CN109071550B

CN109071550B - Fused imidazole derivative with IDO/TDO inhibitory activity and preparation method and application thereof

Info

Publication number: CN109071550B
Application number: CN201780023049.0A
Authority: CN
Inventors: 包如迪; 李元念; 刘福萍; 张静涛
Original assignee: Jiangsu Hansoh Pharmaceutical Group Co Ltd; Shanghai Hansoh Biomedical Co Ltd
Current assignee: Jiangsu Hansoh Pharmaceutical Group Co Ltd; Shanghai Hansoh Biomedical Co Ltd
Priority date: 2016-04-27
Filing date: 2017-04-06
Publication date: 2021-04-06
Anticipated expiration: 2037-04-06
Also published as: CN107312005B; CN109071550A; CN107312005A; WO2017185959A1

Abstract

A fused imidazole derivative with IDO/TDO inhibitory activity and having a structure shown in formula (I), and a preparation method and application thereof. The series of condensed imidazole derivatives have high inhibitory activity on IDO/TDO, can be widely applied to treatment or prevention of cancers or tumors, viral infection, depression, neurodegenerative diseases, trauma, age-related cataract, organ transplant rejection or autoimmune diseases, can also be used for inhibiting immunosuppression of patients, and is expected to be developed into a new generation of immunosuppressant.

Description

Fused imidazole derivative with IDO/TDO inhibitory activity and preparation method and application thereof

Technical Field

The invention belongs to the technical field of drug development, and particularly relates to a fused imidazole derivative with IDO/TDO (ion-doped doods/deoxyribose nucleic acid) inhibitory activity, and a preparation method and application thereof.

Background

Tryptophan (Trp) is an essential amino acid for the human body and is an essential nutrient for the human body to synthesize proteins, nicotinic acid and the neurotransmitter 5-hydroxytryptamine (serotonin). Tryptophan has two metabolic pathways in the body: the 5-hydroxytryptamine pathway and the kynurenine pathway. A small portion of tryptophan is converted to 5-hydroxytryptamine by tryptophan hydroxylase, and about 95% of tryptophan is converted to kynurenine by indoleamine 2, 3-dioxygenase (IDO) or tryptophan 2, 3-dioxygenase (TDO). Kynurenine also has two metabolic pathways, most of kynurenine generates 3-hydroxykynurenine (3-hydroxykynurenine) under the action of kynurenine hydroxylase (kynurenine 3-hydroxyylase), then the Kynurenine (KYNU) catalyzes and hydrolyzes the kynurenine to generate 3-hydroxyanthranilic acid, and finally active molecules such as quinolinic acid, pyridine carboxylic acid and Nicotinamide Adenine Dinucleotide (NAD) are generated through multi-level enzymatic reaction to participate in various physiological processes in vivo; the other path is that canine uroquinolinic acid (KYNA) is generated under the action of kynurenine aminotransferase (kynurenic acid transferase I and II, KAT I/1I).

Indoleamine 2, 3-dioxygenase (IDO) is the rate-limiting enzyme of the tryptophan/kynurenine pathway, and acts on indoleamine-containing substrates more broadly than TDO in mammalian IDO, which is widely present in cells of tissues other than liver, including astrocytes, microglia, macrophages, and vascular endothelial cells. TDO is almost expressed in the liver and has high selectivity to a substrate.

IDO can be induced by pro-inflammatory factors, where interferon γ is the strongest inducer, activating IDO during the stimulatory activation of high levels of interferon γ, promoting tryptophan metabolism.

The research on a pregnant mouse model shows that the syncytium trophoblast cells and antigen presenting cells at the interface of a mother fetus can synthesize the IDO, the dynamic change of the expression of the IDO is consistent with the formation of an embryo, and if the synthesis of the IDO is specifically blocked, the IDO can cause abortion of a mouse, which indicates that the IDO can prevent immunological rejection and is an immunoregulatory enzyme. IDO is associated with immune escape of tumor cells to monitor and attack them by the immune system. A plurality of tumor cells highly express IDO antigen, so that local T cell proliferation is inhibited, and the tumor cells are mediated to escape from the attack of an immune system. IDO is involved in regulating T cell responses, T cells are sensitive to tryptophan depletion, T cell proliferation stops at G1 when tryptophan concentration decreases, and IDO inhibits T cell activation by degrading tryptophan.

IDO activation is closely related to pathogenesis of various diseases, is an important target in the fields of related diseases such as tumors, Alzheimer's disease, Parkinson's disease, depression and the like, can liberate the defense system of an organism and help T cells to attack tumors better aiming at IDO/TDO inhibitors, so that the IDO/TDO inhibitors have the potential of treating a wide range of tumors, have wide application prospect, but do not come into the market so far, and therefore, the search and development of novel efficient IDO inhibitors have important theoretical significance and application value.

The development of indoleamine 2, 3-dioxygenase inhibitors is currently in the early stages of development, including Indoximod by NewLink, NLG-919(IDO/TDO bispecific), Epacadostat by Incyte (INCB 024360), and IDO or TDO inhibitors by BMS, Flexus, Iomet, itos, curad, etc.; the compounds of the examples of the invention have high inhibitory activity against indoleamine 2, 3-dioxygenase (IDO) and inhibitory activity against tryptophan 2, 3-dioxygenase (TDO) in enzymatic and cellular models, and have good exposure in PK animal models.

Disclosure of Invention

In order to solve the defects of the prior art, the inventor finds a fused imidazole derivative with a structure shown as a formula (I) in the research process, the series of compounds have high inhibitory activity on IDO/TDO, and can be used for treating diseases with pathological characteristics of tryptophan metabolic disorder mediated by IDO/TDO singly or in combination. Can be widely applied to the treatment or prevention of cancers or tumors, virus infection, depression, neurodegenerative diseases, wounds, age-related cataract, organ transplant rejection or autoimmune diseases, can also be used for inhibiting the immunosuppression of patients, and is expected to be developed into a new generation of immunosuppressive agents.

In one aspect, the present invention provides a compound having the following formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

wherein,

R₁、R₂each independently selected from hydrogen, deuterium, halogen, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, C_5-10Aryl, 5-10 membered heteroaryl, -C_0-8-S(O)_rR₅、-C_0-8-O-R₆、-C_0-8-C(O)OR₆、-C_0-8-C(O)R₆、-C_0-8-O-C(O)R₇、-C_0-8-NR₈R₉、-C_0-8-C(O)NR₈R₉、-N(R₈)-C(O)R₇or-N (R)₈)-C(O)OR₆，

Optionally further substituted by one or more groups selected from halogen, cyano, nitro, azido, optionally substituted or unsubstituted C_1-8Alkyl, optionally substituted or unsubstituted C_3-8Cycloalkyl, optionally substituted or unsubstituted 3-8 membered heterocyclic group, optionally substituted or unsubstituted C_5-10Aryl, optionally substituted or unsubstituted 5-10 membered heteroaryl, -C_0-8-S(O)_rR₅、-C_0-8-O-R₆、-C_0-8-C(O)OR₆、-C_0-8-C(O)R₆、-C_0-8-O-C(O)R₇、-C_0-8-NR₈R₉、-C_0-8-C(O)NR₈R₉、-N(R₈)-C(O)R₇or-N (R)₈)-C(O)OR₆Substituted with the substituent(s);

R、R₃、R₄each independently selected from hydrogen, deuterium, halogen, cyano, nitro, azido, optionally substituted or unsubstituted C_1-8Alkyl, optionally substituted or unsubstituted C_2-8Alkenyl, optionally substituted or unsubstituted C_2-8Alkynyl, optionally substituted or unsubstituted C_3-8Cycloalkyl, optionally substituted or unsubstituted 3-8 membered heterocyclic group, optionally substituted or unsubstituted C_5-10Aryl, optionally substituted or unsubstituted 5-10 membered heteroaryl, -C_0-8-S(O)_rR₅、-C_0-8-O-R₆、-C_0-8-C(O)OR₆、-C_0-8-C(O)R₆、-C_0-8-O-C(O)R₇、-C_0-8-NR₈R₉、-C_0-8-C(O)NR₈R₉、-N(R₈)-C(O)R₇or-N (R)₈)-C(O)OR₆；

R₅、R₆Each independently selected from hydrogen, deuterium, optionally substituted or unsubstituted C_1-8Alkyl, optionally substituted or unsubstituted C_2-8Alkenyl, optionally substituted or unsubstituted C_2-8Alkynyl, optionally substituted or unsubstituted C_3-8Cycloalkyl, optionally substituted or unsubstituted 3-8 membered heterocyclic group, optionally substituted or unsubstituted C_5-10Aryl, optionally substituted or unsubstituted 5-to 10-membered heteroaryl, optionally substituted or unsubstituted amino or optionally substituted or unsubstituted C_1-8An alkanoyl group;

R₇selected from hydrogen, deuterium, optionally substituted or unsubstituted C_1-8Alkyl, optionally substituted or unsubstituted C_2-8Alkenyl, optionally substituted or unsubstituted C_2-8Alkynyl, optionally substituted or unsubstituted C_1-8Alkoxy, optionally substituted or unsubstituted C_3-8Cycloalkyl, optionally substituted or unsubstituted C_3-8Cycloalkoxy, optionally substituted or unsubstituted 3-8 membered heterocyclic group, optionally substituted or unsubstituted 3-8 membered heterocyclic oxy, optionally substituted or unsubstituted C_5-10Aryl, optionally substituted or unsubstituted C_5-10Aryloxy, optionally substituted or unsubstituted 5-10 membered heteroaryl, optionally substituted or unsubstituted 5-10 membered heteroaryloxy, or optionally substituted or unsubstituted amino;

R₈、R₉each independently selected from hydrogen, deuterium, hydroxy, optionally substituted or unsubstituted C_1-8Alkyl, optionally substituted or unsubstituted C_2-8Alkenyl, optionally substituted or unsubstituted C_2-8Alkynyl, optionally substituted or unsubstituted C_3-8Cycloalkyl, optionally substituted or unsubstituted 3-8 membered heterocyclic group, optionally substituted or unsubstituted C_5-10Aryl, optionally substituted or unsubstituted 5-to 10-membered heteroaryl, optionally substituted or unsubstituted C_1-8Alkanoyl or optionally substituted or unsubstituted amino;

the optionally substituted substituent is selected from deuterium, halogen, hydroxy, mercapto, cyano, nitro, acetamido, azido, sulfonyl, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, C_1-8Alkyl, trifluoromethyl, C_2-8Alkenyl radical, C_2-8Alkynyl, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, C_1-8Alkoxy radical, C_1-8Alkoxycarbonyl, C_1-8Alkylcarbonyl group, C_1-8Alkylcarbonyloxy, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Aryl sulfurA group, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, amino, mono C_1-8Alkylamino or di-C_1-8An alkylamino group;

m is 0, 1, 2, 3 or 4;

n is 0, 1, 2, 3, 4 or 5;

r is 0, 1 or 2.

In a preferred embodiment of the present invention, a compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R is₁Selected from hydrogen atoms, halogens, C_1-6Alkyl or C_1-6Haloalkyl, preferably hydrogen atom, halogen.

In a preferred embodiment of the present invention, one of said compounds of formula (I), its stereoisomers or its pharmaceutically acceptable salts thereof is characterized by R, R₃、R₄Each independently selected from hydrogen, halogen or C_1-6An alkyl group; preferably a hydrogen atom.

In a preferred embodiment of the present invention, a compound of said formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R is₂Selected from hydrogen, halogen, cyano, C_1-6Alkoxy, 3-6 membered heterocyclyl or 5-8 membered heteroaryl; wherein said C_1-6Alkoxy, 3-6 membered heterocyclyl and 5-8 membered heteroaryl optionally further substituted by C_1-6Alkyl, - (CH)₂)_xR₅、-(CH₂)_xOR₅、-(CH₂)_xCOR₅、-(CH₂)_xOCOR₅、C_3-8Cycloalkyl and 3-6 membered heterocyclyl, wherein R is₅Selected from hydrogen atom, hydroxyl, acyl, C_1-6Alkyl radical, C_3-8Cycloalkyl, 3-6 membered heterocyclyl and 5-8 membered aryl, and x is an integer from 0 to 6.

In a preferred embodiment of the present invention, one of said compounds of formula (I), its stereoisomers, or its pharmaceutically acceptable salts, is selected from the group consisting of compounds of formula (II):

wherein:

R、R₁、R₂and n is as defined for formula (I);

in a preferred embodiment of the present invention, a compound represented by the formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is characterized by being a compound represented by the formula (III):

wherein:

R_aselected from hydrogen atoms, -C_1-6Alkyl, -C_3-8Cycloalkyl or 3-8 heterocyclyl; wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by-C_1-6Alkyl, -C_3-8Cycloalkyl, -ORc, -CORc, -COORc, -OCORc, 3-8 heterocyclyl, 5-8 heteroaryl, and 5-8 aryl;

R_bselected from hydrogen atoms, halogens or-C_1-6An alkyl group;

R_cselected from hydrogen atoms, -C_1-6Alkyl or 3-8 heterocyclyl;

R、R₁～R₄m and n are as defined in formula (I); and is

x is an integer of 0, 1 or 2.

In a preferred embodiment of the present invention, the compound of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is characterized by being a compound of formula (IV):

wherein:

R₁、R₂、R_a、R_b、R_cx and n are as defined in formula (III)And (5) defining.

As a most preferred embodiment, the compound of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds:

in another aspect, the present invention provides a process for the preparation of a compound of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as hereinbefore described, comprising the steps of:

in another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound as described above, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

Another aspect of the present invention provides the use of a compound as described above, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, for the manufacture of a medicament for the treatment of a disease characterized by the pathology of a tryptophan metabolism disorder mediated by IDO/TDO; the disease which is a pathological feature of the IDO/TDO mediated tryptophan metabolism disorder is preferably selected from cancer or a tumor, a viral infection, depression, a neurodegenerative disorder, trauma, age-related cataract, organ transplant rejection or an autoimmune disease.

Detailed Description

Detailed description: unless stated to the contrary, the following terms used in the specification and claims have the following meanings.

“C_1-8Alkyl "refers to straight and branched alkyl groups comprising 1 to 8 carbon atoms, alkyl refers to a saturated aliphatic hydrocarbon group, C_0-8Means containing no carbon atoms or C_1-8Alkyl, preferably straight-chain alkyl and branched-chain alkyl of 1 to 6 carbon atoms, preferably straight-chain alkyl and branched-chain alkyl of 1 to 3 carbon atoms, for example methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylpentyl, 1, 2-dimethylpropyl, 2-ethylpropyl, 2-dimethylpropyl, 3-dimethylbutyl, 2-ethylpropyl, 2, 2, 3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2-dimethylpentyl, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2-dimethylhexyl, 3-dimethylhexyl, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl or various branched isomers thereof, and the like.

Alkyl groups may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment, preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkene(s)Base, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio;

"cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, "C_3-8Cycloalkyl "refers to a cycloalkyl group comprising 3 to 8 carbon atoms," C_5-10The "cycloalkyl group" means a cycloalkyl group containing 5 to 10 carbon atoms, preferably 5 to 6 carbon atoms_5-6Cycloalkyl radicals ", for example:

non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like.

Polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups. "spirocycloalkyl" refers to polycyclic groups that share a single carbon atom (called a spiro atom) between single rings, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Spirocycloalkyl groups are classified according to the number of spiro atoms shared between rings into mono-, di-or multi-spirocycloalkyl groups, non-limiting examples of which include:

"fused cyclic alkyl" refers to an all-carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. And may be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused ring alkyl groups depending on the number of constituent rings, non-limiting examples of fused ring alkyl groups including:

"bridged cycloalkyl" refers to an all-carbon polycyclic group in which any two rings share two carbon atoms not directly connected, and these may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Depending on the number of constituent rings, may be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl groups, non-limiting examples of which include:

the cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, where the ring to which the parent structure is attached is cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, and the like.

Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio;

"Heterocyclyl" means a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent in which one or more ring atoms are selected from nitrogen, oxygen, or S (O)_r(wherein r is an integer 0, 1, 2) but does not include the ring moiety of-O-O-, -O-S-or-S-S-, the remaining ring atoms being carbon. "5-10 membered heterocyclic group" means a cyclic group containing 5 to 10 ring atoms, "3-8 membered heterocyclic group" means a cyclic group containing 3 to 8 ring atoms, preferably "5-6 membered heterocyclic group" means a cyclic group containing 5 to 6 ring atoms.

Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, propylene oxide, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, and the like, with propylene oxide, piperazinyl, and morpholinyl being preferred.

Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups. "Spiroheterocyclyl" refers to polycyclic heterocyclic groups in which one atom (referred to as a spiro atom) is shared between monocyclic rings, and in which one or more ring atoms are selected from nitrogen, oxygen, or S (O)_r(wherein r is an integer of 0, 1, 2) and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Spirocycloalkyl groups are classified into a single spiroheterocyclyl group, a double spiroheterocyclyl group, or a multiple spiroheterocyclyl group according to the number of spiro atoms shared between rings. Non-limiting examples of spirocycloalkyl groups include:

"fused heterocyclyl" refers to polycyclic heterocyclic groups in which each ring in the system shares an adjacent pair of atoms with other rings in the system, and one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system, wherein one or more ring atoms are selected from nitrogen, oxygen, or S (O)_r(wherein r is an integer of 0, 1, 2) and the remaining ring atoms are carbon. And may be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocycloalkyl depending on the number of rings comprising, non-limiting examples of fused heterocyclic groups include:

"bridged heterocyclyl" means polycyclic heterocyclic groups in which any two rings share two atoms not directly attached, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system in which one or more ring atoms are selected from nitrogen, oxygen, or S (O)_r(wherein r is an integer of 0, 1, 2) and the remaining ring atoms are carbon. According to the number of constituent ringsMesh may be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups, non-limiting examples of which include:

the heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring to which the parent structure is attached is heterocyclyl, non-limiting examples of which include:

the heterocyclic group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio;

"aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group, a polycyclic (i.e., rings which carry adjacent pairs of carbon atoms) group having a conjugated pi-electron system, and a "C" group_5-10Aryl "means an all-carbon aryl group having 5 to 10 carbons, and" 5-to 10-membered aryl "means an all-carbon aryl group having 5 to 10 carbons, such as phenyl and naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

aryl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio;

"heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms including nitrogen, oxygen, and S (O)_r(wherein r is an integer 0, 1, 2), 5-6 membered heteroaryl means a heteroaromatic system containing 5-7 ring atoms, 5-10 membered heteroaryl means a heteroaromatic system containing 5-10 ring atoms, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl and the like, preferably pyrazolyl. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring joined together with the parent structure is a heteroaryl ring, non-limiting examples of which include:

heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio;

"alkenyl" means a radical consisting of at least two carbon atomsAlkyl groups as defined above consisting of at least one carbon-carbon double bond, C_2-8Alkenyl means a straight or branched chain alkenyl group containing 2 to 8 carbons. Such as ethenyl, 1-propenyl, 2-propenyl, 1-, 2-or 3-butenyl, and the like.

The alkenyl group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, or 5-10 membered heteroarylthio;

"alkynyl" means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, C_2-8Alkynyl refers to straight or branched chain alkynyl groups containing 2-8 carbons. For example, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-or 3-butynyl and the like.

Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, or 5-10 membered heteroarylthio;

"alkoxy" refers to-O- (alkyl) wherein alkyl is as defined above. C_1-8Alkoxy means an alkyloxy group having 1 to 8 carbons, and non-limiting examples include methoxy, ethoxy, propoxy, butoxy, and the like.

Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents, preferablyIs one or more groups independently selected from halogen, hydroxyl, sulfhydryl, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, or 5-10 membered heteroarylthio;

"halogen-substituted C_1-8Alkyl "refers to a 1-8 carbon alkyl group optionally substituted with fluorine, chlorine, bromine, iodine atoms for hydrogen on the alkyl, such as difluoromethyl, dichloromethyl, dibromomethyl, trifluoromethyl, trichloromethyl, tribromomethyl, and the like.

"halogen-substituted C_1-8Alkoxy "refers to a 1-8 carbon alkoxy group wherein the hydrogen on the alkyl group is optionally substituted with fluorine, chlorine, bromine, or iodine atoms. For example, difluoromethoxy, dichloromethoxy, dibromomethoxy, trifluoromethoxy, trichloromethoxy, tribromomethoxy and the like.

"halogen" refers to fluorine, chlorine, bromine or iodine.

"DMSO" refers to dimethyl sulfoxide.

"LDA" refers to lithium diisopropylamide.

"DIAD" refers to diisopropyl azodicarboxylate.

"DMF" refers to N, N-dimethylformamide.

"DPPA" refers to diphenylphosphoryl azide.

“Pd(dppf)Cl₂"means [1, 1' -bis (diphenylphosphino) ferrocene]Palladium dichloride.

"THF" refers to tetrahydrofuran.

"optionally substituted" means that one or more hydrogen atoms in a group are independently replaced by a corresponding number of deuterium, halogen, hydroxy, mercapto, cyano, nitro, azido, C_1-8Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, halo-substituted C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heteroheteroCyclylthio group, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, or 5-10 membered heteroarylthio; preferably deuterium, halogen, hydroxy, mercapto, cyano, nitro, acetylamino, azido, sulfonyl, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, C_1-8Alkyl, trifluoromethyl, C_2-8Alkenyl radical, C_2-8Alkynyl, C_3-8Cycloalkyl, 3-8 membered heterocyclyl, C_1-8Alkoxy radical, C_1-8Alkoxycarbonyl, C_1-8Alkylcarbonyl group, C_1-8Alkylcarbonyloxy, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C_5-10Aryl radical, C_5-10Aryloxy radical, C_5-10Arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, amino, mono C_1-8Alkylamino or di-C_1-8Substituted by a substituent of alkylamino.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "a heterocyclic group optionally substituted with an alkyl" means that an alkyl may, but need not, be present, and the description includes the case where the heterocyclic group is substituted with an alkyl and the heterocyclic group is not substituted with an alkyl.

"substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable in combination with carbon atoms having unsaturated bonds (e.g., olefinic).

"pharmaceutical composition" means a mixture containing one or more compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof in admixture with other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient and exert biological activity.

The present invention will be described more fully with reference to the following examples, but the present invention is not limited thereto, and the present invention is not limited to the examples.

The structure of the compounds of the invention is determined by Nuclear Magnetic Resonance (NMR) or/and liquid mass chromatography (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR was measured using a Bruker AVANCE-400 NMR spectrometer using deuterated methanol (CD)₃OD) and deuterated chloroform (CDCl)₃) Internal standard is Tetramethylsilane (TMS).

LC-MS was measured using an Agilent 1200 Infinity Series Mass spectrometer. HPLC was carried out using an Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18150X 4.6mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18150X 4.6mm column).

The thin layer chromatography silica gel plate adopts a tobacco yellow sea HSGF254 or Qingdao GF254 silica gel plate, the specification adopted by TLC is 0.15 mm-0.20 mm, and the specification adopted by the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm. The column chromatography generally uses 200-300 mesh silica gel of the Tibet Huanghai silica gel as a carrier.

The starting materials in the examples of the present invention are known and commercially available, or may be synthesized using or according to methods known in the art.

All reactions of the present invention are carried out under a dry nitrogen or argon atmosphere with continuous magnetic stirring, and the solvent is a dry solvent, unless otherwise specified.

An argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon of argon or nitrogen with a volume of about 1L. The hydrogen atmosphere refers to a reaction flask connected with a hydrogen balloon with a volume of about 1L.

The solutions in the examples are aqueous solutions unless otherwise specified. The reaction temperature was room temperature. The room temperature is the most suitable reaction temperature and is 20-30 ℃.

The progress of the reaction in the examples was monitored by Thin Layer Chromatography (TLC) or liquid chromatography-mass spectrometry (LC-MS) using the following developer systems: the volume ratio of dichloromethane and methanol system, n-hexane and ethyl acetate system, petroleum ether and ethyl acetate system, acetone and solvent can be regulated according to different polarities of the compounds. The system of eluent for column chromatography comprises: a: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: dichloromethane and ethyl acetate system, D: ethyl acetate and methanol, the volume ratio of the solvent is adjusted according to the different polarities of the compounds, and a small amount of ammonia water, acetic acid and the like can be added for adjustment.

Example 1:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-cyanophenyl) urea

The first step is as follows: preparation of 1- (2-bromophenylmethyl) -1H-imidazole

O-bromotoluene (20g, 117mmol), bromosuccinimide (22.0g, 123mmol), azobisisobutyronitrile (1g) were dissolved in dry carbon tetrachloride (200mL) and heated under reflux for 16 hours under nitrogen. The reaction was cooled to room temperature and filtered, imidazole (15.9g, 234mmol) and potassium carbonate (33g, 234mmol) were added to the filtrate, and the mixture was heated under reflux for 5 hours under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (100mL) and washed with water (2X 50 mL). The organic phase was separated, dried over anhydrous sodium sulfate, filtered, concentrated and the residue was purified by flash silica gel column to give 1- (2-bromophenylmethyl) -1H-imidazole (20 g).

LC-MS：236.9，239.1，t_R＝1.541min.

The second step is that: 5H-imidazo [5, 1-a ]]Preparation of isoindoles

1- (2-bromo-benzyl) -1H-imidazole (20g, 84.0mmol), palladium acetate (500mg), triphenylphosphine (500mg), potassium carbonate (23g, 167mmol) were dissolved in DMSO (200mL) and heated to 140 ℃ under nitrogen with stirring for 2H. LC-MS showed the reaction was complete, the reaction was concentrated to dryness, and the residue was dissolved in ethyl acetate (200mL) and washed successively with water (2X 100mL), saturated brine (50 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 5H-imidazo [5, 1-a ] isoindole (10.0 g).

LC-MS：157.1，t_R＝1.466min.

The third step: (5H-imidazo [5, 1-a ]]Preparation of isoindol-5-yl) methanol

5H-imidazo [5, 1-a ] isoindole (668mg, 4.277mmol), paraformaldehyde (193mg, 6.427mmol) were stirred in tetrahydrofuran (5mL), cooled to-78 ℃ under nitrogen, and LDA (3.2mL, 2M in THF) was added dropwise. Slowly warm to room temperature and stir overnight. Adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain (5H-imidazo [5, 1-a ] isoindol-5-yl) methanol (196 mg).

LC-MS：187.1，t_R＝1.396min.

The fourth step: 2- ((5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) methyl) isoindoline-1, 3-dione

(5H-imidazo [5, 1-a ] isoindol-5-yl) methanol (180mg, 0.967mmol), phthalimide (294mg, 2.0mmol), and triphenylphosphine (525mg, 2.0mmol) were dissolved in toluene (5mL), stirred under nitrogen, and a solution of DIAD (404mg, 2.0mmol) in toluene (5mL) was added dropwise at room temperature and stirred overnight. Filtering, extracting the filtrate with water and ethyl acetate, drying, and purifying by column chromatography to obtain 2- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) isoindoline-1, 3-dione (93 mg).

LC-MS：316.0，t_R＝1.866min.

The fifth step: (5H-imidazo [5, 1-a ]]Preparation of isoindol-5-yl) methylamines

2- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) isoindoline-1, 3-dione (93mg, 0.295mmol), hydrazine hydrate (85%, 1mL) were refluxed in ethanol (2mL) for 4 hours, cooled, concentrated, and the residue was purified by column chromatography to give (5H-imidazo [5, 1-a ] isoindol-5-yl) methylamine (20 mg).

LC-MS：186.1，t_R＝0.618min.

And a sixth step: 1- ((5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) methyl) -3- (4-cyanophenyl) urea

(5H-imidazo [5, 1-a ] isoindol-5-yl) methylamine (20mg, 0.108mmol), 4-cyanobenzene isocyanate (20mg, 0.139mmol), triethylamine (22mg, 0.217mmol) were stirred in DMF (2mL) at room temperature overnight, water was added, dichloromethane was added for extraction, dried, and purified by column chromatography to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-cyanophenyl) urea (5 mg).

LC-MS：330.1，t_R＝1.896min；

¹H NMR(400MHz，d₆-DMSO)δ9.25(s，1H)，8.24(s，1H)，7.72-7.57(m，4H)，7.56-7.41(m，3H)，7.39-7.27(m，2H)，6.64(t，J＝5.9Hz，1H)，5.61-5.46(m，1H)，

4.07-3.94(m，1H)，3.66-3.55(m，1H).

Example 2:

1- (4-cyanophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

The first step is as follows: preparation of 1- (2-fluoro-6-bromophenylmethyl) -1H-imidazole

1-bromo-3-fluoro-2-toluene (8.0g, 42.3mmol), bromosuccinimide (9.0g, 50.8mmol), azobisisobutyronitrile (300mg) were dissolved in dry carbon tetrachloride (100mL) and heated under reflux for 16 hours under nitrogen. The reaction was cooled to room temperature and filtered, imidazole (8.7g, 126.9mmol) and potassium carbonate (17.5g, 126.9mmol) were added to the filtrate, and the mixture was heated under reflux for 5 hours under nitrogen. LC-MS showed complete reaction, the reaction was filtered, the filtrate was concentrated, and the residue was purified by flash column chromatography on silica gel to give 1- (2-bromo-6-fluorobenzyl) -1H-imidazole (6.0 g).

The second step is that: 6-fluoro-5H-imidazo [5, 1-a]Preparation of isoindoles

1- (2-bromo-6-fluorobenzyl) -1H-imidazole (6.0g, 23.5mmol), palladium acetate (200mg), triphenylphosphine (300mg), potassium carbonate (6.5g, 47.0mmol) were dissolved in DMSO (50mL) and the reaction was heated to 140 ℃ under nitrogen and stirred for 2H. LC-MS showed the reaction was complete, the reaction was concentrated to dryness, and the residue was dissolved in ethyl acetate (200mL) and washed successively with water (2X 100mL), saturated brine (50 mL). The organic phase was dried, filtered, concentrated and the residue was purified by flash column chromatography on silica gel to give 6-fluoro-5H-imidazo [5, 1-a ] isoindole (3.0 g).

The third step: 2- (6-fluoro-5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) acetic acid tert-butyl ester

6-fluoro-5H-imidazo [5, 1-a ] isoindole (200mg, 1.148mmol) is dissolved in dry tetrahydrofuran (2mL), cooled to-78 ℃ under nitrogen and LDA (0.86mL, 2M in THF) is added dropwise. Slowly raising the temperature to room temperature, stirring for 3 hours, adding water, extracting by dichloromethane, drying, and purifying by column chromatography to obtain tert-butyl 2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetate (245 mg).

LC-MS：289.1，t_R＝1.989min.

The fourth step: 2- (6-fluoro-5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) acetic acid

Tert-butyl 2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetate (245mg, 0.850mmol) was dissolved in ethyl acetate (2mL), and a hydrogen chloride/dioxane solution (2mL) was added thereto, followed by stirring overnight at room temperature and purification by column chromatography to give 2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (93 mg).

LC-MS：233.1，t_R＝1.532min.

The fifth step: 1- (4-cyanophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Process for preparing isoindol-5-yl) methyl) urea Preparation of

2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (30mg, 0.129mmol), 4-cyanoaniline (17mg, 0.144mmol), DPPA (39mg, 0.142mmol), triethylamine (20mg, 0.198mmol) were refluxed in toluene (4mL) under nitrogen for 4 hours, cooled to room temperature, extracted with water, dichloromethane, dried, and purified by column chromatography to give 1- (4-cyanophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (11 mg).

LC-MS：348.1，t_R＝1.853min；

¹H NMR(400MHz，d₆-DMSO)δ9.57(s，1H)，9.17(s，1H)，7.83(s，1H)，7.62(ddd，J＝15.6，12.4，7.7Hz，4H)，7.49(d，J＝8.7Hz，2H)，7.33(t，J＝9.0Hz，1H)，7.16(s，1H)，5.99(s，1H)，4.21(d，J＝15.0Hz，1H)，3.69(d，J＝8.2Hz，1H).

Example 3:

1- (4-bromophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (200mg, 0.86mmol), Diphenylphosphorylazide (DPPA) (475mg, 1.72mmol), triethylamine (180mg, 1.72mmol), and p-bromoaniline (300mg, 1.72mmol) were dissolved in anhydrous toluene (20mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (4-bromophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (6.0 mg).

LC-MS：401.0，t_R＝2.118min；

¹H NMR(400MHz，MeOD)δ7.96(s，1H)，7.56-7.43(m，2H)，7.42-7.31(m，2H)，7.27-7.18(m，3H)，7.15-7.03(m，1H)，5.77-5.61(m，1H)，4.22(dd，J＝14.4，3.9Hz，1H)，3.68(dd，J＝14.4，5.9Hz，1H).

Example 4:

1- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyrazol-4-yl) Phenyl) urea

The first step is as follows: preparation of 4- (1-methyl-1H-pyrazol-4-yl) aniline

P-bromoaniline (2.0g, 11.5mmol), 1-methylpyrazole-4-boronic acid pinacol ester (2.4g, 11.5mmol), Pd (dppf) Cl₂(100mg), potassium phosphate (4.8g, 23.0mmol) was dissolved in 1, 4-dioxane (20mL) and 20mL water, heated to 100 ℃ under nitrogen and stirred for 3 hours. LC-MS showed the reaction was complete. The reaction was concentrated and the residue was dissolved in dichloromethane (50mL) and washed with water (2X 30 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 4- (1-methyl-1H-pyrazol-4-yl) aniline (1.5 g).

