CN111196813B

CN111196813B - 1,3, 4-trisubstituted pyrazolopyrimidine compound as well as preparation method and application thereof

Info

Publication number: CN111196813B
Application number: CN201911126051.8A
Authority: CN
Inventors: 赵桂森; 冉凡胜; 陈鑫; 刘美霞; 李玉霞
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2018-11-20
Filing date: 2019-11-18
Publication date: 2021-10-26
Anticipated expiration: 2039-11-18
Also published as: CN109369654A; CN111196813A

Abstract

The invention relates to a1, 3, 4-trisubstituted pyrazolopyrimidine compound and a preparation method and application thereof. The compound has a structure shown in formula I. The invention also relates to a preparation method and a pharmaceutical composition of the compound containing the structure of the formula I. The invention also provides application of the compound and pharmaceutically acceptable salts thereof in preparing BTK inhibitors.

Description

1,3, 4-trisubstituted pyrazolopyrimidine compound as well as preparation method and application thereof

Technical Field

The invention relates to the field of organic compound synthesis and medical application, in particular to a1, 3, 4-trisubstituted pyrazolopyrimidine compound and a preparation method and pharmaceutical application thereof.

Background

BTK belongs to the Tec family of non-receptor tyrosine kinases, consisting of 659 amino acids, comprising 5 domains, from the amino terminus a PH domain (pleckstrin homology domain), a TH domain (Tec homology domain), a SH 3domain (Src homology 3domain), a SH 2domain (Src homology 2domain) and a catalytic domain (tyrosine kinase domain, also known as SH1 domain), respectively. (see Kawakami Y, Kitaura J, Hata D, Yao L, Kawakami T: Functions of Bruton's tyrosinase in mass and B cells. J Leukoc Biol 1999,65(3): 286) -290.) wherein the PH domain is the key domain for BTK to participate in extracellular stimulation, recognize and bind PIP 3; the TH domain comprises the BTK motif and a proline-rich conserved region; the SH3 structural domain comprises an autophosphorylation site Tyr223, can specifically recognize proline-rich fragments in the TH structural domain, and induces intramolecular folding; the SH 2domain recognizes phosphorylated tyrosine residues with a specific peptide sequence; the catalytic domain is a hinge-linked double-leaf structure, in which the N-leaf end (amino acid residue 394-474) consists of 5 beta sheets and an alpha helix; the C-lobe terminal (amino acid residue 481-659) consists of 8 alpha helices and 1 beta sheet, and contains an activation loop; the hinge domain (amino acid residue 475-480) is the ATP-binding domain. The catalytic domain and the SH 3domain respectively have a tyrosine phosphorylation site, and activated Src family kinase can phosphorylate Tyr551 of the catalytic domain, induce Tyr223 of the SH 3domain to autophosphorylate, and enable BTK to be in an activated state. (see Satterhwaite AB, Li Z, Witte ON: Btk function in B cell resolution and response. Semin Immunol 1998,10(4): 309-316). BTK is over-expressed in B cell malignant tumor cells, and the research on BTK inhibitors has important significance for treating B cell malignant tumors.

Disclosure of Invention

The invention aims to provide a1, 3, 4-trisubstituted pyrazolopyrimidine compound with BTK inhibitory activity; the invention also aims to provide a preparation method and pharmaceutical application of the 1,3, 4-trisubstituted pyrazolopyrimidine compound.

In order to achieve the purpose, the invention adopts the following technical scheme:

mono, 1,3, 4-trisubstituted pyrazolopyrimidines

The structure of the 1,3, 4-trisubstituted pyrazolopyrimidine compound or the medicinal salt thereof is shown as the general formula I:

wherein X is substituted piperidyl, substituted benzyl or substituted acetamido as shown below:

R₁is C1-6 straight chain or branched chain alkyl, hydroxyl, amino, ether substituted acyl, C1-6 straight chain or branched chain alkyl; r₂Is hydrogen, halogen, C1-6 straight chain or branched chain alkyl, cyano, trifluoromethyl or nitro; r₃Hydrogen, halogen, C1-6 straight chain or branched chain alkyl, cyano, trifluoromethyl, straight chain or branched chain alkyl substituted acyl and nitro; r₄Is hydrogen, halogen, C1-6 straight chain or branched chain alkyl; r₅Is a C1-6 straight chain or branched chain alkyl; r₆Is hydrogen, halogen, C1-6 straight chain or branched chain alkyl, trifluoromethyl, cyano, C1-6 straight chain or branched chain substituted acylamino; y is carbon and nitrogen; n is 0,1,2,3,4,5, 6.

Preferably, R₁Is acryloyl, 2-butenoyl, 2-butynoyl, 2-hydroxyacetyl, 2-hydroxypropionyl, 3-hydroxy-2, 2-dimethylpropionyl, 3-methoxypropionyl, chloroacetyl; r₂Is hydrogen, fluorine, chlorine, bromine, cyano, trifluoromethyl, nitro, methyl; r₃Is hydrogen, fluorine, bromine, cyano, acetyl, methyl, trifluoromethyl, nitro, methylamido; r₄Is hydrogen, fluorine, bromine, methyl; r₅Is methyl, ethyl, tert-butyl; r₆Is hydrogen, fluorine, chlorine, bromine, methyl, trifluoromethyl, cyano, chloroacetamide, acrylamide, 2-butenamide; y is carbon and nitrogen; n is 0,1, 2.

Further preferably, the 1,3, 4-trisubstituted pyrazolopyrimidine compound or a pharmaceutically acceptable salt thereof is selected from one of the following compounds:

TABLE 1 structural formula of object Compound

The structures of the above preferred 83 compounds and their corresponding numbers are indicated in parentheses and will be used directly in the following description for convenience of description and brevity of expression.

Preparation method of di-1, 3, 4-trisubstituted pyrazolopyrimidine compound

The preparation method of the 1,3, 4-trisubstituted pyrazolopyrimidine compound with the structure shown in the general formula I or the medicinal salt thereof comprises the following steps:

the synthetic route is as follows:

reagents and conditions: (a) phenol, sodium hydride (NaH), Tetrahydrofuran (THF),80 ℃,12 h; (b) palladium acetate (Pd (OAc)₂) 2-dicyclohexylphosphonium-2, 4, 6-triisopropylbiphenyl (X-PHOS), potassium acetate (KOAc), pinacol diboron, 1, 4-dioxane, 90 ℃,12 h; (c) n-bromosuccinimide (NBS), N, N-Dimethylformamide (DMF),80 ℃,3 h; (d) tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Potassium phosphate (K)₃PO₄)1, 4-dioxane, water 4:1v/v,135 ℃,30 h; (e) N-BOC-3-hydroxypiperidine, triphenylphosphine (PPh)₃) Diisopropyl azodicarboxylate (DIAD), THF,0 ℃ for 5 h; (f) saturated HCl 1, 4-dioxane solution, r.t.,5 h; (g) substituted carboxylic acid, HBTU, N, N-Diisopropylethylamine (DIEA), DMF, r.t.,5 h; (h) bromoacetyl bromide, sodium bicarbonate (NaHCO)₃) Ethyl acetate, water 1:1v/v, r.t.,2 h; (i) potassium carbonate (K)₂CO₃) DMF, r.t.,10 h; (j) oxalyl chloride, Dichloromethane (DCM), DMF, r.t.; (k) substituted primary amines, triethylamine (Et)₃N), THF, 0-r.t.1h; (l) Reduced iron powder, ammonium chloride, ethanol: water 3:1v/v, 90 ℃;(m) substituted acyl chloride, DIEA, THF, 0-r.t., 5h.

Wherein R is₁～R₆N is shown in the general formula I.

The method comprises the following specific steps:

(1) reaction of starting material 1 with phenol and NaH under THF conditions affords intermediate 2, intermediate 2 with Pd (OAc)₂X-PHOS, KOAc and pinacol diboron react under the condition of 1, 4-dioxane to obtain an intermediate 3; reacting the raw material 4 with NBS under the DMF condition to obtain an intermediate 5; intermediate 3 and intermediate 5 with Pd (PPh)₃)₄And K₃PO₄Reacting under the condition of 1, 4-dioxane and water to obtain an intermediate 6;

(2) reacting the intermediate 6 with N-BOC-3-hydroxypiperidine, PPh3 and DIAD under the THF condition to obtain an intermediate 7, reacting the intermediate 7 with concentrated HCl under the 1, 4-dioxane condition to obtain an intermediate 8, and condensing the intermediate 8 with various substituted carboxylic acids or substituted acyl chlorides under the alkaline condition to obtain a target compound Ra;

(3) the ethyl acetate debrominated by different substituted amines 9a-9z reacts to obtain intermediates 10a-10z, 10a-10z and an intermediate 6 at K₂CO₃Reacting under the condition to obtain a target compound Rb;

(4) the raw material 11 is reacted with oxalyl chloride to obtain acyl chloride intermediate 12, and then reacted with different substituted primary amines to obtain intermediates 13a-13c, 13a-13c and intermediates 6 and K₂CO₃Reacting under the action of DMF to obtain a target compound Rc;

(5) intermediate 5 with starting material 14 in Pd (PPh)₃)₄Obtaining an intermediate 15 for catalysis;

(6) reacting the intermediate 15 with different benzyl bromide to obtain a part of final product Rd, reducing the target compound Rd containing the nitro group to obtain a target compound Rd containing the amino group, and further reacting the target compound Rd containing the amino group with different acyl chloride to obtain a part of final product Rd;

(7) intermediate 15 and intermediates 10a-10z and K₂CO₃Reacting under the action of DMF to obtain the target compound Rg.

Preferably, the preparation method of the 1,3, 4-trisubstituted pyrazolopyrimidine compound or the pharmaceutically acceptable salt thereof comprises one of the following steps:

(1) a process for the preparation of compound Ra1-Ra9, comprising the steps of:

(i) dissolving phenol in anhydrous tetrahydrofuran, stirring in an ice bath, gradually adding sodium hydride, stirring for 30min, continuously reacting for 30min at 80 ℃, cooling to room temperature, adding a starting material 1, refluxing for 12h at 80 ℃, cooling to room temperature after the reaction is finished, quenching the reaction with water, extracting with ethyl acetate, combining organic phases, washing with saturated saline solution, drying, filtering, and evaporating the solvent under reduced pressure to obtain a colorless oily substance, performing silica gel column chromatography to obtain petroleum ether: ethyl acetate 100:1, obtaining an intermediate 2;

(ii) taking Pd (OAc)₂X-PHOS, dissolved in 1, 4-dioxane, N₂Stirring at room temperature for 20min, dissolving intermediate 2 in 1, 4-dioxane, adding pinacol diborate and KOAc, adding activated ligand solution, N₂Protecting, reacting at 90 ℃ for 12h, after the reaction is finished, carrying out hot filtration by using diatomite, carrying out reduced pressure evaporation to remove a solvent, and carrying out silica gel column chromatography on petroleum ether: ethyl acetate 100:1-40:1 to give intermediate 3;

(iii) dissolving the starting material 4 in DMF, adding NBS, stirring, heating in an oil bath at 80 ℃, changing the color of the solution from light yellow to red, reacting for 3h, detecting by TLC, reacting basically completely, cooling to room temperature, pouring the reaction solution into ice water, stirring, precipitating a large amount of yellow solid, filtering, washing a filter cake with ice water, and drying to obtain an intermediate 5;

(iv) taking intermediate 3, intermediate 5 and Pd (PPh)₃)₄、K₃PO_4.3H₂O, adding the mixture into a two-necked bottle, adding a solvent 1, 4-dioxane: dissolving 4:1(v/v) water, removing oxygen in the solution by ultrasonic treatment, and adding N₂Displacing air in the apparatus, N₂And (4) protection, heating and refluxing in an oil bath at 135 ℃, reacting for 30 hours, detecting by TLC, and basically completely reacting. Cooling the reaction solution to room temperature, filtering with diatomite, evaporating the filtrate under reduced pressure to obtain a yellow solid, and performing silica gel column chromatography on dichloromethane: methanol 100:1 to give intermediate 6;

(v) dissolving intermediate 6 in anhydrous THF, adding N-BOC-3-hydroxypiperidine, PPh₃Adding DIAD dropwise under ice bath, reacting for 5h in ice bath, evaporating the solvent under reduced pressure after the reaction is finished, and performing silica gel column chromatography on petroleum ether: ethyl acetate ═ 5:1,obtaining an intermediate 7; dissolving the intermediate 7 in a saturated HCl dioxane solution, stirring at room temperature for 5 hours, and performing suction filtration to obtain an intermediate 8 after the reaction is finished; dissolving the intermediate 8 in DMF, adding DIEA (dimethyl Ether-N-ethyl ether), substituted carboxylic acid HBTU or substituted acyl chloride, stirring at room temperature for 10h, after the reaction is finished, pouring the reaction liquid into ice water, extracting with ethyl acetate, combining organic phases, washing with water and saturated saline in sequence, drying with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure, and performing silica gel column chromatography to obtain petroleum ether: ethyl acetate is 5:1-1:1 to obtain a target product Ra;

the synthetic route is as follows:

reagents and conditions: (a) phenol, sodium hydride (NaH), Tetrahydrofuran (THF),80 ℃,12 h; (b) palladium acetate (Pd (OAc)₂) 2-dicyclohexylphosphonium-2, 4, 6-triisopropylbiphenyl (X-PHOS), potassium acetate (KOAc), pinacol diboron, 1, 4-dioxane, 90 ℃,12 h; (c) NBS, DMF,80 ℃,3 h; (d) tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Potassium phosphate (K)₃PO₄)1, 4-dioxane, water 4:1v/v,135 ℃,30 h; (e) N-BOC-3-hydroxypiperidine, triphenylphosphine (PPh)₃) Diisopropyl azodicarboxylate (DIAD), THF,0 ℃ for 5 h; (f) saturated HCl 1, 4-dioxane solution, r.t.,5 h; (g) substituted carboxylic acid, HBTU, N, N-Diisopropylethylamine (DIEA), DMF, r.t.,5 h; 0-r.t., 5 h; (m) substituted acyl chloride, DIEA, THF, 0-r.t., 5 h;

the substituted carboxylic acid is: 2-hydroxyacetic acid, 2-hydroxypropionic acid, 3-hydroxy-2, 2-dimethylpropionic acid, 3-methoxypropionic acid; the substituted acyl chlorides are: acryloyl chloride, 2-butenoyl chloride, 2-butynoyl chloride, chloroacetyl chloride;

