N 2 - Substituted bicyclo-2-aminopyrimidine derivatives, processes for their preparation and medical use

Abstract

The application discloses a compound shown in formula (I), a preparation method and medical application of the compound, and optical isomers and pharmaceutically acceptable salts of the compound. The compound has FLT3 and CHK1 inhibiting activity, has proliferation inhibiting activity on multiple leukemia cell strains, is effective on multiple AML mutations such as internal tandem duplication mutations of a near membrane domain and D835 point mutations of an activation loop in a kinase domain, can overcome drug resistance caused by point mutations in clinic, and can be applied to preparation of an antitumor drug.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, and more particularly to an N 2 - Replacement of bicyclic-2-aminopyrimidine derivatives, optical isomers, salts and preparation methods, and their use as Fms-like tyrosine kinase 3 (FLT3), cell cycle checkpoint kinase 1 (CHK1) or dual-targeting FLT3 / CHK1 inhibitors in antitumor drugs. Background Technology

[0002] The FLT3 (FMS-like tyrosine kinase 3) gene is located on the long arm of human chromosome 13, region 1, band 2 (13q12), and is approximately 100 kb in length, comprising 24 exons, 16 of which are highly conserved with the c-KIT gene. It encodes a hematopoietic factor receptor tyrosine kinase—FLT3 kinase. FLT3 kinase belongs to the type III receptor tyrosine kinase family and is primarily expressed on CD34. + The surface of hematopoietic stem cells and immature hematopoietic progenitor cells plays a crucial role in regulating the survival, proliferation, and differentiation of hematopoietic progenitor cells. Aberrant activation of FLT3 is closely related to the development and progression of various tumors, particularly acute myeloid leukemia (AML). Studies have shown that FLT3 gene mutations account for approximately 30% of AML patients, making it the most common genetic mutation in AML. Depending on the mutation type, FLT3 mutations mainly include internal tandem repeat mutations located in the juxtamembrane domain (FLT3-ITD) and point mutations located in the kinase domain (FLT3-TKD). They account for 20% and 10% of AML patients, respectively. Both FLT3-ITD and FLT3-TKD can ligand-independently activate FLT3 and downstream signaling pathways, leading to the occurrence, development, and poor prognosis of AML. Based on the relationship between FLT3 mutations and AML, targeting FLT3 has become a research hotspot in AML.

[0003] Several FLT3 inhibitors are currently in clinical trials, and some have already been marketed. The use of FLT3 inhibitors has improved the survival and prognosis of AML patients to some extent. However, treatment responses are often short-lived, and the disease typically relapses within weeks, primarily due to mutations in the FLT3-kinase domain (acquired resistance) and activation of alternative pathways (adaptive resistance). Recent literature reports that type I FLT3 inhibitors can overcome most acquired resistance, while treatments that effectively overcome adaptive resistance are urgently needed.

[0004] Targeting the cell cycle has become a hot topic in anti-tumor research. DNA damage can cause cell cycle arrest in the G1, S, or G2 / M phases, with cell cycle checkpoint kinase 1 (CHK1) primarily responsible for regulating the S and G2 / M phases. Studies have shown that CHK1 overexpression is closely related to the development and progression of various tumors. In AML, high CHK1 expression is associated with shorter overall survival, progression-free survival, and recurrence-free survival. Furthermore, dual targeting of FLT3 and CHK1 can improve the treatment outcomes of AML patients with FLT3 mutations. Developing novel small-molecule inhibitors with excellent FLT3 / CHK1 kinase inhibitory activity is of great significance for the treatment of hematologic malignancies. Summary of the Invention

[0005] The purpose of this invention is to provide a class of N-type inhibitors with FLT3 and CHK1 inhibitory activities and antitumor effects. 2 -Replacing bicyclic-2-aminopyrimidine derivatives, these compounds exhibit excellent biological activity while enhancing molecular diversity and novelty; another object of this invention is to provide such N-containing compounds. 2 Preparation methods and applications of substituted bicyclic-2-aminopyrimidine derivatives.

[0006] The technical solution of the present invention is as follows:

[0007] Compounds of general formula (I) or their pharmaceutically acceptable salts:

[0008]

[0009] Or its optical isomer or its pharmaceutically acceptable salt.

[0010] in:

[0011] X represents NH and O;

[0012] A is a group represented by formula IA, IB, or IC:

[0013]

[0014] in Represents the substituent junction;

[0015] Y is selected from N and CR. X ;R X Selected from H, halogens, CH3, CF3;

[0016] m and n are independently selected from 0, 1, 2 or 3;

[0017] o can be selected from 0, 1, 2, 3, 4, or 5;

[0018] R1 is selected from -L-NRa R b -L-R8

[0019] L represents a chemical bond, C represents a chemical bond. 1-8 alkyl, C 3-10 The cycloalkyl group, wherein the group (alkyl, cycloalkyl) may be independently optionally surrounded by 0, 1, 2, or 3 R groups. 10 Replaced by multiple R 10 When replacing, multiple R 10 They can be independent and may be the same or different;

[0020] p and q are independently selected from 0, 1, 2, 3, 4, 5, 6, 7, and 8;

[0021] R a R b Independently selected from hydrogen, deuterium, hydroxyl, C 1-4 Alkyl, cyclopropyl, cyclobutyl, -C(=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 Alkyl groups);

[0022] W can be an acyl group, sulfonyl group, sulfoxide group, or ester group;

[0023] R2 is selected from C 1-5 alkyl, C 3-8 cycloalkyl, C 1-5 The alkoxy group (alkyl, cycloalkyl, alkoxy) can be independently surrounded by 0, 1, or 2 R groups. 11 Replaced by multiple R 11 When replacing, multiple R 11 They can be independent and may be the same or different;

[0024] R3, R4, R5, and R6 are independently selected from hydrogen, hydroxyl, halogen, and C. 1-3 The alkyl groups, or R3 and R4, R5 and R6, each independently form C10. 3-5 cycloalkyl or heterocycloalkyl;

[0025] R7 can be hydrogen, halogen, hydroxyl, methyl, or methoxy.

[0026] R8 is selected from hydrogen, hydroxyl, cyano, halogen, and C. 1-3 alkyl, C 1-3 Halogenated alkanes;

[0027] When R1 is selected from -L-R8, and o is 0, and L is cyclopropyl, R8 is not hydrogen;

[0028] R9 is selected from hydrogen, C 1-3 Alkyl, -C(=O)-(C 1-3Alkyl groups), -CHO, S(=O)2-(C 1-3 Alkyl groups);

[0029] R 10 It can be hydrogen, deuterium, halogen, hydroxyl, cyano, nitro, or C. 1-3 The alkoxy group; or the two R groups that substitute for each other on the same carbon atom. 10 Forming 3-6 membered rings;

[0030] R 11 Hydrogen, hydroxyl, halogen, nitro, cyano, C 1-4 alkyl, C 1-3 alkoxy, C 3-6 cycloalkyl groups;

[0031] A can be selected from any of the following structures:

[0032] The structures in IA, IB, and IC

[0033] Choose from any of the following structures:

[0034]

[0035] Preferably, it has the structure shown in general formula II:

[0036]

[0037] L represents a chemical bond, C represents a chemical bond. 1-8 alkyl, C 3-10 The cycloalkyl group, wherein the alkyl group or the cycloalkyl group may be independently and optionally surrounded by 0, 1, 2, or 3 R groups. 10 Replaced by multiple R 10 When replacing, multiple R 10 They can be independent and may be the same or different;

[0038] R a R b Independently selected from hydrogen, deuterium, hydroxyl, C 1-4 Alkyl, cyclopropyl, cyclobutyl, -C(=O)-(C 1-3 Alkyl groups), -C(=O)H, -S(=O)2-(C 1-3 Alkyl groups);

[0039] W can be an acyl group, a sulfonyl group, or an ester group;

[0040] R2 is selected from C 1-5 alkyl, C 3-8 cycloalkyl, C 1-5 The alkoxy group, wherein the alkyl, cycloalkyl, or alkoxy group can be independently surrounded by 0, 1, or 2 R groups. 11 Replaced by multiple R11 When replacing, multiple R 11 They can be independent of each other or they can be the same or different.

[0041] As a further preferred option, the R a R b Each element is independently selected from hydrogen, deuterium, -C(=0)-(C 1-3 Alkyl groups), -CHO, S(=O)2-(C 1-3 Alkyl group); W is -C(O)-; R2 is selected from C 1-5 The alkyl group can be further replaced by a hydroxyl group.

[0042] As a preferred embodiment, the compound has the following structure:

[0043] W is -C(O)-; R2 is selected from C 1-5 The alkyl group can be further replaced by a hydroxyl group.

[0044] In this invention, "alkyl" and the alkyl portion of other groups (e.g., alkoxy) can be straight-chain or branched, generally referring to C 1-5 Alkyl groups (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, etc.).

[0045] In this invention, "cycloalkyl" refers to a monocyclic or polycyclic hydrocarbon cyclic group, generally a 3-10 member cycloalkyl group, including, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclohexadienyl, cyclohepttrienyl, borneol, norpinyl, norcaryl, adamantyl, benzoyl, decahydronaphthyl, norbornyl, spiro[4.5]decyl, etc. The cycloalkyl group may be unsubstituted or substituted with one or more suitable substituents.

[0046] In this invention, "halogen" includes fluorine, chlorine, bromine or iodine, or fluoride, chloride, bromide or iodide.

[0047] In this invention, "heterocyclic alkyl" and "aromatic heterocyclic" refer to a ring system in which at least one carbon atom member is replaced by at least one heteroatom such as nitrogen, sulfur, or oxygen.

[0048] In this invention, "heterocyclic alkyl" refers to a non-aromatic monocyclic or polycyclic compound containing carbon and hydrogen atoms, as well as at least one heteroatom (preferably 1-4 heteroatoms from nitrogen, oxygen, sulfur, sulfone, or sulfoxide). Heterocyclic alkyl groups may have one or more carbon-carbon double bonds or carbon-heteroatom double bonds in the cyclic group, provided that the cyclic group does not become aromatic by its presence. The total number of carbon atoms and heteroatoms on the ring is generally 3-12.

[0049] In one embodiment of the present invention, X may specifically be NH or O.

[0050] In one embodiment of the present invention, Y may specifically be NH or CH.

[0051] In one embodiment of the present invention, R a R b Specifically, they can be selected independently from hydrogen, hydroxyl, methyl, methanesulfonyl, formyl, and acetyl.

[0052] In one embodiment of the present invention, W may be selected as an acyl group, a sulfonyl group, or an ester group.

[0053] In one embodiment of the present invention, R7 may specifically be hydrogen.

[0054] In one embodiment of the present invention, the structures in IA, IB, and IC... Specifically, it can be selected from any of the following structures:

[0055]

[0056] In one embodiment of the present invention, the compound of general formula I may be selected from the following specific compounds:

[0057]

[0058]

[0059]

[0060] The numbers below the above 52 compounds are their corresponding codes. For ease of description and brevity, the above codes will be used directly in the following content of this specification.

[0061] The present invention also provides the above-mentioned N 2 - A method for preparing substituted bicyclic-2-aminopyrimidine derivatives and their optical isomers or pharmaceutically acceptable salts thereof, achieved through the following steps:

[0062] The reaction route of the preparation method is shown in the following formula:

[0063]

[0064] Where Z = Br, Cl; the B ring is selected from:

[0065]

[0066] Any one of them; the definitions of R1, R2, R3, R4, R5, R6, R7, X, Y, m, n, and o are as described in claim 1; R1' is a Boc protected precursor of R1 or R1; the preparation method includes the following steps:

[0067] (3) Dissolve acidic compounds (such as acetic acid, glycolic acid, L-lactic acid, D-lactic acid, 2-methyl-2-hydroxypropionic acid, β-hydroxyisovaleric acid) in DMF, add HOBt, EDCI, compound 1 and DIPEA at 0℃ (e.g., an ice bath can be used), and carry out the reaction at 20℃~50℃ (e.g., the reaction can be carried out directly at room temperature). After the reaction is completed, compound 2 is obtained by post-treatment.

[0068] In step (3), after the reaction is detected by TLC, the reaction solution is extracted with ethyl acetate and washed with saturated brine, then the solvent is removed by vacuum distillation and silica gel column chromatography is performed to finally obtain compound 2.

[0069] (4) Dissolve compounds 2 and 3 in a reaction solvent, and under the protection of an inert gas (such as nitrogen), add Pd2(dba)3, Xant-phos and Cs2CO3, and react at 100℃~120℃ (for example, at 110℃). After the reaction is completed, process to obtain compound I or the Boc protected precursor of compound I, wherein the Boc protected precursor is further de-Boc grouped with trifluoroacetic acid to obtain compound I.

[0070] In step (4), after the reaction is completed, the solvent is removed by vacuum distillation and purified by silica gel column chromatography to obtain compound I or the Boc protected precursor of compound I.

[0071] Specifically, the following methods can be used:

[0072] Method 1: N 2 The synthetic route for substituted bicyclic-2-aminopyrimidine derivatives is as follows:

[0073]

[0074] The above N 2 The preparation method of substituted bicyclic-2-aminopyrimidine derivatives includes the following steps:

[0075] 1) Raw material 1 reacts with the corresponding acid at 20℃~50℃ under HOBt, EDCI and DIPEA conditions overnight (for example, it can be reacted directly at room temperature) to obtain intermediate 1, with DMF as the reaction solvent;

[0076] 2) Raw material 2 was reacted at low temperature for 1 hour under ammonia water conditions to obtain intermediate 2. The reaction solvent was acetonitrile, and the reaction temperature was an ice bath.

[0077] 3) Intermediate 2 reacts with the corresponding alcohol under alkaline conditions at room temperature overnight to give intermediate 3. The base used is potassium bis(trimethylsilyl)amino, and the reaction solvent is either THF or DMF.

[0078] 4) Intermediate 3 and intermediate 1 undergo Buchwald-Hartwig coupling under N2 protection, base, palladium and corresponding ligand catalysis, followed by removal of the Boc protecting group to obtain compound I-1. The solvent used for Buchwald-Hartwig coupling is anhydrous dioxane, the base is cesium carbonate, the catalyst is Pd2(dba)3, and the ligand is Xantphos.

[0079] 5) Intermediate 2 reacts with the corresponding amine under alkaline conditions and is heated overnight to obtain intermediate 4. The reaction solvent is any one of methanol, dioxane or acetonitrile, the base used is any one of DIPEA or TEA, and the reaction temperature is 80℃.

[0080] 6) Intermediate 4 and intermediate 1 undergo Buchwald-Hartwig coupling under N2 protection, base, palladium and corresponding ligand catalysis to directly obtain compound I-2 or to obtain compound I-2 by removing the Boc protecting group. The solvent used for Buchwald-Hartwig coupling is anhydrous dioxane, the base is cesium carbonate, the catalyst is Pd2(dba)3, and the ligand is Xantphos.

[0081] 7) Compound I-2 containing a primary or secondary amine (defined as compound I-3) undergoes a condensation or substitution reaction with the corresponding acid or alkyl sulfonyl chloride under alkaline conditions to yield compound I-4. When condensing with an acid, the condensing agent is HOBt or EDCI, the base is DIPEA, the solvent is DMF, and the reaction is carried out overnight at room temperature. When substituting with an alkyl sulfonyl chloride, the base is TEA, the reaction solvent is THF, and the reaction is carried out overnight at 60°C.

