CN107619407A - Double target spot inhibitor of HDAC and VEGFR based on pazopanib structure and its preparation method and application - Google Patents

Abstract

The invention relates to a dual-target inhibitor of HDAC and VEGFR based on the structure of pazopanib, a preparation method and application thereof. The compound has a structure represented by general formula I, II or III. The invention also provides a preparation method of the compound and its application in the preparation of drugs for preventing or treating tumor-related diseases.

Description

HDAC and VEGFR dual-target inhibitor based on pazopanib structure and preparation method thereof law and application

technical field

The invention belongs to the technical field of organic compound synthesis and medical application, and specifically relates to a dual-target inhibitor of HDAC and VEGFR based on the structure of pazopanib and its preparation method and application.

Background technique

Pazopanib is a multi-target receptor tyrosine kinase inhibitor for the three subtypes of vascular endothelial growth factor receptor VEGFR (VEGFR-1, VEGFR-2, VEGFR-3) and associated receptors All body tyrosine kinases (PDGFRβ, c-Kit, FGF-R1, c-fms) showed good inhibitory activity (IC ₅₀ : 10, 30, 47, 84, 74, 140, 146 nM) (Harris, pHilip A.etc ., Journal of Medicinal Chemistry 2008, 51, 4632). Pazopanib was approved by the US Food and Drug Administration FDA in October 2009 for the treatment of advanced renal cell carcinoma in patients with previous chemotherapy experience (Bukowski.etc., Nature Reviews Drug Discovery, 2010,9,17) , was approved by the FDA in April 2012 for the treatment of advanced soft tissue sarcoma (Wilky, Breelyn A.etc., Currentopinion in oncology, 2013, 25, 373). However, in recent years, the low response rate and drug resistance of pazopanib have appeared in the clinical application process (Gotink K J.etc., Cellular Oncology, 2015, 38, 119), drug combination application and development of multi-target drugs are A strategy to enhance tumor sensitivity to drugs and reduce tumor resistance.

Histone deacetylases (HDACs) are a group of enzymes that regulate the level of histone acetylation in vivo. The level of histone acetylation has an important impact on the structure of chromatin and the transcriptional regulation of genes. The mutation and abnormal expression of HDACs are usually associated with the development of tumors. occur closely related. HDACs inhibitors have been proven to have anti-tumor effects, and currently 5 small molecule HDAC inhibitors have been approved for marketing for the treatment of various hematological tumors (Li X.etc., Current Drug Targets, 2014, 15, 622). With the gradual elucidation of the relationship between HDAC and VEGF/VEGFR signaling pathway, the combined application of pazopanib and HDAC inhibitors has aroused the interest of researchers. The study found that the combination of pazopanib and HDAC inhibitors (VPA, SAHA) exhibited additive or synergistic anti-tumor effects in vivo and in vitro in soft tissue sarcoma, and the combined drug could reverse pazopanib-resistant tumor lines Drug resistance (Tavallai S.etc., Cancer Biology & Therapy, 2014, 15, 578). The combination of HDAC inhibitor AR-42 and pazopanib showed killing effect on dabrafenib/trametinib-resistant melanoma in vivo and in vitro experiments, and its synergistic effect and activation of multiple signals pathway (Booth L.etc., Oncotarget, 2017, 8, 16367). A clinical phase I study reported in 2015 showed that pazopanib combined with HDAC inhibitor SAHA showed significant curative effect on tumor patients with TP53 mutation, especially metastatic sarcoma and metastatic colorectal cancer. Progression-free survival and median overall survival were significantly prolonged (Fu S.etc., Annals of Oncology, 2015, 26, 1012). A clinical phase I study newly reported in 2017 showed that the HDAC inhibitor abexinost combined with pazopanib has good tolerance and can effectively overcome pazopanib resistance (Aggarwal, Rahul, etc., Journal of Clinical Oncology, 2017, 35, 1231).

Clinically, the combined application of antineoplastic drugs with different mechanisms of action has been considered as a standard solution to avoid drug tolerance, but the following problems exist in the combined drug: (1) The pharmacokinetics of the combined drug is complex, drug-drug interactions, adverse The response is difficult to predict; (2) The compatibility and dosage setting of different drugs are clinically difficult. Multi-target drug refers to a single drug molecule that acts on multiple targets at the same time. Multi-target drug not only has the advantages of combination drug, but also overcomes some defects of combination drug. Multi-target drugs have simple pharmacokinetics, avoid drug-drug interactions, are safer, and improve patient compliance, which has become a popular direction in the design of anticancer drugs. At present, there is no relevant report in the prior art on HDAC and VEGFR dual-target inhibitors based on the structure of pazopanib.

Contents of the invention

Aiming at the deficiencies of the prior art, the present invention provides a dual-target inhibitor of HDAC and VEGFR based on the structure of pazopanib, and the present invention also provides a preparation method and use of the above compound.

Technical scheme of the present invention is:

1. HDAC and VEGFR dual-target inhibitor based on pazopanib structure

The present invention contains the HDAC/VEGFR dual-target inhibitor based on the pazopanib structure, and its pharmaceutically acceptable salt, solvate or prodrug has the structure shown in the following general formula I, II or III:

in:

X is

n is 0-9;

Y is

Preferably according to the invention,

Y in general formula I and X in II are in para-position or meta-position;

X is

n is 0-7;

Y is

Further preferably, the above compound is one of the following:

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-hydroxybenzamide (6a),

3-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-hydroxybenzamide (6b),

5-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-hydroxy-2-methylbenzyl Amide (6c),

N-(2-aminophenyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene Formamide (6d),

N-(2-aminophenyl)-3-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene Formamide (6e),

(E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)- N-hydroxyacrylamide (10a),

(E)-3-(3-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)- N-hydroxyacrylamide (10b),

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(2-(hydroxylamine)-2 -Oxoethyl)benzamide (13a),

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(3-(hydroxylamine)-3 -Oxopropyl)benzamide (13b),

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(4-(hydroxylamine)-4 -Oxobutyl)benzamide (13c),

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(5-(hydroxylamine)-5 -oxopentyl)benzamide (13d),

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(6-(hydroxylamine)-6 -oxohexyl)benzamide (13e),

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(7-(hydroxylamine)-7 -Oxoheptyl)benzamide (13f),

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(8-(hydroxylamine)-8 -Oxooctyl)benzamide (13g),

3-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(6-(hydroxylamine)-6 -oxohexyl)benzamide (13h),

5-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(6-(hydroxylamine)-6 -oxohexyl)-2-methylbenzamide (13i),

N-(2-((2-aminophenyl)amino)-2-oxoethyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( Methyl)amino)pyrimidin-2-yl)amino)benzamide (14a),

N-(3-((2-aminophenyl)amino)-3-oxopropyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( Methyl)amino)pyrimidin-2-yl)amino)benzamide (14b),

N-(4-((2-aminophenyl)amino)-4-oxobutyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( Methyl) amino) pyrimidin-2-yl) amino) benzamide (14c),

N-(5-((2-aminophenyl)amino)-5-oxopentyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( Methyl) amino) pyrimidin-2-yl) amino) benzamide (14d),

N-(6-((2-aminophenyl)amino)-6-oxohexyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)(form Base) amino) pyrimidin-2-yl) amino) benzamide (14e),

N-(7-((2-aminophenyl)amino)-7-oxoheptyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( Methyl) amino) pyrimidin-2-yl) amino) benzamide (14f),

N-(8-((2-aminophenyl)amino)-8-oxooctyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( Methyl)amino)pyrimidin-2-yl)amino)benzamide (14g),

2-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy Acetamide (19a),

3-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy Propionamide (19b),

4-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy Butanamide (19c),

5-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy Valeramide (19d),

6-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy Caproamide (19e),

7-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy Heptanamide (19f),

8-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy Caprylamide (19g),

N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ³ - Hydroxymalonamide (23a),

N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ⁴ - Hydroxysuccinamide (23b),

N ¹ -(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ⁵ - Hydroxyglutaramide (23c),

N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ⁶ - Hydroxyadipamide (23d),

N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ⁷ - Hydroxypimelicamide (23e),

N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ⁸ - Hydroxysuberamide (23f),

N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ⁹ - Hydroxyazalamide (23g),

2-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -hydroxyacetamide (28a)

3-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -Hydroxypropionamide (28b)

4-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -Hydroxybutanamide (28c)

5-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -Hydroxyvaleramide (28d)

6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -Hydroxycaproamide (28e)

7-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -Hydroxyheptanamide (28f) or

8-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -Hydroxyoctanoamide (28 g).

2. Preparation method of HDAC and VEGFR dual-target inhibitor based on pazopanib structure

The preparation method of the present invention containing the HDAC and VEGFR dual-target inhibitor based on the pazopanib structure is one of the following methods:

(1) Using 2,3-dimethyl-6-amino-2H-indazole as the starting material, it undergoes a nucleophilic substitution reaction with 2,4-dichloropyrimidine under sodium bicarbonate alkaline conditions to obtain intermediate 2 , intermediate 2 reacts with methyl iodide under the catalysis of cesium carbonate to obtain intermediate 3, and intermediate 3 undergoes nucleophilic substitution reaction with different substituted anilines under the catalysis of concentrated hydrochloric acid to obtain key intermediate 4, which is protected by methyl ester The intermediate 5 was obtained, and the final product 6a-6c was obtained through the ammonolysis reaction of the intermediate 5; in addition, the intermediate 4a-4b was condensed with o-phenylenediamine to obtain the final product 6d-6e.

The reaction formula is as follows:

Among them, the benzene rings of 4a, 5a, 6a, and 6d are para-substituted, and the benzene rings of 4b, 5b, 6b, and 6e are meta-substituted.

Reagents and conditions in the above reaction formula: (a) 2,4-dichloropyrimidine, sodium bicarbonate, absolute ethanol, reflux, 4h; (b) cesium carbonate, methyl iodide, N,N-dimethylformamide , room temperature, 2h; (c) isopropanol, concentrated hydrochloric acid, reflux, 4h; (d) O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, triethyl Amine, o-phenylenediamine, N,N-dimethylformamide, 0°C-room temperature, overnight; (e) anhydrous methanol, thionyl chloride, reflux, overnight; (f) hydroxylamine hydrochloride, potassium hydroxide, Anhydrous methanol, room temperature, 2h.

The different substituted anilines are: p-aminobenzoic acid, m-aminobenzoic acid or 5-amino-2-methylbenzoic acid.

Preferably according to the present invention, the specific preparation steps are as follows:

4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-hydroxybenzamide (6a) Preparation method, the steps are as follows:

(1) Preparation of N-(2-chloropyrimidin-4-yl)-2,3-dimethyl-6-amino-2H-indazole (2)

Dissolve 5.00g of 2,3-dimethyl-6-amino-2H-indazole, 13.9g of 2,4-dichloropyrimidine, and 10.4g of sodium bicarbonate in 100mL of absolute ethanol, and heat to 79°C. After refluxing for 4 hours, cool down to room temperature, filter, wash the filter cake fully with ethyl acetate, collect the filtrate, concentrate under reduced pressure, remove the solvent, fully beat with ethyl acetate, filter to obtain an off-white solid, and recrystallize with methanol to obtain 7.64g Product 2, as a white solid.