(2) a process for the preparation of compound Rb1-Rb26, comprising the steps of:

(i) the starting material, variously substituted amines 9a-9z, was dissolved in ethyl acetate (water: 1 (v/v)) and NaHCO was added₃Stirring at room temperature, dropwise adding bromoacetyl bromide, stirring for 2h, adding water after the reaction is finished, extracting with ethyl acetate, combining organic phases, washing the organic phases with saturated saline solution, drying, filtering, and collecting filtrateEvaporating the solvent under reduced pressure to obtain different substituted bromoacetamides 10a-10 z;

(ii) dissolving intermediate 6 in DMF, adding intermediate 10a-10z, K₂CO₃Stirring at room temperature for 10h, after the reaction is finished, pouring the reaction solution into ice water, extracting with ethyl acetate, combining organic phases, washing with saturated saline solution, drying, filtering, evaporating the solvent under reduced pressure, and performing silica gel column chromatography on dichloromethane: methanol is 200:1-60:1 to obtain a target product Rb;

the synthetic route is as follows:

reagents and conditions: (h) bromoacetic acid ethyl ester, NaHCO₃Ethyl acetate, water 1:1v/v, r.t.,2 h; (i) k₂CO₃,DMF,r.t.,10h。

(3) A process for the preparation of the compound Rc1-Rc3, comprising the steps of:

(i) dissolving the starting material 11 in dichloromethane, adding oxalyl chloride and 2 drops of DMF, stirring at room temperature for reaction, detecting by TLC after 30min, and evaporating the solvent under reduced pressure to obtain an intermediate 12 after the reaction is finished. Placing substituted primary amine into a reaction bottle, adding THF (tetrahydrofuran), stirring under an ice bath condition, dropwise adding the intermediate 12 dissolved in the THF into the reaction bottle, and reacting at room temperature after dropwise adding. And (3) performing TLC detection after 1h, completely reacting, adding water into the reaction solution, extracting with ethyl acetate, combining organic phases, drying with anhydrous sodium sulfate, filtering, evaporating under reduced pressure to remove the solvent, and drying to obtain the differently substituted benzyl bromide 13a-13 c.

(ii) Intermediate 6, dissolved in DMF, was added to intermediate 13a-13c, K₂CO₃Stirring at room temperature for 10h, after the reaction is finished, pouring the reaction liquid into ice water, extracting with ethyl acetate, combining organic phases, washing with saturated saline solution, drying, filtering, evaporating the filtrate under reduced pressure to remove the solvent, and performing silica gel column chromatography on dichloromethane/methanol (150: 1-50: 1) to obtain a target product Rc;

the synthetic route is as follows:

reagents and conditions (j) oxalyl chloride, DCM, DMF, r.t.; (k) substituted primary amines, Et₃N,THF,0℃～r.t.1h；(i)K₂CO₃,DMF,r.t.,10h；

The substituted primary amine is as follows: methylamine, ethylamine, tert-butylamine;

(4) a method for preparing compound Rd1-Rd19, comprising the steps of:

(i) taking 14 parts of raw material, 5 parts of intermediate and Pd (PPh)₃)₄、K₃PO₄.3H₂Adding O, dissolving in solvent (1, 4-dioxane: water 4:1), removing oxygen with ultrasound, and adding N₂Displacing air in the apparatus, N₂And (4) protection, heating and refluxing in an oil bath at 135 ℃, reacting for 30 hours, detecting by TLC, and basically completely reacting. Cooling the reaction solution to room temperature, filtering with diatomite, evaporating the filtrate under reduced pressure, and performing silica gel column chromatography on dichloromethane: methanol 100:1, to afford intermediate 15.

(ii) Dissolving intermediate 15 in DMF, adding different substituted benzyl bromide, K₂CO₃Stirring at room temperature for 10h, after the reaction is finished, pouring the reaction liquid into ice water, extracting with ethyl acetate, combining organic phases, washing with saturated saline solution, drying, filtering, evaporating under reduced pressure to remove the solvent, performing silica gel column chromatography, and obtaining a part of target product Rd by using an elution system of dichloromethane/methanol (150: 1-60: 1).

(iii) Dissolving target compound Rd containing nitro in ethanol: adding reducing iron powder and ammonium chloride into the solution with the ratio of water to 3:1, refluxing for 5h at 90 ℃, filtering by using kieselguhr, evaporating the solvent under reduced pressure, adding water, and filtering to obtain the target compound Rd containing amino.

(iv) Dissolving an amino-containing target compound Rd in THF, adding DIEA in ice bath, dropwise adding different substituted acyl chlorides, reacting for 5 hours at room temperature, evaporating the solvent under reduced pressure, and performing column chromatography on dichloromethane: methanol 100:1-40:1 to give part of the final product Rd.

The synthetic route is as follows:

reagents and conditions: (d) tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Potassium phosphate (K)₃PO₄)1, 4-dioxane, water 4:1v/v,135 ℃,30 h; (i) potassium carbonate (K)₂CO₃) DMF, r.t.,10 h; (l) Reduced iron powder, ammonium chloride, ethanol: water 3:1v/v, 90 ℃; (m) substituted acyl chloride, DIEA, THF, 0-r.t., 5h.

The substituted acyl chloride is as follows: acryloyl chloride, 2-butenoyl chloride, chloroacetyl chloride;

(5) the preparation method of the compound Rg1-Rg26 comprises the following steps:

(i) taking 14 parts of raw material, 5 parts of intermediate and Pd (PPh)₃)₄、K₃PO₄.3H₂Adding O, adding into a two-necked bottle, adding solvent (1, 4-dioxane: water ═ 4:1v/v for dissolution, removing oxygen in the solution with ultrasound, and adding N₂Displacing air in the apparatus, N₂And (4) protection, heating and refluxing in an oil bath at 135 ℃, reacting for 30 hours, detecting by TLC, and basically completely reacting. Cooling the reaction solution to room temperature, filtering with diatomite, evaporating the filtrate under reduced pressure, and performing silica gel column chromatography on dichloromethane: methanol 100:1 to afford intermediate 15;

(ii) dissolving intermediate 15 in DMF, adding intermediate 10a-10z, K₂CO₃And stirring at room temperature for 10 hours, after the reaction is finished, pouring the reaction liquid into ice water, extracting with ethyl acetate, combining organic phases, washing with saturated saline solution, drying, filtering, evaporating the solvent under reduced pressure, and performing silica gel column chromatography to obtain dichloromethane: obtaining a target product Rg by methanol at a ratio of 200:1-60: 1;

the synthetic route is as follows:

reagents and conditions: (d) tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Potassium phosphate (K)₃PO₄)1, 4-dioxane, water 4:1v/v,135 ℃,30 h; (i) potassium carbonate (K)₂CO₃),DMF,r.t.,10h。

The room temperature of the invention is 20-30 ℃.

The following experimental examples are only for illustrating the technical effects of the present invention, but the experimental examples are not intended to limit the present invention.

Experimental example 1: test experiment of compound for BTK inhibitory activity

Experimental materials and instruments: this experiment was performed with the aid of Eurofins Pharma, british.

The experimental method comprises the following steps: BTK (h) was incubated with 8mM MPSpH7.0, 0.2mM EDTA, 250. mu. MKVEKIGEGTYGVVYK (Cdc2 peptide), 10mM magnesium acetate and [9-33P ] -ATP (specific activity and concentration as required). The reaction was initiated by adding the Mg/ATP mixture. After 40 minutes incubation at room temperature, the reaction was stopped by adding phosphoric acid to a concentration of 0.5%. 10 μ L of the reaction was then spotted onto a P30 filter pad, washed 4 times in 0.425% phosphoric acid for 4 minutes each, washed once in methanol, then dried and scintillation counted.

In the experiment, a compound test group (C), a positive control group (P) and a negative control group (N) are set. The test group is to add the compound solution to be tested (4 muL/well) with different concentrations into 384-well plate, the positive control group is to add 1 Xkinase buffer solution with the same volume, and the others are the same as the test group; the negative control group was not added with the test compound nor with the BTK kinase solution, and was replaced with 6. mu.L/well of 1 Xkinase buffer, and the rest was the same as the test group.

The formula for calculating the inhibition rate is as follows:

where c is the test group, n is the background group, and p is the blank group.

The IC was calculated by fitting a curve to the Graghpad prism6.0 software using the logarithm of the concentration as the abscissa and the inhibition as the ordinate₅₀. The experimental results of the BTK kinase inhibitory activity assay of the target compound are shown in Table 2.

TABLE 2 inhibitory Activity of the Compounds of interest on BTK

^aND：not detected

The experimental data in Table 2 show that the compounds in the general formula I all have certain inhibitory activity on BTK, and half inhibitory concentration of part of the compounds is in nanomolar level (about 30 nM). Therefore, the invention provides the application of the compound in the general formula I in preparing the medicine for resisting B cell malignant tumor.

A pharmaceutical composition for resisting B cell malignant tumor is characterized by comprising 1,3, 4-trisubstituted pyrazolopyrimidine compound shown in general formula I or pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.

Detailed Description

The present invention will be further described with reference to examples, but the following description is only for the purpose of explaining the present invention and does not limit the contents thereof. The conditions used in the examples can be further adjusted according to the existing equipment conditions, and the implementation conditions not specified are generally the conditions in routine experiments.

Example 1:

1) preparation of intermediate 2

Dissolving phenol (8.02g, 85.23mmol) in 50ml of anhydrous tetrahydrofuran, stirring in an ice bath, gradually adding sodium hydride (2.5g, 113.64mmol), stirring for 30min, continuing to react at 80 ℃ for 30min, cooling to room temperature, adding 2-fluoro-5-bromo-pyridine (10g, 56.82mmol), refluxing at 80 ℃ for 12h, after the reaction is finished, cooling to room temperature, quenching with 100ml of water, extracting with ethyl acetate (100ml × 3), combining organic phases, washing with saturated saline, drying, filtering, evaporating the solvent under reduced pressure, and performing silica gel column chromatography to obtain 12.3g of colorless oil, wherein the yield is 86%, and the elution system is petroleum ether: ethyl acetate 100: 1.

2) preparation of intermediate 3

Dissolving palladium acetate (315mg,1.4mmol) and X-PHOS (1.3g,2.8mmol) in 10ml of 1, 4-bisOxygen hexacyclic ring, N₂Stirring at room temperature for 20min, dissolving 5-bromo-2-phenoxypyridine (7g, 28mmol) in 100ml of 1, 4-dioxane, adding pinacol diboron (14.42g,56mmol) and potassium acetate (8.4g, 84mmol), and adding activated ligand solution, N₂And (3) protecting the reaction product at 90 ℃ for 12h, after the reaction is finished, performing hot filtration by using diatomite, evaporating the solvent under reduced pressure, and performing silica gel column chromatography to obtain 4.24g of a white solid, wherein the yield is 51%, and an elution system is petroleum ether: ethyl acetate 100:1-40: 1.

3) Preparation of intermediate 5

Dissolving 4-aminopyrazolo [3,4-d ] pyrimidine (5g, 37mmol) in 30ml DMF, adding NBS (7.9g, 44.4mmol), stirring, heating in an oil bath at 80 ℃, changing the color of the solution from light yellow to red, reacting for 3h, detecting by TLC, basically completely reacting, cooling to room temperature, pouring the reaction solution into 300ml ice water, stirring, precipitating a large amount of yellow solid, filtering, washing a filter cake with ice water, and drying to obtain 6.33g of light yellow solid with the yield of 79.9%.

4) Preparation of intermediate 6

Intermediate 5(2g, 9.34mmol), intermediate 3(5.55g, 18.69mmol), tetrakis (triphenylphosphine) palladium (531mg, 0.46mmol), K were collected₃PO₄.3H₂O (7.45g, 28.02mmol), in a 250ml two-necked flask, 100ml of the solvent 1, 4-dioxane: dissolving 4:1(v/v) of water, protecting nitrogen, removing oxygen in the solution by ultrasound, replacing air in a device with nitrogen, heating and refluxing by using 135 ℃ oil bath, reacting for 30h, detecting by TLC, and basically completing the reaction. Cooling the reaction liquid to room temperature, filtering with diatomite, evaporating the filtrate under reduced pressure to obtain a yellow solid, and performing silica gel column chromatography to obtain a light yellow solid 2.04g, wherein the yield is 72%, and the elution system is dichloromethane: methanol is 100: 1.

5) Preparation of intermediate 7

Dissolving intermediate 6(1.2g,3.9mmol) in 25ml of anhydrous tetrahydrofuran, adding N-BOC-3-hydroxypiperidine (2.4g, 11.7mmol) and triphenylphosphine (3.2g, 11.7mmol), dropwise adding DIAD (2.37g,11.7mmol) under ice bath, reacting for 5h, removing the solvent by evaporation under reduced pressure, and performing silica gel column chromatography to obtain a white solid 1.1g, the yield is 58%, wherein the elution system is petroleum ether: ethyl acetate ═ 5: 1.