[0082] Method 2:

[0083] The compound of the present invention was dissolved in isopropanol, and a solution containing 1 to 1.5 times (e.g., 1.2 times equivalent) of an organic or inorganic acid was slowly added dropwise at room temperature. After the addition was complete, the reaction solution was stirred overnight at 40°C. The reaction solution was cooled to room temperature, filtered, and the solid was washed with diethyl ether and dried to obtain the corresponding salt.

[0084] In the above preparation process, Z is a chlorine or bromine atom, and R1, R2, L, R b As defined above, R' is R1 or the group before the removal of the Boc protecting group from R1, and the B ring is selected from any of the following structures:

[0085]

[0086] Those skilled in the art can prepare the compounds of the present invention using various starting compounds conventionally obtained in the art as raw materials, according to actual preparation needs.

[0087] This invention also provides a method for preparing the above-mentioned N 2 -A key intermediate in the process of substituted bicyclic-2-aminopyrimidine derivatives and their optical isomers or pharmaceutically acceptable salts thereof, with the structure shown below:

[0088]

[0089] A second object of the present invention is to provide a composition containing the above-mentioned 5-trifluoromethyl-pyrimidine-2,4-diamine compound.

[0090] In one embodiment of the invention, the composition comprises a 5-trifluoromethylpyrimidine-2,4-diamine compound of general formula I or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient, carrier, and diluent.

[0091] In one embodiment of the present invention, pharmaceutically acceptable salts are organic and inorganic salts. Organic salts include acetates, formates, propionates, pyruvates, glycolates, oxalates, oxalates, malates, succinates, glutarates, mandelates, citrates, trifluoroacetates, fumarates, oxalates, malates, L-malates, D-malates, lactates, camphor sulfonates, p-toluenesulfonates, methanesulfonates, ethanesulfonates, benzenesulfonates, salicylates, benzoates, tartrates, L-tartrates, D-tartrates, oxalates, succinates, maleates, ascorbic acid salts, and amino acid salts (such as aspartate). Inorganic salts include hydrochlorides, hydrobromides, sulfates, phosphates, nitrates, hydroiodates, and perchlorates.

[0092] In one embodiment of the invention, the compound or pharmaceutically acceptable salt may be prepared into a formulation by accepting an auxiliary ingredient. The formulation is a tablet, capsule, powder, granules, ointment, solution, suspension, injection, inhalation, gel, microsphere, or aerosol. The auxiliary ingredient may be, for example, cyclodextrin, arginine, or meglumine. The cyclodextrin is selected from α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, (C... 1-4 alkyl)-α-cyclodextrin, (C 1-4 alkyl)-β-cyclodextrin, (C 1-4 alkyl)-γ-cyclodextrin, (hydroxy-C) 1-4 alkyl)-α-cyclodextrin, (hydroxy-C) 1-4 alkyl)-β-cyclodextrin, (hydroxy-C) 1-4 alkyl)-γ-cyclodextrin, (carboxyl-C)1-4 alkyl)-α-cyclodextrin, (carboxyl-C) 1-4 alkyl)-β-cyclodextrin, (carboxyl-C) 1-4 Alkyl)-γ-cyclodextrin, α-cyclodextrin glycoethers, β-cyclodextrin glycoethers, γ-cyclodextrin glycoethers, α-cyclodextrin sulfonyl ether, β-cyclodextrin sulfonyl ether, and γ-cyclodextrin sulfonyl ether. The auxiliary components also include medically acceptable carriers, adjuvants, or mediators. Other pharmaceutically acceptable pharmaceutical compositions may include ion exchangers, alumina, aluminum stearate, and lecithin; buffering substances include phosphates, glycine, arginine, sorbic acid, etc.

[0093] In one embodiment of the invention, the invention provides pharmaceutical compositions comprising a compound of formula (I) and a pharmaceutically acceptable carrier, which can be formulated for intravenous administration, subcutaneous administration, inhalation, oral administration, rectal administration, parenteral administration, intravitreal administration, intramuscular administration, intranasal administration, transdermal administration, topical administration, ocular administration, intraocular administration, oral administration, intratracheal administration, bronchial administration, or sublingual administration. In other embodiments, these pharmaceutical compositions are formulated as tablets, pills, capsules, liquids, inhalers, nasal spray solutions, suppositories, solutions, gels, emulsions, ointments, eye drops, or ear drops.

[0094] This invention provides the use of the above-mentioned compound, composition, or formulation as an Fms-like tyrosine kinase 3 (FLT3), cell cycle checkpoint kinase 1 (CHK1), or dual-targeting FLT3 / CHK1 inhibitor in antitumor drugs.

[0095] Furthermore, the present invention provides the use of the above-mentioned compound, composition, or formulation in an antitumor drug for the preparation of a drug for diseases that benefit from Fms-like tyrosine kinase 3 (FLT3), cell cycle checkpoint kinase 1 (CHK1), or dual-target FLT3 / CHK1 inhibition.

[0096] In one embodiment of the invention, the invention provides a method for treating a protein kinase-mediated disease or condition, comprising administering to an individual in need of such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition, or an agent thereof. The protein kinase includes, but is not limited to, FLT3 (including mutant forms FLT3-ITD, FLT3-D835Y / V / F / I / N, FLT3-F691L / I, FLT3-ITD-D835Y / V / F, FLT3-ITD-F691L / I, FLT3-N676D, FLT3-Y842H / R, FLT3-G697R, FLT3-D815H / N, FLT3-K633Q) and CHK1.

[0097] In one embodiment of the present invention, diseases associated with the aforementioned kinase include tumors and autoimmune diseases. The tumors mentioned include bladder cancer, breast cancer, colon cancer, kidney cancer, epidermal cancer, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, prostate cancer, or skin cancer; lymphatic spectrum hematopoietic tumors such as acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, pilocellular lymphoma, Burkett's lymphoma, bone marrow spectrum hematopoietic tumors, acute and chronic myeloid leukemia, acute and chronic myeloid leukemia, spinal dysplasia syndrome, promyeloid leukemia, thyroid follicular carcinoma, stromal tumors, fibrosarcoma, rhabdomyosarcoma, central or peripheral nervous system tumors, astrocytoma, neuroblastoma, glioma, schwannoma, melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, xanthokeratoma, thyroid follicular carcinoma, and Kaposi's sarcoma.

[0098] In one embodiment of the present invention, the immune disease is selected from arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Auder's thyroiditis, Graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuritis, acute infectious encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, psoriasis, graft-versus-host disease, transplantation, transfusion allergy, allergic reaction, type I hypersensitivity reaction, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis.

[0099] In one embodiment of the invention, the active compound of the invention or a pharmaceutically acceptable salt thereof may be used alone as the sole antitumor drug, or may be used in combination with other therapeutic agents. These other therapeutic agents are selected from IDH1 inhibitors, IDH2 inhibitors, Bcl-2 inhibitors, hypomethylating agents, and antimetabolites.

[0100] The compounds of this invention have FLT3 and CHK1 inhibitory activities, and exhibit proliferation-inhibiting activity against various leukemia cell lines. They are also effective against various AML mutations, such as internal tandem repeat mutations in the juxtamembrane domain and D835 point mutations in the activation loop of the kinase domain. This can overcome drug resistance caused by point mutations in clinical practice and can be applied in the preparation of antitumor drugs. Attached Figure Description

[0101] Figure 1 This describes the therapeutic effect of compound 19 on human acute myeloid leukemia MV-4-11NU / NU mouse xenografts. Detailed Implementation

[0102] The present invention will be further described in conjunction with the accompanying drawings and embodiments.

[0103] The preparation of compounds will be further illustrated in the following examples. These examples are for illustrative purposes only and do not limit the invention in any way.

[0104] Preparation Example 1: 1-(2-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthidin-6(5H)-yl)ethyl-1-one (Compound 1)

[0105] Step 1: Synthesis of 1-(2-chloro-7,8-dihydro-1,6-naphthidium-6(5H)-yl)ethyl-1-one (intermediate 1-1)

[0106]

[0107] 2-Chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride (300 mg, 1.80 mmol) was dissolved in DMF (8 mL). HOBt (246 mg, 1.80 mmol), EDCI (348 mg, 1.80 mmol), DIPEA (474 ​​mg, 3.65 mmol), and acetic acid (1.80 mmol) were added sequentially under ice bath conditions. The mixture was stirred overnight at room temperature. The reaction solution was extracted three times with ethyl acetate and water, washed three times with saturated brine, and the organic layer was dried over anhydrous sodium sulfate. The solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using PE:EA = 1:1 as the eluent to give a yellow oil (265 mg, 71%). 1 H NMR(500MHz,Chloroform-d)δ7.43(d,J=8.5Hz,1H),7.20(d,J=8.5Hz,1H),4.74(s,2 H), 3.78 (t, J=6.0Hz, 2H), 3.06 (t, J=6.0Hz, 2H), 2.21 (s, 3H); ESI-MS: m / z=211[M+H] + .

[0108] Step 2: Synthesis of 4-chloro-5-(trifluoromethyl)pyrimidin-2-amine (intermediates 1-2)

[0109]

[0110] 2,4-Dichloro-5-trifluoromethylpyrimidine (7.8 g, 35.94 mmol) was dissolved in 30 mL of acetonitrile. Ammonia (28 mL) was added dropwise over an ice bath for more than 45 min. After the addition was complete, the reaction was continued in an ice bath for another 30 min. The solvent was recovered by vacuum distillation to obtain the residue. The residue was purified by silica gel plate with PE:EA = 7:1 as the eluent to give 1-2 white solids (3116 mg, 44%). 1 H NMR (500MHz, DMSO-d6) δ8.56 (s, 1H), 7.96 (s, 2H); ESI-MS: m / z=198[M+H] + .

[0111] Step 3: Synthesis of tert-butyl ((1S,4S)-4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)carbamate (intermediates 1-3)

[0112]

[0113] At room temperature, intermediate 1-2 (2.14 g, 10.84 mmol) was dissolved in anhydrous methanol (40 mL), and triethylamine (1.65 g, 16.26 mmol) and N-Boc-cis-cyclohexanediamine (1.91 g, 13.55 mol) were added sequentially with stirring. The reaction mixture was refluxed overnight, and the solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using PE:EA = 2:1 as the eluent to give a white solid 1-3 (3666 mg, 90%). 1 H NMR(500MHz,Chloroform-d)δ8.03(d,J=1.0Hz,1H),5.46–5.13(br,2H),5.01(d,J=7.0Hz,1H),4.16–4.13(m, 1H),3.64(s,1H),1.81–1.75(m,4H),1.67–1.63(m,2H),1.53–1.50(m,2H),1.44(s,9H); ESI-MS: m / z=376[M+H] + .

[0114] Step 4: 1-(2-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthidin-6(5H)-yl)ethyl-1-one (Compound 1)

[0115]

[0116] Under nitrogen protection, a mixture of intermediate 1-1 (250 mg, 0.67 mmol), intermediate 1-3 (0.80 mmol), tris(dibenzylindeneacetone)palladium (97 mg, 0.17 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (61 mg, 0.067 mmol), and cesium carbonate (436 mg, 1.34 mmol) was added to anhydrous dioxane (8 mL). The mixture was reacted overnight at 110 °C. After filtration, the solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using PE:EA = 1:1 as the eluent to give 305 mg of a pale yellow solid. Dissolve it in DCM (2 ml), add 0.5 ml of trifluoroacetic acid dropwise under ice bath, react the mixture at room temperature for 5 h, and then recover the solvent by vacuum distillation to obtain a yellow crude product. Purify it by silica gel plate column chromatography with DCM:NH3 / EtOH (30:1-20:1) as the eluent to obtain a white solid (153 mg, 51%). 1 H NMR(500MHz, Methanol-d4)δ8.29(s,1H),8.15(t,J=8.5Hz,1H),7.40(d,J=8.5Hz,1H),4.64(s,1H),4.57(s,1H),4.20(m,1H),3.86(t,J=6.0Hz,1H), 3.76(t,J=6.0Hz,1H),2.92(t,J=6.0Hz,2H),2.86(m,1H),2.16(d,J=7.5H z,3H),1.91–1.79(m,2H),1.75(m,4H),1.39(m,2H); ESI-MS:m / z=450[M+1] + .

[0117] Preparation Example 2 1-(2-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthidin-6(5H)-yl)-2-hydroxy-2-methylpropane-1-one (Compound 2)

[0118] Step 1: 1-(2-chloro-7,8-dihydro-1,6-naphthid-6(5H)-yl)-2-hydroxy-2-methylpropane-1-one (2-1)

[0119]

[0120] According to the preparation method of intermediate 1-1, 2-methyl-2-hydroxypropionic acid was used instead of acetic acid to obtain a yellow oily substance (60%). 1H NMR(500MHz,Chloroform-d)δ7.41(d,J=8.5Hz,1H),7.18(d,J=8.5Hz,1H),4.80(s,2 H), 4.00 (t, J=6.0Hz, 2H), 3.04 (t, J=6.0Hz, 2H), 1.53 (s, 6H); ESI-MS: m / z=255[M+H] + .

[0121] Step 2: 1-(2-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthidin-6(5H)-yl)-2-hydroxy-2-methylpropane-1-one (Compound 2)

[0122]

[0123] A white solid (42%) was prepared using intermediates 1-3 and 2-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, Methanol-d4) δ8.19(d,J=1.0Hz,1H),8.14(d,J=8.5Hz,1H),7.47(d,J=8.5Hz,1H),4.62(s,2H),4. 26–4.23(m,2H),3.91(s,1H),3.04–2.82(m,3H),1.90–1.75(m,6H),1.46(q,J=4.5Hz,8H); ESI-MS:m / z=494[M+H] + .

[0124] Preparation Example 3: 1-(2-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthidin-6(5H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 3)

[0125] Step 1: 1-(2-chloro-7,8-dihydro-1,6-naphthid-6(5H)-yl)-3-hydroxy-3-methylbutane-1-one (3-1)

[0126]

[0127] According to the preparation method of intermediate 1-1, β-hydroxyisovaleric acid was used instead of acetic acid to obtain a yellow oily substance (72%). 1H NMR (500MHz, DMSO-d6) δ7.71(d,J=8.5Hz,1H),7.35(d,J=8.5Hz,1H),4.75(s,0.8H),4.66(s,1.2H),3.83(t,J=6.0Hz,1.2H),3.79(t,J=6 .0Hz,0.8H),2.92(t,J=6.0Hz,1.2H),2.82(t,J=6.0Hz,0.8H),2.56(s,1H),2.54(s,1H),1.19(s,3H),1.16(s,3H); ESI-MS:m / z=269[M+H] + .

[0128] Step 2: 1-(2-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthidin-6(5H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 3)

[0129]

[0130] A white solid (58%) was prepared using intermediates 1-3 and 3-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ10.42(s,1H),8.48(d,J=4.5Hz,1H),8.03(t,J=8.0Hz,1H),7.56(t,J =9.0Hz,1H),6.02(br,1H),4.86(s,1H),4.69(s,1H),4.60(s,1H),4.17–4.12(m,1H),3.84–3 .79(m,2H),3.32–3.26(m,1H),2.84(t,J=6.0Hz,1H),2.74(t,J=6.0Hz,1H),2.56(d,J=9.0Hz ,2H),1.92–1.88(m,2H),1.83–1.63(m,6H),1.20(s,3H),1.17(s,3H); ESI-MS:m / z=508[M+H] + .

[0131] Preparation Example 4: 1-(5-((4-((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)isoindololin-2-yl)ethane-1-one (Compound 4)

[0132] Step 1: 1-(5-bromoisoindoline-2-yl)ethane-1-one (intermediate 4-1)

[0133]

[0134] According to the preparation method of intermediate 1-1, replacing 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride with 5-bromoisodihydroindole yielded a yellow oil (84%). 1 H NMR(500MHz,Chloroform-d)δ7.45–7.38(m,2H),7.14(dd,J=17.5,8.0Hz,1H),4.87–4.64(m,4H),2.16(s,CH3,3H); ESI-MS:m / z=240[M+H] + .

