CN109608444B

CN109608444B - Isoindolinone-containing ERK inhibitor and preparation method and application thereof

Info

Publication number: CN109608444B
Application number: CN201811424530.3A
Authority: CN
Inventors: 徐云根; 纪德重; 张灵芝; 朱启华; 柏英; 吴尧尧
Original assignee: China Pharmaceutical University
Current assignee: China Pharmaceutical University
Priority date: 2018-11-27
Filing date: 2018-11-27
Publication date: 2022-02-11
Anticipated expiration: 2038-11-27
Also published as: CN109608444A; WO2020107987A1

Abstract

The invention discloses isoindolinone derivatives, in particular compounds containing a 6- (2-aminopyrimidine-4-yl) isoindoline-1-one structure. The invention also discloses a preparation method of the isoindolinone derivative. The invention also discloses the isoindolinone derivatives, stereoisomers, hydrates, solvates or crystals thereof, application of a composition containing the isoindolinone derivatives or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers in preparation of ERK kinase inhibitors, and application of a pharmaceutical composition containing the compounds in preparation of antitumor drugs.

Description

Isoindolinone-containing ERK inhibitor and preparation method and application thereof

Technical Field

The invention belongs to the pharmaceutical chemistry technology, and particularly relates to ERK kinase inhibitors containing 6- (2-aminopyrimidine-4-yl) isoindoline-1-ketone structures, preparation methods thereof, pharmaceutical compositions containing the compounds and application thereof in preparing antitumor drugs.

Technical Field

The Mitogen-activated protein kinases (MAPKs) signal transduction pathway is an important signal pathway for transducing cell surface signals to cell nucleus, and the pathway influences the transcription and regulation of genes in animal cells, thereby causing biological reactions such as cell proliferation, differentiation, transformation, apoptosis and the like. Currently, 5 MAPKs signaling pathways are commonly found in mammalian cells, including Extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinases (JNK/SAPK, c-Jun N-terminal kinase/stress-activated protein kinase), p38 kinase isozymes (p38A, p38B, p38C, and p38D), ERK3/ERK4, and ERK 5.

The RAS-RAF-MEK-ERK signal transduction pathway (ERK pathway) is an evolutionarily conserved signal cascade pathway, which can transfer to cell surface receptor signals, thereby promoting cell proliferation and survival. The ERK signaling pathway plays an important role in maintaining cell stability and regulating cell growth under normal physiological conditions, and is tightly controlled by a multi-stage feedback regulatory pathway.

To date, BRAF inhibitors and MEK inhibitors have met with great success in the anti-tumor field, but with the development of clinical applications, it has been found that most patients will again worsen within one year (acquired resistance) whether BRAF inhibitors are used alone or in combination with MEK inhibitors. In addition, about 10% to 15% of the carrier carries B-Raf^V600EMutant tumor patients are not sensitive to BRAF inhibitors as well as MEK inhibitors (intrinsic resistance). Therefore, the problem of resistance to BRAF and MEK inhibitors is a key scientific problem that needs to be addressed at present.

Compared with the BRAF and MEK inhibitors, the ERK inhibitor firstly blocks the reactivation of the MAPKs expressed by the tumor cells resistant to the BRAF and MEK inhibitors again, and overcomes the existing drug resistance problem. Secondly, tumors inevitably develop drug-resistant mutations, such as the numerous BRAF and MEK mutations mentioned above, over time through drug treatment with targeted kinases. However, to date, little mutation in ERK1/2 was observed in tumor cells. Meanwhile, preclinical experimental results of ERK inhibitors show better effects compared to BRAF and MEK inhibitors. Therefore, compared with inhibiting the upstream kinase of ERK, the ERK inhibitor can block the signal transduction of the ERK pathway more effectively, and overcome the drug resistance of tumor cells to BRAF inhibitors and MEK inhibitors.

Disclosure of Invention

The purpose of the invention is as follows: the invention provides an isoindolinone derivative containing a 6- (2-aminopyrimidine-4-yl) isoindoline-1-one structure, a specific preparation method of the derivative and pharmaceutical application of the derivative in preparing an ERK kinase inhibitor.