6) preparation of intermediate 8

The intermediate 7(0.8g,1.6mmol) was dissolved in 5ml HCl saturated dioxane solution and stirred at room temperature for 5h, after the reaction was completed, 668mg of white solid was obtained by suction filtration, yield 95%.

7) Preparation of intermediates 10a-10z

Taking different substituted anilines 9a-9z (2mmol), dissolving in 4ml ethyl acetate/water 1:1(v/v), adding sodium bicarbonate (0.31g, 3.7mmol), stirring at room temperature, dropwise adding bromoacetyl bromide (0.46g, 2.3mmol), stirring for 2h, after the reaction is finished, adding 10ml water, extracting with ethyl acetate (10ml × 3), combining organic phases, washing the organic phases with saturated saline solution, drying, filtering, evaporating the filtrate under reduced pressure to remove the solvent, and obtaining an intermediate 10a-10 z.

8) Preparation of intermediate 12

The starting material 11(1g, 4.65mmol) was dissolved in 20ml dichloromethane, 2.5ml oxalyl chloride and 2 drops DMF were added, the reaction was stirred at room temperature, TLC detection was performed after 30min, and after the reaction was completed, the solvent was distilled off under reduced pressure to obtain intermediate 12.

9) Preparation of intermediates 13a-13c

The substituted primary amine (5.14mmol) was placed in a reaction flask, 9ml of THF and triethylamine (8.57mmol) were added, stirring was carried out in an ice bath, intermediate 12 dissolved in 9ml of THF was added dropwise to the reaction flask, and after completion of the dropwise addition, the reaction was carried out at room temperature. TLC detection is carried out after 1h, the reaction is completed, 30ml of water is added into the reaction liquid, ethyl acetate (30ml multiplied by 2) is used for extraction, organic phases are combined, anhydrous sodium sulfate is used for drying, filtering is carried out, the solvent is removed by evaporation under reduced pressure, and drying is carried out, so that the intermediates 13a-13c are obtained.

10) Preparation of intermediate 15

Collecting intermediate 5(2g, 9.34mmol), 4-phenoxyphenylboronic acid (4g, 18.69mmol), Pd (PPh)₃)₄(531mg，0.46mmol)、K₃PO₄.3H₂O (7.45g, 28.02mmol), in a 250ml two-necked flask, 100ml of the solvent 1, 4-dioxane: dissolving 4:1(v/v) of water, protecting nitrogen, removing oxygen in the solution by ultrasound, replacing air in a device with nitrogen, heating and refluxing by using 135 ℃ oil bath, reacting for 30h, detecting by TLC, and basically completing the reaction. Cooling the reaction solution to room temperature, filtering with diatomite, and coolingThe filtrate was evaporated under reduced pressure to give a yellow solid, which was chromatographed on silica gel to give 2.23g of a pale yellow solid, 78.8% yield, and dichloromethane as the eluting system: methanol is 100: 1.

11) Preparation of target Compound Ra

Dissolving the intermediate 8(0.42g, 1mmol) in 3ml DMF, adding substituted carboxylic acid (1.2mmol) HBTU (0.46g, 1.2mmol) or substituted acyl chloride (1.2mmol), DIEA (0.65g, 5mmol), stirring at room temperature for 10h, after the reaction is finished, pouring the reaction solution into 30ml ice water, extracting with ethyl acetate (30ml × 3), combining organic phases, washing with 20ml water and saturated saline water in sequence, drying with anhydrous sodium sulfate, filtering, evaporating the filtrate under reduced pressure to remove the solvent, and performing silica gel column chromatography to obtain the target product Ra, wherein an eluent is petroleum ether: ethyl acetate 5:1-1: 1.

Ra1 1- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one

White solid, yield 52%, mp:174-,¹H NMR(400MHz,DMSO)δ8.37(s,1H),8.27(s,1H),8.08(d,J＝5.7Hz,1H),7.46(t,J＝7.9Hz,2H),7.28–7.14(m,4H),6.87(dd,J＝16.5,10.5Hz,0.5H),6.70(dd,J＝16.4,10.8Hz,0.5H),6.10(dd,J＝28.5,16.6Hz,1H),5.72(d,J＝10.5Hz,0.5H),5.59(d,J＝10.5Hz,0.5H),4.77-4.69(m,1H),4.55(d,J＝11.4Hz,0.5H),4.18(d,J＝12.2Hz,1H),4.07(d,J＝13.5Hz,0.5H),3.72(t,J＝12Hz,0.5),3.21(q,J＝11.5Hz,1H),3.05(t,J＝10.5Hz,1H),2.30-2.21(m,1H),2.12(d,J＝11.4Hz,1H),1.94(d,J＝13.4Hz,1H),1.65-1.56(m,1H).ESI-MS:m/z 442.19(M+H⁺)。

ra2 (R) -1- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one

White solid, 42% yield, mp:160 and 165 ℃,¹H NMR(400MHz,DMSO)δ8.37(s,1H),8.27(s,1H),8.08(d,J＝5.7Hz,1H),7.46(t,J＝7.9Hz,2H),7.28-7.14(m,4H),6.87(dd,J＝16.5,10.5Hz,1H),6.10(dd,J＝28.5,16.6Hz,1H),5.72(d,J＝10.5Hz,1H),4.77-4.69(m,1H),4.55(d,J＝11.4Hz,1H),4.18(d,J＝12.2Hz,1H),3.72(t,J＝12Hz,1H),3.21(q,J＝11.5Hz,1H),2.30-2.21(m,1H),2.12(d,J＝11.4Hz,1H),1.94(d,J＝13.4Hz,1H),1.65-1.56(m,1H).ESI-MS:m/z 442.08(M+H⁺)。

ra3 (trans) 1- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) but-2-en-1-one

White solid, yield 46%, mp:198-,¹H NMR(400MHz,DMSO)δ8.36(s,1H),8.27(s,1H),8.07(d,J＝8.1Hz,1H),7.46(t,J＝7.9Hz,2H),7.26-7.16(m,4H),6.73-6.68(m,0.5H),6.57(d,J＝13.8Hz,1H),6.37(d,J＝15.6Hz,0.5H),4.75-4.65(m,1H),4.54(d,J＝12.4Hz,0.5H),4.18-4.07(m,1.5H),3.21–3.16(m,1H),3.13-3.06(m,1H),2.29-2.20(m,1H),2.13-2.09(m,1H),1.93(d,J＝12.2Hz,1H),1.86(d,J＝4.8Hz,1.5H),1.71(d,J＝4.8Hz,1.5H),1.64–1.49(m,1H).ESI-MS:m/z 456.14(M+H⁺)。

ra 41- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) but-2-yn-1-one

White solid, 56% yield, mp:120-,¹H NMR(400MHz,DMSO)δ8.38(s,1H),8.28(d,J＝11.4Hz,1H),8.08(d,J＝8.2Hz,1H),7.46(t,J＝7.5Hz,2H),7.29–7.15(m,4H),4.83-4.80(m,0.5H),4.74–4.64(m,0.5H),4.41(dd,J＝12.2,2.5Hz,0..5H),4.33–4.18(m,1H),3.96(dd,J＝8.9,4.0Hz,0.5H),3.83(dd,J＝12.9,8.9Hz,0.5H),3.35–3.25(m,1H),3.18(t,J＝10.0Hz,0.5H),2.30-2.22(m,1H),2.14(s,1H),2.06(s,1.5H),1.99(s,1H),1.83(s,1.5H),1.69-1.53(m,1H).ESI-MS:m/z 454.03(M+H⁺)。

ra5 1- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) -2-hydroxyethan-1-one

White solid, 39% yield, mp:232-,¹H NMR(400MHz,DMSO)δ8.37(s,1H),8.27(d,J＝7.0Hz,1H),8.08(dd,J＝8.1,2.3Hz,1H),7.46(t,J＝7.9Hz,2H),7.28–7.15(m,4H),4.81-4.77(m,0.5H),4.71–4.64(m,0.5H),4.48(dd,J＝11.7,2.9Hz,0.5H),4.20-3.92(m,3H),3.90(d,J＝14.0Hz,0.5H),3.72(d,J＝14.0Hz,0.5H),3.21-3.05(m,1.5H),3.01–2.78(m,1H),2.29–2.17(m,1H),2.13(s,1H),1.89(d,J＝12.4Hz,1H),1.74–1.50(m,1H).ESI-MS:m/z 446.16(M+H⁺)。

ra6 1- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) -2-hydroxypropan-1-one

White solid, yield 62%, mp:218-222 deg.C,¹H NMR(400MHz,DMSO)δ8.37(d,J＝2.0Hz,1H),8.27(d,J＝5.0Hz,1H),8.08(dd,J＝8.5,2.1Hz,1H),7.46(t,J＝7.9Hz,2H),7.21(ddd,J＝24.3,16.1,7.9Hz,4H),4.99(ddd,J＝37.6,32.5,6.7Hz,1H),4.77(s,0.5H),4.66(d,J＝10.5Hz,0.5H),4.54–4.44(m,1H),4.40–4.27(m,1H),4.19(d,J＝17.1Hz,0.5H),4.03(s,0.5H),3.23–3.08(m,1H),2.92–2.77(m,1H),2.31–2.18(m,1H),2.13(s,1H),1.92(d,J＝19.2Hz,1H),1.66(s,0.5H),1.52(s,0.5H),1.27–1.09(m,3H).ESI-MS:m/z 460.28(M+H⁺)。

ra 71- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) -3-hydroxy-2-, 2-dimethylpropan-1-one

White solid, yield 49%, mp:210- & ltSUB & gt 214 ℃,¹H NMR(400MHz,DMSO)δ(ppm):8.31(s,1H),8.19(s,1H),8.01(d,J＝8.4Hz,1H),7.38(t,J＝7.5Hz,2H),7.20–7.08(m,4H),4.64(t,J＝10.6Hz,1H),4.51(t,J＝5.6Hz,1H),4.39(d,J＝12.2Hz,1H),4.22(d,J＝12.9Hz,1H),3.36(d,J＝5.6Hz,3H),3.18(t,J＝11.6Hz,1H),2.89(t,J＝12.0Hz,1H),2.20(dd,J＝24.0,11.9Hz,1H),2.04(d,J＝9.9Hz,1H),1.83(d,J＝13.0Hz,1H),1.53(dd,J＝25.0,12.5Hz,1H),1.10(d,J＝7.0Hz,6H).¹³C NMR(100MHz,DMSO)δ(ppm):174.92(s),163.44(s),158.73(s),156.23(s),154.42(d,J＝13.3Hz),147.29(s),141.08(s),140.46(s),130.22(s),125.14(s),124.84(s),121.56(s),112.22(s),98.16(s),69.43(s),53.03(s),48.80(s),45.21(s),44.16(s),30.07(s),24.81(s),23.43(s),23.16(s).ESI-MS:m/z 487.91(M+H⁺)。

ra 81- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) -3-methoxypropan-1-one

White solid, yield 57%, mp:152-,¹H NMR(400MHz,DMSO)δ8.37(t,J＝2.3Hz,1H),8.27(d,J＝9.5Hz,1H),8.07(dt,J＝8.4,2.8Hz,1H),7.46(t,J＝7.9Hz,2H),7.26-7.16(mz,4H),4.80-4.73(m,0.5H),4.66-4.50(m,0.5H),4.51(d,J＝12.2Hz,0.5H),4.20(d,J＝13.1Hz,0.5H),4.05(dd,J＝13.0,2.5Hz,0.5H),3.92(d,J＝12.8Hz,0.5H),3.64–3.46(m,2.5H),3.22(d,J＝13.4Hz,3H),3.17–3.07(m,1H),2.87(t,J＝10.8Hz,0.5H),2.72–2.54(m,1.5H),2.42(dt,J＝15.4,6.3Hz,0.5H),2.28-2.19(m,1H),2.16–2.05(m,1H),1.94–1.82(m,1H),1.70–1.44(m,1H).ESI-MS:m/z 474.06(M+H⁺)。

ra9 1- (3- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) -2-chloroeth-1-one

White solid, 63% yield, mp:182-,¹H NMR(400MHz,DMSO)δ8.37(s,1H),8.27(d,J＝7.0Hz,1H),8.08(d,J＝5.6Hz,1H),7.46(t,J＝7.8Hz,2H),7.32–7.13(m,4H),4.90-4.83(m,0.5H),4.71–4.63(m,0.5H),4.49-4.41(m,2H),4.26(d,J＝12.9Hz,0.5H),4.17(d,J＝11.1Hz,0.5H),4.04(d,J＝13.6Hz,0.5H),3.85(d,J＝13.0Hz,0.5H),3.78–3.69(m,0.5H),3.24-3.18(m,1H),3.00–2.91(m,0.5H),2.30–2.07(m,2H),1.96–1.82(m,1H),1.78–1.69(m,0.5H),1.61–1.51(m,0.5H).ESI-MS:m/z 464.14(M+H⁺)。

12) preparation of target compound Rb

Dissolving the intermediate 6(0.30g and 1mmol) in 3ml of DMF, adding the intermediate 10a-10z (1.2mmol) and potassium carbonate (0.21g and 1.5mmol), stirring at room temperature for 10h, after the reaction is finished, pouring the reaction into 30ml of ice water, extracting with ethyl acetate (30ml multiplied by 3), combining organic phases, washing with saturated saline, drying, filtering, decompressing and distilling off the solvent from the filtrate, and carrying out silica gel column chromatography to obtain the target product Rb, wherein the elution system is dichloromethane/methanol (200: 1-60: 1).