The second step is that: 1- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyri dine Preparation of oxazol-4-yl) phenyl) urea

2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (200mg, 0.86mmol), Diphenylphosphorylazide (DPPA) (475mg, 1.72mmol), triethylamine (180mg, 1.72mmol), and 4- (1-methyl-1H-pyrazol-4-yl) aniline (300mg, 1.72mmol) were dissolved in anhydrous toluene (20mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (20.8 mg).

LC-MS：403.15，t_R＝1.831min；

¹H NMR(400MHz，MeOD)δ7.97(s，1H)，7.86(s，1H)，7.74(s，1H)，7.53-7.45(m，2H)，7.44-7.38(m，2H)，7.30-7.25(m，2H)，7.22(s，1H)，7.09(ddd，J＝9.5，6.3，2.9Hz，1H)，5.69(dd，J＝5.8，4.0Hz，1H)，4.22(dd，J＝14.4，3.9Hz，1H)，3.91(s，3H)，3.66.

Example 5：

1- (4-cyanophenyl) -3- ((6- (trifluoromethyl) -5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

The first step is as follows: preparation of 1- (2-bromo-6- (trifluoromethyl) benzyl) -1H-imidazole

1-bromo-3- (trifluoromethyl) -2-toluene (5.0g, 20.9mmol), bromosuccinimide (3.72g, 20.9mmol), azobisisobutyronitrile (300mg) were dissolved in dry carbon tetrachloride (100mL) and heated under reflux for 16 hours under nitrogen. The reaction was cooled to room temperature and filtered, imidazole (2.85g, 42mmol) and potassium carbonate (4.34g, 31.37mmol) were added to the filtrate, and the mixture was heated under reflux for 5 hours under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (50mL) and washed with water (2X 40 mL). The organic phase was dried, filtered and concentrated, and the residue was purified by flash column chromatography on silica gel to give 1- (2-bromo-6- (trifluoromethyl) benzyl) -1H-imidazole (5.0 g).

The second step is that: 6- (trifluoromethyl) -5H-imidazo [5, 1-a]Preparation of isoindoles

1- (2-bromo-6- (trifluoromethyl) benzyl) -1H-imidazole (5.0g, 16.4mmol), palladium acetate (184mg), triphenylphosphine (430mg), potassium carbonate (4.4g, 32.8mmol) were dissolved in DMSO (50mL) and heated to 140 ℃ under nitrogen with stirring for 2H. LC-MS showed the reaction was complete, the reaction was concentrated to dryness, and the residue was dissolved in ethyl acetate (200mL) and washed successively with water (2X 100mL), saturated brine (50 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column on silica gel to give 6- (trifluoromethyl) -5H-imidazo [5, 1-a ] isoindole (2.9 g).

The third step: 2- (6- (trifluoromethyl) -5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) acetic acid tert-butyl ester

Reacting 6- (trifluoromethyl) -5H-imidazo [5, 1-a ]]Isoindole (2.9g, 12.9mmol) was dissolved in anhydrous THF (100mL) and LDA (9.7mL, 19.4mmol, 2M in THF) was slowly added dropwise at-30 ℃. After completion of the dropwise addition, the mixture was stirred at-30 ℃ for 15 minutes, and tert-butyl bromoacetate (3.8g, 19.4mmol) was slowly added dropwise thereto. Stirring at-30 deg.C to 0 deg.C for 1 hr, LC-MS indicating reaction is complete, and saturated NH₄Cl (100 mL). The organic phase is dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 2- (6- (trifluoromethyl) -5H-imidazo [5, 1-a)]Isoindol-5-yl) acetic acid tert-butyl ester (2 g).

The fourth step: 2- (6- (trifluoromethyl) -5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) acetic acid

Tert-butyl 2- (6- (trifluoromethyl) -5H-imidazo [5, 1-a ] isoindol-5-yl) acetate (230mg, 0.68mmol) was dissolved in trifluoroacetic acid (2mL), dichloromethane (2mL), stirred at room temperature for 1 hour, and the reaction solution was concentrated to give crude 2- (6- (trifluoromethyl) -5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (200 mg).

The fifth step: 1- (4-cyanophenyl-3- ((6- (trifluoromethyl) -5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl Yl) preparation of urea

2- (6- (trifluoromethyl) -5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (200mg, 0.71mmol), Diphenylphosphorylazide (DPPA) (487mg, 1.77mmol), triethylamine (0.4mL, 2.83mmol), 4-aminophenylacetonitrile (0.21mg, 1.77mmol) were dissolved in anhydrous toluene (20mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (4-cyanophenyl-3- ((6- (trifluoromethyl) -5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (6.1 mg).

LC-MS：398.0，t_R＝2.044Min；

¹H NMR(400MHz，MeOD)δ7.99(s，1H)，7.93(d，J＝5.5Hz，1H)，7.73-7.64(m，2H)，7.57(d，J＝8.8Hz，2H)，7.46(d，J＝8.8Hz，2H)，7.27(s，1H)，5.88-5.82(m，1H)，4.35(dd，J＝14.6，2.9Hz，2H)，3.57(dd，J＝14.6，5.8Hz，2H)；

¹⁹F NMR(400MHz，MeOD)δ-60.9.

Example 6：

1- ((6-chloro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-cyanophenyl) urea

2- (6-chloro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (420mg, 1.69mmol) was dissolved in 15mL of toluene, and DPPA (511mg, 1.859mmol), triethylamine (256mg, 2.533mmol) and p-cyanoaniline (219mg, 1.859mmol) were added. Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction was concentrated, and the residue was purified by flash column chromatography on silica gel (dichloromethane: methanol ═ 10: 1) to give 1- ((6-chloro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-cyanophenyl) urea (58 mg).

LC-MS：364.0，t_R＝1.926min；

¹H NMR(400MHz，d₆-DMSO)δ9.02(s，1H)，7.93(s，1H)，7.63(dd，J＝15.8，8.1Hz，3H)，7.53-7.42(m，3H)，7.34(dd，J＝7.7，4.2Hz，1H)，7.21(s，1H)，6.44(t，J＝6.0Hz，1H)，5.61(dd，J＝6.1，3.3Hz，1H)，4.28(ddd，J＝14.1，5.7，3.4Hz，1H)，3.65-3.51(m，1H).

Example 7：

1- (3-fluoro-4-methoxyphenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (80mg, 0.35mmol), Diphenylphosphorylazide (DPPA) (190mg, 0.69mmol), triethylamine (100mg, 0.9mmol), and 3-fluoro-4-methoxyaniline (100mg, 0.69mmol) were dissolved in anhydrous toluene (10mL), and the mixture was refluxed under nitrogen for 4 hours. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (3-fluoro-4-methoxyphenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (16.0 mg).

LC-MS：371.0，t_R＝1.970min；

¹H NMR(400MHz，MeOD)δ7.96(s，1H)，7.54-7.44(m，2H)，7.20(dd，J＝13.4，2.4Hz，2H)，7.09(ddd，J＝9.5，6.5，2.6Hz，1H)，6.96(t，J＝9.0Hz，1H)，6.88(ddd，J＝8.8，2.4，1.3Hz，1H)，5.69(dd，J＝5.7，4.1Hz，1H)，4.20(dd，J＝14.4，3.9Hz，1H)，3.82(s，3H)，3.66(dd，J＝14.4，6.0Hz，1H).

Example 8:

1- (3-bromophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (80mg, 0.35mmol), Diphenylphosphorylazide (DPPA) (190mg, 0.69mmol), triethylamine (100mg, 0.9mmol), and 3-bromoaniline (120mg, 0.69mmol) were dissolved in anhydrous toluene (10mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (3-bromophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (2.1 mg).

LC-MS：401.0，t_R＝2.11min；

¹H NMR(400MHz，MeOD)δ7.97(s，1H)，7.63(s，1H)，7.48(s，2H)，7.15(dd，J＝26.1，17.6Hz，4H)，5.70(s，1H)，4.22(d，J＝14.0Hz，1H)，3.78-3.54(m，1H).

Example 9:

1- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-fluorophenyl) urea

2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (80mg, 0.35mmol), Diphenylphosphorylazide (DPPA) (190mg, 0.69mmol), triethylamine (100mg, 0.9mmol), and 4-fluoroaniline (80mg, 0.69mmol) were dissolved in anhydrous toluene (10mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-fluorophenyl) urea (11.2 mg).

LC-MS：341.0，2.116min；

¹H NMR(400MHz MeOD)δ7.98(s，1H)，7.92-7.81(m，1H)，7.53-7.41(m，2H)，7.22(s，1H)，7.08(ddd，J＝17.2，9.7，5.1Hz，3H)，7.00-6.93(m，1H)，5.79-5.62(m，1H)，4.24(dd，J＝14.4，3.8Hz，1H)，3.73(dd，J＝14.5，5.6Hz，1H).

Example 10:

1- (4- (1-methyl-1H-pyrazol-4-yl) phenyl) -3- ((6- (trifluoromethyl) -5H-imidazo [5, 1-a)]Isoindole derivatives Indol-5-yl) methyl) urea

The preparation method is the same as example 5.

LC-MS：453.1，t_R＝1.983min；

¹H NMR(400MHz，MeOD)δ7.86(s，1H)，7.83-7.77(m，1H)，7.74(s，1H)，7.61(s，1H)，7.59-7.49(m，2H)，7.33-7.25(m，2H)，7.18-7.09(m，3H)，5.71(s，1H)，4.21(dd，J＝14.5，3.4Hz，1H)，3.79(s，3H)，3.39(dd，J＝14.5，6.1Hz，1H)；

¹⁹F NMR(400MHz，MeOD)δ-60.9.

Example 11:

1- ((6-chloro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyrazol-4-yl) Phenyl) urea

2- (6-chloro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (245mg, 0.984mmol) is dissolved in 15mL of toluene, followed by the addition of DPPA (298mg, 1.083mmol), triethylamine (149mg, 1.478mmol), and 4- (1-methyl-1H-pyrazol-4-yl) aniline (188mg, 1.083 mmol). Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction mixture was concentrated, and the residue was purified by flash column chromatography on silica gel (dichloromethane: methanol ═ 10: 1) to give 1- ((6-chloro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (50 mg).

LC-MS：419.0，t_R＝1.920min；

¹H NMR(400MHz，d₆-DMSO)δ8.48(s，1H)，7.96(d，J＝23.3Hz，2H)，7.74(s，1H)，7.61(d，J＝7.4Hz，1H)，7.45(t，J＝7.8Hz，1H)，7.45-7.23(m，5H)，7.22(s，1H)，6.20(t，J＝5.9Hz，1H)，5.60(dd，J＝6.1，3.2Hz，1H)，4.29(ddd，J＝14.1，5.7，3.5Hz，1H)，3.83(s，3H)，3.58-3.47(m，1H).

Example 12:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-2-fluorophenyl) urea

The first step is as follows: 2- (5H-imidazo [5, 1-a ]]Isoindol-5-yl) ethanesPreparation of tert-butyl ester

Reacting 5H-imidazo [5, 1-a ]]Isoindole (1.0g, 6.41mmol) was dissolved in anhydrous THF (30mL) and LDA (4.8mL, 9.61mmol, 2M in THF) was slowly added dropwise at-30 ℃. After completion of the dropwise addition, the reaction was stirred at-30 ℃ for 20 minutes, and tert-butyl bromoacetate (1.3g, 7.05mmol) was slowly added dropwise. The reaction is stirred for 2 hours at the temperature of between 30 ℃ below zero and 0 ℃, LC-MS shows that the reaction is complete, and saturated NH is used for the reaction₄Cl (10 mL). The organic phase is dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 2- (5H-imidazo [5, 1-a)]Isoindol-5-yl) acetic acid tert-butyl ester (250 mg).

The second step is that: 2- (5H-imidazo [5, 1-a ]]Preparation of isoindol-5-yl) acetic acid

Tert-butyl 2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetate (200mg, 0.71mmol) was dissolved in trifluoroacetic acid (2mL), dichloromethane (2mL), stirred at room temperature for 1 hour, and the reaction mixture was concentrated to give crude 2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (150 mg).

The third step: 1- ((5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) methyl) -3- (4-bromo-2-fluorophenyl) urea Prepare for

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (200mg, 0.934mmol), 4-bromo-2-fluoroaniline (213mg, 1.121mmol), DPPA (308mg, 1.119mmol) and triethylamine (142mg, 1.403mmol) were dissolved in toluene (20mL) under nitrogen and refluxed for 4 hours, cooled to room temperature, extracted with water, dichloromethane, dried, and purified by column chromatography to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-bromo-2-fluorophenyl) urea (97 mg).

LC-MS：401.0，403.0，t_R＝2.052min；

¹H NMR(400MHz，MeOD)δ8.02-7.84(m，2H)，7.62(dd，J＝23.6，7.6Hz，2H)，7.45(t，J＝7.4Hz，1H)，7.40-7.10(m，4H)，5.49(t，J＝4.8Hz，1H)，4.12(dd，J＝14.4，4.1Hz，1H)，3.66(dd，J＝14.4，5.8Hz，1H).

Example 13:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2-fluoro-4- (1-methyl-1H-pyrazol-4-yl) Phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-2-fluorophenyl) urea (45mg, 0.112mmol), 1-methyl-1H-pyrazole-4-boronic acid pinacol ester (47mg, 0.226mmol), Pd (dppf) Cl₂(40mg, 0.055mmol) and potassium carbonate (31mg, 0.224mmol) were dissolved in 1, 4-dioxane/water (5mL/1mL) and stirred, and the temperature was raised to 110 ℃ under nitrogen protection for 1 hour by microwave reaction. Cooling to room temperature, adding 20ml water, extracting with dichloromethane, drying, purifying by column chromatography to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2-fluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (1.1 mg).

LC-MS：403.1，t_R＝1.885min；

¹H NMR(400MHz，MeOD)δ7.93(dq，J＝17.3，8.8Hz，3H)，7.78(s，1H)，7.63(dd，J＝22.7，7.5Hz，2H)，7.51-7.24(m，4H)，7.19(s，1H)，5.55-5.46(m，1H)，4.12(dd，J＝14.3，4.1Hz，1H)，3.92(s，3H)，3.64(dd，J＝14.3，6.1Hz，1H).

Example 14:

1- (4-cyanophenyl) -3- ((7-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) Urea

The first step is as follows: preparation of 1- (2-bromo-5-fluoro) benzyl) -1H-imidazole

1-bromo-4-fluoro-2-toluene (5.0g, 26.5mmol), bromosuccinimide (5.65g, 31.7mmol), azobisisobutyronitrile (400mg) were dissolved in dry carbon tetrachloride (100mL) and heated under reflux for 16 hours under nitrogen. The reaction was cooled to room temperature and filtered, imidazole (3.65g, 52.3mmol) and potassium carbonate (5.42g, 39.2mmol) were added to the filtrate, and the mixture was heated under reflux for 5 hours under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (50mL) and washed with water (2X 40 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 1- (2-bromo-5-fluoro) benzyl) -1H-imidazole (5.0 g).

The second step is that: 7-fluoro-5H-imidazo [5, 1-a]Preparation of isoindoles

1- (2-bromo-5-fluoro) benzyl) -1H-imidazole (1.5g, 5.9mmol), palladium acetate (66mg), triphenylphosphine (154mg) and potassium carbonate (1.6g, 11.8mmol) were dissolved in DMSO (30mL) and heated to 140 ℃ under nitrogen with stirring for 2H. LC-MS showed the reaction was complete, the reaction was concentrated to dryness, and the residue was dissolved in ethyl acetate (100mL) and washed successively with water (2X 100mL), saturated brine (50 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 7-fluoro-5H-imidazo [5, 1-a ] isoindole (0.8 g).

The third step: 2- (7-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) ethanesPreparation of tert-butyl ester

Reacting 7-fluoro-5H-imidazo [5, 1-a ]]Isoindole (0.8g, 4.6mmol) was dissolved in anhydrous THF (50mL) and LDA (3.4mL, 6.9mmol, 2M in THF) was slowly added dropwise at-30 ℃. After completion of the dropwise addition, the mixture was stirred at-30 ℃ for 15 minutes, and tert-butyl bromoacetate (1.4g, 6.9mmol) was slowly added dropwise thereto. Stirring at-30 deg.C to 0 deg.C for 1 hr, LC-MS indicating reaction is complete, reaction using saturated NH₄Cl (20 mL). The organic phase is dried, filtered and concentrated. The residue was purified by flash silica gel column to give 2- (7-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) acetic acid tert-butyl ester (0.5 g).

The fourth step: 2- (7-fluoro-5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) acetic acid

Tert-butyl 2- (7-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetate (400mg, 1.4mmol) was dissolved in trifluoroacetic acid (2mL) and dichloromethane (2mL), stirred at room temperature for 1 hour, and the reaction mixture was concentrated to give crude 2- (7-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (300 mg).

