Aminopyrimidine compound, pharmaceutical composition, application of aminopyrimidine compound and intermediate compound

Abstract

The invention relates to the technical field of compound synthesis, in particular to an aminopyrimidine compound, a pharmaceutical composition, application of the aminopyrimidine compound and an intermediate compound. The structural formula of the aminopyrimidine compound is shown as a formula (I), and the aminopyrimidine compound can effectively inhibit the activity of NUAKs kinase, and especially has a good inhibition effect on the activity of NUAK1 kinase and the activity of NNUAK2 kinase. Formula (I).

Description

Technical Field

[0001] This invention relates to the field of compound synthesis technology, specifically to an aminopyrimidine compound, a pharmaceutical composition and its application, and an intermediate compound. Background Technology

[0002] Protein kinases are a class of enzymes that phosphorylate protein molecules, transmitting cellular signals and regulating cellular functions and biological activities through phosphorylation reactions. They are widely distributed in cells and participate in a variety of important biological processes. Adenosine monophosphate activated protein kinase (AMPK) belongs to the serine / threonine protein kinase family. AMPK is a cellular energy sensor and regulator of metabolic homeostasis. Its structural feature is a heterotrimeric protein complex composed of a catalytically active α subunit (α1 / α2) and regulatory β (β1 / β2) and γ (γ1 / γ2 / γ3) subunits. It is a highly conserved intracellular ATP level sensor, a central mediator of cellular responses to energy stress and mitochondrial damage, and coordinates various functions of autophagy and mitochondrial biology. Its family has 12 members: MARK1, MARK2, MARK3, MARK4, MELK, QIK, QSK, SIK, BRSK1, BRSK2, NUAK1, and NUAK2.

[0003] The NUAK kinase family, including NUAK1 (ARK5) and NUAK2 (SNARK), belongs to the AMPK-associated protein kinase family and is a class of serine / threonine protein kinases. NUAK1 and NUAK2 share high homology, with 58% amino acid sequence identity and 82% kinase domain identity. NUAK1 is mainly expressed in the heart, kidney, liver, brain, skeletal muscle, and various tumors, and is associated with poor tumor prognosis. NUAK2 has some tissue specificity, being expressed in skin, kidney, uterus, ovary, and brain tissues. After activation, it mainly affects transcription in the cell nucleus. Both NUAK1 and NUAK2 proteins participate in the regulation of multiple intracellular signaling pathways and can be activated by LKB1 phosphorylation and calcium ion activation. In addition, AKT can activate NUAK1 by phosphorylating Ser at position 600; some growth factors, such as insulin-like growth factor 1 (IGF1) signaling pathway, can also increase NUAK1 activity; and NUAK1 activity is enhanced when skeletal muscle cells are in a contractile state. Both NUAK1 and NUAK2 can be activated by LKB1, but NUAK2 can be activated by autophosphorylation. NUAK2 can interact with the deubiquitinase USP9 on the X chromosome. In various cell types, stimuli such as hypoosmolarity, DNA damage, oxidation, and malnutrition can lead to the activation of NUAK2. Skeletal muscle cells can also lead to the activation of NUAK2. Studies have shown that NUAK1 / 2 plays an important role in the pathogenesis of metabolic diseases, tumors, diabetes, neurodegenerative diseases, and fibrotic diseases.

[0004] For example, existing technologies (Ge Yanfeng, Fan Xin, Zhuang Xiufen, et al. Dual role of NUAK family in tumors[J]. Journal of Medical Research, 2022, 51(11):13-16.) have shown that the relative expression level of NUAK1 in liver cancer tissue is significantly higher than that in adjacent normal liver tissue, and downregulating the expression of NUAK1 can inhibit the growth and migration of liver cancer cells.

[0005] Currently, inhibitors targeting NUAK1 / 2 lack efficacy and selectivity, and most are still in the preclinical research stage. Given the potential value of NUAK1 / 2 inhibitors in treating neurodegenerative diseases, diabetes, cancers (such as liver cancer, leukemia, and lymphoma), visceral fibrosis (such as cirrhosis, renal fibrosis, and pulmonary fibrosis), and skin fibrosis (such as scarring), there is an urgent need to develop a NUAK kinase inhibitor with good efficacy in inhibiting NUAK kinase.

[0006] Chinese Patent Publication No. CN116940565A discloses an isoindolinone aminopyrimidine compound as a NUAK kinase inhibitor, with the following structural formula:

[0007] Therefore, it is essential to develop an aminopyrimidine compound, a pharmaceutical composition and its application, and an intermediate compound that can solve the above-mentioned technical problems. Summary of the Invention

[0008] The purpose of this invention is to overcome the shortcomings of the prior art and provide an aminopyrimidine compound, pharmaceutical composition, application thereof, and intermediate compound with good inhibitory effect on NUAK kinase. This aminopyrimidine compound can effectively inhibit NUAK kinase activity, especially showing good inhibition of NUAK1 and NNUAK2 kinase activity.

[0009] This invention is achieved through the following technical solutions: The first aspect of this invention provides an aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs, wherein the structural formula of the aminopyrimidine compound is shown in formula (I):

[0010] Formula (I) in, Selected from , , , , , , , , , or ; Selected from or ; The wavy line indicates the connection point.

[0011] In one embodiment of the present invention, R1 is selected from -H and -C. 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2.

[0012] In one embodiment of the present invention, n is selected from an integer of 1 or 2, representing the... The ring can be replaced by one or two R1 groups.

[0013] In one embodiment of the present invention, X is selected from -NH-, -O-, or chemical bonds.

[0014] As one embodiment of the present invention, R a Selected from -H, -C 1-3Alkyl, -OC 1-3 Alkyl or -OCHF2, preferably -H or -OC 1-3 Alkyl or -OCHF2.

[0015] As one embodiment of the present invention, when R c When it is H, R b Selected from C 4-8 Heterocyclic alkanes or C 4-10 Bridged cyclic alkane group, the C 4-8 Heterocyclic alkanes or C 4-10 The bridged cycloalkane group optionally contains one, two, or three additional heteromers selected from N, O, and S, and is optionally separated by a group selected from H, halogen, carbonyl, -C. 1-3 Alkyl groups and -N(CH3)2 substituents are used for substitution.

[0016] Preferably, when R c When it is H, R b Selected from , , , , , , , or .

[0017] As one embodiment of the present invention, when R c When R is not H, b and R c The atoms therein form a ring C. The ring C is selected from C. 4-6 cycloalkyl or C 4-6 Contains an N-heterocyclic alkyl group; and the ring C is optionally separated by one or more elements selected from -H, halogen, -C. 1-3 Substitution of alkyl, -OH, and carbonyl groups.

[0018] Preferably, when R c When it is not H, Selected from or .

[0019] As one embodiment of the present invention, R d Selected from ring C. Wherein, ring C is selected from C. 4-6 cycloalkyl or C 4-6 Contains an N-heterocyclic alkyl group; and the ring C is optionally separated by one or more elements selected from -H, halogen, -C. 1-3 Substitution of alkyl, -OH, and carbonyl groups.

[0020] Preferably, R d Selected from or .