Rb 12- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-fluorophenyl) acetamide

White solid, yield 61%, mp:198-,¹H NMR(400MHz,DMSO)δ(ppm):1H NMR(400MHz,DMSO)δ10.68(s,1H),8.40(d,J＝1.9Hz,1H),8.27(s,1H),8.10(dd,J＝8.5,2.2Hz,1H),7.56(d,J＝11.4Hz,1H),7.46(t,J＝7.8Hz,2H),7.38(dd,J＝15.0,7.9Hz,2H),7.31(d,J＝7.5Hz,1H),7.22(td,J＝14.9,8.0Hz,5H),6.92(t,J＝8.3Hz,1H),5.26(s,2H).¹³C NMR(100MHz,DMSO)δ(ppm):165.95(s),163.46(s),158.73(s),156.51(s),155.69(s),154.32(s),147.17(s),141.51(s),140.37(s),131.04(d,J＝9.7Hz),130.23(s),125.10(s),124.70(d,J＝2.9Hz),121.58(s),115.43(s),112.26(s),110.69(s),106.59(s),97.99(s),50.21(s).ESI-MS:m/z 456.19(M+H⁺)。

rb 22- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-fluorophenyl) acetamide

White solid, yield 43%, mp:110-,¹H NMR(400MHz,DMSO)δ(ppm):10.68(s,1H),8.40(d,J＝2.4Hz,1H),8.27(s,1H),8.10(dd,J＝8.5,2.4Hz,1H),7.56(d,J＝11.6Hz,1H),7.46(t,J＝7.9Hz,2H),7.38(dd,J＝15.0,8.0Hz,1H),7.31(d,J＝8.4Hz,1H),7.26-7.17(m,4H),6.92(td,J＝8.3,1.9Hz,1H),5.27(s,2H).¹³C NMR(100MHz,DMSO)δ(ppm):165.94(s),163.46(s),162.58(d,J＝240Hz),158.73(s),156.51(s),155.69(s),154.32(s),147.17(s),141.51(s),140.72(d,J＝11Hz),140.36(s),131.03(d,J＝9Hz),130.22(s),125.09(s),124.68(s),121.58(s),115.41(s),112.25(s),110.58(d,J＝21Hz),106.46(d,J＝26Hz),98.00(s),50.20(s).ESI-MS:m/z 456.11(M+H⁺)。

rb 32- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-fluorobenzyl) acetamide

White solid, yield 70%, mp:232-,¹H NMR(400MHz,DMSO)δ(ppm):8.75(t,J＝5.9Hz,1H),8.39(d,J＝2.2Hz,1H),8.27(s,1H),8.08(dd,J＝8.5,2.4Hz,1H),7.46(t,J＝7.9Hz,2H),7.37(q,J＝8Hz,1H),7.27-7.13(m,6H),7.08(t,J＝8.3Hz,1H),5.10(s,2H),4.34(d,J＝5.9Hz,2H).¹³C NMR(100MHz,DMSO)δ(ppm):167.09(s),162.73(d,J＝241Hz),163.43(s),158.71(s),156.40(s),155.54(s),154.34(s),147.17(s),142.55(d,J＝7Hz),141.46(s),140.34(s),130.66(d,J＝8Hz),130.22(s),125.09(s),124.74(s),123.56(d,J＝3Hz),121.56(s),114.13(d,J＝43Hz),114.12(s),112.24(s),98.09(s),49.77(s),42.16(s).ESI-MS:m/z 470.20(M+H⁺)。

rb 42- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-fluorophenethyl) acetamide

White solid, yield 61%, mp:192-,¹H NMR(400MHz,DMSO)δ(ppm):8.38(d,J＝2.1Hz,1H),8.27(s,1H),8.23(t,J＝4.5Hz,1H),8.07(dd,J＝8.5,2.3Hz,1H),7.46(t,J＝7.8Hz,2H),7.27-7.17(m,6H),7.09(t,J＝8.8Hz,2H),4.97(s,2H),3.30(q,J＝6.8Hz,2H),2.72(t,J＝7.1Hz,2H).¹³C NMR(100MHz,DMSO)δ(ppm):166.69(s),163.43(s),161.46(d,J＝240Hz),158.68(s),156.40(s),155.53(s),154.33(s),147.17(s),141.37(s),140.35(s),135.88(d,J＝2.9Hz),130.94(d,J＝7.9Hz),130.22(s),125.09(s),124.76(s),121.57(s),115.52(s),115.31(s),112.23(s),98.01(s),49.55(s),34.48(s).ESI-MS:m/z 484.30(M+H⁺)。

rb 52- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chlorophenyl) acetamide

White solid, yield 67%, mp:232-,¹H NMR(400MHz,DMSO)δ(ppm):10.59(s,1H),8.40(d,J＝2.0Hz,1H),8.27(s,1H),8.09(dd,J＝8.5,2.2Hz,1H),7.61(d,J＝8.8Hz,2H),7.46(t,J＝7.8Hz,2H),7.39(d,J＝8.8Hz,2H),7.26-7.17(m,4H),5.25(s,2H).¹³C NMR(100MHz,DMSO)δ(ppm):165.71(s),163.46(s),158.72(s),156.50(s),155.68(s),154.32(s),147.17(s),141.49(s),140.36(s),137.99(s),130.22(s),129.25(s),127.64(s),125.10(s),124.68(s),121.58(s),121.21(s),112.25(s),98.00(s),50.19(s).ESI-MS:m/z 472.08(M+H⁺)。

rb 62- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chloro-4-fluorophenyl) acetamide

White solid, 56% yield, mp:212-,¹H NMR(400MHz,DMSO)δ(ppm):1H NMR(400MHz,DMSO)δ10.68(s,1H),8.40(d,J＝2.3Hz,1H),8.27(s,1H),8.09(dd,J＝8.5,2.4Hz,1H),7.89(dd,J＝6.8,2.4Hz,1H),7.48-7.38(m,4H),7.26-7.17(m,4H),5.26(s,2H).¹³C NMR(100MHz,DMSO)δ(ppm):165.89(s),163.47(s),158.72(s),156.51(s),155.71(s),154.31(s),152.53(s),147.17(s),141.55(s),140.36(s),136.26(s),130.22(s),125.10(s),124.66(s),121.80(s),121.58(s),121.11(s),120.03(s),119.61(s),117.69(s),112.25(s),98.00(s),50.13(s).ESI-MS:m/z 490.06(M+H⁺)。

rb7 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-cyanophenyl) acetamide

White solid, yield 68%, mp:224-,¹H NMR(400MHz,DMSO)δ(ppm):1H NMR(400MHz,DMSO)δ10.91(s,1H),8.40(d,J＝2.1Hz,1H),8.27(s,1H),8.10(dd,J＝8.5,2.4Hz,1H),7.78(q,J＝8.9Hz,4H),7.46(t,J＝7.9Hz,2H),7.22(td,J＝15.0,8.0Hz,5H),5.31(s,2H).¹³C NMR(100MHz,DMSO)δ166.43(s),163.48(s),158.73(s),156.53(s),155.68(s),154.31(s),147.17(s),143.18(s),141.57(s),140.36(s),133.90(s),130.22(s),125.10(s),124.64(s),121.59(s),119.73(s),119.41(s),112.26(s),105.89(s),98.00(s),50.31(s).ESI-MS:m/z 463.19(M+H⁺)。

rb 82- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chloro-4-cyanophenyl) acetamide

White solid, yield 54%, mp:152-,¹H NMR(400MHz,DMSO)δ(ppm):11.10(s,1H),8.40(d,J＝2.2Hz,1H),8.27(s,1H),8.09(dd,J＝8.5,2.4Hz,1H),8.02(d,J＝1.7Hz,1H),7.93(d,J＝8.6Hz,1H),7.61(dd,J＝8.6,1.8Hz,1H),7.46(t,J＝7.9Hz,2H),7.26–7.17(m,4H),5.32(s,2H).¹³C NMR(100MHz,DMSO)δ166.87(s),163.49(s),158.73(s),156.56(s),155.73(s),154.30(s),147.17(s),144.23(s),141.67(s),140.36(s),136.58(s),135.83(s),130.22(s),125.11(s),124.61(s),121.59(s),119.67(s),118.35(s),116.62(s),112.26(s),106.37(s),97.99(s),50.34(s).ESI-MS:m/z 497.15(M+H⁺)。

rb 92- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3, 4-dicyanophenyl) acetamide

White solid, yield 35%, mp:184- & 189 ℃,¹H NMR(400MHz,DMSO)δ(ppm):11.25(s,1H),8.40(d,J＝2.0Hz,1H),8.27(s,1H),8.25(d,J＝1.5Hz,1H),8.09(d,J＝8.7Hz,2H),7.96(dd,J＝8.7,1.7Hz,1H),7.46(t,J＝7.8Hz,2H),7.28–7.16(m,4H),5.34(s,2H).¹³C NMR(100MHz,DMSO)δ(ppm):167.05(s),163.50(s),158.73(s),156.58(s),155.74(s),154.29(s),147.16(s),143.42(s),141.72(s),140.35(s),135.68(s),130.23(s),125.11(s),124.59(s),123.64(s),123.54(s),121.59(s),116.53(s),116.53(s),116.24(s),112.26(s),108.78(s),98.00(s),50.33(s).ESI-MS:m/z 488.19(M+H⁺)。

rb10 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-cyano-3- (trifluoromethyl) phenyl) acetamide

White solid, yield 59%, mp:130-,¹H NMR(400MHz,DMSO)δ(ppm):11.27(s,1H),8.40(d,J＝2.2Hz,1H),8.27(s,2H),8.13(d,J＝8.6Hz,1H),8.09(dd,J＝8.5,2.4Hz,1H),7.95(dd,J＝8.6,1.6Hz,1H),7.46(t,J＝7.9Hz,2H),7.26–7.17(m,4H),5.34(s,2H).¹³C NMR(100MHz,DMSO)δ(ppm):167.09(s),163.50(s),158.73(s),156.58(s),155.74(s),154.30(s),147.17(s),143.52(s),141.71(s),140.36(s),137.14(s),132.45(s),130.22(s),125.11(s),124.60(s),124.19(s),122.63(s),121.59(s),116.97(s),116.15(s),112.26(s),102.55(s),98.00(s),50.34(s).ESI-MS:m/z 531.09(M+H⁺)。

rb11 4- (2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) acetamido) -2-fluoro-N-methylbenzamide

White solid, yield 60%, mp:206-,¹H NMR(400MHz,DMSO)δ(ppm):10.86(s,1H),8.41(d,J＝7.9Hz,1H),8.28(d,J＝10.6Hz,1H),8.11(s,2H),7.72–7.59(m,2H),7.52–7.42(m,2H),7.35(t,J＝8.3Hz,1H),7.24-7.17(m,5H),5.29(s,2H),2.77(s,3H).¹³C NMR(100MHz,DMSO)δ(ppm):166.31(s),163.90(s),163.49(s),161.10(s),158.74(s),156.54(s),155.72(s),154.32(s),147.18(s),142.34(s),141.58(s),140.38(s),131.45(s),130.23(s),125.11(s),124.66(s),121.60(s),118.77(s),115.12(s),112.27(s),106.70(s),98.01(s),50.25(s),26.77(s).ESI-MS:m/z 513.04(M+H⁺)。

rb12 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3- (trifluoromethyl) phenyl) acetamide

White solid, yield 64%, mp:188-,¹H NMR(400MHz,DMSO)δ10.81(s,1H),8.39(d,J＝2.0Hz,1H),8.27(s,1H),8.13–8.04(m,2H),7.75(d,J＝8.1Hz,1H),7.58(t,J＝7.9Hz,1H),7.46(t,J＝7.8Hz,3H),7.26-7.17(m,4H),5.28(s,2H).¹³C NMR(100MHz,DMSO)δ166.25(s),163.48(s),158.74(s),156.53(s),155.74(s),154.33(s),147.18(s),141.57(s),140.37(s),139.78(s),130.65(s),130.23(s),129.85(s),125.85(s),125.11(s),124.68(s),123.22(s),121.59(s),120.52-120.46(m),115.76-115.66(m),112.26(s),98.01(s),50.20(s).ESI-MS:m/z 506.13(M+H⁺)。

rb13 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (2-bromophenyl) acetamide

White solid, yield 63%, mp:104-,¹H NMR(400MHz,DMSO)δ9.84(s,1H),8.40(d,J＝1.8Hz,1H),8.29(s,1H),8.10(dd,J＝8.5,2.2Hz,1H),7.67(d,J＝8.0Hz,2H),7.46(t,J＝7.8Hz,2H),7.37(t,J＝7.5Hz,1H),7.27–7.11(m,6H),5.32(s,2H).¹³C NMR(100MHz,DMSO)δ166.07(s),163.49(s),158.75(s),156.57(s),155.73(s),154.34(s),147.21(s),141.70(s),140.39(s),136.11(s),133.24(s),130.23(s),128.59(s),127.65(s),126.93(s),126.93(s),125.11(s),124.69(s),121.59(s),117.67(s),112.27(s),98.05(s),49.92(s).ESI-MS:m/z 516.09(M+H⁺)。

rb14 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-bromo-2-methylphenyl) acetamide

White solid, yield 61%, mp:274-,¹H NMR(400MHz,DMSO)δ9.99(s,1H),8.40(s,1H),8.29(s,1H),8.10(d,J＝8.2Hz,1H),7.46(s,3H),7.38(d,J＝7.5Hz,1H),7.30–7.08(m,6H),5.29(s,2H),2.31(s,3H).¹³C NMR(100MHz,DMSO)δ166.02(s),163.48(s),158.74(s),156.50(s),155.68(s),154.33(s),147.19(s),141.55(s),140.37(s),137.55(s),132.71(s),130.23(s),130.15(s),127.85(s),125.58(s),125.25(s),125.11(s),124.73(s),121.60(s),112.27(s),98.09(s),49.92(s),18.67(s).ESI-MS:530.09m/z(M+H⁺)。

rb15 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chloro-2-fluorophenyl) acetamide