The fifth step: 1- (4-cyanophenyl-3- ((7-fluoro-5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) methyl) urea Prepare for

2- (7-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (300mg, 0.71mmol), Diphenylphosphorylazide (DPPA) (487mg, 1.77mmol), triethylamine (0.4mL, 2.83mmol) and 4-aminophenylacetonitrile (0.21mg, 1.77mmol) were dissolved in anhydrous toluene (20mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (4-cyanophenyl-3- ((7-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (0.10 mg).

LC-MS：348.1，t_R＝1.93min；

¹H NMR(400MHz，MeOD)δ7.95(s，1H)，7.64(dd，J＝8.4，4.9Hz，1H)，7.59(d，J＝8.8Hz，2H)，7.52(d，J＝8.8Hz，2H)，7.40(dd，J＝8.8，2.0Hz，1H)，7.20(td，J＝9.0，2.3Hz，1H)，7.14(s，1H)，5.51(t，J＝5.1Hz，1H)，4.10(dd，J＝14.4，4.2Hz，1H)，3.70(dd，J＝14.4，5.8Hz，1H)；

¹⁹F NMR(400MHz，MeOD)δ-116.1.

Example 15:

1- (4-cyanophenyl) -3- ((8-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

The first step is as follows: preparation of 1- (2-bromo-4-fluoro) benzyl) -1H-imidazole

2-bromo-4-fluoro-1-toluene (5.0g, 26.5mmol), bromosuccinimide (5.65g, 31.7mmol), azobisisobutyronitrile (400mg) were dissolved in dry carbon tetrachloride (100mL) and heated under reflux for 16 hours under nitrogen. The reaction was cooled to room temperature and filtered, imidazole (3.65g, 52.3mmol) and potassium carbonate (5.42g, 39.2mmol) were added to the filtrate, and the mixture was heated under reflux for 5 hours under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (50mL) and washed with water (2X 40 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 1- (2-bromo-4-fluoro) benzyl) -1H-imidazole (4.0 g).

The second step is that: 8-fluoro-5H-imidazo [5, 1-a]Preparation of isoindoles

1- (2-bromo-4-fluoro) benzyl) -1H-imidazole (3g, 11.0mmol), palladium acetate (132mg), triphenylphosphine (308mg) and potassium carbonate (3.2g, 23.6mmol) were dissolved in DMSO (50mL) and heated to 140 ℃ under nitrogen with stirring for 2H. LC-MS showed the reaction was complete, the reaction was concentrated to dryness, and the residue was dissolved in ethyl acetate (100mL) and washed successively with water (2X 100mL), saturated brine (50 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel to give 8-fluoro-5H-imidazo [5, 1-a ] isoindole (1.6 g).

The third step: 2- (8-fluoro-5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) acetic acid tert-butyl ester

Reacting 8-fluoro-5H-imidazo [5, 1-a ]]Isoindole (1g, 5.8mmol) was dissolved in anhydrous THF (50mL) and LDA (4.3mL, 8.6mmol, 2M in THF) was slowly added dropwise at-30 ℃. After completion of the dropwise addition, the mixture was stirred at-30 ℃ for 15 minutes, and tert-butyl bromoacetate (1.6g, 8.6mmol) was slowly added dropwise thereto. Stirring at-30 deg.C to 0 deg.C for 1 hr, LC-MS indicating reaction is complete, reaction using saturated NH₄Cl (20 mL). The organic phase is dried, filtered and concentrated. Purifying the residue with flash silica gel column to obtain 2- (8-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) acetic acid tert-butyl ester (0.8 g).

The fourth step: 2- (8-fluoro-5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) acetic acid

Tert-butyl 2- (8-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetate (400mg, 1.4mmol) was dissolved in trifluoroacetic acid (2mL) and dichloromethane (2mL), stirred at room temperature for 1 hour, and the reaction mixture was concentrated to give crude 2- (8-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (300 mg).

The fifth step: 1- (4-cyanophenyl-3- ((8-fluoro-5H-imidazo [5, 1-a)]Preparation of isoindol-5-yl) methyl) urea Prepare for

2- (7-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (300mg, 0.71mmol), Diphenylphosphorylazide (DPPA) (487mg, 1.77mmol), triethylamine (0.4mL, 2.83mmol), 4-aminophenylacetonitrile (0.21mg, 1.77mmol) were dissolved in anhydrous toluene (20mL), and the mixture was refluxed under nitrogen for 4 hours. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (4-cyanophenyl-3- ((8-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (0.10 mg).

LC-MS：348.1，t_R＝1.90min；

¹H NMR(400MHz，MeOD)δ7.98(s，1H)，7.70-7.55(m，3H)，7.55-7.48(m，2H)，7.41(dd，J＝8.7，2.4Hz，1H)，7.22(s，1H)，7.15-7.03(m，1H)，5.49(t，J＝4.8Hz，1H)，4.11(dd，J＝14.4，4.1Hz，1H)，3.66(dd，J＝14.4，5.9Hz，1H)；¹⁹F NMR(400MHz，MeOD)δ-115.0(s，1H).

Example 16:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyrazol-4-yl) phenyl) Urea

1- ((6-chloro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (40mg, 0.095mmol) was dissolved in 8mL of methanol, and then 10mg of Pd-C was added to react under hydrogen at room temperature for 3 days. The LC-MS detection reaction is complete. The reaction solution was filtered, the filtrate was spin-dried to give a crude product, which was subjected to flash column chromatography to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (15mg) as a pale yellow solid.

LC-MS：385.1，t_R＝1.824min；

¹H NMR(400MHz，MeOD)δ8.07(s，1H)，7.72(d，J＝12.9Hz，2H)，7.64(d，J＝7.3Hz，2H)，7.57(d，J＝7.6Hz，1H)，7.50-7.29(m，6H)，7.23(s，1H)，5.57-5.48(m，1H)，4.10(dd，J＝14.4，4.3Hz，1H)，3.93(s，3H)，3.46(dd，J＝14.4，7.1Hz，1H).

Example 17:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-methoxyphenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (150mg, 0.70mmol), Diphenylphosphorylazide (DPPA) (400mg, 1.42mmol), triethylamine (180mg, 1.42mmol) and 3-methoxy-4-bromoaniline (284mg, 1.42mmol) were dissolved in anhydrous toluene (20mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-bromo-3-methoxyphenyl) urea (19.0 mg).

LC-MS：413.0，t_R＝2.052min；

¹H NMR(400MHz，MeOD)δ7.95(s，1H)，7.62(dd，J＝22.4，7.6Hz，2H)，7.45(t，J＝7.5Hz，1H)，7.40-7.31(m，2H)，7.28(d，J＝2.1Hz，1H)，7.18(s，1H)，6.70(dd，J＝8.5，2.2Hz，1H)，5.56-5.43(m，1H)，4.10(dd，J＝14.3，4.2Hz，1H)，3.85(s，3H)，3.60(dd，J＝14.4，6.3Hz，1H).

Example 18:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-methoxy-4- (1-methyl-1H-pyrazole- 4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-methoxyphenyl) urea (30mg, 0.07mmol), 1-methyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (32mg, 0.15mmol), Pd (dppf) Cl₂(5mg), potassium carbonate (40mg, 0.28mmol) was dissolved in dioxane (3mL) and water (1.5mL) and heated under reflux for 2h under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (10mL) and washed with water (2X 5 mL). The organic phase was dried, filtered and concentrated. Purifying the residue with flash silica gel column to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-methoxy-4- (1-methyl-1H-]Pyrazol-4-yl) phenyl) urea (3.4 mg).

LC-MS：415.1，t_R＝2.052min；

¹H NMR(400MHz，MeOD)δ7.95(s，2H)，7.82(s，1H)，7.62(dd，J＝21.4，6.9Hz，2H)，7.50-7.32(m，2H)，7.26(s，1H)，7.19(s，1H)，6.79(d，J＝7.7Hz，1H)，5.49(s，1H)，4.10(d，J＝13.8Hz，1H)，3.91(s，1H)，3.90(s，1H)，3.71-3.52(m，1H).

Example 19:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (4-methylpiperazin-1-yl) phenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (79mg, 0.37mmol) was dissolved in 8mL of toluene, followed by addition of DPPA (112mg, 0.407mmol), triethylamine (56mg, 0.553mmol) and 4- (4-methylpiperazin-1-yl) aniline (78mg, 0.407 mmol). Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction solution was concentrated, and the residue was purified by flash column chromatography on silica gel (dichloromethane: methanol ═ 10: 1) to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4- (4-methylpiperazin-1-yl) phenyl) urea (15 mg).

LC-MS：403.1，t_R＝1.448min；

¹H NMR(400MHz，MeOD)δ7.93(s，1H)，7.63(d，J＝7.6Hz，1H)，7.56(d，J＝7.6Hz，1H)，7.44(t，J＝7.5Hz，1H)，7.34(td，J＝7.6，1.0Hz，1H)，7.18(dd，J＝9.4，2.8Hz，3H)，6.97-6.83(m，2H)，5.45(dd，J＝6.1，4.6Hz，1H)，4.04(dd，J＝14.3，4.3Hz，1H)，3.53(dd，J＝14.3，6.6Hz，1H)，3.22-3.03(m，4H)，2.77-2.61(m，4H)，2.39(s，3H).

Example 20:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (4-methylpiperazin-1-yl) benzene Yl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (79mg, 0.37mmol) was dissolved in 8mL of toluene, followed by addition of DPPA (112mg, 0.407mmol), triethylamine (56mg, 0.553mmol) and 3-fluoro-4- (4-methylpiperazin-1-yl) aniline (85mg, 0.407 mmol). Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction solution was concentrated, and the residue was purified by flash column chromatography on silica gel (dichloromethane: methanol ═ 10: 1) to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) urea (20 mg).

LC-MS：421.1，t_R＝1.564min；

¹H NMR(400MHz，MeOD)δ7.93(s，1H)，7.64(d，J＝7.6Hz，1H)，7.58(d，J＝7.6Hz，1H)，7.44(t，J＝7.5Hz，1H)，7.35(td，J＝7.6，1.0Hz，1H)，7.26(dd，J＝14.6，2.1Hz，1H)，7.17(s，1H)，6.97-6.86(m，2H)，5.49-5.39(m，1H)，4.07(dd，J＝14.4，4.3Hz，1H)，3.57(dd，J＝14.3，6.4Hz，1H)，3.33(dt，J＝3.3，1.6Hz，2H)，3.05(s，4H)，2.65(s，4H)，2.38(s，3H).

Example 21:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1-benzyl-1H-pyrazol-4-yl) benzene Yl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (79.2mg, 0.37mmol) was dissolved in 8mL of toluene, followed by the addition of DPPA (112mg, 0.407mmol), triethylamine (56mg, 0.555mmol) and 4- (1-benzyl-1H-pyrazol-4-yl) aniline (101mg, 0.407 mmol). Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction was concentrated, and the residue was purified by flash silica gel column chromatography (dichloromethane: methanol 10: 1) to give a white solid (20 mg).

LC-MS：416.2，t_R＝2.124min；

¹H NMR(400MHz，MeOD)δ7.92-7.12(m，17H)，5.45(s，1H)，5.33(s，2H)，4.14-3.96(m，1H)，3.39(dd，J＝14.4，6.6Hz，1H).

Example 22:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea

Preparation of 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea from 2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (300mg, 1.4mmol), 4-bromo-3-fluoroaniline (320mg, 1.684mmol), DPPA (465mg, 1.690mmol) and triethylamine (215mg, 2.125mmol) in toluene (20mL) under nitrogen at reflux for 4 hours, cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography (330 mg).

LC-MS：401.0，t_R＝2.113min；

¹H NMR(400MHz，MeOD)δ7.95(s，1H)，7.65(d，J＝7.6Hz，1H)，7.59(d，J＝7.6Hz，1H)，7.52-7.32(m，4H)，7.18(s，1H)，6.93(dd，J＝8.4，2.0Hz，1H)，5.57-5.44(m，1H)，4.11(dd，J＝14.4，4.2Hz，1H)，3.62(dd，J＝14.4，6.2Hz，1H)；

¹⁹F NMR(400MHz，MeOD)δ-108.8.

Example 23:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1-methyl-1H-pyrazol-4-yl) Phenyl) urea

The preparation method is the same as example 18.

LC-MS：403.1，t_R＝1.920min；

¹H NMR(400MHz，MeOD)δ7.99(s，1H)，7.95-7.93(m，1H)，7.81(s，1H)，7.66(d，J＝7.6Hz，1H)，7.61(d，J＝7.6Hz，1H)，7.55-7.42(m，2H)，7.41-7.34(m，2H)，7.19(s，1H)，7.00(dd，J＝8.5，2.1Hz，1H)，5.57-5.47(m，1H)，4.12(dd，J＝14.4，4.2Hz，1H)，3.94(s，2H)，3.62(dd，J＝14.4，6.2Hz，1H)；

¹⁹F NMR(400MHz，MeOD)δ-115.8(s，1H).

Example 24:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-cyano-3-fluorophenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (79mg, 0.37mmol) was dissolved in 8mL of toluene, followed by addition of DPPA (112mg, 0.407mmol), triethylamine (56mg, 0.553mmol) and 4-amino-2-fluorobenzonitrile (55mg, 0.407 mmol). Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction was concentrated, and the residue was purified by flash column chromatography on silica gel (dichloromethane: methanol ═ 10: 1) to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-cyano-3-fluorophenyl) urea (10 mg).

LC-MS：348.1，t_R＝1.979min；

¹H NMR(400MHz，MeOD)δ7.98(s，1H)，7.72-7.51(m，4H)，7.45(t，J＝7.4Hz，1H)，7.41-7.33(m，1H)，7.19(s，1H)，7.11(dd，J＝8.6，1.9Hz，1H)，5.55-5.45(m，1H)，4.14(dd，J＝14.4，4.1Hz，1H)，3.66(dd，J＝14.4，5.9Hz，1H).

Example 25:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-chloro-4-cyanophenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (79mg, 0.37mmol) was dissolved in 8mL of toluene, followed by addition of DPPA (112mg, 0.407mmol), triethylamine (56mg, 0.553mmol) and 4-amino-2-chlorobenzonitrile (62mg, 0.407 mmol). Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction solution was concentrated, and the residue was purified by flash column chromatography on silica gel (dichloromethane: methanol ═ 10: 1) to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (3-chloro-4-cyanophenyl) urea (10 mg).

LC-MS：364.0，t_R＝2.008min；

¹H NMR(400MHz，MeOD)δ9.16(s，1H)，7.80(ddd，J＝26.8，8.8，3.3Hz，4H)，7.67-7.53(m，3H)，7.26(dd，J＝8.6，1.9Hz，1H)，5.85(s，1H)，4.46-4.36(m，1H)，3.84-3.74(m，1H).

Example 26:

1- (4- (1H-imidazol-1-yl) phenyl) -3- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (100mg, 0.43mmol), Diphenylphosphorylazide (DPPA) (250mg, 0.92mmol), triethylamine (232mg, 2.32mmol), 4- (1H-imidazol-1-yl) aniline (150mg, 0.92mmol) were dissolved in anhydrous toluene (10mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (4- (1H-imidazol-1-yl) phenyl) -3- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (3.4 mg).

LC-MS：371.0，t_R＝1.468min；

¹H NMR(400MHz，MeOD)δ8.05(s，1H)，7.96(s，1H)，7.63(dd，J＝19.9，7.5Hz，2H)，7.48(dt，J＝16.7，5.1Hz，6H)，7.36(t，J＝7.5Hz，1H)，7.16(d，J＝19.2Hz，2H)，5.58-5.45(m，1H)，4.12(dd，J＝14.4，4.2Hz，1H)，3.63(dd，J＝14.4，6.3Hz，1H).

Example 27:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3, 5-difluorophenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (200mg, 0.93mmol), Diphenylphosphorylazide (DPPA) (520mg, 1.86mmol), triethylamine (190mg, 1.86mmol), 4-bromo-3, 5-difluoroaniline (400mg, 1.86mmol) were dissolved in anhydrous toluene (10mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column chromatography to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-bromo-3, 5-difluorophenyl) urea (3.0 mg).

LC-MS：418.2，t_R＝2.259min；

¹H NMR(400MHz，MeOD)δ7.95(s，1H)，7.62(dd，J＝21.3，7.5Hz，2H)，7.41(dt，J＝35.5，7.4Hz，2H)，7.24-7.06(m，3H)，5.58-5.45(m，1H)，4.11(dd，J＝14.3，4.1Hz，1H)，3.63(dd，J＝14.4，6.1Hz，1H).

Example 28:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3, 5-difluoro-4- (1-methyl-1H-pyrazole- 4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3, 5-difluorophenyl) urea (30mg, 0.07mmol), 1-methyl-4- (4, 4, 5, 5-Tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (32mg, 0.15mmol), Pd (dppf) Cl₂(5mg), potassium carbonate (20mg, 0.14mmol) was dissolved in a mixture of dioxane (3mL) and water (1.5mL) and heated at reflux under nitrogen for 2 h. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (10mL) and washed with water (2X 5 mL). The organic phase was dried, filtered and concentrated. Purifying the residue with flash silica gel column to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (2.0 mg).