[0021] In one embodiment of the present invention, U and V are each independently selected from CH or N.

[0022] As one embodiment of the present invention Selected from , , , , , , , , , , or .

[0023] As one embodiment of the present invention, the structural formula of the aminopyrimidine compound is shown in formula (II): ; Equation (II) in, Selected from , , , , , , , , , , or ; R1 is selected from -H, -C 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2; n is an integer selected from 1 or 2; X is selected from -NH-, -O-, or chemical bonds; R b Selected from , , or .

[0024] Preferably, Selected from , , , , , , , , , , , or .

[0025] As one embodiment of the present invention, the structural formula of the aminopyrimidine compound is shown in formula (III): ; Equation (III) in, Selected from , or R1 is selected from -H, -C 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2; n is an integer selected from 1 or 2; X is selected from -NH- or chemical bond.

[0026] As one embodiment of the present invention, the structural formula of the aminopyrimidine compound is shown in formula (IV): ; Formula (IV) in, Selected from or ; R1 is selected from -H, -C 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2; W is selected from C or N; R2 is selected from H or -C 1-3 alkyl.

[0027] In one embodiment of the present invention, the aminopyrimidine compound is selected from any one of the following compounds (Table 1).

[0028] Table 1. Aminopyrimidine compounds

[0029] A second aspect of the present invention provides a pharmaceutical composition comprising the above-described aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, and pharmaceutically acceptable carriers.

[0030] The pharmaceutically acceptable carrier refers to a pharmaceutically acceptable carrier commonly used in the pharmaceutical field; the pharmaceutical composition can be prepared according to methods known in the art. Any dosage form suitable for human or animal use can be formulated by combining the aminopyrimidine compounds described in this invention or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, and one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants.

[0031] The aminopyrimidine compounds or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, or pharmaceutical compositions containing them described in this invention can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lungs and respiratory tract, skin, vagina, rectum, etc.

[0032] The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, and prodrugs in the pharmaceutical composition may be used alone or in combination with other drugs; the other drugs include chemotherapeutic drugs.

[0033] Preferably, the other chemotherapy drugs are selected from one or more of paclitaxel, carboplatin, cisplatin, capecitabine, irinotecan, and azacitidine.

[0034] A third aspect of the present invention provides a pharmaceutical composition comprising the above-described aminopyrimidine compounds or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, and other pharmaceuticals.

[0035] Preferably, the other drugs are selected from one or more of paclitaxel, carboplatin, cisplatin, capecitabine, irinotecan, and azacitidine.

[0036] The fourth aspect of the present invention provides the use of the above-mentioned aminopyrimidine compounds or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, or the above-mentioned pharmaceutical compositions in the preparation of a medicament for treating diseases caused by abnormal activation of NUAK kinase.

[0037] In one embodiment of the present invention, the NUAK kinase includes NUAK1 kinase and / or NUAK2 kinase.

[0038] As one embodiment of the present invention, the disease is selected from one or more of autoimmune diseases, inflammatory diseases, visceral fibrosis, skin fibrosis, and cancer.

[0039] Preferably, the cancer is selected from one or more of the following: gastrointestinal stromal tumors, esophageal cancer, gastric cancer, melanoma, glioma, glioblastoma, ovarian cancer, bladder cancer, pancreatic cancer, prostate cancer, lung cancer, breast cancer, kidney cancer, liver cancer, osteosarcoma, multiple myeloma, cervical cancer, bone metastases, papillary thyroid carcinoma, non-small cell lung cancer, and colorectal cancer.

[0040] Preferably, the disease is selected from one or more of the following: rheumatoid arthritis, inflammatory bowel disease, chronic obstructive pulmonary disease, asthma, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, liver fibrosis, renal fibrosis, sequelae of myocarditis, scleroderma, keloids, hypertrophic scars, traumatic and postoperative scars.

[0041] The fifth aspect of the present invention provides the use of the above-mentioned aminopyrimidine compounds or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, or the above-mentioned pharmaceutical compositions in the preparation of NUAK kinase inhibitors.

[0042] A sixth aspect of the present invention provides an intermediate compound comprising any one of the following compounds: , , , , , , , , , , , , , .

[0043] The beneficial effects of this invention are: This invention provides an aminopyrimidine compound that can effectively inhibit NUAKs kinase activity, especially the activity of NUAK1 kinase and NNUAK2 kinase. Detailed Implementation

[0044] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as a result. However, these embodiments are merely exemplary and do not constitute any limitation on the scope of the present invention. Those skilled in the art should understand that modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but all such modifications and substitutions fall within the protection scope of the present invention.

[0045] definition The various aspects and features of the present invention will be further described below.

[0046] All references cited in this invention are incorporated herein by reference in their entirety, and in the event of any inconsistency between the meanings expressed in these references and those expressed herein, the meanings expressed herein shall prevail. Furthermore, the various terms and phrases used in this invention have their general meanings known to those skilled in the art; however, this invention still intends to provide a more detailed explanation and interpretation of these terms and phrases. In the event of any inconsistency between the mentioned terms and their known meanings and those expressed herein, the meanings expressed herein shall prevail. The following are definitions of various terms used in this invention, and these definitions apply to all terms used throughout this specification, unless otherwise specified in the specific context.

[0047] The following provides definitions of various groups in the compounds of this invention, which, unless otherwise defined, are used consistently in the specification and claims.

[0048] As used herein, the term "cycloalkyl" whether used alone or as part of another group refers to a saturated carbocyclic group containing 3 to 8 carbon atoms and one or more rings. The possible number of carbon atoms in the cycloalkyl group is indicated by the numerical prefix "C". n1-n2 "instruct.

[0049] For example, term C 4-8 Cycloalkyl refers to a cycloalkyl group having 4, 5, 6, 7, or 8 carbon atoms; the term "alkyl" refers to an alkyl group having a specified number of carbon atoms, which can be straight-chain or branched, such as the "C" mentioned. 1-3 When "alkyl" is mentioned, it includes methyl, ethyl, propyl, and isopropyl; for example, when mentioning "C 4-8 When "heterocyclic group" is used, it refers to a non-aromatic heterocycle with 4, 5, 6, 7 or 8 carbon atoms, including saturated heterocycles and non-aromatic unsaturated heterocycles with double bonds. For example, when "bridged (condensed) heterocyclic group" is mentioned, it also includes saturated bridged (condensed) heterocycles and non-aromatic unsaturated bridged (condensed) heterocycles with double bonds.

[0050] Example 1: 3-Chloro-N6-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-N2-(1H-pyrrolo[2,3-b]pyridin-5-yl)pyridine-2,6-diamine (B001)

[0051] Step 1: Synthesis of intermediates 1-3: 2,4,5-Trichloropyrimidine (415 mg, 2.26 mmol) was placed in a round-bottom flask, and 10 mL of ethanol was added to fully dissolve the raw material. Then, 1H-pyrrolo[2,3-b]pyridine-5-amine (300 mg, 2.26 mmol) and sodium carbonate (480 mg, 4.52 mmol) were added, and the mixture was heated to 80°C. o Stirred overnight at C. After the reaction was complete, the mixture was extracted three times with ethyl acetate. After removing the solvent under vacuum, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography (PE:EA) = 3:1, yielding 400 mg of a white solid (1-3). ESI-MS: m / z = 280 [M+H] + .