White solid, yield 57%, mp:214- & 218 ℃,¹H NMR(400MHz,DMSO)δ10.43(d,J＝10.1Hz,1H),8.39(d,J＝9.6Hz,1H),8.27(d,J＝10.6Hz,1H),8.09(t,J＝8.3Hz,1H),7.85(d,J＝7.2Hz,1H),7.49–7.41(m,2H),7.40–7.28(m,2H),7.19(dd,J＝19.5,12.2Hz,6H),5.35(d,J＝10.4Hz,2H).¹³C NMR(100MHz,DMSO)δ166.35(s),163.47(s),158.74(s),156.53(s),155.69(s),154.30(s),150.88(s),147.17(s),141.57(s),140.37(s),130.22(s),127.62(s),126.24(s),125.54(s),125.10(s),124.66(s),123.01(s),121.59(s),120.30(s),112.25(s),97.99(s),49.95(s).ESI-MS:m/z 490.18(M+H⁺)。

rb16 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-bromo-3- (trifluoromethyl) phenyl) acetamide

White solid, yield 63%, mp:116-,¹H NMR(400MHz,DMSO)δ10.93(d,J＝8.3Hz,1H),8.41(d,J＝7.2Hz,1H),8.29(d,J＝9.1Hz,1H),8.20(d,J＝7.2Hz,1H),8.11(t,J＝7.5Hz,1H),7.86(t,J＝8.5Hz,1H),7.74(d,J＝6.2Hz,1H),7.47(d,J＝7.5Hz,2H),7.21(dd,J＝21.9,8.1Hz,4H),5.30(d,J＝8.3Hz,2H).¹³C NMR(100MHz,DMSO)δ166.38(s),163.49(s),158.74(s),156.55(s),155.74(s),154.32(s),147.18(s),141.63(s),140.36(s),138.91(s),136.13(s),130.25(s),130.22(s),130.25–127.91(m),125.10(s),124.61(d,J＝10.5Hz),121.58(s),118.60(s),112.34(d,J＝15.8Hz),98.02(s),50.24(s).ESI-MS:m/z 584.00(M+H⁺)。

rb17 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-nitrophenyl) acetamide

White solid, yield 78%, mp:215-,¹H NMR(400MHz,DMSO)δ10.98(s,1H),8.61(s,1H),8.40(s,1H),8.27(s,1H),8.10(dd,J＝8.5,2.2Hz,1H),7.93(dd,J＝14.5,8.2Hz,2H),7.63(t,J＝8.1Hz,1H),7.46(t,J＝7.4Hz,2H),7.26-7.17(m,5H),5.31(s,2H).¹³C NMR(100MHz,DMSO)δ166.42(s),163.50(s),158.75(s),156.55(s),155.76(s),154.33(s),148.45(s),147.19(s),141.63(s),140.39(s),140.11(s),130.85(s),130.23(s),125.66(s),125.11(s),124.67(s),121.59(s),118.67(s),113.81(s),112.27(s),98.03(s),50.24(s).ESI-MS:m/z 483.15(M+H⁺)。

rb18 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (p-tolyl) acetamide

White solid, yield 63%, mp:238-,¹H NMR(400MHz,DMSO)δ10.33(s,1H),8.39(s,1H),8.26(s,1H),8.09(d,J＝8.5Hz,1H),7.45(t,J＝7.1Hz,4H),7.28–7.15(m,5H),7.12(d,J＝8.1Hz,2H),5.22(s,2H),2.25(s,3H).¹³C NMR(100MHz,DMSO)δ165.25(s),163.46(s),158.73(s),156.47(s),155.69(s),154.35(s),147.18(s),141.41(s),140.37(s),136.57(s),133.01(s),130.23(s),129.69(s),125.09(s),124.74(s),121.58(s),119.65(s),112.26(s),98.01(s),50.16(s),20.91(s).ESI-MS:m/z 452.30(M+H⁺)。

rb19 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-cyanophenyl) acetamide

White solid, yield 55%, mp:218-,¹H NMR(400MHz,DMSO)δ10.81(s,1H),8.39(s,1H),8.27(s,1H),8.12–8.02(m,2H),7.80(d,J＝3.1Hz,1H),7.55(s,2H),7.46(t,J＝7.7Hz,2H),7.26-7.17(m,4H),5.28(s,2H).¹³C NMR(100MHz,DMSO)δ166.29(s),163.49(s),158.74(s),156.54(s),155.75(s),154.34(s),147.18(s),141.59(s),140.37(s),139.80(s),130.89(s),130.23(s),127.70(s),125.10(s),124.68(s),124.28(s),122.37(s),121.59(s),119.05(s),112.27(s),112.17(s),98.02(s),50.20(s).ESI-MS:m/z 463.25(M+H⁺)。

rb20 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (2,3, 4-trifluorophenyl) acetamide

White solid, yield 59%, mp:234-,¹H NMR(400MHz,DMSO)δ10.45(s,1H),8.39(d,J＝1.9Hz,1H),8.27(s,1H),8.09(dd,J＝8.5,2.2Hz,1H),7.64(d,J＝7.5Hz,1H),7.46(t,J＝7.8Hz,2H),7.34–7.27(m,1H),7.21(td,J＝14.7,8.0Hz,5H),5.33(s,2H).¹³C NMR(100MHz,DMSO)δ166.37(s),163.49(s),158.74(s),156.53(s),155.72(s),154.34(s),147.19(s),141.60(s),140.37(s),130.23(s),125.10(s),124.68(s),124.03(s),121.58(s),119.01(s),112.44(s),112.27(s),98.02(s),49.83(s).ESI-MS:m/z 492.04(M+H⁺)。

rb21 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4- (trifluoromethyl) phenyl) acetamide

White solid, yield 67%, mp:236-,¹H NMR(400MHz,DMSO)δ10.82(s,1H),8.39(d,J＝1.9Hz,1H),8.25(d,J＝14.5Hz,1H),8.09(dd,J＝8.5,2.2Hz,1H),7.79(d,J＝8.5Hz,2H),7.70(d,J＝8.6Hz,2H),7.46(t,J＝7.8Hz,2H),7.21(td,J＝15.0,7.9Hz,4H),5.29(s,2H).¹³C NMR(100MHz,DMSO)δ166.25(s),163.49(s),158.75(s),156.53(s),155.72(s),154.34(s),147.19(s),142.60(s),141.56(s),140.37(s),130.23(s),126.69(d,J＝3.6Hz),125.10(s),124.69(s),121.58(s),119.62(s),112.27(s),98.02(s),50.27(s).ESI-MS:m/z 506.23(M+H⁺)。

rb22 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (2-fluoro-5- (trifluoromethyl) phenyl) acetamide

White solid, yield 66%, mp:228-,¹H NMR(400MHz,DMSO)δ10.95(s,1H),8.39(d,J＝1.6Hz,1H),8.27(s,1H),8.19(s,1H),8.09(dd,J＝8.5,1.9Hz,1H),7.81(d,J＝8.6Hz,1H),7.69(d,J＝8.8Hz,1H),7.46(t,J＝7.7Hz,2H),7.21(td,J＝14.9,8.0Hz,5H),5.29(s,2H).¹³C NMR(100MHz,DMSO)δ166.37(s),163.49(s),158.74(s),156.55(s),155.75(s),154.34(s),147.19(s),141.62(s),140.37(s),138.49(s),132.77(s),130.23(s),127.44(s),125.10(s),124.61(dd,J＝24.4,8.9Hz),121.58(s),118.28(s),112.27(s),98.02(s),50.21(s).ESI-MS:m/z 524.15(M+H⁺)。

rb23 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-nitrophenyl) acetamide

White solid, yield 50%, mp:140-,¹H NMR(400MHz,DMSO)δ11.09(s,1H),8.40(d,J＝2.1Hz,1H),8.31–8.18(m,3H),8.10(dd,J＝8.5,2.3Hz,1H),7.83(d,J＝9.1Hz,2H),7.46(t,J＝7.8Hz,2H),7.26-7.17(m,4H),5.34(s,2H).¹³C NMR(100MHz,DMSO)δ166.62(s),163.50(s),158.69(s),156.48(s),155.71(s),154.33(s),147.19(s),145.12(s),142.98(s),141.65(s),140.37(s),130.23(s),125.56(s),125.11(s),124.64(s),121.59(s),119.48(s),112.27(s),98.02(s),50.37(s).ESI-MS:m/z 483.15(M+H⁺)。

rb24 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3, 5-dimethylphenyl) acetamide

White solid, yield 47%, mp:204-,¹H NMR(400MHz,DMSO)δ10.26(s,1H),8.32(s,1H),8.19(s,1H),8.02(dd,J＝8.5,2.2Hz,1H),7.39(t,J＝7.8Hz,2H),7.20–7.09(m,6H),6.64(s,1H),5.15(s,2H),2.15(s,6H).¹³C NMR(101MHz,DMSO)δ165.38(s),163.46(s),158.72(s),156.46(s),155.74(s),154.35(s),147.17(s),141.42(s),140.37(s),138.92(s),138.27(s),130.23(s),125.57(s),125.10(s),124.75(s),121.57(s),117.43(s),112.27(s),98.00(s),50.20(s),21.54(s).ESI-MS:m/z 466.25(M+H⁺)。

rb25 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-bromophenyl) acetamide

White solid, yield 44%, mp:242-245 ℃,¹H NMR(400MHz,DMSO)δ10.52(s,1H),8.32(d,J＝2.2Hz,1H),8.19(s,1H),8.02(dd,J＝8.5,2.4Hz,1H),7.51–7.41(m,5H),7.38(t,J＝7.9Hz,2H),7.14(td,J＝15.1,7.9Hz,4H),5.18(s,2H).¹³C NMR(100MHz,DMSO)δ165.74(s),163.48(s),158.73(s),156.51(s),155.70(s),154.34(s),147.18(s),141.51(s),140.37(s),138.41(s),132.16(s),130.23(s),125.10(s),124.70(s),121.62(s),121.58(s),115.70(s),112.27(s),98.02(s),50.21(s).ESI-MS:m/z 516.09(M+H⁺)。

rb26 2- (4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-bromophenyl) acetamide

White solid, yield 67%, mp:163-,¹H NMR(400MHz,DMSO)δ10.64(s,1H),8.39(d,J＝2.1Hz,1H),8.26(s,1H),8.09(dd,J＝8.5,2.4Hz,1H),7.93(s,1H),7.47(dd,J＝14.7,7.1Hz,3H),7.34–7.15(m,7H),5.25(s,2H).¹³C NMR(100MHz,DMSO)δ165.97(s),163.49(s),158.74(s),156.52(s),155.72(d,J＝4.1Hz),154.34(s),147.18(s),141.55(s),140.78–140.65(m),140.48(d,J＝21.9Hz),131.37(s),130.23(s),126.73(s),125.10(s),124.70(s),122.06(d,J＝4.9Hz),121.58(s),118.45(s),112.27(s),98.02(s),50.21(s).ESI-MS:m/z 516.00(M+H⁺)。

13) preparation of the target Compound Rc

Intermediate 6(0.30g, 1mmol) was dissolved in 3ml DMF and intermediates 13a-13c (1.2mmol), K were added₂CO₃(0.21g, 1.5mmol) at room temperatureStirring for 10h, after the reaction is finished, pouring the reaction into 30ml of ice water, extracting with ethyl acetate (30ml × 3), combining organic phases, washing with saturated saline solution, drying, filtering, evaporating the filtrate under reduced pressure to remove the solvent, and carrying out silica gel column chromatography to obtain a target product Rc, wherein an elution system is dichloromethane/methanol (150: 1-50: 1).

Rc1- ((4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -N-methylbenzamide

White solid, yield 53%, mp:116-,¹H NMR(400MHz,DMSO)δ(ppm):8.43–8.37(m,2H),8.30(s,1H),8.08(dd,J＝8.5,2.4Hz,1H),7.77(d,J＝8.1Hz,2H),7.45(t,J＝7.8Hz,2H),7.35(d,J＝8.1Hz,2H),7.27-7.14(m,4H),5.62(s,2H),2.76(d,J＝4.5Hz,3H).¹³C NMR(100MHz,DMSO)δ(ppm):166.73(s),163.46(s),158.75(s),156.64(s),154.97(s),154.32(s),147.26(s),141.58(s),140.43(s),140.39(s),134.35(s),130.20(s),127.93(s),127.82(s),125.08(s),124.68(s),121.56(s),112.22(s),98.06(s),50.00(s),26.68(s).ESI-MS:m/z 452.14(M+H⁺)。

rc 2- ((4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -N-ethylbenzamide

White solid, yield 49%, mp:171-,¹H NMR(400MHz,DMSO)δ(ppm):8.42(t,J＝5.4Hz,1H),8.38(d,J＝2.2Hz,1H),8.30(s,1H),8.08(dd,J＝8.5,2.4Hz,1H),7.78(d,J＝8.2Hz,2H),7.45(t,J＝7.9Hz,2H),7.35(d,J＝8.1Hz,2H),7.27–7.14(m,4H),5.62(s,2H),3.31–3.21(m,2H),1.10(t,J＝7.2Hz,3H).¹³C NMR(100MHz,DMSO)δ(ppm):166.04(s),163.45(s),158.74(s),156.62(s),154.97(s),154.32(s),147.26(s),141.57(s),140.43(s),140.35(s),134.53(s),130.20(s),127.90(s),127.88(s),125.07(s),124.68(s),121.56(s),112.21(s),98.05(s),50.02(s),34.46(s),15.25(s).ESI-MS:m/z 466.12(M+H⁺)。

rc 3- ((4-amino-3- (6-phenoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -N- (tert-butyl) benzamide

White solid, yield 58%, mp:182-,¹H NMR(400MHz,DMSO)δ(ppm):8.37(d,J＝2.2Hz,1H),8.29(s,1H),8.07(dd,J＝8.5,2.4Hz,1H),7.72(s,1H),7.70(d,J＝3.4Hz,2H),7.45(t,J＝7.9Hz,2H),7.32(d,J＝8.2Hz,2H),7.26–7.14(m,5H),5.61(s,2H),1.34(s,9H).¹³C NMR(100MHz,DMSO)δ(ppm):166.54(s),163.46(s),158.75(s),156.62(s),154.97(s),154.34(s),147.25(s),141.54(s),140.42(s),140.06(s),135.80(s),130.22(s),128.11(s),127.75(s),125.09(s),124.70(s),121.57(s),112.23(s),98.06(s),51.19(s),50.07(s),29.02(s).ESI-MS:m/z 493.81(M+H⁺)。

14) preparation of Compound of interest Rd

Intermediate 15(0.30g, 1mmol) was dissolved in 3ml DMF and the variously substituted benzyl bromide (1.2mmol), K was added₂CO₃(0.21g, 1.5mmol), stirring at room temperature for 10h, after the reaction is finished, pouring the reaction liquid into 30ml ice water, extracting with ethyl acetate (30ml × 3), combining organic phases, washing with saturated saline, drying, filtering, evaporating the solvent under reduced pressure, and performing silica gel column chromatography, wherein an elution system is dichloromethane/methanol (150: 1-60: 1), so as to obtain a target product Rd; dissolving a target compound Rd (1mmol) containing a nitro group in ethanol: adding reducing iron powder (2mmol) and ammonium chloride (3mmol) into the solution with the ratio of water to the solution being 3:1, refluxing for 5h at 90 ℃, filtering by using kieselguhr, evaporating the solvent under reduced pressure, adding water, and filtering to obtain an amino-containing target compound Rd; dissolving an amino-containing target compound Rd (0.5mmol) in THF, adding DIEA (1mmol) in ice bath, dropwise adding different substituted acyl chlorides (0.6mmol), reacting at room temperature for 5h, evaporating the solvent under reduced pressure, and carrying out column chromatography on dichloromethane: methanol 100:1-40:1 to give part of the final product Rd.