[0135] Step 2: 1-(5-((4-(((1s,4s)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)isoindoline-2-yl)ethane-1-one (compound 4)

[0136]

[0137] Using intermediates 1-3 and 4-1 as raw materials, a white solid (60%) was prepared according to the method in step 4 of Example 1. 1 HNMR(500MHz,DMSO-d6)δ9.71(s,1H),8.20(s,1H),7.83(d,J=2.0Hz,0.5H),7.74(d,J=2.0Hz ,0.5H),7.63(dd,J=8.5,2.0Hz,0.5H),7.54(dd,J=8.5,2.0Hz,0.5H),7.23(t,J=8.5Hz,1H), 6.17(d,J=7.5Hz,1H),4.78(s,1H),4.75(s,1H),4.58(s,1H),4.54(s,1H),4.15–4.07(m,1H) ,3.00–2.93(m,1H),2.04(s,3H),1.92-1.82(m,2H),1.61-1.42(m,6H); ESI-MS:m / z=435[M+H] + .

[0138] Preparation Example 5: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)isoindoline-2-yl)-2-hydroxyethane-1-one (Compound 5)

[0139] Step 1: 1-(5-bromoisoindoline-2-yl)-2-hydroxyethane-1-one (intermediate 5-1)

[0140]

[0141] According to the preparation method of intermediate 1-1, 5-bromoisodihydroindole and hydroxyacetic acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (70%). 1 H NMR(500MHz,Chloroform-d)δ7.51–7.41(m,2H),7.21(d,J=8.5Hz,0.5H),7.15(d,J=8.5Hz,0 .5H),4.86(s,1H),4.82(s,1H),4.67(s,1H),4.63(s,1H),4.21(s,2H); ESI-MS: m / z=256[M+H] + .

[0142] Step 2: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)isoindoline-2-yl)-2-hydroxyethane-1-one (compound 5)

[0143]

[0144] Using intermediates 1-3 and 5-1 as raw materials, a white solid (58%) was prepared according to the method in step 4 of Example 1. 1 HNMR(500MHz,DMSO-d6)δ9.72(s,1H),8.20(s,1H),7.84(s,0.5H),7.74(s,0.5H),7.71–7.61(dd ,J=8.5,2.0Hz,0.5H),7.55(dd,J=8.5,2.0Hz,0.5H),7.26(d,J=8.5Hz,0.5H),7.22(d,J=8.5Hz, 0.5H),6.16(d,J=7.5Hz,1H),4.72(s,1H),4.68(s,1H),4.65(s,1H),4.61(s,1H),4.13(s,2H),4 .12–4.06(m,1H)2.99–2.96(m,1H),1.94–1.80(m,2H),1.62–1.48(m,6H); ESI-MS:m / z=451[M+H] + .

[0145] Preparation Example 6: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethylpyrimidin-2-yl)amino)isoindol-2-yl)-2-hydroxy-2-methylpropane-1-one (Compound 6)

[0146] Step 1: 1-(5-bromoisoindoquinolin-2-yl)-2-hydroxy-2-methylpropane-1-one (intermediate 6-1)

[0147]

[0148] According to the preparation method of intermediate 1-1, 5-bromoisodihydroindole and 2-methyl-2-hydroxypropionic acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (72%). 1 H NMR(500MHz,Chloroform-d)δ7.46–7.40(m,2H),7.18(d,J=8.5Hz,0.5H),7.12(d,J=8.5Hz,0.5H),5 .02(s,1H),4.98(s,1H),4.86(s,1H),4.82(s,1H),3.93(br,1H),1.54(s,6H); ESI-MS:m / z=284[M+H] + .

[0149] Step 2: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethylpyrimidin-2-yl)amino)isoindoline-2-yl)-2-hydroxy-2-methylpropane-1-one (Compound 6)

[0150]

[0151] Using intermediates 1-3 and 6-1 as raw materials, a white solid (59%) was prepared according to the method in step 4 of Example 1. 1 HNMR(500MHz,DMSO-d6)δ9.69(s,1H),8.20(t,J=9.0Hz,1H),7.88–7.80(m,1H),7.65–7.41(m,1H),7.24(t,J=7.5Hz,1H),6.17(br,1H),5.35(s,1H ),5.11(d,J=8.0Hz,2H),4.66(d,J=9.5Hz,2H),4.12(s,1H),2.98(s,1H), 2.00–1.80(m,2H),1.73–1.48(m,6H),1.36(s,6H); ESI-MS:m / z=479[M+H] + .

[0152] Preparation Example 7: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethylpyrimidin-2-yl)amino)isoindol-2-yl)-3-hydroxy-3-methylbutane-1-one (Compound 7)

[0153] Step 1: 1-(5-bromoisoindoline-2-yl)-3-hydroxy-3-methylbutane-1-one (Intermediate 7-1)

[0154]

[0155] According to the preparation method of intermediate 1-1, 5-bromoisodihydroindole and β-hydroxyisovaleric acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (82%). 1 H NMR(500MHz,Chloroform-d)δ7.50–7.39(m,2H),7.19(d,J=8.5Hz,0.5H),7.13(d,J=8.5Hz,0.5H),4.79 (d,J=6.5Hz,2H),4.78–4.74(m,2H),2.51(d,J=1.0Hz,2H),1.33(d,J=1.0Hz,6H); ESI-MS:m / z=298[M+H] + .

[0156] Step 2: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethylpyrimidin-2-yl)amino)isoindoline-2-yl)-3-hydroxy-3-methylbutane-1-one (Compound 7)

[0157]

[0158] Using intermediates 1-3 and 7-1 as raw materials, a white solid (44%) was prepared according to the method in step 4 of Example 1. 1 HNMR(500MHz,DMSO-d6)δ9.71(s,1H),8.19(s,1H),7.85(s,0.5H),7.73(s,0.5H), 7.64(d,J=8.5Hz,0.5H),7.54(d,J=8.5Hz,0.5H),7.23–7.18(m,1H),6.13(s,1H), 4.84(s,1H),4.79(s,1H),4.61(s,1H),4.58(s,1H),4.10(s,1H),2.97(s,1H),2.4 7(s,2H),1.90–1.82(m,2H),1.61–1.49(m,6H),1.22(s,6H); ESI-MS:m / z=493[M+H] + .

[0159] Preparation Example 8: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)indololin-1-yl)ethane-1-one (Compound 8)

[0160] Step 1: 1-(5-bromoindoline-1-yl)ethane-1-one (intermediate 1-14)

[0161]

[0162] According to the preparation method of intermediate 1-1, 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride was replaced with 5-bromodihydroindole to obtain a yellow oil (86%). 1 H NMR(500MHz,Chloroform-d)δ8.11–8.06(m,1H),7.32–7.28(m,2H),4.06(t,J=8.5Hz,2H),3.19(t,J=8.5Hz,2H),2.22(s,3H); ESI-MS:m / z=240[M+H] + .

[0163] Step 2: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)indololin-1-yl)ethane-1-one (Compound 8)

[0164]

[0165] Using intermediates 1-3 and 8-1 as raw materials, a white solid (39%) was prepared according to the method in step 4 of Example 1. 1 HNMR(500MHz,Chloroform-d)δ8.20–8.01(m,2H),7.72–7.32(m,3H),5.18(d,J=7.5Hz,1H),4.21–4.16(m,1H),4.05(t,J=8. 5Hz,2H),3.18(t,J=8.5Hz,2H),3.02–2.85(m,1H),2.21(s,3H),1.90–1.80(m,3H),1.79–1.68(m,5H); ESI-MS:m / z=435[M+H] + .

[0166] Preparation Example 9: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)indoline-1-yl)-3-hydroxy-3-methylbutane-1-one (Compound 9)

[0167] Step 1: 1-(5-bromoindoline-1-yl)-3-hydroxy-3-methylbutane-1-one (Intermediate 9-1)

[0168]

[0169] According to the preparation method of intermediate 1-1, 5-bromodihydroindole and β-hydroxyisovaleric acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (74%). 1 H NMR(500MHz,Chloroform-d)δ8.11(d,J=9.0Hz,1H),7.33–7.29(m,2H),4.05(t,J= 8.5Hz, 2H), 3.20 (t, J=8.5Hz, 2H), 2.55 (s, 2H), 1.35 (s, 6H); ESI-MS: m / z=298[M+H] + .

[0170] Step 2: 1-(5-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)indoline-1-yl)-3-hydroxy-3-methylbutane-1-one (Compound 9)

[0171]

[0172] Using intermediates 1-3 and 9-1 as raw materials, a white solid (42%) was prepared according to the method in step 4 of Example 1. 1 HNMR(500MHz,Chloroform-d)δ8.16(d,J=8.5Hz,1H),8.14–8.12(m,1H),7.56–7.27(m,3H),5.28–5.12(m,1H),4.20(s,1H),4.05(t,J= 8.5Hz,2H),3.20(t,J=8.5Hz,2H),2.98(s,1H),2.56(s,2H),1.87–1.85(m,3H),1.83–1.65(m,5H),1.35(s,6H); ESI-MS: m / z=493[M+H] + .

[0173] Preparation Example 10: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)indololin-1-yl)ethane-1-one (Compound 10)

[0174] Step 1: 1-(6-bromoindol-1-yl)ethane-1-one (intermediate 10-1)

[0175]

[0176] According to the preparation method of intermediate 1-1, 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride was replaced with 5-bromodihydroindole to obtain a yellow oil (95%). 1 H NMR(500MHz,Chloroform-d)δ8.43(d,J=2.0Hz,1H),7.16(dd,J=8.0,2.0Hz,1H),7.04(d,J=8 .0Hz,1H),4.17–3.97(m,2H),3.15(t,J=8.5Hz,2H),2.18(s,CH3,3H); ESI-MS:m / z=240[M+H] + .

[0177] Step 2: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)indololin-1-yl)ethane-1-one (Compound 10)

[0178]

[0179] Using intermediates 1-3 and 10-1 as raw materials, a white solid (55%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.56(s,1H),8.46(s,1H),8.16(s,1H),7.32(d,J=8.0Hz,1H),7.09(d,J=8.0Hz,1H),6.04(d,J=8.0Hz,1H),4.23(s,1H),4.0 8(t,J=8.5Hz,2H),3.06(t,J=8.5Hz,2H),2.92(s,1H),2.14(s,3H),1.84– 1.72(m,2H),1.56–1.54(m,4H),1.47–1.41(m,2H); ESI-MS:m / z=435[M+H] + .

[0180] Preparation Example 11: 1-(7-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)ethane-1-one (Compound 11)

[0181] Step 1: 1-(7-bromo-3,4-dihydroisoquinoline-2(1H)-yl)ethane-1-one (intermediate 11-1)

[0182]

[0183] According to the preparation method of intermediate 1-1, 2-chloro-5,6,7,8-tetrahydro-1,6-naphthidine hydrochloride was replaced with 7-bromo-1,2,3,4-tetrahydroisoquinoline to obtain a yellow oil (56%). 1 H NMR(500MHz,Chloroform-d)δ7.33–7.27(m,2H),7.05–7.00(m,1H),4.69(s,1H),4.58(s,1H),3.80(t,J=6.0Hz,1H),3 .69–3.63(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.79(t,J=6.0Hz,1H),2.17(d,J=2.0Hz,3H); ESI-MS:m / z=254[M+H] + .

[0184] Step 2: 1-(7-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)ethane-1-one (Compound 11)

[0185]

[0186] Using intermediates 1-3 and 11-1 as raw materials, a white solid (66%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.07(d,J=7.5Hz,1H),7.68–7.52(m,2H),7.50–7.18 (m,2H),7.04–7.00(m,1H),5.15(d,J=7.5Hz,1H),4.65(s,1H),4.53(s,1H),4.16( s,1H),3.75(t,J=6.0Hz,1H),3.61(t,J=6.0Hz,1H),2.92(s,1H),2.80(t,J=6.0Hz ,1H),2.74(t,J=6.0Hz,1H),2.11(s,3H),1.87–1.61(m,8H); ESI-MS:m / z=449[M+H] + .

[0187] Preparation Example 12: 1-(7-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxy-2-methylpropane-1-one (Compound 12)

[0188] Step 1: 1-(7-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxy-2-methylpropane-1-one (Intermediate 12-1)

[0189]

[0190] According to the preparation method of intermediate 1-1, 7-bromo-1,2,3,4-tetrahydroisoquinoline and 2-methyl-2-hydroxypropionic acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (37%). 1 H NMR(500MHz,Chloroform-d)δ7.31(dd,J=8.5,2.0Hz,1H),7.28(d,J=2.0Hz,1H),7.02(d,J=8.5Hz, 1H), 4.78 (s, 2H), 3.88 (t, J=6.0Hz, 2H), 2.85 (t, J=6.0Hz, 2H), 1.53 (s, 6H); ESI-MS: m / z=298[M+H] + .

[0191] Step 2: 1-(7-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxy-2-methylpropane-1-one (Compound 12)

[0192]

[0193] Using intermediates 1-3 and 12-1 as raw materials, a white solid (59%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.12(s,1H),7.56–7.44(m,2H),7.33(dd,J=8.5,2. 5Hz,1H),7.07(d,J=8.5Hz,1H),5.23(d,J=7.0Hz,1H),4.81(s,2H),4.24–4.21(m ,1H),3.90(t,J=6.0Hz,2H),2.97–2.93(m,1H),2.87(t,J=6.0Hz,2H),1.89–1.85 (m,2H),1.78–1.73(m,4H),1.54(s,6H),1.45–1.35(m,2H); ESI-MS:m / z=493[M+H] + .

[0194] Preparation Example 13: 1-(7-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 13)

[0195] Step 1: 1-(7-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Intermediate 13-1)

[0196]

[0197] According to the preparation method of intermediate 1-1, 7-bromo-1,2,3,4-tetrahydroisoquinoline and β-hydroxyisovaleric acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (97%). 1 H NMR(500MHz,Chloroform-d)δ7.36–7.28(m,2H),7.05–7.02(m,1H),4.71(s,1H),4.58(s,1H),3.83(t,J=6.0Hz,1H),3.66 (t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.81(t,J=6.0Hz,1H),2.52(d,J=4.0Hz,2H),1.30(s,6H); ESI-MS: m / z=312[M+H] + .

[0198] Step 2: 1-(7-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 13)

[0199]

[0200] Using intermediates 1-3 and 13-1 as raw materials, a white solid (52%) was prepared according to the method in step 4 of Example 1. 1H NMR(500MHz,DMSO-d6)δ9.51(s,1H),8.07(s,1H),7.54(d,J=14.0Hz,1H),7.34(d,J=8.5 Hz,1H),6.94(t,J=6.0Hz,1H),6.01(s,1H),4.56(s,1H),4.49(s,1H),3.58(d,J=6.0Hz,2 H),2.85(s,1H),2.66(d,J=6.0Hz,1H),2.57(d,J=6.0Hz,1H),2.41–2.38(m,3H),1.76–1 .71(m,2H),1.51–1.46(m,4H),1.38(s,2H),1.06(d,J=5.5Hz,6H); ESI-MS:m / z=507[M+H] + .