[0052] Step 2: Synthesis of intermediates 1-5: Raw materials 1-4 (4 g, 23.4 mmol) were dissolved in 40 mL of DMSO. N-methylpiperazine (2.8 g, 28.08 mmol) was added dropwise at room temperature, followed by potassium carbonate (9.7 g, 70.2 mmol). The mixture was heated to 85 °C and stirred overnight. After cooling to room temperature, water and EA were added, and the mixture was extracted three times with EA. The organic phase was dried over anhydrous sodium sulfate and concentrated to give 6 g of intermediate 1-5. ESI-MS: m / z = 252 [M+H] + .

[0053] Step 3: Synthesis of intermediates 1-6: Intermediates 1-5 (502 mg, 2 mmol) were placed in a round-bottom flask, and 10 mL of methanol was added to fully dissolve the starting material. Then, 100 mg of palladium on carbon was added, and the mixture was stirred overnight under hydrogen atmosphere. After the reaction was complete, the mixture was cooled to room temperature, filtered through a membrane filter, and concentrated to give 400 mg of the product as a white solid (1-6). ESI-MS: m / z = 222 [M+H] + .

[0054] Step 4: Synthesis of compound B001: Intermediates 1-3 (280 mg, 1 mmol) and 1-6 (332 mg, 1.5 mmol) were dissolved in 20 mL of 1,4-dioxane solution, and 0.5 mL of 1,4-dioxane chloride (2 mmol) was added. The reaction was carried out overnight at 120 °C under nitrogen protection. After the reaction was completed, the mixture was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography (DCM:MeOH) = 12:1 to give 200 mg of compound B001.

[0055] 1 H NMR (400 MHz, DMSO-d 6) δ 11.55 (s, 1H), 8.80 (s, 1H), 8.22 (d, J =2.4 Hz, 1H), 8.09 (d, J = 2.3 Hz, 1H), 7.98 (s, 1H), 7.60 (s, 1H), 7.45 – 7.37(m, 2H), 6.52 (d, J = 2.5 Hz, 1H), 6.36 (dd, J = 3.4, 1.9 Hz, 1H), 6.11 (d, J = 8.7Hz, 1H), 3.71 (s, 3H), 3.01 (t, J = 5.0 Hz, 4H), 2.46 – 2.39 (m, 6H), 2.20 (s, 3H). ESI-MS: m / z = 464 [M+H] + .

[0056] Example 2: 2-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)tetrahydrobenzo[b]thiophene-3-carboxynitrile (B002)

[0057] Step 1: Synthesis of intermediate 2-2: The synthesis steps were the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 2-1 to obtain intermediate 2-2. ESI-MS: m / z = 325 [M + H] + .

[0058] Step 2: Synthesis of compound B002: Intermediate 2-2 (115 mg, 0.35 mmol), 2-methoxy-4-(4-methylpiperazin-1-yl)aniline (117.8 mg, 0.53 mmol), and TsOH (66.6 mg, 0.35 mmol) were added to 2 mL of IPA and heated to 100 °C. o The reaction was carried out at C for 17 hours. After the reaction was completed, the mixture was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and separated by column chromatography (DCM: MeOH) = 30:1 to give 64 mg of compound B002.

[0059] 1 H NMR (400 MHz, Chloroform-d ) δ 8.09 (s, 1H), 7.94 (s, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.11 (s, 1H), 6.53 (d, J = 1.0 Hz, 2H), 3.86 (s, 3H), 3.22 –3.14 (m, 4H), 2.63 – 2.54 (m, 8H), 2.36 (s, 3H), 1.88 – 1.75 (m, 4H). ESI-MS; m / z = 510 [M + H] + .

[0060] Example 3: 5-(5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylonitrile (B003)

[0061] Step 1: Synthesis of intermediate 3-2: Intermediate 3-1 (500 mg, 2.85 mmol) was added to 5 mL of DCM. o Add TEA (576.8 mg, 5.7 mmol) at C, stir for ten minutes, then add Boc2O (621.2 mg, 2.85 mmol) dissolved in 5 mL of DCM. o The reaction was carried out at C for 3 hours. After the reaction was complete, the reaction system was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 5:1) to give 580 mg of the product as a white solid (3-2), ESI-MS; m / z = 240 [M + H]. + .

[0062] Step 2: Synthesis of intermediate 3-3: Intermediate 3-2 (580 mg, 2.85 mmol) was added to 5 mL of DCM, followed by NBS (536.2 mg, 3 mmol). 25 mL of DCM was then added. o The reaction was carried out at C for 3 hours. After the reaction was complete, the reaction system was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 5:1) to give 575 mg of white solid (3-3), ESI-MS; m / z = 318 [M + H]. + .

[0063] Step 3: Synthesis of intermediates 3-4: Intermediate 3-3 (390 mg, 1.43 mmol), Zn(CN)2 (419.7 mg, 3.58 mmol), and Pd(dppf)Cl2 (102.4 mg, 0.14 mmol) were added to 4 mL of DMF and sealed in a container for 150 minutes. o The reaction was carried out at C for 4 hours. After the reaction was complete, the reaction system was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 4:1) to give 210 mg of a pale yellow oily liquid (3-4), ESI-MS; m / z = 165 [M + H] + .

[0064] Step 4: Synthesis of intermediates 3-5: The synthesis steps are the same as in Step 1 of Example 2, except that intermediate 2-1 is replaced with intermediate 3-4 to obtain intermediate 3-5. ESI-MS: m / z = 311 [M + H] + .

[0065] Step 5: Synthesis of compound B003: The synthesis of intermediates 3-5 (140 mg, 0.45 mmol), intermediates 1-6 (149.4 mg, 0.68 mmol), cesium carbonate (293.2 mg, 0.9 mmol), Pd(OAc)2 (6.9 mg, 0.045 mmol), and BINAP (28 mg, 0.045 mmol) were added to 2 mL of dioxane, purged with nitrogen, and 110 o The reaction was carried out at C for 16 hours. After the reaction was complete, the reaction system was added to 10 mL of water, extracted with ethyl acetate (20 mL x 4), the organic phases were combined, washed with saturated brine (25 mL x 2), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (EA : MeOH = 10 : 1), and then slurried in a mixed solvent of n-hexane and dichloromethane (4:1) to obtain 80 mg of compound B003 as a white solid. 1 H NMR (400 MHz, DMSO- d 6)δ 8.04 (s, 1H), 7.80 (s, 1H), 7.66 (d, J = 8.7 Hz, 1H), 7.54 (s, 1H), 6.61 (d, J = 2.5 Hz, 1H), 6.47 (dd, J = 8.9, 2.5 Hz, 1H), 4.59 (s, 2H), 3.86 (t, J= 5.6 Hz,2H), 3.80 (s, 3H), 3.11 (t, J = 4.9 Hz, 4H), 2.95 (t, J = 5.6 Hz, 2H), 2.46 (t, J =5.0 Hz, 4H), 2.22 (s, 3H). ESI-MS; m / z = 496 [M + H] + .