The technical scheme is as follows: the invention discloses an isoindolinone derivative shown as a general formula I or pharmaceutically acceptable salt thereof:

wherein:

R¹is selected from

Wherein X is selected from CH₂O, NH or CH-OH; r⁴Selected from H or C1-C6 alkyl, R⁵Selected from H, CH₃、-CH₂OH、-CH₂CH₂OH、-CH₂CH₂CH₂OH or-CH₂CH₂CH₂CH₂OH; y is selected from CH₂O, NH or S; r⁶Selected from F, Cl, Br, CH₃、NH₂Or NHCOCH₃；

R²Is H or-CH₂OH；

R³Is selected from

Wherein R is⁷Is H, F, Cl, Br or OCH₃，R⁷Is mono-, di-or tri-substituted; r⁸Is H or CH₃。

Preferably, R¹Is selected from

Preferably, R³Is selected from

When R is in the specification²is-CH₂OH, the compound I or a pharmaceutically acceptable salt thereof includes chiral isomers:

preferably, the isoindolinone derivatives described herein are selected from the group consisting of I-1 to I-28:

the pharmaceutically acceptable salt is an acid addition salt of the compound shown in the general formula I, wherein the acid for forming the salt is: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.

The compounds of the general formula (I) according to the invention can be prepared by the following process:

the preparation method of the compound of the general formula (I) comprises the following steps:

wherein R is¹Represents:

wherein X represents CH₂、O、CH-OH；R⁴Represents an alkyl group of H, C1 to C6, R⁵Representative H, CH₃、-CH₂OH、-CH₂CH₂OH、-CH₂CH₂CH₂OH or-CH₂CH₂CH₂CH₂OH；R²Represents: h or-CH₂OH；R³Represents:

when R is²＝CH₂OH, compound I configuration includes: s configuration, R configuration and a mixed racemate of the two configurations.

Further, the compound IX is 1-methyl-1H-pyrazol-4-amine, 1-methyl-1H-pyrazol-3-amine, 4-aminotetrahydropyran, 4-aminocyclohexanol, 4-aminopiperidine, 3-aminopyrrolidine, ethanolamine, cyclohexylamine, 3-amino-1-propanol, isopropylamine, 4-aminopyridine; the solvent is selected from sec-butyl alcohol, tert-butyl alcohol, ethanol, tetrahydrofuran, ethyl acetate, methanol or a mixed solvent of any two or three of the solvents, preferably sec-butyl alcohol; the reaction temperature is selected from 90-150 ℃, preferably 110-130 ℃; the reaction time is 4-48 h, and the reaction time is preferably 8-24 h. The compound VIII: the molar ratio of the compound IX is 1: 1-1: 20, preferably 1: 1.5-1: 3.

A second process for the preparation of a compound of formula (I) comprising the steps of:

reacting a compound VIII with tert-butyldimethylsilyl chloride TBSCl to prepare a compound X; then preparing a compound XI from the compound X and IX through substitution reaction; finally, removing a hydroxyl protecting group from a compound XI and then preparing a salt to prepare a compound I.A.

Wherein R is¹Represents:

R³represents:

the configuration of the compound I.A comprises: s configuration, R configuration and a mixed racemate of the two configurations.

It is specifically mentioned that the second preparation process described above is also suitable for R²＝H。

Process for the preparation of compound X by reaction of compound VIII with tert-butyldimethylsilyl chloride (TBSCl): the base is selected from triethylamine, 4-dimethylaminopyridine, imidazole, sodium carbonate or potassium carbonate, preferably imidazole; the reaction solvent is preferably ethyl acetate, tetrahydrofuran, chloroform, acetonitrile, dichloromethane, toluene or a mixed solvent of any two or three of the solvents, preferably dichloromethane; the temperature is selected from 0 ℃ to 50 ℃, preferably 30 ℃ to 40 ℃; the reaction time is selected from 1h to 12h, preferably 7h to 9 h; compound VIII: the molar ratio of tert-butyldimethylsilyl chloride (TBSCl) is 1: 1-1: 10, preferably 1: 1.5-1: 2.