(2) Preparation of N-(2-chloropyrimidin-4-yl)-N,2,3-trimethyl-2H-indazol-6-amine (3)

Add 1.00g of product 2 from the previous step and 2.40g of cesium carbonate to 50mL N,N-dimethylformamide, react at room temperature for 20min, then slowly add 0.78g of methyl iodide, after the addition is complete, react at room temperature for 2h, pour the reaction solution into A large amount of light yellow solid precipitated immediately in ice water, filtered, dried, and recrystallized with ethyl acetate to obtain 1.06 g of product 3 as light yellow crystals.

(3) Preparation of 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoic acid (4a)

Disperse 0.50g of intermediate 3 and 0.36g of p-aminobenzoic acid in 30mL of isopropanol, add 2 drops of concentrated hydrochloric acid, heat to 85°C, reflux for 4h, cool to room temperature, filter, and wash with isopropanol and ethyl acetate The filter cake, dried yielded 0.53 g of product 4a as a white solid.

(4) Methyl 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoate (5a ) preparation

Add 0.50g of 4a into 50mL of anhydrous methanol, then add 0.60g of thionyl chloride dropwise at 0°C, after the addition is complete, keep warm for 30min, remove the ice bath and heat to reflux, react for 5h, and depressurize the solvent After evaporation, the crude product was recrystallized from isopropanol to give 0.50 g of product 5a as a white solid.

(5) 4-((4-((2,3-dimethyl-2H-indazol-6-yl) (methyl) amino) pyrimidin-2-yl) amino)-N-hydroxybenzamide ( 6a) Preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.20 g of compound 5a in 30 mL potassium hydroxylamine solution, stir at room temperature for 1 h, concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 80.6 mg Product 6a, as a white solid.

N-(2-aminophenyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene The preparation method of formamide (6d), the steps are as follows:

(1) N-(2-aminophenyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl) Preparation of amino) benzamide (6d)

Add 0.50 g of intermediate 4a to 50 mL of anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethylurea Fluoroboric acid 0.49g and triethylamine 0.26g, activated for 30 minutes, then add o-phenylenediamine 0.17g, react overnight at room temperature, add ethyl acetate to dilute after the reaction is completed, wash to remove N,N-dimethylformamide , the ethyl acetate layer was washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography to obtain 0.40 g of product 6d as an off-white solid.

(2) Using intermediate 4 as the starting material, it undergoes amide condensation with N,O-dimethyl hydroxylamine hydrochloride to obtain intermediate 7, which is reduced to aldehyde intermediate 8 by tetrahydrolithium aluminum, and intermediate 8 is combined with phosphorus Triethyl acyl acetate undergoes Horner-Wadsworth-Emmons reaction to obtain intermediate 9, and intermediate 9 undergoes ester aminolysis to obtain final product 10.

The reaction formula is as follows:

Among them, the benzene rings of 4a, 7a, 8a, 9a, and 10a are para-substituted, and the benzene rings of 4b, 7b, 8b, 9b, and 10b are meta-substituted.

Reagents and conditions in the above reaction formula: (a) N,O-dimethylhydroxylamine hydrochloride, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethyl Amine, N,N-dimethylformamide, room temperature, overnight; (b) lithium aluminum tetrahydrogen, tetrahydrofuran, -20°C, 4h; (c) triethyl phosphoroacetate, sodium hydride, tetrahydrofuran, 0°C- Room temperature, overnight; (d) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h.

Preferably according to the present invention, the specific preparation steps are as follows:

(E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)- Preparation of N-hydroxyacrylamide (10a):

(1) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-methoxy-N -Preparation of methylbenzamide (7a)

Add 1.00 g of intermediate 4a to anhydrous N,N-dimethylformamide, under ice-cooling, add O-benzotriazole-N,N,N',N'-tetramethylurea Fluoroboric acid 0.98g and triethylamine 0.52g, activated for 30min, then add N,O-dimethyl hydroxylamine hydrochloride 0.30g and triethylamine 0.31g, react overnight at room temperature, add ethyl acetate to dilute after the reaction is completed, wash with water Remove N,N-dimethylformamide, wash the ethyl acetate layer with saturated sodium bicarbonate solution and saturated NaCl solution, dry over anhydrous magnesium sulfate, remove the solvent under reduced pressure, and use dichloromethane:methanol=35:1 for the residue The volume ratio of the eluent was purified by column chromatography to obtain 0.78 g of product 7a as a white solid.

(2) Preparation of 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzaldehyde (8a)

1.00 g of compound 7a was dissolved in 50 mL of anhydrous tetrahydrofuran, and cooled to -40 °C in a cryogenic reactor. Slowly add 0.26 g of lithium aluminum tetrahydrogen to it in batches, and maintain the temperature for 2 hours. After the reaction is completed, slowly and carefully quench the reaction with ice water until no more bubbles are generated. The reaction solution was diluted with water, extracted with ethyl acetate, the organic layers were combined, washed with saturated NaCl solution, dried over anhydrous magnesium sulfate, the solvent was concentrated under reduced pressure, and the residue was subjected to column separation with dichloromethane:methanol=40:1 volume ratio eluent Purification by chromatography afforded 0.61 g of product 8a as a white solid.

(3) Ethyl (E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino ) Preparation of phenyl) acrylate (9a)

Add 0.32 g of triethyl phosphoroacetate to 50 mL of anhydrous tetrahydrofuran, cool to -5-0 °C, slowly add 0.16 g of NaH, after the addition is complete, stir for 20 min, then add 0.52 g of 8a in tetrahydrofuran dropwise, and react overnight at room temperature. After the reaction was completed, 50 mL of 10% ammonium chloride solution was added, stirred for 30 min, and then separated into layers. After the organic phase was dried, filtered, and concentrated to obtain a crude product. The residue was purified by column chromatography with dichloromethane:methanol=40:1 volume ratio eluent to obtain 0.48 g of product 9a as a white solid.

(4) (E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene base)-N-hydroxyacrylamide (10a) preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Compound 9a 0.50g, dissolved in 30mL potassium hydroxylamine solution, stirred at room temperature for 2h, concentrated under reduced pressure to remove most of the solvent, the remaining residue was adjusted to pH 5-6 with 1M HCl, a white solid was precipitated, filtered, and the filter cake was dried to obtain 0.29 g Product 10a, as a white solid.

(3) Using intermediate 4 as the starting material, intermediate 4 and aminoalkane methyl esters of different chain lengths undergo amide condensation reaction to obtain intermediate 11, and intermediate 11 obtains final product 13 through the aminolysis reaction of ester; intermediate 11 was hydrolyzed to intermediate 12 under alkaline conditions of sodium hydroxide, and intermediate 12 was condensed with o-phenylenediamine to obtain final product 14 through amide condensation.

The reaction formula is as follows:

Wherein 4a, 11a-11g, 12a-12g, 13a-13g, 14a-14g have para-substituted benzene rings, 4b, 11h, 13h have meta-substituted benzene rings, and n is 1-7.

Reagents and conditions in the above reaction formula: (a) aminoalkane methyl esters with different chain lengths, O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, triethyl Amine, N,N-dimethylformamide, room temperature, overnight; (b) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h; (c) 3M NaOH, methanol, room temperature, 2h; (d) o Phenylenediamine, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, N,N-dimethylformamide, room temperature, overnight.

The aminoalkane methyl esters of different chain lengths are: glycine methyl ester, 3-aminopropionic acid methyl ester, 4-aminobutyric acid methyl ester, 5-aminovaleric acid methyl ester, 6-aminocaproic acid methyl ester, Methyl 7-aminoheptanoate or methyl 8-aminooctanoate.

Preferably according to the present invention, the specific preparation steps are as follows:

4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(2-(hydroxylamine)-2 -The preparation method of oxoethyl)benzamide (13a), the steps are as follows:

(1) Methyl (4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoyl)glycinate (11a) preparation of

Add 1.00 g of intermediate 4a to anhydrous N,N-dimethylformamide, under ice-cooling, add O-benzotriazole-N,N,N',N'-tetramethylurea Fluoroboric acid 0.98g and triethylamine 0.52g, activated for 30 minutes, then added glycine methyl ester hydrochloride 0.39g, reacted overnight at room temperature, after the reaction was completed, added ethyl acetate to dilute, washed with water to remove N,N-dimethyl The formamide and ethyl acetate layers were washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure, and the residue was subjected to column chromatography with dichloromethane:methanol=30:1 volume ratio as the eluent Analytical purification afforded 0.87 g of product 11a as a white solid.

(2) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(2-(hydroxylamine )-2-oxoethyl) benzamide (13a) preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Compound 11a 0.50g was dissolved in 30mL potassium hydroxylamine solution, stirred at room temperature for 2h, concentrated under reduced pressure to remove most of the solvent, and the remaining residue was adjusted to pH 5-6 with 1M HCl, a white solid was precipitated, filtered, and the filter cake was dried to obtain 0.31g Product 13a, as a white solid.

N-(2-((2-aminophenyl)amino)-2-oxoethyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( The preparation of methyl) amino) pyrimidin-2-yl) amino) benzamide (14a), the steps are as follows:

(1) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoyl)glycine (12a) preparation of

Add 1.00 g of 11a to 30 mL of methanol, then add 5 mL of 3M NaOH solution, and stir at room temperature for 4 h. After the reaction, most of the methanol is evaporated under reduced pressure, and the residue is adjusted to pH 5-6 with 1 M HCl solution. After filtration, the filter cake was washed with ice water, and after drying, 0.81 g of product 12a was obtained as a white solid, which was directly used for the next step.

(2) N-(2-((2-aminophenyl)amino)-2-oxoethyl)-4-((4-((2,3-dimethyl-2H-indazole-6- base) (methyl) amino) pyrimidin-2-yl) amino) benzamide (14a) preparation

Add 1 g of intermediate 12a to 100 mL of anhydrous N,N-dimethylformamide, under ice-cooling, add O-benzotriazole-N,N,N',N'-tetramethylurea tetra Fluoroboric acid 0.86g and triethylamine 0.45g, activated for 30min, then add o-phenylenediamine 0.29g, react overnight at room temperature, add ethyl acetate to dilute after the reaction is completed, wash to remove N,N-dimethylformamide , the ethyl acetate layer was washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=30:1 volume ratio eluent 0.47 g of product 14a was obtained as a pale yellow solid.

(4) Using p-nitrophenol as the starting material, nucleophilic substitution reaction occurs with methyl bromoalkanoate of different chain lengths to obtain intermediate 16, intermediate 16 is reduced by Pd/C to obtain intermediate 17, intermediate 17 and Compound 3 undergoes a nucleophilic reaction to obtain intermediate 18, and intermediate 18 undergoes ester ammonolysis to obtain final product 19.

The reaction formula is as follows:

wherein n is 1-7.

Reagents and conditions in the above reaction: (a) methyl bromoalkanoate with different chain lengths, potassium carbonate, acetone, reflux, overnight; (b) Pd/C, methanol, hydrogen, room temperature, overnight; (c) isopropanol , concentrated hydrochloric acid, reflux, overnight; (d) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h.

The methyl bromoalkanoates of different chain lengths are: methyl 2-bromoacetate, methyl 3-bromopropionate, methyl 4-bromobutyrate, methyl 5-bromovalerate, 6-bromohexanoic acid methyl ester, methyl 7-bromoheptanoate, or methyl 8-bromooctanoate.

Preferably according to the present invention, the specific preparation steps are as follows:

7-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)-N-hydroxy The preparation of heptanamide (19f), the steps are as follows:

(1) Preparation of methyl 7-(4-nitrophenoxy)heptanoate (16f)

Add 1.00g of p-nitrophenol, 1.93g of methyl 7-bromoheptanoate and 2.98g of potassium carbonate to 50mL of acetone, raise the temperature to reflux, and react overnight. Crystallization afforded 1.62 g of product 16f as a yellow solid.