[0066] Example 4: 5-(5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxamide (B004)

[0067] Step 1: Synthesis of compound B004: Compound B003 (40 mg, 0.081 mmol), triphenylphosphine (2.6 mg, 0.01 mmol), palladium acetate (1.8 mg, 0.008 mmol), and acetaldehyde oxime (9.6 mg, 0.16 mmol) were added to 5 mL of a mixed solvent of ethanol / water (4:1). o The reaction was carried out at C for 4 h. After the reaction was complete, the reaction mixture was added to 5 mL of water, extracted with ethyl acetate (10 mL x 5), the extracts were combined, washed with saturated brine (15 mL x 2), dried over sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by TLC (DCM:MeOH = 10:1) to obtain compound B004. 1 H NMR (400 MHz, DMSO- d 6) δ 8.04 (s, 1H), 7.80 (s, 1H), 7.66 (d, J = 8.7 Hz, 1H), 7.54 (s, 1H), 6.61 (d, J = 2.5 Hz, 1H), 6.47 (dd, J = 8.9, 2.5 Hz, 1H), 4.59 (s, 2H), 3.86 (t, J = 5.6 Hz, 2H), 3.80 (s, 3H), 3.11(t, J = 4.9 Hz, 4H), 2.95 (t, J = 5.6 Hz, 2H), 2.46 (t, J= 5.0 Hz, 4H), 2.22 (s,3H). ESI-MS; m / z = 514 [M + H] + .

[0068] Example 5: N4-(benzo[b]thiophene-5-yl)-5-chloro-N2-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyrimidine-2,4-diamine (B005)

[0069] Step 1: Synthesis of intermediate 5-2: The synthesis steps are the same as in Step 1 of Example 1, except that intermediate 5-1 was used instead of intermediate 1-2 to obtain intermediate 5-2. ESI-MS: m / z = 296 [M+H] + Step 2: Synthesis of compound B005: The synthesis procedure was the same as step 2 of Example 2, except that intermediate 2-2 was replaced with intermediate 5-2 to obtain compound B005. ESI-MS: m / z = 481 [M+H] + .

[0070] Example 6: 5-(5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-4,5,6,7-tetrahydrothiazo[5,4-c]pyridine-2-amine (B006)

[0071] Step 1: Synthesis of intermediate 6-2: Intermediate 6-1 (450 mg, 1.76 mmol) and fluorenyl chloroformate (501 mg, 1.94 mmol) were added to 10 mL of THF. Sodium bicarbonate (296 mg, 3.52 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 5 h. Water (30 mL) was added to the reaction mixture, and the mixture was extracted with EtOAc (3 × 30 mL). The organic phase was washed with saturated brine (3 × 30 mL), dried over anhydrous sodium sulfate, and purified by column chromatography (PE:EtOAc = 3:1) to give 534 mg of intermediate 6-2. ESI-MS: m / z = 478 [M+H] + .

[0072] Step 2: Synthesis of intermediate 6-3: Intermediate 6-2 (530 mg, 1.11 mmol) was added to 3 mL of DCM, and the mixture was placed in an ice bath at 0°C. Then, 2 mL of HCl solution (4 M in 1,4-Dioxane, 8 mmol) was added dropwise to the reaction mixture, and the mixture was brought back to room temperature and stirred for 6 h. After the reaction was complete, the solvent was removed by rotary evaporation to obtain intermediate 6-3. ESI-MS: m / z = 378 [M+H] + .

[0073] Step 3: Synthesis of intermediate 6-4: The synthesis steps are the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 6-3 to obtain intermediate 6-4. ESI-MS: m / z = 524 [M+H] + .

[0074] Step 4: Synthesis of intermediates 6-5: The synthesis steps are the same as in step 2 of Example 2, except that intermediate 2-2 is replaced with intermediate 6-4 to obtain intermediate 6-5. ESI-MS: m / z=709 [M+H] + .

[0075] Step 5: Synthesis of compound B006: Intermediate 6-5 was dissolved in 10 mL of methanol, and 0.66 mL of piperidine was added. The mixture was reacted at room temperature for 18 hours. The reaction solution was then concentrated, and the residue was purified by column chromatography (ethyl acetate: methanol = 10:1) to give compound B006. ESI-MS: m / z = 487 [M+H] + .

[0076] Example 7: N2-(2-(difluoromethoxy)-4-(4-methylpiperazin-1-yl)phenyl)-N4-(5-methyl-4,5,6,7-tetrahydrothiazo[5,4-c]pyridin-2-yl)pyrimidine-2,4-diamine (B007)

[0077] Step 1: Synthesis of intermediate 7-3: Intermediate 7-1 (785 mg, 5 mmol) was dissolved in 30 mL of anhydrous THF. Sodium hydride was added in portions at -15 °C, and the mixture was stirred for 30 min. Then, water (6.3 g, 350 mmol) was slowly added dropwise at -7 °C, and the mixture was stirred for 30 min. Intermediate 7-2 (2.67 g, 10 mmol) was slowly added dropwise at -7 °C, and the mixture was allowed to return to room temperature and stirred for 1 hour. EA was added to the reaction mixture, and the solution was dried over anhydrous sodium sulfate. After filtration and washing several times with EA, the product was concentrated into a white solid of 1.65 g (7-3). ESI-MS: m / z = 208 [M+H]+ .

[0078] Step 2: Synthesis of intermediate 7-4: The synthesis procedure follows the same steps as in Step 2 of Example 1, except that intermediate 1-4 is replaced with intermediate 7-3 to obtain intermediate 7-4. ESI-MS: m / z = 288 [M+H] + .

[0079] Step 3: Synthesis of intermediate 7-5: The synthesis procedure follows the same steps as in Step 3 of Example 1, except that intermediate 1-5 is replaced with intermediate 7-4 to obtain intermediate 7-5. ESI-MS: m / z = 258 [M+H] + .

[0080] Step 4: Synthesis of intermediate 7-7: The synthesis procedure follows the same steps as in Example 1, step 1, except that intermediate 7-6 is used instead of intermediate 1-2 to obtain intermediate 7-7. ESI-MS: m / z = 282 [M+H] + .

[0081] Step 5: Synthesis of compound B007: The synthesis procedure was the same as step 2 in Example 2, except that intermediate 2-2 was replaced with intermediate 7-7, and intermediate 1-6 was replaced with intermediate 7-5, yielding compound B007. ESI-MS: m / z = 503 [M+H] + .

[0082] Example 8: 2-((5-chloro-2-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)-5,5-dimethyl-5,6-dihydrobenzo[d]thiazolyl-7(4H)-one (B008)

[0083] Step 1: Synthesis of intermediate 8-2: The synthesis steps are the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 8-1 to obtain intermediate 8-2. ESI-MS: m / z = 343 [M+H] + .

[0084] Step 2: Synthesis of compound B008: The synthesis procedure was the same as step 2 of Example 2, except that intermediate 2-2 was replaced with intermediate 8-2 to obtain compound B008. ESI-MS: m / z = 528 [M+H] + .

