Aromatic fused ring compound, pharmaceutical composition containing aromatic fused ring compound and application of aromatic fused ring compound

Abstract

The invention provides an aromatic fused ring compound, a pharmaceutical composition containing the compound and application of the aromatic fused ring compound, and relates to the technical field of medicines. The aromatic fused ring compound disclosed by the invention has a structure as shown in a formula (I): # imgabs0 # or a stereoisomer and pharmaceutically acceptable salt thereof, wherein the structural unit # imgabs1 # is selected from # imgabs2 # or # imgabs3 #, and each substituent group is defined in the specification. The aromatic fused ring compound has a strong inhibition effect on SIKs enzyme activity, has a positive regulation effect on TNF-alpha and IL-10 in serum of mice with acute inflammation, has a longer half-life period and a higher oral exposure amount, has a remarkable inhibition effect on NF-kappa B gene expression of THP-1 cells under MDP stimulation, and can be used for preparing medicines for treating acute inflammation. The traditional Chinese medicine composition has a remarkable weight recovery promoting effect on cachexia induced by cis-platinum.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to aromatic fused ring compounds, pharmaceutical compositions containing the compounds and applications thereof. Technical Background

[0002] Salt-inducible kinase (SIK) belongs to the AMP-activated protein kinase (AMPK) family and includes three isoforms, namely SIK1, SIK2 and SIK3. The activity of SIK can be affected by many different forms of post-translational modifications, including LKB1, CaMK, PKA-mediated phosphorylation, p300-mediated acetylation, etc. The SIK family regulates multiple aspects of physiology, including metabolism, neurophysiology, immune function and development. Recent studies have shown that SIK activity is at the center of the kinase cascade that regulates sleep and circadian rhythms, which in turn are master regulators of all physiology and behavior, including metabolism, immune function and neurophysiology. Based on the very diverse biological functions of SIKs, the potential application space of SIKs targeted drugs may include tumors, inflammatory diseases, obesity-induced diabetes and other diseases related to kinase activity disorders: Elevated IL-6 levels are strongly correlated with giant cell arteritis and large-scale arteritis. SIKs inhibitors can inhibit the polarization of macrophages and the secretion of proinflammatory factors such as IL-6, and have the potential to treat giant cell arteritis and large-scale arteritis; SIKs are upregulated in neointimal lesions during arterial restenosis. The use of small molecule SIKs inhibitors can inhibit the migration and proliferation of vascular smooth muscle cells (SMCs), as well as the formation of new endothelium, and have the potential to treat arterial stenosis; SIKs inhibitors can inhibit Toll-like receptor-mediated innate immune activation and have the potential to treat acute respiratory distress syndrome; in studies on vitiligo, SIKs inhibitors can trigger the skin to produce dark pigments, increase the expression of MITF and its target genes, and animal models also show that they have the potential to treat vitiligo.

[0003] At present, inhibitors targeting SIKs targets can be divided into non-selective inhibitors and selective inhibitors. HG-9-91-01, YKL-05-099, and MRT-199665 are all first-generation pan-SIKs inhibitors with poor selectivity for members of each subfamily, and most drugs have defects such as off-target and poor pharmacokinetic properties. In addition, subtype-selective SIK2 / 3 inhibitors represented by ARN-3261 are still in early clinical or preclinical studies. In summary, it is of great significance to further develop new SIK inhibitors and fully study their in vivo mechanisms of action.

[0004] RIPK2 kinase is an emerging drug target for the treatment of autoimmune and inflammatory diseases. RIPK2 is mainly involved in regulating the signal transduction of NOD1 and NOD2 pathways and the activation of their downstream nuclear factor NF-κB and mitogen-activated protein kinase MAPK pathways, thereby regulating a series of inflammatory responses in the body. The inhibitory effect of compounds on RIPK2 kinase can be examined at the cellular level by detecting the downstream gene NF-κB or cytokine TNF-α (Nachbur U., Stafford C.A., Bankovacki A., et al. ARIPK2 inhibitor delays NOD signalling events yet prevents inflammatory cytokine production. Nat Commun. 2015, 6: 6442.). Currently, there are no RIPK2 inhibitors on the market for disease treatment. Compounds such as WEHI-345 and GSK583 have been shown to have good RIPK2 kinase inhibitory activity and inflammatory treatment potential. The development of RIPK2 inhibitors has good application prospects for autoimmune and inflammatory disease drugs. Summary of the invention

[0005] In view of the problems existing in the prior art, the present invention provides an aromatic fused ring compound, a pharmaceutical composition comprising the compound and the application thereof. The compound obtained by the present invention can inhibit the activity of SIKs and can be used for the treatment of related diseases.

[0006] The present invention adopts the following technical solution

[0007] The present invention provides a compound having a structure shown in Formula I, its stereoisomers, and pharmaceutically acceptable salts:

[0008]

[0009] in:

[0010] Structural unit Selected from or

[0011] Y and W are each independently selected from C or N;

[0012] V, Z are selected from CH or N;

[0013] X is selected from N or CR g ;

[0014] W, Y, and Z are not all N; W, Y, and Z are not all C;

[0015] L1 is selected from O or NH or CH2;

[0016] L2 is selected from C 1-3 Alkanes;

[0017] R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, unsubstituted or methyl-substituted 5-6-membered aromatic heterocyclic group, unsubstituted or methyl-substituted 9-10-membered fused heteroaromatic ring group, 4, 5 or 6-membered heterocyclic group, the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, the 4, 5 or 6-membered heterocyclic group can be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution;

[0018] R a , R b Each independently selected from H, C 1-3 Alkyl;

[0019] R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2;

[0020] m is selected from 1, 2 or 3;

[0021] R4 is selected from H, halogen, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy, -NHS(O)2R d 、-NR e S(O)2R d , -S(O)2R d ;

[0022] R d , R e Each independently selected from C 1-3 Alkyl, cyclopropyl;

[0023] R5 is selected from -C(O)R f 、-S(O)2Rf 、-C(O)NHR f ;

[0024] R f Selected from C 1-3 Alkyl, cyclopropyl, halogen-substituted C 1-3 Alkyl, -CH2OH;

[0025] R6, R7, R8, R g Each independently selected from H, F, Cl, methyl, methoxy, trifluoromethoxy, difluoromethoxy, trifluoromethyl; R7 and R8 may form Wherein R6, V, and X are as defined above;

[0026] R 10 and R 11 Each independently selected from H, C 1-3 Alkyl, one or more fluorine-substituted C 1-3 Alkyl, F,

[0027]

[0028] The present invention preferably has a structure shown in Formula II, or a stereoisomer or a pharmaceutically acceptable salt thereof:

[0029]

[0030] in:

[0031] Y and W are each independently selected from C or N;

[0032] Z is selected from CH or N;

[0033] W, Y, and Z are not all N; W, Y, and Z are not all C;

[0034] L1 is selected from O or NH or CH2;

[0035] L2 is selected from C 1-3 Alkanes;

[0036] R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b, -OH, unsubstituted or methyl-substituted 5-6-membered aromatic heterocyclic group, unsubstituted or methyl-substituted 9-10-membered fused heteroaromatic ring group, 4, 5 or 6-membered heterocyclic group, the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, the 4, 5 or 6-membered heterocyclic group can be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution;

[0037] R a , R b Each independently selected from H, C 1-3 Alkyl;

[0038] R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2;

[0039] m is selected from 1, 2 or 3;

[0040] R4 is selected from H, F, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy, -NHS(O)2R d ;

[0041] R5 is selected from -C(O)R f 、-S(O)2R f 、-C(O)NHR f ;

[0042] R f Selected from C 1-3 Alkyl, cyclopropyl, -CF3, -CH2CF3, -CH2OH;

[0043] R d Selected from C 1-3 Alkyl, cyclopropyl.

[0044] The present invention preferably has a structure shown in Formula III, or a stereoisomer or a pharmaceutically acceptable salt thereof:

[0045]

[0046] in:

[0047] Y and W are each independently selected from C or N;

[0048] Z is selected from CH or N;

[0049] V is selected from CH or N;

[0050] X is selected from N or CR g ;

[0051] W, Y, and Z are not all N; W, Y, and Z are not all C;

[0052] L1 is selected from O or NH or CH2;

[0053] L2 is selected from C 1-3 Alkanes;

[0054] R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, unsubstituted or methyl-substituted 5-6-membered aromatic heterocyclic group, unsubstituted or methyl-substituted 9-10-membered fused heteroaromatic ring group, 4, 5 or 6-membered heterocyclic group, the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, the 4, 5 or 6-membered heterocyclic group can be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution;

[0055] R a , R b Each independently selected from H, C 1-3 Alkyl;

[0056] R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2;

[0057] m is selected from 1, 2 or 3;

[0058] R4 is selected from F, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy, -NHS(O)2R d 、-NR e S(O)2Rd 、- -S(O)2R d ;

[0059] R d Selected from C 1-3 Alkyl, cyclopropyl;

[0060] R6, R7, R8, R g Each independently selected from H, F, Cl, methyl, methoxy, trifluoromethoxy, difluoromethoxy, trifluoromethyl; R7 and R8 may form wherein R6, V, and X are as defined above.

[0061] The present invention preferably has a structure shown in Formula IV, or a stereoisomer or a pharmaceutically acceptable salt thereof:

[0062]

[0063] in:

[0064] Y and W are each independently selected from C or N;

[0065] Z is selected from CH or N;

[0066] W, Y, and Z are not all N; W, Y, and Z are not all C;

[0067] L1 is selected from O or NH or CH2;

[0068] L2 is selected from C 1-3 Alkanes;

[0069] R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, 4, 5 or 6-membered heterocyclic group, wherein the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, and the 4, 5 or 6-membered heterocyclic group may be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution;

[0070] R a , R b Each independently selected from H, C 1-3 Alkyl;

[0071] R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2;

[0072] m is selected from 1, 2 or 3;

[0073] R4 is selected from F, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy;

[0074] R 10 and R 11 Each independently selected from H, C 1-3 Alkyl, one or more fluorine-substituted C 1-3 Alkyl, F,

[0075]

[0076] The present invention preferably has the structure shown in the following formula V or formula VI or formula VII or formula VIII or formula IX, or a stereoisomer or a pharmaceutically acceptable salt thereof:

[0077]

[0078] in:

[0079] L1 is selected from O or NH or CH2;

[0080] L2 is selected from C 1-3 Alkanes;

[0081] R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, 4, 5 or 6-membered heterocyclic group, wherein the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, and the 4, 5 or 6-membered heterocyclic group may be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C1-3 Alkyl, C 1-3 Alkoxy substitution;

[0082] R a , R b Each independently selected from H, C 1-3 Alkyl;

[0083] R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2;

[0084] m is selected from 1, 2 or 3;

[0085] R 10 and R 11 Each independently selected from H, C 1-3 Alkyl, one or more fluorine-substituted C 1-3 Alkyl, F,

[0086]

[0087] Preferably, The unit is selected from the following structures:

[0088]

[0089] Preferably, The unit is selected from the following structures:

[0090]

[0091] R6, R7, R8, R g Each independently selected from H, F, Cl, methyl, methoxy, trifluoromethoxy, difluoromethoxy, trifluoromethyl; R7 and R8 may form

[0092] More preferably, the compound is selected from:

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099] More preferably, the compound is selected from:

[0100]

[0101] The second aspect of the present invention provides a pharmaceutical composition comprising the above-mentioned compound, its isomers, or pharmaceutically acceptable salts thereof, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, adjuvant or solvent, and the pharmaceutical composition is in the form of tablets, capsules, pills, granules, powders, suppositories, injections, solutions, suspensions, ointments, patches, lotions, drops, liniments or sprays.

[0102] The third aspect of the present invention is to provide the use of the above-mentioned compound, its isomers, and pharmaceutically acceptable salts thereof in the preparation of drugs for treating or preventing diseases caused by abnormal activation of SIK kinase, and preventing or treating diseases accompanied by weight loss.

[0103] Preferably, the disease caused by abnormal activation of SIK kinase is selected from at least one of cancer, immune inflammation, diabetes, and central nervous system diseases. The cancer is selected from at least one of ovarian cancer, breast cancer, prostate cancer, glioma, and diffuse large B-cell lymphoma; the immune inflammation is selected from at least one of arthritis, colitis, psoriasis, atopic dermatitis, psoriatic arthritis, systemic lupus erythematosus, osteoarthritis, complications caused by organ transplantation, autoimmune thyroid disease, Sjögren's syndrome, vitiligo, autoimmune kidney damage, autoimmune liver damage, chronic obstructive pulmonary disease, multiple sclerosis, muscular lateral sclerosis, giant cell arteritis, Takayasu arteritis, and sepsis; the central nervous system disease is selected from at least one of depression, insomnia, and epilepsy.

[0104] Preferably, the disease associated with weight loss is selected from at least one of cachexia, growth retardation, wasting, and skeletal muscle dystrophy. The cachexia is selected from at least one of cachexia associated with cancer, acquired immunodeficiency syndrome, heart failure, and / or chronic kidney disease.

[0105] Compared with the prior art, the present invention has the following beneficial technical effects: (1) The compounds of the present invention have excellent in vitro inhibitory activity against kinases SIK1 / 2 / 3, and the effects of some compounds are significantly better than those of comparative compounds; (2) The representative compounds of the present invention can reduce TNFα and increase IL-10 in LPS-induced acute inflammation in mice, and show certain anti-inflammatory activity in animals; (3) The compounds of the present invention have good oral pharmacokinetic properties, and compared with comparative compound 2, have a longer half-life and a higher oral exposure; (4) The representative compounds of the present invention have significantly better inhibitory activity against RIPK2 kinase, inhibitory activity against TNF-α secretion in BMDM cells, and inhibitory activity against NF-κB gene expression in THP-1 cells than positive drugs WEHI-345 and GLPG3970; (5) The representative compounds of the present invention have a significant weight-increasing effect on ICR young mice, and have a promoting function on the growth of leg bone length in mice; and have a significant weight-recovery effect on cisplatin-induced cachexia.

[0106] Terms and Definitions:

[0107] Structural unit Here, N refers to the nitrogen atom. The structural unit must comply with the valence rules of carbon atoms or nitrogen atoms and belong to a 6-membered and 5-membered aromatic condensed ring. W, Y, and Z are not all carbon atoms, nor are they all nitrogen atoms.

[0108] The term "isomer" means that the compounds of the present invention may exist in specific geometric or stereoisomer forms. In the present invention, isomers mainly refer to the (R)- and (S)-enantiomers and racemates that appear in the connection between R1 and L2 in R1-L2, and the (R)- and (S)-enantiomers and racemates that appear in the substituents in the R1 group. The isomers produced in these two places and their mixtures are all included in the scope of the present invention.

[0109] Unless otherwise specified, C 1-3 Alkanes represent straight or branched saturated hydrocarbon groups, which may be monosubstituted (e.g., -CH2CH3, -CH2F) or polysubstituted (e.g., -CH(CH2), -CF3,), and may be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). Examples of alkyl groups include methyl (Me), ethyl (Et), and propyl (e.g., n-propyl and isopropyl).

[0110] Unless otherwise specified, a numerical range represents all integers including the numbers at both ends of the range. Unless otherwise specified, an integer from 0 to 3 represents 0, 1, 2, 3; an integer from 7 to 12 represents 7, 8, 9, 10, 11, 12; 1 to 2 represents 1, 2; 5 to 6 represents 5, 6, and so on. 1-3 Examples of alkanes include, but are not limited to, methyl, ethyl, propyl, isopropyl.

[0111] (CH2)n (n is an integer selected from 0, 1, 2, 3) may be substituted by 1-2 methyl groups, and examples of R1-L2 include but are not limited to:

[0112]

[0113] (wherein R1 is other substituents as defined herein).

[0114] 5-6 membered aromatic heterocyclic rings and 9-10 membered fused heteroaromatic rings contain carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O and S, the total number of S and O atoms in the aromatic heterocyclic ring does not exceed 1, examples include but are not limited to: benzimidazolyl, benzopyrazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, chromanyl, benzophenone ... linyl, furanyl, imidazolinyl, imidazolyl, pyrazolyl, pyrrolyl, thienyl, thiazolyl, thiadiazolyl, oxazolyl, 1H-indazolyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyrimidinyl, pyridinyl, pyridazinyl, pyrazinyl, phenoxazinyl, phthalazinyl.

[0115] 4, 5 or 6 membered heterocyclic radicals, wherein the heteroatom is nitrogen, oxygen, at least one nitrogen atom, the nitrogen and sulfur heteroatoms may be optionally oxidized (NO and S(O)p, p is 1 or 2). The nitrogen atom may be substituted or unsubstituted (i.e. N or NR, wherein R is H or other substituents as defined herein). The heterocyclic ring may be attached to any side group of a heteroatom or carbon atom to form a stable structure. Examples include, but are not limited to, morpholinyl, piperazinyl, tetrahydropyrrolyl, piperidinyl, piperidonyl, 4-piperidonyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, pyrrolidinyl, 2H-pyrrolyl, pyrrolyl.

[0116] Examples of 7-12 membered spirocycloalkane groups include, but are not limited to:

[0117]

[0118] Examples of 7-12 membered spirocyclic heteroalkanes include, but are not limited to:

[0119]

[0120] Examples of 7-12 membered bridged cycloalkane groups include, but are not limited to:

[0121]

[0122] Examples of 7-12 membered bridged heterocycloalkane groups include, but are not limited to:

[0123] Examples of 8-12 membered cycloalkane groups include, but are not limited to:

[0124] Examples of 8-12 membered cycloheteroalkyl groups include, but are not limited to:

[0125] BRIEF DESCRIPTION OF THE DRAWINGS

[0126] Figure 1 :The inhibitory activity of the compound on TNF-α secretion from BMDM cells stimulated by MDP;

[0127] Figure 2 : The inhibitory activity of the compounds on NF-κB gene expression in THP-1 cells stimulated by MDP;

[0128] Figure 3 :Effects of compounds on body weight and leg bone length in ICR mice;

[0129] Figure 4 : Effects of compounds on body weight in mice with cisplatin-induced cachexia. DETAILED DESCRIPTION

[0130] The feasibility of the present invention is illustrated below by using embodiments. Those skilled in the art should understand that according to the teachings of the prior art, modifications or replacements of corresponding technical features still fall within the scope of protection claimed by the present invention.

[0131] Example 1: Preparation of 8-methoxy-6-(7-((piperidin-4-methylene)amino)imidazo[1,2-a]pyridin-3-yl)-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A1)

[0132]

[0133] Step 1: Synthesis of intermediate 1-2

[0134] 2-Amino-4-bromopyridine (2.275 g) and sodium bicarbonate (2.18 g) were placed in a round-bottom flask, followed by the addition of 24 mL of anhydrous ethanol to fully dissolve the raw materials, followed by the addition of 5.3 mL of chloroacetaldehyde solution (40% wt aqueous solution), and stirred and refluxed at 82°C overnight. After the reaction was completed, the reaction solution was filtered through a sand core funnel / diatomaceous earth and extracted three times with ethyl acetate / water. After the solvent was removed in vacuo, the product was purified by column chromatography (PE:EA=1:2) to obtain 1.27 g (1-2) of a brown solid. ESI-MS: m / z=198 [M+H] + .

[0135] Step 2: Synthesis of intermediate 1-3

[0136] Add intermediate 1-2 (394 mg), 1-tert-butyloxycarbonyl-4-aminomethylpiperidine (129 mg), 36 mg Pd2(dba)3, 69 mg Xantphos, 288 mg t-BuONa, and 3 mL Toluene to a two-necked bottle, seal, replace N2, and react overnight at 100°C. After the reaction is completed, quench the reaction with EA. Add 50 mL of water and extract three times, and the organic phase is dried over anhydrous Na2SO4. The organic phase is spin-dried and purified by column chromatography (PE:EA=1:2~1:3) to obtain the product as a brown-yellow solid (1-3). ESI-MS: m / z=331[M+H] + .

[0137] Step 3: Synthesis of intermediate 1-4

[0138] The intermediate 1-3 (2 mmol) was dissolved in 5 mL DCM, and (Boc)2O (2.4 mmol), 4-dimethylaminopyridine (0.4 mmol), and triethylamine (3 mmol) were added under ice bath. After the addition, the mixture was stirred under ice bath for 10 min and then transferred to room temperature for reaction for 6 h. After the reaction was completed, the organic phases were combined after extraction with EA three times, and the solvent was removed by vacuum distillation. The mixture was separated by column chromatography (PE:EA) = 4:1 to obtain a light yellow solid (1-4). ESI-MS: m / z = 431 [M+H] + .

[0139] Step 4: Synthesis of intermediate 1-5

[0140] The intermediate 1-4 (2 mmol) was dissolved in 4 mL DMF, and NIS (2.4 mmol) was added under ice bath. After the reaction was completed, the reaction was quenched with Na2S2O3, extracted with EA three times, and the organic phases were combined. The solvent was removed by vacuum distillation, and column chromatography (PE:EA) = 4:1 was used to obtain a brown solid (1-5). ESI-MS: m / z = 557 [M+H] + .

[0141] Step 5: Synthesis of Intermediate 1-7

[0142] Dissolve the compound 5-bromo-7-fluoro-1-indanone (1-6, 3g, 13.1mmol) in 25mL DCM, add methanesulfonic acid (22.7g, 15.5mL) under ice bath conditions, and then slowly add NaN3 (1.7g, 26.2mmol) in batches. After reacting for 2h under ice bath, add 20% NaOH (58mL) and continue to react for 1 hour under ice bath. After the reaction is completed, extract with DCM, combine the organic phases, dry with anhydrous magnesium sulfate, and remove the solvent by vacuum distillation to obtain a solid, which is directly used in the next step.

[0143] Step 6: Synthesis of Intermediate 1-8

[0144] To a stirred solution of intermediate 1-7 (3 g, 12.29 mmol) in THF (30 mL) was added dropwise a 25 w% MeONa solution in MeOH (3.35 mL, 14.75 mmol). The reaction mixture was stirred at room temperature for 2 hours, quenched with saturated NH4Cl aqueous solution and THF was removed by distillation under reduced pressure. The solid obtained in the remaining aqueous phase was filtered to obtain the desired compound, which was used directly in the next step.

[0145] Step 7: Synthesis of Intermediate 1-9

[0146] LiHMDS (1M THF solution, 15 mmol) was added dropwise to a THF solution of intermediate 1-9 (2.56 g, 10 mmol) at 0°C, and the mixture was stirred at 0°C for 30 min. Subsequently, a triflate reagent (15 mmol) was added, and the reaction mixture was heated to room temperature and reacted for 20 h. LiHMDS and triflate reagents were added until the conversion no longer increased. The reactants were then quenched with water or saturated NH4Cl aqueous solution, THF was concentrated, and the aqueous phase was extracted with EtOAc. The combined organic phases were dried over a desiccant and filtered, and purified by column chromatography (PE:EA=1:1) to give a light yellow compound (1-9). ESI-MS: m / z=338[M+H] + .