[0201] Preparation Example 14: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)ethane-1-one (Compound 14)

[0202] Step 1: 1-(6-bromo-3,4-dihydroisoquinoline-2(1H)-yl)ethane-1-one (intermediate 14-1)

[0203]

[0204] According to the preparation method of intermediate 1-1, 2-chloro-5,6,7,8-tetrahydro-1,6-naphthidine hydrochloride was replaced with 6-bromo-1,2,3,4-tetrahydroisoquinoline to obtain a yellow oil (84%). 1 H NMR(500MHz,Chloroform-d)δ7.31–7.26(m,2H),6.99(d,J=8.0Hz,0.6H),6.98–6.94(d,J=8.0Hz,0.4H),4.64(s,1H),4.54(s,1H),3. 77(t,J=6.0Hz,1H),3.64(t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.80(t,J=6.0Hz,1H),2.16(d,J=2.5Hz,3H); ESI-MS: m / z=254[M+H] + .

[0205] Step 2: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)ethane-1-one (Compound 14)

[0206]

[0207] Using intermediates 1-3 and 14-1 as raw materials, a white solid (72%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.60(s,1H),8.19(s,1H),7.61(dd,J=10.0,2.0Hz,1H),7.53(dd,J=8.5,2.0Hz ,0.5H),7.48(dd,J=8.5,2.0Hz,0.5H),7.08(dd,J=8.5,4.0Hz,1H),6.13(d,J=7.5Hz,1H),4.57(s,1H), 4.52(s,1H),4.13(s,1H),3.64(t,J=6.0Hz,2H),2.97–2.95(m,1H),2.82(t,J=6.0Hz,1H),2.71(t,J=6 .0Hz,1H),2.07(s,3H),1.97–1.81(m,2H),1.64–1.52(m,4H),1.52–1.46(m,2H); ESI-MS: m / z=449[M+H] + .

[0208] Preparation Example 15: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxyethane-1-one (Compound 15)

[0209] Step 1: 1-(6-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxyethane-1-one (Intermediate 15-1)

[0210]

[0211] According to the preparation method of intermediate 1-1, 6-bromo-1,2,3,4-tetrahydroisoquinoline and hydroxyacetic acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (69%). 1H NMR (500MHz, DMSO-d6) δ7.43–7.35(m,2H),7.19(d,J=8.5Hz,0.5H),7.13(d,J=8.5Hz,0.5H),4.57(s,1H),4.51(s,1H),4.16(d, J=3.5Hz,2H),3.66(t,J=6.0Hz,1H),3.55(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.78(t,J=6.0Hz,1H); ESI-MS: m / z=270[M+H] + .

[0212] Step 2: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxyethane-1-one (Compound 15)

[0213]

[0214] Using intermediates 1-3 and 15-1 as raw materials, a white solid (74%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.82–9.50(m,1H),8.19(s,1H),7.62(d,J=8.0Hz,1H),7.51(d,J= 8.5Hz,1H),7.09(d,J=8.5Hz,1H),6.14(s,1H),4.56(s,1H),4.49(s,1H),4.17(s,2H),4. 17–4.11(m,1H)3.68(t,J=6.0Hz,1H),3.55(t,J=6.0Hz,1H),2.96(s,1H),2.81(t,J=6.0H z,1H),2.74(t,J=6.0Hz,1H),1.90–1.83(m,2H),1.64–1.42(m,6H); ESI-MS:m / z=465[M+H] + .

[0215] Preparation Example 16: (S)-1-(6-(((4-((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropane-1-one (Compound 16)

[0216] Step 1: (S)-1-(6-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropane-1-one (Intermediate 16-1)

[0217]

[0218] According to the preparation method of intermediate 1-1, 6-bromo-1,2,3,4-tetrahydroisoquinoline and L-lactic acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (60%). 1 H NMR(500MHz,Chloroform-d)δ7.38–7.27(m,2H),7.02(d,J=8.5Hz,0.6H),6.97(d,J=8.0Hz,0.4H),4.70(s,1H),4.56–4.44(m,2H),3.96–3.91(m ,0.5H),3.81–3.71(m,0.5H),3.69–3.56(m,1H),2.89(t,J=6.0Hz,2H), 1.37(d,J=6.5Hz,1.8H), 1.33(d,J=6.5Hz,1.2H); ESI-MS: m / z=284[M+H] + .

[0219] Step 2: (S)-1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropane-1-one (Compound 16)

[0220]

[0221] Using intermediates 1-3 and 16-1 as raw materials, a white solid (36%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.61(s,1H),8.18(s,1H),7.61(d,J=8.5Hz,1H),7.55–7.44(m,1H), 7.08(t,J=7.5Hz,1H),6.12(d,J=8.5Hz,1H),4.83–4.45(m,3H),4.20–4.07(m,1H),3.74(t, J=6.0Hz,1H),3.61(t,J=6.0Hz,1H),3.00–2.90(m,1H),2.82(t,J=6.0Hz,1H),2.74(t,J=6. 0Hz, 1H), 1.88–1.82 (m, 2H), 1.65–1.41 (m, 6H), 1.20 (d, J=6.5Hz, 3H); ESI-MS: m / z=479[M+H] + .

[0222] Preparation Example 17: (R)-1-(6-(((4-((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropane-1-one (Compound 17)

[0223] Step 1: (R)-1-(6-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropane-1-one (Intermediate 17-1)

[0224]

[0225] According to the preparation method of intermediate 1-1, 6-bromo-1,2,3,4-tetrahydroisoquinoline and D-lactic acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthylidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (92%). 1 H NMR(500MHz,Chloroform-d)δ7.36–7.29(m,2H),7.03(d,J=8.5Hz,1H),6.98(d,J=8.5Hz,0.5H),4.70(s,1H),4.61–4.46(m,2H), 3.98–3.90(m,0.5H),3.83–3.73(m,0.5H),3.66–3.55(m,1H),2.90(t,J=6.0Hz,2H),1.35(d,J=6.5Hz,3H); ESI-MS:m / z=284[M+H] + .

[0226] Step 2: (R)-1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropane-1-one (Compound 17)

[0227]

[0228] Using intermediates 1-3 and 17-1 as raw materials, a white solid (30%) was prepared according to the method in step 4 of Example 1. 1H NMR(500MHz,DMSO-d6)δ9.61(s,1H),8.18(s,1H),7.66–7.56(m,1H),7.50(dd,J=8.5, 2.0Hz,1H),7.08(t,J=7.5Hz,1H),6.12(d,J=8.5Hz,1H),4.78–4.47(m,3H),4.13(s,1H ),3.74(t,J=6.0Hz,2H),2.97–2.94(m,1H),2.82(t,J=6.0Hz,1H),2.74(t,J=6.0Hz,1 H),1.90–1.82(m,2H),1.64–1.43(m,6H),1.20(d,J=6.5Hz,3H); ESI-MS:m / z=479[M+H] + .

[0229] Preparation Example 18: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxy-2-methylpropane-1-one (Compound 18)

[0230] Step 1: 1-(6-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxy-2-methylpropane-1-one (intermediate 18-1)

[0231]

[0232] According to the preparation method of intermediate 1-1, 6-bromo-1,2,3,4-tetrahydroisoquinoline and 2-methyl-2-hydroxypropionic acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (75%). 1 H NMR(500MHz,Chloroform-d)δ7.35–7.30(m,2H),7.00(d,J=8.5Hz,1H),4.75(s,2H ),3.88(t,J=6.0Hz,2H),2.89(t,J=6.0Hz,2H),1.52(s,6H); ESI-MS:m / z=298[M+H] + .

[0233] Step 2: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxy-2-methylpropane-1-one (Compound 18)

[0234]

[0235] Using intermediates 1-3 and 18-1 as raw materials, a white solid (55%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.05(s,1H),7.57(s,1H),7.40(s,1H),7.30(d,J=8.5Hz,1H),6.98(d,J=8.5Hz,1H),5.15(d,J=6.5Hz,1H),4.72(s,2H ),4.14(s,1H),3.83(t,J=6.0Hz,2H),2.89(s,1H),2.82(d,J=6.0Hz,2H), 1.83–1.78(m,2H),1.70–1.65(m,6H),1.46(s,6H); ESI-MS:m / z=493[M+H] + .

[0236] Preparation Example 19: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 19)

[0237] Step 1: 1-(6-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Intermediate 19-1)

[0238]

[0239] According to the preparation method of intermediate 1-1, 6-bromo-1,2,3,4-tetrahydroisoquinoline and β-hydroxyisovaleric acid were used to replace 2-chloro-5,6,7,8-tetrahydro-1,6-naphthidine hydrochloride and acetic acid, respectively, to obtain a yellow oil (82%). 1 H NMR(500MHz,Chloroform-d)δ7.33–7.27(m,2H),7.00(d,J=8.5Hz,0.6H),6.95(d,J=8.5Hz,0.4H),5.16(s,1H),4.65(s,1H),4.53(s,1H),3.79(t,J =6.0Hz,1H),3.64(t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.81(t,J=6.0H z,1H),2.50(d,J=6.5Hz,2H),1.27(d,J=2.0Hz,6H); ESI-MS:m / z=312[M+H] + .

[0240] Step 2: 1-(6-((4-(((1S,4S)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 19)

[0241]

[0242] Using intermediates 1-3 and 19-1 as raw materials, a white solid (74%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.61(s,1H),8.18(s,1H),7.62(d,J=12.0Hz,1H),7.53–7.48(m,1H) ,7.07(dd,J=8.5,3.5Hz,1H),6.10(br,1H),4.65(s,1H),4.57(s,1H),4.13(s,1H),3.70(t, J=6.0Hz,2H),3.02–2.90(m,1H),2.81(t,J=6.0Hz,1H),2.72(t,J=6.0Hz,1H),2.53(d,J=5. 0Hz, 2H), 1.87–1.83 (m, 2H), 1.62–1.46 (m, 6H), 1.18 (d, J=9.0Hz, 6H); ESI-MS: m / z=507[M+H] + .

[0243] Preparation Example 20: N-((1S,4S)-4-((2-((2-(3-hydroxy-3-methylbutyryl)-1,2,3,4-tetrahydroisoquinoline-6-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)aminocyclohexyl)methanesulfonamide (Compound 20)

[0244]

[0245] Compound 19 (100 mg, 0.23 mmol) was dissolved in 3 mL of anhydrous tetrahydrofuran. Triethylamine (63 μL, 0.46 mmol) was added under ice bath conditions, followed by dropwise addition of 1 mL of anhydrous tetrahydrofuran solution containing methanesulfonyl chloride (59 mg, 0.46 mmol) over a period of more than 30 min. After the addition was complete, the reaction mixture was allowed to react at room temperature for 1 h, and then heated to 60 °C for 3 h. The solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using DCM:CH3OH = 40:1 as the eluent to give a white solid 20 (60 mg, 52%). 1H NMR(500MHz,Chloroform-d)δ8.14(s,1H),7.54(br,1H),7.49–7.35(m,2H),7.06(d,J=8.5Hz ,1H),5.20–5.13(br,1H),4.71(s,1H),4.58(s,1H),4.20–4.17(m,1H),3.84(t,J=6.0Hz,1H), 3.68(t,J=6.0Hz,1H),3.63–3.54(m,1H),3.01(s,3H),2.88(t,J=6.0Hz,1H),2.84(t,J=6.0H z,1H),2.54(d,J=6.0Hz,2H),2.00–1.57(m,8H),1.30(d,J=2.5Hz,6H); ESI-MS:m / z=585[M+H] + .

[0246] Preparation Example 21: N-((1S,4S)-4-((2-((2-(3-hydroxy-3-methylbutyryl)-1,2,3,4-tetrahydroisoquinoline-6-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)acetamide (Compound 21)

[0247]

[0248] Compound 19 (50 mg, 0.10 mmol) was dissolved in DMF (2 mL). HOBt (17 mg, 0.12 mmol), EDCI (24 mg, 0.12 mmol), acetic acid (7 mg, 0.12 mg), and DIPEA (30 mg, 0.23 mmol) were added sequentially under ice bath conditions. The mixture was reacted overnight at room temperature. After extraction with water and ethyl acetate 2-3 times, the organic layers were combined, washed 3 times with saturated brine, dried over anhydrous sodium sulfate, and the solvent was recovered under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography using DCM:CH3OH = 50:1 as the eluent to give a white solid 21 (35 mg, 65%). 1H NMR (500MHz, DMSO-d6) δ8.19–8.09(m,1H),7.91(d,J=8.5Hz,0.5H),7.68(d,J=8.5Hz,0.5H),7.54–7.43(m,2H),7.41– 7.35(m,1H),7.10(d,J=8.45Hz,0.5H),7.06(d,J=8.5Hz,0.5H),5.61(d,J=7.5Hz,1H),5.21(d,J=7.0Hz,1H),4.71(s, 1H),4.58(s,1H),4.23(s,1H),4.04–3.93(m,1H),3.84(t,J=6.0Hz,1H),3.68(t,J=6.0Hz,1H),2.89(t,J=6.0Hz,1H), 2.85(t,J=6.0Hz,1H),2.00(s,3H),1.94–1.75(m,6H),1.54–1.51(m,2H),1.31(d,J=3.0Hz,6H); ESI-MS:m / z=549[M+H] + .

[0249] Preparation Example 22: 1-(6-((4-(((1R,4R)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 22)

[0250] Step 1: tert-butyl((1R,4R)-4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)carbamate (intermediate 22-3)

[0251]

[0252] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with N-Boc-trans-cyclohexanediamine to obtain a white solid (92%). 1 H NMR(500MHz,Chloroform-d)δ8.04(d,J=1.0Hz,1H),5.08(s,2H),4.88(d,J=7.5Hz,1H),4.00–3.92( m,1H),3.51–3.45(m,1H),2.18–2.00(m,4H),1.40(s,9H),1.33–1.21(m,4H); ESI-MS:m / z=376[M+H] + .

[0253] Step 2: 1-(6-((4-(((1R,4R)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 22)

[0254]

[0255] Using intermediates 19-1 and 22-3 as raw materials, a white solid of 42% was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.57(s,1H),8.16(d,J=1.5Hz,1H),7.61–7.55(m,1.5H),7.47(dd,J=8.5,2.0Hz, 0.5H),7.09(dd,J=8.5,2.5Hz,1H),6.41(d,J=7.0Hz,1H),4.66(s,1H),4.58(s,1H),4.14–3.99(m,1H),3. 73(t,J=6.0Hz,1H),3.69(t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.74(t,J=6.0Hz,1H),2.54(d,J=9.0Hz ,3H),1.91–1.72(m,4H),1.60–1.44(m,2H),1.18(d,J=8.0,6H),1.15–1.08(m,2H); ESI-MS:m / z=507[M+H] + .

[0256] Preparation Example 23: (S)-3-hydroxy-3-methyl-1-(6-((4-(piperidin-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 23)

[0257] Step 1: (S)-3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 23-3)

[0258]

[0259] According to the preparation method of intermediates 1-3, replacing N-Boc cis-cyclohexanediamine with (S)-1-Boc-3-aminopiperidine yields a white solid (82%). 1H NMR(500MHz,Chloroform-d)δ8.06(s,1H),5.29–4.94(m,3H),4.16–4.14(m,1H),3.73–3.32(m,4H),1. 95(s,1H),1.87–1.81(m,1H),1.77–1.71(m,1H),1.69–1.63(m,1H),1.43(s,9H); ESI-MS: m / z=362[M+H] + .