(2) Preparation of methyl 7-(4-aminophenoxy)heptanoate (17f)

Add 1.00 g of intermediate 16f to 50 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, react overnight at room temperature, filter, wash the filter residue with methanol, combine the filtrate and washing liquid, and evaporate under reduced pressure Removal of the solvent afforded 0.79 g of product 17f as a light brown solid.

(3) Methyl 7-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy )-Heptanoic acid ester (18f) preparation

Add 0.5 g of intermediate 3, 0.52 g of intermediate 17f and 0.5 mL of concentrated hydrochloric acid into 50 mL of isopropanol at room temperature, raise the temperature to reflux, and react overnight. After the reaction, the solvent was evaporated under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=40:1 volume ratio eluent to obtain 0.61 g of product 18f as a white solid.

(4) 7-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)- Preparation of N-Hydroxyheptanamide (19f)

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 h, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.30 g of compound 18f in 30 mL of potassium hydroxylamine solution, stir at room temperature for 2 h, concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 0.20 g Product 19f, as a white solid.

(5) Using intermediate 3 as the starting material, nucleophilic substitution reaction occurs with p-nitroaniline to obtain intermediate 20. Intermediate 20 is reduced to intermediate 21 by Pd/C. Acid monomethyl ester undergoes amide condensation to obtain intermediate 22, and intermediate 22 undergoes ester ammonolysis to obtain final product 23.

The reaction formula is as follows:

wherein n is 1-7.

Reagents and conditions for the above reaction: (a) p-nitroaniline, isopropanol, concentrated hydrochloric acid, reflux, overnight; (b) Pd/C, methanol, hydrogen, room temperature, overnight; (c) alkanediol with different chain lengths Acid monomethyl ester, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, N'N-dimethylformamide, room temperature, overnight; ( d) Hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h.

The monomethyl alkanedioic acid of described different chain lengths is: monomethyl malonate, monomethyl succinate, monomethyl glutarate, monomethyl adipate, monomethyl pimelate, Monomethyl Suberate or Monomethyl Azelate.

Preferably according to the present invention, the specific preparation steps are as follows:

N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)-N ⁸ - The preparation of hydroxysuberamide (23f), the steps are as follows:

(1) N ⁴ -(2,3-dimethyl-2H-indazol-6-yl)-N ⁴ -methyl-N ² -(4-nitrophenyl)pyrimidine-2,4-diamine Preparation of (20)

Add 1.00g of intermediate 3, 0.58g of p-nitroaniline and 0.5mL of concentrated hydrochloric acid to 70mL of isopropanol, raise the temperature to reflux, and react overnight. After the reaction, cool to room temperature, filter, and filter the cake with a small amount of isopropanol Washing and drying of the filter cake yielded 1.08 g of product 20 as a pale yellow solid.

(2) N ² -(4-aminophenyl)-N ⁴ -(2,3-dimethyl-2H-indazol-6-yl)-N ⁴ -methylpyrimidine-2,4-diamine ( 21) Preparation

Suspend 1.00 g of intermediate 20 in 70 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, and react overnight at room temperature. After the reaction, the solvent is evaporated under reduced pressure, and the residue is recrystallized with ethyl acetate 0.78 g of product 21 was obtained as a light brown solid.

(3) Methyl 8-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl )amino)-8-oxooctyl ester (22f) preparation

Add 0.33g of monomethyl suberate to 50mL of anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethyl After adding 0.68g of urea tetrafluoroboric acid and 0.36g of triethylamine, activate in ice bath for 30min. After activation, add 0.76g of intermediate 21, remove the ice bath, and react overnight at room temperature. After the reaction is completed, add ethyl acetate to dilute. Wash with water to remove N,N-dimethylformamide, wash the ethyl acetate layer with saturated sodium bicarbonate solution and saturated NaCl solution, dry over anhydrous magnesium sulfate, distill off the solvent under reduced pressure, and wash the residue with dichloromethane:methanol=30 Purification by column chromatography with an eluent in a volume ratio of :1 gave 0.37 g of product 22 as a white solid.

(4) N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)- Preparation of N ⁸ -Hydroxysuberamide (23f)

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.20 g of compound 22f in 30 mL potassium hydroxylamine solution, stir at room temperature for 2 h, concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 0.14 g Product 23f, as a white solid.

(6) Using p-fluoronitrobenzene as the starting material, nucleophilic substitution reaction with aminoalkanoic acid methyl esters of different chain lengths produces intermediate 25, intermediate 25 is reduced by Pd/C to obtain intermediate 26, intermediate 26 A nucleophilic substitution reaction with intermediate 3 gave intermediate 27, and intermediate 27 was subjected to ester ammonolysis to give intermediate 28.

The reaction formula is as follows:

wherein n is 1-7.

Reagents and conditions for the reaction: (a) methyl aminoalkanoate with different chain lengths, potassium carbonate, N,N-dimethylformamide, 50°C, overnight; (b) Pd/C, hydrogen, methanol, overnight; (c) concentrated hydrochloric acid, isopropanol, reflux, overnight; (d) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h.

The aminoalkanoic acid methyl esters of different chain lengths are: glycine methyl ester, 3-aminopropionic acid methyl ester, 4-aminobutyric acid methyl ester, 5-aminovaleric acid methyl ester, 6-aminocaproic acid methyl ester, Methyl 7-aminoheptanoate or methyl 8-aminooctanoate.

Preferably according to the present invention, the specific preparation steps are as follows:

6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino)-N -The preparation of hydroxy caproamide (28e), the steps are as follows:

(1) Preparation of methyl 6-((4-nitrophenyl)amino)hexyl ester (25e)

Add 1.00g of p-fluoronitrobenzene, 1.54g of methyl 6-aminocaproate hydrochloride and 2.94g of potassium carbonate into 50mL of N,N-dimethylformamide, raise the temperature to 50°C overnight, and after the reaction , cooled to room temperature, the reaction solution was poured into ice water, a yellow solid was precipitated, filtered, dried, and the residue was subjected to column chromatography with petroleum ether:ethyl acetate=4:1 volume ratio eluent to obtain 1.32g of product 25e, It is light yellow solid.

(2) Preparation of methyl 6-((4-aminophenyl)amino)hexyl ester (26e)

Suspend 1.00 g of intermediate 25e in 70 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, and react overnight at room temperature. After the reaction, evaporate the solvent under reduced pressure, and weigh the residue with ethyl acetate. Crystallization afforded 0.71 g of product 26e as a brown solid.

(3) Methyl 6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl )amino)-hexyl ester (27e) preparation

Add 0.50 g of intermediate 3, 0.49 g of intermediate 26e and 0.5 mL of concentrated hydrochloric acid into 50 mL of isopropanol at room temperature, raise the temperature to reflux, and react overnight. After the reaction, the solvent was evaporated under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=40:1 volume ratio eluent to obtain 0.55 g of product 27e as a light yellow solid.

(4) 6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino )-N-hydroxy caproamide (28e) preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.20 g of compound 27e in 30 mL of potassium hydroxylamine solution, stir at room temperature for 2 h, then concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 0.13 g Product 28e, as a reddish solid.

The structural formula of the target compound is as follows:

The specific preparation steps of the compound will be described in detail in the examples.

Those skilled in the art can optimize the above steps to improve the yield. They can formulate synthetic routes based on the basic knowledge in the art, such as selecting reactants, solvents and temperatures, and can avoid the occurrence of side reactions by using various protecting groups. Thereby increasing the yield. These conventional protection methods can be found in T. Greene, Protecting Groups in Organic Synthesis.

Detailed description of the invention

The terms and definitions used in this document have the following meanings:

ACHN cells are human renal cell adenocarcinoma cells, AGS cells are human gastric adenocarcinoma cells, HEL cells are human erythroleukemia cells, HeLa cells are human cervical cancer cells, HT-1080 cells are human fibrosarcoma cells, HT-29 cells are human colon cancer cells, K562 The cells are human chronic myeloid leukemia cells, KG1 cells are human acute myeloid leukemia cells, MDA-MB-231 are human breast cancer cells, MOLT-4 are human acute lymphoblastic leukemia cells, PC-3 cells are human prostate cancer cells and HUVEC cells Umbilical vein endothelial cells.

3. Application of HDAC and VEGFR dual-target inhibitors based on pazopanib structure

The present invention also provides the application of the HDAC and VEGFR dual-target inhibitor compound based on the structure of pazopanib in the preparation of drugs for preventing or treating tumor-related diseases.

The tumor-related diseases include various hematological tumors and various solid tumors.

The various hematological tumors include: various leukemias and myeloproliferative tumors. Various types of leukemia such as severe acute lymphoblastic leukemia, acute myeloid leukemia, acute megakaryoblastic leukemia, etc. Myeloproliferative neoplasms include chronic myelogenous leukemia, polycythemia vera, essential thrombocythemia, myelofibrosis, chronic neutrophil leukemia, chronic eosinophilic leukemia, systemic mastocytosis, and other bone marrow disorders not otherwise classified Hyperplasia.

The solid tumors include: various forms of nasopharyngeal carcinoma, renal cell carcinoma, soft tissue sarcoma, papillary thyroid carcinoma, thymoma, liver cancer, breast cancer, melanoma, prostate cancer, retinoblastoma, and the like.

A pharmaceutical composition for preventing or treating tumor-related diseases, comprising the HDAC and VEGFR dual-target inhibitor based on the pazopanib structure of the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically Carriers or excipients are acceptable.

Description of drawings

Figure 1 Western Blot experiment results of compounds 6d and 13f after HeLa cells were treated for 4 hours;

Figure 2 Compound 6d and 13f inhibit HUVECs lumen formation experimental results;

Fig. 3 compound 6d and 13f inhibit rat thoracic aorta angiogenesis experiment result;

Fig. 4 photo of tumor in nude mice;

Fig. 5 Inhibitory rate of tumor growth of compounds 6d and 13f at different doses.

Detailed ways

The present invention will be further described below in conjunction with examples, but not limited thereto.

Example 1: Synthesis of compounds 6a-6c, taking 6a as an example.

(1) Preparation of N-(2-chloropyrimidin-4-yl)-2,3-dimethyl-6-amino-2H-indazole (2)

Dissolve 5.00g of 2,3-dimethyl-6-amino-2H-indazole, 13.9g of 2,4-dichloropyrimidine and 10.4g of sodium bicarbonate in 100mL of absolute ethanol, heat to 79°C, and reflux for 4h After cooling down to room temperature, filter, the filter cake was fully washed with ethyl acetate, the filtrate was collected, concentrated under reduced pressure, the solvent was removed, fully beaten with ethyl acetate and filtered to obtain an off-white solid, which was recrystallized with methanol to obtain 7.64g of product 2 , as a white solid, yield: 90.0%; Mp: 215-216°C. ¹ H NMR (300MHz, DMSO-d ₆ ) δ9.99(s, 1H), 8.15(d, J=5.9Hz, 1H), 7.94(s,1H),7.63(dd,J=0.8,8.9Hz,1H),6.98(dd,J=1.8,8.9Hz,1H),6.77(d,J=5.9Hz,1H),4.01(s ,3H), 2.58(s,3H).