Rd1 1- (3-aminobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 52%, mp:160 and 162 deg.C,¹H NMR(400MHz,DMSO)δ8.28(s,1H),7.66(d,J＝8.7Hz,2H),7.47–7.40(m,2H),7.20-7.11(m,5H),6.94(t,J＝8.0Hz,1H),6.46-6.43(m,3H),5.37(s,2H),5.11(s,2H).¹³C NMR(100MHz,DMSO)δ158.66,157.55,156.79,156.38,154.75,149.31,143.79,138.20,130.61,130.53,129.46,128.40,124.26,119.46,119.44,115.48,113.57,113.24,97.69,50.51.ESI-MS:m/z 409.11(M+H⁺)。

rd 2N- (3- ((4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) phenyl) acrylamide

White solid, yield 53%, mp:173-,¹H NMR(400MHz,DMSO)δ10.14(s,1H),8.30(s,1H),7.67(d,J＝7.6Hz,3H),7.52(s,1H),7.43(t,J＝7.4Hz,2H),7.29(t,J＝7.7Hz,1H),7.24–7.08(m,5H),7.03(d,J＝7.3Hz,1H),6.39(dd,J＝16.9,10.1Hz,1H),6.23(d,J＝16.8Hz,1H),5.73(d,J＝10.1Hz,1H),5.53(s,2H).¹³C NMR(100MHz,DMSO)δ163.60,158.69,157.61,156.76,156.51,154.85,144.10,139.73,138.28,132.24,130.61,129.52,128.30,127.45,124.27,123.34,119.46,119.00,118.75,97.72,50.24.ESI-MS:m/z 463.04(M+H⁺)。

rd3 (trans) N- (4- ((4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) phenyl) -2-butenamide

White solid, yield 56%, mp:127-,¹H NMR(400MHz,DMSO)δ9.95(s,1H),8.29(s,1H),7.67(d,J＝8.4Hz,2H),7.63(d,J＝8.2Hz,1H),7.51(s,1H),7.43(t,J＝7.8Hz,2H),7.26(t,J＝7.9Hz,1H),7.18-7.11(m,5H),7.00(d,J＝7.6Hz,1H),6.77(dq,J＝13.9,6.8Hz,1H),6.08(d,J＝15.3Hz,1H),5.52(s,2H),1.84(d,J＝6.6Hz,3H).¹³C NMR(100MHz,DMSO)δ163.93(s),158.69(s),157.60(s),156.76(s),156.49(s),154.84(s),144.08(s),140.42(s),140.01(s),138.20(s),132.91(s),130.61,130.55,129.43(s),128.31(s),126.39(s),124.27(s),122.99(s),119.46(s),118.90(s),118.66,97.72(s),50.26(s),17.99(s).ESI-MS:m/z 477.12(M+H⁺)。

rd 4N- (3- ((4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) phenyl) -2-chloroacetamide

White solid, yield 60%, mp:166-,¹H NMR(400MHz,DMSO)δ10.30(s,1H),8.29(s,1H),7.67(d,J＝8.6Hz,2H),7.56(d,J＝8.8Hz,1H),7.43(t,J＝7.9Hz,3H),7.29(t,J＝7.9Hz,1H),7.20-7.11(m,5H),7.05(d,J＝7.7Hz,1H),5.53(s,2H),4.20(s,2H).¹³C NMR(100MHz,CDCl₃)δ163.76,158.80,156.87,156.21,154.19,144.81,137.36,137.01,130.01,129.96,129.63,127.16,124.90,124.16,119.77,119.62,119.14,98.26,50.70,42.85.ESI-MS:m/z 485.23(M+H⁺)。

rd5 1- (4-nitrobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 49%, mp:178-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),8.20(d,J＝8.6Hz,2H),7.67(d,J＝8.5Hz,2H),7.51(d,J＝8.6Hz,2H),7.43(t,J＝7.8Hz,2H),7.22–7.09(m,5H),5.72(s,2H).¹³C NMR(100MHz,DMSO)δ158.74(s),157.68(s),156.68(d,J＝4.5Hz),155.08(s),147.38(s),145.22(s),144.60(s),130.58(d,J＝5.9Hz),129.13(s),128.08(s),124.29(s),119.45(d,J＝5.2Hz),97.78(s),49.56(s).ESI-MS:m/z 439.21(M+H⁺)。

rd6 1- (4-aminobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 51%, mp:165-,¹H NMR(400MHz,DMSO)δ8.31(s,1H),8.20(d,J＝8.4Hz,3H),7.69(d,J＝8.3Hz,2H),7.56–7.40(m,5H),7.23–7.09(m,5H),5.73(s,2H).¹³C NMR(100MHz,DMSO)δ158.75,157.70,156.71,156.66,155.09,147.38,145.21,144.62,130.60,130.55,129.18,129.13,128.09,124.29,119.48,119.43,97.80,49.57.ESI-MS:m/z 408.86(M+H⁺)。

rd 7N- (3- ((4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) phenyl) -2-butenamide

White solid, yield 53%,¹H NMR(400MHz,DMSO)δ9.95(s,1H),8.29(s,1H),7.67(d,J＝8.6Hz,2H),7.56(d,J＝8.8Hz,1H),7.43(t,J＝7.9Hz,3H),7.29(t,J＝7.9Hz,1H),7.20-7.11(m,5H),7.05(d,J＝7.7Hz,1H),6.77(dq,J＝13.9,6.8Hz,1H),6.08(d,J＝15.3Hz,1H),5.52(s,2H),1.84(d,J＝6.6Hz,3H).¹³C NMR(100MHz,DMSO)δ163.93(s),158.69(s),157.60(s),156.76(s),156.49(s),154.84(s),144.08(s),140.42(s),140.01(s),138.20(s),132.91(s),130.61,130.55,129.43(s),128.31(s),126.39(s),124.27(s),122.99(s),119.46(s),118.90(s),118.66,97.72(s),50.26(s),17.99(s).ESI-MS:m/z 477.09(M+H⁺)。

rd 81- (4-bromobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 48%, mp:207-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.66(d,J＝8.6Hz,2H),7.53(d,J＝8.3Hz,2H),7.43(t,J＝7.9Hz,2H),7.25(d,J＝8.3Hz,2H),7.19(d,J＝7.4Hz,1H),7.17–7.10(m,4H),5.54(s,2H).¹³C NMR(100MHz,DMSO)δ158.71(s),157.64(s),156.65(d,J＝20.3Hz),154.88(s),144.26(s),137.06(s),131.98(s),130.70–130.20(m),128.21(s),124.28(s),121.25(s),119.45(d,J＝2.4Hz),97.77(s),49.60(s).ESI-MS:m/z472.01(M+H⁺)。

rd9 1- (4- (trifluoromethyl) benzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 62%, mp:182 and 185 deg.C,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.69(dd,J＝15.7,8.1Hz,4H),7.52–7.38(m,4H),7.23–7.08(m,5H),5.67(s,2H).¹³C NMR(100MHz,DMSO)δ158.74(s),157.68(s),156.68(d,J＝12.0Hz),155.03(s),144.44(s),142.35(s),130.58(d,J＝6.1Hz),128.77(s),128.33(d,J＝34.9Hz),126.02(d,J＝3.9Hz),124.29(s),123.29(s),119.46(d,J＝3.5Hz),97.78(s),49.73(s).ESI-MS:m/z 462.31(M+H⁺)。

rd10 1- (3-bromobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 69%, mp:164-,¹H NMR(400MHz,DMSO)δ8.30(s,1H),7.66(d,J＝8.4Hz,2H),7.54–7.47(m,2H),7.43(t,J＝7.1Hz,2H),7.29(dd,J＝17.8,7.2Hz,2H),7.22–7.08(m,5H),5.57(s,2H).¹³C NMR(100MHz,DMSO)δ158.72(s),157.67(s),156.66(d,J＝13.4Hz),154.91(s),144.35(s),140.35(s),131.33(s),130.89(d,J＝16.5Hz),130.57(d,J＝6.8Hz),128.16(s),127.17(s),124.28(s),122.17(s),119.46(d,J＝3.1Hz),97.77(s),49.52(s).ESI-MS:m/z 472.17(M+H⁺)。

rd11 1- (3-chlorobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 66%, mp:162-,¹H NMR(400MHz,DMSO)δ8.30(s,1H),7.66(d,J＝7.2Hz,2H),7.41(dd,J＝22.1,15.1Hz,5H),7.26–7.01(m,6H),5.57(s,2H).¹³C NMR(100MHz,DMSO)δ158.73(s),157.67(s),156.66(d,J＝13.5Hz),154.92(s),144.35(s),140.10(s),133.59(s),131.03(s),130.57(d,J＝6.0Hz),128.57–127.77(m),126.77(s),124.28(s),119.46(d,J＝3.1Hz),97.78(s),49.58(s).ESI-MS:m/z 428.19(M+H⁺)。

rd12 1-benzyl-3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 58%, mp:158-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.66(d,J＝8.5Hz,2H),7.43(t,J＝7.9Hz,2H),7.36–7.25(m,5H),7.19(t,J＝6.0Hz,1H),7.13(t,J＝9.0Hz,4H),5.56(s,2H).¹³C NMR(100MHz,DMSO)δ158.70(s),157.60(s),156.77(s),156.49(s),154.85(s),144.06(s),137.67(s),130.56(d,J＝7.8Hz),129.04(s),128.30(s),128.07(d,J＝5.7Hz),124.27(s),119.40(s),97.75(s),50.26(s).ESI-MS:m/z 394.17(M+H⁺)。

rd13 1- (4-methylbenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 52%, mp:176-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.65(d,J＝8.5Hz,2H),7.43(t,J＝7.9Hz,2H),7.20(d,J＝7.7Hz,3H),7.17–7.10(m,6H),5.50(s,2H),2.25(s,3H).¹³C NMR(100MHz,DMSO)δ158.67(s),157.58(s),156.78(s),156.44(s),154.76(s),143.95(s),137.25(s),134.67(s),130.55(d,J＝8.9Hz),129.56(s),128.24(d,J＝19.0Hz),124.26(s),119.45(s),97.74(s),50.06(s),21.14(s).ESI-MS:m/z 408.18(M+H⁺)。

rd 14- ((4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) benzonitrile 4

White solid, yield 56%, mp:178-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.81(d,J＝8.1Hz,2H),7.67(d,J＝8.5Hz,2H),7.44(t,J＝6.7Hz,4H),7.16(dt,J＝18.2,8.3Hz,5H),5.67(s,2H).¹³C NMR(100MHz,DMSO)δ158.75,157.69,156.72,156.65,155.06,144.54,143.27,133.09,130.63,130.57,128.84,128.12,124.31,119.49,119.45,119.16,110.87,97.78,49.79.ESI-MS:m/z 419.24(M+H⁺)。

rd15 1- (4-Fluorobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 48%, mp:182-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.66(d,J＝8.5Hz,2H),7.44(t,J＝7.9Hz,2H),7.36(dd,J＝8.3,5.7Hz,2H),7.16(dt,J＝18.2,8.3Hz,7H),5.55(s,2H).¹³C NMR(100MHz,DMSO)δ163.25(s),160.83(s),158.70(s),157.62(s),156.89–156.81(m),156.63(d,J＝23.4Hz),154.78(s),144.16(s),133.87(d,J＝3.0Hz),130.56(d,J＝8.0Hz),130.27(d,J＝8.3Hz),128.24(s),124.27(s),119.46(s),115.99(s),115.86(d,J＝21.5Hz),97.76(s),49.51(s).ESI-MS:m/z 412.29(M+H⁺)。