[0260] Step 2: (S)-3-hydroxy-3-methyl-1-(6-((4-(piperidin-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (compound 23)

[0261]

[0262] Using intermediates 19-1 and 23-3 as raw materials, a white solid (62%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.15(s,1H),7.55(d,J=2.0Hz,0.5H),7.50(d,J=2.0Hz,0.5H),7.41–7.32(m,1H),7.26( s,1H),7.10(d,J=8.5Hz,0.5H),7.05(d,J=8.5Hz,0.5H),5.84(br,1H),4.71(s,1H),4.58(s,1H),4.22(d,J=8.5Hz,1H ),3.90–3.78(m,1H),3.67(t,J=6.0Hz,1H),3.12(t,J=6.0Hz,1H),2.91(t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.84– 2.82(m,2H),2.76–2.73(m,1H),2.53(d,J=8.5Hz,2H),1.87–1.64(m,4H),1.30(d,J=1.0Hz,6H); ESI-MS:m / z=493[M+H] + .

[0263] Preparation Example 24: (R)-3-hydroxy-3-methyl-1-(6-((4-(piperidin-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 24)

[0264] Step 1: (R)-3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 24-3)

[0265]

[0266] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with (R)-1-Boc-3-aminopiperidine to give a white solid (69%). 1 H NMR(500MHz,Chloroform-d)δ8.06(s,1H),5.29–4.94(m,3H),4.16–4.14(m,1H),3.73–3.32(m,4H),1. 95(s,1H),1.87–1.81(m,1H),1.77–1.71(m,1H),1.69–1.63(m,1H),1.43(s,9H); ESI-MS: m / z=362[M+H] + .

[0267] Step 2: (R)-3-hydroxy-3-methyl-1-(6-((4-(piperidin-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (compound 24)

[0268]

[0269] Using intermediates 19-1 and 24-3 as raw materials, a white solid (61%) was prepared according to the method in step 4 of Example 1. 1H NMR(500MHz,Chloroform-d)δ8.15(s,1H),7.55(d,J=2.0Hz,0.5H),7.50(d,J=2.0Hz,0.5H),7.37–7.33(m,1H),7.23(d, J=6.0Hz,1H),7.10(d,J=8.5Hz,0.5H),7.05(d,J=8.5Hz,0.5H),5.78(s,1H),4.71(s,1H),4.58(s,1H),4.32–4.16(m,1H ),3.84(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.13(d,J=11.5Hz,1H),2.91(t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.8 5–2.82(m,2H),2.76–2.73(m,1H),2.53(d,J=8.5Hz,2H),1.94–1.65(m,4H),1.31(d,J=1.0Hz,6H); ESI-MS:m / z=493[M+H] + .

[0270] Preparation Example 25: 3-Hydroxy-3-methyl-1-(6-((4-(piperidin-4-ylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 25)

[0271] Step 1: 4-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 25-3)

[0272]

[0273] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with N-Boc-4-aminopiperidine to give a white solid (80%). 1 H NMR(500MHz,Chloroform-d)δ8.05(d,J=1.0Hz,1H),5.12(s,2H),4.89(d,J=7.5Hz,1H),4.21–4.14(m,1H),4.10– 3.97(m,2H),2.92(s,2H),2.10(s,1H),2.06–1.96(m,2H),1.46(s,9H),1.40–1.37(m,1H); ESI-MS:m / z=362[M+H] + .

[0274] Step 2: 3-Hydroxy-3-methyl-1-(6-((4-(piperidin-4-ylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 25)

[0275]

[0276] Using intermediates 19-1 and 25-3 as raw materials, a white solid (58%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz, Methanol-d4)δ7.99(s,1H),7.39–7.34(m,2H),6.98(t,J=9.0Hz,1H),4.60(s,1H),4.55(s,1H),4.16–4.11(m,1H),3.68(t,J=6.0Hz,2H) ,2.98(d,J=12.5Hz,2H),2.76(t,J=6.0Hz,2H),2.58–2.52(m,4H),1.90–1. 84(m,2H),1.46–1.38(m,2H),1.20(d,J=1.0Hz,6H); ESI-MS:m / z=493[M+H] + .

[0277] Preparation Example 26: 3-Hydroxy-3-methyl-1-(6-((4-((piperidin-4-ylmethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 26)

[0278] Step 1: 4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)methyl)piperidine-1-carboxylic acid tert-butyl ester (intermediate 26-3)

[0279]

[0280] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with N-Boc-4-methylaminomethylpiperidine to obtain a white solid (98%). 1 H NMR(500MHz,Chloroform-d)δ8.03(s,1H),5.23(s,2H),5.14(d,J=6.5Hz,1H),4.11(s,2H),3.36(t,J=6.5Hz, 2H),2.68(s,2H),1.80–1.73(m,1H),1.70–1.64(m,2H),1.44(s,9H),1.18–1.10(m,2H); ESI-MS: m / z=376[M+H] + .

[0281] Step 2: 3-Hydroxy-3-methyl-1-(6-((4-((piperidin-4-ylmethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 26)

[0282]

[0283] Using intermediates 19-1 and 26-3 as raw materials, a white solid (57%) was prepared according to the method in step 4 of Example 1. 1 H NMR (500MHz, Methanol-d4) δ8.07(s,1H),7.52(dd,J=8.5,2.5Hz,1H),7.47(dd,J=6.5,2.5Hz,1H),7. 08(t,J=9.0Hz,1H),4.71(s,1H),4.66(s,1H),3.79(t,J=6.0Hz,2H),3.39(d,J=7.0Hz,2H),3.06–3.0 2(m,2H),2.90(t,J=6.0Hz,1H),2.84(t,J=6.0Hz,1H),2.65(d,J=6.5Hz,2H),2.55–2.50(m,2H),1.92 –1.87(m,1H),1.71(d,J=13.0Hz,2H),1.30(d,J=9.5Hz,6H),1.23–1.15(m,2H); ESI-MS:m / z=507[M+H] + .

[0284] Preparation Example 27: 1-(6-((4-((1S,2R)-2-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 27)

[0285] Step 1: tert-butyl((1R,2S)-2-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)carbamate (intermediate 27-3)

[0286]

[0287] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with (1R,2S)-2-aminocyclohexylcarbamate tert-butyl ester to obtain a white solid (87%). 1H NMR(500MHz,Chloroform-d)δ8.04(s,1H),5.52(s,1H),5.07(s,2H),4.33(s,1H),3.98 –3.93(m,1H),1.99–1.72(m,3H),1.65–1.47(m,5H),1.43(s,9H); ESI-MS: m / z=376[M+H] + .

[0288] Step 2: 1-(6-((4-(((1S,2R)-2-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 27)

[0289]

[0290] Using intermediates 19-1 and 27-3 as raw materials, a white solid of 62% was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.12(s,1H),7.53–7.35(m,3H),7.09(d,J=8.5Hz,0.5H),7.04(d,J=8.5Hz, 0.5H),6.19(br,1H),5.29(s,1H),4.71(s,1H),4.58(s,1H),4.13–4.08(m,1H),3.84(t,J=6.0Hz,1H),3.6 7(t,J=6.0Hz,1H),3.13(q,J=4.0Hz,1H),2.89(t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.53(d,J=9.0Hz,2 H),1.90–1.85(m,1H),1.69–1.65(m,3H),1.54–1.49(m,4H),1.30(d,J=1.0Hz,6H); ESI-MS:m / z=507[M+H] + .

[0291] Preparation Example 28: 1-(6-((4-((1R,2S)-2-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 28)

[0292] Step 1: tert-butyl((1S,2R)-2-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)carbamate (intermediate 28-3)

[0293]

[0294] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with (1S,2R)-2-aminocyclohexylcarbamate tert-butyl ester to obtain a white solid (75%). 1 H NMR(500MHz,Chloroform-d)δ8.04(s,1H),5.54(s,1H),5.07(s,2H),4.33(s,1H),3.97 –3.94(m,1H),2.00–1.72(m,3H),1.65–1.45(m,5H),1.43(s,9H); ESI-MS: m / z=376[M+H] + .

[0295] Step 2: 1-(6-((4-(((1R,2S)-2-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 28)

[0296]

[0297] Using intermediates 19-1 and 28-3 as raw materials, a white solid (69%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.12(s,1H),7.55–7.35(m,3H),7.10(d,J=8.5Hz,0.5H),7.04(d,J=8.5Hz, 0.5H),6.19(br,1H),5.29(s,1H),4.71(s,1H),4.58(s,1H),4.12–4.09(m,1H),3.84(t,J=6.0Hz,1H),3.6 7(t,J=6.0Hz,1H),3.13(q,J=4.0Hz,1H),2.89(t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.53(d,J=9.0Hz,2 H),1.96–1.81(m,1H),1.69–1.65(m,3H),1.59–1.44(m,4H),1.30(d,J=1.0Hz,6H); ESI-MS:m / z=507[M+H] + .

[0298] Preparation Example 29: (S)-3-hydroxy-3-methyl-1-(6-((4-((piperidin-3-ylmethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 29)

[0299] Step 1: Tert-butyl(R)-3-(((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)methyl)piperidine-1-carboxylic acid ester (intermediate 29-3)

[0300]

[0301] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with (R)-1-Boc-3-aminomethylpiperidine to obtain a white solid (95%). 1 H NMR(500MHz,Chloroform-d)δ8.04(s,1H),5.14(s,2H),3.92–3.63(m,2H),3.45–3.40(m,1H),3.37–3.31(m,1H),2. 97(s,1H),2.80–2.75(m,1H),1.85–1.80(m,2H),1.65(s,1H),1.44(s,9H),1.27–1.23(m,2H); ESI-MS: m / z=376[M+H] + .

[0302] Step 2: (S)-3-hydroxy-3-methyl-1-(6-((4-((piperidin-3-ylmethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (compound 29)

[0303]

[0304] Using intermediates 19-1 and 29-3 as raw materials, a white solid (59%) was prepared according to the method in step 4 of Example 1. 1H NMR(500MHz,Chloroform-d)δ8.13(t,J=1.0Hz,1H),7.52–7.39(m,3H),7.09(d,J=8.5Hz,0.5H),7.04(d,J=8.5Hz,0.5H),5.50(d, J=7.0Hz,1H),4.70(s,1H),4.58(s,1H),3.83(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.49–3.42(m,1H),3.41–3.36(m,1H),3.10– 3.04(m,1H),2.99–2.95(m,1H),2.89(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.62–2.57(m,1H),2.53(d,J=9.5Hz,2H),2.44–2.40 (m,1H),1.91–1.79(m,2H),1.72–1.68(m,1H),1.53–1.41(m,1H),1.30(d,J=1.0Hz,6H),1.23–1.15(m,1H); ESI-MS:m / z=507[M+H] + .

[0305] Preparation Example 30: (R)-3-hydroxy-3-methyl-1-(6-((4-((piperidin-3-ylmethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 30)

[0306] Step 1: (S)-3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)methyl)piperidine-1-carboxylic acid tert-butyl ester (30-3)

[0307]

[0308] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with (S)-1-Boc-3-aminomethylpiperidine to obtain a white solid (99%). 1 H NMR(500MHz,DMSO-d6)δ7.96(s,1H),6.93–6.64(m,3H),3.87–3.72(m,2H),3.28–3.17(m,2H),2.73–2.63(m,1H), 1.78(s,1H),1.72–1.65(m,1H),1.63–1.59(m,1H),1.46–1.24(m,11H),1.14–1.02(m,1H); ESI-MS:m / z=376[M+H] + .

[0309] Step 2: (R)-3-hydroxy-3-methyl-1-(6-((4-((piperidin-3-ylmethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (compound 30)

[0310]

[0311] Using intermediates 19-1 and 30-3 as raw materials, a white solid (55%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.18–8.10(m,1H),7.51–7.34(m,3H),7.10(d,J=8.5Hz,0.5H),7.04(d,J=8.5Hz,0.5H),5.50(s,1H), 4.71(s,1H),4.58(s,1H),3.84(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.51–3.43(m,1H),3.41–3.36(m,1H),3.10–3.06(m,1H),2.9 7(dt,J=12.0,4.0Hz,1H),2.90(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.62–2.57(m,1H),2.53(d,J=9.5Hz,2H),2.47–2.36(m,1H), 1.92–1.81(m,2H),1.71(dt,J=13.5,4.0Hz,1H),1.53–1.39(m,1H),1.30(d,J=1.0Hz,6H),1.25–1.14(m,1H); ESI-MS:m / z=507[M+H] + .

[0312] Preparation Example 31: 1-(6-((4-((2-aminoethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 31)

[0313] Step 1: tert-butyl (2-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)ethyl)carbamate (intermediate 31-3)

[0314]

[0315] According to the preparation method of intermediates 1-3, N-Boc cis-cyclohexanediamine was replaced with N-Boc ethylenediamine to obtain a white solid (99%).1 H NMR (500MHz, Chloroform-d) δ8.04(s,1H),5.78(s,1H),5.20(s,2H),3.54(q,J=6.0Hz,2H),3.37(q,J=6.0Hz,2H),1.43(s,9H); ESI-MS: m / z=322[M+H] + .

[0316] Step 2: 1-(6-((4-((2-aminoethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 31)

[0317]

[0318] Using intermediates 19-1 and 31-3 as raw materials, a white solid of 45% was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz, Methanol-d4)δ8.09(t,J=1.0Hz,1H),7.50(d,J=2.5Hz,1H),7.46–7. 43(m,1H),7.10(dd,J=8.4,5.5Hz,1H),4.71(s,1H),4.66(s,1H),3.79(t,J=6.0Hz ,2H),3.57(t,J=6.5Hz,2H),2.90(t,J=6.0Hz,1H),2.87(t,J=6.5Hz,2H),2.84(t, J=6.0Hz, 1H), 2.65 (d, J=6.0Hz, 2H), 1.30 (d, J=9.5Hz, 6H); ESI-MS: m / z=453[M+H] + .

[0319] Preparation Example 32: 1-(6-((4-((2-amino-2-methylpropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 32)

[0320] Step 1: tert-butyl(1-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)-2-methylpropane-2-yl)carbamate (intermediate 32-3)

[0321]

[0322] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with (2-amino-tert-butyl)carbamate tert-butyl ester to obtain a white solid (99%). 1 H NMR (500MHz, Chloroform-d) δ8.04(d,J=1.0Hz,1H),5.17(s,2H),5.08(s,1H),3.66(d,J=5.5Hz,2H),1.42(s,9H),1.30(s,6H); m / z=350[M+H] + .

[0323] Step 2: 1-(6-((4-((2-amino-2-methylpropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 32)

[0324]

[0325] Using intermediates 19-1 and 32-3 as raw materials, a white solid (45%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.15(s,1H),7.54(d,J=2.0Hz,0.5H),7.50(d,J=2.0Hz,0.5H),7.37(ddd,J=8.5, 6.5,2.0Hz,1H),7.26(s,1H),7.11(d,J=8.5Hz,0.5H),7.05(d,J=8.5Hz,0.5H),5.98(d,J=6.5Hz,1H),5.27(br ,1H),4.71(s,1H),4.58(s,1H),3.84(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.40(dd,J=5.0,1.5Hz,2H),2.90 (t,J=6.0Hz,1H),2.86(t,J=6.0Hz,1H),2.53(d,J=9.5Hz,2H),1.30(s,6H),1.18(s,6H); ESI-MS: m / z=481[M+H] + .

[0326] Preparation Example 33: 1-(6-((4-((3-aminopropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 33)

[0327] Step 1: tert-butyl(3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)propyl)carbamate (intermediate 33-3)

[0328]

[0329] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with N-Boc-propanediamine to obtain a white solid (94%). 1 H NMR(500MHz,Chloroform-d)δ8.04(s,1H),5.61(s,1H),5.24(s,2H),3.53(q,J=6.5H z,2H),3.18(q,J=6.5Hz,2H),1.79–1.65(m,2H),1.45(s,9H); ESI-MS:m / z=336[M+H] + .