(2) Preparation of N-(2-chloropyrimidin-4-yl)-N,2,3-trimethyl-2H-indazol-6-amine (3)

Add 1.00g of product 2 from the previous step and 2.40g of cesium carbonate to 50mL N,N-dimethylformamide, react at room temperature for 20min, then slowly add 0.78g of methyl iodide, after the addition is complete, react at room temperature for 2h, pour the reaction solution into A large amount of light yellow solid was precipitated immediately in ice water, filtered, dried, and recrystallized with ethyl acetate to obtain 1.06 g of product 3, which was light yellow crystal, yield: 80.0%; Mp: 173-174 ° C. ¹ H NMR ( 400MHz, DMSO-d ₆ )δ7.95(d, J=6.1Hz,1H),7.80(dd,J=0.8,8.7Hz,1H),7.51(dd,J=0.8,1.8Hz,1H),6.88 (dd, J=1.8, 8.8Hz, 1H), 6.24(d, J=6.1Hz, 1H), 4.06(s, 3H), 3.42(s, 3H), 2.63(s, 3H).

(3) Preparation of 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoic acid (4a)

Disperse 0.50g of intermediate 3 and 0.36g of p-aminobenzoic acid in 30mL of isopropanol, add 2 drops of concentrated hydrochloric acid, heat to 85°C, reflux for 4h, cool to room temperature, filter, and wash with isopropanol and ethyl acetate The filter cake was dried to obtain 0.53g product 4a as a white solid, yield: 80%; Mp:>250°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ12.16(s,1H), 9.80(s, 1H), 7.91(d, J=6.3Hz, 1H), 7.79(d, J=8.7Hz, 3H), 7.74(d, J=8.6Hz, 2H), 7.49(d, J=1.7Hz, 1H) ,6.91(dd,J=1.8,8.8Hz,1H),5.94(d,J=6.2Hz,1H),4.08(s,3H),3.51(s,3H),2.64(s,3H).ESI- MS m/z: 387.5 [MH] ^- .

(4) Methyl 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoate (5a ) preparation

Add 0.50g of 4a to 50mL of anhydrous methanol, then add 0.60g of thionyl chloride dropwise at 0°C, after the addition is complete, keep warm for 30min, remove the ice bath and heat to reflux, react for 5h, and depressurize the solvent After distillation, the crude product was recrystallized with isopropanol to obtain 0.50 g of product 5a as a white solid, yield: 96%; Mp: 198-200°C. ¹ H NMR (400MHz, DMSO-d ₆ )δ9.63(s, 1H), 7.92(d, J=6.0Hz, 1H), 7.83(d, J=8.6Hz, 2H), 7.78(d, J= 8.6Hz, 1H), 7.72(d, J=8.6Hz, 2H), 7.46(dd, J=0.8, 1.7Hz, 1H), 6.89(dd, J=1.8, 8.8Hz, 1H), 5.91(d, J＝6.0Hz,1H),4.07(s,3H),3.80(s,3H),3.49(s,3H),2.64(s,3H).ESI-MS m/z:403.5[M+H] ⁺ .

(5) 4-((4-((2,3-dimethyl-2H-indazol-6-yl) (methyl) amino) pyrimidin-2-yl) amino)-N-hydroxybenzamide ( 6a) Preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Compound 5a 0.20g was dissolved in 30mL potassium hydroxylamine solution, stirred at room temperature for 2h, concentrated under reduced pressure to remove most of the solvent, the remaining residue was adjusted to pH 5-6 with 1M HCl, a white solid was precipitated, filtered, and the filter cake was dried to obtain 80.6mg Product 6a, white solid, yield: 40%; Mp: 210-212°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.99(s, 1H), 9.49(s, 1H), 8.85(s ,1H),7.89(d,J=6.1Hz,1H),7.84–7.70(m,3H),7.59(d,J=8.3Hz,2H),7.46(s,1H),6.90(d,J= 8.7Hz,1H),5.86(d,J=6.2Hz,1H),4.07(s,3H),3.49(s,3H),2.64(s,3H).HRMS(AP-ESI)m/z calcd for C ₂₁ H ₂₁ N ₇ O ₂ [M+H] ⁺ 404.1757, found 404.1739. Embodiment 2: the synthesis of compound 6d-6e, taking 6d as example

(1) N-(2-aminophenyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl) Preparation of amino) benzamide (6d)

Add 0.50 g of intermediate 4a to 50 mL of anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethylurea Fluoroboric acid 0.49g and triethylamine 0.26g, activated for 30 minutes, then add o-phenylenediamine 0.17g, react overnight at room temperature, add ethyl acetate to dilute after the reaction is completed, wash to remove N,N-dimethylformamide , the ethyl acetate layer was washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=30:1 volume ratio eluent 0.40 g of product 6d was obtained as an off-white solid, yield: 65%; Mp: 138-140° C. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.54 (s, 1H), 9.49 (s, 1H), 7.91(d,J=6.0Hz,1H),7.87(d,J=9.0Hz,2H),7.83(d,J=8.8Hz,2H),7.80-7.76(m,1H),7.47(d,J =2.3Hz,1H),7.17(dd,J=1.5,7.9Hz,1H),6.96(td,J=1.6,7.6Hz,1H),6.91(dd,J=1.8,8.8Hz,1H),6.79 (dd, J=1.5,8.0Hz,1H),6.60(td,J=1.5,7.5Hz,1H),5.87(d,J=6.0Hz,1H),4.89(s,2H),4.06(s, 3H), 3.51(s,3H), 2.63(s,3H). HRMS (AP-ESI) m/z calcd for C ₂₁ H ₂₁ N ₇ O ₂ [M+H] ⁺ 479.2230, found 479.2257.

Embodiment 3: the synthesis of compound 10a-10b, taking 10a as example

(1) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-methoxy-N -Preparation of methylbenzamide (7a)

Add 1.00 g of intermediate 4a to anhydrous N,N-dimethylformamide, under ice-cooling, add O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoro 0.98g of boric acid and 0.52g of triethylamine, activated for 30min, then added 0.30g of N,O-dimethyl hydroxylamine hydrochloride and 0.31g of triethylamine, reacted at room temperature overnight, after the reaction was completed, diluted with ethyl acetate, washed with water to remove Remove N,N-dimethylformamide, wash the ethyl acetate layer with saturated sodium bicarbonate solution and saturated NaCl solution, dry over anhydrous magnesium sulfate, remove the solvent under reduced pressure, and wash the residue with dichloromethane:methanol=35:1 volume Purified by column chromatography with a specific eluent to obtain 0.78g of product 7a as a white solid with a yield of 70%; Mp: 216-218°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.46(s, 1H), 7.90(d, J=6.0Hz, 1H), 7.82-7.74(m, 3H), 7.48(d, J=8.6Hz, 2H), 7.46(d, J=1.4Hz, 1H), 6.89( dd,J=1.8,8.8Hz,1H),5.87(d,J=6.0Hz,1H),4.06(s,3H),3.55(s,3H),3.48(s,3H),3.23(s,3H ), 2.63(s,3H).

(2) Preparation of 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzaldehyde (8a)

1.00 g of compound 7a was dissolved in 50 mL of anhydrous tetrahydrofuran, and cooled to -40 °C in a cryogenic reactor. Slowly add 0.26 g of lithium aluminum tetrahydrogen to it in batches, and maintain the temperature for 2 hours. After the reaction is completed, slowly and carefully quench the reaction with ice water until no more bubbles are generated. The reaction solution was diluted with water, extracted with ethyl acetate, the organic layers were combined, washed with saturated NaCl solution, dried over anhydrous magnesium sulfate, the solvent was concentrated under reduced pressure, and the residue was subjected to column separation with dichloromethane:methanol=40:1 volume ratio eluent Purified by chromatography to obtain 0.61 g of product 8a as a white solid, yield: 70%; Mp: 242-244°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.77 (d, J=4.6Hz, 2H) ,7.96-7.93(m,3H),7.78(d,J=8.7Hz,1H),7.69(d,J=8.4Hz,2H),7.47(d,J=1.6Hz,1H),6.90(dd, J=1.8, 8.8Hz, 1H), 5.92(d, J=6.0Hz, 1H), 4.07(s, 3H), 3.50(s, 3H), 2.64(s, 3H).

(3) Ethyl (E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino ) Preparation of phenyl) acrylate (9a)

Add 0.32 g of triethyl phosphoroacetate to 50 mL of anhydrous tetrahydrofuran, cool to -5-0°C, slowly add 0.16 g of NaH, after the addition is complete, stir for 20 min, then add 0.52 g of 8a in tetrahydrofuran dropwise, react overnight at room temperature, After the reaction was completed, 50 mL of 10% ammonium chloride solution was added, stirred for 30 min, and then separated into layers. After the organic phase was dried, filtered, and concentrated to obtain a crude product. The residue was purified by column chromatography with dichloromethane:methanol=40:1 volume ratio eluent to obtain 0.48g product 9a as a white solid, yield: 48%; Mp: 195-197°C. ¹ H NMR (400MHz ,DMSO-d ₆ )δ9.50(s,1H),7.89(d,J=6.0Hz,1H),7.83(d,J=8.7Hz,2H),7.78(dd,J=0.8,8.7Hz, 1H), 7.58–7.51(m, 3H), 7.46(dd, J＝0.8, 1.8Hz, 1H), 6.90(dd, J＝1.8, 8.8Hz, 1H), 6.42(d, J＝16.0Hz, 1H ),5.84(d,J=6.0Hz,1H),4.17(q,J=7.1Hz,2H),4.07(s,3H),3.49(s,3H),2.64(s,3H),1.25(t , J=7.1Hz, 3H). (4) (E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene base)-N-hydroxyacrylamide (10a) preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.50 g of compound 9a in 30 mL of potassium hydroxylamine solution, stir at room temperature for 2 h, then concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 0.29 g Product 10a, a white solid, yield: 60%; Mp: 194-196°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.65(s, 1H), 9.42(s, 1H), 8.96(s ,1H),7.88(d,J=5.9Hz,1H),7.81(d,J=8.3Hz,2H),7.77(d,J=8.8Hz,1H),7.46(s,1H),7.43-7.32 (m,3H),6.95-6.83(m,1H),6.28(d,J=15.7Hz,1H),5.83(d,J=6.1Hz,1H),4.07(s,3H),3.49(s, 3H), 2.64(s,3H). HRMS (AP-ESI) m/z calcd for C ₂₃ H ₂₃ N ₇ O ₂ [M+H] ⁺ 430.1991, found 430.1988.

Embodiment 4: the synthesis of compound 13a-13i, taking 13a as an example

(1) Methyl (4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoyl)glycinate (11a) preparation of

Add 1.00 g of intermediate 4a to 100 mL of anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethylurea Fluoroboric acid 0.98g and triethylamine 0.52g, activated for 30 minutes, then added glycine methyl ester hydrochloride 0.39g, reacted overnight at room temperature, after the reaction was completed, added ethyl acetate to dilute, washed with water to remove N,N-dimethyl The formamide and ethyl acetate layers were washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure, and the residue was subjected to column chromatography with dichloromethane:methanol=30:1 volume ratio Analysis and purification yielded 0.87g of product 11a as a white solid with a yield of 74%; Mp: 170-172°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.51(s, 1H), 8.72(t, J =5.9Hz, 1H), 7.90(d, J=6.0Hz, 1H), 7.84(d, J=8.6Hz, 2H), 7.77(d, J=8.7Hz, 1H), 7.71(d, J=8.5 Hz,2H),7.46(d,J=1.7Hz,1H),6.90(dd,J=1.8,8.8Hz,1H),5.85(d,J=6.1Hz,1H),4.07(s,3H), 3.98 (d, J=5.8Hz, 2H), 3.65 (s, 3H), 3.50 (s, 3H), 2.64 (s, 3H).