LC-MS：421.1，t_R＝1.970min；

¹H NMR(400MHz，MeODδ7.96(d，J＝5.4Hz，2H)，7.83(s，1H)，7.62(dd，J＝20.2，7.2Hz，2H)，7.41(dt，J＝34.6，7.3Hz，2H)，7.26-7.01(m，3H)，5.50(s，1H)，4.11(d，J＝11.2Hz，1H)，3.95(s，3H)，3.62(dd，J＝14.3，6.1Hz，1H).

Example 29:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-bromo-4-fluorophenyl) urea

The same procedure as in example 27 was followed.

LC-MS：401.0，t_R＝2.083min；

¹H NMR(400MHz，MeOD)δ7.94(s，1H)，7.72(dd，J＝6.2，2.5Hz，1H)，7.64(d，J＝7.5Hz，1H)，7.58(d，J＝7.6Hz，1H)，7.45(t，J＝7.5Hz，1H)，7.36(t，J＝7.5Hz，1H)，7.23-7.15(m，2H)，7.08(t，J＝8.7Hz，1H)，5.52-5.45(m，1H)，4.09(dd，J＝14.4，4.2Hz，1H)，3.60(dd，J＝14.4，6.2Hz，1H)；

¹⁹F NMR(400MHz，MeOD)δ-118.2.

Example 30:

1- ((5H-imidazole)And [5, 1-a ]]Isoindol-5-yl) methyl) -3- (4-bromo-3-chlorophenyl) urea

The same procedure as in example 27 was followed.

LC-MS：419.0，t_R＝2.252min；

¹H NMR(400MHz，MeOD)δ7.95(s，1H)，7.70(d，J＝2.5Hz，1H)，7.65(d，J＝7.5Hz，1H)，7.60(d，J＝7.7Hz，1H)，7.50(d，J＝8.8Hz，1H)，7.45(t，J＝7.5Hz，1H)，7.36(t，J＝7.1Hz，1H)，7.18(s，1H)，7.09(dd，J＝8.8，2.5Hz，1H)，5.52-5.47(m，1H)，4.11(dd，J＝14.4，4.2Hz，1H)，3.62(dd，J＝14.4，6.2Hz，1H).

Example 31:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-chloro-4- (1-methyl-1H-pyrazol-4-yl) Phenyl) urea

The procedure is as in example 28.

LC-MS：419.0，t_R＝2.416min；

¹H NMR(400MHz，MeOD)δ7.84(d，J＝9.0Hz，2H)，7.64(s，1H)，7.53(d，J＝7.5Hz，1H)，7.51-7.46(m，2H)，7.33(t，J＝7.4Hz，1H)，7.26(dd，J＝14.0，8.0Hz，2H)，7.10-7.04(m，2H)，5.43-5.34(m，1H)，3.99(dd，J＝14.4，4.1Hz，1H)，3.83(s，3H)，3.50(dd，J＝14.3，6.2Hz，1H).

Example 32:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-2-methoxyphenyl) urea

The same procedure as in example 27 was followed.

LC-MS：414.0，t_R＝2.140min；

¹H NMR(400MHz，MeOD)δ7.94(s，1H)，7.89(d，J＝8.7Hz，1H)，7.65(d，J＝7.6Hz，1H)，7.59(d，J＝7.6Hz，1H)，7.45(t，J＝7.5Hz，1H)，7.36(t，J＝7.6Hz，1H)，7.17(s，1H)，7.08(d，J＝2.1Hz，1H)，7.01(dd，J＝8.6，2.1Hz，1H)，5.56-5.42(m，1H)，4.09(dd，J＝14.4，4.2Hz，1H)，3.86(s，3H)，3.62(dd，J＝14.4，6.1Hz，1H).

Example 33:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2-methoxy-4- (1-methyl-1H-pyrazole- 4-yl) phenyl) urea

The preparation method is the same as example 28.

LC-MS：415.1，t_R＝1.888min；

¹H NMR(400MHz，d₆-DMSO)δ8.06(s，2H)，8.00(d，J＝8.3Hz，1H)，7.89(s，1H)，7.80(s，1H)，7.62(d，J＝7.6Hz，1H)，7.56(d，J＝7.5Hz，1H)，7.41(t，J＝7.4Hz，1H)，7.30(t，J＝7.4Hz，1H)，7.17-7.09(m，2H)，7.03(dd，J＝8.4，1.6Hz，1H)，6.98(t，J＝5.9Hz，1H)，5.43-5.40(m，1H)，4.00-3.89(m，1H)，3.86(s，3H)，3.84(s，3H)，3.65-3.47(m，1H).

Example 34:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2-chloro-4-cyanophenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (150mg, 0.7mmol) was dissolved in 10mL of toluene, followed by the addition of DPPA (212mg, 0.77mmol), triethylamine (106mg, 1.05mmol) and 4-amino-3-chlorobenzonitrile (118mg, 0.77 mmol). Heating and refluxing for 2 hours under the protection of nitrogen. The LC-MS detection reaction is complete. The reaction solution was concentrated, and the residue was purified by flash column chromatography on silica gel (dichloromethane: methanol ═ 10: 1) to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (2-chloro-4-cyanophenyl) urea (10 mg).

LC-MS：364.0，t_R＝1.981min；

¹H NMR(400MHz，MeOD)δ9.23(s，1H)，8.27(dd，J＝8.7，2.1Hz，1H)，7.91-7.85(m，1H)，7.76(t，J＝5.0Hz，3H)，7.66-7.51(m，3H)，5.89(t，J＝3.8Hz，1H)，4.49(dd，J＝14.9，3.6Hz，1H)，3.88(dd，J＝14.9，4.3Hz，1H).

Example 35:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-bromophenyl) urea

The preparation method is the same as example 27.

LC-MS：384.0，t_R＝2.081min；

¹H NMR(400MHz，MeOD)δ7.84(s，1H)，7.62-7.50(m，2H)，7.47(d，J＝7.6Hz，1H)，7.33(t，J＝7.4Hz，1H)，7.28-7.18(m，1H)，7.12-6.94(m，4H)，5.43-5.31(m，1H)，3.98(dd，J＝14.4，4.2Hz，1H)，3.49(dd，J＝14.4，6.2Hz，1H).

Example 36:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3- (1-methyl-1H-pyrazol-4-yl) phenyl) Urea

The preparation method is the same as example 28.

LC-MS：385.1，t_R＝1.867min；

¹H NMR(400MHz，d₆-DMSO)δ8.59(s，1H)，8.03(s，1H)，7.90(s，1H)，7.73(s，1H)，7.63(d，J＝7.5Hz，1H)，7.56(dd，J＝6.5，5.1Hz，2H)，7.42(t，J＝7.5Hz，1H)，7.36-7.27(m，1H)，7.23-7.13(m，3H)，7.12-7.05(m，1H)，6.37(t，J＝5.9Hz，1H)，5.52-5.37(m，1H)，3.99-3.87(m，1H)，3.86(s，3H)，3.59-3.46(m，1H).

Example 37:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-cyano-2-fluorophenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (100mg, 0.467mmol), 4-bromo-2-fluoroaniline (76mg, 0.558mmol), DPPA (154mg, 0.560mmol), triethylamine (75mg, 0.741mmol) in toluene (10mL) was refluxed under nitrogen for 4 hours, cooled to room temperature, extracted with water, dichloromethane, dried, and purified by column chromatography to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-cyano-2-fluorophenyl) urea (5.6 mg).

LC-MS：348.1，t_R＝1.943min；

¹H NMR(400MHz，MeOD)δ9.21(s，1H)，8.25(s，1H)，7.68(dd，J＝104.4，48.2Hz，7H)，5.88(s，1H)，4.46(s，2H)，3.86(s，1H).

Example 38:

1- (4- (1H-pyrazol-1-yl) phenyl) -3- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (100mg, 0.467mmol), 4- (1H-pyrazol-1-yl) aniline (90mg0.565mmol), DPPA (155mg, 0.563mmol), triethylamine (75mg, 0.741mmol) in toluene (10mL) was refluxed under nitrogen for 4 hours, cooled to room temperature, added with water, extracted with dichloromethane, dried, and purified by column chromatography to obtain 1- (4- (1H-pyrazol-1-yl) phenyl) -3- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (21 mg).

LC-MS：371.0，t_R＝1.902min；

¹H NMR(400MHz，MeOD)δ8.13(s，2H)，7.51(dd，J＝113.4，52.2Hz，10H)，6.51(s，1H)，5.55(s，1H)，4.14(s，1H)，3.65(s，1H).

Example 39:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-2, 5-difluorophenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (200mg, 0.93mmol), Diphenylphosphorylazide (DPPA) (520mg, 1.86mmol), triethylamine (190mg, 1.86mmol), 4-bromo-2, 5-difluoroaniline (400mg, 1.86mmol) were dissolved in anhydrous toluene (10mL) and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column chromatography to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (4-bromo-2, 5-difluorophenyl) urea (3.0 mg).

LC-MS：421.0，t_R＝2.221min；

¹H NMR(400MHz，MeOD)δ8.04(dd，J＝11.1，7.0Hz，1H)，7.95(s，1H)，7.62(dd，J＝21.4，7.6Hz，2H)，7.45(t，J＝7.4Hz，1H)，7.38(dt，J＝14.1，7.0Hz，2H)，7.17(s，1H)，5.50(t，J＝4.7Hz，1H)，4.15(dd，J＝14.4，4.1Hz，1H)，3.70(dd，J＝14.5，5.6Hz，1H).

Example 40:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-methyl-1H-pyrazole- 4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-2, 5-difluorophenyl) urea (30mg, 0.07mmol), 1-methyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (32mg, 0.15mmol), Pd (dppf) Cl₂(5mg), potassium carbonate (20mg, 0.14mmol) was dissolved in dioxane (3mL), water (0.5mL) and heated under reflux for 2h under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (10mL) and washed with water (2X 5 mL). The organic phase was dried, filtered and concentrated. Purifying the residue with flash silica gel column to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (2.0 mg).

LC-MS：421.0，t_R＝1.963min；

¹H NMR(400MHz，MeOD)δ7.96(s，2H)，7.91(dd，J＝13.1，6.9Hz，1H)，7.83(s，1H)，7.63(dd，J＝20.3，7.5Hz，2H)，7.45(t，J＝7.5Hz，1H)，7.38(dd，J＝16.0，7.7Hz，2H)，7.18(s，1H)，5.51(s，1H)，4.15(dd，J＝14.7，4.0Hz，1H)，3.94(s，3H)，3.69(dd，J＝14.3，5.9Hz，1H).

Examples 41 and 42:

(R) -1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-methyl-1H-pyridine) Oxazol-4-yl) phenyl) urea and (S) -1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1- methyl-1H-pyrazol-4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (420mg) was subjected to chiral resolution to give (R-1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (176mg) and (S) -1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) urea (202 mg). The resolution conditions were as follows:

example 43:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3- (trifluoromethyl) phenyl) urea

The preparation method is the same as example 39.

LC-MS：325.0，t_R＝2.376min；

¹H NMR(400MHz，MeOD)δ8.21(s，1H)，7.82(d，J＝2.4Hz，1H)，7.63(d，J＝8.4Hz，2H)，7.52(d，J＝8.8Hz，1H)，7.45(dt，J＝14.9，7.2Hz，2H)，7.37(dd，J＝8.7，2.6Hz，1H)，7.26(s，1H)，5.59-5.49(m，1H)，4.21(dd，J＝14.4，4.0Hz，1H)，3.64(dd，J＝14.5，5.8Hz，1H)；

¹⁹F NMR(400MHz，MeOD)δ-64.1，-76.9。

Example 44:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-fluoro-3- (1-methyl-1)H-pyrazol-4-yl) Phenyl) urea

The preparation method is the same as example 40.

LC-MS：403.1，t_R＝2.536min；

¹H NMR(400MHz，DMSO)δ8.61(s，1H)，8.05(d，J＝1.9Hz，1H)，7.91(s，1H)，7.74(s，1H)，7.66-7.60(m，2H)，7.56(d，J＝7.5Hz，1H)，7.42(t，J＝7.4Hz，1H)，7.31(t，J＝7.5Hz，1H)，7.23-7.04(m，3H)，6.37(t，J＝5.8Hz，1H)，5.515.37(m，1H)，3.93-3.84(m，1H)，3.86(s，3H)，3.59-3.46(m，1H)；

¹⁹F NMR(400MHz，DMSO)δ-124.1.

Example 45:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1-ethyl-1H-pyrazol-4-yl) Phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea (100mg, 0.249mmol), 1-ethyl-1H-pyrazole-4-boronic acid pinacol ester (110mg, 0.495mmol), Pd (dppf) Cl₂(91mg, 0.124mmol) and potassium carbonate (69mg, 0.499mmol) were stirred in 1, 4-dioxane/water (10mL/2mL) and the temperature was raised to 110 ℃ for 1 hour by microwave reaction under nitrogen. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1-ethyl-1H-pyrazol-4-yl) phenyl) urea (33.4 mg).

LC-MS：417.1，t_R＝1.998min；

¹H NMR(400MHz，MeOD)δ7.96(s，2H)，7.81(s，1H)，7.61(dd，J＝20.9，7.5Hz，2H)，7.54-7.30(m，4H)，7.17(s，1H)，6.99(dd，J＝8.5，1.6Hz，1H)，5.47(s，1H)，4.22(q，J＝7.3Hz，2H)，4.09(dd，J＝14.4，3.8Hz，1H)，3.59(dd，J＝14.3，6.0Hz，1H)，1.48(t，J＝7.3Hz，3H).

Example 46:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1H-pyrazol-4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea (100mg, 0.249mmol), 1H-pyrazole-4-boronic acid pinacol ester (97mg, 0.5mmol), Pd (dppf) Cl₂(91mg, 0.124mmol) and potassium carbonate (69mg, 0.499mmol) were stirred in 1, 4-dioxane/water (10mL/2mL) and the temperature was raised to 110 ℃ for 1 hour by microwave reaction under nitrogen. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1H-pyrazol-4-yl) phenyl) urea (16.7 mg).

LC-MS：389.2，t_R＝1.793min；

¹H NMR(400MHz，MeOD)δ7.96(d，J＝13.4Hz，3H)，7.76-7.30(m，6H)，7.19(s，1H)，7.01(d，J＝8.0Hz，1H)，5.51(s，1H)，4.11(d，J＝11.6Hz，1H)，3.62(dd，J＝14.5，5.7Hz，1H).

Example 47:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (2-methoxyethyl) -1H- Pyrazol-4-yl) phenyl) urea

The first step is as follows: preparation of 1- (2-methoxyethyl) -1H-pyrazole-4-boronic acid pinacol ester

1H-pyrazole-4-boronic acid pinacol ester (1.0g, 5.155mmol), 2-bromoethyl methyl ether (0.788g, 5.669mmol) and cesium carbonate (5.04g, 15.469mmol) were dissolved in acetonitrile (20mL), stirred overnight at 50 ℃ under nitrogen protection, filtered, and concentrated to dryness to give 1- (2-methoxyethyl) -1H-pyrazole-4-boronic acid pinacol ester (1.310 g).

LC-MS：253.1，t_R＝2.373min.

The second step is that: 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (2-methoxyethyl) ethyl ester Preparation of 1H-pyrazol-4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea (129mg, 0.322mmol), 1- (2-methoxyethyl) -1H-pyrazole-4-boronic acid pinacol ester (162mg, 0.643mmol), Pd (dppf) Cl₂(118mg, 0.161mmol) and potassium carbonate (89mg, 0.644mmol) were stirred in 1, 4-dioxane/water (10mL/2mL) and the reaction was microwaved to 110 ℃ for 1 hour under nitrogen. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (2-methoxyethyl) -1H-pyrazol-4-yl) phenyl) urea (53.0 mg).

LC-MS：447.1，t_R＝1.912min；

¹H NMR(400MHz，CDCl₃)δ9.75(s，1H)，8.10(s，1H)，7.79(d，J＝4.2Hz，2H)，7.64-7.31(m，7H)，7.21(s，1H)，7.09(d，J＝8.1Hz，1H)，6.68(s，1H)，5.60(d，J＝7.2Hz，1H)，4.30(s，3H)，3.78(d，J＝5.0Hz，3H)，3.32(d，J＝14.5Hz，3H).

Example 48:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1-isopropyl-1H-pyrazole-4-carboxylic acid) Yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea (120mg, 0.299mmol), 1-isopropyl-1H-pyrazole-4-boronic acid pinacol ester (141mg, 0.597mmol), Pd (dppf) Cl₂(109mg, 0.149mmol) and potassium carbonate (83mg, 0.601mmol) were stirred in 1, 4-dioxane/water (10mL/2mL) and the reaction was microwaved to 110 ℃ for 1 hour under nitrogen. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1-isopropyl-1H-pyrazol-4-yl) phenyl) urea (14.0 mg).