[0147] Step 8: Synthesis of Intermediate 1-10

[0148] The intermediate 1-9 (156 mg, 0.46 mmol) was dissolved in DMSO solution (2 mL), and diboric acid pinacol ester (142 mg, 0.56 mmol) and potassium acetate (147 mg, 1.5 mmol) were added, and the mixture was reacted at 80°C overnight under nitrogen protection. After the reaction, the organic phases were combined by EA extraction three times, dried over anhydrous sodium sulfate, and then distilled under reduced pressure to remove the solvent, and purified by column chromatography (PE:EA=2:1) ​​to obtain compound 1-10. ESI-MS: m / z=386[M+H] + .

[0149] Step 9: Preparation of 8-methoxy-6-(7-((piperidin-4-methylene)amino)imidazo[1,2-a]pyridin-3-yl)-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A1)

[0150] Intermediate 1-5 (167 mg, 0.3 mmol) and intermediate 1-10 (116 mg, 0.3 mmol) were added to 1,4-dioxane solvent (3 mL), and (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (Pd(dppf)Cl2) (22 mg, 0.03 mmol) and cesium carbonate (326 mg, 1 mmol) were added, and the mixture was refluxed under nitrogen protection overnight. After the reaction, the solvent was removed by distillation under reduced pressure, and the mixture was purified by column chromatography (DCM:MeOH=30:1). After obtaining the intermediate, the protecting group was removed by trifluoroacetic acid under ice bath conditions, and the mixture was purified by column chromatography (DCM:MeOH=15:1) to obtain compound A1. 1 H NMR (400MHz, DMSO-d6) δ8.38(d,J=7.5Hz,1H),7.57(s,1H),7.05(dd,J=15.1,1.6Hz,2H),6.60 (t,J=5.9Hz,1H),6.53(dd,J=7.5,2.2Hz,1H),6.35(d,J=2.3Hz,1H),4.29(q,J=9.6Hz,2H),3.8 3(s,3H),3.56(d,J=6.2Hz,2H),3.24(d,J=12.4Hz,2H),3.00(t,J=6.0Hz,2H),2.92(t,J=6.1Hz ,2H),2.81(t,J=12.4Hz,2H),1.88(d,J=12.6Hz,3H),1.39-1.25(m,2H).ESI-MS:m / z=488[M+H] + .

[0151] Example 2: Preparation of 6-(6-(((1s,4s)-4-aminocyclohexyl)amino)imidazo[1,2-a]pyridin-3-yl)-8-methoxy-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A2)

[0152]

[0153] Synthesis step 1: Synthesis of intermediate 2-2

[0154] Synthesis steps: Refer to step 1 of Example 1, and use 2-amino-5-bromopyridine instead of 2-amino-4-bromopyridine. Yield: 85%; ESI-MS: m / z=198[M+1]+.

[0155] Synthesis step 2: Synthesis of intermediate 2-3

[0156] Synthesis steps: Referring to step 2 of Example 1, 1-N-Boc-cis-1,4-cyclohexanediamine was used instead of 1-tert-butyloxycarbonyl-4-aminomethylpiperidine to obtain intermediate 2-3. Yield: 45%; ESI-MS: m / z=331[M+1]+.

[0157] Synthesis step 3: Synthesis of intermediate 2-4

[0158] The synthesis steps refer to step 4 of Example 1, and intermediate 2-3 is used instead of 1-4 to obtain intermediate 2-4, which is stored in the dark and used immediately after preparation. Yield: 23%; ESI-MS: m / z=457[M+1]+.

[0159] Synthesis step 4: Preparation of 6-(6-(((1s,4s)-4-aminocyclohexyl)amino)imidazo[1,2-a]pyridin-3-yl)-8-methoxy-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A2)

[0160] Synthesis steps: Refer to step 9 of Example 1, use intermediate 2-4 instead of intermediate 1-5, yield: 20%; 1H NMR (400MHz, Methanol-d4) δ7.70 (d, J = 2.1Hz, 1H), 7.61 (s, 1H), 7.40 (dd, J = 9.6 ,0.7Hz,1H),7.23(d,J=1.5Hz,1H),7.14-7.03(m,2H),4.32(q,J=9.3Hz,2H),3.9 1(s,3H),3.69(t,J=6.2Hz,2H),3.36(s,1H),3.04(t,J=6.2Hz,2H),2.90(dq,J=8 .5,4.3,3.8Hz,1H),1.91-1.80(m,2H),1.76-1.52(m,7H).ESI-MS:m / z=488[M+1] + .

[0161] Example 3: Preparation of 6-(6-(((1s,4s)-4-aminocyclohexyl)amino))pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A3)

[0162]

[0163] Synthesis step 1: Synthesis of intermediate 3-2

[0164] Synthesis steps: Refer to step 2 of Example 1, replace 1-tert-butyloxycarbonyl-4-aminomethylpiperidine with 1-N-Boc-cis-1,4-cyclohexanediamine, and replace intermediate 1-2 with 6-bromopyrazolo[1,5-A]pyridine to obtain intermediate 3-2. Yield: 42%; ESI-MS: m / z=331[M+1] + .

[0165] Synthesis step 2: Synthesis of intermediate 3-3

[0166] Synthesis steps: Refer to step 4 of Example 1, replace 1-4 with intermediate 3-2 to obtain intermediate 3-3, store in the dark, and use it as it is prepared. Yield: 23%; ESI-MS: m / z=457[M+1] + .

[0167] Synthesis step 3: Preparation of 6-(6-(((1s,4s)-4-aminocyclohexyl)amino))pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A3)

[0168] Synthesis steps: Refer to step 9 of Example 1, use intermediate 3-3 instead of intermediate 1-5, yield: 20%; ESI-MS: m / z=488[M+1]+ .

[0169] Example 4: Preparation of 8-methoxy-6-(5-((piperidin-4-methylene)amino)pyrazolo[1,5-a]pyridin-3-yl)-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A4)

[0170]

[0171] Synthesis step 1: Synthesis of intermediate 4-2

[0172] Synthesis steps: Refer to step 2 of Example 1, and replace intermediate 1-2 with 5-bromopyrazolo[1,5-A]pyridine to obtain intermediate 4-2. Yield: 42%; ESI-MS: m / z=331[M+1] + .

[0173] Synthesis step 2: Synthesis of intermediate 4-3

[0174] Synthesis steps: Referring to step 3 of Example 1, intermediate 4-2 was used instead of intermediate 1-3 to obtain intermediate 4-3, yield: 84%; ESI-MS: m / z=431[M+1] + .

[0175] Synthesis step 3: Synthesis of intermediate 4-4

[0176] Synthesis steps: Refer to step 4 of Example 1, replace intermediate 1-4 with intermediate 4-3 to obtain intermediate 4-4, which was stored in the dark and used immediately after preparation. Yield: 73%; ESI-MS: m / z=557[M+1] + .

[0177] Synthesis Step 4: Preparation of 8-methoxy-6-(5-((piperidin-4-methylene)amino)pyrazolo[1,5-a]pyridin-3-yl)-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1(2H)-one (A4)

[0178] Synthesis steps: Refer to step 9 of Example 1, use intermediate 4-4 instead of intermediate 1-5, yield: 30%; ESI-MS: m / z = 488 [M+1] + .

[0179] Example 5: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyrimidin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A5)

[0180]

[0181] Synthesis step 1: Synthesis of intermediate 5-2

[0182] 6-Bromopyrazolo[1,5-A]pyrimidine (200 mg, 1 mmol) was dissolved in 5 mL of methanol, KOH (336 mg, 6 mmol) was added, and the mixture was heated to 65°C. After the reaction was completed, the mixture was moved to room temperature, and the solvent was removed by distillation under reduced pressure. After purification by column chromatography (PE:EA=2:1), intermediate 5-2 was obtained with a yield of 82%; ESI-MS: m / z=136[M+1] + .

[0183] Synthesis step 2: Synthesis of intermediate 5-3

[0184] The intermediate 5-2 (2 mmol) was dissolved in DMF solvent (5 mL), 4-(2-bromoethyl)morpholine hydrobromide (3 mmol) and cesium carbonate (6 mmol) were added, and the mixture was heated to 80°C and reacted overnight. The solvent was removed by distillation under reduced pressure, and the intermediate 5-3 was obtained after purification by column chromatography (DCM:MeOH=20:1) with a yield of 51%; ESI-MS: m / z=249[M+1] + .

[0185] Synthesis step 3: Synthesis of intermediate 5-4

[0186] Synthesis steps: Refer to step 4 of Example 1, replace intermediate 1-4 with intermediate 5-3 to obtain intermediate 5-4, which was stored in the dark and used immediately after preparation. Yield: 63%; ESI-MS: m / z=375[M+1] + .

[0187] Synthesis step 4: Synthesis of intermediate 5-5

[0188] Intermediate 1-8 (1020 mg, 4 mmol) was dissolved in THF (10 mL). Under nitrogen protection, 10 mL of BH3·Me2S (2N in THF) was added in an ice bath. After the mixture was brought to room temperature, the temperature was gradually raised to 78°C and the reaction was allowed to proceed overnight. After the reaction was completed, the mixture was placed in an ice bath, 1N HCl solution was added to quench the reaction, and the pH was adjusted to neutral. After three extractions with EA, the organic phases were combined, the solvent was removed by distillation under reduced pressure, and intermediate 5-5 was obtained after purification by column chromatography. ESI-MS: m / z=242[M+1] + .

[0189] Synthesis step 5: Synthesis of intermediate 5-6

[0190] Dissolve intermediate 5-5 (1.2 g, 5 mmol), pyridine (10 mmol), and DMAP (0.25 mmol) in DCM, add trifluoroacetic anhydride (2.1 g, 6 mmol) under ice bath, and react overnight at room temperature. After the reaction, remove part of the solvent and reagent by vacuum distillation, and adjust the pH to neutral with saturated sodium bicarbonate solution. After EA extraction three times, combine the organic phases, remove the solvent by vacuum distillation, and purify by column chromatography to obtain intermediate 5-6. ESI-MS: m / z=338[M+1] + .

[0191] Synthesis Step 6: Synthesis of Intermediate 5-7

[0192] Synthesis steps: Referring to step 8 of Example 1, intermediate 5-6 was used to replace intermediate 1-9 to obtain intermediate 5-7, yield: 63%; ESI-MS: m / z=386[M+1] + .

[0193] Synthesis step 7: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyrimidin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A5)

[0194] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 5-4, and replace intermediate 1-10 with intermediate 5-7. Yield: 30%; 1 H NMR(400MHz,Chloroform-d)δ8.47(s,1H),8.30(s,1H),8.27(d,J=2.7Hz,1H),7.85(d,J=10.9Hz,2H),7.21(q,J=8.3,7.7Hz,1H),4. 80(d,J=17.4Hz,2H),4.15(s,2H),3.90(d,J=17.5Hz,2H),3.76(s,4H),3.05(s,2H),2.88(s,2H),2.60(s,4H).ESI-MS:m / z=506[M+1] + .

[0195] Example 6: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A6)

[0196]

[0197] Synthesis step 1: Synthesis of intermediate 6-1

[0198] Synthesis steps: Referring to step 8 of Example 1, intermediate 3-1 was used instead of intermediate 1-9 to obtain intermediate 6-1, yield: 63%; ESI-MS: m / z=245[M+1] + .

[0199] Synthesis step 2: Synthesis of intermediate 6-2

[0200] Intermediate 6-1 (732 mg, 3 mmol) was dissolved in THF (10 mL), and H2O2 (1.02 g, 9 mmol) and NaOH (360 mg, 4.5 mL, 9 mmol) were added under ice bath conditions. The reaction was completed after 30 min. The solvent was removed by distillation under reduced pressure and the mixture was directly used for the next step.

[0201] Synthesis step 3: Synthesis of intermediate 6-3

[0202] Synthesis steps: Referring to step 2 of Example 5, intermediate 6-2 was used to replace intermediate 5-2 to obtain intermediate 6-3, yield: 42%; ESI-MS: m / z=248[M+1] + .

[0203] Synthesis step 4: Synthesis of intermediate 6-4

[0204] Synthesis steps: Refer to step 4 of Example 1, replace intermediate 1-4 with intermediate 6-3 to obtain intermediate 6-4, which was stored in the dark and used immediately after preparation. Yield: 56%; ESI-MS: m / z=374[M+1] + .

[0205] Synthesis Step 5: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A6)

[0206] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 6-4, and replace intermediate 1-10 with intermediate 5-7, yield: 30%; 1 H NMR(400MHz,Chloroform-d)δ8.10(s,1H),8.02(s,1H),7.66(d,J=9.9Hz,1H),7.09-6.99(m,1H),6.94(s,1H),6.91(s,1H), 4.75(s,2H),4.13(s,2H),3.88(d,J=26.1Hz,5H),3.76(s,4H),2.98(s,2H),2.86(s,2H),2.60(s,4H).ESI-MS:m / z=505[M+1] + .

[0207] Example 7: Preparation of 2,2,2-trifluoro-1-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A7)

[0208]

[0209] Synthesis step 1: Synthesis of intermediate 7-2

[0210] Synthesis steps: Referring to step 4 of Example 5, 6-bromo-1,2,3,4-tetrahydroisoquinoline was used to replace intermediate 5-5 to obtain intermediate 7-2, yield: 63%; ESI-MS: m / z=245[M+1] + .

[0211] Synthesis step 2: Synthesis of intermediate 7-3

[0212] Synthesis steps: Referring to step 8 of Example 1, intermediate 7-2 was used instead of intermediate 1-9 to obtain intermediate 7-3, yield: 63%; ESI-MS: m / z=356[M+1] + .

[0213] Synthesis step 3: Preparation of 2,2,2-trifluoro-1-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A7)

[0214] Synthesis steps: Referring to step 9 of Example 1, intermediate 6-4 was used to replace intermediate 1-5, and intermediate 7-3 was used to replace intermediate 1-10 to obtain compound A7, yield: 30%; 1 H NMR(400MHz,Chloroform-d)δ8.10(s,1H),8.02(s,1H),7.67(d,J=9.7Hz,1H),7.44(s,1H),7.35(s,1H),7.20(d,J=7.9Hz,1H),7.04(d,J=9.7H z,1H),4.81(d,J=18.2Hz,2H),4.13(s,2H),3.91(d,J=16.6Hz,2H),3.76(s,4H),3.03(s,2H),2.86(s,2H),2.61(s,4H).ESI-MS: m / z=475[M+1] + .

[0215] Example 8: Preparation of 2,2,2-trifluoro-1-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyrimidin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A8)

[0216]

[0217] Synthesis step 1: Preparation of 2,2,2-trifluoro-1-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyrimidin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A8)

[0218] Synthesis steps: Referring to step 9 of Example 1, intermediate 5-4 was used to replace intermediate 1-5, and intermediate 7-3 was used to replace intermediate 1-10 to obtain compound A8, yield: 30%; 1 H NMR(400MHz,Chloroform-d)δ8.47(s,1H),8.30(s,1H),8.27(d,J=2.7Hz,1H),7.85(d,J=10.9Hz,2H),7.21(q,J=8.3,7.7Hz,1H),4. 80(d,J=17.4Hz,2H),4.15(s,2H),3.90(d,J=17.5Hz,2H),3.76(s,4H),3.05(s,2H),2.88(s,2H),2.60(s,4H).ESI-MS:m / z=476[M+1] + .

[0219] Example 9: Preparation of 1-(8-methoxy-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyrimidin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A9)

[0220]

[0221] Synthesis step 1: Synthesis of intermediate 9-1

[0222] Synthesis steps: Referring to step 4 of Example 5, acetyl chloride was used instead of trifluoroacetic anhydride to obtain intermediate 9-1, yield: 63%; ESI-MS: m / z=284[M+1] + .

[0223] Synthesis step 2: Synthesis of intermediate 9-2

[0224] Synthesis steps: Referring to step 8 of Example 1, intermediate 9-1 was used to replace intermediate 1-9 to obtain intermediate 9-2, yield: 63%; ESI-MS: m / z=332[M+1] + .

[0225] Synthesis step 3: Preparation of 1-(8-methoxy-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyrimidin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A9)

[0226] Synthesis steps: Referring to step 9 of Example 1, intermediate 5-4 was used to replace intermediate 1-5, and intermediate 9-2 was used to replace intermediate 1-10 to obtain compound A9, yield: 30%; 1 H NMR(400MHz,Chloroform-d)δ8.46(s,1H),8.29(s,1H),8.26(d,J=2.8Hz,1H),7.44(d,J=13.6Hz,1H),7.37(s,1H),4.62(d,J=50.5Hz ,2H),4.15(s,2H),3.93(d,J=12.5Hz,3H),3.88-3.65(m,6H),2.91(d,J=28.8Hz,4H),2.60(s,4H),2.22(s,3H).ESI-MS:m / z=452[M+1] + .

[0227] Example 10: 2,2,2-Trifluoro-1-(8-methoxy-6-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A10)

[0228]

[0229] Synthesis step 1: Synthesis of intermediate 10-2

[0230] Dissolve tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate (10-1, 1.15 g, 5 mmol) in DCM (10 mL), add triethylamine TEA (1.4 mL, 10 mmol) and methanesulfonyl chloride (0.6 mL, 7.5 mmol) under ice bath conditions, react at room temperature for 6 h, wash with water, extract with EA, and distill under reduced pressure to remove the solvent. The obtained liquid is directly used in the next reaction.

[0231] Synthesis step 2: Synthesis of intermediate 10-3

[0232] Synthesis steps: Referring to step 2 of Example 5, intermediate 6-2 was used to replace intermediate 5-2, and intermediate 10-2 was used to replace 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 10-3, yield: 63%; ESI-MS: m / z=347[M+1] + .

[0233] Synthesis step 3: Synthesis of intermediate 10-4

[0234] The intermediate 10-3 (2 mmol) was dissolved in DCM (4 mL), and 2 mL of trifluoroacetic acid was added under ice bath conditions, and the reaction was carried out at room temperature for 4 to 6 hours. After the reaction was completed, the solvent was removed by distillation under reduced pressure, and the intermediate was directly used for the next reaction.

[0235] Synthesis Step 4: Synthesis of Intermediate 10-5

[0236] The intermediate 10-4 (2 mmol) was dissolved in dichloromethane, and formaldehyde (150 mg, 5 mmol) and 1 mL of glacial acetic acid were added. After reacting at 40°C for 1 h, NaBH(OAc)3 (1232 mg, 6 mmol) was added. The reaction was stopped overnight, and the pH was adjusted to acidic with hydrochloric acid solution and then extracted. The aqueous phase was adjusted to alkaline with saturated NaHCO3 solution and then extracted with EA. The organic phases were combined and the solvent was removed under reduced pressure. The product was purified by silica gel column chromatography with DCM:MeOH (25:1) as the eluent to obtain a light yellow solid (10-5) with a yield of 67%; ESI-MS: m / z=261[M+H]+.

[0237] Synthesis Step 5: Synthesis of Intermediate 10-6

[0238] Synthesis steps: Refer to step 4 of Example 1, replace intermediate 1-4 with intermediate 10-5 to obtain intermediate 10-6, which was stored in the dark and used immediately after preparation. Yield: 56%; ESI-MS: m / z=387[M+1] + .

[0239] Synthesis step 6: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A10)

[0240] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 10-6, and replace intermediate 1-10 with intermediate 5-7. Yield: 30%; ESI-MS: m / z=518[M+1] + .

[0241] Example 11: 2,2,2-trifluoro-1-(8-methoxy-6-(6-(piperidin-4-ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A11)

[0242]

[0243] Synthesis step 1: Synthesis of intermediate 11-1

[0244] Synthesis steps: Referring to step 2 of Example 5, intermediate 6-2 was used instead of intermediate 5-2, and 1-BOC-4-bromomethylpiperidine was used instead of 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 11-1, yield: 53%; ESI-MS: m / z=332[M+1] + .

[0245] Synthesis step 2: Synthesis of intermediate 11-2

[0246] Synthesis steps: Refer to step 4 of Example 1, replace intermediate 1-4 with intermediate 11-1 to obtain intermediate 11-2, which is stored in the dark and used as is. Yield: 56%; ESI-MS: m / z=458[M+1] + .

[0247] Synthesis step 3: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(6-(piperidin-4-ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A11)

[0248] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 11-2, and replace intermediate 1-10 with intermediate 5-7. Yield: 30%; ESI-MS: m / z=489[M+1] + .

[0249] Example 12: 2,2,2-Trifluoro-1-(8-methoxy-6-(5-(piperidin-4-ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A12)

[0250]

[0251] Synthesis step 1: Synthesis of intermediate 12-3

[0252] 4-Methoxypyridine (12-1, 10.9 g, 10 mmol) and 2,4-dinitrophenylhydroxy (12-2, 20.9 g, 11.5 mmol) were added to DCM (30 mL), stirred at room temperature, and after the reaction was completed as monitored by TLC, the solvent was removed by distillation under reduced pressure and used directly in the next step.

[0253] Synthesis step 2: Synthesis of intermediate 12-4

[0254] Intermediate 12-3 and methyl propiolate (10 g, 12 mmol) were dissolved in DMF (30 mL), potassium carbonate (16.6 g, 12 mmol) was added, and the mixture was reacted at 80°C. After the reaction was completed, water was added and the mixture was extracted three times with EA. The organic phases were combined and the solvent was removed by vacuum distillation. The mixture was purified by column chromatography (PE:EA=1:1) to obtain intermediate 12-4. ESI-MS: m / z=207[M+1] + .

[0255] Synthesis Step 3: Synthesis of Intermediate 12-5

[0256] Add 125 mL of 50% H2SO4 aqueous solution to the intermediate 12-4 (14 g), stir at 120°C for 3-6 h, adjust the pH to neutral with NaOH after the reaction is completed, extract with ether 3-5 times, combine the organic phases, remove the solvent by vacuum distillation, and purify by column chromatography (DCM: MeOH = 20: 1) to obtain the intermediate 12-5, ESI-MS: m / z = 135 [M+1] + .

[0257] Synthesis Step 4: Synthesis of Intermediate 12-6

[0258] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used to replace intermediate 5-2, and 1-BOC-4-bromomethylpiperidine was used to replace 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 12-6, yield: 53%; ESI-MS: m / z=332[M+1] + .

[0259] Synthesis Step 5: Synthesis of Intermediate 12-7

[0260] The synthesis steps refer to step 4 of Example 1 to obtain intermediate 12-7.