[0085] Example 9: N4-(1H-benzo[d]imidazol-5-yl)-5-chloro-N2-(2-methoxy-4-morpholinophenyl)pyrimidine-2,4-diamine (B009)

[0086] Step 1: Synthesis of intermediate 9-2: The synthesis steps were the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 9-1 to obtain intermediate 9-2. ESI-MS: m / z = 280 [M + H] + .

[0087] Step 2: Synthesis of intermediate 9-3: The synthesis procedure was the same as step 2 in Example 1, except that N-methylpiperazine was replaced with morpholine to obtain intermediate 9-3. ESI-MS: m / z = 239 [M + H] + .

[0088] Step 3: Synthesis of intermediate 9-4: The synthesis steps were the same as in step 3 of Example 1, except that intermediate 9-3 was used instead of intermediate 1-5 to obtain intermediate 9-4. ESI-MS: m / z = 209 [M + H] + .

[0089] Step 4: Synthesis of compound B009: The synthesis procedure was the same as step 4 of Example 1, except that intermediate 1-6 was replaced with intermediate 9-4, and intermediate 1-3 was replaced with intermediate 9-2, to obtain compound B009. ESI-MS: m / z = 452 [M + H] + .

[0090] Example 10: 4-(4-((5-chloro-4-(5,6-dihydroimidazol[1,5-a]pyrazin-7(8H)-yl)pyrimidin-2-yl)amino)-3-methoxyphenyl)thiomorpholine 1,1-dioxide (BO10)

[0091] Step 1: Synthesis of intermediate 10-2: The synthesis steps were the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 10-1 to obtain intermediate 10-2. ESI-MS: m / z = 270 [M + H] + .

[0092] Step 2: Synthesis of intermediate 10-3: The synthesis procedure was the same as step 2 in Example 1, except that N-methylpiperazine was replaced with thiomorpholine to obtain intermediate 10⁻³. ESI-MS: m / z = 255 [M + H] + .

[0093] Step 3: Synthesis of intermediate 10-4: The synthesis procedure was the same as step 3 in Example 1, except that intermediate 1-5 was replaced with intermediate 10-3 to obtain intermediate 10-4. ESI-MS: m / z = 225 [M + H] + .

[0094] Step 4: Synthesis of intermediate 10-5: The synthesis steps were the same as in step 4 of Example 1, except that intermediate 1-6 was replaced with intermediate 10-4, and intermediate 1-3 was replaced with intermediate 10-2, resulting in intermediate 10-5. ESI-MS: m / z = 458 [M + H] + .

[0095] Step 5: Synthesis of compound B010: Intermediate 10⁻⁵ (150 mg, 0.33 mmol) and m-chloroperoxybenzoic acid (113.90 mg, 0.66 mmol) were added sequentially to a round-bottom flask, followed by 3 mL of dichloromethane. The mixture was stirred at room temperature for approximately 4 h. After the reaction was complete, 5 mL of saturated sodium thiosulfate solution was added to quench the reaction mixture, and the mixture was extracted three times with dichloromethane. The organic phase was washed with saturated brine (3 × 15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by thin-layer chromatography (DCM:MeOH = 12:1) to yield 15 mg of product BO10. 1 H NMR (400 MHz, DMSO- d 6) δ 8.16 (s, 1H), 8.14 (d, J = 8.9 Hz,1H), 8.07 (s, 1H), 7.90 (d, J = 2.4 Hz, 1H), 7.60 – 7.56 (m, 2H), 6.78 (s, 1H), 4.79 (s, 2H), 4.48 – 4.39 (m, 2H), 4.16 (t, 2H), 3.98 (t, J = 5.5 Hz, 2H), 3.89(s, 3H), 3.76 – 3.66 (m, 2H), 3.06 – 2.94 (m, 4H). ESI-MS: m / z =490 [M+H] + .

[0096] Example 11: 3-(4-((5-chloro-4-(4,7-dihydrothieno[2,3-c]pyridin-6(5H)-yl)pyrimidin-2-yl)amino)-3-methoxyphenyl)-N,N-dimethyl-3-azabicyclo[3.1.0]hexyl-6-amine (B011)

[0097] Step 1: Synthesis of intermediate 11-2: The synthesis steps were the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 11-1 to obtain intermediate 11-2. ESI-MS: m / z = 286 [M + H] + .

[0098] Step 2: Synthesis of intermediate 11-5: Intermediate 11-3 (2 g, 11.8 mmol) was added to 10 mL of THF, followed by Ti(OiPr)4 (4.04 g, 14.2 mmol). While stirring, MeMgCl (1 N in THF; 14.2 mL, 14.2 mmol) was slowly added dropwise, followed by DMF (1.04 g, 14.2 mmol). Then, cHexMgBr (1 N in THF; 23.6 mL, 23.6 mmol) was slowly added. The mixture was heated to 80°C. o Reflux at C for 1 hour. After the reaction is complete, evaporate the reaction system to dryness to obtain crude intermediate 11-5. ESI-MS; m / z = 227 [M+ H] + .

[0099] Step 3: Synthesis of intermediate 11-6: The synthesis steps were the same as in step 2 of Example 6, except that intermediate 6-2 was replaced with intermediate 11-5 to obtain intermediate 11-6. ESI-MS: m / z = 127 [M + H] + .

[0100] Step 4: Synthesis of intermediate 11-7: The synthesis procedure was the same as step 2 of Example 1, except that intermediate 11-6 was used instead of N-methylpiperazine to obtain intermediate 11-7. ESI-MS: m / z = 278 [M + H] + .

[0101] Step 5: Synthesis of intermediates 11-8: The synthesis steps are the same as in step 3 of Example 1, except that intermediate 1-5 is replaced with intermediate 11-7 to obtain intermediate 11-8. ESI-MS: m / z = 248 [M + H] + .

[0102] Step 6: Synthesis of compound B011: The synthesis steps are the same as in step 4 of Example 1, with intermediate 1-6 replaced by intermediate 1-8 and intermediate 1-3 replaced by intermediate 11-2, to obtain compound B011. 1 H NMR (400 MHz, DMSO- d 6) δ 7.98 (s, 1H), 7.73 (s, 1H),7.48 (d, J = 8.6 Hz, 1H), 7.36 (d, J = 5.0 Hz, 1H), 6.86 (d, J = 5.1 Hz, 1H), 6.17(d, J = 2.5 Hz, 1H), 6.06 (dd, J = 8.7, 2.5 Hz, 1H), 4.73 (s, 2H), 3.83 (t, J = 5.7Hz, 2H), 3.75 (s, 3H), 3.47 (d, J = 9.3 Hz, 2H), 3.16 (dt, J = 9.1, 2.2 Hz, 2H), 2.78 (t, J = 5.7 Hz, 2H), 2.25 (s, 6H), 1.72 – 1.68 (m, 2H), 1.45 (t, J = 2.1 Hz,1H). ESI-MS; m / z = 497 [M + H] + .