[0330] Step 2: 1-(6-((4-((3-aminopropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 33)

[0331]

[0332] Using intermediates 19-1 and 33-3 as raw materials, a white solid (59%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.13(s,1H),7.53–7.39(m,2H),7.33(d,J=7.5Hz,1H),7 .10(d,J=8.5Hz,0.5H),7.04(d,J=8.5Hz,0.5H),6.72(s,1H),4.70(s,1H),4.57(s,1H ),3.83(t,J=6.0Hz,1H),3.68–3.62(m,3H),2.89(t,J=6.0Hz,3H),2.85(t,J=6.0Hz,1 H), 2.53 (d, J=10.0Hz, 2H), 1.77 (p, J=6.5Hz, 2H), 1.30 (s, 6H); ESI-MS: m / z=467[M+H] + .

[0333] Preparation Example 34: 1-(6-((4-((3-amino-2,2-dimethylpropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 34)

[0334] Step 1: tert-butyl(3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)-2,2-dimethylpropyl)carbamate (intermediate 1-41)

[0335]

[0336] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with N-Boc-amino-2,2-dimethyl-1,3-propanediamine to give a white solid (78%). 1 H NMR(500MHz,Chloroform-d)δ8.06(s,1H),5.12(s,1H),5.12(s,2H),3.32(d,J=7 .0Hz,2H),2.88(d,J=7.0Hz,2H),1.45(s,9H),0.90(s,6H); ESI-MS:m / z=364[M+H] + .

[0337] Step 2: 1-(6-((4-((3-amino-2,2-dimethylpropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 34)

[0338]

[0339] Using intermediates 19-1 and 34-3 as raw materials, a white solid (40%) was prepared according to the method in step 4 of Example 1. ¹H NMR (500MHz, Chloroform-d) δ 8.11 (s, 1H), 7.78 (s, 0.5H), 7.73 (s, 0.5H), 7.60 (d, J = 2.0Hz, 0.5H), 7.55 (d, J = 2.0Hz, 0.5H), 7.42–7.28 (m, 2H), 7.10 (d, J = 8.5Hz, 0.5H), 7.04 (d, J = 8.5Hz, 0.5H), 4.71 (s, 1H), 4.58 ( s,1H),3.84(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.47(dd,J=4.5,1.5Hz,2H),2.89(t,J=6.0Hz,1H),2. 85(t,J=6.0Hz,1H),2.68(s,2H),2.53(d,J=10.5Hz,2H),1.30(s,6H),0.97(s,6H); ESI-MS:m / z=495[M+H] + .

[0340] Preparation Example 35: 1-(6-((4-((3-amino-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 35)

[0341] Step 1: tert-butyl(4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)-2-methylbutane-2-yl)carbamate (intermediate 35-3)

[0342]

[0343] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with 3-N-Boc-3-methylbutane-1,3-diamine to give a white solid (95%). 1 H NMR(500MHz,Chloroform-d)δ8.05(s,1H),6.37(s,1H),5.41(s,2H),3.54-3. 42(m,2H),1.87-1.65(m,2H),1.48(s,9H),1.33(s,6H); ESI-MS:m / z=364[M+H] + .

[0344]

[0345] Using intermediates 19-1 and 35-3 as raw materials, a white solid (56%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.11(t,J=1.0Hz,1H),7.97(br,1H),7.53–7.46(m,1H),7.43(dd,J=8.5,2.0H z,1H),7.40–7.31(br,1H),7.09(d,J=8.5Hz,0.5H),7.04(d,J=8.5Hz,0.5H),5.28(br,1H),4.70(s,1H),4. 57(s,1H),3.83(t,J=6.0Hz,1H),3.66(t,J=6.0Hz,1H),3.65–3.61(m,2H),2.89(t,J=6.0Hz,1H),2.84(t,J =6.0Hz,1H),2.53(d,J=10.0Hz,2H),1.68(t,J=6.5Hz,2H),1.30(s,6H),1.20(s,6H); ESI-MS:m / z=495[M+H] + .

[0346] Preparation Example 36: 1-(6-((4-((4-aminobutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 36)

[0347] Step 1: Tert-butyl(4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)butyl)carbamate (intermediate 36-3)

[0348]

[0349] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with N-Boc-butanediamine to obtain a white solid (99%). 1 H NMR(500MHz,Chloroform-d)δ8.03(d,J=1.0Hz,1H),5.25(s,1H),5.13(s,1H),4.91(s,1H),3.48(q,J=6. 5Hz,2H),3.19(t,J=6.5Hz,2H),1.67–1.61(m,2H),1.56–1.50(m,2H),1.44(s,9H); ESI-MS:m / z=350[M+H] + .

[0350] Step 2: 1-(6-((4-((4-aminobutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 36)

[0351]

[0352] Using intermediates 19-1 and 36-3 as raw materials, a white solid of 42% was prepared according to the method in step 4 of Example 1. 1H NMR(500MHz,Chloroform-d)δ8.13(s,1H),7.49(d,J=2.5Hz,0.5H),7.46(d,J=2.5Hz,0.5H),7.43–7.40(m,1H),7 .33(d,J=7.5Hz,1H),7.10(d,J=8.5Hz,0.5H),7.05(d,J=8.5Hz,0.5H),5.81(br,1H),4.71(s,1H),4.58(s,1H),3 .84(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.59–3.50(m,2H),2.89(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.76 (t,J=6.5Hz,2H),2.53(d,J=9.0Hz,2H),1.76–1.68(m,2H),1.58–1.51(m,2H),1.30(s,6H); ESI-MS: m / z=481[M+H] + .

[0353] Preparation Example 37: 1-(6-((4-((5-aminopentyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 37)

[0354] Step 1: tert-butyl(5-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)pentyl)carbamate (intermediate 37-3)

[0355]

[0356] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with N-Boc-pentanediamine to obtain a white solid (99%). 1 H NMR(500MHz,Chloroform-d)δ8.01(d,J=1.0Hz,1H),5.30(br,2H),5.08(s,1H),3.50–3.36(m,2H),3.13(q, J=6.5Hz,2H),1.65–1.57(m,2H),1.55–1.49(m,2H),1.42(s,9H),1.40–1.33(m,2H); ESI-MS:m / z=364[M+H] + .

[0357] Step 2: 1-(6-((4-((5-aminopentyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 37)

[0358]

[0359] Using intermediates 19-1 and 37-3 as raw materials, a white solid (48%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.13(s,1H),7.54(s,1H),7.49(d,J=2.0Hz,0.5H),7.44(d,J=2.0Hz,0.5H),7.44–7.38(m ,1H),7.09(d,J=8.5Hz,0.5H),7.04(d,J=8.5Hz,0.5H),5.23(br,1H),4.70(s,1H),4.57(s,1H),3.83(t,J=6.0Hz,1H), 3.67(t,J=6.0Hz,1H),3.52(q,J=6.5Hz,2H),2.88(t,J=6.0Hz,1H),2.84(t,J=6.0Hz,1H),2.70(t,J=6.5Hz,2H),2.53( d,J=9.5Hz,2H),1.67(p,J=7.0Hz,2H),1.48(q,J=7.0Hz,2H),1.41(q,J=8.5Hz,2H),1.30(s,6H); ESI-MS: m / z=495[M+H] + .

[0360] Preparation Example 38: 1-(6-((4-((2-(1-aminocyclopropyl)ethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (Compound 38)

[0361] Step 1: Tert-butyl(1-(2-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)ethyl)cyclopropyl)carbamate (intermediate 38-3)

[0362]

[0363] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with N-[1-(2-aminoethyl)cyclopropyl]carbamate tert-butyl ester to obtain a white solid (98%). 1H NMR(500MHz,Chloroform-d)δ8.08(d,J=1.0Hz,1H),5.32(br,2H),3.62(td,J=6.5,5.0Hz,2H), 1.80(t,J=6.5Hz,2H),1.43(s,9H),0.51–0.46(m,2H),0.38–0.35(m,2H); ESI-MS:m / z=362[M+H] + .

[0364] Step 2: 1-(6-((4-((2-(1-aminocyclopropyl)ethyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one (compound 38)

[0365]

[0366] Using intermediates 19-1 and 38-3 as raw materials, a white solid (50%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.15(s,1H),7.50(s,1H),7.49(d,J=2.0Hz,0.5H),7.44(d,J=2.0Hz,0.5H),7.40–7 .33(m,1H),7.10(d,J=8.5Hz,0.5H),7.06(d,J=8.5Hz,0.5H),5.23(br,1H),4.70(s,1H),4.57(s,1H),3.83(t,J= 6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.62(td,J=6.5,5.0Hz,2H),2.80(t,J=6.0Hz,1H),2.75(t,J=6.0Hz,1H),2.54 (d,J=9.5Hz,2H),1.80(t,J=6.5Hz,2H),1.29(s,6H),0.51–0.46(m,2H),0.38–0.35(m,2H); ESI-MS: m / z=493[M+H] + .

[0367] Preparation Example 39: 3-Hydroxy-3-methyl-1-(6-((4-((3-(methylamino)propyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 39)

[0368] Step 1: tert-butyl(3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)propyl)(methyl)carbamate (intermediate 39-3)

[0369]

[0370] According to the preparation method of intermediates 1-3, N-(3-aminopropyl)-N-methylcarbamate tert-butyl ester was used to replace N-Boc-cis-cyclohexanediamine to obtain a white solid (98%). 1 H NMR(500MHz,DMSO-d6)δ7.96(s,1H),6.74(br,3H),3.73–3.68(m,2H),2.56–2.5 3(m,2H),2.24(s,3H),1.71(p,J=6.0Hz,2H),1.42(s,9H); ESI-MS:m / z=350[M+H] + .

[0371] Step 2: 3-Hydroxy-3-methyl-1-(6-((4-((3-(methylamino)propyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)butanone (Compound 39)

[0372]

[0373] Using intermediates 19-1 and 39-3 as raw materials, a white solid (69%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.54(s,1H),8.15(s,1H),7.76(s,1H),7.65(s,1H),7.51(d,J=8 .5Hz,1H),7.07(dd,J=8.5,4.5Hz,1H),4.90(s,1H),4.64(s,1H),4.56(s,1H),3.73–3.6 8(m,2H),3.50(t,J=6.0Hz,2H),2.81(t,J=6.0Hz,1H),2.73(t,J=6.0Hz,1H),2.56–2.53 (m,4H),2.24(s,3H),1.71(p,J=6.0Hz,2H),1.18(d,J=9.5Hz,6H); ESI-MS:m / z=481[M+H] + .

[0374] Preparation Example 40: 3-Hydroxy-1-(6-((4-((3-(isopropylamino)propyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbut-1-one (Compound 40)

[0375] Step 1: tert-butyl(3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)propyl)(isopropyl)carbamate (intermediate 40-3)

[0376]

[0377] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with N-(3-aminopropyl)-N-(propyl-2-yl)carbamate tert-butyl ester to obtain a white solid (90%). 1 H NMR(500MHz,Chloroform-d)δ8.28(s,1H),5.12(s,2H),5.08(s,1H),3.73–3.68(m,2H),3.55–3.51(m,1 H),2.56–2.53(m,2H),1.71(p,J=6.0Hz,2H),1.42(s,9H),1.23(d,J=7.5Hz,6H); ESI-MS:m / z=378[M+H] + .

[0378] Step 2: 3-Hydroxy-1-(6-((4-((3-(isopropylamino)propyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbutane-1-one (Compound 40)

[0379]

[0380] Using intermediates 19-1 and 40-3 as raw materials, a white solid (55%) was prepared according to the method in step 4 of Example 1. 1H NMR(500MHz,DMSO-d6)δ9.54(s,1H),8.15(s,1H),7.76(s,1H),7.65(s,1H),7.51(d,J=8.5Hz,1 H),7.07(dd,J=8.5,4.5Hz,1H),4.90(s,1H),4.64(s,1H),4.56(s,1H),3.73–3.68(m,2H),3.55 –3.51(m,1H),3.50(t,J=6.0Hz,2H),2.81(t,J=6.0Hz,1H),2.73(t,J=6.0Hz,1H),2.56–2.53(m ,4H),1.71(p,J=6.0Hz,2H),1.23(d,J=7.5Hz,6H), 1.18(d,J=9.5Hz,6H); ESI-MS:m / z=509[M+H] + .

[0381] Preparation Example 41: 3-Hydroxy-1-(6-((4-((3-(hydroxyamino)propyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbut-1-one (Compound 41)

[0382] Step 1: tert-butyl(3-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)propyl)(hydroxy)carbamate (intermediate 41-3)

[0383]

[0384] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with tert-butyl(3-aminopropyl)(hydroxy)carbamate to obtain a white solid (90%). 1 H NMR(500MHz,Chloroform-d)δ8.25(s,1H),5.12(s,2H),5.10(s,1H),3.75–3.66(m ,2H),2.56–2.53(m,2H),1.71(p,J=6.0Hz,2H),1.43(s,9H); ESI-MS:m / z=352[M+H] + .

[0385] Step 2: 3-Hydroxy-1-(6-((4-((3-(hydroxyamino)propyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbut-1-one (Compound 41)

[0386]

[0387] Using intermediates 19-1 and 41-3 as raw materials, a white solid (39%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.54(s,1H),8.15(s,1H),7.76(s,1H),7.65(s,1H),7.51(d, J=8.5Hz,1H),7.07(dd,J=8.5,4.5Hz,1H),4.90(s,1H),4.64(s,1H),4.56(s,1H),3.7 5–3.66(m,2H),3.50(t,J=6.0Hz,2H),2.81(t,J=6.0Hz,1H),2.73(t,J=6.0Hz,1H),2 .56–2.53(m,4H),1.71(p,J=6.0Hz,2H),1.18(d,J=9.5Hz,6H); ESI-MS:m / z=483[M+H] + .

[0388] Preparation Example 42: N-(3-((2-((2-(3-hydroxy-3-methylbutyryl)-1,2,3,4-tetrahydroisoquinoline-6-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)propyl)acetamide (Compound 42)

[0389]

[0390] According to Example 21, replacing compound 19 with compound 33 yielded a white solid (59%). 1 H NMR(500MHz, Methanol-d4)δ8.05(s,1H),7.51(d,J=2.5Hz,1H),7.44–7.41(m,1H),7.08(dd, J=8.5,3.5Hz,1H),4.67(s,2H),3.80(d,J=6.0Hz,1H),3.74(t,J=6.0Hz,1H),3.54(t,J=6.5Hz ,2H),3.23(t,J=6.5Hz,2H),2.90(t,J=6.0Hz,1H),2.84(t,J=6.0Hz,1H),2.59(d,J=8.0Hz,2 H),1.92(d,J=2.5Hz,3H),1.78(t,J=6.5Hz,2H),1.29(d,J=5.0Hz,6H); ESI-MS:m / z=509[M+H] + .

[0391] Preparation Example 43: N-(3-((2-((2-(3-hydroxy-3-methylbutyryl)-1,2,3,4-tetrahydroisoquinoline-6-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)aminopropyl)methanesulfonamide (Compound 43)

[0392]

[0393] According to Example 20, replacing compound 19 with compound 33 yielded a white solid (63%). 1 H NMR (500MHz, Methanol-d4) δ8.05(s,1H),7.47(d,J=2.5Hz,0.5H),7.45(d,J=2.5Hz,0.5H),7.41(dd,J=8.5,2.5Hz,0. 5H),7.39(dd,J=8.5,2.5Hz,0.5H),7.08(d,J=8.5Hz,0.5H),7.06(d,J=8.5Hz,0.5H),4.65(s,1H),4.59(s,1H),3.78(t ,J=6.0Hz,1H),3.69(t,J=6.0Hz,1H),3.62(t,J=6.5Hz,2H),3.10(t,J=6.5Hz,2H),2.89(d,J=6.0Hz,1H),2.87(d,J=2. 0Hz, 3H), 2.83 (t, J=6.0Hz, 1H), 2.54 (d, J=9.5Hz, 2H), 1.89–1.77 (m, 2H), 1.27 (d, J=2.5Hz, 6H); ESI-MS: m / z=545[M+H] + .