[0261] Synthesis Step 6: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(5-(piperidin-4-ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A12)

[0262] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 12-7, and replace intermediate 1-10 with intermediate 5-7, yield: 30%; 1H NMR(400MHz,Chloroform-d)δ8.32(s,1H),8.01(s,1H),6.90(d,J=13.0Hz,3H),6.50(s,1H),4.75(s,2H),3.88(d,J=23.9Hz,7H),3.32(d ,J=12.3Hz,2H),2.99(s,2H),2.80(t,J=12.6Hz,2H),2.05(s,1H),1.95(d,J=13.6Hz,2H),1.55(d,J=13.7Hz,2H).ESI-MS: m / z=489[M+1] + .

[0263] Example 13: 2,2,2-Trifluoro-1-(8-methoxy-6-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A13)

[0264]

[0265] Synthesis step 1: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(6-(piperidin-4-ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A13)

[0266] Synthesis steps: Referring to Example 12, 4-(2-bromoethyl)morpholine hydrobromide was used instead of 1-BOC-4-bromomethylpiperidine to obtain compound A13, yield: 30%; 1 H NMR(400MHz, DMSO-d6)δ8.58(dd,J=7.6,0.5Hz,1H),8.26(d,J=1.0Hz,1H),7.42-7.35(m, 4H),7.26(dd,J=8.6,2.5Hz,2H),7.18(dd,J=4.3,2.6Hz,1H),7.10-7.06(m,2H),6.64(dd ,J=7.6,2.6Hz,1H),4.65(d,J=18.4Hz,2H),4.26-4.21(m,3H),3.86-3.80(m,2H),3.58(t ,J=4.7Hz,5H),2.96(dt,J=13.0,5.8Hz,2H),2.73(t,J=5.7Hz,2H).ESI-MS:m / z=505[M+1] + .

[0267] Example 14: 2,2,2-Trifluoro-1-(8-methoxy-6-(5-((piperidin-4-ethoxy)amino)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A14)

[0268]

[0269] Synthesis step 1: Synthesis of intermediate 14-2

[0270] Pyrazolo[1,5-A]pyridine-5-amine (532 mg, 4 mmol), 1-Boc-4-piperidinic acid (917 mg, 4 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 920 mg, 4.8 mmol), 1-hydroxybenzotriazole (HOBT, 649 mg, 4.8 mmol) and N,N-diisopropylethylamine (DIPEA, 775 mg, 6.0 mmol) were added to DCM (12 mL) and reacted at room temperature overnight. After the reaction was completed, the solvent was distilled off under reduced pressure, water was added, and EA was used for extraction three times. The organic phases were combined and purified by column chromatography (PE:EA=1:1) to obtain intermediate 14-2, ESI-MS: m / z=345[M+1] + .

[0271] Synthesis step 2: Synthesis of intermediate 14-3

[0272] To a stirred solution of intermediate 14-2 (2 mmol) in THF (10 mL) was added BH3.Me2S (2.1 mL) at 0°C. The mixture was heated to 60°C and stirred at this temperature for 2 hours. The reaction progress was monitored by TLC. After the starting material was completely consumed, the reaction mass was cooled to 0°C, quenched with methanol, and then stirred again at 60°C for 2 hours. The organic phases were combined after extraction with EA three times and purified by column chromatography to obtain intermediate 14-3, ESI-MS: m / z=291[M+1] + .

[0273] Synthesis step 3: Synthesis of intermediate 14-4

[0274] Synthesis steps: Refer to step 4 of Example 1, replace 1-4 with intermediate 14-3 to obtain intermediate 14-4, which was stored in the dark and used immediately after preparation. Yield: 36%; ESI-MS: m / z=457[M+1] + .

[0275] Synthesis step 4: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(5-((piperidin-4-ethoxy)amino)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A14)

[0276] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 14-4, and replace intermediate 1-10 with intermediate 5-7. Yield: 30%; ESI-MS: m / z=488[M+1] + .

[0277] Example 15: 2,2,2-trifluoro-1-(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A15)

[0278]

[0279] Synthesis step 1: Synthesis of intermediate 15-2

[0280] Synthesis procedure: Referring to step 1 of Example 10, 4-Boc-2-hydroxymethylmorpholine was used instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 15-2, which was directly used in the next step.

[0281] Synthesis step 2: Synthesis of intermediate 15-3

[0282] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used to replace 5-2, and intermediate 15-2 was used to replace 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 15-3, yield: 63%; ESI-MS: m / z=334[M+1] + .

[0283] Synthesis Step 3: Synthesis of Intermediate 15-4

[0284] Synthesis steps: Refer to step 4 of Example 1, replace intermediate 1-4 with intermediate 15-3 to obtain intermediate 15-4, which was stored in the dark and used immediately after preparation. Yield: 47%; ESI-MS: m / z=460[M+1] + .

[0285] Synthesis step 4: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A15)

[0286] The synthesis steps refer to step 9 of Example 1, intermediate 15-4 is used instead of intermediate 1-5, and intermediate 5-7 is used instead of intermediate 1-10. Yield: 30%; ESI-MS: m / z=491[M+1]+.

[0287] Example 16: 2,2,2-trifluoro-1-(6-(5-(2-(4-(2-hydroxyethyl)piperazin-1-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A16)

[0288]

[0289] Synthesis step 1: Synthesis of intermediate 16-1

[0290] Synthesis procedure: Refer to step 1 of Example 10, and use 10-2 instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 16-1, which is directly used in the next step.

[0291] Synthesis step 2: Synthesis of intermediate 16-2

[0292] Synthesis steps: Refer to step 3 of Example 10, use intermediate 16-1 instead of 10-3 to obtain intermediate 16-2, which is directly used in the next step.

[0293] Synthesis step 3: Synthesis of intermediate 16-3

[0294] Intermediate 16-2 (180 mg, 0.73 mmol) was dissolved in acetonitrile, and 2-bromoethanol (183 mg, 1.46 mmol) and potassium carbonate (309 mg, 2.22 mmol) were added and reacted at 90° C. After the reaction was completed, the solvent was removed by distillation under reduced pressure and the product was directly used in the next step.

[0295] Synthesis Step 4: Synthesis of Intermediate 16-4

[0296] Synthesis steps: Refer to step 4 of Example 1, replace 1-4 with intermediate 16-3 to obtain intermediate 16-4, which was stored in the dark and used immediately after preparation. Yield: 47%; ESI-MS: m / z=417[M+1] + .

[0297] Synthesis Step 5: Preparation of 2,2,2-trifluoro-1-(6-(5-(2-(4-(2-hydroxyethyl)piperazin-1-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A16)

[0298] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 16-4, and replace intermediate 1-10 with intermediate 5-7, yield: 30%; 1 H NMR(400MHz,Chloroform-d)δ8.32(s,1H),8.03(d,J=13.7Hz,1H),7.50(d,J=37.1Hz,1H),6.93(s,1H),6.88(s,1H),6.53(s,1H),4.74(d,J=10.1Hz,1 H),4.14(s,2H),3.89(d,J=20.7Hz,4H),3.64(s,2H),2.99(s,2H),2.87(s, 2H),2.62(d,J=20.7Hz,8H),2.07(d,J=45.8Hz,4H).ESI-MS:m / z=548[M+1] + .

[0299] Example 17: (S)-2,2,2-trifluoro-1-(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A17)

[0300]

[0301] Synthesis steps Referring to Example 15, 4-Boc-2-hydroxymethylmorpholine was replaced with N-tert-butyloxycarbonyl-(S)-2-morpholinemethanol; 1 H NMR (400MHz, Methanol-d4) δ8.39(d,J=7.6Hz,1H),8.11(d,J=2.7Hz,1H),7.13( t,J=2.7Hz,1H),7.04-6.98(m,1H),6.65(dd,J=7.6,2.6Hz,1H),4.72(d,J=16.0H z,1H),4.10(dd,J=4.9,1.9Hz,1H),3.98-3.86(m,4H),3.02-2.95(m,1H),2.93- 2.80(m,2H),1.37-1.27(m,2H),1.32(s,3H),1.28(s,5H).ESI-MS:m / z=491[M+1] + .

[0302] Example 18: (R)-2,2,2-trifluoro-1-(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A18)

[0303]

[0304] Synthesis steps Referring to Example 15, N-tert-butyloxycarbonyl-(R)-2-morpholinemethanol was used instead of 4-Boc-2-hydroxymethylmorpholine; 1 H NMR(400MHz,Chloroform-d)δ8.31(d,J=7.6Hz,1H),8.00(d,J=1.1Hz,1H),6.9 9-6.84(m,2H),6.54(ddd,J=7.6,2.7,1.3Hz,1H),4.73(d,J=9.8Hz,2H),4.07- 3.95(m,1H),3.98-3.91(m,0H),3.93(s,2H),3.95-3.88(m,3H),3.85(t,J=4.9 Hz,1H),3.70(td,J=11.2,3.0Hz,1H),3.09-2.72(m,5H).ESI-MS:m / z=491[M+1] + .

[0305] Example 19: Preparation of 2-hydroxy-1-(8-methoxy-6-(7-((piperidin-4-methylene)amino)imidazo[1,2-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A19)

[0306]

[0307] Synthesis step 1: Synthesis of intermediate 19-1

[0308] Dissolve intermediate 5-5 (1.0 equiv., 242 mg) in 7 mL DCM, add glycolic acid (1.2 equiv.), HOBt (1.2 equiv.), and EDCI (1.2 equiv.) in turn under ice bath, and stir for 5 min. Dissolve N-methylmorpholine in 2 mL DCM and add dropwise to the reaction solution. After the addition is completed, return the reaction to room temperature and react overnight. After the reaction is completed, spin dry the solvent and separate by column chromatography. ESI-MS: m / z = 300 [M+H] + .

[0309] Synthesis Step 2: Synthesis of Intermediate 19-2

[0310] Synthesis steps: Referring to step 8 of Example 1, intermediate 19-1 was used to replace intermediate 1-9 to obtain intermediate 19-2, yield: 63%; ESI-MS: m / z=348[M+1] + .

[0311] Synthesis step 3: Preparation of 2-hydroxy-1-(8-methoxy-6-(7-((piperidin-4-methylene)amino)imidazo[1,2-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A19)

[0312] Synthesis steps: Referring to step 9 of Example 1, intermediate 19-2 was used instead of 1-10 to obtain compound A19, yield: 30%; ESI-MS: m / z=450[M+1] + .

[0313] Example 20: Preparation of (S)-2-hydroxy-1-(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A20)

[0314]

[0315] Synthesis steps: Referring to step 9 of Example 1, intermediate 17-4 was used to replace 1-5, and intermediate 19-2 was used to replace 1-10 to obtain compound A20, yield: 30%; ESI-MS: m / z=453[M+1] + .

[0316] Example 21: Cyclopropyl(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)methanone (A21)

[0317]

[0318] Synthesis step 1: Synthesis of intermediate 21-1

[0319] Synthesis steps: Refer to step 1 of Example 19, use cyclopropionic acid instead of glycolic acid to obtain intermediate 21-1, which is directly used in the next step.

[0320] Synthesis step 2: Synthesis of intermediate 21-2

[0321] Synthesis steps: Referring to step 8 of Example 1, intermediate 21-1 was used to replace intermediate 1-9 to obtain intermediate 21-2, yield: 63%; ESI-MS: m / z=358[M+1] + .

[0322] Synthesis step 3: Preparation of cyclopropyl (8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)methanone (A21)

[0323] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 15-4, and replace intermediate 1-10 with intermediate 21-2. Yield: 30%; ESI-MS: m / z=463[M+1] + .

[0324] Example 22: 2-(((3-(2-(cyclopropanesulfonyl)-8-methoxy-1,2,3,4-tetrahydroisoquinolin-6-yl)pyrazolo[1,5-a]pyridin-5-yl)oxy)methyl)morpholine (A22)

[0325]

[0326] Synthesis Procedure Referring to Example 21, cyclopropanesulfonyl chloride was used instead of cyclopropionic acid to obtain Compound A22. 1 HNMR(400MHz,Methanol-d4)δ8.35(d,J=7.6Hz,1H),8.07(s,1H),7.09(d,J=2.7Hz,1H),6.94(s,2H) ,6.62(dd,J=7.6,2.6Hz,1H),4.37(s,2H),4.10(dd,J=4.7,1.4Hz,2H),3.97(dddt,J=9.5,7.4,4.8,2 .1Hz,2H),3.89(s,3H),3.73(ddd,J=12.1,10.1,4.1Hz,1H),3.54(t,J=5.8Hz,2H),2.96(dt,J=11.4, 4.6Hz,4H),2.51(tt,J=7.9,4.9Hz,1H),1.11-1.06(m,2H),1.02-0.96(m,2H).ESI-MS:m / z=499[M+1] + .

[0327] Example 23: 2,2,2-trifluoro-1-(8-methoxy-6-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A23)

[0328]

[0329] Synthesis step 1: Synthesis of intermediate 23-2

[0330] Synthesis steps: Referring to step 1 of Example 1, 2-amino-4-fluoropyridine was used instead of 2-amino-4-bromopyridine to obtain intermediate 23-2, ESI-MS: m / z=137 [M+H] + .

[0331] Synthesis Step 2: Synthesis of Intermediate 23-3

[0332] Synthesis steps: Referring to step 2 of Example 5, intermediate 23-2 was used to replace intermediate 5-2 to obtain intermediate 23-3, ESI-MS: m / z=334 [M+H] + .

[0333] Synthesis Step 3: Synthesis of Intermediate 23-4

[0334] Synthesis steps: Refer to step 4 of Example 1, replace 1-4 with intermediate 23-3 to obtain intermediate 23-4, store in the dark, and use it as it is prepared. Yield: 23%; ESI-MS: m / z=460[M+1] + .

[0335] Synthesis step 4: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A23)

[0336] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 23-4, and replace intermediate 1-10 with intermediate 5-7. Yield: 30%; ESI-MS: m / z=491[M+1] + .

[0337] Example 24: 1-(6-(5-(2-amino-2-methylpropoxy)pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethanol-1-one (A24)

[0338]

[0339] Synthesis steps: Refer to Example 15, using N-BOC-2-amino-2-methyl-1-propanol instead of 4-Boc-2-hydroxymethylmorpholine; ESI-MS: m / z=463[M+1] + .

[0340] Example 25: (S)-2,2,2-trifluoro-1-(8-methoxy-6-(5-(pyrrole-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A25)

[0341]

[0342] Synthesis steps: Refer to Example 15, using N-BOC-2-amino-2-methyl-1-propanol instead of 4-Boc-2-hydroxymethylmorpholine; ESI-MS: m / z=475[M+1]+ .

[0343] Example 26: 1-(6-(5-(((1s,4s)-4-aminocyclohexane)oxy)pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethanol-1-one (A26)

[0344]

[0345] Synthesis steps: Refer to Example 15. The configuration of intermediates 26-2 and 26-3 will be reversed, and trans-N-BOC-4-aminocyclohexanol will be used instead of 4-Boc-2-hydroxymethylmorpholine; ESI-MS: m / z = 489 [M+1] + .

[0346] Example 27: 2,2,2-trifluoro-1-(8-methoxy-6-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A27)

[0347]

[0348] Synthesis steps: Refer to Example 15, using 1-methyl-4-hydroxymethylpyrazole instead of 4-Boc-2-hydroxymethylmorpholine; ESI-MS: m / z=486[M+1] + .

[0349] Example 28: 1-(6-(5-(2-(dimethylamino)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethyl-1-one (A28)

[0350]

[0351] Synthesis steps: Refer to Example 15, using dimethylaminoethyl chloride hydrochloride instead of 4-Boc-2-hydroxymethylmorpholine; ESI-MS: m / z=463[M+1] + .

[0352] Example 29: 1-(6-(5-((1H-indazol-7-yl)methoxy)pyrazolo[1,5-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethyl-1-one (A29)

[0353]

[0354] Synthesis steps: Refer to Example 15, using 1H-indazole-7-methanol instead of 4-Boc-2-hydroxymethylmorpholine; ESI-MS: m / z=522[M+1] + .

[0355] Example 30: Preparation of 2,2,2-trifluoro-1-(8-methoxy-6-(7-((piperidin-4-methylene)amino)imidazo[1,2-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one (A30)

[0356]

[0357] Synthesis steps: Refer to step 9 of Example 1, and use intermediate 5-7 instead of 1-10 to obtain compound A30. Yield: 30%; ESI-MS: m / z=488[M+1] + .

[0358] Example 31: (S)-8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-N-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinoline 2(1H)-carboxamide (A31)

[0359]

[0360] Synthesis step 1: Synthesis of intermediate 31-1

[0361] 2,2,2-trifluoroethylamine (112 mg, 1.13 mmol) was added to a mixture of CDI (172 mg, 1.06 mmol) in tetrahydrofuran (4 mL) cooled to 0°C. After 1 hour, HPLC analysis showed that CDI had been consumed. In a separate container, a mixture of intermediate 5-5 (169 mg, 0.708 mmol) and K2HPO4 (123 mg, 0.708 mmol) in 2-MeTHF (2 mL) and water (2 mL) was stirred at 25°C, and the pH of the aqueous layer was adjusted to 9.0, and the water contained 20% KOH. The above reaction solution was added to the liquid containing intermediate 5-5 and reacted at room temperature for 2 hours. After the reaction was completed, 5% citric acid was added and then extracted three times with EA. The organic phases were combined and the solvent was removed by distillation under reduced pressure. After column chromatography purification (PE:EA=2:1), intermediate 31-1 was obtained. ESI-MS: m / z=367[M+1] + .

[0362] Synthesis step 2: Synthesis of intermediate 31-2

[0363] Synthesis steps: Referring to step 8 of Example 1, intermediate 31-1 was used to replace intermediate 1-9 to obtain intermediate 31-2, ESI-MS: m / z=415[M+1] + .

[0364] Synthesis step 3: Preparation of (S)-8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-N-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinoline 2(1H)-carboxamide (A31)

[0365] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 17-4, and replace intermediate 1-10 with intermediate 31-2. Yield: 32%; ESI-MS: m / z=520[M+1] + .

[0366] Example 32: (S)-3,3,3-trifluoro-1-(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)propan-1-one (A32)

[0367]

[0368] Synthesis step 1: Synthesis of intermediate 32-1

[0369] Synthesis steps: Refer to step 1 of Example 19, use 3,3,3-trifluoropropionic acid instead of glycolic acid to obtain intermediate 32-1, which is directly used in the next step.

[0370] Synthesis step 2: Synthesis of intermediate 32-2

[0371] Synthesis steps: Referring to step 8 of Example 1, intermediate 32-1 was used instead of 1-9 to obtain intermediate 32-2, yield: 63%; ESI-MS: m / z=400[M+1] + .

[0372] Synthesis step 3: Preparation of (S)-3,3,3-trifluoro-1-(8-methoxy-6-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-3,4-dihydroisoquinolin-2(1H)-yl)propan-1-one (A32)

[0373] The synthesis steps refer to step 9 of Example 1, using intermediate 17-4 instead of intermediate 1-5, and intermediate 32-2 instead of intermediate 1-10, yield: 38%; ESI-MS: m / z=505[M+1]+.

[0374] Example 33: 1-(6-(7-((2-(diethylamino)ethyl)amino)imidazo[1,2-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethanol-1-one (A33)

[0375]

[0376] Synthesis step 1: Synthesis of intermediate 33-1

[0377] Synthesis steps: Referring to step 2 of Example 1, 1-tert-butyloxycarbonyl-4-aminomethylpiperidine was replaced with N,N-diethylethylenediamine to obtain intermediate 33-2, ESI-MS: m / z=233 [M+H] + .

[0378] Synthesis step 2: Synthesis of intermediate 33-2

[0379] Synthesis steps: Referring to step 3 of Example 1, intermediate 33-1 was used to replace intermediate 1-3 to obtain intermediate 33-2, ESI-MS: m / z=333 [M+H] + .

[0380] Synthesis Step 3: Synthesis of Intermediate 33-3

[0381] Synthesis steps: Refer to step 4 of Example 1, replace intermediate 1-4 with intermediate 33-2 to obtain intermediate 33-3, store in dark, and use it as it is prepared. Yield: 43%; ESI-MS: m / z=460[M+1] + .

[0382] Synthesis step 4: Preparation of 1-(6-(7-((2-(diethylamino)ethyl)amino)imidazo[1,2-a]pyridin-3-yl)-8-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethanol-1-one (A33)

[0383] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 33-3, and replace intermediate 1-10 with intermediate 5-7. Yield: 30%; ESI-MS: m / z=490[M+1] + .

[0384] Example 34: N-(2',4'-difluoro-5-(7-((piperidin-4-methylene)amino)imidazo[1,2-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A34)

[0385]

[0386] Synthesis step 1: Synthesis of intermediate 34-2

[0387] Synthesis steps: Referring to step 9 of Example 1, intermediate 34-1 was used to replace intermediate 1-5, and 2,4-difluorophenylboronic acid was used to replace intermediate 1-10 to obtain intermediate 34-2, ESI-MS: m / z=284 [M+H] + .

[0388] Synthesis Step 2: Synthesis of Intermediate 34-3

[0389] Intermediate 34-2 (2.16 g) was dissolved in 8 mL of pyridine and 8 mL of dichloromethane, followed by the addition of cyclopropanesulfonyl chloride (1.29 g), and stirred at 40°C for 4 hours. The pH was adjusted to a weak base with dilute hydrochloric acid, and then extracted three times with ethyl acetate / water. The solvent was removed by distillation under reduced pressure, and column chromatography (PE:EA) = 20:3 was used to obtain 1.5 g of a white solid. ESI-MS: m / z = 388 [M+H] + .

[0390] Synthesis step 3: Preparation of N-(2',4'-difluoro-5-(7-((piperidin-4-methylene)amino)imidazo[1,2-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A34)

[0391] Intermediate 1-4 (430 mg, 1 mmol) and intermediate 34-3 (387 mg, 1 mmol) were dissolved in DMF (4 mL), and Pd(dppf)Cl2 (36 mg, 0.05 mmol) and KOAc (196 mg, 2 mmol) were added. The mixture was protected by nitrogen and reacted at 110°C for 12 h. After the reaction, the mixture was washed with water and extracted with EA three times. The organic phases were combined and the solvent was removed by vacuum distillation. The intermediate was purified by column chromatography (DCM: MeOH = 30:1). The protective group was removed by hydrochloric acid / 1,4-dioxane solution (4 M) at 25°C. The reaction was terminated after 1 h. The compound A34 was obtained by column chromatography purification (DCM: MeOH = 15:1). ESI-MS: m / z = 538 [M+H] + .