(2) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(2-(hydroxylamine )-2-oxoethyl) benzamide (13a) preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Compound 11a (0.50g, 1.09mmol) was dissolved in 30mL potassium hydroxylamine solution, stirred at room temperature for 2h, concentrated under reduced pressure to remove most of the solvent, the remaining residue was adjusted to pH 5-6 with 1M HCl, a white solid was precipitated, filtered, dried and filtered Cake, the product 13a was obtained as a white solid (0.31 g, yield 61%), Mp: 216-218 °C. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ10.53 (s, 1H), 9.46 (s ,1H),8.77(s,1H),8.45(s,1H),7.89(d,J=5.9Hz,1H),7.82(d,J=8.5Hz,2H),7.77(d,J=8.7Hz ,1H),7.72(d,J=8.4Hz,2H),7.46(d,J=1.8Hz,1H),6.90(dd,J=1.8,8.8Hz,1H),5.85(d,J=6.0Hz ₂₃ H ₂₄ N ₈ O ₃ [MH] ^- 459.1971, found 459.1921.

Embodiment 5: the synthesis of compound 14a-14g, taking 14a as an example

(1) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoyl)glycine (12a) preparation of

11a (1.00g, 2.18mmol) was added to 30mL of methanol, then 5mL of 3M NaOH solution was added, and stirred at room temperature for 4h. After the reaction was completed, most of the methanol was evaporated under reduced pressure, and the residue was adjusted to pH 5- 6. After filtration, the filter cake was washed with ice water and dried to obtain the product 12a as a white solid (0.81g, yield: 84%), Mp:>250°C, directly submitted to the next step. ¹ H NMR (400MHz, DMSO-d ₆ ) δ12.57(s,1H),10.69(s,1H),8.79(s,1H),8.01–7.61(m,6H),7.59(d,J=1.7Hz,1H),6.95(dd,J= 1.8, 8.8Hz, 1H), 6.03(s, 1H), 4.09(s, 3H), 3.92(d, J=5.8Hz, 2H), 3.57(s, 3H), 2.66(s, 3H).

(2) N-(2-((2-aminophenyl)amino)-2-oxoethyl)-4-((4-((2,3-dimethyl-2H-indazole-6- base) (methyl) amino) pyrimidin-2-yl) amino) benzamide (14a) preparation

Add intermediate 12a (1g, 2.24mmol) to anhydrous N,N-dimethylformamide, under ice-cooling, add O-benzotriazole-N,N,N',N'-tetramethyl Urea tetrafluoroboric acid (0.86g, 2.69mmol) and triethylamine (0.45g, 4.48mmol), activated for 30min, then added o-phenylenediamine (0.29g, 2.69mmol), reacted overnight at room temperature, after the reaction was completed Add ethyl acetate for dilution, wash with water to remove N,N-dimethylformamide, wash the ethyl acetate layer with saturated sodium bicarbonate solution and saturated NaCl solution, dry over anhydrous magnesium sulfate, remove the solvent under reduced pressure, and wash the residue with dichloro Methane: Methanol = 30:1 volume ratio of eluent for column chromatography purification to obtain product 14a, light yellow solid (0.47g, yield 39%), Mp: 178-180 ° C. ¹ H NMR (400MHz, DMSO -d ₆ )δ9.51(s,1H),9.23(s,1H),8.62(t,J＝5.8Hz,1H),7.90(d,J＝6.1Hz,1H),7.83(d,J＝ 8.7Hz, 2H), 7.79-7.71(m, 3H), 7.49-7.43(m, 1H), 7.12(d, J=7.5Hz, 1H), 6.93-6.89(m, 2H), 6.71(dd, J =1.4,7.9Hz,1H),6.54(t,J=7.5Hz,1H),5.86(d,J=6.2Hz,1H),4.92(s,2H),4.07(s,3H),4.05-4.00 (m,2H), 3.50(s,3H), 2.64(s,3H). HRMS (AP-ESI) m/z calcd for C ₂₉ H ₂₉ N ₉ O ₂ [M+H] ⁺ 536.2444, found 536.2498.

Embodiment 6: the synthesis of compound 19a-19g, taking 19f as an example

(1) Preparation of methyl 7-(4-nitrophenoxy)heptanoate (16f)

Add 1.00g of p-nitrophenol, 1.93g of methyl 7-bromoheptanoate and 2.98g of potassium carbonate to 50mL of acetone, raise the temperature to reflux, and react overnight. Crystallization afforded 1.62 g of product 16f as a yellow solid, yield: 80%; ESI-MS m/z: 282.3 [M+H] ⁺ .

(2) Preparation of methyl 7-(4-aminophenoxy)heptanoate (17f)

Add 1.00 g of intermediate 16f to 50 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, react overnight at room temperature, filter, wash the filter residue with methanol, combine the filtrate and washing liquid, and evaporate under reduced pressure After removing the solvent, 0.79 g of the product 17f was obtained as a light brown solid, yield: 88%; ESI-MS m/z: 251.4[M+H] ⁺ , which was directly put into the next step.

(3) Methyl 7-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy )-Heptanoic acid ester (18f) preparation

Add 0.5 g of intermediate 3, 0.52 g of intermediate 17f and 0.5 mL of concentrated hydrochloric acid into 50 mL of isopropanol at room temperature, raise the temperature to reflux, and react overnight. After the reaction, the solvent was evaporated under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=40:1 volume ratio eluent to obtain 0.61g of product 18f as a white solid, yield: 70%; ESI - MS m/z: 503.6 [M+H] ⁺ .

(4) 7-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)- Preparation of N-Hydroxyheptanamide (19f)

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.30 g of compound 18f in 30 mL potassium hydroxylamine solution, stir at room temperature for 2 h, concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 0.20 g Product 19f, white solid, yield: 65%; ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.33(s, 1H), 8.93(s, 1H), 8.65(d, J=1.8Hz, 1H ),7.80(d,J=6.0Hz,1H),7.75(dd,J=0.8,8.7Hz,1H),7.60(d,J=8.9Hz,2H),7.42(dd,J=0.8,1.8Hz ,1H),6.87(dd,J=1.8,8.8Hz,1H),6.75(d,J=8.9Hz,2H),5.73(d,J=5.9Hz,1H),4.06(s,3H),3.88 (t,J=6.5Hz,2H),3.45(s,3H),2.63(s,3H),2.02-1.90(m,2H),1.67(p,J=6.8Hz,2H),1.51(p, J = 7.4 Hz, 2H), 1.39 (p, J = 7.2 Hz, 2H), 1.34-1.21 (m, 2H). ESI-MS m/z: 504.7 [M+H] ⁺ .

Embodiment 7: the synthesis of compound 23a-23g, taking compound 23f as an example

(1) N ⁴ -(2,3-dimethyl-2H-indazol-6-yl)-N ⁴ -methyl-N ² -(4-nitrophenyl)pyrimidine-2,4-diamine Preparation of (20)

Add 1.00g of intermediate 3, 0.58g of p-nitroaniline and 0.5mL of concentrated hydrochloric acid to 70mL of isopropanol, raise the temperature to reflux, and react overnight. After the reaction, cool to room temperature, filter, and filter the cake with a small amount of isopropanol Wash and dry the filter cake to obtain 1.08g product 20 as light yellow solid, yield: 80%; ¹ H NMR (400MHz, DMSO-d ₆ ) δ11.01(s, 1H), 8.08(s, 2H), 7.99(d, J=7.0Hz, 1H), 7.87(d, J=8.8Hz, 1H), 7.83(s, 2H), 7.59(d, J=1.8Hz, 1H), 6.95(dd, J=1.8 ,8.7Hz,1H),6.17(s,1H),4.10(s,3H),3.58(s,3H),2.67(s,3H).

(2) N ² -(4-aminophenyl)-N ⁴ -(2,3-dimethyl-2H-indazol-6-yl)-N ⁴ -methylpyrimidine-2,4-diamine ( 21) Preparation

Suspend 1.00 g of intermediate 20 in 70 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, and react overnight at room temperature. After the reaction is completed, the solvent is evaporated under reduced pressure, and the residue is recrystallized with ethyl acetate 0.78 g of product 21 was obtained as a light brown solid, yield: 84%; ESI-MS m/z: 360.4 [M+H] ⁺ .

(3) Methyl 8-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl )amino)-8-oxooctyl ester (22f) preparation

Add 0.33g of monomethyl suberate to 50mL of anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethyl After adding 0.68g of urea tetrafluoroboric acid and 0.36g of triethylamine, activate in ice bath for 30min. After activation, add 0.76g of intermediate 21, remove the ice bath, and react overnight at room temperature. After the reaction is completed, add ethyl acetate to dilute. Wash with water to remove N,N-dimethylformamide, wash the ethyl acetate layer with saturated sodium bicarbonate solution and saturated NaCl solution, dry over anhydrous magnesium sulfate, evaporate the solvent under reduced pressure, and dichloromethane:methanol=30 : 1 volume ratio eluent was purified by column chromatography to obtain 0.37g product 22f as a white solid, yield: 40%; ESI-MS m/z: 530.5[M+H] ⁺ .

(4) N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)- Preparation of N ⁸ -Hydroxysuberamide (23f)

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.20 g of compound 22f in 30 mL potassium hydroxylamine solution, stir at room temperature for 2 h, concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 0.14 g Product 23f, white solid, yield: 70%; ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.34(s, 1H), 9.68(s, 1H), 9.04(s, 1H), 8.65(s ,1H),7.82(d,J=5.9Hz,1H),7.75(d,J=8.8Hz,1H),7.64–7.56(m,2H),7.43(d,J=1.7Hz,1H),7.37 (d,J=8.9Hz,2H),6.88(dd,J=1.8,8.8Hz,1H),5.76(d,J=6.0Hz,1H),4.06(s,3H),3.46(s,3H) ,2.63(s,3H),2.25(t,J=7.4Hz,2H),1.94(t,J=7.4Hz,2H),1.64-1.40(m,4H),1.34-1.21(m,4H). ESI-MS m/z: 531.6 [M+H] ⁺ .

Embodiment 8: the synthesis of compound 28a-28g, taking compound 28e as an example

(1) Preparation of methyl 6-((4-nitrophenyl)amino)hexyl ester (25e)

Add 1.00g of p-fluoronitrobenzene, 1.54g of methyl 6-aminocaproate hydrochloride and 2.94g of potassium carbonate into 50mL of N,N-dimethylformamide, raise the temperature to 50°C overnight, and after the reaction , cooled to room temperature, the reaction solution was poured into ice water, a yellow solid was precipitated, filtered, dried, and the residue was subjected to column chromatography with petroleum ether:ethyl acetate=4:1 volume ratio eluent to obtain 1.32g of product 25e, It is light yellow solid, yield: 70%; ESI-MS m/z: 267.3[M+H] ⁺ .

(2) Preparation of methyl 6-((4-aminophenyl)amino)hexyl ester (26e)

Suspend 1.00 g of intermediate 25e in 70 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, and react overnight at room temperature. After the reaction is completed, the solvent is evaporated under reduced pressure, and the residue is recrystallized with ethyl acetate 0.71 g of product 26e was obtained as a brown solid, yield: 80%; ESI-MS m/z: 237.4 [M+H] ⁺ .