LC-MS：431.2，t_R＝2.101min；

¹H NMR(400MHz，MeOD)δ9.00(s，1H)，7.98(s，1H)，7.92-7.07(m，8H)，6.95(d，J＝7.9Hz，1H)，6.40(s，1H)，5.80(s，1H)，4.57(s，1H)，4.36(d，J＝15.1Hz，1H)，3.77(s，1H)，1.31(s，6H).

Example 49:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1-isobutyl-1H-pyrazole-4-one) Yl) phenyl) urea

1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (3-fluoro-4- (1H-pyrazol-4-yl) phenyl) urea (12mg, 0.031mmol), isobutyl bromide (8.5mg, 0.062mmol) and cesium carbonate (30mg, 0.092mmol) were stirred in 2mL acetonitrile at 50 ℃ under nitrogen for overnight stirring, filtered, and the filtrate was purified by column chromatography to give 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (3-fluoro-4- (1-isobutyl-1H-pyrazol-4-yl) phenyl) urea (7.3 mg).

LC-MS：445.1，t_R＝2.219min；

¹H NMR(400MHz，MeOD)δ7.86(s，2H)，7.73(s，1H)，7.52(dd，J＝18.2，7.6Hz，2H)，7.45-7.21(m，4H)，7.07(s，1H)，6.93(dd，J＝8.4，2.0Hz，1H)，5.47-5.33(m，1H)，3.98(dd，J＝14.3，4.3Hz，1H)，3.89(d，J＝7.3Hz，2H)，3.50(dd，J＝14.3，6.5Hz，1H)，2.16-2.07(m，1H)，0.83(t，J＝5.8Hz，6H).

Example 50:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1-isobutyl-1H-pyri-dine Oxazol-4-yl) phenyl) urea

The first step is as follows: preparation of 5-chloro-2-fluoro-4- (1-isobutyl-1H-pyrazol-4-yl) aniline

4-bromo-5-chloro-2-fluoroaniline (100mg, 0.446mmol), 1-isobutyl-1H-pyrazole-4-boronic acid pinacol ester (223mg, 0.891mmol), Pd (dppf) Cl₂(163mg, 0.223mmol) and potassium carbonate (123mg, 0.890mmol) were stirred in 1, 4-dioxane/water (10mL/2mL) and the temperature was raised to 110 ℃ under nitrogen protection for 1 hour of microwave reaction. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 5-chloro-2-fluoro-4- (1-isobutyl-1H-pyrazol-4-yl) aniline (67.0 mg).

LC-MS：268.1，270.1，t_R＝2.712min.

The second step is that: 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1-isobutyl) ether Preparation of 1H-pyrazol-4-yl) phenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (45mg, 0.209mmol), 5-chloro-2-fluoro-4- (1-isobutyl-1H-pyrazol-4-yl) aniline (67mg, 0.250mmol), DPPA (69mg, 0.251mmol) and triethylamine (32mg, 316mol) were refluxed under the protection of toluene (10mL) for 5 hours, cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1-isobutyl-1H-pyrazol-4-yl) phenyl) urea (9 mg).

LC-MS：479.2.0，481.1，t_R＝2.310min；

¹H NMR(400MHz，MeOD)δ8.11(s，1H)，7.91(d，J＝29.2Hz，2H)，7.73(s，1H)，7.53(d，J＝19.0Hz，2H)，7.27(dd，J＝40.6，22.5Hz，3H)，7.08(s，1H)，5.41(s，1H)，4.05(d，J＝11.8Hz，1H)，3.91(s，2H)，3.59(d，J＝9.4Hz，1H)，2.12(s，1H)，0.85(d，J＝3.7Hz，6H).

Example 51:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1-cyclobutylmethyl- 1H-pyrazol-4-yl) phenyl) urea

The first step is as follows: preparation of 1-cyclobutylmethyl-1H-pyrazole-4-boronic acid pinacol ester

1H-pyrazole-4-boronic acid pinacol ester (500mg, 2.577mmol), cyclobutylmethyl bromide (768mg, 5.153mmol) and cesium carbonate (2.520g, 7.734mmol) were dissolved in acetonitrile (10mL), stirred overnight at 50 ℃ under nitrogen protection, filtered, and concentrated to dryness to give 1-cyclobutylmethyl-1H-pyrazole-4-boronic acid pinacol ester (405 mg).

The second step is that: preparation of 5-chloro-2-fluoro-4- (1-cyclobutylmethyl-1H-pyrazol-4-yl) aniline

4-bromo-5-chloro-2-fluoroaniline (86mg, 0.383mmol), 1-cyclobutylmethyl-1H-pyrazole-4-boronic acid pinacol ester (200mg, 0.763mmol), Pd (dppf) Cl₂(140mg, 0.191mmol) and potassium carbonate (105mg, 0.760mmol) were stirred in 1, 4-dioxane/water (10mL/2mL) and the temperature was raised to 110 ℃ under nitrogen for 1 hour of microwave reaction. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 5-chloro-2-fluoro-4- (1-cyclobutylmethyl-1H-pyrazol-4-yl) aniline (47.0 mg).

LC-MS：280.0，282.1，t_R＝2.787min.

The third step: 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1-cyclobutyl) Preparation of methyl-1H-pyrazol-4-yl) phenyl) urea

2- (5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (30mg, 0.140mmol), 5-chloro-2-fluoro-4- (1-cyclobutylmethyl-1H-pyrazol-4-yl) aniline (47mg, 0.168mmol), DPPA (46mg, 0.167mmol) and triethylamine (21mg, 0.208mmol) were refluxed in toluene (10mL) under nitrogen for 5 hours, cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1-cyclobutylmethyl-1H-pyrazol-4-yl) phenyl) urea (1.6 mg).

LC-MS：491.2，493.1，t_R＝2.430min；

¹H NMR(400MHz，MeOD)δ8.19(d，J＝7.6Hz，1H)，8.03(s，2H)，7.80(s，1H)，7.64(dd，J＝23.4，7.5Hz，2H)，7.43(dt，J＝30.5，7.4Hz，2H)，7.30(d，J＝12.2Hz，2H)，5.53(s，1H)，4.20(d，J＝7.3Hz，2H)，3.71(dd，J＝14.5，5.6Hz，1H)，2.87(dt，J＝15.0，7.5Hz，1H)，2.24-2.17(m，1H)，2.09(dd，J＝16.8，8.2Hz，3H)，1.89(ddd，J＝25.6，14.0，7.8Hz，3H).

Examples 52 and 53: 2- (4- (4- (3- (5H-imidazo [5, 1-a))]Isoindol-5-yl) methyl) ureido) -2-fluoro Phenyl) -1H-pyrazol-1-yl) ethyl acetate and 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro- 4- (1- (2-hydroxyethyl) -1H-pyrazol-4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (3-fluoro-4- (1H-pyrazol-4-yl) phenyl) urea (74mg), 2-bromoethyl acetate (34mg, 0.204mmol) and cesium carbonate (167mg, 0.513mmol) were stirred in acetonitrile (10mL) at 50 ℃ under nitrogen protection overnight, filtered, and the filtrate was purified by column chromatography to give 2- (4- (4- (3- (5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) ureido) -2-fluorophenyl) -1H-pyrazol-1-yl) ethyl acetate (5.2mg) and 1- ((5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) -3- (3- Fluoro-4- (1- (2-hydroxyethyl) -1H-pyrazol-4-yl) phenyl) urea (3.1 mg).

Example 52:

LC-MS：475.1，t_R＝1.936min；

¹H NMR(400MHz，CDCl₃)δ9.35(s，1H)，9.02(s，1H)，8.00-7.31(m，8H)，7.19(s，1H)，5.65(s，1H)，5.35(s，1H)，4.41(d，J＝29.1Hz，3H)，3.69(d，J＝46.1Hz，4H)，2.04(s，3H).

example 53:

LC-MS：433.1，t_R＝2.780min；

¹H NMR(400MHz，MeOD)δ8.00(s，2H)，7.85(s，1H)，7.65(d，J＝21.0Hz，2H)，7.44(d，J＝35.4Hz，5H)，7.00(s，1H)，5.52(s，1H)，4.27(s，2H)，4.14(d，J＝11.6Hz，1H)，3.92(s，2H)，3.65(s，1H).

example 54:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-isobutyl-1H-pyrane Oxazol-4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-2, 5-difluorophenyl) urea (30mg, 0.07mmol), 1-isobutyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (32mg, 0.15mmol), Pd (dppf) Cl₂(5mg) and potassium carbonate (20mg, 0.14mmol) were dissolved in dioxane (3mL) and water (0.5mL) and heated at reflux under nitrogen for 2 h. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was washed with dichloromethane (10mL) and water (2X 5 mL). The organic phase was dried, filtered and concentrated. Purifying the residue with flash silica gel column to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-isobutyl-1H-pyrazol-4-yl) phenyl) urea (2.2 mg).

LC-MS：463.1，t_R＝2.263min；

¹H NMR(400MHz，MeOD)δ8.01-7.73(m，1H)，7.54(dd，J＝21.2，7.5Hz，1H)，7.43-7.24(m，1H)，7.11(s，1H)，5.41(s，1H)，4.06(dd，J＝14.3，3.7Hz，1H)，3.90(d，J＝7.3Hz，1H)，3.60(dd，J＝14.3，5.6Hz，1H)，2.12(dt，J＝13.5，6.8Hz，1H)，0.83(t，J＝8.5Hz，2H).

Example 55:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (2- (pyrrolidin-1-yl) ethyl) acetate Yl) -1H-pyrazol-4-yl) phenyl) urea

The procedure is as in example 48.

LC-MS：486.1，t_R＝2.153min；

¹H NMR(400MHz，MeOD)δ7.96(d，J＝1.4Hz，1H)，7.88(s，1H)，7.81(s，1H)，7.56(d，J＝7.5Hz，1H)，7.51(d，J＝7.5Hz，1H)，7.42(t，J＝8.6Hz，1H)，7.36(t，J＝7.5Hz，1H)，7.32(d，J＝2.0Hz，1H)，7.31-7.25(m，2H)，7.10(s，1H)，6.92(dd，J＝8.5，2.0Hz，1H)，5.46-5.37(m，1H)，4.38(t，J＝6.3Hz，2H)，4.03(dd，J＝14.4，4.2Hz，1H)，3.53(dd，J＝14.4，6.2Hz，1H)，3.34-3.24(m，2H)，1.90-1.80(m，4H)，1.30-1.20(m，4H)；

¹⁹F NMR(400MHz，MeOD)δ-115.7.

Example 56:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (2-morpholinoethyl) -1H- Pyrazol-4-yl) phenyl) urea

The procedure is as in example 48.

LC-MS：502.1，t_R＝2.014min；

¹H NMR(400MHz，MeOD)δ9.18(s，1H)，8.08(s，1H)，7.93(s，1H)，7.90-7.82(m，1H)，7.75(d，J＝8.0Hz，2H)，7.60(dd，J＝5.5，3.2Hz，2H)，7.50(t，J＝8.6Hz，1H)，7.38(dd，J＝13.7，1.5Hz，1H)，6.98(d，J＝8.4Hz，1H)，5.86(t，J＝4.0Hz，1H)，4.68(t，J＝5.8Hz，2H)，4.37(dd，J＝14.8，3.5Hz，1H)，4.20-3.80(m，4H)，3.82-3.69(m，3H)，3.66-3.31(m，4H)；

¹⁹F NMR(400MHz，MeOD)δ-115.6.

Example 57:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1- (cyclopropylmethyl) -1H-pyrazole-4-carboxylic acid Phenyl) -3-fluorophenyl) urea

The procedure is as in example 48.

LC-MS：443.1，t_R＝2.067min；

¹H NMR(400MHz，MeOD)δ7.93(d，J＝1.4Hz，1H)，7.87(s，1H)，7.74(s，1H)，7.56(d，J＝7.5Hz，1H)，7.51(d，J＝7.6Hz，1H)，7.42(t，J＝8.6Hz，1H)，7.36(t，J＝7.5Hz，1H)，7.29(dt，J＝14.4，5.0Hz，2H)，7.09(s，1H)，6.91(dd，J＝8.5，2.0Hz，1H)，5.47-5.38(m，1H)，4.02(dd，J＝14.4，4.2Hz，1H)，3.95(d，J＝7.1Hz，2H)，3.53(dd，J＝14.4，6.2Hz，1H)，1.25(ddd，J＝12.6，7.6，4.8Hz，1H)，0.62-0.49(m，2H)，0.35(q，J＝4.8Hz，2H)；

¹⁹F NMR(400MHz，MeOD)δ-115.8.

Examples 58 and 59:

2- (4- (4- (3 ((5H-imidazo [5, 1-a))]Isoindol-5-yl) methyl) ureido) -2-fluorophenyl) -1H-pyrazole- 1-yl) acetic acid ethyl ester and 2- (4- (4- (3 ((5H-imidazo [5, 1-a))]Isoindol-5-yl) methyl) ureido) -2-fluorophenyl- 1H-pyrazol-1-yl) acetic acid

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea (120mg, 0.299mmol), ethyl 1-acetate-1H-pyrazole-4-boronic acid pinacol ester (191mg), Pd (dppf) Cl₂(109mg, 0.149mmol) and potassium carbonate (83mg, 0.601mmol) were stirred in 1, 4-dioxane/water (10mL/2mL) and the reaction was microwaved to 110 ℃ for 1 hour under nitrogen. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatography to obtain 2- (4- (4- (3 ((5H-imidazo [5, 1-a))]Isoindol-5-yl) methyl) ureido) -2-fluorophenyl) -1H-pyrazol-1-yl acetate (12.3mg) and 2- (4- (4- (3 ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) ureido) -2-fluorophenyl) -1H-pyrazol-1-yl) acetic acid (3.8 mg).

Example 58:

LC-MS：475.1，t_R＝2.030min；

¹H NMR(400MHz，MeOD)δ7.87(d，J＝1.5Hz，2H)，7.74(s，1H)，7.51(d，J＝7.5Hz，1H)，7.45(d，J＝7.5Hz，1H)，7.37(t，J＝8.6Hz，1H)，7.30(dd，J＝14.0，6.5Hz，2H)，7.25-7.18(m，1H)，6.86(d，J＝8.3Hz，1H)，5.34(s，1H)，4.91(s，2H)，4.10(q，J＝7.1Hz，2H)，3.97(d，J＝11.5Hz，1H)，3.47(dd，J＝14.4，6.1Hz，1H)，1.14(d，J＝7.2Hz，3H)；

¹⁹F NMR(400MHz，MeOD)δ-115.6.

example 59:

LC-MS：447.1，t_R＝1.770min；

¹H NMR(400MHz，d₆-DMSO)δ8.97(s，1H)，8.79(s，1H)，8.04(d，J＝1.7Hz，1H)，7.90-7.75(m，2H)，7.70(d，J＝9.6Hz，2H)，7.52(dt，J＝13.9，6.3Hz，3H)，7.43(dd，J＝14.0，1.7Hz，1H)，6.97(dd，J＝8.5，1.7Hz，1H)，6.44(t，J＝6.0Hz，1H)，5.74(s，1H)，4.99(s，2H)，4.21-4.07(m，1H)，3.71(dt，J＝13.9，5.7Hz，1H)；

¹⁹F NMR(400MHz，d6-DMSO)δ-114.0.

example 60:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1- (cyclobutylmethyl) -1H-pyrazole-4-one Phenyl) -3-fluorophenyl) urea

Compound 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) (110mg, 0.274mmol) was dissolved in 10mL of 1, 4-dioxane and 2mL of water, and 1- (cyclobutylmethyl) -4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (86mg, 0.329mmol), sodium carbonate (87mg, 0.823mmol) and Pd (dppf) Cl were added₂(20mg, 0.027 mmol). The reaction was carried out at 104 ℃ for 1 hour under nitrogen protection. The LC-MS detection reaction is complete. The reaction was concentrated, and the residue was purified by flash silica gel column chromatography (dichloromethane: methanol 10: 1) to give 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1- (cyclobutylmethyl) -1H-pyrazol-4-yl) -3-fluorophenyl) urea (5.7 mg).

LC-MS：457.1，t_R＝2.237min；

¹H NMR(400MHz，MeOD)δ7.95(d，J＝6.4Hz，2H)，7.81(s，1H)，7.62(dd，J＝20.2，7.5Hz，2H)，7.54-7.29(m，4H)，7.18(s，1H)，7.01(d，J＝8.4Hz，1H)，5.53-5.42(m，1H)，4.19(d，J＝7.3Hz，2H)，4.10(dd，J＝14.4，4.2Hz，1H)，3.60(dd，J＝14.3，6.3Hz，1H)，2.86(dt，J＝15.2，7.6Hz，1H)，2.15-2.02(m，2H)，2.00-1.67(m，4H).