[0392] Example 35: N-(2',4'-difluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A35)

[0393]

[0394] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used instead of intermediate 1-4 to obtain compound A35, 1HNMR (400MHz, Methanol-d4) δ8.39(d,J=7.6Hz,1H),8.15(s,1H),7.64(t,J=1.8Hz,1H),7.56(td,J=8.9,6.5Hz,1H),7. 47(q,J=1.6Hz,1H),7.24(q,J=2.0Hz,2H),7.13-7.00(m,2H),6.65(dd,J=7.6,2.6Hz,1H),4.14-4.02(m,2H),3.90(ddd, J=13.9,7.8,3.5Hz,2H),3.65(ddd,J=11.7,9.5,4.7Hz,1H),3.02(dd,J=12.5,2.4Hz,1H),2.83(dd,J=9.7,2.9Hz,2H), 2.74(dd,J=12.5,10.6Hz,1H),2.63(tt,J=7.9,4.8Hz,1H),1.10-1.03(m,2H),1.00-0.94(m,2H).ESI-MS:m / z=541[M+H] + .

[0395] Example 36: N-(2',4'-difluoro-5-(5-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A36)

[0396]

[0397] Synthesis steps Referring to step 3 of Example 34, intermediate 13-1 was used instead of intermediate 1-4 to obtain compound A36, 1 HNMR(400MHz,Chloroform-d)δ8.28(dd,J=7.6,0.6Hz,1H),8.08(s,1H),7.58(t,J=1.8Hz,1 H),7.51-7.38(m,2H),7.21(q,J=1.7Hz,1H),7.16(d,J=2.6Hz,1H),7.02-6.89(m,2H),6.49 (dd,J=7.6,2.6Hz,1H),4.28(t,J=5.7Hz,2H),3.79(t,J=4.7Hz,4H),2.93(t,J=5.7Hz,2H), 2.73(s,4H),2.61-2.55(m,1H),1.01-0.94(m,2H),0.87-0.84(m,2H).ESI-MS:m / z=555[M+H] + .

[0398] Example 37: N-(2'-Fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-4'-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A37)

[0399]

[0400] Synthesis Step 1: Synthesis of Intermediate 37-2

[0401] Synthesis steps: Refer to step 8 of Example 1, replace intermediate 1-9 with intermediate 34-1 to obtain intermediate 37-2.

[0402] Synthesis Step 2: Synthesis of Intermediate 37-3

[0403] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with 37-2, and replace intermediate 1-10 with 4-bromo-3-fluorotrifluorotoluene to obtain intermediate 37-3. ESI-MS: m / z=334 [M+H] + .

[0404] Synthesis Step 3: Synthesis of Intermediate 37-4

[0405] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 37-3 instead of intermediate 34-2 to obtain intermediate 37-4. ESI-MS: m / z=438 [M+H] + .

[0406] Synthesis Step 4: Synthesis of N-(2'-fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-4'-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A37)

[0407] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used to replace intermediate 1-4, and intermediate 37-4 was used to replace intermediate 34-3 to obtain compound A37. ESI-MS: m / z=591 [M+H] + .

[0408] Example 38: N-(2',4',6'-trifluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A38)

[0409]

[0410] Synthesis step 1: Synthesis of intermediate 38-1

[0411] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 37-3, and replace intermediate 1-10 with 2,4,6-trifluoroiodobenzene to obtain intermediate 38-1. ESI-MS: m / z=302[M+H] + .

[0412] Synthesis Step 2: Synthesis of Intermediate 38-2

[0413] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 38-1 instead of intermediate 34-2 to obtain intermediate 38-2. ESI-MS: m / z=406 [M+H] + .

[0414] Synthesis step 3: Synthesis of N-(2',4',6'-trifluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A38)

[0415] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used to replace intermediate 1-4, and intermediate 38-2 was used to replace intermediate 34-3 to obtain compound A38. ESI-MS: m / z=559 [M+H] + .

[0416] Example 39: N-(2',6'-difluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A39)

[0417]

[0418] Synthesis step 1: Synthesis of intermediate 39-1

[0419] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 37-3, and replace intermediate 1-10 with 2,6-difluoroiodobenzene to obtain intermediate 39-1. ESI-MS: m / z=284[M+H] + .

[0420] Synthesis Step 2: Synthesis of Intermediate 39-2

[0421] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 39-1 instead of intermediate 34-2 to obtain intermediate 39-2. ESI-MS: m / z=388[M+H] + .

[0422] Synthesis step 3: Synthesis of N-(2',6'-difluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A39)

[0423] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A39. ESI-MS: m / z=541[M+H] + .

[0424] Example 40: N-(4'-fluoro-2'-methoxy-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A40)

[0425]

[0426] Synthesis step 1: Synthesis of intermediate 40-1

[0427] Synthesis steps: Referring to step 9 of Example 1, intermediate 37-3 was used to replace intermediate 1-5, and 2-bromo-5-fluoroanisole was used to replace intermediate 1-10 to obtain intermediate 40-1. ESI-MS: m / z=296 [M+H] + .

[0428] Synthesis Step 2: Synthesis of Intermediate 40-2

[0429] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 40-1 instead of intermediate 34-2 to obtain intermediate 40-2. ESI-MS: m / z=400[M+H] + .

[0430] Synthesis step 3: Synthesis of N-(4'-fluoro-2'-methoxy-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A40)

[0431] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used to replace intermediate 1-4, and intermediate 40-2 was used to replace intermediate 34-3 to obtain compound A40. ESI-MS: m / z=553 [M+H] + .

[0432] Example 41: N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A41)

[0433]

[0434] Synthesis Step 1: Synthesis of Intermediate 41-1

[0435] Synthesis steps: Referring to step 9 of Example 1, intermediate 37-3 was used to replace intermediate 1-5, and 2-bromo-5-fluoroanisole was used to replace intermediate 1-10 to obtain intermediate 41-1. ESI-MS: m / z=285[M+H] + .

[0436] Synthesis Step 2: Synthesis of Intermediate 41-2

[0437] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 41-1 instead of intermediate 34-2 to obtain intermediate 41-2. ESI-MS: m / z=389 [M+H] + .

[0438] Synthesis step 3: Synthesis of N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A41)

[0439] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used to replace intermediate 1-4, and intermediate 41-2 was used to replace intermediate 34-3 to obtain compound A41. ESI-MS: m / z=542[M+H] + .

[0440] Example 42: N-(2'-fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-4'-(trifluoromethoxy)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A42)

[0441]

[0442] Synthesis Step 1: Synthesis of Intermediate 42-1

[0443] Synthesis steps: Referring to step 9 of Example 1, intermediate 37-3 was used to replace intermediate 1-5, and 1-bromo-2-fluoro-4-trifluoromethoxybenzene was used to replace intermediate 1-10 to obtain intermediate 42-1. ESI-MS: m / z=350 [M+H] + .

[0444] Synthesis Step 2: Synthesis of Intermediate 42-2

[0445] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 42-1 instead of intermediate 34-2 to obtain intermediate 42-2. ESI-MS: m / z=454 [M+H] + .

[0446] Synthesis step 3: Synthesis of N-(2'-fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-4'-(trifluoromethoxy)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A42)

[0447] Synthesis steps: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 15-3, and replace intermediate 34-3 with intermediate 42-2 to obtain compound A42. 1 H NMR(400MHz,Chloroform-d)δ8.32(d,J=7.6Hz,1H),8.08(s,1H),7.57-7.45(m,3H),7. 25(s,1H),7.16-7.05(m,3H),6.57(dd,J=7.5,2.6Hz,1H),4.13-3.91(m,4H),3.77-3.6 6(m,1H),3.09(d,J=12.1Hz,1H),2.99-2.77(m,3H),2.57(dt,J=8.3,3.5Hz,1H),2.03( s,1H),1.00(dd,J=8.0,2.4Hz,2H),0.85(dd,J=9.7,3.3Hz,2H).ESI-MS:m / z=607[M+H] + .

[0448] Example 43: N-(3-(2,6-difluoropyridin-3-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A43)

[0449]

[0450] Synthesis step 1: Synthesis of intermediate 43-1

[0451] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 37-3, and replace intermediate 1-10 with 3-bromo-2,6-difluoropyridine to obtain intermediate 43-1. ESI-MS: m / z=285[M+H] + .

[0452] Synthesis Step 2: Synthesis of Intermediate 43-2

[0453] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 43-1 instead of intermediate 34-2 to obtain intermediate 43-2. ​​ESI-MS: m / z=389 [M+H] + .

[0454] Synthesis step 3: Synthesis of N-(3-(2,6-difluoropyridin-3-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A43)

[0455] Synthesis steps: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 15-3, and replace intermediate 34-3 with intermediate 43-2 to obtain compound A43. 1 H NMR(400MHz,Chloroform-d)δ8.33(d,J=7.6Hz,1H),8.11-8.00(m,2H),7.57(t,J=1.8Hz,1H),7.47(q,J=1 .6Hz,1H),7.25(d,J=1.7Hz,1H),7.10(d,J=2.6Hz,1H),6.96(dd,J=8.1,2.9Hz,1H),6.58(dd,J=7.6,2.6H z,1H),4.05(qd,J=10.3,5.0Hz,2H),3.99-3.89(m,2H),3.70(td,J=11.2,2.9Hz,1H),3.09-3.00(m,1H),2 .95-2.76(m,3H),2.59(tt,J=8.0,4.8Hz,1H),1.24-1.19(m,2H),1.05-0.97(m,2H).ESI-MS:m / z=542[M+H] + .

[0456] Example 44: N-(2'-chloro-4'-fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A44)

[0457]

[0458] Synthesis Step 1: Synthesis of Intermediate 44-1

[0459] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 37-3, and replace intermediate 1-10 with 1-bromo-2-chloro-4-fluorobenzene to obtain intermediate 44-1. ESI-MS: m / z=300 [M+H] + .

[0460] Synthesis Step 2: Synthesis of Intermediate 44-2

[0461] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 44-1 instead of intermediate 34-2 to obtain intermediate 44-2. ESI-MS: m / z=404 [M+H] + .

[0462] Synthesis step 3: Synthesis of N-(2'-chloro-4'-fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A44)

[0463] Synthesis procedure: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 15-3, and replace intermediate 34-3 with intermediate 44-2 to obtain compound A44. 1 H NMR(400MHz,Chloroform-d)δ8.31(d,J=7.6Hz,1H),8.07(s,1H),7.54(s,1H),7 .39-7.32(m,2H),7.25(s,2H),7.15-7.01(m,3H),6.57(dd,J=7.9,2.6Hz,1H),4 .10-3.88(m,3H),3.74-3.64(m,1H),3.04(d,J=11.9Hz,1H),2.85(ddt,J=30.2, 22.2,10.7Hz,3H),1.27-1.18(m,3H),1.04-0.97(m,2H).ESI-MS:m / z=557[M+H] + .

[0464] Example 45: N-(4'-chloro-2'-fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A45)

[0465]

[0466] Synthesis Step 1: Synthesis of Intermediate 45-1

[0467] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 37-3, and replace intermediate 1-10 with 1-bromo-4-chloro-2-fluorobenzene to obtain intermediate 45-1. ESI-MS: m / z=300 [M+H] + .

[0468] Synthesis Step 2: Synthesis of Intermediate 45-2

[0469] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 45-1 instead of intermediate 34-2 to obtain intermediate 45-2. ESI-MS: m / z=404 [M+H] + .

[0470] Synthesis step 3: Synthesis of N-(4'-chloro-2'-fluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A45)

[0471] Synthesis steps: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 15-3, and replace intermediate 34-3 with intermediate 45-2 to obtain compound A45. 1 H NMR(400MHz,Chloroform-d)δ8.30(d,J=7.6Hz,1H),8.07(s,1H),7.55(t,J=1.8Hz,1H),7.48-7.37(m,2H),7.21(s,3H),7.10(d,J=2.7Hz,1H),6.5 6(dd,J=7.6,2.6Hz,1H),4.05-3.91(m,3H),3.76-3.63(m,2H),3.12-2.7 8(m,4H),1.23(s,3H),0.99(dd,J=7.8,2.3Hz,2H).ESI-MS:m / z=557[M+H] + .

[0472] Example 46: N-(3-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A46)

[0473]

[0474] Synthesis Step 1: Synthesis of Intermediate 46-1

[0475] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 37-3, and replace intermediate 1-10 with 5-bromo-2,2-difluoro-1,3-benzodioxole to obtain intermediate 46-1. ESI-MS: m / z=328 [M+H] + .

[0476] Synthesis Step 2: Synthesis of Intermediate 46-2

[0477] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 46-1 instead of intermediate 34-2 to obtain intermediate 46-2. ESI-MS: m / z=432 [M+H] + .

[0478] Synthesis step 3: Synthesis of N-(3-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A46)

[0479] Synthesis steps: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 15-3, and replace intermediate 34-3 with intermediate 46-2 to obtain compound A46. 1 H NMR(400MHz,Chloroform-d)δ8.33(d,J=7.6Hz,1H),8.10(s,1H),7.56(t,J=1.8Hz,1H),7.45(t,J=1.6Hz,1H),7.3 4-7.29(m,2H),7.21(t,J=1.8Hz,1H),7.17-7.13(m,1H),7.11(d,J=2.6Hz,1H),6.59(dd,J=7.6,2.6Hz,1H),4.15-4 .01(m,2H),4.00-3.91(m,2H),3.72(td,J=11.1,3.2Hz,1H),3.09(dd,J=12.2,2.4Hz,1H),3.01-2.87(m,2H),2.82 (dd,J=12.1,10.0Hz,1H),2.58(tt,J=8.0,4.8Hz,1H),1.04-0.97(m,2H),0.91-0.81(m,1H).ESI-MS:m / z=585[M+H] + .

[0480] Example 47: N-(2',4',5'-trifluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A47)

[0481]

[0482] Synthesis Step 1: Synthesis of Intermediate 47-1

[0483] Synthesis steps: Refer to step 9 of Example 1, replace intermediate 1-5 with intermediate 37-3, and replace intermediate 1-10 with 2,4,5-trifluorobromobenzene to obtain intermediate 47-1. ESI-MS: m / z=302[M+H] + .

[0484] Synthesis Step 2: Synthesis of Intermediate 47-2

[0485] Synthesis steps: Refer to step 2 of Example 34, and use intermediate 47-1 instead of intermediate 34-2 to obtain intermediate 47-2. ESI-MS: m / z=432 [M+H] + .

[0486] Synthesis step 3: Synthesis of N-(2',4',5'-trifluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A47)

[0487] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used to replace intermediate 1-4, and intermediate 47-2 was used to replace intermediate 34-3 to obtain compound A47. ESI-MS: m / z=559 [M+H] + .

[0488] Example 48: N-(5-(5-(2-(dimethylamino)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A48)

[0489]

[0490] Synthesis steps Referring to step 3 of Example 34, intermediate 28-2 was used instead of intermediate 1-4 to obtain compound A48, 1 HNMR(400MHz,Chloroform-d)δ8.14(d,J=7.5Hz,1H),8.05(s,1H),7.65(t,J=1.9Hz ,1H),7.50-7.42(m,2H),7.34-7.30(m,2H),6.96(dddd,J=19.4,11.1,8.6,2.6Hz,2H ),6.31(dd,J=7.4,2.4Hz,1H),4.86(s,2H),3.59(s,2H),3.06(d,J=4.5Hz,6H),2.56 (tt,J=8.0,4.6Hz,1H),1.21-1.17(m,2H),0.90-0.87(m,2H).ESI-MS:m / z=513[M+H] + .

[0491] Example 49: N-(2',4'-difluoro-5-(5-(piperidin-4-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A49)

[0492]

[0493] Synthesis step 1: Synthesis of intermediate 49-1

[0494] Synthesis steps: Referring to step 3 of Example 1, intermediate 34-3 was used instead of intermediate 1-3 to obtain compound intermediate 49-1, ESI-MS: m / z=488 [M+H] + .

[0495] Synthesis step 2: Preparation of N-(2',4'-difluoro-5-(5-(piperidin-4-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A49)

[0496] Synthesis steps Referring to step 3 of Example 34, intermediate 11-1 is used instead of intermediate 1-4, and intermediate 49-1 is used instead of intermediate 34-3 to obtain compound A49. 1 H NMR(400MHz, Methanol-d4)δ8.41(d,J=7.6Hz,1H),8.18(s,1H),7.71(t,J=1.8Hz,1H),7.62-7.53(m,1H), 7.49(q,J=1.7Hz,1H),7.27(d,J=2.6Hz,1H),7.20(q,J=1.8Hz,1H),7.11-7.03(m,2H),6.64(dd,J=7.6,2. 6Hz,1H),4.03(d,J=6.2Hz,2H),3.42(d,J=12.9Hz,2H),3.03(td,J=12.9,3.0Hz,2H),2.63(td,J=8.1,4.0 Hz,1H),2.32-2.01(m,3H),1.73-1.43(m,2H),1.10-1.02(m,2H),1.01-0.93(m,2H).ESI-MS:m / z=539[M+H] + .

[0497] Example 50: (S)-N-(2',4'-difluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A50)

[0498]

[0499] Synthesis steps Referring to step 3 of Example 34, intermediate 17-3 was used instead of intermediate 1-4 to obtain compound A50, 1HNMR(400MHz, Methanol-d4)δ8.44-8.39(m,1H),8.18(s,1H),7.67(t,J=1.8Hz,1H),7.59(td,J=8.8,6.3Hz,1H),7. 50(q,J=1.6Hz,1H),7.26(q,J=1.4Hz,2H),7.14-7.05(m,2H),6.68(dd,J=7.6,2.6Hz,1H),4.15-4.08(m,2H),3.94(d dq,J=10.8,8.0,2.5Hz,2H),3.69(ddd,J=11.8,9.2,5.1Hz,1H),3.07(dd,J=12.6,2.4Hz,1H),2.91-2.84(m,2H),2.7 9(dd,J=12.5,10.7Hz,1H),2.66(tt,J=8.0,4.8Hz,1H),1.14-1.07(m,2H),1.04-0.96(m,2H).ESI-MS:m / z=541[M+H] + .

[0500] Example 51: (R)-N-(2',4'-difluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A51)

[0501]

[0502] Synthesis steps Referring to step 3 of Example 34, intermediate 18-3 was used instead of intermediate 1-4 to obtain compound A51, 1HNMR (400MHz, Methanol-d4) δ8.39(dd,J=7.7,1.2Hz,1H),8.15(d,J=1.0Hz,1H),7.64(t,J=1.8Hz,1H),7.56(td,J=8.9,6.5H z,1H),7.47(q,J=1.5Hz,1H),7.23(t,J=2.0Hz,2H),7.07(ddt,J=13.5,8.2,2.7Hz,2H),6.65(dt,J=7.6,2.0Hz,1H),4.08(d,J =4.5Hz,2H),3.95-3.81(m,2H),3.64(ddd,J=11.6,9.9,4.3Hz,1H),3.00(dd,J=12.6,2.4Hz,1H),2.82(dd,J=9.8,3.3Hz,2H), 2.72(dd,J=12.5,10.6Hz,1H),2.63(ddd,J=8.0,4.9,3.2Hz,1H),1.11-1.04(m,2H),1.01-0.93(m,2H).ESI-MS:m / z=541[M+H] + .

[0503] Example 52: N-(5-(5-(((1r,4r)-4-aminocyclohexyl)oxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A52)

[0504]

[0505] Synthesis Step 1: Synthesis of Intermediate 52-2

[0506] Synthesis procedure: Refer to step 1 of Example 10, and use cis-N-BOC-4-aminocyclohexanol instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 52-2, which is directly used in the next step.

[0507] Synthesis Step 2: Synthesis of Intermediate 52-3

[0508] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used to replace 5-2, and intermediate 52-2 was used to replace 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 52-3. Yield: 63%; ESI-MS: m / z=332[M+1] + .

[0509] Synthesis step 3: Preparation of N-(5-(5-(((1r,4r)-4-aminocyclohexyl)oxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A52)

[0510] Synthesis steps Referring to step 3 of Example 34, intermediate 52-3 was used instead of intermediate 1-4 to obtain compound A52, 1 HNMR(400MHz,Chloroform-d)δ8.31(d,J=7.6Hz,1H),8.08(s,1H),7.57(t,J=1.8Hz,1H),7.50- 7.38(m,2H),7.20-7.11(m,2H),6.99-6.86(m,2H),6.46(dd,J=7.6,2.6Hz,1H),4.46-4.18(m,1H ),2.81(d,J=10.3Hz,1H),2.57(tt,J=8.0,4.8Hz,1H),2.28-2.12(m,2H),2.02-1.89(m,2H),1. 58-1.45(m,2H),1.42-1.31(m,2H),1.22-1.16(m,2H),1.02-0.94(m,2H).ESI-MS:m / z=541[M+H] + .

[0511] Example 53: N-(5-(5-(((1s,4s)-4-aminocyclohexyl)oxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A53)

[0512]

[0513] Synthesis steps Referring to step 3 of Example 34, intermediate 26-3 was used instead of intermediate 1-4 to obtain compound A53, 1HNMR(400MHz,Chloroform-d)δ8.42(d,J=7.5Hz,1H),8.19(s,1H),7.68(t,J=1.9Hz,1 H),7.55-7.50(m,2H),7.46(td,J=8.6,6.4Hz,1H),7.41(d,J=1.5Hz,1H),7.02-6.89(m ,2H),6.51(dd,J=7.7,2.4Hz,1H),4.47(s,1H),2.67(tt,J=8.2,4.7Hz,1H),2.00(s,4H ),1.75(d,J=15.4Hz,4H),1.00-0.95(m,2H),0.91-0.84(m,2H).ESI-MS:m / z=541[M+H] + .

[0514] Example 54: N-(5-(5-(((1r,4r)-4-aminocyclohexyl)methoxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A54)

[0515]

[0516] Synthesis Step 1: Synthesis of Intermediate 54-2

[0517] Synthesis procedure Referring to step 1 of Example 10, tert-butyl trans-(4-hydroxymethyl)cyclohexylcarbamate was used instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 54-2, which was directly used in the next step.

[0518] Synthesis Step 2: Synthesis of Intermediate 54-3

[0519] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used to replace 5-2, and intermediate 54-2 was used to replace 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 54-3, yield: 63%; ESI-MS: m / z=346[M+1] + .

[0520] Synthesis step 3: Preparation of N-(5-(5-(((1r,4r)-4-aminocyclohexyl)methoxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A54)

[0521] Synthesis steps Referring to step 3 of Example 34, intermediate 54-3 was used instead of intermediate 1-4 to obtain compound A54, 1HNMR(400MHz,Chloroform-d)δ8.30(dd,J=7.6,0.6Hz,1H),8.06(s,1H),7.56(t,J=1.8Hz,1H),7 .47-7.39(m,2H),7.20(q,J=1.6Hz,1H),7.08(d,J=2.6Hz,1H),7.00-6.86(m,2H),6.49(dd,J=7.6 ,2.5Hz,1H),3.82(d,J=6.3Hz,2H),2.72-2.61(m,3H),2.59-2.53(m,1H),1.90(d,J=5.3Hz,4H),1 .83-1.72(m,1H),1.20-1.17(m,2H),1.15-1.09(m,4H),1.00-0.95(m,2H).ESI-MS:m / z=553[M+H] + .