(3) Methyl 6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl )amino)-hexyl ester (27e) preparation

Add 0.5 g of intermediate 3, 0.49 g of intermediate 26e and 0.5 mL of concentrated hydrochloric acid into 50 mL of isopropanol at room temperature, raise the temperature to reflux, and react overnight. After the reaction was completed, the solvent was evaporated under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=40:1 volume ratio eluent to obtain product 27e, which was 0.55g of light yellow solid, yield: 65%; ESI - MS m/z: 488.6[M+H] ⁺ .

(4) 6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino )-N-hydroxy caproamide (28e) preparation

28.0 g of KOH and 23.53 g of hydroxylamine hydrochloride were dissolved in 70 mL and 120 mL of anhydrous methanol respectively to obtain solution A and solution B. Under an ice bath, solution A was added dropwise to solution B, and after stirring for 2 hours, the precipitate was filtered off, and the obtained filtrate was a methanol solution of potassium hydroxylamine. Dissolve 0.20 g of compound 27e in 30 mL of potassium hydroxylamine solution, stir at room temperature for 2 h, then concentrate under reduced pressure to remove most of the solvent, and adjust the pH of the remaining residue to 5-6 with 1M HCl, precipitate a white solid, filter, and dry the filter cake to obtain 0.13 g Product 28e, as a reddish solid, yield: 66%. ¹ H NMR (400MHz,DMSO-d ₆ )δ10.36(s,1H),8.78(s,1H),8.68(s,1H),7.76-7.73(m,2H),7.43(d,J=1.6 Hz,1H),7.37(d,J=8.4Hz,2H),6.87(dd,J=1.8,8.8Hz,1H),6.45(d,J=8.4Hz,2H),5.68(d,J=6.0 Hz,1H),4.06(s,3H),3.49(s,1H),3.44(s,3H),2.93(t,J=7.0Hz,2H),1.96(t,J=7.3Hz,2H), 1.61-1.10 (m, 6H). ESI-MS m/z: 489.3 [M+H] ⁺ .

Embodiment 9: In vitro HDAC inhibitory activity test experiment of compound

We first used the readily available HeLa nuclear extract (mainly containing HDAC1 and HDAC2) as the source of HDAC enzymes, and determined the HDACs inhibitory activity of the target compounds in vitro.

In vitro HDAC inhibitory activity of the compounds in Table 1

Experimental conclusion: Most of the compounds have a certain inhibitory effect on HDACs, and the activities of 13e and 13f among the hydroxamic acid compounds are the best, which are obviously better than the positive control drug SAHA. Among o-phenylenediamine compounds, 6d and 14e had the best activity, which was equivalent to the positive control drug MS275.

Example 10: In vitro HDAC subtype selectivity experiment of representative compounds

Table 2 In vitro HDAC subtype selectivity of representative compounds

^a not determined

Experimental conclusion: The representative compounds 10a and 13f of hydroxamic acids are similar to the positive control drug SAHA, both of which are broad-spectrum HDAC inhibitors, but the inhibitory activity of compound 13f on HDAC6 is better than that of SAHA. Representative o-phenylenediamine compounds 6d and 14g are similar to the positive control drug MS275, and both are Class I selective HDAC inhibitors.

Example 11: In vitro VEGFR-2 inhibitory activity assay of compounds

Firstly, the percent inhibition rate of all target compounds on VEGFR-2 at the concentration of 0.2 μM was determined, and the in vitro anti-VEGFR-2 IC ₅₀ values of four representative compounds (10a, 13f, 6d, 14g) were further determined.

The in vitro VEGFR-2 inhibitory activity of the compound of table 3

^a not determined

Experimental conclusion: Most of the compounds showed strong VEGFR-2 inhibitory activity at 200nM (inhibition rate>90%), representative compounds 10a, 13f, 6d and 14g all showed the same in vitro VEGFR activity as the positive control drug Pazapanib -2 inhibitory activity.

Example 12: Western blot experiments of representative compounds

In order to further study the inhibitory effect of compounds on HDAC in cells, we conducted Western Blot experiments to measure the effects of representative compounds 6d and 13f on the expression levels of acetylated H4 (Class I HDAC substrate) and acetylated tubulin (HDAC6 substrate). Among them, SAHA, TubA and MS275 are positive control drugs, and β-Actin is an internal standard. See Figure 1 for the experimental results.

Experimental conclusion: compound 6d can significantly increase the level of acetylated histone H4 (Ac-HH4) at 1 μM, and its activity is equivalent to that of MS275, but compound 6d cannot increase the level of acetylated tubulin (Ac-tubulin), indicating that compound 6d is A Class I HDAC selective inhibitor. Compound 13f can significantly increase the level of acetylated tubulin (Ac-tubulin) at 100nM, and its activity is better than that of TubA and SAHA; 13f can also increase the level of acetylated histone H4 (Ac-HH4), indicating that compound 13f is a broad-spectrum HDAC inhibitors.

Example 13: In vitro anti-tumor cell proliferation activity experiment of representative compounds

We used the commonly used MTT method to measure the in vitro anti-tumor cell proliferation activity of the test compounds.

The in vitro anti-tumor cell proliferation activity of table 4 representative compounds

^a not determined

Experimental conclusion: Most of the compounds showed certain anti-proliferation effects on a variety of hematological tumors and solid tumor cell lines, among which compounds 10a and 13f had the best activity among the hydroxamic acid compounds, showing a similar effect to positive control drugs. SAHA comparable anti-tumor cell proliferation activity. Among the o-phenylenediamine compounds, compound 6d has the best activity, showing the same anti-tumor cell proliferation activity as the positive control drug MS275.

Example 14 Representative Compounds Inhibit Human Umbilical Vein Endothelial Cells Angiogenesis Lumen Experiment

We selected representative compounds 6d and 13f to measure the effect of the compounds on the lumen formation of HUVECs at a concentration of 1 μM, and selected Pazopanib as a positive control drug. See Figure 2 for the experimental results.

Experimental conclusions: Representative compounds 6d and 13f exhibited comparable in vitro anti-HUVECs angiogenesis lumen activity to that of the positive control drug Pazopanib at a concentration of 1 μM.

Rat thoracic aorta angiogenesis inhibitory activity experiment of representative compound of embodiment 15

We further conducted rat thoracic aorta angiogenesis experiments to further evaluate the in vitro anti-angiogenic activity of the compounds, and determined the effects of representative compounds 6d and 13f on rat thoracic aorta angiogenesis at a concentration of 5 μM, and the positive control drug was Pazopanib . See Figure 3 for the experimental results.

Experimental conclusion: Compounds 6d, 13f and Pazopanib can almost completely inhibit the formation of arterial ring microvessels at a concentration of 5 μM.

Antitumor activity experiment in vivo of embodiment 16 representative compound

Based on the excellent in vitro anti-tumor cell proliferation activity of compounds 6d and 13f in HT-29 cell line, we established a subcutaneous HT-29 tumor-bearing model in nude mice, using SAHA and Pazopanib as positive controls. Compounds 6d and 13f were orally administered at two doses of 50 mg/kg/day and 100 mg/kg/day, SAHA was orally administered at 100 mg/kg/day and Pazopanib was orally administered at 50 mg/kg/day. After 21 days of administration, the animals were sacrificed to end the experiment, the tumor growth inhibition rate (TGI) of the test compound and the positive control drug was calculated, and the tumor inhibitory activity of the compound was evaluated from two aspects of tumor volume and weight. See Figure 4 and Figure 5 for the experimental results.

Experimental conclusion: Compounds 6d and 13f both showed anticancer activity in vivo, and 6d had better activity. At a dose of 100mg/kg/day, the 6d activity is better than that of the marketed HDAC inhibitor drug SAHA, and at a dose of 50mg/kg/day, the 6d activity is comparable to that of the marketed VEGFR inhibitor drug Pazopanib, with a very high potency Follow-up research and development value.

Claims (10)

