)piperidin-3-yl)-10-bromo-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (26) N-((3S,4S,6R)-6-(3-(1H-imidazol-1-yl)propyl)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-bromo-5, 6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (27) 10-Bromo-N-((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-hydroxypropyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (28) 10-Bromo-N-((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(pyrrolidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (29) 10-Bromo-N-((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(piperidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (30) 10-Bromo-N-(((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-morpholinopropyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (31) 10-Bromo-N-((3S,4S,6R)-4-(3,4-difluorophenyl)-6-(3-(4-hydroxypiperidin-1-yl)propyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (32) 10-Bromo-N-((3S,4S,6S)-4-(3,4-difluorophenyl)-6-(2-(methylamino)-2-oxoethyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (33) 10-Chloro-N-((3S,4S,6S)-4-(3,4-difluorophenyl)-6-(2-(methylamino)-2-oxoethyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (34) N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-ethyl-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (35) N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-(2-hydroxyethyl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (36) 10-Bromo-N-((3R,4R)-4-(3,4-difluorophenyl)piperidin-3-yl)-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (37) N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide (38) 10-Chloro-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide (39) 10-Bromo-N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-5-oxo-5,6-dihydro-4H-pyrazolo[1,5-d]thieno[3,2-f][1,4]diazepine-2-carboxamide (40) N-((3S,4S)-4-(3,4-difluorophenyl)piperidin-3-yl)-10-methyl-5,6-dihydropyrazolo[1,5-d]thieno[3,2-f][1,4]oxazepine-2-carboxamide (41) and pharmaceutically acceptable salts of the above compounds.

4. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises at least one active ingredient and one or more pharmaceutically acceptable carriers or excipients, wherein the active ingredient is selected from the compound according to any one of claims 1 to 3, or any one or more of the pharmaceutically acceptable salts of the compound.

5. A preparation prepared from the pharmaceutical composition according to claim 4, wherein the preparation is in the form of tablets, powders, granules, capsules, oral solutions and injections.

6. Use of the compound according to any one of claims 1 to 3 in the preparation of an anti-tumor drug, wherein the tumor is selected from neuroblastoma, breast cancer, sarcoma, lung cancer, prostate cancer, colon cancer, rectal cancer, kidney cancer, pancreatic cancer, blood cancer, glioma, head cancer, neck cancer, thyroid cancer, pancreatic cancer, liver cancer, ovarian cancer, vulvar cancer, cervical cancer, endometrial cancer, testicular cancer, bladder cancer, esophageal cancer, gastric cancer, nasopharyngeal cancer, cheek cancer, oral cancer, gastrointestinal stromal cancer, skin cancer, and multiple myeloma.

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