[0522] Example 55: N-(2',4'-difluoro-5-(5-((4-methylmorpholin-2-yl)methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A55)

[0523]

[0524] Synthesis Step 1: Synthesis of Intermediate 55-2

[0525] The raw material 55-1 (868 mg, 4 mmol) was dissolved in DCM (8 mL), and 4 mL of trifluoroacetic acid was added dropwise under ice bath. After the addition, the mixture was moved to room temperature for 3 h. The solvent was removed by distillation under reduced pressure and the mixture was directly used in the next step.

[0526] Synthesis Step 2: Synthesis of Intermediate 55-3

[0527] Synthesis steps: Referring to step 4 of Example 10, intermediate 55-2 was used instead of 10-4 to obtain intermediate 55-3, yield: 63%; ESI-MS: m / z=132[M+1] + .

[0528] Synthesis Step 3: Synthesis of Intermediate 55-4

[0529] Synthesis procedure: Referring to step 1 of Example 10, intermediate 55-3 was used instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 55-4, which was directly used in the next step.

[0530] Synthesis Step 4: Synthesis of Intermediate 55-5

[0531] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used to replace 5-2, and intermediate 55-4 was used to replace 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 55-5, yield: 43%; ESI-MS: m / z=262[M+1] + .

[0532] Synthesis Step 5: Preparation of N-(2',4'-difluoro-5-(5-((4-methylmorpholin-2-yl)methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A55)

[0533] Synthesis steps: Referring to step 3 of Example 34, intermediate 55-5 was used instead of intermediate 1-4 to obtain compound A55, ESI-MS: m / z=555 [M+H] + .

[0534] Example 56: N-(2',4'-difluoro-5-(5-(pyrrolidin-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A56)

[0535]

[0536] Synthesis Step 1: Synthesis of Intermediate 56-2

[0537] Synthesis procedure: Referring to step 1 of Example 10, 1-Boc-3-hydroxymethylpyrrolidine was used instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 56-2, which was directly used in the next step.

[0538] Synthesis Step 2: Synthesis of Intermediate 56-3

[0539] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used instead of 5-2, and intermediate 56-2 was used instead of 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 56-3, yield: 51%; ESI-MS: m / z=318[M+1] + .

[0540] Synthesis step 3: Preparation of N-(2',4'-difluoro-5-(5-(pyrrolidine-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A56)

[0541] Synthesis steps Referring to step 3 of Example 34, intermediate 56-3 was used instead of intermediate 1-4 to obtain compound A56, 1HNMR(400MHz,Chloroform-d)δ8.32(d,J=7.5Hz,1H),8.13(s,1H),7.69(t,J=1.9Hz,1H),7.51-7.42(m ,3H),7.35(d,J=2.6Hz,1H),7.01-6.88(m,2H),6.52(dd,J=7.6,2.6Hz,1H),4.37(t,J=9.5Hz,1H),4.0 4(dd,J=9.6,5.6Hz,1H),3.48(td,J=10.5,8.8,6.3Hz,3H),3.37-3.28(m,1H),2.92(s,1H),2.60(tt,J =8.0,4.8Hz,1H),2.33-2.19(m,1H),2.09(d,J=12.7Hz,1H),1.27-1.10(m,4H).ESI-MS:m / z=525[M+H] + .

[0542] Example 57: N-(5-(5-((1-(dimethylamino)propan-2-yl)oxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A57)

[0543]

[0544] Synthesis Step 1: Synthesis of Intermediate 57-2

[0545] N, N-dimethylisopropanolamine (2 g) was dissolved in 30 mL of acetone, and 2.6 g of methylsulfonyl chloride was added dropwise under ice bath conditions, stirring was maintained, and the temperature was naturally restored. After 1 hour, the reaction was completed, and the solvent was removed by distillation under reduced pressure. Dichloromethane was added, and the mixture was filtered. After being rinsed three times with dichloromethane, the filter cake was taken out and dried, and the solid was taken out. 2 g of a white powdery solid (57-2) was obtained.

[0546] Synthesis Step 2: Synthesis of Intermediate 57-3

[0547] Synthesis steps: Refer to step 2 of Example 5, use intermediate 12-5 instead of 5-2, and use intermediate 57-2 instead of 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 57-3, yield: 31%; ESI-MS: m / z=285[M+H]+.

[0548] Synthesis step 3: Preparation of N-(5-(5-((1-(dimethylamino)propane-2-yl)oxy)pyrazolo[1,5-a]pyridin-3-yl)-2',4'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A57)

[0549] Synthesis steps: Referring to step 3 of Example 34, intermediate 57-3 was used instead of intermediate 1-4 to obtain compound A57, ESI-MS: m / z=527 [M+H] + .

[0550] Example 58: (S)-N-(2',4'-difluoro-5-(5-(pyrrolidin-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A58)

[0551]

[0552] Synthesis Step 1: Synthesis of Intermediate 58-2

[0553] Synthesis procedure: Referring to step 1 of Example 10, (S)-1-Boc-3-hydroxymethylpyrrolidine was used instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 58-2, which was directly used in the next step.

[0554] Synthesis Step 2: Synthesis of Intermediate 58-3

[0555] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used instead of 5-2, and intermediate 58-2 was used instead of 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 58-3, yield: 51%; ESI-MS: m / z=318[M+1] + .

[0556] Synthesis step 3: Preparation of (S)-N-(2',4'-difluoro-5-(5-(pyrrolidine-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A58)

[0557] Synthesis steps Referring to step 3 of Example 34, intermediate 58-3 was used instead of intermediate 1-4 to obtain compound A58, 1HNMR(400MHz,Chloroform-d)δ8.31(d,J=7.5Hz,1H),8.12(s,1H),7.68(t,J=2.0Hz,1H),7.50-7.43(m ,3H),7.33(d,J=2.6Hz,1H),7.00-6.87(m,2H),6.51(dd,J=7.6,2.6Hz,1H),4.36(t,J=9.5Hz,1H),4.0 3(dd,J=9.6,5.6Hz,1H),3.47(td,J=10.4,8.8,6.4Hz,3H),3.36-3.25(m,1H),2.93(s,1H),2.61(tt,J =8.0,4.8Hz,1H),2.32-2.18(m,1H),2.08(d,J=12.8Hz,1H),1.26-1.10(m,4H).ESI-MS:m / z=525[M+H] + .

[0558] Example 59: (R)-N-(2',4'-difluoro-5-(5-(pyrrolidin-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A59)

[0559]

[0560] Synthesis Step 1: Synthesis of Intermediate 59-2

[0561] Synthesis procedure: Referring to step 1 of Example 10, (R)-1-Boc-3-hydroxymethylpyrrolidine was used instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 59-2, which was directly used in the next step.

[0562] Synthesis Step 2: Synthesis of Intermediate 59-3

[0563] Synthesis steps: Referring to step 2 of Example 5, intermediate 12-5 was used instead of 5-2, and intermediate 59-2 was used instead of 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 59-3, yield: 51%; ESI-MS: m / z=318[M+1] + .

[0564] Synthesis step 3: Preparation of (R)-N-(2',4'-difluoro-5-(5-(pyrrolidine-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A59)

[0565] Synthesis steps: Referring to step 3 of Example 34, intermediate 59-3 was used instead of intermediate 1-4 to obtain compound A59, ESI-MS: m / z=525 [M+H] + .

[0566] Example 60: N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(2-(dimethylamino)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A60)

[0567]

[0568] Synthesis steps Referring to step 3 of Example 34, intermediate 28-2 was used to replace intermediate 1-4, and intermediate 41-2 was used to replace intermediate 34-3 to obtain compound A60, ESI-MS: m / z=514 [M+H] + .

[0569] Example 61: (R)-N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(pyrrole-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A61)

[0570]

[0571] Synthesis steps Referring to step 3 of Example 34, intermediate 59-3 was used to replace intermediate 1-4, and intermediate 41-2 was used to replace intermediate 34-3 to obtain compound A61. 1 H NMR(400MHz,Chloroform-d)δ8.47(d,J=2.4Hz,1H),8.32(d,J=7.5Hz,1H),8.20(s,1H),7.89(p,J=1.4Hz, 2H),7.78(t,J=1.9Hz,1H),7.39-7.28(m,2H),6.50(dd,J=7.6,2.6Hz,1H),4.42(t,J=9.7Hz,1H),4.05(dd ,J=9.6,5.2Hz,1H),3.61-3.46(m,3H),3.42(d,J=11.7Hz,1H),2.96(s,1H),2.61(tt,J=8.0,4.8Hz,1H),2 .36-2.23(m,1H),2.22-2.12(m,1H),1.22-1.16(m,2H),0.91(dd,J=8.2,4.0Hz,2H).ESI-MS:m / z=526[M+H] + .

[0572] Example 62: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(pyrrole-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A62)

[0573]

[0574] Synthesis steps Referring to step 3 of Example 34, intermediate 58-3 was used to replace intermediate 1-4, and intermediate 41-2 was used to replace intermediate 34-3 to obtain compound A62, ESI-MS: m / z=526 [M+H] + .

[0575] Example 63: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A63)

[0576]

[0577] Synthesis steps Referring to step 3 of Example 34, intermediate 17-3 was used instead of intermediate 1-4, and intermediate 41-2 was used instead of intermediate 34-3 to obtain compound A63. 1 H NMR(400MHz,Chloroform-d)δ8.47(dt,J=2.4,0.8Hz,1H),8.33(dd,J=7.6,0.6Hz,1H),8.15-8.12(m,1H),7.93(q,J=1.6Hz,1H),7.6 9(t,J=1.8Hz,1H),7.59(q,J=1.6Hz,1H),7.35(ddd,J=10.4,7.9,2.4Hz,1H),7.14(d,J=2.6Hz,1H),6.59(dd,J=7.6,2.6Hz,1H),4.12 -4.00(m,2H),4.00-3.91(m,2H),3.71(td,J=11.2,2.9Hz,1H),3.06(d,J=11.8Hz,1H),2.96(td,J=11.7,11.0,3.2Hz,1H),2.88(d,J= 11.9Hz, 1H), 2.82 (dd, J=12.1, 10.2Hz, 1H), 2.58 (tt, J=8.0, 4.8Hz, 1H), 1.04-0.97 (m, 2H), 0.91-0.81 (m, 1H). ESI-MS: m / z=542[M+H] + .

[0578] Example 64: (R)-N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A64)

[0579]

[0580] Synthesis steps Referring to step 3 of Example 34, intermediate 18-3 was used to replace intermediate 1-4, and intermediate 41-2 was used to replace intermediate 34-3 to obtain compound A64. 1 H NMR (400MHz, DMSO-d6) δ8.71-8.60(m,2H),8.29(s,1H),8.15-8.06(m,1H),7.82(q,J=1.7Hz,1H),7.65(d,J=1.6Hz,2H),7.18(d,J=2.7Hz,1H),6.71 (dd,J=7.6,2.6Hz,1H),4.05(d,J=5.1Hz,2H),3.84-3.71(m,2H),3.49(d, J=3.6Hz,1H),2.95-2.52(m,5H),1.04-0.95(m,4H).ESI-MS:m / z=542[M+H] + .

[0581] Example 65: (S)-N-(2',6'-difluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A65)

[0582]

[0583] Synthesis steps Referring to step 3 of Example 34, intermediate 17-3 was used instead of intermediate 1-4, and intermediate 39-2 was used instead of intermediate 34-3 to obtain compound A65. 1H NMR(400MHz,Chloroform-d)δ8.31(d,J=7.6Hz,1H),8.07(s,1H),7.57(t,J=1.9Hz,1H),7.47-7.42(m ,1H),7.31(tt,J=8.4,6.3Hz,1H),7.20(q,J=1.6Hz,1H),7.11(d,J=2.7Hz,1H),7.00(t,J=7.9Hz,2H), 6.55(dd,J=7.6,2.7Hz,1H),4.09-3.97(m,2H),3.97-3.83(m,2H),3.74-3.65(m,1H),3.09-2.98(m,1 H),2.95-2.75(m,3H),2.62-2.50(m,1H),1.22-1.17(m,2H),1.03-0.93(m,2H).ESI-MS: m / z=541[M+H] + .

[0584] Example 66: (R)-N-(2',6'-difluoro-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A66)

[0585]

[0586] Synthesis steps Referring to step 3 of Example 34, intermediate 18-3 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A66. 1 H NMR(400MHz,Chloroform-d)δ8.31(d,J=7.6Hz,1H),8.07(s,1H),7.56(d,J=1.9Hz,1H),7.43(s ,1H),7.35-7.28(m,1H),7.20(s,1H),7.11(d,J=2.7Hz,1H),6.99(t,J=7.9Hz,2H),6.55(dt,J= 7.6,2.2Hz,1H),4.08-3.99(m,2H),3.97-3.89(m,2H),3.72-3.66(m,1H),3.07-2.98(m,1H),2. 93-2.77(m,3H),2.61-2.51(m,1H),1.21-1.17(m,2H),1.05-0.95(m,2H).ESI-MS:m / z=541[M+H] + .

[0587] Example 67: (S)-N-(2',6'-difluoro-5-(5-(pyrrole-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A67)

[0588]

[0589] Synthesis steps Referring to step 3 of Example 34, intermediate 58-3 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A67. 1 H NMR(400MHz,Chloroform-d)δ8.28(d,J=7.5Hz,1H),8.05(s,1H),7.57(t,J=1.9Hz,1H),7.43-7.39(m,1H),7.29 (ddd,J=8.5,6.3,2.1Hz,1H),7.20(t,J=1.7Hz,1H),7.12(d,J=2.6Hz,1H),6.98(t,J=7.9Hz,2H),6.46(dd,J=7.6 ,2.6Hz,1H),4.00-3.90(m,2H),3.20-3.12(m,1H),3.10-3.02(m,1H),3.01-2.91(m,1H),2.90-2.81(m,1H),2.68 -2.60(m,1H),2.60-2.51(m,1H),2.06-1.95(m,1H),1.19-1.15(m,2H),0.99-0.91(m,2H).ESI-MS:m / z=525[M+H] + .

[0590] Example 68: (R)-N-(2',6'-difluoro-5-(5-(pyrrole-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A68)

[0591]

[0592] Synthesis steps Referring to step 3 of Example 34, intermediate 59-3 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A68. 1H NMR(400MHz,Chloroform-d)δ8.28(d,J=7.6Hz,1H),8.07(s,1H),7.62(t,J=1.8Hz,1H),7.44-7.38(m,1H),7.33 -7.26(m,2H),7.19(d,J=2.6Hz,1H),6.98(t,J=7.9Hz,2H),6.47(dd,J=7.5,2.6Hz,1H),4.10(dd,J=9.4,7.3Hz,1 H),3.97(dd,J=9.4,6.9Hz,1H),3.32-3.22(m,1H),3.21-3.08(m,2H),3.05-2.96(m,1H),2.79-2.65(m,1H),2.63 -2.50(m,1H),2.14-2.04(m,1H),1.82-1.70(m,1H),1.22-1.15(m,2H),0.99-0.91(m,2H).ESI-MS:m / z=525[M+H] + .

[0593] Example 69: N-(5-(5-(2-(dimethylamino)methoxy)pyrazolo[1,5-a]pyridin-3-yl)-2',6'-difluoro-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A69)

[0594]

[0595] Synthesis steps Referring to step 3 of Example 34, intermediate 28-2 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A69, ESI-MS: m / z=513 [M+H] + .

[0596] Example 70: (S)-N-(2',6'-difluoro-5-(6-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-33-yl)cyclopropanesulfonamide (A70)

[0597]

[0598] Synthesis Step 1: Synthesis of Intermediate 70-1

[0599] Synthesis steps: Referring to step 2 of Example 5, intermediate 6-2 was used instead of 5-2, and intermediate 17-2 was used instead of 4-(2-bromoethyl)morpholine hydrobromide to obtain intermediate 70-1, yield: 57%; ESI-MS: m / z=334[M+1] + .

[0600] Synthesis step 2: Synthesis of (S)-N-(2',6'-difluoro-5-(6-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-33-yl)cyclopropanesulfonamide (A70)

[0601] Synthesis steps Referring to step 3 of Example 34, intermediate 70-1 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A70, ESI-MS: m / z=541 [M+H] + .

[0602] Example 71: N-(2',4'-difluoro-5-(6-(2-morpholinylethoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A71)

[0603]

[0604] Synthesis steps Referring to step 3 of Example 34, intermediate 6-3 was used instead of intermediate 1-4 to obtain compound A71, 1 HNMR(400MHz,Chloroform-d)δ8.12(d,J=2.1Hz,1H),8.07(s,1H),7.72(d,J=9.7Hz ,1H),7.49(d,J=1.8Hz,2H),7.47-7.41(m,1H),7.28-7.26(m,1H),7.06(dd,J=9.7, 2.2Hz,1H),7.01-6.90(m,2H),4.34-4.07(m,2H),3.87-3.70(m,4H),3.05-2.80(m, 2H),2.78-2.30(m,5H),1.03-0.99(m,2H),0.87-0.83(m,2H).ESI-MS:m / z=555[M+H] + .

[0605] Example 72: N-(2',4'-difluoro-5-(6-(2-morpholinylethoxy)pyrazolo[1,5-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A72)

[0606]

[0607] Synthesis steps: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 6-3, replace intermediate 34-3 with intermediate 39-2, to obtain compound A72, 1H NMR(400MHz,Chloroform-d)δ8.12(d,J=2.1Hz,1H),8.07(s,1H),7.72(d,J=9.7Hz, 1H),7.49(d,J=1.8Hz,2H),7.47-7.41(m,1H),7.28-7.26(m,1H),7.06(dd,J=9.7,2 .2Hz,1H),7.01-6.90(m,2H),4.34-4.07(m,2H),3.87-3.70(m,4H),3.05-2.80(m,2 H),2.78-2.30(m,5H),1.03-0.99(m,2H),0.87-0.83(m,2H).ESI-MS:m / z=555[M+H] + .

[0608] Example 73: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A73)

[0609]

[0610] Synthesis steps: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 70-1, replace intermediate 34-3 with intermediate 41-2, to obtain compound A73, ESI-MS: m / z=542 [M+H] + .

[0611] Example 74: N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(2-morpholinoethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A74)

[0612]

[0613] Synthesis steps: Refer to step 3 of Example 34, replace intermediate 1-4 with intermediate 6-3, replace intermediate 34-3 with intermediate 41-2, to obtain compound A74, ESI-MS: m / z=556 [M+H] + .

[0614] Example 75: N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(2-morpholinoethoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A75)

[0615]

[0616] Synthesis Step 1: Synthesis of Intermediate 75-1

[0617] Synthesis steps: Referring to step 2 of Example 5, intermediate 23-2 was used instead of intermediate 5-2 to obtain intermediate 75-1, ESI-MS: m / z=248 [M+H] + .

[0618] Synthesis step 2: Preparation of N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(2-morpholinoethoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A75)

[0619] Synthesis steps Referring to step 3 of Example 34, intermediate 75-1 was used to replace intermediate 1-4, and intermediate 41-2 was used to replace intermediate 34-3 to obtain compound A75, ESI-MS: m / z=556 [M+H] + .

[0620] Example 76: N-(2',6'-difluoro-5-(7-(2-morpholinylethoxy)imidazo[1,2-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A76)

[0621]

[0622] Synthesis steps Referring to step 3 of Example 34, intermediate 75-1 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A76, ESI-MS: m / z=555 [M+H] + .

[0623] Example 77: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A77)

[0624]

[0625] Synthesis Step 1: Synthesis of Intermediate 77-1

[0626] Synthesis steps: Referring to step 2 of Example 5, intermediate 23-2 was used instead of intermediate 5-2 to obtain intermediate 77-1, ESI-MS: m / z=334 [M+H] + .

[0627] Synthesis step 2: Preparation of (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A77)

[0628] Synthesis steps Referring to step 3 of Example 34, intermediate 77-1 was used instead of intermediate 1-4, and intermediate 41-2 was used instead of intermediate 34-3 to obtain compound A77. 1 H NMR (400MHz, Chloroform-d) δ8.43(d,J=2.4Hz,1H),8.22(d,J=7.6Hz,1H),7.91(d,J=1.7Hz,1H),7.84(t,J=1. 3Hz,1H),7.63(s,1H),7.53(t,J=1.8Hz,1H),7.33(ddd,J=10.4,7.8,2.4Hz,1H),7.00(d,J=2.5Hz,1H),6.60(dd ,J=7.5,2.5Hz,1H),3.99-3.90(m,3H),3.70(d,J=2.9Hz,1H),3.65(s,1H),3.02(dd,J=12.2,2.4Hz,1H),2.96- 2.73(m,3H),2.66-2.56(m,1H),1.23(dt,J=5.8,3.7Hz,3H),0.98(tt,J=6.6,3.3Hz,2H).ESI-MS:m / z=542[M+H] + .

[0629] Example 78: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)methanesulfonamide (A78)

[0630]

[0631] Synthesis Step 1: Synthesis of Intermediate 78-1

[0632] Synthesis steps: Referring to step 2 of Example 34, methanesulfonyl chloride was used instead of cyclopropanesulfonyl chloride, and intermediate 41-1 was used instead of intermediate 34-2 to obtain intermediate 78-1, ESI-MS: m / z=363 [M+H] + .

[0633] Synthesis step 2: Preparation of (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)methanesulfonamide (A78)

[0634] Synthesis steps Referring to step 3 of Example 34, intermediate 17-3 was used instead of intermediate 1-4, and intermediate 78-1 was used instead of intermediate 34-3 to obtain compound A78, ESI-MS: m / z=516 [M+H] + .

[0635] Example 79: (S)-N-(2',6'-difluoro-5-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A79)

[0636]

[0637] Synthesis steps Referring to step 3 of Example 34, intermediate 77-1 was used to replace intermediate 1-4, and intermediate 39-2 was used to replace intermediate 34-3 to obtain compound A79, ESI-MS: m / z=541 [M+H] + .

[0638] Example 80: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(pyrrole-3-methoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A80)

[0639]

[0640] Synthesis step 1: Synthesis of intermediate 80-1

[0641] Synthesis steps: Referring to step 2 of Example 5, intermediate 23-2 was used instead of intermediate 5-2 to obtain intermediate 80-1, ESI-MS: m / z=318 [M+H] + .

[0642] Synthesis Step 2 Preparation of (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(pyrrole-3-methoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)cyclopropanesulfonamide (A80)

[0643] Synthesis steps Referring to step 3 of Example 34, intermediate 80-1 was used to replace intermediate 1-4, and intermediate 41-2 was used to replace intermediate 34-3 to obtain compound A80, ESI-MS: m / z=526 [M+H] + .