1. HDAC and VEGFR dual target inhibitors based on pazopanib structure, its pharmaceutically acceptable salt, solvate or prodrug, is characterized in that, has the structure shown in following general formula I, II or III : in X is n is 0-9; Y is 2. the HDAC and VEGFR dual-target inhibitor based on pazopanib structure as claimed in claim 1, is characterized in that, Y in general formula I and X in II are in the para-position or meta-position of the benzene ring X is n is 0-7; Y is 3. The HDAC and VEGFR dual-target inhibitor based on the pazopanib structure according to claim 1 or 2, wherein the compound is one of the following: 4. The preparation method of the HDAC and VEGFR dual-target inhibitor based on the pazopanib structure as claimed in claim 1 is one of the following methods: (1) Using 2,3-dimethyl-6-amino-2H-indazole as the starting material, it undergoes a nucleophilic substitution reaction with 2,4-dichloropyrimidine under sodium bicarbonate alkaline conditions to obtain intermediate 2 , intermediate 2 reacts with methyl iodide under the catalysis of cesium carbonate to obtain intermediate 3, and intermediate 3 undergoes nucleophilic substitution reaction with different substituted anilines under the catalysis of concentrated hydrochloric acid to obtain key intermediate 4. Intermediate 4 is protected by methyl ester Intermediate 5 is obtained, and intermediate 5 is subjected to ester aminolysis reaction to obtain final product 6a-6c; in addition, intermediate 4a-4b is condensed with o-phenylenediamine to obtain final product 6d-6e; The reaction formula is as follows: Among them, the benzene rings of 4a, 5a, 6a, and 6d are para-substituted, and the benzene rings of 4b, 5b, 6b, and 6e are meta-substituted; Reagents and conditions in the above reaction formula: (a) 2,4-dichloropyrimidine, sodium bicarbonate, absolute ethanol, reflux, 4h; (b) cesium carbonate, methyl iodide, N,N-dimethylformamide , room temperature, 2h; (c) isopropanol, concentrated hydrochloric acid, reflux, 4h; (d) O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, triethyl Amine, o-phenylenediamine, N,N-dimethylformamide, 0°C-room temperature, overnight (e) anhydrous methanol, thionyl chloride, reflux, overnight; (f) hydroxylamine hydrochloride, potassium hydroxide, no Water methanol, room temperature, 2h; The different substituted anilines are: p-aminobenzoic acid, m-aminobenzoic acid or 5-amino-2-methylbenzoic acid; (2) Using intermediate 4 as the starting material, it undergoes amide condensation with N,O-dimethyl hydroxylamine hydrochloride to obtain intermediate 7, which is reduced to aldehyde intermediate 8 by tetrahydrolithium aluminum, and intermediate 8 is combined with phosphorus Triethyl acyl acetate undergoes Horner-Wadsworth-Emmons reaction to obtain intermediate 9, and intermediate 9 is subjected to ester aminolysis to obtain final product 10; The reaction formula is as follows: Among them, the benzene rings of 4a, 7a, 8a, 9a, and 10a are para-substituted, and the benzene rings of 4b, 7b, 8b, 9b, and 10b are meta-substituted; Reagents and conditions in the above reaction formula: (a) N,O-dimethylhydroxylamine hydrochloride, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethyl Amine, N,N-dimethylformamide, room temperature, overnight; (b) lithium aluminum tetrahydrogen, tetrahydrofuran, -20°C, 4h; (c) triethyl phosphoroacetate, sodium hydride, tetrahydrofuran, 0°C- Room temperature, overnight; (d) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h; (3) Using intermediate 4 as the starting material, intermediate 4 and aminoalkane methyl esters of different chain lengths undergo amide condensation reaction to obtain intermediate 11, and intermediate 11 obtains final product 13 through the aminolysis reaction of ester; intermediate 11 is hydrolyzed into intermediate 12 under alkaline conditions of sodium hydroxide, and intermediate 12 and o-phenylenediamine undergo amide condensation reaction to obtain final product 14; The reaction formula is as follows: Among them, the benzene rings of 4a, 11a-11g, 12a-12g, 13a-13g, and 14a-14g are para-substituted, and the benzene rings of 4b, 11h, and 13h are meta-substituted, and n is 1-7; Reagents and conditions in the above reaction formula: (a) aminoalkane methyl esters with different chain lengths, O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, triethyl Amine, N,N-dimethylformamide, room temperature, overnight; (b) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h; (c) 3M NaOH, methanol, room temperature, 2h; (d) o Phenylenediamine, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, N,N-dimethylformamide, room temperature, overnight; The aminoalkane methyl esters of different chain lengths are: glycine methyl ester, 3-aminopropionic acid methyl ester, 4-aminobutyric acid methyl ester, 5-aminovaleric acid methyl ester, 6-aminocaproic acid methyl ester, Methyl 7-aminoheptanoate or methyl 8-aminooctanoate; (4) Using p-nitrophenol as the starting material, nucleophilic substitution reaction occurs with methyl bromoalkanoate of different chain lengths to obtain intermediate 16, intermediate 16 is reduced by Pd/C to obtain intermediate 17, intermediate 17 and 3. A nucleophilic reaction occurs to obtain intermediate 18, and intermediate 18 undergoes ester ammonolysis to obtain final product 19; The reaction formula is as follows: Among them, n is 1-7; Reagents and conditions in the above reaction formula: (a) methyl bromoalkanoate with different chain lengths, potassium carbonate, acetone, reflux, overnight; (b) Pd/C, methanol, hydrogen, room temperature, overnight; (c) isopropyl Alcohol, concentrated hydrochloric acid, reflux, overnight; (d) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h; The methyl bromoalkanoates of different chain lengths are: methyl 2-bromoacetate, methyl 3-bromopropionate, methyl 4-bromobutyrate, methyl 5-bromovalerate, 6-bromohexanoic acid Methyl ester, methyl 7-bromoheptanoate or methyl 8-bromooctanoate; (5) Using intermediate 3 as the starting material, nucleophilic substitution reaction occurs with p-nitroaniline to obtain intermediate 20. Intermediate 20 is reduced to intermediate 21 by Pd/C. Acid monomethyl ester undergoes amide condensation to obtain intermediate 22, and intermediate 22 undergoes ester ammonolysis to obtain final product 23; The reaction formula is as follows: Among them, n is 1-7; Reagents and conditions in the above reaction formula: (a) p-nitroaniline, isopropanol, concentrated hydrochloric acid, reflux, overnight; (b) Pd/C, methanol, hydrogen, room temperature, overnight; (c) different chain lengths Monomethyl alkanedioate, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, N'N-dimethylformamide, room temperature, overnight ; (d) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h; The monomethyl alkanedioic acid of described different chain lengths is: monomethyl malonate, monomethyl succinate, monomethyl glutarate, monomethyl adipate, monomethyl pimelate, Monomethyl suberate or monomethyl azelate; (6) Using p-fluoronitrobenzene as the starting material, nucleophilic substitution reaction with aminoalkanoic acid methyl esters of different chain lengths produces intermediate 25, intermediate 25 is reduced by Pd/C to obtain intermediate 26, intermediate 26 Nucleophilic substitution reaction with intermediate 3 to obtain intermediate 27, and intermediate 27 to obtain intermediate 28 through ester ammonolysis; The reaction formula is as follows: Among them, n is 1-7; Reagents and conditions in the above reaction formula: (a) methyl aminoalkanoates with different chain lengths, potassium carbonate, N,N-dimethylformamide, 50°C, overnight; (b) Pd/C, hydrogen, methanol , overnight; (c) concentrated hydrochloric acid, isopropanol, reflux, overnight; (d) hydroxylamine hydrochloride, potassium hydroxide, anhydrous methanol, room temperature, 2h; The aminoalkane methyl esters of different chain lengths are: glycine methyl ester, 3-aminopropionic acid methyl ester, 4-aminobutyric acid methyl ester, 5-aminovaleric acid methyl ester, 6-aminocaproic acid methyl ester, Methyl 7-aminoheptanoate or methyl 8-aminooctanoate. 5. The preparation method of HDAC and VEGFR dual-target inhibitor based on pazopanib structure as claimed in claim 4 is one of the following methods: (1) 4-((4-((2,3-dimethyl-2H-indazol-6-yl) (methyl) amino) pyrimidin-2-yl) amino)-N-hydroxybenzamide ( 6a) preparation method, the steps are as follows: (1) Preparation of N-(2-chloropyrimidin-4-yl)-2,3-dimethyl-6-amino-2H-indazole (2) Dissolve 5.00g of 2,3-dimethyl-6-amino-2H-indazole, 13.9g of 2,4-dichloropyrimidine and 10.4g of sodium bicarbonate in 100mL of absolute ethanol, heat to 79°C, and reflux After 4 hours, cool down to room temperature, filter, and wash the filter cake fully with ethyl acetate, collect the filtrate, concentrate under reduced pressure, remove the solvent, fully beat with ethyl acetate, filter to obtain an off-white solid, and recrystallize with methanol to obtain 7.64g of the product 2. It is a white solid; (2) Preparation of N-(2-chloropyrimidin-4-yl)-N,2,3-trimethyl-2H-indazol-6-amine (3) Add 1.00g of product 2 from the previous step and 2.40g of cesium carbonate to 50mL N,N-dimethylformamide, react at room temperature for 20min, then slowly add 0.78g of methyl iodide, after the addition is complete, react at room temperature for 2h, pour the reaction solution into A large amount of light yellow solid precipitated immediately in ice water, filtered, dried, and recrystallized with ethyl acetate to obtain 1.06 g of product 3 as light yellow crystals; (3) Preparation of 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoic acid (4a) Disperse 0.50g of intermediate 3 and 0.36g of p-aminobenzoic acid in 30mL of isopropanol, add 2 drops of concentrated hydrochloric acid, heat to 85°C, reflux for 4h, cool to room temperature, filter, and wash with isopropanol and ethyl acetate Filter cake, dried to give 0.53 g of product 4a as a white solid; (4) Methyl 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoate (5a ) preparation Add 0.50g of 4a into 50mL of anhydrous methanol, then add 0.60g of thionyl chloride dropwise at 0°C, after the addition is complete, keep warm for 30min, remove the ice bath and heat to reflux, react for 5h, and depressurize the solvent After distilling off, the crude product was recrystallized with isopropanol to obtain 0.50 g of product 5a as a white solid; (5) 4-((4-((2,3-dimethyl-2H-indazol-6-yl) (methyl) amino) pyrimidin-2-yl) amino)-N-hydroxybenzamide ( 6a) Preparation KOH 28.0g and hydroxylamine hydrochloride 23.53g were dissolved in 70mL and 120mL of anhydrous methanol respectively to obtain solution A and solution B; under ice bath, solution A was added dropwise to solution B, after stirring for 2h, the precipitate was filtered off to obtain The filtrate is the methanol solution of potassium hydroxylamine; 0.20 g of compound 5a was dissolved in 30 mL of potassium hydroxylamine solution, stirred at room temperature for 1 hour, concentrated under reduced pressure to remove most of the solvent, and the remaining residue was adjusted to pH 5-6 with 1M HCl, and a white solid was precipitated , filtered, and dried the filter cake to obtain 80.6 mg of product 6a as a white solid; N-(2-aminophenyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene The preparation method of formamide (6d), the steps are as follows: (1) N-(2-aminophenyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl) Preparation of amino) benzamide (6d) Add 0.50 g of intermediate 4a to anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoro 0.49g of boric acid and 0.26g of triethylamine, activated for 30 minutes, then added 0.17g of o-phenylenediamine, reacted overnight at room temperature, after the reaction was completed, diluted with ethyl acetate, washed with water to remove N,N-dimethylformamide, The ethyl acetate layer was washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=30:1 volume ratio eluent to obtain 0.40 g of product 6d, as off-white solid; (2) (E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene base)-N-hydroxyacrylamide (10a) preparation (1) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-methoxy-N -Preparation of methylbenzamide (7a) Add 1.00 g of intermediate 4a to anhydrous N,N-dimethylformamide, under ice-cooling, add O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoro 0.98g of boric acid and 0.52g of triethylamine, activated for 30min, then added 0.30g of N,O-dimethyl hydroxylamine hydrochloride and 0.31g of triethylamine, reacted at room temperature overnight, after the reaction was completed, diluted with ethyl acetate, washed with water to remove Remove N,N-dimethylformamide, wash the ethyl acetate layer with saturated sodium bicarbonate solution and saturated NaCl solution, dry over anhydrous magnesium sulfate, remove the solvent under reduced pressure, and wash the residue with dichloromethane:methanol=35:1 volume A specific eluent was purified by column chromatography to obtain 0.78g of product 7a as a white solid; (2) Preparation of 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzaldehyde (8a) Dissolve 1.00g of compound 7a in 50mL of anhydrous tetrahydrofuran, and cool to -40°C in a low-temperature reactor; slowly add 0.26g of lithium aluminum tetrahydrogen to it in batches, and maintain this temperature for 2 hours. After the reaction, slowly and carefully The reaction was quenched with ice water until no more bubbles were generated; the reaction solution was diluted with water, extracted with ethyl acetate, the organic layers were combined, washed with saturated NaCl solution, dried over anhydrous magnesium sulfate, the solvent was concentrated under reduced pressure, and the residue was washed with dichloromethane : The eluent of