[0103] Example 12: 5-Chloro-N2-(2-methyl-1,2,3,4-tetrahydroisoquinoline-6-yl)-N4-(1H-pyrrolo[2,3-b]pyridin-5-yl)pyrimidine-2,4-diamine (B012)

[0104] Step 1: Synthesis of Intermediate 12-2: 6-Nitro-1,2,3,4-Tetrahydroisoquinoline hydrochloride (500 mg, 2.33 mmol) was added to a round-bottom flask, followed by 5 mL of dichloromethane, then TEA (356.5 μL, 2.56 mmol). After stirring for 5 min, paraformaldehyde (280 mg, 9.32 mmol) was added. The mixture was cooled to approximately 0°C in an ice bath, and then sodium triacetoxyborohydride (1.03 g, 4.89 mmol) was added. The mixture was stirred at 0°C for another 30 min, and the reaction was allowed to proceed for approximately 16 h at room temperature. After the reaction was complete, the reaction solution was washed three times with saturated sodium carbonate, and the organic layer was recovered. The organic layer was dried over anhydrous sodium sulfate and evaporated under reduced pressure to give 364.8 mg (12-2) of a pale yellow solid. ESI-MS: m / z = 193 [M+H] + .

[0105] Step 2: Synthesis of intermediate 12-3: Intermediate 12-2 (450 mg, 2.30 mmol) was added to a round-bottom flask, followed by 5 mL of anhydrous methanol, and then Raney nickel (2.3 mL). The mixture was purged three times with hydrogen, and the reaction was stirred overnight at room temperature under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered through diatomaceous earth, the organic layer was recovered, and evaporated under reduced pressure to give 360 ​​mg (12-3) of a brown solid. ESI-MS: m / z = 163 [M+H] + .

[0106] Step 3: Synthesis of compound B012: The synthesis steps are the same as in step 2 of Example 2, except that intermediate 2-2 is replaced with intermediate 12-3 to obtain compound B012. 1 HNMR (400 MHz, DMSO- d 6) δ 11.71 (s, 1H), 9.14 (s, 1H), 8.95 (s, 1H), 8.27 (d, J = 2.3 Hz, 1H), 8.08 (s, 1H), 8.04 (d, J = 2.3 Hz, 1H), 7.49 (t, J = 3.0 Hz, 1H),7.26 (s, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.68 (d, J = 8.4 Hz, 1H), 6.43 (dd, J =3.2, 1.8 Hz, 1H), 3.29 (s, 2H), 2.39 (t, J= 5.9 Hz, 2H), 2.25 (s, 3H), 2.21(s, 2H). ESI-MS: m / z = 406 [M+H] + .

[0107] Example 13: N-(5-chloro-4-(6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)pyrimidin-2-yl)-2-methyl-1,2,3,4-tetrahydroisoquinoline-6-amine (B013)

[0108] Step 1: Synthesis of Intermediate 13-2: The synthesis steps were the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 13-1 to obtain intermediate 13-2. ESI-MS: m / z = 286 [M + H] + .

[0109] Step 2: Synthesis of compound B013: The synthesis steps are the same as in step 2 of Example 2, except that intermediate 13-2 is used instead of intermediate 2-2, and intermediate 12-3 is used instead of intermediate 1-6 to obtain compound B013. 1 H NMR (400 MHz, Chloroform- d ) δ 7.98 (s, 1H), 7.41 (s, 1H), 7.23 (dd, J = 8.3, 2.3 Hz, 1H), 7.13 (d, J = 5.2 Hz, 1H), 6.96 (d, J = 8.8Hz, 2H), 6.81 (d, J = 5.1 Hz, 1H), 4.74 (t, J = 1.7 Hz, 2H), 3.98 (t, J = 5.6 Hz, 2H), 3.57 (s, 2H), 3.09 – 3.03 (m, 2H), 2.93 (t, J = 6.0 Hz, 2H), 2.71 (t, J =5.9 Hz, 2H), 2.47 (s, 3H). ESI-MS: m / z =412 [M+H] + .

[0110] Example 14: N-(5-chloro-4-(5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)pyrimidin-2-yl)-2-methyl-1,2,3,4-tetrahydroisoquinoline-6-amine (B014)

[0111] Step 1: Synthesis of compound B014: The synthesis procedure was the same as step 2 of Example 2, except that intermediate 2-2 was replaced with intermediate 10-2, and intermediate 1-6 was replaced with intermediate 12-3, yielding compound B014. ESI-MS: m / z = 396 [M+H] + .

[0112] Example 15: 5-Chloro-4-(4,7-dihydrothieno[2,3-c]pyridin-6(5H)-yl)-N-(5-(4-methylpiperazin-1-yl)pyrazin-2-yl)pyrimidin-2-amine (B015)

[0113] Step 1: Synthesis of Intermediate 15-2: The synthesis steps are the same as in step 1 of Example 3, except that intermediate 3-1 was replaced with intermediate 15-1 to obtain intermediate 15-2. ESI-MS: m / z = 274 [M+H] + .

[0114] Step 2: Synthesis of Intermediate 15-3: Intermediate 15-2 (365 mg, 1.34 mmol) and N-methylpiperazine (201 mg, 2.01 mmol) were dissolved in 12 mL of toluene. Davephos (58 mg, 0.15 mmol), palladium acetate (15.7 mg, 0.15 mmol), and t-BuONa (386.5 mg, 4.02 mmol) were added, and the mixture was purged with nitrogen. 100 o The reaction was carried out overnight at C, filtered through diatomaceous earth, concentrated, and then separated by column chromatography (PE:EA = 10:1) to give 24 mg (15-3) of white solid product. ESI-MS: m / z = 294 [M+H] + .

[0115] Step 3: Synthesis of intermediate 15-4: The synthesis procedure follows the same steps as in Step 2 of Example 6, except that intermediate 6-2 is replaced with intermediate 15-3 to obtain intermediate 15-4. ESI-MS: m / z = 194 [M+H] + .

[0116] Step 4: Synthesis of compound B015: The synthesis steps are the same as in step 2 of Example 2, except that intermediate 11-2 is used instead of intermediate 2-2, and intermediate 15-4 is used instead of intermediate 1-6 to obtain compound B015. 1 H NMR (400 MHz, Chloroform- d ) δ 9.07 (d, J = 1.5 Hz,1H), 8.19 (s, 1H), 8.11 (s, 1H), 7.88 (d, J = 1.6 Hz, 1H), 7.16 (d, J = 5.0 Hz, 1H), 6.83 (d, J = 5.1 Hz, 1H), 4.88 (s, 2H), 3.96 (t, J = 5.6 Hz, 2H), 3.55 (t, J =5.1 Hz, 4H), 2.94 (t, J = 5.6 Hz, 2H), 2.57 (t, J = 5.1 Hz, 4H), 2.37 (s, 3H).ESI-MS: m / z = 443 [M+H] + .