[0644] Example 81: (S)-N-(2',4'-difluoro-5-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)cyclopropanesulfonamide (A81)

[0645]

[0646] Synthesis steps: Referring to step 3 of Example 34, intermediate 77-1 was used instead of intermediate 1-4 to obtain compound A81, ESI-MS: m / z=541 [M+H] + .

[0647] Example 82: 2-(Difluoromethoxy)-6-methoxy-4-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)benzonitrile (A93)

[0648]

[0649] Synthesis Step 1: Synthesis of Intermediate 93-2

[0650] Add 4-bromo-2,6-difluorobenzoic acid (5.25 g) and tert-butylamine (1.62 g) to an eggplant-shaped bottle, then add 120 mL of acetonitrile to fully dissolve the raw materials. Then add HATU (9.13 g) and DIEA (8.52 g) at 0°C, stir for 10 min, and then react at room temperature for 4 h. After the reaction is completed, extract with DCM (50 mL) / water 5 times and combine the organic phases, and dry the organic phases with anhydrous sodium sulfate. Spin dry the organic phase to obtain 5.25 g (93-2) of the product as a white solid. ESI-MS: m / z=292[M+H] + .

[0651] Synthesis Step 2: Synthesis of Intermediate 93-3

[0652] Add intermediate 93-2 (5.25 g) and sodium methoxide (7.87 g) to an eggplant-shaped bottle, then add 100 mL of methanol to fully dissolve the raw materials, and react at 85°C overnight. After the reaction is completed, adjust the pH to 6-8 with 6N hydrochloric acid aqueous solution, then extract with DCM (60 mL) / water 3 times, combine the organic phases and dry with anhydrous sodium sulfate. Spin the organic phase and purify by column chromatography (PE: EA = 10: 1-8: 1-5: 1) to obtain 4.95 g (93-3) of a white solid. ESI-MS: m / z = 317 [M+H] + .

[0653] Synthesis Step 3: Synthesis of Intermediate 93-4

[0654] The intermediate 93-3 (4.95 g) was dissolved in 40 mL of DCM, and 20 mL of boron trichloride (1 M DCM solution) was added dropwise at -20 ° C, and then the reaction was continued at -20 ° C for 2.5 h. After the reaction was completed, 50 mL of water was added to quench the reaction, and the pH was adjusted to 7-8 with a saturated NaHCO3 solution. The organic phases were combined and dried over anhydrous sodium sulfate after extraction with DCM (50 mL) / water for 3 times. The organic phase was spin-dried and purified by column chromatography (PE: EA = 8: 1) to obtain 2.50 g of the product as a white solid (93-4). ESI-MS: m / z = 303 [M + H] + .

[0655] Synthesis Step 4: Synthesis of Intermediate 93-5

[0656] The intermediate 93-4 (2.50 g) was dissolved in a solution of acetonitrile: water = 1: 1 (V: V), KOH (4.65 g) was slowly added at 0°C, and after stirring for 15 minutes, diethyl bromofluoromethylphosphonate was added dropwise, and then reacted at 40°C for 7 hours. After the reaction was completed, the pH was adjusted to 7 with 6N hydrochloric acid aqueous solution, and then extracted with DCM (50 mL) / water three times, and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was spin-dried and purified by column chromatography (PE: EA = 10: 1 ~ 8: 1 ~ 5: 1) to obtain the product as a white solid 2.85 g (93-5). ESI-MS: m / z = 353 [M + H] + .

[0657] Synthesis Step 5: Synthesis of Intermediate 93-6

[0658] Dissolve the intermediate 93-5 (2.85 g) in 25 mL of phosphorus oxychloride and react at 110°C overnight. After the reaction is completed, cool the reaction solution to room temperature and slowly dropwise add 100 mL of ice water. Extract with DCM (50 mL) / water three times and combine the organic phases, which are dried over anhydrous sodium sulfate. Spin dry the organic phase and purify by column chromatography (PE:EA=30:1~20:1) to obtain 1.7 g (93-6) of a white solid product. ESI-MS: m / z=278[M+H] + .

[0659] Synthesis step 6: Synthesis of 2-(difluoromethoxy)-6-methoxy-4-(5-(morpholin-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)benzonitrile (A93)

[0660] Synthesis steps Referring to step 3 of Example 34, intermediate 15-3 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A93, ESI-MS: m / z=431 [M+H] + .

[0661] Example 83: 2-(Difluoromethoxy)-4-(7-((2-hydroxy-2-methylpropyl)amino)imidazo[1,2-a]pyridin-3-yl)-6-methoxybenzonitrile (A94)

[0662]

[0663] Synthesis Step 1: Synthesis of Intermediate 94-1

[0664] Synthesis steps Referring to step 3 of Example 34, intermediate 1-2 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain intermediate 94-1, ESI-MS: m / z=394 [M+H]+ .

[0665] Synthesis step 2: 2-(difluoromethoxy)-4-(7-((2-hydroxy-2-methylpropyl)amino)imidazo[1,2-a]pyridin-3-yl)-6-methoxybenzonitrile (A94)

[0666] Synthesis steps: Referring to step 2 of Example 1, intermediate 1-2 is used to replace intermediate 1-4, and intermediate 93-6 is used to replace 1-tert-butyloxycarbonyl-4-aminomethylpiperidine to obtain compound A94. 1 H NMR (400MHz, Methanol-d4) δ8.28 (d, J = 7.5 Hz, 1H), 7.57 (s, 1H), 7.32-6.91 (m, 3H), 6.66 (dd, J = 7. 6,2.4Hz,1H),6.45(d,J=2.4Hz,1H),4.01(s,3H),3.13(s,2H),1.27(s,6H).ESI-MS:m / z=403[M+H] + .

[0667] Example 84: (R)-2-(Difluoromethoxy)-6-methoxy-4-(5-(morpholine-2-methoxy)pyrazolo[1,5-a]pyridin-3-yl)benzonitrile (A95)

[0668]

[0669] Synthesis steps: Referring to step 3 of Example 34, intermediate 18-3 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound 95. 1 H NMR (400MHz, DMSO-d6) δ8.67(d,J=7.6Hz,1H),8.50(s,1H),7.58(t,J=72.8Hz,1H),7.27(dd,J=8.6,1.9Hz,2H),7.20(d,J=1.1Hz,1H),6.77(dd,J=7.6 ,2.6Hz,1H),4.15(d,J=5.0Hz,2H),4.05(s,3H),3.80(dd,J=11.7,2.1Hz,1 H),3.53(td,J=11.1,3.0Hz,2H),2.98-2.56(m,4H).ESI-MS:m / z=431[M+H] + .

[0670] Example 85: (R)-2-(Difluoromethoxy)-6-methoxy-4-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A96)

[0671]

[0672] Synthesis Step 1: Synthesis of Intermediate 96-1

[0673] Synthesis steps: Referring to step 2 of Example 5, intermediate 23-2 was used instead of intermediate 5-2 to obtain intermediate 96-1, ESI-MS: m / z=334 [M+H] + .

[0674] Synthesis step 2: Synthesis of (R)-2-(difluoromethoxy)-6-methoxy-4-(7-(morpholine-2-methoxy)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A96)

[0675] Synthesis steps Referring to step 3 of Example 34, intermediate 96-1 was used instead of intermediate 1-4, and intermediate 93-6 was used instead of intermediate 34-3 to obtain compound A96, ESI-MS: m / z=431 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.59(d,J=7.6Hz,1H),7.84(s,1H),7.51(t,J=72.6Hz,1H),7.25(d,J=1.2Hz,1H),7.17(s,1H),7.04(d,J=2.6Hz,1 H),6.71(dd,J=7.6,2.5Hz,1H),4.20-3.87(m,6H),3.80-3.72(m,2H),2.96-2.87(m,1H),2.80-2.61(m,2H),2.55(dd,J=12.2,10.4Hz,1H).

[0676] Example 86: (R)-2-(Difluoromethoxy)-6-methoxy-4-(7-((pyrrole-3-methylene)amino)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A97)

[0677]

[0678] Synthesis Step 1: Synthesis of Intermediate 97-1

[0679] Synthesis steps: Referring to step 2 of Example 1, intermediate (R)-1-Boc-3-aminomethylpyrrolidine was used to replace 1-tert-butyloxycarbonyl-4-aminomethylpiperidine to obtain intermediate 97-1, ESI-MS: m / z=317 [M+H] + .

[0680] Synthesis step 2: Synthesis of (R)-2-(difluoromethoxy)-6-methoxy-4-(7-((pyrrole-3-methylene)amino)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A97)

[0681] Synthesis steps Referring to step 3 of Example 34, intermediate 97-1 was used instead of intermediate 1-4, and intermediate 93-6 was used instead of intermediate 34-3 to obtain compound A97, ESI-MS: m / z=414 [M+H] + .

[0682] Example 87: (R)-2-(Difluoromethoxy)-6-methoxy-4-(7-((morpholine-2-methylene)amino)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A98)

[0683]

[0684] Synthesis Step 1: Synthesis of Intermediate 98-1

[0685] Synthesis steps: Referring to step 2 of Example 1, intermediate (R)-1-Boc-3-aminomethylpyrrolidine was used to replace 1-tert-butyloxycarbonyl-4-aminomethylpiperidine to obtain intermediate 98-1, ESI-MS: m / z=333 [M+H] + .

[0686] Synthesis step 2: Synthesis of (R)-2-(difluoromethoxy)-6-methoxy-4-(7-((morpholine-2-methylene)amino)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A98)

[0687] Synthesis steps Referring to step 3 of Example 34, intermediate 98-1 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A98, ESI-MS: m / z=430 [M+H] + .

[0688] Synthesis step 3: Synthesis of (R)-2-(difluoromethoxy)-6-methoxy-4-(7-((morpholine-2-methylene)amino)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A98)

[0689] Synthesis steps Referring to step 3 of Example 34, intermediate 98-1 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A98, ESI-MS: m / z=430 [M+H] + .

[0690] Example 88: 2-(Difluoromethoxy)-4-(5-(2-hydroxy-2-methylpropoxy)pyrazolo[1,5-a]pyridin-3-yl)-6-methoxybenzonitrile (A99)

[0691]

[0692] Synthesis Step 1: Synthesis of Intermediate 99-1

[0693] Synthesis steps: Referring to step 2 of Example 1, 1-bromo-2-methyl-2-propanol was used instead of 1-tert-butyloxycarbonyl-4-aminomethylpiperidine to obtain intermediate 99-1, ESI-MS: m / z=207 [M+H] + .

[0694] Synthesis step 2: Synthesis of 2-(difluoromethoxy)-4-(5-(2-hydroxy-2-methylpropoxy)pyrazolo[1,5-a]pyridin-3-yl)-6-methoxybenzonitrile (A99)

[0695] Synthesis steps Referring to step 3 of Example 34, intermediate 99-1 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A99, ESI-MS: m / z=404 [M+H] + .

[0696] Example 89: 2-(Difluoromethoxy)-4-(5-(((1s,4s)-4-hydroxycyclohexyl)oxy)pyrazolo[1,5-a]pyridin-3-yl)-6-methoxybenzonitrile (A100)

[0697]

[0698] Synthesis step 1: Synthesis of intermediate 100-2

[0699] Synthesis procedure: Referring to step 1 of Example 10, trans-1,4-cyclohexanediol was used instead of tert-butyl-4-(2-hydroxyethyl)piperazine-1-carboxylate to obtain intermediate 100-2, which was directly used in the next step.

[0700] Synthesis Step 2: Synthesis of Intermediate 100-3

[0701] Synthesis steps: Refer to step 2 of Example 5, replace 5-2 with intermediate 12-5, and replace 4-(2-bromoethyl)morpholine hydrobromide with intermediate 100-2 to obtain intermediate 100-3, yield: 41%; ESI-MS: m / z=233[M+1] + .

[0702] Synthesis step 3: Preparation of 2-(difluoromethoxy)-4-(5-(((1s,4s)-4-hydroxycyclohexyl)oxy)pyrazolo[1,5-a]pyridin-3-yl)-6-methoxybenzonitrile (A100)

[0703] Synthesis steps: Referring to step 3 of Example 34, intermediate 100-3 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A100, ESI-MS: m / z=430 [M+H] + .

[0704] Example 90: 2-(Difluoromethoxy)-4-(5-(2-(dimethylamino)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-6-methoxybenzonitrile (A101)

[0705]

[0706] Synthesis steps Referring to step 3 of Example 34, intermediate 28-2 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound 101. 1 H NMR (400MHz, DMSO-d6) δ8.67(d,J=7.6Hz,1H),8.51(s,1H),7.58(t,J=72.6Hz,1H),7.32-7.25(m,4H),6.75(dd, J=7.6,2.5Hz,1H),4.26(t,J=5.7Hz,2H),4.06(s,3H),2.72(t,J=5.7Hz,2H),2.25(s,6H).ESI-MS:m / z=403[M+H] + .

[0707] Example 91: 2-(Difluoromethoxy)-6-methoxy-4-(5-(2-morpholinylethoxy)pyrazolo[1,5-a]pyridin-3-yl)benzonitrile (A102)

[0708]

[0709] Synthesis steps: Referring to step 3 of Example 34, intermediate 13-1 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound 102, ESI-MS: m / z=445 [M+H] + . 1H NMR (400MHz, Chloroform-d) δ8.35(d,J=7.6Hz,1H),8.09(s,1H),7.06(d,J=1.4Hz,1H),6.98(d,J=2.6Hz,1H),6.91(d,J=1.4Hz,1H),6.68(t,J= 72.0Hz,1H),6.59(dd,J=7.6,2.6Hz,1H),4.18(t,J=5.4Hz,2H),4.01(s,3H),3.75(t,J=4.7Hz,4H),2.88(t,J=5.4Hz,2H),2.61(t,J=4.7Hz,4H).

[0710] Example 92: 2-(Difluoromethoxy)-4-(7-(2-(dimethylamino)ethoxy)imidazo[1,2-a]pyrazol-3-yl)-6-methoxybenzonitrile (A103)

[0711]

[0712] Synthesis Step 1: Synthesis of Intermediate 103-1

[0713] Synthesis steps: Refer to step 2 of Example 5, replace intermediate 5-2 with intermediate 23-2 to obtain intermediate 103-1, ESI-MS: m / z=206 [M+H] + .

[0714] Synthesis step 2: Preparation of 2-(difluoromethoxy)-4-(7-(2-(dimethylamino)ethoxy)imidazo[1,2-a]pyrazol-3-yl)-6-methoxybenzonitrile (A103)

[0715] Synthesis steps Referring to step 3 of Example 34, intermediate 103-1 was used instead of intermediate 1-4, and intermediate 93-6 was used instead of intermediate 34-3 to obtain compound A103, ESI-MS: m / z=403 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.60(d,J=7.6Hz,1H),7.86(s,1H),7.61(t,J=72.6Hz,1H),7.27(d,J=1.2Hz,1H),7.19(d,J=1.1Hz,1 H),7.08(d,J=2.6Hz,1H),6.70(dd,J=7.6,2.6Hz,1H),4.14(t,J=5.6Hz,2H),4.00(s,3H),2.63(t,J=5.6Hz,2H),2.19(s,6H).

[0716] Example 93: (S)-2-(Difluoromethoxy)-6-methoxy-4-(5-(pyrrole-3-methoxy)pyrazolo[1,5-a]pyridin-3-yl)benzonitrile (A104)

[0717]

[0718] Synthesis steps Referring to step 3 of Example 34, intermediate 58-3 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A104, ESI-MS: m / z=415 [M+H] + .

[0719] Example 94: 2-(Difluoromethoxy)-6-methoxy-4-(7-((piperidin-4-ylmethyl)amino)imidazo[1,2-a]pyridin-3-yl)benzonitrile (A105)

[0720]

[0721] Synthesis steps Referring to step 3 of Example 34, intermediate 1-3 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A105, ESI-MS: m / z=428 [M+H] + . 1 H NMR(400MHz, Methanol-d4)δ8.25(d,J=7.6Hz,1H),7.55(s,1H),7.20(t,J=72.6Hz ,1H),7.07(d,J=1.3Hz,1H),6.99(q,J=1.0Hz,1H),6.56(dd,J=7.5,2.4Hz,1H),6. 33(d,J=2.3Hz,1H),4.00(s,3H),3.16(dd,J=12.4,4.1Hz,2H),3.04(d,J=6.4Hz,2 H),2.70(td,J=12.6,2.5Hz,2H),1.89(dt,J=14.6,2.5Hz,3H),1.26-1.24(m,3H).

[0722] Example 95: 2-(Difluoromethoxy)-4-(6-(2-hydroxy-2-methylpropoxy)pyrazolo[1,5-a]pyridin-3-yl)-6-methoxybenzonitrile (A106)

[0723]

[0724] Step 1: Synthesis of Intermediate 106-2

[0725] 2-Bromo-5-fluoropyridine (5.25 g, 1 eq., 30.0 mmol), DIPEA (4.64 g, 1.2 eq., 36.0 mmol), cuprous iodide (2.87 g, 0.5 eq., 15.0 mmol), (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (Pd(dppf)2Cl2) (487 mg, 0.02 eq, 0.6 mmol), triethylsilyl acetylene (5.04 g, 1.2 eq, 3.6 mmol) were dissolved in DMF and stirred under reflux at 80°C overnight under N2 protection. After the reaction was completed, the mixture was extracted three times with ethyl acetate and saturated brine, the organic phases were combined, concentrated under reduced pressure, and separated by column chromatography (PE:EA=10:1) to obtain 5.8 g of an oily substance (106-2). ESI-MS: m / z=236[M+H] + .

[0726] Step 2: Synthesis of Intermediate 106-4

[0727] 2,4,6-Trimethylbenzenesulfonyl chloride (6.5 g, 1 eq., 30 mmol) and tert-butyl N-hydroxycarbamate (4.0 g, 1 eq., 30 mmol) were dissolved in MTBE, then sealed, replaced with N2, and triethylamine (3.1 g, 1.02 eq., 30.6 mmol) was added dropwise at -5°C. After the reaction was completed, suction was filtered, and the filter residue and filter bottle were washed with MTBE. The filtrate was collected and dried at room temperature, and then slurried with petroleum ether to obtain 9.4 g of white solid 106-4. ESI-MS: m / z=316[M+H] + .

[0728] Step 3: Synthesis of Intermediate 106-5

[0729] The intermediate 106-4 (9.4 g, 1 eq., 29.8 mmol) was placed in a round-bottom flask, and then 30 mL of trifluoroacetic acid was added and stirred at -5°C for 2 hours. After the reaction was completed, water was added and the solid was collected by suction filtration (note: the solid needs to be kept moist). The solid was transferred to a plastic measuring cup with a plastic key, and dichloromethane was added. The organic phase (106-5) was separated and used for the next step. ESI-MS: m / z = 216 [M + H] + .

[0730] Step 4: Synthesis of Intermediate 106-6

[0731] The intermediate 106-5 obtained in the previous step (in DCM) was placed in a round-bottom flask, and then the intermediate 106-2 (5.4 g, 0.77 eq., 23.0 mmol) was added and stirred overnight at room temperature. After the reaction was completed, the reaction solution was distilled under reduced pressure to remove the solvent, and then washed with petroleum ether several times to obtain an oil (intermediate 106-6), which was directly used in the next step. ESI-MS: m / z=252[M+H] + .

[0732] Step 5: Synthesis of Intermediate 106-7

[0733] The intermediate 106-6 obtained in the previous step was dissolved in DMF, and then silver carbonate (380 mg, 0.06 eq., 1.38 mmol) was added and stirred at room temperature overnight. After the reaction was completed, the organic phases were combined after extraction with EA three times, and the solvent was removed by vacuum distillation. Column chromatography (PE:EA=10:1) was used to obtain 1.25 g of brown oil (106-7). ESI-MS: m / z=137[M+H] + .

[0734] Step 6: Synthesis of Intermediate 106-8

[0735] 2-Methyl-1,2-propanediol (216 mg, 2.4 mmol) was dissolved in anhydrous DMF, sodium hydrogen sulfide (96 mg, 2.4 mmol) was added under ice bath, and stirred at room temperature for 0.5 h; then intermediate 106-7 (272 mg, 2.0 mmol) was added, and the temperature was raised to 70°C for overnight reaction. After the reaction was completed, the product was extracted three times with ethyl acetate and saturated brine, the organic phases were combined, concentrated under reduced pressure, and purified by column chromatography to obtain the product intermediate 106-8 (50 mg). ESI-MS: m / z=207[M+H]+.

[0736] Step 7: Synthesis of 2-(difluoromethoxy)-4-(6-(2-hydroxy-2-methylpropoxy)pyrazolo[1,5-a]pyridin-3-yl)-6-methoxybenzonitrile (A106)

[0737] Synthesis steps Referring to step 3 of Example 34, intermediate 106-8 was used to replace intermediate 1-4, and intermediate 93-6 was used to replace intermediate 34-3 to obtain compound A106, ESI-MS: m / z=404 [M+H] +. Chloroform-d)δ7.87(d,J=2.4Hz,1H),7.60(d,J=9.6Hz,1H),7.24(d,J=1.2Hz,1H),7.21(d,J=1.3Hz,1H), 7.17(d,J=9.6Hz,1H),6.69(t,J=71.9Hz,1H),6.60(d,J=2.4Hz,1H),3.95(s,3H),3.76(s,2H),1.17(s,6H).

[0738] Example 96: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(morpholin-2-ylmethoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)methanesulfonamide (A107)

[0739]

[0740] Synthesis steps Referring to step 3 of Example 34, intermediate 77-1 was used instead of intermediate 1-4, and intermediate 78-1 was used instead of intermediate 34-3 to obtain compound A107, ESI-MS: m / z=516 [M+H] + . 1 H NMR(400MHz,Chloroform-d)δ8.42(d,J=2.4Hz,1H),8.20(d,J=7.6Hz,1H),7.89-7.86(m,1H),7.83(s,1H),7.62(s,1H),7.50(s,1H),7.32(d dd,J=10.4,7.9,2.4Hz,1H),7.03(s,1H),6.60(dd,J=7.6,2.5Hz,1H),4.00-3.90(m,4H),3.69-3.63(m,1H),3.09(s,3H),2.95-2.72(m,4H).

[0741] Example 97: (S)-N-(2',4'-difluoro-5-(7-(morpholin-2-ylmethoxy)imidazo[1,2-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)methanesulfonamide (A108)

[0742]

[0743] Synthesis Step 1: Synthesis of Intermediate 108-1

[0744] Synthesis steps: Referring to step 2 of Example 34, methanesulfonic anhydride was used instead of cyclopropanesulfonyl chloride to obtain intermediate 108-1, ESI-MS: m / z=362 [M+H] + .