Preparing a compound XI from a compound X and an IX through a substitution reaction, wherein the compound IX is 2-aminothiazole, 2-aminooxazole, 1-methyl-5-aminotetrazole, 1-methyl-1H-3-aminopyrazole or 1-methyl-1H-5-aminopyrazole; the base used is selected from lithium bis (trimethylsilyl) amide, sodium bis (trimethylsilyl) amide or sodium hydride, preferably lithium bis (trimethylsilyl) amide; the solvent is tetrahydrofuran, N-dimethylformamide, dioxane, N-methylpyrrolidone or a mixed solvent of any two or three of the solvents, preferably tetrahydrofuran and N, N-dimethylformamide; the reaction temperature is selected from-80 ℃ to 0 ℃, preferably from-80 ℃ to-25 ℃; the reaction time is selected from 0.1h to 10h, preferably 1h to 3 h; compound X: compound IX: the ratio of the alkali (molar ratio) is 1:1: 1-1: 6:6, preferably 1:1.5: 1.5-1: 2: 2.

The process for preparing the compound I.A by removing a hydroxyl protecting group from a compound XI and then forming a salt, wherein the used deprotection reagent (A) is selected from hydrogen chloride, hydrogen bromide, sulfuric acid or trifluoroacetic acid; the solvent is methanol, ethanol, dichloromethane, acetone, ethyl acetate, tetrahydrofuran or a mixed solvent of any two of the above solvents; the reaction temperature is selected from 0 ℃ to 50 ℃, preferably 0 ℃ to 25 ℃; the reaction time is selected from 0.1h to 6h, preferably 1h to 3 h.

The first preparation method of the key intermediate VIII in the preparation method of the general formula (I) comprises the following steps:

wherein R is²Represents: h or-CH₂OH；R³Represents:

wherein R is⁷Represents H, F, Cl, Br or OCH₃，R⁷May be mono-, di-or tri-substituted; r⁸Represents H or CH₃. When R is²＝CH₂OH, compound VIII configuration includes: s configuration, R configuration and a mixed racemate of the two configurations.

Procedure for preparation of compound IV from compound II and compound III by Suzuki coupling reaction: the catalyst used is selected from palladium tetratriphenylphosphine (Pd (PPh)₃)₄) [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (Pd (dppf) Cl₂) Bis (triphenylphosphine) palladium dichloride (Pd (PPh3)₂Cl₂) Palladium acetate (Pd (OAc)₂) Or (1,1' -bis (diphenylphosphino) ferrocene) nickel dichloride (NiCl)₂(dppf)), preferably palladium tetratriphenylphosphine (Pd (PPh)₃)₄). The base is selected from sodium ethoxide, sodium acetate, potassium phosphate, sodium carbonate or potassium carbonate, preferably sodium carbonate; the reaction solvent is selected from N, N-dimethylformamide, N-dimethylacetamide, ethylene glycol dimethyl ether, 1, 4-dioxane, tetrahydrofuran, toluene, ethanol, water or a mixed solution of any two or three of the solventsAn agent, preferably a toluene/methanol/water mixed solvent; the reaction temperature is selected from 60 ℃ to 120 ℃, and preferably 70 ℃ to 80 ℃; the reaction time is selected from 4 to 24 hours, preferably 7 to 10 hours; compound II: compound III: catalyst: the ratio of the alkali (molar ratio) is 1:1:0.02: 1-6: 1:0.2:6, preferably 1:1:0.03: 1-1: 1.05:0.1: 2.

Process for preparing compound V by oxidation of compound IV: the oxidant is selected from m-chloroperoxybenzoic acid (m-CPBA), hydrogen peroxide, sodium hypochlorite, periodic acid, sodium periodate, potassium permanganate or tert-butyl peroxy alcohol, preferably m-chloroperoxybenzoic acid; the reaction solvent is selected from tetrahydrofuran, acetone, dichloromethane, 1, 4-dioxane or acetonitrile, preferably dichloromethane; the reaction temperature is selected from 10 ℃ to 80 ℃, preferably 20 ℃ to 30 ℃; the reaction time is selected from 0.5 h-5 h, preferably 1 h-2 h; compound IV: the molar ratio of the oxidant is 1: 1-1: 10, preferably 1: 1.5-1: 3.

Process for preparing compound VI from compound V by bromination: the brominating agent used is selected from N-bromosuccinimide (NBS) or liquid bromine, preferably N-bromosuccinimide; the solvent is selected from benzene, carbon tetrachloride, chloroform, acetonitrile, dichloromethane, methanol or toluene, preferably carbon tetrachloride; the reaction temperature is selected from 50 ℃ to 100 ℃, and preferably 70 ℃ to 90 ℃; the reaction time is selected from 1h to 18h, preferably 4h to 7 h; compound V: the ratio of the brominating agent (molar ratio) is 1: 1-1: 20, preferably 1: 1-1: 2.