[0117] Example 16: 5-Chloro-4-(6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyrimidin-2-amine (B016)

[0118] Step 1: Synthesis of Intermediate 16-2: Intermediate 16-1 (1.1 g, 5.5 mmol), 4-nitropyrazole (635.5 mg, 5 mmol), and triphenylphosphine (1.97 g, 7.5 mmol) were dissolved in 25 mL of anhydrous THF. o Diisopropylazo-1,2-dicarboxylic acid ester was added dropwise at C, and the reaction was allowed to proceed overnight. The reaction solvent was evaporated to dryness, and water and ethyl acetate were added. The mixture was extracted three times with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and separated by column chromatography (PE:EA = 2:1). ESI-MS: m / z = 297 [M+H] + .

[0119] Step 2: Synthesis of intermediate 16-3: The synthesis procedure follows the same steps as in Step 2 of Example 12, except that intermediate 12-2 is replaced with intermediate 16-2 to obtain intermediate 16-3. ESI-MS: m / z = 267 [M+H] + .

[0120] Step 3: Synthesis of intermediate 16-4: Intermediate 13-2 (160.0 mg, 0.556 mmol) and intermediate 16-3 (178.7 mg, 0.671 mmol) were dissolved in 5 mL of 1,4-dioxane. Cs₂CO₃ (546.5 mg, 1.68 mmol), X-Phos (26.65 mg, 0.056 mmol), and Pd₂(dba)₃ (51.2 mg, 0.056 mmol) were added sequentially. Under N₂ protection, the mixture was heated to 120 °C and reacted for 4 h. After the reaction was complete, the mixture was extracted three times with EA, washed successively with water and saturated brine, and the organic phase was dried, filtered, and evaporated to dryness. Separation was performed by column chromatography (PE:EA = 1:1) to give 12 mg (16-4) of the pale yellow solid. ESI-MS: m / z = 516 [M+H] + .

[0121] Step 4: Synthesis of compound B016: The synthesis procedure was the same as step 2 in Example 6, except that intermediate 6-2 was replaced with intermediate 16-4 to obtain compound B016. ESI-MS: m / z = 416 [M+H] + .

[0122] Example 17: 5-Chloro-4-(6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyrimidin-2-amine (B017)

[0123] Step 1: Synthesis of Intermediate 17-2: The synthesis steps are the same as in step 1 of Example 16, except that intermediate 16-1 was replaced with intermediate 17-1 to obtain intermediate 17-2. ESI-MS: m / z = 211 [M+H] + .

[0124] Step 2: Synthesis of intermediate 17-3: The synthesis steps are the same as in step 2 of Example 16, except that intermediate 16-2 was replaced with intermediate 17-2 to obtain intermediate 17-3. ESI-MS: m / z = 181 [M+H] + .

[0125] Step 3: Synthesis of compound B017: The synthesis steps are the same as in step 3 of Example 16, except that intermediate 16-3 is replaced with intermediate 17-3 to obtain compound B017. 1 H NMR (400 MHz, Chloroform- d ) δ 7.96 (s, 2H), 7.76 (s, 2H), 7.52 (s, 2H), 7.25 (s, 1H), 7.14 (d, J = 5.0 Hz, 2H), 6.83 – 6.79 (m, 2H), 5.28 (s, 7H), 4.84(s, 4H), 4.14 (s, 2H), 3.94 (t, J = 5.6 Hz, 4H), 3.07 (d, J = 8.4 Hz, 4H), 2.89(d, J = 5.4 Hz, 4H), 2.40 (s, 6H), 2.36 – 2.22 (m, 7H), 2.13 (s, 4H), 2.03 (s,1H), 1.24 (s, 15H). ESI-MS: m / z =430 [M+H] + .

[0126] Example 18: 5-chloro-4-(5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyrimidin-2-amine (B018)

[0127] Step 1: Synthesis of compound B018: The synthesis procedure was the same as step 3 of Example 16, except that intermediate 16-3 was replaced with intermediate 17-3 and intermediate 13-2 was replaced with intermediate 10-2, yielding compound B018. ESI-MS: m / z = 414 [M+H] + .

[0128] Example 19: 4-((1H-indazol-6-yl)oxy)-5-chloro-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyrimidin-2-amine (B019)

[0129] Step 1: Synthesis of intermediate 19-2: The synthesis steps were the same as in Step 1 of Example 1, except that intermediate 1-2 was replaced with intermediate 19-1 to obtain intermediate 19-2. ESI-MS: m / z = 281 [M + H] + .

[0130] Step 2: Synthesis of compound B019: The synthesis steps are the same as in step 2 of Example 2, except that intermediate 2-2 is replaced with intermediate 19-2 to obtain compound B019. 1 HNMR (400 MHz, DMSO- d 6) δ 8.30 (s, 1H), 8.09 (d, J = 1.1 Hz, 1H), 8.04 (s, 1H), 7.79 (d, J = 8.7 Hz, 1H), 7.38 – 7.35 (m, 1H), 7.04 (d, J = 8.4 Hz, 1H), 6.95 (dd, J = 8.6, 2.0 Hz, 1H), 6.43 (d, J = 2.7 Hz, 1H), 3.66 (s, 3H), 2.94 (t, J = 4.9Hz, 4H), 2.38 (t, J = 5.0 Hz, 4H), 2.18 (s, 3H). ESI-MS; m / z =466 [M + H] + .

[0131] Example 20: In vitro NUAK1 / 2 kinase activity inhibition experiment Experimental objective: To detect the in vitro inhibitory activity of the test compounds on the NUAK1 and NUAK2 kinases using the ADP-Glo ​​method.

[0132] Experimental methods: ADP-Glo™ is a luminescent kinase assay that measures kinase activity by detecting the amount of ADP formed during a kinase reaction. In this assay, any remaining ATP from the kinase reaction is first consumed by the ADP-Glo ​​reagent; subsequently, the ADP generated is reduced back to ATP by the kinase assay reagent; then, the ATP reacts with luciferin under the action of Ultra-Glo™ luciferase to produce light. The luminescence signal is measured using an Envision plate reader, and the luminescence signal is positively correlated with kinase activity. In this specific study, the kinase sources were: NUAK1 (manufacturer: Carna, catalog number: 02-126); NUAK2 (manufacturer: Carna, catalog number: 02-127); the assay kit was: ADP-Glo™ Kinase Assay (manufacturer: Promega, catalog number: V9103).

[0133] Reaction system: 1. Prepare kinase reaction buffer: 50 mM Hepes, 10 mM MgCl2, 0.01% Brij-35, 1 mM EGTA, 2 mM DTT, H2O.

[0134] 2. Compound preparation: All compounds were dissolved and diluted with 100% DMSO to obtain compound dilution solutions.

[0135] 3. Prepare 2× ATP / substrate (KKKVSRSGLYRSPSMPENLNRPR) solution and 2× kinase solution using kinase reaction buffer. The final concentration of substrate is 0.5 mM, the final concentration of ATP is 125 μM, and the final concentration of kinase is 20 ng / 50 μL.

[0136] 4. Using an Echo 655, transfer 40 nL of the compound dilution to a 384 assay plate as the compound well; centrifuge, add 2 μL of 2× kinase solution to the 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 10 minutes. Transfer 40 nL of 100% DMSO to the 384 assay plate as the DMSO well. Wells without any reagents are used as blank wells. No further processing is performed on the DMSO wells and blank wells. Each well is prepared in triplicate.