[0745] Synthesis step 2: Synthesis of (S)-N-(2',4'-difluoro-5-(7-(morpholin-2-ylmethoxy)imidazo[1,2-a]pyridin-3-yl)-[1,1'-biphenyl]-3-yl)methanesulfonamide (A108)

[0746] Synthesis steps Referring to step 3 of Example 34, intermediate 108-1 was used instead of intermediate 1-4, and intermediate 77-1 was used instead of intermediate 34-3 to obtain compound A108, ESI-MS: m / z=515 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.45(d,J=7.6Hz,1H),7.74-7.61(m,2H),7.45(s,3H),7.39(d,J=9.9Hz,2H),7.27-7.19(m,1H),7.08-7.04 (m,1H),6.73(dd,J=7.6,2.5Hz,1H),4.12-4.06(m,2H),4.02(m,1H),3.84(d,J=11.5Hz,2H),3.58(s,2H),3.17(s,2H),3.10(s,3H).

[0747] Example 98: N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)methanesulfonamide (A109)

[0748]

[0749] Synthesis steps Referring to step 3 of Example 34, intermediate 6-3 was used instead of intermediate 1-4, and intermediate 78-1 was used instead of intermediate 34-3 to obtain compound A109, ESI-MS: m / z=530 [M+H] + . 1 H NMR(400MHz,Chloroform-d)δ8.44(s,1H),8.21-8.16(m,1H),8.11(s,1H),7.91(s,1H),7.72(d,J=9.7Hz,1H),7.66(s,1H),7.58(s,1H),7.32( t,J=9.2Hz,1H),7.05(d,J=9.7Hz,1H),4.12(t,J=5.5Hz,2H),3.75(t,J=4.6Hz,4H),3.09(s,3H),2.84(t,J=5.4Hz,2H),2.59(t,J=4.7Hz,4H).

[0750] Example 99: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(pyrrol-3-ylmethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)methanesulfonamide (A110)

[0751]

[0752] Synthesis Step 1: Synthesis of Intermediate 110-2

[0753] Synthesis steps: Refer to step 6 of Example 95, use (S)-1-BOC-3-hydroxymethylpyrrolidine instead of 2-methyl-1,2-propanediol to obtain intermediate 110-2, ESI-MS: m / z=318 [M+H] + .

[0754] Synthesis step 2: Synthesis of (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(pyrrol-3-ylmethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)methanesulfonamide (A110)

[0755] Synthesis steps Referring to step 3 of Example 34, intermediate 110-2 was used to replace intermediate 1-4, and intermediate 78-1 was used to replace intermediate 34-3 to obtain compound A110, ESI-MS: m / z=500 [M+H] + .

[0756] Example 100: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(morpholin-2-ylmethoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)ethylsulfonamide (A111)

[0757]

[0758] Synthesis step 1: Synthesis of intermediate 111-1

[0759] Synthesis steps: Referring to step 2 of Example 34, ethanesulfonic anhydride was used instead of cyclopropanesulfonyl chloride to obtain intermediate 111-1, ESI-MS: m / z=377 [M+H] + .

[0760] Synthesis step 2: Synthesis of (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(7-(morpholin-2-ylmethoxy)imidazo[1,2-a]pyridin-3-yl)phenyl)ethylsulfonamide (A111)

[0761] Synthesis steps Referring to step 3 of Example 34, intermediate 111-1 was used instead of intermediate 1-4, and intermediate 77-1 was used instead of intermediate 34-3 to obtain compound A111, ESI-MS: m / z=530 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ8.70(d,J=2.4Hz,1H),8.41(d,J=7.6Hz,1H),8.13(ddt,J=12.1,8.9,2.0Hz,1H),7 .78(dq,J=8.9,1.6Hz,2H),7.66(s,1H),7.53(t,J=1.9Hz,1H),7.07(d,J=2.6Hz,1H),6.76(dd,J=7.6,2.6 Hz,1H),4.08-4.02(m,2H),3.76(tt,J=10.2,3.9Hz,2H),3.49(td,J=10.5,3.9Hz,1H),3.21(t,J=7.3Hz,2 H), 2.90 (dd, J = 12.1, 2.4Hz, 1H), 2.68 (dd, J = 9.9, 3.0Hz, 2H), 2.55 (d, J = 1.9Hz, 2H), 1.25 (t, J = 7.3Hz, 3H).

[0762] Example 101: N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)-N-methylmethanesulfonamide (A112)

[0763]

[0764] Synthesis step 1: Synthesis of intermediate 112-1

[0765] Intermediate 78-1 (1 mmol) and K2CO3 (1.1 mmol) were placed in a round-bottom flask, and then 5 mL of DMF was added to fully dissolve the raw materials, and CH3I (1.2 mmol) was added dropwise, and stirred at room temperature for 18 h. After the reaction was completed, it was extracted three times with ethyl acetate / water, and the combined organic phases were dried over anhydrous Na2SO4. After the solvent was removed in vacuo, it was purified by column chromatography (PE:EA=3:1) to obtain a light yellow solid product (Intermediate 112-1). Yield: 90%; ESI-MS: m / z=377[M+H] + .

[0766] Synthesis step 2: Synthesis of N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)-N-methylmethanesulfonamide (A112)

[0767] Synthesis steps Referring to step 3 of Example 34, intermediate 6-3 was used instead of intermediate 1-4, and intermediate 112-1 was used instead of intermediate 34-3 to obtain compound A112, ESI-MS: m / z=544 [M+H] + . 1 H NMR(400MHz,Chloroform-d)δ8.46(s,1H),8.10(s,2H),8.04(s,1H),7.80-7.74(m,2H),7.71(s,1H),7.34(t,J=9.2Hz,1H),7.07( d,J=9.7Hz,1H),4.12(t,J=5.5Hz,2H),3.75(t,J=4.7Hz,4H),3.43(s,3H),2.92(s,3H),2.84(t,J=5.6Hz,2H),2.65-2.55(m,4H).

[0768] Example 102: N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)methanesulfonamide (A113)

[0769]

[0770] Synthesis steps Referring to step 3 of Example 34, intermediate 13-1 was used instead of intermediate 1-4, and intermediate 78-1 was used instead of intermediate 34-3 to obtain compound A113, ESI-MS: m / z=530 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.68(s,1H),8.62(d,J=7.6Hz,1H),8.29(s,1H),8.10(t,J=9.5Hz,1H),7.82(s,1H),7.61(s,2H), 7.21(d,J=2.7Hz,1H),6.69(dd,J=7.6,2.5Hz,1H),4.22(t,J=5.7Hz,2H),3.09(s,3H),2.76(t,J=5.7Hz,2H),2.48(m,4H).

[0771] Example 103: (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(6-((4-methylmorpholin-2-yl)methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)-N-methylmethanesulfonamide (A114)

[0772]

[0773] Synthesis Step 1: Synthesis of Intermediate 114-2

[0774] Synthesis steps: Referring to step 6 of Example 95, (S)-4-methyl-2-hydroxymethylmorpholine was used instead of 2-methyl-1,2-propanediol to obtain intermediate 114-2, ESI-MS: m / z=248 [M+H] + .

[0775] Synthesis step 2: Synthesis of (S)-N-(3-(3,5-difluoropyridin-2-yl)-5-(6-((4-methylmorpholin-2-yl)methoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)-N-methylmethanesulfonamide (A114)

[0776] Synthesis steps Referring to step 3 of Example 34, intermediate 114-2 was used to replace intermediate 1-4, and intermediate 112-1 was used to replace intermediate 34-3 to obtain compound A114, ESI-MS: m / z=544 [M+H] + . 1 H NMR(400MHz,Chloroform-d)δ8.47(d,J=2.3Hz,1H),8.11(d,J=3.4Hz,2H),8.04(s,1H) ,7.76(d,J=9.7Hz,2H),7.71(s,1H),7.35(ddd,J=10.4,7.9,2.4Hz,1H),7.10(dd,J=9. 8,2.1Hz,1H),4.06-3.95(m,4H),3.81-3.71(m,1H),3.43(s,3H),2.93(s,3H),2.86(d, J=11.2Hz,1H),2.69(d,J=11.6Hz,1H),2.34(s,3H),2.20(d,J=3.3Hz,1H),2.07(s,1H).

[0777] Example 104: N-(3-(6-(2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-5-(3,5-difluoropyridin-2-yl)phenyl)-N-methylmethanesulfonamide (A115)

[0778]

[0779] Synthesis Step 1: Synthesis of Intermediate 115-2

[0780] In a sealed test tube, a suspension of 2-oxa-7-azaspiro[3,5]nonane oxalic acid (100 mg, 0.46 mmol), potassium carbonate (115 mg, 0.84 mmol) and bromoethanol (30 μL, 0.42 mmol) in dry acetonitrile (1.5 mL) was heated at 90° C. for 16 h. After cooling to room temperature, the suspension was filtered and the filtrate was concentrated in vacuo to give the product as a yellow oil.

[0781] Synthesis Step 2: Synthesis of Intermediate 115-3

[0782] Synthesis steps: Refer to step 6 of Example 95, replace 2-methyl-1,2-propanediol with intermediate 115-2 to obtain intermediate 115-3, ESI-MS: m / z=288 [M+H] + .

[0783] Synthesis step 3: N-(3-(6-(2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)-5-(3,5-difluoropyridin-2-yl)phenyl)-N-methylmethanesulfonamide (A115)

[0784] Synthesis steps Referring to step 3 of Example 34, intermediate 115-3 was used to replace intermediate 1-4, and intermediate 112-1 was used to replace intermediate 34-3 to obtain compound A115, ESI-MS: m / z=584 [M+H] + .

[0785] Example 105: N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(2-(2-methyl-2,7-diazaspiro[3.5]nonan-7-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)-N-methylmethanesulfonamide (A116)

[0786]

[0787] Synthesis Step 1: Synthesis of Intermediate 116-2

[0788] In a sealed test tube, a suspension of 2-methyl-2,7-diazaspiro[3.5]nonane (65 mg, 0.46 mmol), potassium carbonate (115 mg, 0.84 mmol) and bromoethanol (30 μL, 0.42 mmol) in dry acetonitrile (1.5 mL) was heated at 90° C. for 16 h. After cooling to room temperature, the suspension was filtered and the filtrate was concentrated in vacuo to give the product as a yellow oil.

[0789] Synthesis Step 2: Synthesis of Intermediate 116-3

[0790] Synthesis steps: Refer to step 6 of Example 95, use intermediate 116-2 instead of 2-methyl-1,2-propanediol to obtain intermediate 116-3, ESI-MS: m / z=288 [M+H] + .

[0791] Synthesis step 3: N-(3-(3,5-difluoropyridin-2-yl)-5-(6-(2-(2-methyl-2,7-diazaspiro[3.5]nonan-7-yl)ethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)-N-methylmethanesulfonamide (A116)

[0792] Synthesis steps Referring to step 3 of Example 34, intermediate 116-3 was used to replace intermediate 1-4, and intermediate 112-1 was used to replace intermediate 34-3 to obtain compound A116, ESI-MS: m / z=597 [M+H] + .

[0793] Example 106: (3-(3,5-difluoropyridin-2-yl)-5-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)(imino)(methyl)-λ 6 -Sulfone (A117)

[0794]

[0795] Synthesis step 1: Synthesis of intermediate 117-2:

[0796] 1,3,5-Tribromobenzene (3.12 g, 10.0 mmol) and sodium thiomethoxide (3.85 g, 11.0 mmol) were dissolved in DMF and heated to 80°C for 20 h. After the reaction was completed, the mixture was extracted three times with petroleum ether and saturated brine, the organic phases were combined, concentrated under reduced pressure, and purified by column chromatography to obtain the product 117-2 (2.0 g).

[0797] Synthesis step 2: Synthesis of intermediate 117-3:

[0798] The intermediate 117-2 (560 mg, 2.0 mmol) and triisopropyl borate (376 mg, 2.0 mmol) were dissolved in anhydrous tetrahydrofuran, and n-butyl lithium (0.8 mL, 2.0 mmol, 2 M) was added dropwise at -78 °C and the reaction was continued for 1 h. After the reaction was completed, saturated ammonium chloride solution was used to quench and the mixture was restored to room temperature. The mixture was extracted three times with ethyl acetate and saturated brine, and the organic phases were combined, concentrated under reduced pressure, and purified by column chromatography to obtain the product 117-3 (470 mg). ESI-MS: m / z=247[M+H]+

[0799] Synthesis step 3: Synthesis of intermediate 117-5:

[0800] The intermediate 117-3 (348 mg, 1.4 mmol), 2-bromo-3,5-difluoropyridine (273 mg, 1.4 mmol), Pd(PPh3)4 (81 mg, 0.07 mmol), potassium carbonate (389 mg, 1.2 mmol) were dissolved in a mixed solution of 1,4-dioxane and water (1,4-dioxane: water = 8:1), replaced with nitrogen protection, and heated to 90 ° C for 8 hours. After the reaction was completed, it was extracted three times with ethyl acetate and saturated brine, the organic phases were combined, concentrated under reduced pressure, and purified by column chromatography to obtain the product 117-5 (180 mg). ESI-MS: m / z = 316 [M + H] +

[0801] Synthesis Step 4: Synthesis of Intermediate 117-6:

[0802] The intermediate 117-5 (130 mg, 0.41 mmol), ammonium carbonate (83 mg, 0.86 mmol) and iodophenyl diacetic acid (200 mg, 0.64 mmol) were dissolved in methanol and reacted at room temperature for 5 h. After the reaction was completed, the mixture was directly concentrated under reduced pressure and purified by column chromatography to obtain the product 117-6 (63 mg). ESI-MS: m / z=347[M+H]+

[0803] Synthesis step 5: (3-(3,5-difluoropyridin-2-yl)-5-(6-(2-morpholinethoxy)pyrazolo[1,5-a]pyridin-3-yl)phenyl)(imino)(methyl)-λ 6 -Synthesis of sulfone (A117):

[0804] Intermediate 117-6 (50 mg, 0.14 mmol), intermediate 13-1 (36 mg, 0.14 mmol), Pd(dppf)2Cl2 (10 mg, 0.014 mmol), potassium acetate (28 mg, 0.28 mmol) were dissolved in DMF, replaced with nitrogen protection, and heated to 110°C for 18 h. After the reaction was completed, it was extracted three times with ethyl acetate and saturated brine, the organic phases were combined, concentrated under reduced pressure, and purified by column chromatography to obtain product A117 (10 mg). ESI-MS: m / z=514[M+H]+.

[0805] Example 107: 4-(2-((3-(3-(3,5-difluoropyridin-2-yl)-5-(methylsulfonyl)phenyl)pyrazolo[1,5-a]pyridin-6-yl)oxy)ethyl)morpholine (A118)

[0806]

[0807] Synthesis step 1: Synthesis of intermediate 118-1

[0808] The intermediate 117-5 (800 mg, 2.54 mmol) was dissolved in 30 ml DCM and 10 ml H2O. At 0°C, m-CPBA (1.55 g, 7.62 mmol) was added in batches to warm the reaction to room temperature. The mixture was stirred for 4 h. The reaction solution was poured into 30 ml 0°C ice water. Sodium thiosulfate was added to neutralize the unreacted peroxide (PH=8). The reaction solution was extracted with DCM (3x10 ml). The organic phases were combined, washed with saturated sodium chloride aqueous solution (4x5 ml), and then dried over anhydrous sodium sulfate. The organic phase was mixed with 1 g of silica gel and purified by column chromatography (PE:EA=0-35%, 35 min) to obtain 534 mg of an off-white solid. ESI-MS: m / z=348[M+H]+; the yield was 60.61%.

[0809] Synthesis step 2: Synthesis of 4-(2-((3-(3-(3,5-difluoropyridin-2-yl)-5-(methylsulfonyl)phenyl)pyrazolo[1,5-a]pyridin-6-yl)oxy)ethyl)morpholine (A118)

[0810] Intermediate 118-1 (253 mg, 0.73 mmol), intermediate 13-1 (150 mg, 0.61 mmol), Pd(dppf)Cl2 (89 mg, 0.12 mmol) and KOAc (178 mg, 1.82 mmol) were dissolved in 10 ml DMF, N2 was replaced, and the reaction was carried out at 110 ° C overnight. The reaction was cooled to room temperature and poured into 10 ml water. The mixture was extracted with EA (2x10 ml), the organic phases were combined, washed with saturated brine (2x5 ml), and then dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. Through a rapid purification device (5 mmol NH4HCO3 / H2O: ACN = 0-55%, 45 min), 36.4 mg of white solid was obtained, ESI-MS: m / z = 515 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ8.69 (d, J=2.4 Hz, 1H), 8.53-8.48 (m, 1H), 8.42 (s, 1H), 8.37 (s, 1H), 8.26-8.09 (m, 3H), 7.87 (d, J=9.7 Hz, 1H), 7.22 (dd, J=9.7, 2.1 Hz, 1H), 4.14 (t, J=5.6 Hz, 2H), 3.55 (t, J=4.6 Hz, 4H), 3.31 (s, 3H), 2.69 (t, J=5.6 Hz, 2H). 2.50-2.44 (m, 4H); the yield was 11.67%.

[0811] Example 108 Biological Activity Experiment

[0812] 1. In vitro SIKs kinase activity inhibition experiment

[0813] ADP-Glo TM Kinase assay is a luminescent technique analysis of the ADP formed by the kinase reaction. In this particular study, the kinase reaction is composed of the phosphorylation of SIK1 (NCBI Reference Sequence: NP_775490.2), SIK2 (NCBI Reference Sequence: NP_001353615.1) or SIK3 (NCBI Reference Sequence: NP_001353615.1) by AMARA peptide substrate (manufacturer: Sangon Biotech, catalog number: P23855). In the second step, the kinase reaction is terminated and all remaining ATP is exhausted. In the final step, ADP is converted into ATP and the newly synthesized ATP is measured by using luciferase / luciferin reaction. The light generated is measured using the Envision plate reader, and the luminescent signal obtained is positively correlated with the kinase activity.

[0814] Detection Kit: ADP-Glo TM Kinase Assay (Manufacturer: Promega, catalog number: V9102)

[0815] Reaction system:

[0816] 1. Prepare kinase reaction buffer: 25mM Tris-HCl (pH 7.5), 0.01% TritonX-100, 0.5mM EGTA, 2.5mM DTT, 5mM MgCl2

[0817] 2. Compound preparation: All compounds were dissolved and diluted in 100% DMSO.

[0818] 3. Take 5 μL enzyme and 39.5 μL kinase reaction buffer in a 96-well plate, add 0.5 μL DMSO / positive drug / compound, and use them as DMSO control group, positive drug control group, and compound group respectively.

[0819] 4. Enzyme inactivation treatment: Take SIK1 / 2 / 3 enzyme and heat it in a 95℃ metal bath for 15 minutes. Take 5μL of inactivated enzyme and 39.5μL of kinase reaction buffer in a 96-well plate, and add 0.5μL of DMSO as the enzyme inactivation group.

[0820] 5. Take 4 μL of substrate AMARA (final concentration of 45 μM) and 1 μL of ATP (final concentration of 5 μM), mix them, and add them to the 96-well plates of each group mentioned above.

[0821] 6. Centrifuge the 96-well plate at 1000 rpm for 10 seconds and incubate at room temperature for 1.5 hours.

[0822] 7. Take 5 μL of the reaction solution into a 384-well white plate, add 5 μL of ADP-Glo ​​reagent to stop the reaction and consume the unreacted ATP, centrifuge the 384-well plate at 1000 rpm for 10 seconds, and incubate at room temperature for 40 minutes.

[0823] 8. Add 10 μL of kinase assay reagent to the 384-well plate to convert ADP into ATP, centrifuge the 384-well plate at 1000 rpm for 10 seconds, and incubate at room temperature for 30 minutes.

[0824] 9. Detection of luminescence on a Tecan (SPARK) microplate reader

[0825] Data processing:

[0826] The inhibition rate was calculated as follows:

[0827] Inhibition rate (%) = [1-(RLU sample-RLU enzyme inactivation group) / (RLU DMSO group-RLU enzyme inactivation group)] × 100%

[0828] Data Analysis:

[0829] The log value of the concentration was used as the X-axis and the percentage inhibition rate was used as the Y-axis. The [Inhibitor] vs. normalized response--Variable slope of the analysis software GraphPad Prism9 was used to fit the dose-effect curve to obtain the IC50 value of each compound on the enzyme activity.

[0830] Table 1: Inhibitory activity of compounds against SIKs kinases

[0831]

[0832]

[0833]

[0834]

[0835]

[0836] Note: ND means not detected

[0837] Conclusion: The compounds of the present invention have excellent in vitro inhibitory activity against kinases SIK1 / 2 / 3. In particular, compounds A34-A81 and A107-A118 have excellent inhibitory effects on SIK2 / 3, which are significantly better than comparative compound 1; they also have inhibitory activity against SIK1 and are better than comparative compound 1. Compounds A93-A106 have significantly better inhibitory activity against SIK1 than comparative compound 2.

[0838] 2. LPS-induced acute inflammation in mice

[0839] Lipopolysaccharide (LPS) is a component of the outer membrane of Gram-negative bacteria. The administration of LPS can induce systemic inflammation and cause a strong immune response in animals, which is mainly manifested as uncontrolled activation of inflammatory cells such as neutrophils and monocytes / macrophages, and the release of a large number of inflammatory factors, such as TNF-α.

[0840] Experimental materials: Lipopolysaccharide from Escherichia coli 055:B5 (LPS, manufacturer: Sigma-Aldrich, catalog number: L2880), Mouse TNF alpha Uncoated ELISA Kit (manufacturer: ThermoFishe, catalog number: 88-7324-88), Mouse IL-10 Uncoated ELISA Kit (manufacturer: ThermoFishe, catalog number: 88-7105-88).

[0841] Experimental animals: C57BL / 6N mice (male, SPF grade, 6-8 weeks old, Beijing Weitonglihua Experimental Animal Technology Co., Ltd.)

[0842] Experimental operation:

[0843] The experimental group mice were orally gavaged with each test compound, and the control group and the modeling group were gavaged with an equal amount of solvent; at the Tmax of the test drug, LPS (20 mg / kg, 5 mL / kg) was injected intraperitoneally into the C57BL / 6N mice in the modeling group and the experimental group, and the control group mice were injected with sterile PBS.

[0844] Mice were sacrificed 90 minutes after LPS injection, and blood was collected in 1.5 mL EP tubes and centrifuged at low temperature to obtain serum, which was then subjected to cytokine analysis.

[0845] Cytokine detection:

[0846] The steps for IL-10 and TNF-α are the same and are carried out synchronously.

[0847] 1. Prepare the coating reagent with diluted 1× PBS, that is, add the capture antibody to PBS, add 100 μL of coating reagent to each well of the ELISA plate, and incubate at 4°C overnight.

[0848] 2. Prepare the ELISA plate in advance and bring it to room temperature. Pour out the coating reagent, add washing solution to each well, incubate for 1 minute, discard the washing solution, repeat 3 times, and completely pour out the residual liquid for the last time.