rd16 1- (4-chlorobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 70%, mp:200-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.66(d,J＝8.6Hz,2H),7.47–7.38(m,4H),7.31(d,J＝8.4Hz,2H),7.16(dt,J＝17.6,8.1Hz,5H),5.56(s,2H).¹³C NMR(100MHz,DMSO)δ158.70(s),157.64(s),156.74(s),156.54(s),154.86(s),144.25(s),136.65(s),132.72(s),130.57(d,J＝7.6Hz),130.01(s),129.05(s),128.21(s),124.28(s),119.45(d,J＝2.2Hz),97.76(s),49.53(s).ESI-MS:m/z 428.23(M+H⁺)。

rd17 1- (2-Fluorobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 69%, mp:144-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.65(d,J＝8.6Hz,2H),7.43(t,J＝7.9Hz,2H),7.36(dd,J＝14.0,6.9Hz,1H),7.22(dd,J＝16.5,8.1Hz,3H),7.14(dd,J＝15.9,7.1Hz,5H),5.61(s,2H).¹³C NMR(100MHz,DMSO)δ161.55(s),159.11(s),158.70(s),157.63(s),156.76(s),156.51(s),154.98(s),144.29(s),130.57(d,J＝5.8Hz),128.22(s),125.09(d,J＝3.5Hz),124.34(d,J＝15.9Hz),119.45(d,J＝2.9Hz),116.01(s),115.80(s),97.72(s),43.91(s).ESI-MS:m/z 412.21(M+H⁺)。

rd18 1- (2-chlorobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 55%, mp:148-,¹H NMR(400MHz,DMSO)δ8.29(s,1H),7.67(d,J＝8.5Hz,2H),7.49(d,J＝7.8Hz,1H),7.42(t,J＝7.9Hz,2H),7.29(dt,J＝22.3,7.4Hz,2H),7.21–7.09(m,5H),6.99(d,J＝7.5Hz,1H),5.65(s,2H).¹³C NMR(100MHz,DMSO)δ158.74(s),157.66(s),156.75(s),156.55(s),155.22(s),144.47(s),134.89(s),132.38(s),130.56(d,J＝3.5Hz),129.85(d,J＝4.2Hz),128.20(s),127.93(s),124.25(s),119.44(s),97.75(s),55.38(s),47.77(s).ESI-MS:m/z 428.21(M+H⁺)。

rd19 1- (2-bromobenzyl) -3- (4-phenoxyphenyl) -4-amino-1H-pyrazolo [3,4-d ] pyrimidine

White solid, yield 64%, mp:154-,¹H NMR(400MHz,DMSO)δ8.28(s,1H),7.67(d,J＝8.3Hz,3H),7.43(t,J＝7.7Hz,2H),7.27(dt,J＝25.6,7.2Hz,2H),7.21–7.10(m,5H),6.91(d,J＝7.3Hz,1H),5.62(s,2H).¹³C NMR(100MHz,DMSO)δ158.75(s),157.66(s),156.66(d,J＝19.0Hz),155.25(s),144.51(s),136.49(s),133.14(s),130.58(d,J＝4.0Hz),130.11(s),129.69(s),128.50(s),128.19(s),124.27(s),122.45(s),119.46(d,J＝2.3Hz),97.75(s),50.19(s).ESI-MS:m/z 472.10(M+H⁺)。

15) preparation of target compound Rg

Intermediate 17(0.30g, 1mmol) was dissolved in 3ml DMF and intermediate 10a-10z (1.2mmol), K, was added₂CO₃(0.21g and 1.5mmol), stirring at room temperature for 10h, after the reaction is finished, pouring the reaction liquid into 30ml ice water, extracting with ethyl acetate (30ml multiplied by 3), combining organic phases, washing with saturated saline water, drying, filtering, evaporating the solvent under reduced pressure, and carrying out silica gel column chromatography to obtain the target product Rg, wherein an elution system is dichloromethane/methanol (200: 1-60: 1).

Rg 1:2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-fluorophenyl) acetamide

White solid, yield 61%, mp:216 and 220 deg.C,¹H NMR(400MHz,DMSO)δ10.51(s,1H),8.26(s,1H),7.68(d,J＝8.4Hz,2H),7.61(dd,J＝8.6,5.0Hz,2H),7.44(t,J＝7.8Hz,2H),7.21–7.12(m,7H),5.22(s,2H).¹³C NMR(100MHz,DMSO)δ170.81(s),158.66(s),157.65(s),157.41(s),157.36(s),156.72(s),156.37(s),155.64(s),144.12(s),135.50(s),135.47(s),130.62(s),130.47(s),128.25(s),124.30(s),121.46(s),121.38(s),119.52(s),119.42(s),116.02(s),115.80(s),97.73(s),50.05(s).ESI-MS:m/z 455.21(M+H⁺)。

rg 2:2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-fluorophenyl) acetamide

White solid, yield 58%, mp:242-244 deg.C,¹H NMR(400MHz,DMSO)δ10.49(s,1H),8.26(s,1H),7.68(d,J＝8.4Hz,2H),7.60(dd,J＝8.4,5.0Hz,2H),7.44(t,J＝7.7Hz,2H),7.22–7.11(m,7H),5.22(s,2H).¹³C NMR(100MHz,DMSO)δ170.82(s),159.80(s),158.67(s),157.65(s),157.42(s),156.72(s),156.38(s),155.63(s),144.12(s),135.49(s),135.46(s),130.62(s),130.47(s),128.24(s),124.31(s),121.46(s),121.38(s),119.52(s),119.42(s),116.04(s),115.81(s),97.73(s),50.05(s).ESI-MS:m/z 455.19(M+H⁺)。

rg 3:2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-fluorobenzyl) acetamide

White solid, yield 47%, mp:229-,¹H NMR(400MHz,DMSO)δ8.75(t,J＝5.6Hz,1H),8.26(s,1H),7.67(d,J＝8.4Hz,2H),7.44(t,J＝7.8Hz,2H),7.37(q,J＝7.4Hz,1H),7.22–7.11(m,7H),7.07(t,J＝8.5Hz,1H),5.07(s,2H),4.34(d,J＝5.8Hz,2H).¹³C NMR(100MHz,DMSO)δ167.17(s),163.95(s),161.53(s),158.65(s),157.62(s),156.74(s),156.30(s),155.50(s),144.12(s),142.63(s),142.56(s),130.71(s),130.62(s),130.47(s),128.30(s),124.30(s),123.59(s),123.57(s),119.51(s),119.42(s),114.35(s),114.13(s),97.81(s),49.70(s),42.15(s).ESI-MS:m/z 469.25(M+H⁺)。

rg 4:2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-fluorophenethyl) acetamide

White solid, yield 60%, mp:186-,¹H NMR(400MHz,DMSO)δ8.38-8.33(m,1H),8.25(s,1H),7.66(d,J＝8.4Hz,2H),7.44(t,J＝7.7Hz,2H),7.30–7.03(m,11H),4.96(s,2H),3.22-3.27(m,2H),2.72(t,J＝6.3Hz,2H).¹³C NMR(100MHz,DMSO)δ166.78(s),158.62(s),157.61(s),157.26(s),156.74(s),156.30(s),155.49(s),144.01(s),135.93(s),130.90(s),130.99(s),130.91(s),130.62(s),130.47(s),128.32(s),124.30(s),119.50(s),119.41(s),115.53(s),115.32(s),97.72(s),49.04(s),40.87(s),34.48(s).ESI-MS:m/z 483.22(M+H⁺)。

rg 5:2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chlorophenyl) acetamide

White solid, yield 63%, mp:234-,¹H NMR(400MHz,DMSO)δ10.58(s,1H),8.26(s,1H),7.65(dd,J＝27.0,5.5Hz,4H),7.41(d,J＝18.7Hz,4H),7.28–7.05(m,5H),5.23(s,2H).¹³C NMR(100MHz,DMSO)δ165.80(s),158.67(s),157.66(s),156.72(s),156.39(s),155.64(s),144.15(s),138.03(s),130.62(s),130.47(s),129.26(s),128.23(s),127.64(s),124.31(s),121.21(s),119.62(s),119.53(s),119.42(s),97.72(s),50.12(s).ESI-MS:m/z 471.13(M+H⁺)。

rg 6:2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chloro-4-fluorophenyl) acetamide

White solid, yield 49%, mp:226-,¹H NMR(400MHz,DMSO)δ10.68(s,1H),8.26(s,1H),7.89(d,J＝4.7Hz,1H),7.68(d,J＝8.4Hz,2H),7.48–7.36(m,4H),7.22–7.11(m,5H),5.23(s,2H).ESI-MS:m/z 489.12(M+H⁺)。

rg 7:2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-cyanophenyl) acetamide

White solid, yield 55%, mp:228-,¹H NMR(400MHz,DMSO)δ10.92(s,1H),8.26(s,1H),7.78(dd,J＝17.7,8.6Hz,5H),7.68(d,J＝8.4Hz,2H),7.44(t,J＝7.8Hz,2H),7.23–7.10(m,5H),5.29(s,2H).¹³C NMR(100MHz,DMSO)δ170.84(s),158.67(s),157.67(s),156.71(s),156.57(d,J＝26.8Hz),155.64(s),144.24(s),143.23(s),133.90(s),130.55(d,J＝15.6Hz),128.16(s),124.32(s),119.56(t,J＝15.2Hz),105.87(s),97.71(s),50.23(s).ESI-MS:m/z 462.19(M+H⁺)。

rg 82- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chloro-4-cyanophenyl) acetamide

White solid, yield 61%, mp:219-,¹H NMR(400MHz,DMSO)δ11.10(s,1H),8.26(s,1H),8.02(d,J＝1.8Hz,1H),7.94(d,J＝8.6Hz,1H),7.68(d,J＝8.6Hz,2H),7.61(dd,J＝8.7,1.8Hz,1H),7.44(t,J＝7.9Hz,2H),7.22–7.11(m,5H),5.30(s,2H).¹³C NMR(100MHz,DMSO)δ166.96(s),158.68(s),157.72(s),156.94–156.78(m),156.59(d,J＝24.9Hz),155.71(s),144.31(d,J＝6.6Hz),136.58(s),135.83(s),130.55(d,J＝15.1Hz),128.15(s),124.33(s),119.55(t,J＝13.6Hz),118.37(s),116.62(s),106.37(s),97.75(s),50.28(s).ESI-MS:m/z 496.24(M+H⁺)。

rg 92- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3, 4-dicyanophenyl) acetamide

White solid, yield 46%, mp:228-,¹H NMR(400MHz,DMSO)δ11.24(s,1H),8.29–8.24(m,2H),8.09(d,J＝8.7Hz,1H),8.01–7.93(m,1H),7.68(d,J＝8.5Hz,2H),7.44(t,J＝7.8Hz,2H),7.23–7.09(m,5H),5.32(s,2H).¹³C NMR(100MHz,DMSO)δ167.13(s),158.67(s),157.71(s),157.04–156.72(m),156.57(d,J＝22.3Hz),155.70(s),144.38(s),143.45(s),135.66(s),130.53(d,J＝15.1Hz),128.11(s),124.32(s),123.60(d,J＝9.7Hz),119.47(d,J＝12.7Hz),116.53(s),116.16(d,J＝14.4Hz),108.77(s),97.74(s),50.27(s).ESI-MS:m/z 487.22(M+H⁺)。

rg10 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-cyano-3- (trifluoromethyl) phenyl) acetamide

White solid, yield 48%, mp:226-,¹H NMR(400MHz,DMSO)δ11.27(s,1H),8.27(s,2H),8.13(d,J＝8.5Hz,1H),7.95(d,J＝8.4Hz,1H),7.68(d,J＝8.6Hz,2H),7.44(t,J＝7.9Hz,2H),7.23–7.11(m,5H),5.33(s,2H).¹³C NMR(100MHz,DMSO)δ167.17(s),158.58(s),157.72(s),156.69(s),156.34(s),155.66(s),144.42(s),143.55(s),137.16(s),132.15(s),130.63(s),130.47(s),128.09(s),124.33(s),122.64(s),121.49(s),119.55(s),119.42(s),116.98(s),116.16(s),102.55(s),97.72(s),50.29(s).ESI-MS:m/z 530.21(M+H⁺)。

rg11 4- (2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) acetamido) -2-fluoro-N-methylbenzamide

White solid, yield 66%, mp:241-,¹H NMR(400MHz,DMSO)δ10.94(s,1H),8.26(s,1H),8.09(d,J＝2.9Hz,1H),7.72–7.60(m,4H),7.44(t,J＝7.8Hz,2H),7.36(d,J＝8.3Hz,1H),7.23–7.10(m,5H),5.28(s,2H),2.76(d,J＝4.3Hz,3H).¹³C NMR(100MHz,DMSO)δ166.40(s),163.92(s),161.10(s),158.68(s),158.64(s),157.67(s),156.71(s),156.42(s),155.67(s),144.23(s),142.51(s),142.39(s),131.43(s),131.39(s),130.62(s),130.47(s),128.19(s),124.32(s),119.53(s),119.42(s),118.74(s),118.60(s),115.10(s),115.08(s),106.66(s),106.37(s),97.72(s),50.17(s),26.76(s).ESI-MS:m/z 512.23(M+H⁺)。

rg12 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3- (trifluoromethyl) phenyl) acetamide

White solid, yield 69%, mp:208-,¹H NMR(400MHz,DMSO)δ10.73(s,1H),8.18(d,J＝13.8Hz,1H),8.02(s,1H),7.66(dd,J＝29.1,7.3Hz,3H),7.55–7.47(m,1H),7.37(s,3H),7.19–7.01(m,6H),5.21(s,2H).¹³C NMR(100MHz,DMSO)δ166.33(s),158.69(s),157.69(s),157.18–156.88(m),156.57(d,J＝30.5Hz),155.71(s),144.25(s),139.80(s),130.55(d,J＝14.1Hz),130.47–130.45(m),130.03(d,J＝31.9Hz),128.22(s),125.86(s),124.31(s),123.24(s),120.45(s),119.48(d,J＝10.5Hz),115.74(d,J＝3.8Hz),97.76(s),50.15(s).ESI-MS:m/z 505.15(M+H⁺)。

rg13 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (2-bromophenyl) acetamide