[0137] 5. Add 2 μL of 2× substrate / ATP solution to the 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 60 minutes.

[0138] 6. Transfer 4 μL of ADP-Glo ​​to a 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes to terminate the reaction and obtain the detection solution.

[0139] 7. Transfer 8 μL of the detection solution obtained in step 6 to a 384 plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes.

[0140] 8. Use a multi-functional microplate reader to read the luminescent signal.

[0141] Data processing: The inhibition rate is calculated as follows: Inhibition rate (%) = 100% - (ave compound wells - ave blank wells -) / (ave DMSO wells - ave blank wells) × 100%.

[0142] Wherein, ave compound hole represents the average emission signal intensity of the compound hole; ave blank hole represents the average emission signal intensity of the blank hole; and ave DMSO hole represents the average emission signal intensity of the DMSO hole.

[0143] To accurately compare the activities of each compound, the concentration of the compound in the dilution solution was selected to be 500 nM. The inhibition rate results at this concentration are shown in Table 2.

[0144] Data Analysis: Using the concentration log₂ as the X-axis and the percentage inhibition rate as the Y-axis, the dose-response curve was fitted using the "[Inhibitor] vs. normalized response -- Variable slope" model in GraphPad Prism 9 software to obtain the IC₂ of each compound on enzyme activity. 50 value. Table 2 shows the inhibition rates of the compounds on the in vitro NUAK1 and NUAK2 kinase activities.

[0145] As shown in Table 2, the compounds of this invention can effectively inhibit the activities of NUAK1 and NUAK2 enzymes. Furthermore, the IC50 values ​​of some compounds of this invention for inhibiting the activities of NUAK1 and NUAK2 enzymes in vitro were determined. 50 The values ​​are shown in Table 3.

[0146] Table 3. IC50 values ​​of the compounds for in vitro inhibition of NUAK1 and NUAK2 enzyme activities. 50 value

[0147] As shown in Table 3, the compounds of the present invention have good inhibitory activity against NUAK1 and NUAK2 kinases.

[0148] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the patent scope of the present invention. All equivalent implementations or modifications that do not depart from the present invention should be included within the scope of the technical solution of the present invention.

Claims

1. An aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs, characterized in that, The structural formula of the aminopyrimidine compound is shown in formula (I): Formula (I) in, Selected from , , , , , , , , , or ; Selected from or ; The wavy line indicates the connection point.

2. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 1, characterized in that, R1 is selected from -H, -C 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2; and / or n is an integer selected from 1 or 2; and / or X is selected from -NH-, -O-, or a chemical bond.

3. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 1, characterized in that, R a Selected from -H, -C 1-3 Alkyl, -OC 1-3 Alkyl or -OCHF2; And / or when R c When it is H, R b Selected from C 4-8 Heterocyclic alkanes or C 4-10 Bridged cyclic alkane group, the C 4-8 Heterocyclic alkanes or C 4-10 The bridged cycloalkane group optionally contains one, two, or three additional heteromers selected from N, O, and S, and is optionally separated by a group selected from H, halogen, carbonyl, -C. 1-3 Alkyl groups, -N(CH3)2 substituents; when R c When R is not H, b and R c It forms a ring C with the atoms in between; and / or R d Selected from ring C; And / or U and V are each independently selected from CH or N; Wherein, the ring C is selected from C 4-6 cycloalkyl or C 4-6 Contains an N-heterocyclic alkyl group; and the ring C is optionally separated by one or more elements selected from -H, halogen, -C. 1-3 Substitution of alkyl, -OH, and carbonyl groups.

4. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 2, characterized in that, Selected from , , , , , , , , , , or .

5. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 3, characterized in that, R a Selected from -H, -OC 1-3 Alkyl or -OCHF2; And / or when R c When it is H, R b Selected from , , , , , , , or When R c When it is not H, Selected from or ; and / or R d Selected from or .

6. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 5, characterized in that, The structural formula of the aminopyrimidine compound is shown in formula (II): Equation (II) in, Selected from , , , , , , , , , , or ; R1 is selected from -H, -C 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2; n is an integer selected from 1 or 2; X is selected from -NH-, -O-, or chemical bonds; R b Selected from , , or .

7. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 6, characterized in that, Selected from , , , , , , , , , , or .

8. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 5, characterized in that, The structural formula of the aminopyrimidine compound is shown in formula (III): Equation (III) in, Selected from , or R1 is selected from -H, -C 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2; n is an integer selected from 1 or 2; X is selected from -NH- or chemical bonds.

9. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 1, characterized in that, The structural formula of the aminopyrimidine compound is shown in formula (IV): Formula (IV) in, Selected from or ; R1 is selected from -H, -C 1-3 Alkyl, -CN, -CONH2, carbonyl, or -NH2; W is selected from C or N; R2 is selected from H or -C 1-3 alkyl.

10. The aminopyrimidine compound or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs according to claim 1, characterized in that, The aminopyrimidine compound is selected from any one of the following compounds: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 11. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises an aminopyrimidine compound as described in any one of claims 1-10, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, prodrug, and a pharmaceutically acceptable carrier thereof.

12. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises an aminopyrimidine compound as described in any one of claims 1-10, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, prodrug, and other pharmaceuticals.

13. The pharmaceutical composition according to claim 12, characterized in that, The other drugs are selected from one or more of paclitaxel, carboplatin, cisplatin, capecitabine, irinotecan, and azacitidine.

14. The use of the aminopyrimidine compound of any one of claims 1-10, or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, or the pharmaceutical composition of any one of claims 11-13, in the preparation of a medicament for treating diseases caused by abnormal activation of NUAK kinase.

15. The application according to claim 14, characterized in that, The NUAK kinases include NUAK1 kinase and / or NUAK2 kinase.

16. The application according to claim 14, characterized in that, The disease is selected from one or more of the following: autoimmune diseases, inflammatory diseases, visceral fibrosis, skin fibrosis, and cancer.

17. The application according to claim 16, characterized in that, The cancer is selected from one or more of the following: gastrointestinal stromal tumors, esophageal cancer, gastric cancer, melanoma, glioma, glioblastoma, ovarian cancer, bladder cancer, pancreatic cancer, prostate cancer, lung cancer, breast cancer, kidney cancer, liver cancer, osteosarcoma, multiple myeloma, cervical cancer, bone metastases, papillary thyroid carcinoma, non-small cell lung cancer, and colorectal cancer.

18. The application according to claim 16, characterized in that, The disease is selected from one or more of the following: rheumatoid arthritis, inflammatory bowel disease, chronic obstructive pulmonary disease, asthma, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, liver fibrosis, renal fibrosis, sequelae of myocarditis, scleroderma, keloids, hypertrophic scars, traumatic and postoperative scars.

19. The use of the aminopyrimidine compound of any one of claims 1-10, or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, prodrugs, or the pharmaceutical composition of any one of claims 11-13, in the preparation of a NUAK kinase inhibitor.

20. An intermediate compound, characterized in that, Including any one of the following compounds: 、 、 、 、 、 、 、 、 、 、 、 、 、 。

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