[0849] 3. Add 200 μL 1×ELISA Diluent to each well and block at room temperature for 1 hour.

[0850] 4. Wash at least once with washing solution, prepare standards and samples in advance, and dilute the standards in gradients according to the requirements of the kit; add 100μL 1×ELISA Diluent to the blank wells, and perform duplicate detection on both the standards and blank wells; dilute the samples with 1×ELISA Diluent, the dilution ratio of IL-10 is 1:4, and the dilution ratio of TNF is 1:2, and add 100μL to each well. After sealing with sealing film, incubate at room temperature for 2 hours.

[0851] 5. Dilute the detection antibody in advance at a dilution ratio of 1:250. Repeat step 2 for 3-5 times, add 100 μL of detection antibody to each well, seal the plate and incubate at room temperature for 1 hour.

[0852] 6. Dilute horseradish peroxidase in advance at a dilution ratio of 1:100. Repeat step 2 for 3-5 times, add 100 μL horseradish peroxidase to each well, seal the plate and incubate at room temperature for 30 minutes.

[0853] 7. Repeat the washing in step 2 5-7 times, add 100 μL 1× TMB to each well, and incubate at room temperature for 15 minutes.

[0854] 8. After color development, add 100 μL of stop solution to each well, and immediately read the OD 450nm on a microplate reader and process the data.

[0855] The calibration data were performed in the same experimental method.

[0856] Data processing:

[0857] The standard curve fitting formula was calculated using ELISACalc to obtain the cytokine concentration (CONC) in each well.

[0858] The calculation is as follows:

[0859] Relative expression of TNFα=[(CONC experimental group×dilution factor) / (CONC modeling group×dilution factor)]×100%

[0860] Relative expression of IL-10 = [(CONC experimental group × dilution factor) / (CONC modeling group × dilution factor)] × 100%

[0861] Table 2: Efficacy of compounds in LPS-induced acute inflammation model in mice

[0862]

[0863] Conclusion: The compounds in the table can reduce TNFα and increase IL-10 in LPS-induced acute inflammation in mice, showing certain anti-inflammatory activity in animals.

[0864] 3. Pharmacokinetic testing

[0865] The experiment was completed at Hangzhou Lead Pharmaceutical Technology Co., Ltd.

[0866] Experimental materials: SD rats (male, 7-9 weeks old, Shanghai Slake)

[0867] Experimental procedure: The clear solution obtained after dissolving the test compound was administered orally to male SD rats (overnight fasting, 7-9 weeks old). After administration of the test compound or control compound, blood was collected from the eye sockets at 0.25, 0.50, 1.00, 2.00, 4.00, 6.00, 8.00 and 24.00 hours, and plasma was obtained after centrifugation. The blood drug concentration was determined by LC-MS / MS, and the relevant pharmacokinetic parameters were calculated by the non-compartmental linear logarithmic trapezoidal method using WinNonLin 8.0 pharmacokinetic statistical software.

[0868] Preparation of standard curve and quality control samples: The mixed stock solution of the compound was diluted with 50% methanol water to prepare standard working solutions containing 20, 40, 100, 200, 400, 1000, 2000, 4000, and 10000 ng / mL of each compound, and quality control working solutions of 60, 600, and 8000 ng / mL. 2.50 μL of standard curve working solution and quality control working solution were added to 47.5 μL of blank rat plasma to prepare standard curves containing 1.00, 2.00, 5.00, 10.00, 20.00, 50.00, 100.00, 200.00, 500.00 ng / mL of each compound and quality control samples with concentrations of 3.00, 30.00 and 400.00 ng / mL, and 200 μL of acetonitrile (containing internal standard verapamil 2 ng / mL) was added respectively. After vortexing for 3 min, the mixture was centrifuged at 20000 rcf at 4 °C for 10 min, and the supernatant was taken for LC-MS / MS analysis.

[0869] Preparation and processing of unknown samples: Take 50 μL of plasma sample, add 200 μL of acetonitrile (containing internal standard verapamil 2 ng / mL), vortex for 3 minutes, centrifuge at 20000 rcf, 4°C for 10 minutes, and take the supernatant for LC-MS / MS analysis.

[0870] The unknown samples were diluted 10 times and prepared: 45 μL of blank plasma was taken, 5 μL of plasma sample was added, and 200 μL of acetonitrile (containing internal standard verapamil 2 ng / mL) was added. After vortexing for 3 min, centrifuged at 20000 rcf and 4 °C for 10 min, and the supernatant was taken for LC-MS / MS analysis.

[0871] Table 3: Pharmacokinetic properties of compounds in rats

[0872]

[0873] Conclusion: The compound of the present invention has good oral pharmacokinetic properties. Compared with the comparative compound 2, it has a longer half-life and a higher oral exposure (area under the AUC curve).

[0874] IV. Experimental study on the inhibitory activity of compounds on RIPK2 kinase

[0875] Experimental materials: ADP-Glo ​​Kinase Assay (Promega, V9101); RIPK2 (ICE, S2303T-H17G); WEHI-345 (MCE, HY-18937).

[0876] Experimental procedure: Starting from the highest concentration of 10000nM, prepare the test compound into a 10-point dose-corresponding serial dilution with DMSO in a dilution step of 1 / 4 (the highest concentration of WEHI-345 is 1000nM, and the dilution multiple is 1 / 3), and transfer 0.1μL of the dilution to the 384-well assay plate. Each concentration of the test compound has two replicate wells. Add 5μL of enzyme working solution to each well of the 384-well assay plate, centrifuge at 1000rpm for 1min, and then incubate at 25℃ for 10min. Add 5μL of ATP working solution to each well to initiate the reaction and incubate at 25℃ for 180min. Add 5μL of ADP-Glo ​​reagent, centrifuge at 1000rpm for 1min, and incubate at 25℃ for 40min. Add 10μL of detection reagent, centrifuge at 1000rpm for 1min, and incubate at 25℃ for 40min. Detect the luminescent signal in an ELISA reader.

[0877] Data processing: Inhibition rate (%) = 100*(ave High control-cpd well) / (ave High control-ave low control), where High control group: no compound added, low control group: no compound and enzyme added. The log value of the concentration was used as the X-axis, and the percentage inhibition rate was used as the Y-axis. The IC value of the test compound of the nonlinear regression equation was calculated using XLfit 5.5.0. 50 Fit the line to obtain the IC value of each compound on the enzyme activity50 value.

[0878] The experimental results are shown in the table below.

[0879] Table 4: Inhibitory activity of compounds against RIPK2 kinase

[0880] Compound No. <![CDATA[RIPK2 IC 50 (nM) <!-- 71 -->]]> GLPG3970 17.5 WEHI-345 17.8 A103 1.93

[0881] Conclusion: The inhibitory activity of the representative compounds of the present invention on RIPK2 kinase is significantly better than that of the RIPK2 kinase-positive drug WEHI-345, and better than that of the SIK kinase-positive drug GLPG3970.

[0882] V. Inhibitory effect of compounds on TNF-α secretion in BMDM cells under MDP stimulation

[0883] Experimental materials: MDP (InvivoGen, tlrl-mdp); Mouse TNF-α Precoated ELISA Kit (Dakoway Biotechnology Co., Ltd., 1217202); IFN-γ (PepreTech, 315-05-20UG); WEHI-345 (MCE, HY-18937).

[0884] Experimental operation: Bone marrow cells of 6-8 week old C57BL / 6 mice were isolated and seeded in 24-well plates (culture conditions: DMEM+50ng / mL M-CSF+10% FBS). The cells were cultured to the 6th day, and WEHI-345, GLPG3940, A18 and A26 were treated with concentration gradients of 2000nM, 500nM, 125nM, 31.25nM, 7.81nM and 1.95nM for 30 minutes. Except for the control group, the other groups were treated with IFN-γ (5ng / mL) for 2h, and then stimulated with MDP (10ng / mL). After 24h, the cell supernatant was collected and centrifuged at 2000-3000rpm for 20min. After that, the supernatant was aspirated into a new EP tube for standby use, and the TNF-α content in the supernatant was detected by ELISA kit.

[0885] Data processing: Inhibition rate (%) = 100*(model-cpd) / (model-control), where model group: no compound added, control group: no compound added and MDP. With the concentration value as the X-axis and the percentage inhibition rate as the Y-axis, GraphPad Prism9 was used to fit the dose-effect curve to obtain the IC value of each compound on the inhibitory activity of TNF-α secretion by BMDM cells under MDP stimulation. 50 value.

[0886] The experimental results are shown in the following table and Figure 1.

[0887] Table 5: Inhibitory activity of compounds on TNF-α secretion by BMDM cells stimulated by MDP

[0888] Compound No. <![CDATA[TNF-α inhibition IC 50 (nM)]]> GLPG3970 119.2 WEHI-345 70.85 A103 4.827

[0889] Conclusion: The representative compounds of the present invention have significantly better inhibitory activity on TNF-α secretion of BMDM cells than WEHI-345 and GLPG3970 at the cellular level.

[0890] VI. Inhibition of NF-κB gene expression in THP-1 cells by compounds under MDP stimulation

[0891] Experimental materials: MDP (InvivoGen, tlrl-mdp); THP-1-Dual TM Cells (InvivoGen, thpd-nfis); QUANTI-Blue (InvivoGen, rep-qbs); WEHI-345 (MCE, HY-18937).

[0892] Experimental steps: 1×10 6 Resuspend THP-1-Dual at a concentration of 100 / mL TM Cells were seeded in 96-well plates (culture conditions: RPMI-1640 + 10% FBS + 2mM L-Glu + 25mM HEPES + 5μg / mL Blasticidin + 50μg / mL Zeocin), 100μL of cell solution per well, cultured for 12h. After that, 100μL of culture medium containing gradient concentrations (8000nM, 2666nM, 888nM, 296nM, 98.7nM, 32.9nM, 10.9nM, 3.65nM, 1.21nM, 0.4nM) of WEHI-345, GLPG3970, A18 and A26 was added to each well for 30min. Except for the control group, 10μg / mL of MDP was added to each well and cultured for 24h. Thereafter, 20 μL of the culture supernatant was aspirated and mixed with 180 μL of QUANTi-Blue reagent, incubated at 37° C. for 4 h, and the absorbance was measured at 620 nm.

[0893] Data processing: Inhibition rate (%) = 100*(model-cpd) / (model-control), where model group: no compound added, control group: no compound added and MDP. With concentration value as X-axis and percentage inhibition rate as Y-axis, GraphPad Prism9 was used to fit the dose-effect curve to obtain the IC value of each compound's inhibitory activity on NF-κB gene expression in THP-1 cells stimulated by MDP.50 value.

[0894] The experimental results are shown in the following table and Figure 2 .

[0895] Table 6: Inhibitory activity of compounds on NF-κB gene expression in THP-1 cells stimulated by MDP

[0896] Compound No. <![CDATA[NF-κB gene inhibition IC 50 (nM)]]> WEHI-345 85.69 A103 7.314

[0897] Conclusion: The representative compounds of the present invention have significantly better inhibitory activity on NF-κB gene expression in THP-1 cells than WEHI-345.

[0898] VII. Effect of Compound A103 on Growth and Weight Gain in ICR Mice

[0899] The experiment was completed at Hangzhou Yuhong Pharmaceutical Technology Co., Ltd.

[0900] Experimental materials: ICR mice (male, 3-4 weeks old, Hangzhou Ziyuan).

[0901] Experimental operation: The control group (N=6) was gavaged with solvent once a day; the drug-treated group (N=6) was gavaged with 25 mg / kg compound suspension once a day, and samples were collected after 28 consecutive days of administration. The compound solvent was CMC-Na, and the single administration volume was 0.1 mL / 10 g. The compound was prepared by mixing the solvent in a mortar and grinding it into a suspension for use.

[0902] Experimental detection indicators: monitoring of body weight changes, sampling of mouse leg bones to measure bone length after stable weight gain.

[0903] The results are shown in the table below and Figure 3 .

[0904] Table 7: Effects of compounds on body weight and leg bone length in ICR mice

[0905] Group Initial weight D28 weight Weight gain rate Mouse leg bone length Control group 20.67±0.12 32.00±0.82 1.55±0.04 3.29±0.07 25mg / kg group 20.83±1.20 36.03±0.67 1.73±0.08 3.48±0.12 p-value NS ＜0.0001 0.0001 0.0016

[0906] Conclusion: The representative compounds of the present invention have a significant weight-increasing effect on ICR young mice and promote the growth of leg bones in mice.

[0907] 8. Experimental study on the effect of compound A103 on cisplatin-induced cachexia in mice

[0908] The experiment was completed at Hangzhou Yuhong Pharmaceutical Technology Co., Ltd.

[0909] Experimental materials: C57BL / 6J mice (male, 8-10 weeks old, Hangzhou Ziyuan).

[0910] Modeling method: Cisplatin (2.5 mg / kg, Jiangsu Hausen) was intraperitoneally administered at 9 am on days 1 to 5, and the test molecule was administered 6 hours after the administration of cisplatin on day 5. The compound solvent was CMC-Na.

[0911] Dosage regimen: control group (N=5), vehicle was gavaged once a day starting from the 5th day; model group (N=6), 25 mg / kg of compound suspension was gavaged once a day after modeling was completed; model+GLPG3970 group (N=6), 25 mg / kg of GLPG3970 solution was gavaged once a day after modeling was completed; model+Yilizhi group (N=6), 160 mg / kg of Yilizhi suspension was gavaged once a day after modeling was completed.

[0912] Experimental detection indicators: weight change monitoring.

[0913] The results are shown in the table below and Figure 4 .

[0914] Table 8: Effects of compounds on body weight of mice with cisplatin-induced cachexia

[0915] Group Initial weight D28 weight Weight gain rate Control group 22.86±0.68 24.10±0.50 1.06±0.01 Model Group 24.17±0.51 22.75±1.05 0.96±0.02 Model + Yilizhi Group 22.91±0.41 22.44±0.43 0.98±0.03 Model + GLPG3970 group 22.80±0.70 21.73±0.54 0.96±0.04 Model + Compound Group 22.85±0.43 23.43±0.86 1.03±0.03

[0916] Conclusion: The representative compounds of the present invention have a significant effect on promoting weight recovery in cisplatin-induced cachexia. Their weight gain effect is not only better than the positive drug megestrol acetate (Yilizhi, Qingdao Guohai) for the clinical treatment of cachexia, but also better than the SIK kinase positive drug GLPG3970.

[0917] The structural formulas of the comparative compounds 1 and 2 used in the experimental examples are shown below:

[0918] .

Claims

1. Aromatic fused ring compounds, characterized in that: It has the structure shown in Formula I: or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Structural unit Selected from or Y and W are each independently selected from C or N; V, Z are selected from CH or N; X is selected from N or CR g ; W, Y, and Z are not all N; W, Y, and Z are not all C; L1 is selected from O or NH or CH2; L2 is selected from C 1-3 Alkanes; R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, unsubstituted or methyl-substituted 5-6-membered aromatic heterocyclic group, unsubstituted or methyl-substituted 9-10-membered fused heteroaromatic ring group, 4, 5 or 6-membered heterocyclic group, the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, the 4, 5 or 6-membered heterocyclic group can be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution; R a , R b Each independently selected from H, C 1-3 Alkyl; R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2; m is selected from 1, 2 or 3; R4 is selected from H, halogen, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy, -NHS(O)2R d 、-NR e S(O)2R d , -S(O)2R d ; R d , R e Each independently selected from C 1-3 Alkyl, cyclopropyl; R5 is selected from -C(O)R f 、-S(O)2R f 、-C(O)NHR f ; R f Selected from C 1-3 Alkyl, cyclopropyl, halogen-substituted C 1-3 Alkyl, -CH2OH; R6, R7, R8, R g Each independently selected from H, F, Cl, methyl, methoxy, trifluoromethoxy, difluoromethoxy, trifluoromethyl; R7 and R8 may form Wherein R6, V, and X are as defined above; R 10 and R 11 Each independently selected from H, C 1-3 Alkyl, one or more fluorine-substituted C 1-3 Alkyl, F, 2. The aromatic fused ring compound according to claim 1, characterized in that: The compound has a structure shown in Formula II: or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Y and W are each independently selected from C or N; Z is selected from CH or N; W, Y, and Z are not all N; W, Y, and Z are not all C; L1 is selected from O or NH or CH2; L2 is selected from C 1-3 Alkanes; R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, unsubstituted or methyl-substituted 5-6-membered aromatic heterocyclic group, unsubstituted or methyl-substituted 9-10-membered fused heteroaromatic ring group, 4, 5 or 6-membered heterocyclic group, the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, the 4, 5 or 6-membered heterocyclic group can be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution; R a , R b Each independently selected from H, C 1-3 Alkyl; R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2; m is selected from 1, 2 or 3; R4 is selected from H, F, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy, -NHS(O)2R d ; R5 is selected from -C(O)R f 、-S(O)2R f 、-C(O)NHR f ; R f Selected from C 1-3 Alkyl, cyclopropyl, -CF3, -CH2CF3, -CH2OH; R d Selected from C 1-3 Alkyl, cyclopropyl.

3. The aromatic fused ring compound according to claim 1, characterized in that: The compound has a structure shown in Formula III: or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Y and W are each independently selected from C or N; Z is selected from CH or N; V is selected from CH or N; X is selected from N or CR g ; W, Y, and Z are not all N; W, Y, and Z are not all C; L1 is selected from O or NH or CH2; L2 is selected from C 1-3 Alkanes; R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, unsubstituted or methyl-substituted 5-6-membered aromatic heterocyclic group, unsubstituted or methyl-substituted 9-10-membered fused heteroaromatic ring group, 4, 5 or 6-membered heterocyclic group, the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, the 4, 5 or 6-membered heterocyclic group can be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution; R a , R b Each independently selected from H, C 1-3 Alkyl; R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2; m is selected from 1, 2 or 3; R4 is selected from F, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy, -NHS(O)2R d 、-NR e S(O)2R d 、- -S(O)2R d ; R d , R e Each independently selected from C 1-3 Alkyl, cyclopropyl; R6, R7, R8, R g Each independently selected from H, F, Cl, methyl, methoxy, trifluoromethoxy, difluoromethoxy, trifluoromethyl; R7 and R8 may form wherein R6, V, and X are as defined above.

4. The aromatic fused ring compound according to claim 1, characterized in that: The compound has a structure shown in Formula IV: or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Y and W are each independently selected from C or N; Z is selected from CH or N; W, Y, and Z are not all N; W, Y, and Z are not all C; L1 is selected from O or NH or CH2; L2 is selected from C 1-3 Alkanes; R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, 4, 5 or 6-membered heterocyclic group, wherein the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, and the 4, 5 or 6-membered heterocyclic group may be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution; R a , R b Each independently selected from H, C 1-3 Alkyl; R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2; m is selected from 1, 2 or 3; R4 is selected from F, C 1-3 Alkoxy, halogen substituted C 1-3 Alkoxy; R 10 and R 11 Each independently selected from H, C 1-3 Alkyl, one or more fluorine-substituted C 1-3 Alkyl, F, 5. The aromatic fused ring compound according to claim 4, characterized in that: The compound has a structure as shown in Formula V, Formula VI, Formula VII, Formula VIII, or Formula IX: or a stereoisomer or a pharmaceutically acceptable salt thereof, in: L1 is selected from O or NH or CH2; L2 is selected from C 1-3 Alkanes; R1 is selected from R c substituted 5-6 membered cycloalkyl, 7-12 membered spirocycloalkyl, 7-12 membered spirocycloheteroalkyl, 7-12 membered bridged cycloalkyl, 7-12 membered bridged cycloheteroalkyl, 8-12 membered paracycloalkyl, 8-12 membered paracycloheteroalkyl, -NR a R b , -OH, 4, 5 or 6-membered heterocyclic group, wherein the 4, 5 or 6-membered heterocyclic group contains 1 or 2 N atoms, and the 4, 5 or 6-membered heterocyclic group may be methyl, fluorine, -(CH2) m OH; the 7-12-membered spirocycloalkyl, 7-12-membered spirocycloheteroalkyl, 7-12-membered bridged cycloalkyl, 7-12-membered bridged cycloheteroalkyl, 8-12-membered paracycloalkyl, 8-12-membered paracycloheteroalkyl may be C 1-3 Alkyl, -OH, -NH2, -NHCH3, -N(CH3)2, halogen, halogen-substituted C 1-3 Alkyl, C 1-3 Alkoxy substitution; R a , R b Each independently selected from H, C 1-3 Alkyl; R c Selected from -OH, -NH2, -NHCH3, -N(CH3)2; m is selected from 1, 2 or 3; R 10 and R 11 Each independently selected from H, C 1-3 Alkyl, one or more fluorine-substituted C 1-3 Alkyl, F, 6. The compound of formula I according to claim 1, characterized in that The unit is selected from the following structures:

7. The compound according to claim 1, characterized in that The unit is selected from the following structures: R6, R7, R8, R g Each independently selected from H, F, Cl, methyl, methoxy, trifluoromethoxy, difluoromethoxy, trifluoromethyl; R7 and R8 may form 8. The compound according to claim 1, or a stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from:

9. A pharmaceutical composition of a pharmaceutically acceptable carrier, excipient, auxiliary material or solvent, characterized in that: The pharmaceutical composition comprises one or more of the compound according to any one of claims 1 to 9, its isomers, and pharmaceutically acceptable salts thereof; the pharmaceutical composition is in the form of tablets, capsules, pills, granules, powders, suppositories, injections, solutions, suspensions, ointments, patches, lotions, drops, wipes, or sprays.

10. Use of the compound according to any one of claims 1 to 8, its isomer, its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 9 in the preparation of a drug for preventing or treating a disease caused by abnormal activation of SIK kinase or a disease accompanied by weight loss, wherein the disease caused by abnormal activation of SIK kinase is selected from at least one of cancer, immune inflammation, diabetes and central nervous system diseases; wherein, The cancer is selected from at least one of ovarian cancer, breast cancer, prostate cancer, glioma, and diffuse large B-cell lymphoma; the immune inflammation is selected from at least one of arthritis, colitis, psoriasis, atopic dermatitis, psoriatic arthritis, systemic lupus erythematosus, osteoarthritis, complications caused by organ transplantation, autoimmune thyroid disease, Sjögren's syndrome, vitiligo, autoimmune kidney damage, autoimmune liver damage, chronic obstructive pulmonary disease, multiple sclerosis, amyotrophic lateral sclerosis, giant cell arteritis, Takayasu arteritis, and sepsis; the central nervous system disease is selected from at least one of depression, insomnia, and epilepsy; the disease accompanied by weight loss is selected from at least one of cachexia, growth retardation, emaciation, and skeletal muscle muscular atrophy; the cachexia is selected from at least one of cachexia associated with cancer, acquired immunodeficiency syndrome, heart failure, and / or chronic kidney disease.

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