[0394] Preparation Example 44: N-(4-((2-((2-(3-hydroxy-3-methylbutyryl)-1,2,3,4-tetrahydroisoquinoline-6-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)-2-methylbutane-2-yl)acetamide (Compound 44)

[0395]

[0396] According to Example 21, replacing compound 19 with compound 35 yielded a white solid (70%). 1H NMR (500MHz, DMSO-d6) δ9.52(s,1H),8.15(s,1H),7.60(dd,J=7.0,3.0Hz,1H),7.51(dd,J=5.0,2.0Hz,1H),7.36(d, J=3.0Hz,1H),7.07(dd,J=8.5,5.0Hz,1H),6.92(s,1H),4.88(d,J=8.5Hz,1H),4.65(s,1H),4.56(s,1H),3.71(t,J=6 .0Hz,1H),3.67(t,J=6.0Hz,1H),3.45(q,J=7.0Hz,2H),2.81(t,J=6.0Hz,1H),2.72(t,J=6.0Hz,1H),2.53(d,J=7.0H z,2H),1.98–1.93(m,2H),1.75(d,J=2.0Hz,3H),1.25(d,J=2.0Hz,6H),1.17(d,J=1.2Hz,6H); ESI-MS: m / z=537[M+H] + .

[0397] Preparation Example 45: 3-Hydroxy-1-(6-((4-(((2-hydroxy-2-methylpropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbutane-1-one (Compound 45)

[0398] Step 1: 1-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)-2-methylpropane-2-ol (intermediate 45-3)

[0399]

[0400] According to the preparation method of intermediates 1-3, N-Boc-cis-cyclohexanediamine was replaced with 1-amino-2-methyl-2-propanol to obtain a white solid (90%). 1 H NMR (500MHz, DMSO-d6) δ8.01 (s, 1H), 6.82 (s, 2H), 5.92 (t, J = 5.5Hz, 1H), 4.82 (s, 1H), 3.34 (s, 2H), 1.10 (s, 6H); ESI-MS: m / z = 251 [M+H] + .

[0401] Step 2: 3-Hydroxy-1-(6-((4-(((2-hydroxy-2-methylpropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbutane-1-one compound 45)

[0402]

[0403] Under nitrogen protection, a mixture of intermediate 19-1 (0.67 mmol), intermediate 45-3 (0.80 mmol), tris(dibenzylindeneacetone)palladium (97 mg, 0.17 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (61 mg, 0.067 mmol), and cesium carbonate (436 mg, 1.34 mmol) was added to anhydrous dioxane (8 mL). The mixture was reacted overnight at 110 °C. After filtration, the solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using DCM:CH3OH = 30:1 as the eluent to give a white solid (66%). 1 HNMR(500MHz,Chloroform-d)δ8.16(s,1H),7.47(d,J=2.5Hz,0.5H),7.44(d,J=2.5Hz,0.5H),7.40(s,1H),7.33 (dt,J=8.5,2.5Hz,1H),7.10(d,J=8.5Hz,0.5H),7.05(d,J=8.5Hz,0.5H),5.66(d,J=6.0Hz,1H),5.28(d,J=13.5 Hz,1H),4.70(s,1H),4.58(s,1H),3.83(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),3.56(dd,J=5.5,2.0Hz,2H),2.8 9(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.53(d,J=9.5Hz,2H),1.30(s,6H),1.27(s,6H); ESI-MS: m / z=482[M+H] + .

[0404] Preparation Example 46: 3-Hydroxy-1-(6-((4-((3-hydroxy-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbutane-1-one (Compound 46)

[0405] Step 1: 4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)-2-methylbutane-2-ol (intermediate 46-3)

[0406]

[0407] According to the preparation method of intermediates 1-3, replacing N-Boc-ciscyclohexanediamine with 4-amino-2-methyl-but-2-ol yields a white solid (95%). 1H NMR(500MHz,Chloroform-d)δ8.02(d,J=1.0Hz,1H),6.22(s,1H),5.09(s,2H),3.6 0(td,J=6.5,5.0Hz,2H),1.78(t,J=6.5Hz,2H),1.30(s,6H); ESI-MS:m / z=265[M+H] + .

[0408] Step 2: 3-Hydroxy-1-(6-((4-((3-hydroxy-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbutane-1-one (Compound 46)

[0409]

[0410] According to the preparation method in step 2 of Example 45, intermediate 45-3 was replaced with intermediate 46-3 to obtain a white solid (57%). 1 H NMR(500MHz,Methanol-d4)δ8.03(s,1H),7.56–7.41(m,2H),7.17–7.02(m,1H), 4.67(s,2H),3.80(d,J=6.0Hz,1H),3.78–3.73(m,1H),3.64(t,J=7.0Hz,2H),2.9 1(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.61(dd,J=9.5,2.5Hz,2H),1.78(t,J =7.0Hz, 2H), 1.29 (d, J = 5.5Hz, 6H), 1.25 (d, J = 1.2Hz, 6H); ESI-MS: m / z = 496 [M+H] + .

[0411] Preparation Example 47: 1-(6-((4-((3-hydroxy-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)ethane-1-one (Compound 47)

[0412]

[0413] According to the preparation method in step 2 of Example 45, intermediates 45-3 and 19-1 were replaced by intermediates 46-3 and 14-1 respectively, to obtain a white solid (68%). 1H NMR (500MHz, Methanol-d4) δ8.06 (s, 1H), 7.56–7.51 (m, 1H), 7.49 (d, J = 2.0Hz, 1H), 7.11 (d,J=8.5Hz,0.5H),7.09(d,J=8.5Hz,0.5H),4.65(s,1H),4.63(s,1H),3.76(t,J=6.0Hz ,1H),3.73(t,J=6.0Hz,1H),3.64(t,J=7.5Hz,2H),2.92(t,J=6.0Hz,1H),2.84(t,J=6.0 Hz, 1H), 2.18 (d, J=3.5Hz, 3H), 1.79 (t, J=7.5Hz, 2H), 1.25 (s, 6H); ESI-MS: m / z=438[M+H] + .

[0414] Preparation Example 48: (S)-1-(6-((4-((2-fluoro-3-hydroxy-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbut-1-one (Compound 48)

[0415] Step 1: (S)-4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)-3-fluoro-2-methylbut-2-ol (intermediate 48-3)

[0416]

[0417] According to the preparation method of intermediates 1-3, N-Boc-ciscyclohexanediamine was replaced with (S)-4-amino-3-fluoro-2-methylbutane-2-ol to obtain a white solid (97%). 1 H NMR(500MHz,DMSO-d6)δ8.01(s,1H),6.82(s,2H),5.92(t,J=5.5Hz,1H),4.0 3–4.18(m,1H),2.93–2.71(m,2H),1.09–1.07(m,6H); ESI-MS:m / z=283[M+H] + .

[0418] Step 2: (S)-1-(6-((4-((2-fluoro-3-hydroxy-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutyl-1-one (Compound 48)

[0419]

[0420] According to the preparation method in step 2 of Example 45, intermediate 45-3 was replaced with intermediate 48-3 to obtain a white solid (77%). 1 H NMR(500MHz,DMSO-d6)δ9.52(s,1H),8.15(s,1H),7.60(dd,J=7.0,3.0Hz,1H),7.51(dd,J=5.0,2.0H z,1H),7.36(d,J=3.0Hz,1H),7.07(dd,J=8.5,5.0Hz,1H),6.92(s,1H),4.88(d,J=8.5Hz,1H),4.65( s,1H),4.18-4.03(m,1H),3.71(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),2.93–2.73(m,3H),2.72(t,J =6.0Hz,1H),2.53(d,J=7.0Hz,2H),1.25(d,J=1.2Hz,6H),1.09–1.07(m,6H); ESI-MS:m / z=514[M+H] + .

[0421] Preparation Example 49: (S)-1-(6-((4-((2-fluoro-3-hydroxy-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)ethyl-1-one (Compound 49)

[0422]

[0423] According to the preparation method in step 2 of Example 45, intermediates 45-3 and 19-1 were replaced by intermediates 48-3 and 14-1 respectively, to obtain a white solid (70%). 1 H NMR(500MHz,DMSO-d6)δ9.52(s,1H),8.15(s,1H),7.60(dd,J=7.0,3.0Hz,1H),7.51(dd,J=5 .0,2.0Hz,1H),7.36(d,J=3.0Hz,1H),7.07(dd,J=8.5,5.0Hz,1H),6.92(s,1H),4.88(d,J=8 .5Hz,1H),4.65(s,1H),4.18-4.03(m,1H),3.71(t,J=6.0Hz,1H),3.67(t,J=6.0Hz,1H),2.9 3–2.73(m,3H),2.72(t,J=6.0Hz,1H),2.07(s,3H),1.09–1.07(m,6H); ESI-MS:m / z=456[M+H] + .

[0424] Preparation Example 50: 3-Hydroxy-3-methyl-1-(6-((4-(((1-methylcyclopropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)but-1-one (Compound 50)

[0425] Step 1: N 4 -(1-Methylcyclopropyl)-5-(trifluoromethyl)pyrimidine-2,4-diamine (intermediate 50-3)

[0426]

[0427] According to the preparation method of intermediates 1-3, replacing N-Boc-cis-cyclohexanediamine with 1-methylcyclopropylamine yields a white solid (97%). 1 H NMR (500MHz, DMSO-d6) δ8.10(s,1H),6.88(s,2H),6.05(s,1H),1.33(s,3H),0.89–0.72(m,2H),0.47–0.43(m,2H); ESI-MS: m / z=233[M+H] + .

[0428] Step 2: 3-Hydroxy-3-methyl-1-(6-((4-(((1-methylcyclopropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)but-1-one (Compound 50)

[0429]

[0430] According to the preparation method in step 2 of Example 45, intermediate 45-3 was replaced with intermediate 50-3 to obtain a white solid (68%). 1H NMR (500MHz, DMSO-d6) δ9.52(s,1H),8.10(s,1H),7.60(dd,J=7.0,3.0Hz,1H),7.51(dd,J=5.0,2. 0Hz,1H),7.36(d,J=3.0Hz,1H),7.07(dd,J=8.5,5.0Hz,1H),4.88(s,1H),4.65(s,1H),3.71(t,J=6 .0Hz,1H),3.67(t,J=6.0Hz,1H),2.89(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.53(d,J=9.5Hz,2H ), 1.33(s,3H),1.25(d,J=1.2Hz,6H),0.89–0.72(m,2H),0.47–0.43(m,2H); ESI-MS:m / z=464[M+H] + .

[0431] Preparation Example 51: 3-Hydroxy-1-(6-((4-(((1-hydroxycyclopropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbut-1-one (Compound 51)

[0432] Step 1: 1-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclopropane-1-ol (intermediate 51-3)

[0433]

[0434] According to the preparation method of intermediates 1-3, replacing N-Boc-cis-cyclohexanediamine with 1-aminocyclopropanol yields a white solid (97%). 1 H NMR (500MHz, DMSO-d6) δ8.10(s,1H),6.88(s,2H),6.05(s,1H),6.02(s,1H),0.89–0.72(m,2H),0.47–0.43(m,2H); ESI-MS: m / z=235[M+H] + .

[0435] Step 2: 3-Hydroxy-1-(6-((4-((1-hydroxycyclopropyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-methylbut-1-one (Compound 51)

[0436]

[0437] According to the preparation method in step 2 of Example 45, intermediate 45-3 was replaced with intermediate 51-3 to obtain a white solid (50%). 1 H NMR (500MHz, DMSO-d6) δ9.52 (s, 1H), 8.10 (s, 1H), 7.60 (dd, J=7.0, 3.0Hz, 1H), 7.51 (d, J= 7.0Hz,1H),7.36(d,J=3.0Hz,1H),6.02(s,1H),4.88(s,1H),4.65(s,1H),3.71(t,J=6.0Hz ,1H),3.67(t,J=6.0Hz,1H),2.89(t,J=6.0Hz,1H),2.85(t,J=6.0Hz,1H),2.53(d,J=9.5H z,2H), 1.25(d,J=1.2Hz,6H).0.89–0.72(m,2H),0.47–0.43(m,2H); ESI-MS: m / z=466[M+H] + .

[0438] Preparation Example 52: 1-(6-((4-((1-aminocyclopropyl)methoxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbut-1-one (Compound 52)

[0439] Step 1: tert-butyl(1-(((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)oxy)methyl)cyclopropyl)carbamate (intermediate 52-3)

[0440]

[0441] Dissolve bis(trimethylsilyl)amino potassium (3.5 mL, 3.35 mmol) in anhydrous THF (5 mL). Add dropwise 1 mL of tert-butyl(1-(hydroxymethyl)cyclopropyl)carbamate (343 mg, 1.83 mmol) in THF under ice bath conditions. After reacting for 1 h on ice, add 1-2 (300 mg, 1.52 mmol). The mixture is reacted overnight at room temperature. The solvent is recovered under reduced pressure to obtain the residue, which is purified by silica gel column chromatography using DCM:EA = 2:1 as the eluent to give a white solid (318 mg, 60%). 1 H NMR(500MHz,Chloroform-d)δ8.24(s,1H),5.35(br,2H),5.07(s,1H),4.41(s, 2H),1.43(s,9H),0.99–0.93(m,2H),0.80–0.74(m,2H); ESI-MS:m / z=349[M+H] + .

[0442] Step 2: 1-(6-((4-((1-aminocyclopropyl)methoxy)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbut-1-one (compound 52)

[0443]

[0444] Using intermediates 19-1 and 52-3 as raw materials, a white solid (40%) was prepared according to the method in step 4 of Example 1. 1 H NMR(500MHz,Chloroform-d)δ8.38(d,J=2.5Hz,1H),7.52–7.41(m,2H),7.39(dd,J=8.5,2.5Hz,0.5H),7.35(dd ,J=8.5,2.5Hz,0.5H),7.13(d,J=8.5Hz,0.5H),7.08(d,J=8.5Hz,0.5H),5.28(br,1H),4.72(s,1H),4.59(s,1H ),4.33(d,J=1.5Hz,2H),3.84(t,J=6.0Hz,1H),3.68(t,J=6.0Hz,1H),2.90(t,J=6.0Hz,1H),2.85(t,J=6.0Hz, 1H),2.53(d,J=7.5Hz,2H),1.30(d,J=1.5Hz,6H),0.80–0.72(m,2H),0.69–0.61(m,2H); ESI-MS:m / z=480[M+H] + .

[0445] Preparation Example 53: Salt-forming compound of 1-(6-((4-((3-amino-3-methylbutyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroisoquinoline-2(1H)-yl)-3-hydroxy-3-methylbutane-1-one

[0446]

[0447] acid = citric acid, hemifumaric acid, malic acid, L-malic acid, D-malic acid, methanesulfonic acid, L-tartaric acid, D-tartaric acid, succinic acid

[0448] Succinic acid, maleic acid, formic acid, acetic acid, hydrochloric acid, phosphoric acid

[0449] Compound 19 was dissolved in isopropanol, and a solution containing 1.2 equivalents of organic or inorganic acid was slowly added dropwise at room temperature. After the addition was complete, the reaction solution was stirred overnight at 40°C. The reaction solution was cooled to room temperature, filtered, and the solid was washed with diethyl ether and dried to obtain the corresponding salts (53, citrate of compound 19; 54, hemifumarate of compound 19; 55, malate of compound 19; 56, L-malate of compound 19; 57, D-malate of compound 19; 58, methanesulfonate of compound 19; 59, L-tartrate of compound 19; 60, D-tartrate of compound 19; 61, succinate of compound 19; 62, maleate of compound 19; 63, formate of compound 19; 64, acetate of compound 19; 65, hydrochloride of compound 19; 66, phosphate of compound 19).