methanol=40:1 volume ratio carries out column chromatography purification and obtains 0.61g product 8a, is white solid; (3) Ethyl (E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino ) phenyl) acrylate Preparation of (9a) Add 0.32 g of triethyl phosphoroacetate to 50 mL of anhydrous tetrahydrofuran, cool to -5-0°C, slowly add 0.16 g of NaH, after the addition is complete, stir for 20 min, then add 0.52 g of 8a in tetrahydrofuran dropwise, react overnight at room temperature, After completion of the reaction, add 50mL of 10% ammonium chloride solution, stir for 30min and leave to stand for stratification. After the organic phase is dried, filter and concentrate to obtain the crude product; Purification by column chromatography afforded 0.48 g of product 9a as a white solid; (4) (E)-3-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzene base)-N-hydroxyacryloyl Preparation of amine (10a) KOH 28.0g and hydroxylamine hydrochloride 23.53g were dissolved in 70mL and 120mL of anhydrous methanol respectively to obtain solution A and solution B; under ice bath, solution A was added dropwise to solution B, after stirring for 2h, the precipitate was filtered off to obtain The filtrate is the methanol solution of potassium hydroxylamine; 0.50 g of compound 9a was dissolved in 30 mL of potassium hydroxylamine solution, stirred at room temperature for 2 hours, concentrated under reduced pressure to remove most of the solvent, and the remaining residue was adjusted to pH 5-6 with 1M HCl, and a white solid was precipitated , filtered, and dried the filter cake to obtain 0.29 g of product 10a as a white solid; (three) 4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(2-(hydroxylamine )-2-oxoethyl) benzamide (13a) preparation method, the steps are as follows: (1) Methyl (4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoyl)glycinate (11a) preparation of Add 1.00 g of intermediate 4a to anhydrous N,N-dimethylformamide, under ice-cooling, add O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoro 0.98g of boric acid and 0.52g of triethylamine, activated for 30 minutes, then added 0.39g of glycine methyl ester hydrochloride, reacted overnight at room temperature, after the reaction was completed, added ethyl acetate to dilute, washed with water to remove N,N-dimethylformaldehyde Amide, ethyl acetate layer was washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, the solvent was removed under reduced pressure, and the residue was subjected to column chromatography with dichloromethane:methanol=30:1 volume ratio eluent Purification afforded 0.87 g of product 11a as a white solid; (2) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)-N-(2-(hydroxylamine )-2-oxoethyl) benzamide (13a) preparation KOH 28.0g and hydroxylamine hydrochloride 23.53g were dissolved in 70mL and 120mL of anhydrous methanol respectively to obtain solution A and solution B; under ice bath, solution A was added dropwise to solution B, after stirring for 2h, the precipitate was filtered off to obtain The filtrate is the methanol solution of potassium hydroxylamine; 0.50 g of compound 11a was dissolved in 30 mL of potassium hydroxylamine solution, stirred at room temperature for 2 hours, concentrated under reduced pressure to remove most of the solvent, and the remaining residue was adjusted to pH 5-6 with 1M HCl, and a white solid was precipitated , filtered, and the filter cake was dried to obtain 0.31 g of product 13a as a white solid; N-(2-((2-aminophenyl)amino)-2-oxoethyl)-4-((4-((2,3-dimethyl-2H-indazol-6-yl)( The preparation of methyl) amino) pyrimidin-2-yl) amino) benzamide (14a), the steps are as follows: (1) 4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)benzoyl)glycine (12a) preparation of Add 1.00g of 11a to 30mL of methanol, then add 5mL of 3M NaOH solution, stir at room temperature for 4h, after the reaction, evaporate most of the methanol under reduced pressure, and adjust the pH of the residue to 5-6 with 1M HCl solution; filter, filter The cake was washed with ice water, and after drying, 0.81 g of product 12a was obtained as a white solid, which was directly submitted to the next step; (2) N-(2-((2-aminophenyl)amino)-2-oxoethyl)-4-((4-((2,3-dimethyl-2H-indazole-6- base) (methyl) amino) pyrimidin-2-yl) amino) benzamide (14a) preparation Add 1.00 g of intermediate 12a to anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoro 0.86g of boric acid and 0.45g of triethylamine, activated for 30 minutes, then added 0.29g of o-phenylenediamine, reacted overnight at room temperature, after the reaction was completed, diluted with ethyl acetate, washed with water to remove N,N-dimethylformamide, The ethyl acetate layer was washed with saturated sodium bicarbonate solution and saturated NaCl solution, dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure, and the residue was purified by column chromatography with dichloromethane:methanol=30:1 volume ratio eluent to obtain 0.47g product 14a, pale yellow solid; (4) 7-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)- The preparation of N-hydroxyheptanamide (19f), the steps are as follows: (1) Preparation of methyl 7-(4-nitrophenoxy)heptanoate (16f) Add 1.00g of p-nitrophenol, 1.93g of methyl 7-bromoacetate and 2.98g of potassium carbonate to 50mL of acetone, raise the temperature to reflux, and react overnight. After the reaction, evaporate the solvent under reduced pressure and recrystallize with ethyl acetate 1.62 g of product 16f were obtained as a yellow solid; (2) Preparation of methyl 7-(4-aminophenoxy)heptanoate (17f) Add 1.00 g of intermediate 16f to 50 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, react overnight at room temperature, filter, wash the filter residue with methanol, combine the filtrate and washing liquid, and evaporate under reduced pressure Removal of the solvent afforded the product 0.79g17f as a light brown solid; (3) Methyl 7-(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy )-Heptanoic acid ester (18f) preparation Add 0.50g of intermediate 3, 0.52g of intermediate 17f and 0.5mL of concentrated hydrochloric acid into 50mL of isopropanol at room temperature, raise the temperature to reflux, and react overnight; : the eluent of methanol=40:1 volume ratio carries out column chromatography purification and obtains 0.61g product 18f, is a white solid; (4) 7-(4-((4-((2,3-Dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenoxy)- Preparation of N-Hydroxyheptanamide (19f) KOH 28.0g and hydroxylamine hydrochloride 23.53g were dissolved in 70mL and 120mL of anhydrous methanol respectively to obtain solution A and solution B; under ice bath, solution A was added dropwise to solution B, after stirring for 2h, the precipitate was filtered off to obtain The filtrate is the methanol solution of potassium hydroxylamine; 0.30 g of compound 18f was dissolved in 30 mL of potassium hydroxylamine solution, stirred at room temperature for 2 hours, concentrated under reduced pressure to remove most of the solvent, and the remaining residue was adjusted to pH 5-6 with 1M HCl, and a white solid was precipitated , filtered, and dried the filter cake to obtain 0.20 g of product 19f as a white solid; (5) N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)- The preparation of N ⁸ -hydroxysuberamide (23f), the steps are as follows: (1) N ⁴ -(2,3-dimethyl-2H-indazol-6-yl)-N ⁴ -methyl-N ² -(4-nitrophenyl)pyrimidine-2,4-diamine Preparation of (20) Add 1.00g of intermediate 3, 0.58g of p-nitroaniline and 0.5mL of concentrated hydrochloric acid to 70mL of isopropanol, raise the temperature to reflux, and react overnight. After the reaction, cool to room temperature, filter, and filter the cake with a small amount of isopropanol Washing, drying filter cake, obtains 1.08g product 20, is pale yellow solid; (2) N ² -(4-aminophenyl)-N ⁴ -(2,3-dimethyl-2H-indazol-6-yl)-N ⁴ -methylpyrimidine-2,4-diamine ( 21) Preparation Suspend 1.00 g of intermediate 20 in 70 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, and react overnight at room temperature. After the reaction, evaporate the solvent under reduced pressure, and weigh the residue with ethyl acetate. Crystallization afforded 0.78 g of product 21 as a light brown solid; (3) Methyl 8-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl )amino)-8-oxooctyl ester (22f) preparation Add 0.33g of monomethyl suberate to 50mL of anhydrous N,N-dimethylformamide, add O-benzotriazole-N,N,N',N'-tetramethyl After adding 0.68g of urea tetrafluoroboric acid and 0.36g of triethylamine, activate in ice bath for 30min. After activation, add 0.76g of intermediate 21, remove the ice bath, and react overnight at room temperature. After the reaction is completed, add ethyl acetate to dilute. Wash with water to remove N,N-dimethylformamide, wash the ethyl acetate layer with saturated sodium bicarbonate solution and saturated NaCl solution, dry over anhydrous magnesium sulfate, evaporate the solvent under reduced pressure, and dichloromethane:methanol=30 The eluent of 1 volume ratio carries out column chromatography purification and obtains 0.37g product 22f, is white solid; (4) N ¹ -(4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)- Preparation of N ⁸ -Hydroxysuberamide (23f) KOH 28.0g and hydroxylamine hydrochloride 23.53g were dissolved in 70mL and 120mL of anhydrous methanol respectively to obtain solution A and solution B; under ice bath, solution A was added dropwise to solution B, after stirring for 2h, the precipitate was filtered off to obtain The filtrate is the methanol solution of potassium hydroxylamine; 0.20 g of compound 22f was dissolved in 30 mL of potassium hydroxylamine solution, stirred at room temperature for 2 hours, concentrated under reduced pressure to remove most of the solvent, and the remaining residue was adjusted to pH 5-6 with 1M HCl, and a white solid was precipitated , filtered, and the filter cake was dried to give 0.14 g of product 23f as a white solid; (6) 6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino )-N-hydroxy caproamide (28e) preparation, the steps are as follows: (1) Preparation of methyl 6-((4-nitrophenyl)amino)hexyl ester (25e) Add 1.00g of p-fluoronitrobenzene, 1.54g of methyl 6-aminocaproate hydrochloride and 2.94g of potassium carbonate into 50mL of N,N-dimethylformamide, raise the temperature to 50°C overnight, and after the reaction , cooled to room temperature, the reaction solution was poured into ice water, a yellow solid was precipitated, filtered, dried, and the residue was subjected to column chromatography with an eluent of petroleum ether:ethyl acetate=4:1 volume ratio to obtain 1.32g of product 25e, It is light yellow solid; (2) Preparation of methyl 6-((4-aminophenyl)amino)hexyl ester (26e) Suspend 1.00 g of intermediate 25e in 70 mL of methanol, add 0.1 g of 10% Pd/C, pass hydrogen gas under normal pressure stirring, and react overnight at room temperature. After the reaction is completed, the solvent is evaporated under reduced pressure, and the residue is recrystallized with ethyl acetate 0.71 g of product 26e was obtained as a brown solid; (3) Methyl 6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl )amino)-hexyl ester (27e) preparation Add 0.5g of intermediate 3, 0.49g of intermediate 26e and 0.5mL of concentrated hydrochloric acid into 50mL of isopropanol at room temperature, raise the temperature to reflux, and react overnight; after the reaction, evaporate the solvent under reduced pressure, and wash the residue with dichloromethane : The eluent of methanol=40:1 volume ratio carries out column chromatography purification and obtains 0.55g product 27e, is pale yellow solid; (4) 6-((4-((4-((2,3-dimethyl-2H-indazol-6-yl)(methyl)amino)pyrimidin-2-yl)amino)phenyl)amino )-N-hydroxy caproamide (28e) preparation KOH 28.0g and hydroxylamine hydrochloride 23.53g were dissolved in 70mL and 120mL of anhydrous methanol respectively to obtain solution A and solution B; under ice bath, solution A was added dropwise to solution B, after stirring for 2h, the precipitate was filtered off to obtain The filtrate is the methanol solution of potassium hydroxylamine; 0.20 g of compound 27e was dissolved in 30 mL of potassium hydroxylamine solution, stirred at room temperature for 2 hours, concentrated under reduced pressure to remove most of the solvent, and the remaining residue was adjusted to pH 5-6 with 1M HCl, and a white solid was precipitated , filtered, and the filter cake was dried to afford 0.13 g of product 28e as a reddish solid. 6. Use of the compound as claimed in claim 1, 2 or 3 in the preparation of medicines for preventing or treating tumor-related diseases. 7. The application according to claim 6, characterized in that the tumor-related diseases are: various blood tumors or various solid tumors. 8. The application according to claim 7, characterized in that, the various types of hematological tumors are: various leukemias, myeloproliferative tumors; the various types of leukemias are: severe acute lymphoblastic leukemia, acute myeloid leukemia Leukemia, acute megakaryoblastic leukemia; the myeloproliferative tumors are: chronic myelogenous leukemia, polycythemia vera, essential thrombocythemia, myelofibrosis, chronic neutrophil leukemia, chronic eosinophilic leukemia, systemic mastocytosis and other unclassified myeloproliferative neoplasms. 9. The application according to claim 7, wherein the solid tumor is: various forms of nasopharyngeal carcinoma, renal cell carcinoma, soft tissue sarcoma, papillary thyroid carcinoma, thymoma, liver cancer, breast cancer, Melanoma, prostate cancer, or retinoblastoma. 10. A pharmaceutical composition suitable for oral or parenteral administration, comprising the compound of claim 1, 2 or 3 and one or more pharmaceutically acceptable carriers or excipients.