White solid, yield 58%, mp:184-,¹H NMR(400MHz,DMSO)δ9.83(s,1H),8.27(s,1H),7.68(t,J＝7.2Hz,4H),7.47–7.41(m,2H),7.37(t,J＝7.5Hz,2H),7.21–7.12(m,5H),5.29(s,2H).¹³C NMR(100MHz,DMSO)δ166.15(s),158.68(s),157.87(s),157.67(s),156.72(s),156.46(s),155.66(s),144.36(s),136.11(s),133.24(s),130.63(s),130.50(s),128.59(s),128.20(s),127.64(s),126.88(s),124.31(s),119.52(s),119.44(s),97.74(s),49.83(s).ESI-MS:m/z 515.13(M+H⁺)。

rg14 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-bromo-2-methylphenyl) acetamide

White solid, yield 60%，mp:264-268℃，¹H NMR(400MHz,DMSO)δ9.99(s,1H),8.29(s,1H),7.68(d,J＝8.5Hz,2H),7.45(dd,J＝14.3,7.7Hz,3H),7.37(t,J＝8.8Hz,1H),7.23–7.08(m,7H),5.28(s,2H),2.30(s,3H).¹³C NMR(100MHz,DMSO)δ166.09(s),158.63(s),157.66(s),156.70(s),156.34(s),155.59(s),144.21(s),137.55(s),132.68(s),130.55(d,J＝15.4Hz),130.12(s),128.23(s),127.86(s),125.41(d,J＝31.5Hz),124.32(s),119.48(d,J＝10.9Hz),97.78(s),49.83(s),18.68(s).ESI-MS:m/z 529.06(M+H⁺)。

Rg15 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-chloro-2-fluorophenyl) acetamide

White solid, yield 80%, mp:200 and 204 ℃,¹H NMR(400MHz,DMSO)δ10.34(s,1H),8.20(s,1H),7.77(dd,J＝16.4,9.0Hz,1H),7.62(d,J＝8.2Hz,2H),7.37(t,J＝7.6Hz,2H),7.27(t,J＝7.2Hz,1H),7.09(td,J＝12.5,8.2Hz,7H),5.27(s,2H).¹³C NMR(100MHz,DMSO)δ166.43(s),158.68(s),157.69(s),156.87–156.74(m),156.57(d,J＝30.5Hz),155.67(s),144.25(s),130.55(d,J＝13.9Hz),128.22(s),127.71(d,J＝11.6Hz),126.25(s),125.54(d,J＝4.5Hz),124.31(s),123.04(s),120.38(d,J＝15.9Hz),119.48(d,J＝10.5Hz),97.75(s),49.89(s).ESI-MS:m/z489.14(M+H⁺)。

rg16 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-bromo-3- (trifluoromethyl) phenyl) acetamide

White solid, yield 77%, mp:184-,¹H NMR(400MHz,DMSO)δ10.83(s,1H),8.14(dd,J＝31.7,13.4Hz,3H),7.77(d,J＝8.6Hz,1H),7.64(dd,J＝15.5,8.5Hz,4H),7.37(t,J＝7.4Hz,2H),7.17–7.02(m,6H),5.21(s,2H).¹³C NMR(100MHz,DMSO)δ166.46(s),166.06(s),158.68(s),157.70(s),156.71(s),156.44(s),155.70(s),144.29(s),138.94(s),136.14(s),130.54(d,J＝14.1Hz),129.26(s),128.18(s),124.55(s),124.31(s),119.48(d,J＝11.3Hz),118.60(s),112.41(s),97.75(s),50.17(s).ESI-MS:m/z 583.07(M+H⁺)。

rg17 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-nitrophenyl) acetamide

White solid, yield 71%, mp:236-,¹H NMR(400MHz,DMSO)δ10.96(s,1H),8.61(s,1H),8.26(s,1H),7.97–7.89(m,2H),7.72–7.61(m,3H),7.44(t,J＝7.9Hz,2H),7.22–7.11(m,5H),5.29(s,2H).¹³C NMR(100MHz,DMSO)δ166.51(s),158.67(s),157.68(s),156.70(s),156.44(s),155.69(s),148.44(s),144.28(s),140.11(s),130.86(s),130.55(d,J＝15.0Hz),128.17(s),125.65(s),124.32(s),119.48(d,J＝10.5Hz),118.67(s),113.79(s),97.74(s),50.16(s).ESI-MS:m/z 482.22(M+H⁺)。

rg18 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (p-tolyl) acetamide

White solid, yield 69%, mp:222-,¹H NMR(400MHz,DMSO)δ10.27(d,J＝7.4Hz,1H),8.19(s,1H),7.61(d,J＝8.5Hz,2H),7.38(dd,J＝16.4,8.0Hz,5H),7.15–7.09(m,2H),7.08–7.03(m,5H),5.14(s,2H),2.18(s,3H).¹³C NMR(100MHz,DMSO)δ165.34(s),158.66(s),157.65(s),156.73(s),156.36(s),155.64(s),144.08(s),136.59(s),133.00(s),130.55(d,J＝14.8Hz),129.69(s),128.28(s),124.31(s),119.54(t,J＝11.2Hz),97.74(s),50.09(s),20.91(s).ESI-MS:m/z 451.20(M+H⁺)。

rg19 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-cyanophenyl) acetamide

White solid, yield 59%, mp:236-,¹H NMR(400MHz,DMSO)δ10.77(s,1H),8.20(s,1H),7.99(s,1H),7.79–7.71(m,1H),7.62(d,J＝8.6Hz,2H),7.50–7.45(m,2H),7.37(t,J＝7.9Hz,2H),7.15–7.03(m,5H),5.21(s,2H).¹³C NMR(100MHz,DMSO)δ166.37(s),158.68(s),157.69(s),156.72(s),156.43(s),155.70(s),144.26(s),139.83(s),130.87(s),130.55(d,J＝14.3Hz),128.20(s),127.68(s),124.29(d,J＝4.2Hz),122.37(s),119.48(d,J＝11.0Hz),119.06(s),112.16(s),97.75(s),50.13(s).ESI-MS:m/z 462.18(M+H⁺)。

rg20 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (2,3, 4-trifluorophenyl) acetamide

White solid, yield 71%, mp:224-,¹H NMR(400MHz,DMSO)δ10.46(s,1H),8.26(s,1H),7.71–7.58(m,3H),7.44(t,J＝7.9Hz,2H),7.34-7.27(m,1H),7.22–7.11(m,4H),5.31(s,2H).¹³C NMR(100MHz,DMSO)δ166.46(s),158.67(s),157.69(s),156.72(s),156.42(s),155.67(s),144.26(s),138.43(d,J＝14.1Hz),130.55(d,J＝14.0Hz),128.20(s),124.31(s),123.97(d,J＝3.5Hz),119.56(s),119.55–118.71(m),112.37(dd,J＝17.7,3.6Hz),97.75(s),49.75(s).ESI-MS:m/z 491.17(M+H⁺)。

rg21 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4- (trifluoromethyl) phenyl) acetamide

White solid, yield 70%, mp:228-,¹H NMR(400MHz,DMSO)δ10.83(s,1H),8.26(s,1H),7.79(d,J＝8.6Hz,2H),7.75–7.66(m,4H),7.44(t,J＝7.9Hz,2H),7.23–7.11(m,4H),5.28(s,2H).¹³C NMR(100MHz,DMSO)δ166.34(s),158.69(s),157.70(s),156.72(s),156.42(s),155.68(s),144.23(s),142.61(s),130.54(d,J＝13.7Hz),128.22(s),126.67(d,J＝3.6Hz),126.13(s),124.31(s),123.96(s),123.44(s),119.53(t,J＝10.5Hz),97.76(s),50.21(s).ESI-MS:m/z 505.16(M+H⁺)。

rg22 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (2-fluoro-5- (trifluoromethyl) phenyl) acetamide

White solid, yield 59%, mp:158-,¹H NMR(400MHz,DMSO)δ10.93(s,1H),8.26(s,1H),8.19(d,J＝2.1Hz,1H),7.80(dd,J＝8.7,2.1Hz,1H),7.69(t,J＝8.5Hz,3H),7.44(t,J＝7.9Hz,2H),7.22–7.12(m,4H),5.27(s,2H).¹³C NMR(100MHz,DMSO)δ166.46,158.68,157.70,156.72,156.71,156.44,155.70,144.29,138.50,132.76,130.61,130.47,128.18,127.14,124.79,124.79,124.49,124.31,119.53,119.42,118.33,97.75,50.14.ESI-MS:m/z 523.11(M+H⁺)。

rg23 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-nitrophenyl) acetamide

White solid, yield 60%, mp:198-,¹H NMR(400MHz,DMSO)δ11.10(s,1H),8.35–8.21(m,4H),7.83(d,J＝9.0Hz,2H),7.68(d,J＝8.5Hz,2H),7.44(t,J＝7.6Hz,2H),7.22–7.11(m,4H),5.33(s,2H).¹³C NMR(100MHz,DMSO)δ170.83(s),158.67(s),157.69(s),156.70(s),156.45(s),155.67(s),145.13(s),144.28(s),142.97(s),130.55(d,J＝15.2Hz),128.16(s),125.57(s),124.32(s),119.68–119.23(m),97.73(s),50.28(s).ESI-MS:m/z 482.19(M+H⁺)。

rg24 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3, 5-dimethylphenyl) acetamide

White solid, yield 64%, mp:250-,¹H NMR(400MHz,DMSO)δ10.30(s,1H),8.27(s,1H),7.68(d,J＝8.5Hz,2H),7.44(t,J＝7.9Hz,2H),7.23–7.11(m,7H),6.72(s,1H),5.20(s,2H),2.22(s,6H).¹³C NMR(100MHz,DMSO)δ165.47(s),158.65(s),157.62(s),156.73(s),156.35(s),155.67(s),144.07(s),138.95(s),138.27(s),130.55(d,J＝16.4Hz),128.27(s),125.55(s),124.30(s),119.47(d,J＝8.6Hz),117.39(s),97.70(s),50.11(s),21.55(s).ESI-MS:m/z 465.07(M+H⁺)。

rg25 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (4-bromophenyl) acetamide

White solid, yield 62%, mp:238-,¹H NMR(400MHz,DMSO)δ10.71(s,1H),8.25(s,1H),7.68(d,J＝8.6Hz,2H),7.62–7.48(m,5H),7.44(t,J＝7.9Hz,2H),7.24–7.09(m,5H),5.24(s,2H).¹³C NMR(100MHz,DMSO)δ165.86(s),158.65(s),157.64(s),156.71(s),156.38(s),155.63(s),144.12(s),138.58(s),132.09(s),130.63(s),130.47(s),128.22(s),124.31(s),121.55(s),119.52(s),119.42(s),115.58(s),97.69(s),50.11(s).ESI-MS:m/z 515.12(M+H⁺)。

rg26 2- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -N- (3-bromophenyl) acetamide

White solid, yield 74%, mp:188-,¹H NMR(400MHz,DMSO)δ10.64(s,1H),8.26(s,1H),7.93(s,1H),7.68(d,J＝8.6Hz,2H),7.52–7.41(m,3H),7.32-7.26(m,2H),7.22–7.11(m,5H),5.24(s,2H).¹³C NMR(100MHz,DMSO)δ166.06(s),158.67(s),157.66(s),156.72(s),156.41(s),155.66(s),144.19(s),140.60(s),131.39(s),130.63(s),130.47(s),128.21(s),126.72(s),124.32(s),122.09(s),122.00(s),119.53(s),119.43(s),118.42(s),97.71(s),50.12(s).ESI-MS:m/z 515.01(M+H⁺)。

Claims

1,3, 4-trisubstituted pyrazolopyrimidines or pharmaceutically acceptable salts thereof, characterized by having a structure as shown in general formula I:

wherein X is substituted benzyl or substituted acetamido as shown below:

R₂is hydrogen, halogen, C1-6 straight chain or branched chain alkyl, cyano, trifluoromethyl or nitro; r₃Is hydrogen, halogen, C1-6 straight chain or branched chain alkyl, cyano, trifluoromethyl or nitro; r₄Is hydrogen, halogen, C1-6 straight chain or branched chain alkyl; r₆Is hydrogen, C1-6 straight chain or branched chain alkyl, C1-6 straight chain or branched chain substituted acylamino; when X is substituted acetamido, Y is carbon; when X is substituted benzyl, Y is carbon and nitrogen; n is 0,1, 2.

2. The 1,3, 4-trisubstituted pyrazolopyrimidines of claim 1, wherein R is₂Is hydrogen, fluorine, chlorine, bromine, cyano, trifluoromethyl, nitro, methyl; r₃Is hydrogen, fluorine, bromine, cyano, acetyl, methyl, trifluoromethyl, nitro, methylamido; r₄Is hydrogen, fluorine, bromine, methyl; r₆Is hydrogen, fluorine, chlorine, methyl, chloroacetamide, acrylamide, 2-butenamide; when X is substituted acetamido, Y is carbon; when X is substituted benzyl, Y is carbon and nitrogen; n is 0,1, 2.

1,3, 4-trisubstituted pyrazolopyrimidines, characterized in that they are selected from one of the following compounds:

4. use of the 1,3, 4-trisubstituted pyrazolopyrimidines according to any one of claims 1 to 3 for the preparation of an antitumor medicament.

5. The use according to claim 4, for the preparation of a medicament against B-cell malignancies.

6. A pharmaceutical composition against B-cell malignancies comprising a1, 3, 4-trisubstituted pyrazolopyrimidine compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.