Example 61:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (tetrahydro-2H-pyran-4-) Yl) -1H-pyrazol-4-yl) phenyl) urea

1- ((5H-miami)Azolo [5, 1-a ]]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea (90mg, 0.224mmol) was dissolved in 10mL of 1, 4-dioxane and 2mL of water, followed by the addition of 1- (tetrahydro-2H-pyran-4-yl) -4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (75mg, 0.269mmol), sodium carbonate (71mg, 0.673mmol) and Pd (dppf) Cl₂(16mg, 0.022 mmol). The reaction was carried out at 104 ℃ for 1 hour under nitrogen protection. The LC-MS detection reaction is complete. The reaction was concentrated, and the residue was purified by flash silica gel column chromatography (dichloromethane: methanol 10: 1) to give 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) phenyl) urea (5.8 mg).

LC-MS：473.2，t_R＝1.943min；

¹H NMR(400MHz，MeOD)δ8.05(d，J＝12.8Hz，2H)，7.85(s，1H)，7.73-7.34(m，7H)，7.23(s，1H)，7.00(dd，J＝8.5，2.0Hz，1H)，5.61-5.43(m，1H)，4.45(ddd，J＝15.7，10.9，4.6Hz，1H)，4.22-3.98(m，3H)，3.61(ddd，J＝13.7，11.2，5.0Hz，3H)，2.22-1.95(m，4H).

Example 62:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1, 3, 5-trimethyl-1H- Pyrazol-4-yl) phenyl) urea

The preparation method is the same as that of example 54.

LC-MS：449.2，t_R＝2.14min；

¹H NMR(400MHz，MeOD)δ7.88-7.78(m，2H)，7.51(dd，J＝19.5，7.5Hz，2H)，7.34(t，J＝7.5Hz，1H)，7.29-7.22(m，1H)，7.06(s，1H)，6.86(dd，J＝11.4，6.7Hz，1H)，5.40(t，J＝4.8Hz，1H)，4.04(dd，J＝14.4，4.1Hz，1H)，3.65(s，3H)，3.58(dd，J＝14.4，5.7Hz，1H)，2.06(d，J＝7.1Hz，3H)，1.99(s，3H).

Example 63:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (2- (2-carbonylpyrrolidine-) 1-yl) ethyl) -1H-pyrazol-4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-3-fluorophenyl) urea (100mg, 0.249mmol) was dissolved in 10mL of 1, 4-dioxane and 2mL of water, and 1- (2- (4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazol-1-yl) ethyl) pyrrolidin-2-one (91mg, 0.299mmol), sodium carbonate (79mg, 0.748mmol) and Pd (dppf) Cl were added₂(18mg, 0.025 mmol). The reaction was carried out under nitrogen at 104 ℃ for 1 hour. The LC-MS detection reaction is complete. The reaction was concentrated, and the residue was purified by flash silica gel column chromatography (dichloromethane: methanol 10: 1) to give 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (3-fluoro-4- (1- (2- (2-carbonylpyrrolidin-1-yl) ethyl) -1H-pyrazol-4-yl) phenyl) urea (8.5 mg).

LC-MS：500.2，t_R＝1.823min；

¹H NMR(400MHz，MeOD)δ8.06(s，1H)，7.97(d，J＝1.6Hz，1H)，7.85(s，1H)，7.63(dd，J＝23.0，7.5Hz，2H)，7.47(dt，J＝12.1，8.0Hz，2H)，7.42-7.34(m，2H)，7.22(s，1H)，6.99(dd，J＝8.5，2.0Hz，1H)，5.52(t，J＝5.0Hz，1H)，4.36(t，J＝5.8Hz，2H)，4.12(dd，J＝14.4，4.1Hz，1H)，3.70(t，J＝5.8Hz，2H)，3.62(dd，J＝14.4，6.2Hz，1H)，3.21(t，J＝7.0Hz，2H)，2.31(t，J＝8.1Hz，2H)，2.01-1.90(m，2H).

Example 64:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1, 3, 5-trimethyl- 1H-pyrazol-4-yl) phenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-5-chloro-2-fluorophenyl) urea (70mg, 0.161mmol) was dissolved in 10mL1, 4-dioxane and 2mL water, followed by the addition of 1, 3, 5-trimethyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (57mg, 0.241mmol), sodium carbonate (51mg, 0.482mmol) and Pd (dppf) Cl₂(12mg, 0.016 mmol). The reaction was carried out under nitrogen at 104 ℃ for 1 hour. The LC-MS detection reaction is complete. The reaction was concentrated, and the residue was purified by flash silica gel column chromatography (dichloromethane: methanol 10: 1) to give 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1, 3, 5-trimethyl-1H-pyrazol-4-yl) phenyl) urea (3.5 mg).

LC-MS：465.1，t_R＝2.007min；

¹H NMR(400MHz，MeOD)δ8.24(d，J＝7.6Hz，1H)，7.97(s，1H)，7.63(dd，J＝19.5，7.5Hz，2H)，7.46(t，J＝7.5Hz，1H)，7.38(t，J＝7.5Hz，1H)，7.19(s，1H)，7.00(d，J＝11.5Hz，1H)，5.52(t，J＝4.7Hz，1H)，4.17(dt，J＝14.4，3.7Hz，1H)，3.75(d，J＝13.0Hz，3H)，3.71(dd，J＝14.4，5.6Hz，1H)，2.09(d，J＝24.4Hz，6H).

Example 65:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1-isobutyl-3, 5-di methyl-1H-pyrazol-4-yl) phenyl) urea

The preparation method is the same as example 64.

LC-MS：507.1，t_R＝2.35min；

¹H NMR(400MHz，MeOD)δ8.24(d，J＝7.5Hz，1H)，7.98(s，1H)，7.63(dd，J＝19.7，7.6Hz，2H)，7.46(t，J＝7.5Hz，1H)，7.38(t，J＝7.5Hz，1H)，7.19(s，1H)，7.01(d，J＝11.5Hz，1H)，5.52(t，J＝4.7Hz，1H)，4.17(dt，J＝14.4，4.0Hz，1H)，3.85(dd，J＝7.5，1.0Hz，2H)，3.71(dd，J＝14.4，5.6Hz，1H)，2.21(dt，J＝13.7，6.9Hz，1H)，2.09(d，J＝17.2Hz，6H)，0.94(t，J＝6.6Hz，6H).

Example 66:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-4- (3, 5-dimethyl-1H-pyrazole- 4-yl) -2-fluorophenyl) urea

The preparation method is the same as example 64.

LC-MS：451.1，t_R＝1.954min；

¹H NMR(400MHz，MeOD)δ8.23(d，J＝7.6Hz，1H)，8.02(s，1H)，7.64(dd，J＝20.5，7.6Hz，2H)，7.47(t，J＝7.5Hz，1H)，7.42-7.36(m，1H)，7.21(s，1H)，7.01(d，J＝11.5Hz，1H)，5.53(t，J＝4.7Hz，1H)，4.18(dd，J＝14.4，4.1Hz，1H)，3.71(dd，J＝14.4，5.6Hz，1H)，2.11(s，6H).

Example 67:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1- (cyclopropylmethyl) -3, 5-dimethyl 1H-pyrazol-4-yl) -2, 5-difluorophenyl) urea

The preparation method is the same as that of example 54.

LC-MS：489.1，t_R＝2.21min；

¹H NMR(400MHz，MeOD)δ7.86(s，1H)，7.82(dd，J＝11.9，7.0Hz，1H)，7.54(d，J＝7.5Hz，1H)，7.49(d，J＝7.6Hz，1H)，7.34(t，J＝7.4Hz，1H)，7.26(t，J＝7.6Hz，1H)，7.07(d，J＝4.8Hz，1H)，6.87(dd，J＝11.4，6.7Hz，1H)，5.40(t，J＝4.7Hz，1H)，4.05(dd，J＝14.4，4.1Hz，1H)，3.84(d，J＝6.8Hz，2H)，3.59(dd，J＝14.5，5.7Hz，1H)，2.08(s，3H)，2.01(s，3H)，1.31-1.07(m，6H)，0.47(q，J＝5.8Hz，2H)，0.29(q，J＝4.9Hz，2H)；

¹⁹F NMR(400MHz，MeOD)δ-119.43，-138.60.

Example 68:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-4- (1- (cyclopropylmethyl) -3, 5- dimethyl-1H-pyrazol-4-yl) -2-fluorophenyl) urea

The preparation method is the same as example 64.

LC-MS：505.1，t_R＝2.24min；

¹H NMR(400MHz，MeOD)δ8.24(d，J＝7.6Hz，1H)，7.98(s，1H)，7.66(d，J＝7.5Hz，1H)，7.61(d，J＝7.4Hz，1H)，7.46(t，J＝7.5Hz，1H)，7.38(t，J＝7.5Hz，1H)，7.19(s，1H)，7.01(d，J＝11.5Hz，1H)，5.52(t，J＝4.8Hz，1H)，4.17(dt，J＝14.4，4.2Hz，1H)，3.95(dt，J＝9.8，4.9Hz，2H)，3.71(dd，J＝14.4，5.6Hz，2H)，2.14(s，3H)，2.08(s，3H)，0.59(d，J＝8.1Hz，2H)，0.45-0.35(m，2H)；

¹⁹F NMR(400MHz，MeOD)δ-134.87.

Example 69:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1-isobutyl-3, 5-di methyl-1H-pyrazol-4-yl) phenyl) urea

The preparation method is the same as that of example 54.

LC-MS：491.2，t_R＝2.301min；

¹H NMR(400MHz，MeOD)δ8.01(s，1H)，7.94(dd，J＝11.9，7.0Hz，1H)，7.64(dd，J＝20.5，7.6Hz，2H)，7.46(t，J＝7.5Hz，1H)，7.38(t，J＝7.6Hz，1H)，7.20(s，1H)，6.99(dd，J＝11.4，6.7Hz，1H)，5.53(t，J＝4.7Hz，1H)，4.18(dd，J＝14.4，4.1Hz，1H)，3.86(d，J＝7.5Hz，2H)，3.71(dd，J＝14.4，5.7Hz，1H)，2.68(s，1H)，2.17(s，3H)，2.13(s，3H)，0.94(d，J＝6.7Hz，6H).

Example 70:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-2-fluoro-4- (1- (2-methoxyethyl) ethyl Yl) -3, 5-dimethyl-1H-pyrazol-4-yl) phenyl) urea

The preparation method is the same as example 64.

LC-MS：509.2，t_R＝2.086min；

¹H NMR(400MHz，MeOD)δ8.23(d，J＝7.6Hz，1H)，8.00(s，1H)，7.64(dd，J＝20.2，7.6Hz，2H)，7.46(t，J＝7.4Hz，1H)，7.38(t，J＝7.1Hz，1H)，7.20(s，1H)，7.00(d，J＝11.5Hz，1H)，5.53(t，J＝4.8Hz，1H)，4.28-4.12(m，3H)，3.73(qd，J＝10.3，5.3Hz，3H)，3.31(s，3H)，2.10(d，J＝23.8Hz，6H).

Example 71:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (2, 5-difluoro-4- (1- (2-methoxyethyl) ethyl Yl) -3, 5-dimethyl-1H-pyrazol-4-yl) phenyl) urea

The preparation method is the same as that of example 54.

LC-MS：493.1，t_R＝2.010min；

¹H NMR(400MHz，MeOD)δ8.05-7.86(m，2H)，7.63(dd，J＝20.3，7.5Hz，2H)，7.49-7.33(m，2H)，7.19(s，1H)，6.98(dd，J＝11.4，6.7Hz，1H)，5.52(t，J＝4.8Hz，1H)，4.30-4.09(m，3H)，3.80-3.64(m，3H)，3.32(s，3H)，2.16(d，J＝24.7Hz，6H).

Example 72:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (1- (cyclobutylmethyl) -3, 5-dimethyl 1H-pyrazol-4-yl) -2, 5-difluorophenyl) urea

The preparation method is the same as that of example 54.

LC-MS：503.2，t_R＝2.35min；

¹H NMR(400MHz，MeOD)δ7.97(s，1H)，7.93(dd，J＝11.9，7.0Hz，1H)，7.66(d，J＝7.6Hz，1H)，7.61(d，J＝7.7Hz，1H)，7.46(t，J＝7.5Hz，1H)，7.41-7.34(m，1H)，7.19(s，1H)，6.98(dd，J＝11.4，6.7Hz，1H)，5.52(t，J＝4.8Hz，1H)，4.16(dd，J＝14.4，4.1Hz，1H)，4.07(d，J＝7.2Hz，2H)，3.70(dd，J＝14.4，5.7Hz，1H)，2.82(dt，J＝14.9，7.5Hz，1H)，2.18(s，3H)，2.12(s，3H)，2.10-2.00(m，2H)，1.91(ddt，J＝21.7，17.7，7.5Hz，4H)；

¹⁹F NMR(400MHz，MeOD)δ-119.45，-138.60.

Example 73:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-4- (1- (cyclobutylmethyl) -3, 5- dimethyl-1H-pyrazol-4-yl) -2-fluorophenyl) urea

The preparation method is the same as example 64.

LC-MS：519.1，t_R＝2.46min；

¹H NMR(400MHz，MeOD)δ8.23(d，J＝7.6Hz，1H)，7.97(s，1H)，7.66(d，J＝7.6Hz，1H)，7.61(d，J＝7.6Hz，1H)，7.46(t，J＝7.5Hz，1H)，7.38(t，J＝7.5Hz，1H)，7.19(s，1H)，7.00(d，J＝11.5Hz，1H)，5.52(t，J＝4.8Hz，1H)，4.17(dt，J＝14.4，4.1Hz，1H)，4.07(d，J＝7.1Hz，2H)，3.71(dd，J＝14.4，5.7Hz，1H)，2.82(dt，J＝15.1，7.4Hz，2H)，2.12(s，3H)，2.10-1.99(m，5H)，1.99-1.80(m，4H)；

¹⁹F NMR(400MHz，MeOD)δ-134.86.

Example 74:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-4- (3, 5-dimethyl-1- (oxacyclo) But-3-yl) -1H-pyrazol-4-yl) -2-fluorophenyl) urea

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-bromo-5-chloro-2-fluorophenyl) urea (55mg, 0.126mmol), 3, 5-dimethyl-1- (oxetan-3-yl) -1H-pyrazole-4-boronic acid pinacol ester (70mg, 0.252mmol), Pd (dppf) Cl₂(46mg, 0.0629mmol) and potassium carbonate (35mg, 0.253mmol) were stirred in 1, 4-dioxane/water (10ml/2ml) and the reaction was microwaved to 110 ℃ for 1 hour under nitrogen protection. Cooling to room temperature, adding water, extracting with dichloromethane, drying, and purifying by column chromatographyDissolving to obtain 1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (5-chloro-4- (3, 5-dimethyl-1- (oxetan-3-yl) -1H-pyrazol-4-yl) -2-fluorophenyl) urea (1.4 mg).

LC-MS：507.1，t_R＝2.08min；

¹H NMR(400MHz，MeOD)δ8.24(d，J＝7.6Hz，1H)，7.99(s，1H)，7.63(dd，J＝20.5，7.6Hz，2H)，7.42(dt，J＝33.7，7.4Hz，2H)，7.20(s，1H)，7.00(d，J＝11.4Hz，1H)，5.64-5.55(m，1H)，5.51(s，1H)，5.36(t，J＝4.7Hz，1H)，5.17(t，J＝6.3Hz，1H)，5.11(t，J＝6.3Hz，1H)，5.03(dd，J＝14.0，6.6Hz，2H)，4.17(d，J＝14.3Hz，1H)，3.71(dd，J＝14.3，5.6Hz，1H)，2.13(s，3H)，2.08(s，3H).

Example 75:

1- (2, 5-difluoro-4- (1, 3, 5-trimethyl-1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a]isoindol-5-yl) methyl) urea

The first step is as follows: 1- (4-bromo-2, 5-difluorophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl Yl) preparation of urea

2- (6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) acetic acid (160mg, 0.71mmol), Diphenylphosphorylazide (DPPA) (390mg, 1.4mmol), triethylamine (1mL), 4-bromo-2, 5-difluoroaniline (220mg) were dissolved in anhydrous toluene (15mL), and heated under reflux for 4 hours under nitrogen. LC-MS showed complete reaction, the reaction was concentrated and the residue was dissolved in dichloromethane (20mL) and washed successively with water (2X 10mL), saturated brine (10 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash column chromatography on silica gel and reverse flash column to give 1- (4-bromo-2, 5-difluorophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (70 mg).