[0450] Bioactivity testing section

[0451] Example 54. Kinase Activity Assay

[0452] The inhibitory activity of the compounds provided in this invention against FLT3, FLT3-D835Y and CHK1 kinases was tested.

[0453] Instrument: Envision microplate reader TM (PerkinElmer, USA).

[0454] Materials: Human recombinant FLT3, purchased from Carna Biosciences, FLT3 protein fused with GST (aa564-993), and an FLT3 activity assay kit. KinEASE TM TK was purchased from Cisbio; human recombinant FLT3-D835Y was purchased from Eurofins; the FLT3 protein fragment fused with GST (containing the D835Y mutation) (aa564-end) was used; and an FLT3-D835Y activity assay kit was used. KinEASE TM TK was purchased from Cisbio; human recombinant CHK1 was purchased from Sino Biological, and was a CHK1 protein fragment fused with GST (aa1-476).

[0455] Sample preparation: Dissolve DMSO, store at low temperature, and control the concentration of DMSO in the final system within a range that does not affect the detection activity.

[0456] Experimental procedure: FLT3, FLT3-D835Y and the substrate (specifically biotin-labeled peptide TK Substrate) were diluted with HTRF kinase buffer (1.25X Kinase buffer, 6.25mM MgCl2, 1.25mM MnCl2, 1.25mM DTT), respectively. CHK1 and the substrate were diluted with HTRF kinase buffer (1X Kinase buffer, 5mM MgCl2, 1mM DTT). 4 μL of enzyme, 4 μL of substrate, and 2 μL of different concentrations of the test compound were added to a 384 reaction plate (ProxiPlate™-384Plus, PerkinElmer). The specific reaction systems were: FLT3: 2% DMSO, 0.5 ng / μL for eTM-3, 1.5 ng / μL for eTM-3, and 2.5 ng / μL for 25 ng / μL; FLT3-D835Y: 2% DMSO, 0.4 ng / μL for FLT3-D835Y, 1.4 ng / μL for 14 ng / μL; CHK1: 2% DMSO, 0.075 ng / FLT3-D, 1075 ng / μL, 50 7M ATP. After incubation at room temperature for 1 hour, antibodies were added for detection. A control group (using DMSO instead of the test compound as solvent) and a blank control group were also included. Each sample and each concentration was tested in triplicate. Initial screening was performed under single concentration conditions, for example, 10-selection, to test the activity of the samples. For samples that exhibit activity under certain conditions, such as an inhibition rate (%inhibition) greater than 50%, the activity dose-dependent relationship, i.e., IC50, is tested. 50 The value was obtained by nonlinearly fitting the sample concentration to the sample activity. The calculation software used was Graphpad Prism 8, and the fitting model used was sigmoidal dose-response (variable slope). For most inhibitor screening models, the bottom and top of the fitting curve were set to 0 and 100, respectively.

[0457] Table 1. Inhibitory activity of the compounds of the present invention against FLT3, FLT3-D835Y, and CHK1 kinases.

[0458]

[0459]

[0460]

[0461] IC 50 : Half-maximal inhibitory concentration

[0462] A: IC 50 <20nM; B:20nM<IC 50 <100nM; C:100nM<IC50 <1000nM; D:

[0463] IC 50 >1000nM

[0464] The results in Table 1 show that most of the compounds exhibit good FLT3 and CHK1 kinase inhibitory activity, indicating that these compounds have the potential to treat FLT3, CHK1, and FLT3 / CHK1-related diseases.

[0465] Example 55. Inhibitory activity test for cell proliferation

[0466] Cell lines: MV-4-11 (human acute myeloid monocytic leukemia, expressing FLT3-ITD homozygous mutation), Molm-13 (human acute myeloid monocytic leukemia, FLT3-ITD heterozygous mutation), BaF3-FLT3-ITD (human acute myeloid monocytic engineered cell line, expressing FLT3-ITD homozygous mutation).

[0467] Experimental procedure: The antiproliferative activity (IC50) of the test compound against MV-4-11 and other cell lines was determined by MTS assay. 50 Cells in logarithmic growth phase were digested with trypsin, counted, and seeded at a density of 1×10⁴ cells / well in 96-well plates (100 cells per well). The plates were incubated overnight at 37°C with 5% CO₂. Six concentration gradients were set up for each test compound, with three replicates for each concentration. After incubation for 72 h, 20 μL MTS was added. After incubation at 37°C for 2 h, the absorbance at 490 nm (L1) was measured using a SpectraMAX 340 microplate reader, with a reference wavelength of 690 nm (L2). The (L1-L2) values ​​were plotted against different inhibitor concentrations, and the IC50 of the compounds was calculated using GraphPad Prism 5 software. 50 (Inhibition rate = (OD value of control group - OD value of treatment group) / OD value of control group * 100%)

[0468] Table 2. Inhibitory activity of the compounds of the present invention against acute myeloid leukemia cells MV-4-11.

[0469] Compound numbering MV4-11 Compound numbering MV4-11 1 A 2 A 3 B 4 A 5 A 6 A 7 A 8 A 9 A 10 B 11 A 12 A 13 A 14 A 15 A 16 A 17 A 18 A 19 A 20 A 21 A 24 A 32 B 33 A 34 B 35 A 36 A 37 B 38 A 39 A 41 A 42 A 43 A 46 A 47 A 48 A 49 A 51 A 52 A 53 A 54 A 55 A 56 A 57 A 58 A 59 A 60 A 61 A 62 A 63 A 64 A 65 A 66 A CCT245737 C

[0470] IC 50 : Half-maximal inhibitory concentration

[0471] A: IC 50 <10nM; B:10nM<IC 50 <100nM; C:100nM<IC 50 <1000nM; D:IC 50 >1000nM

[0472] CCT245737:

[0473] Table 3. Inhibitory activity of the compounds of the present invention against Molm-13 acute myeloid leukemia cells.

[0474] Compound numbering Molm-13 Compound numbering Molm-13 1 B 2 B 3 B 4 A 5 A 6 A 7 A 8 A 9 A 10 B 11 A 12 A 13 A 14 A 15 A 16 A 17 A 18 A 19 A 20 B 21 B 24 A 33 B 35 B 36 B 37 B 38 A 53 A 54 A 55 A 56 A 57 A 58 A 59 A 60 A 61 A 62 A 63 A 64 A 65 A 66 A CCT245737 C

[0475] IC 50 : Half-maximal inhibitory concentration

[0476] A: IC 50 <10nM; B:10nM<IC 50 <100nM; C:100nM<IC 50 <1000nM; D:IC 50 >1000nM

[0477] Table 4. Inhibitory activity of the compounds of the present invention against BaF3-FLT3-ITD in acute myeloid leukemia cells.

[0478] Compound numbering BaF3-FLT3-ITD Compound numbering BaF3-FLT3-ITD 4 A 5 A 6 B 7 A 8 A 9 A 10 B 11 A 12 B 13 A 14 A 15 A 16 A 17 A 18 A 19 A 20 A 21 B 24 A 33 B 35 A 36 A 37 B 38 A 53 C 54 A 55 A 56 A 57 A 58 A 59 A 60 A 61 A 62 A 63 A 64 A 65 A 66 A CCT245737 C

[0479] IC 50 : Half-maximal inhibitory concentration

[0480] A: IC 50 <10nM; B:10nM<IC 50 <100nM; C:100nM<IC 50 <1000nM; D:IC 50 >1000nM

[0481] Tables 2, 3, and 4 above show that the compounds of the present invention have good inhibitory activity against the proliferation of MV-4-11, Molm-13, and BaF3-FLT3-ITD cells.

[0482] Table 5. Inhibitory activity of the preferred compound 19 of the present invention against other cell lines.

[0483] Cell lines <![CDATA[IC 50 (nM)]]> Z-138 114.55 JeKo-1 260.03 MM.1S 1096.67 Rec-1 3424

[0484] The results showed that the preferred compound 19 of the present invention exhibited excellent anti-proliferative activity against Z-138 and JeKo-1 cell lines.

[0485] In vivo antitumor activity test

[0486] Example 56. MV-4-11 xenograft model

[0487] Experimental methods: NU / NU mice were subcutaneously injected with human MV-4-11 cells at a seeding density of 5 × 10⁶ cells / mL. 6 / only, until the tumor grows to 100-300mm 3 Subsequently, based on animal weight and tumor size, the animals were randomly divided into three groups: a 0.5% CMC-Na solvent control group, a cytarabine (AraC) control group, and a treatment group, with six animals in each group. Treatment began with daily administration of compound 19 at 1 mg / kg and 3 mg / kg. The control group received daily subcutaneous injections of cytarabine (20 mg / kg), while the negative control group received the same amount of solvent for 22 days. During the experiment, tumor volume was measured twice weekly, and mouse weight was also measured. The experimental results are as follows: Figure 1 As shown

[0488] from Figure 1 It can be seen that compound 19 provided by the invention has a significant inhibitory effect on tumor growth in human acute myeloid leukemia MV-4-11.

Claims

1. A compound, characterized in that, It has the structure shown in general formula (I): ； Or its pharmaceutically acceptable salt; in: X is selected from NH and O; A is a group represented by formula IB or formula IC; ； in" "Represents the substituent connection point; Y is selected from N and CR. X ;R X Selected from H; m and n are independently selected from 0 and 1; o can be selected from 0, 1, 2, 3, 4, or 5; R1 is selected from -L-NR a R b -L-R8 ; L represents a chemical bond, C represents a chemical bond. 1-8 alkylene, C 3-10 The cycloalkylene group, wherein the alkylene group or the cycloalkylene group may be independently and optionally surrounded by 0, 1, 2, or 3 R groups. 10 Replaced by multiple R 10 When replacing, multiple R 10 They can be independent and may be the same or different; p is selected from 1 and 2; q is selected from 1, 2, and 3; R a R b Independently selected from hydrogen, deuterium, hydroxyl, C 1-4 Alkyl, cyclopropyl, cyclobutyl, -C(=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 Alkyl groups); W is -C(O)-R2 selected from C 1-5 The alkyl group, which may be further replaced by a hydroxyl group, R3 and R4 are independently selected from hydrogen; R7 is hydrogen; R8 is selected from hydrogen, hydroxyl, cyano, halogen, and C. 1-3 alkyl, C 1-3 Halogenated alkanes; When R1 is selected from -L-R8, and o is 0, and L is cyclopropyl, R8 is not hydrogen; R9 is selected from hydrogen, C 1-3 Alkyl, -C(=O)-(C 1-3 Alkyl groups), -CHO, S(=O)2-(C 1-3 Alkyl groups); R 10 It can be hydrogen, deuterium, halogen, hydroxyl, cyano, nitro, or C. 1-3 The alkoxy group; or the two R groups that substitute for each other on the same carbon atom. 10 It forms a 3-6 quincunx ring.

2. The compound according to claim 1, characterized in that, The structure in IB and IC Choose from any of the following structures: 。 3. The compound according to claim 1, characterized in that, It has the structure shown in general formula (II): (II); The L and R a R b W and R2 are defined as having equal weight 1.

4. The compound according to claim 3, characterized in that, The L is C 4-8 The cycloalkylene group; the R a R b Each element is independently selected from hydrogen, deuterium, -C(=0)-(C 1-3 Alkyl groups), -CHO, S(=O)2-(C 1-3 Alkyl groups).

5. The compound according to claim 1, characterized in that, Selected from the following compounds: Or a pharmaceutically acceptable salt.

6. The compound of claim 1, characterized in that, Pharmaceutically acceptable salts include acid addition salts formed by compounds of general formula (I) with the following acids: formic acid, acetic acid, propionic acid, pyruvic acid, glycolic acid, oxalic acid, malonic acid, succinic acid, glutaric acid, mandelic acid, citric acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, L-malic acid, D-malic acid, lactic acid, camphor sulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, salicylic acid, benzoic acid, tartaric acid, L-tartaric acid, D-tartaric acid, oxalic acid, succinic acid, maleic acid, ascorbic acid, amino acids, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, hydroiodic acid, or perchloric acid.

7. A method for preparing the compound according to any one of claims 1-6, wherein the reaction route of the preparation method is shown in the following formula: ； in, Z = Br, Cl; ring B is selected from: ； Any one of them; the definitions of R1, R2, R3, R4, R7, X, Y, m, n, and o are as described in claim 1; R1' is a Boc protected precursor of R1 or R1; the preparation method includes the following steps: (1) Dissolve the acid compound in DMF, add HOBt, EDCI, compound 1 and DIPEA at 0℃, and react at 20℃~50℃. After the reaction is completed, compound 2 is obtained by post-treatment. (2) Dissolve compound 2 and compound 3 in the reaction solvent, add Pd2(dba)3, Xant-phos and Cs2CO3 under inert gas protection, and react at 100℃~120℃. After the reaction is completed, process to obtain compound I or the Boc protected precursor of compound I. The Boc protected precursor is further de-Boc grouped with trifluoroacetic acid to obtain compound I.

8. A key intermediate for preparing any one of the compounds of claims 1-6, characterized in that, The intermediate is selected from any of the following structures: 。 9. A pharmaceutical composition, characterized in that, It includes one or more compounds as described in any one of claims 1-6.

10. A pharmaceutical preparation, characterized in that, It includes one or more compounds as described in any one of claims 1-6.

11. The pharmaceutical preparation according to claim 10, characterized in that, The formulation is a tablet, capsule, powder, granule, ointment, solution, suspension, injection, inhaler, gel, microsphere or aerosol.

12. The use of a compound of any one of claims 1-6 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of clinical diseases associated with FLT3 and / or CHK1.

13. Use of a compound according to any one of claims 1-6 or a pharmaceutically acceptable salt thereof in the preparation of an antitumor drug or an immune disease drug.

14. The application according to claim 13, characterized in that, The tumors mentioned include bladder cancer, breast cancer, colon cancer, kidney cancer, epidermal cancer, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, prostate cancer, or skin cancer; lymphatic spectrum hematopoietic tumors, bone marrow spectrum hematopoietic tumors, acute and chronic myeloid leukemia, acute and chronic myeloid leukemia, spinal dysplasia syndrome, promyeloid leukemia, thyroid follicular carcinoma, stromal tumors, fibrosarcoma, rhabdomyosarcoma, central or peripheral nervous system tumors, astrocytoma, neuroblastoma, glioma, schwannoma, melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, xanthokeratoma, and Kaposi's sarcoma.

15. The application according to claim 13, characterized in that, The immune diseases mentioned are selected from arthritis, lupus, inflammatory bowel disease, Still's disease, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Auder's thyroiditis, Graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuritis, acute infectious encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, psoriasis, graft-versus-host disease, transfusion allergy, allergic reaction, type I hypersensitivity reaction, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis.

16. The application according to claim 14, characterized in that, The compound is used alone or in combination with one or more other therapeutic agents.

17. The application according to claim 16, characterized in that, The other therapeutic agents are selected from IDH1 inhibitors, IDH2 inhibitors, Bcl-2 inhibitors, hypomethylating agents, and antimetabolites.

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