Process for preparing compound VIII by cyclization of compound VI with compound VII: the base used is selected from triethylamine, benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBop), N-Diisopropylethylamine (DIEA), sodium carbonate or potassium carbonate, preferably triethylamine; the reaction solvent is selected from tetrahydrofuran, acetone, acetonitrile, methanol, dichloromethane or a mixed solvent of any two, preferably acetonitrile; the reaction temperature is selected from 50 ℃ to 100 ℃, preferably 60 ℃ to 90 ℃; the reaction time is selected from 1h to 24h, preferably 5h to 9 h; the molar ratio of the compound VI to the compound VII to the base is 1:1: 1-1: 4:8, preferably 1:1: 1-1: 1.5: 1.5.

The second preparation method of the key intermediate VIII in the preparation method of the general formula (I) comprises the following steps:

wherein R is²Represents: h; r³Represents:

wherein R is⁷Represents H, F, Cl, Br or OCH₃，R⁷May be mono-, di-or tri-substituted; r⁸Represents H or CH₃。

Procedure for preparation of compound XIII from compound II and compound XII by Suzuki coupling reaction: the reaction is carried out under the condition of adding a catalyst, a base and a reaction solvent, wherein the catalyst is selected from bis (triphenylphosphine) palladium dichloride, tetrakis (triphenylphosphine) palladium or [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride, preferably tetrakis (triphenylphosphine) palladium; the base is selected from sodium ethoxide, sodium acetate, potassium phosphate, sodium carbonate or potassium carbonate, preferably sodium carbonate; the reaction solvent is selected from N, N-dimethylformamide, N-dimethylacetamide, ethylene glycol dimethyl ether, 1, 4-dioxane, tetrahydrofuran, toluene, methanol, ethanol, water or a mixed solvent of any two or three of the solvents, preferably a mixed solvent of toluene/methanol/water; the reaction temperature is selected from 50 ℃ to 120 ℃, preferably 60 ℃ to 80 ℃; the reaction time is selected from 3 to 30 hours, preferably 7 to 12 hours; compound II: compound XII: catalyst: the ratio of the alkali (molar ratio) is 1:1:0.02: 1-6: 1:0.3:10, preferably 1:1:0.05: 1-1: 1.05:0.1: 2.

Process for the preparation of compound XIV from compound XIII by deprotection with a deprotecting agent selected from hydrogen chloride, trifluoroacetic acid or boron trifluoride, preferably trifluoroacetic acid; the reaction solvent is selected from tetrahydrofuran, acetone, dichloromethane, 1, 4-dioxane or acetonitrile, preferably dichloromethane; the reaction temperature is selected from 0 ℃ to 50 ℃, preferably 20 ℃ to 30 ℃; the reaction time is selected from 0.5h to 6h, preferably 1h to 2 h.

Process for the preparation of Compound XVI by substitution reaction of Compound XIV and Compound XV: the base is selected from potassium carbonate, sodium carbonate, potassium phosphate, potassium acetate, potassium hydroxide, sodium hydride or sodium acetate, preferably sodium hydride; the reaction solvent is selected from toluene, methanol, water, 1, 4-dioxane, glycol dimethyl ether or a mixed solvent of any three of the above, preferably 1, 4-dioxane; the reaction temperature is selected from 0 ℃ to 120 ℃, preferably 50 ℃ to 80 ℃; the reaction time is selected from 3 to 24 hours, preferably 9 to 12 hours; the molar ratio of the compound XIV to the compound XV to the base is selected from 1:1:1 to 1:6:6, preferably 1:1:1 to 1:1.2: 1.5.

Procedure for preparation of compound VIII by oxidation of compound XVI: the oxidant is selected from m-chloroperoxybenzoic acid (m-CPBA), hydrogen peroxide, sodium hypochlorite, periodic acid, sodium periodate, potassium permanganate or tert-butyl peroxy alcohol, preferably m-chloroperoxybenzoic acid; the reaction solvent is selected from tetrahydrofuran, acetone, dichloromethane, 1, 4-dioxane or acetonitrile, preferably dichloromethane; the reaction temperature is selected from 10 ℃ to 80 ℃, preferably 20 ℃ to 30 ℃; the reaction time is selected from 0.1h to 10h, preferably 1h to 2 h; compound IV: the proportion of the oxidant (molar ratio) is 1: 1-1: 6, preferably 1: 1.5-1: 3.