The second step is that: 1- (2, 5-difluoro-4- (1, 3, 5-trimethyl-1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazole) Azolo [5, 1-a ]]Preparation of isoindol-5-yl) methyl) urea

1- (4-bromo-2, 5-difluorophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea (36mg, 0.07mmol), 1, 3, 5-trimethyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (42mg, 0.15mmol), Pd (dppf) Cl₂(5mg) and potassium carbonate (20mg, 0.14mmol) were dissolved in a mixture of dioxane (3mL) and water (0.5mL) and heated under reflux for 2 hours under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (10mL) and washed with water (2X 5 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash silica gel column to give 1- (2, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea (6.0 mg).

LC-MS：467.0，t_R＝2.53min；

¹H NMR(400MHz，MeOD)δ7.99(s，1H)，7.89(dd，J＝11.8，6.9Hz，1H)，7.49(t，J＝5.0Hz，2H)，7.23(s，1H)，7.15-7.06(m，1H)，6.97(dd，J＝11.4，6.6Hz，1H)，5.73(t，J＝4.2Hz，1H)，4.27(dd，J＝14.5，3.7Hz，1H)，3.84-3.78(m，1H)，3.76(s，3H)，2.17(s，3H)，2.11(s，3H).

Example 76:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4- (3, 5-dimethyl-1- (oxetan-3-) 1H-pyrazol-4-yl) -2, 5-difluorophenyl) urea

The preparation method is the same as that of example 54.

LC-MS：491.2，t_R：＝2.04min；

¹H NMR(400MHz，MeOD)δ7.85(s，1H)，7.84-7.79(m，1H)，7.53(t，J＝6.7Hz，1H)，7.49(d，J＝7.4Hz，1H)，7.34(t，J＝7.5Hz，1H)，7.25(t，J＝7.5Hz，1H)，7.06(s，1H)，6.86(dd，J＝11.4，6.7Hz，1H)，5.48(dt，J＝14.2，7.0Hz，1H)，5.40(t，J＝4.7Hz，1H)，5.01(t，J＝6.4Hz，2H)，4.94-4.87(m，2H)，4.05(dd，J＝14.4，4.1Hz，1H)，3.58(dd，J＝14.4，5.7Hz，1H)，2.06(s，3H)，2.02(s，3H)；

¹⁹F NMR(376MHz，MeOD)δ-119.44，-138.56.

Example 77:

1- ((5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) -3- (4-3, 5-dimethyl-1H-pyrazol-4-yl) -propanoic acid 2, 5-difluorophenyl) urea

LC-MS：435.1，t_R＝1.92min；

¹H NMR(400MHz，MeOD)δ7.98(s，1H)，7.93(dd，J＝11.9，7.0Hz，1H)，7.66(d，J＝7.5Hz，1H)，7.61(d，J＝7.6Hz，1H)，7.46(t，J＝7.5Hz，1H)，7.38(dd，J＝10.9，4.2Hz，1H)，7.19(s，1H)，7.00(dd，J＝11.5，6.7Hz，1H)，5.52(t，J＝4.8Hz，1H)，4.16(dd，J＝14.4，4.1Hz，1H)，3.70(dd，J＝14.4，5.7Hz，1H)，2.17(s，6H)；

¹⁹F NMR(376MHz，MeOD)δ-119.51，-138.67.

Example 78:

1-((5Himidazo [5, 1-a ]]Isoindol-5-yl) methyl) -3- (4- (3, 5-dimethyl-1- (oxetan-3-) Ylmethyl) -1H-pyrazol-4-yl) -2, 5-difluorophenyl) urea

LC-MS：505.1，t_R＝2.03min；

¹H NMR(400MHz，MeOD)δ9.23(s，1H)，8.05(m，1H)，7.89(m，1H)，7.77m，2H)，7.64(m，2H)，7.17(m，1H)，6.74(s，1H)，5.89(m，1H)，5.36(m，4H)，4.61(d，J＝9.6Hz，2H)，4.40(m，2H)，2.21(t，J＝7.6Hz，3H)，2.06(m，4H).

Example 79:

1- (2, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Different from each other Indol-5-yl) methyl) urea

1- (4-bromo-2, 5-difluorophenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea (36mg, 0.07mmol), 1-methyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (34mg, 0.15mmol), Pd (dppf) Cl₂(5mg) and potassium carbonate (20mg, 0.14mmol) were dissolved in a mixture of dioxane (3mL) and water (0.5mL) and heated under reflux for 2 hours under nitrogen. LC-MS showed the reaction was complete, the reaction was filtered, the filtrate was concentrated, and the residue was dissolved in dichloromethane (10mL) and washed with water (2X 5 mL). The organic phase was dried, filtered and concentrated. The residue was purified by flash silica gel column to give 1- (2, 5-difluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindol-5-yl) methyl) urea (7.0 mg).

LC-MS：439.1，t_R＝2.03min；

¹H NMR(400MHz，MeOD)δ7.96(d，J＝15.5Hz，2H)，7.85(dd，J＝13.1，6.9Hz，1H)，7.80(s，1H)，7.48(dd，J＝13.2，5.2Hz，2H)，7.35(dd，J＝12.0，6.9Hz，1H)，7.21(s，1H)，7.15-7.01(m，1H)，5.70(t，J＝4.3Hz，1H)，4.25(dd，J＝14.5，3.7Hz，1H)，3.92(s，3H)，3.76(dd，J＝14.5，5.3Hz，1H)；

¹⁹F NMR(376MHz，MeOD)δ-120.02，-120.23，-138.39.

Example 80:

1- (4- (1- (cyclopropylmethyl) -1H-pyrazol-4-yl) -2, 5-difluorophenyl) -3- ((6-fluoro-5H-imidazo [5，1-a]Isoindol-5-yl) methyl) urea

The preparation method is the same as that of example 80.

LC-MS：479.2，t_R＝2.47min；

¹H NMR(400MHz，MeOD)δ8.04(d，J＝1.3Hz，1H)，7.98(s，1H)，7.91-7.78(m，2H)，7.54-7.43(m，2H)，7.38(dd，J＝12.0，6.8Hz，1H)，7.21(s，1H)，7.14-7.02(m，1H)，5.70(t，J＝4.4Hz，1H)，4.25(dd，J＝14.5，3.7Hz，1H)，4.03(d，J＝7.1Hz，2H)，3.76(dd，J＝14.5，5.4Hz，1H)，1.38-1.25(m，1H)，0.64(q，J＝5.8Hz，2H)，0.43(q，J＝4.8Hz，2H).

Example 81:

1- (2, 5-difluoro-4- (1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)]Isoindole-5- Yl) methyl) urea

The preparation method is the same as that of example 80.

LC-MS：425.1，t_R＝1.94min；

¹H NMR(400MHz，MeOD)δ8.05-7.93(m，3H)，7.86(dd，J＝13.1，6.9Hz，1H)，7.55-7.44(m，2H)，7.40(dd，J＝12.0，6.9Hz，1H)，7.22(s，1H)，7.10(ddd，J＝9.5，6.8，2.3Hz，1H)，5.71(t，J＝4.5Hz，1H)，4.25(dd，J＝14.5，3.8Hz，1H)，3.77(dd，J＝14.5，5.4Hz，1H)；

¹⁹F NMR(376MHz，MeOD)δ-120.04，-120.36，-138.39.

Example 82:

1- (2, 5-difluoro-4- (1- (oxetan-3-yl) -1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5，1-a]Isoindol-5-yl) methyl) urea

1- (2, 5-difluoro-4- (1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (15mg) and cesium carbonate (100mg) were dissolved in DMF (1mL), 3-iodooxetane (30mg) was stirred overnight at 50 ℃, the reaction solution was cooled to room temperature, and was directly purified by reverse phase column chromatography to give 1- (2, 5-difluoro-4- (1- (oxetan-3-yl) -1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a ] isoindol-5-yl) methyl) urea (3 mg).

LC-MS：481.0，t_R＝2.130min；

¹H NMR(400MHz，MeOD)δ8.14(s，1H)，8.00(dd，J＝12.7，6.0Hz，2H)，7.56(d，J＝7.6Hz，1H)，7.42(td，J＝16.3，9.4Hz，3H)，7.11(t，J＝8.9Hz，1H)，6.43-6.35(m，1H)，5.65-5.58(m，1H)，5.07(d，J＝6.9Hz，4H)，4.61(s，1H)，3.43(d，J＝2.9Hz，1H)，3.38(d，J＝5.4Hz，1H).

Example 83:

1- (4- (1- (cyclobutylmethyl) -1H-pyrazol-4-yl) -2, 5-difluorophenyl) -3- ((6-fluoro-5H-imidazo [5，1-a]Isoindol-5-yl) methyl) urea

The preparation method is the same as that of example 80.

LC-MS：493.1，t_R＝2.32min；

¹H NMR(400MHz，MeOD)δ7.98(d，J＝15.3Hz，2H)，7.91-7.75(m，2H)，7.48(d，J＝7.3Hz，2H)，7.36(dd，J＝11.8，6.8Hz，1H)，7.23(s，1H)，7.11(d，J＝8.7Hz，1H)，5.71(s，1H)，4.25(dd，J＝14.5，3.2Hz，1H)，4.18(d，J＝7.3Hz，2H)，3.77(dd，J＝14.4，5.0Hz，1H)，2.85(dt，J＝14.9，7.5Hz，1H)，2.17-1.74(m，6H).

Example 84:

1- (2, 5-difluoro-4- (1-isobutyl-1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a)] Isoindol-5-yl) methyl) urea

The preparation method is the same as that of example 80.

LC-MS：481.1，t_R＝2.393min；

¹H NMR(400MHz，MeOD)δ7.88(d，J＝14.0Hz，2H)，7.80-7.69(m，2H)，7.37(t，J＝5.1Hz，2H)，7.27(dd，J＝12.0，6.9Hz，1H)，7.12(s，1H)，6.99(dd，J＝11.3，7.5Hz，1H)，5.61(s，1H)，4.15(dd，J＝14.5，3.7Hz，1H)，3.87(d，J＝7.3Hz，2H)，3.67(dd，J＝14.5，5.3Hz，1H)，2.10(dt，J＝13.4，6.8Hz，1H)，0.82(d，J＝6.7Hz，6H).

Example 85:

1- (2, 5-difluoro-4- (2-methoxyethyl) -1H-pyrazol-4-yl) phenyl) -3- ((6-fluoro-5H-imidazo [5, 1-a]isoindol-5-yl) methyl) urea

The preparation method is the same as example 81.

LC-MS：483.1，t_R＝2.08min；

¹H NMR(400MHz，MeOD)δ8.05-7.94(m，2H)，7.86(dd，J＝13.8，6.2Hz，2H)，7.55-7.43(m，2H)，7.38(dd，J＝12.0，6.8Hz，1H)，7.22(s，1H)，7.16-7.02(m，1H)，5.72(t，J＝4.5Hz，1H)，4.34(t，J＝5.2Hz，2H)，4.26(dd，J＝14.4，3.8Hz，1H)，3.87-3.70(m，3H)，3.34(s，3H)；

¹⁹F NMR(400MHz，MeOD)δ-120.07，-120.28，-138.44.

Biological evaluation

One, enzymatic test for IDO activity inhibition

Human indoleamine 2, 3-dioxygenase (IDO) is purchased from BPS Bioscience Inc. Human indoleamine (idoamine) 2, 3-dioxygenase (IDO) enzymatic reaction is carried out in a 96-well plate, the reaction volume is 20 mu L, and the reaction conditions are as follows: 40nM IDO enzyme, 0.2mM L-tryptophan, 50mM KPB (pH 6.5) buffer, 20mM L-sodium ascorbate, 10. mu.M methylene blue, 0.2mg/mL catalase, various concentrations of compound, < 1% dimethyl sulfoxide. After 60 minutes of reaction at 30 ℃, 5 μ L of 30% (W/V) trichloroacetic acid (formulated in 50mM KPB buffer) was added per well and incubated at 50 ℃ for 30 minutes to hydrolyze N-formyl-kynurenine to kynurenine. mu.L of 2% (W/V) p- (dimethylamino) benzaldehyde (p-DMBA)/glacial acetic acid solution was added to each well and absorbance at 490nm was read using a BioTek Synergy H1 microplate reader (Molecular Devices).

A stock solution of the test compound was prepared at 10mM in DMSO, diluted to the highest concentration in the assay in DMSO and then diluted 1: 3 in a gradient, typically 8 to 10 concentration points, each concentration point being provided with a duplicate well, each assay containing 1 reference compound. Analysis enzyme label reader reads 490nm light absorption value original dataCalculating the inhibition of different concentration points of the tested compound on the IDO enzyme activity, and performing nonlinear fitting analysis on the inhibition percentage data by adopting GraphPad Prism software to obtain the half inhibition concentration IC of the compound₅₀The value is obtained.

The activity of the compounds of the examples of the invention was determined by the above test method, enzymological IDO inhibitory activity IC₅₀The results are shown in the following table:

test data show that the compound of the embodiment of the invention has stronger IDO inhibition effect.

Cell model test for inhibition of di, IDO activity

Interferon gamma induces Hela cells to express IDO, and this model was used to test compounds for inhibitory activity against indoleamine 2, 3-dioxygenase (IDO) activity. The culture broth of HeLa cells (ATCC) was phenol red-free RPMI-1640 containing 100. mu.L-tryptophan. The stock solution of the compound to be tested is prepared to be 10mM by using dimethyl sulfoxide, the dimethyl sulfoxide is used for diluting to the highest concentration in the test, the culture medium is used for carrying out 3-fold serial dilution in the test, the dilution is generally carried out to be 8 to 10 concentration points, and each concentration point is provided with a plurality of holes. DMSO final concentration was 0.5% and each experiment contained an internal control compound.

The procedure for the test was: 20,000 Hela cells (ATCC) per well were added to a 96-well plate for overnight culture, and interferon-. gamma. (final concentration of 50ng/mL) and various concentrations of the test compound and the internal reference compound were added to the cultured cells 24 hours later. After 24 hours, 140. mu.L of supernatant/well was transferred to a new 96-well plate, 10. mu.L of 6.1N trichloroacetic acid was added to each well, and incubation was carried out at 50 ℃ for 30 minutes to hydrolyze N-formyl-kynurenine to kynurenine. The reaction mixture was centrifuged (2500 rpm for 10 min) to remove the precipitate, 100. mu.L of the supernatant was transferred to another new 96-well plate, 100. mu.L of a 2% (W/V) p- (dimethylamino) benzaldehyde (p-DMBA)/glacial acetic acid solution was added to each well, and the 490nm absorbance was read using a BioTek Synergy H1 microplate reader (Molecular Devices).

Reading 490nm light absorption value original data by an analytical microplate reader, calculating inhibition of different concentration points of a tested compound on the activity of the IDO enzyme, and performing nonlinear fitting analysis on inhibition percentage data by adopting GraphPad Prism software to obtain half inhibition concentration IC of the compound₅₀The value is obtained.

The activity of the compounds of the examples of the invention was determined by the above test method, cytological IDO inhibitory activity IC₅₀The results are shown in the following table:

test data show that the compound of the embodiment of the invention has stronger cell activity.

Claims

1. A compound of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

wherein,

R₁selected from hydrogen, deuterium, halogen;

R₂selected from hydrogen, deuterium, halogen or pyrazolyl, wherein said pyrazolyl is further substituted by one or more groups selected from halogen, cyano, optionally substituted or unsubstituted C_1-3Alkyl is substituted by a substituent;

R、R₃、R₄each independently selected from hydrogen, deuterium, halogen or C_1-3An alkyl group;

the optionally substituted substituents are selected from deuterium, halogen, hydroxy、C_1-3Alkyl, cyclopropyl, cyclobutyl, pyrrolidinyl, morpholinyl, C_1-3Alkoxy or phenyl;

m is 0 or 1;

n is 2 or 3;

and excluding the following compounds:

2. the compound of formula (I), its stereoisomers, or a pharmaceutically acceptable salt thereof, according to claim 1, wherein R is₁Selected from hydrogen atoms.

3. A compound of formula (I), its stereoisomers, or a pharmaceutically acceptable salt thereof, according to claim 1, wherein R, R₃、R₄Each independently selected from hydrogen atoms.

4. A compound of formula (I), its stereoisomers, or a pharmaceutically acceptable salt thereof, according to claim 1, wherein R is₂Selected from hydrogen, halogen or pyrazolyl; wherein said pyrazolyl is further substituted by C_1-3Alkyl groups are substituted.

5. A compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof as claimed in claim 1, selected from compounds of formula (ii):

wherein:

R、R₁、R₂and n is as defined in claim 1.

6. The following compounds, stereoisomers thereof or pharmaceutically acceptable salts thereof:

7. a pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 6, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

8. Use of a compound of any one of claims 1 to 6, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 7, for the manufacture of a medicament for the treatment of a disease characterized by a pathology of a tryptophan metabolism disorder mediated by IDO/TDO; the disease characterized by the pathology of a disorder of IDO/TDO-mediated tryptophan metabolism is selected from the group consisting of cancer or tumor, viral infection, depression, neurodegenerative disorders, trauma, age-related cataract, organ transplant rejection or autoimmune disease.