The invention also discloses a pharmaceutical composition which contains the compound of the general formula I or pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The compound can be added with pharmaceutically acceptable carriers to prepare common pharmaceutical preparations, such as tablets, capsules, syrup and suspending agents, and common pharmaceutical excipients such as spices, sweeteners, liquid or solid fillers or diluents and the like can be added.

The application of the isoindolinone derivative shown in the general formula I and the stereoisomer, hydrate, solvate or crystal thereof in preparing ERK kinase inhibitor drugs is also within the protection scope of the invention.

Furthermore, the ERK kinase inhibitor is used for preparing the medicine for treating malignant tumors.

Has the advantages that: the invention discloses a novel isoindolinone derivative shown in a general formula (I), and pharmacological experiments show that the compound I can generate good inhibition effect on ERK kinase and can be used for preparing a medicament for treating malignant tumors with over-activated ERK pathways; the invention also discloses a preparation method of the isoindolinone derivative.

Drawings

FIG. 1 shows the results of HCT116 nude mouse transplantable tumor experiments with Compound I-8.

Detailed Description

The present application will be described in detail with reference to specific examples.

Example 1

Synthesis of 2- (3-chlorobenzyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-1)

Synthesis of tert-butyl 6- (2- (methylthio) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylate (XIII)

4-chloro-2- (methylthio) pyrimidine (XII) (6.14g,38.24mmol), tert-butyl 6- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) isoindoline-2-carboxylate (II) (13.74g,38.24mmol), sodium carbonate (8.11g,76.48mmol) and tetratriphenylphosphine palladium (2.21g,1.91mmol) were dissolved in a mixed solvent composed of toluene (150mL), ethanol (50mL) and water (50mL), reacted at 85 ℃ for about 12h under nitrogen protection, after TLC (dichloromethane: methanol ═ 35:1) detection of reaction completion, cooled to room temperature, evaporated to dryness under reduced pressure, the residue was slurried with ethyl acetate (300mL), filtered, the filter cake was washed with ethyl acetate and separated by column chromatography (eluent: dichloromethane: methanol: 100: 1), 12.3g of solid yellow soil is obtained, the yield is 90.0%, and m.p.197.7-199.2 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.60(d,J＝5.2Hz,1H,ArH),8.53(s,1H,ArH),8.47(dd,J＝8.0,1.6Hz,1H,ArH),7.61(d,J＝8.0Hz,1H,ArH),7.44(d,J＝5.3Hz,1H,ArH),4.84(s,2H,CH₂),2.65(s,3H,SCH₃),1.61(s,9H,3×CH₃).

Synthesis of 6- (2- (methylthio) pyrimidin-4-yl) isoindolin-1-one (XIV)

Dissolving compound XIII (8.0g,22.4mmol) in dichloromethane (80mL), adding trifluoroacetic acid (20mL), stirring at room temperature for about 1h, stopping the reaction until the starting material reaction is complete as detected by TLC (dichloromethane: methanol ═ 25:1), evaporating the solvent under reduced pressure, adding water (100mL) to the residue, adjusting pH to 6-7 with saturated sodium hydroxide solution, precipitating a white solid, filtering, and drying to obtain 5.5g of a white solid, yield: 95.5%, m.p. >250.0 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.69(s,2H,ArH),8.44(s,1H,ArH),7.88(s,1H,ArH),7.74(d,J＝7.8Hz,1H,ArH),4.47(s,2H,CH₂),2.61(s,3H,CH₃).

Synthesis of 2- (3-chlorobenzyl) -6- (2- (methylthio) pyrimidin-4-yl) isoindolin-1-one (XVI-1)

Dissolving compound XIV (2g,7.8mmol) in 1, 4-dioxane (50mL), adding 60% sodium hydride (376mg,9.4mmol), stirring at room temperature for 1h, adding 3-chlorobenzyl bromide (2.4g,11.7mmol), stirring at 70 ℃ for reaction for about 12h, detecting by TLC (petroleum ether: ethyl acetate ═ 2:1) that the reaction of the raw material XIV is complete, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: petroleum ether: ethyl acetate ═ 6: 1) to obtain 1.93g of yellow solid, yield: 64.8 percent and m.p.152.2-153.4 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.61(d,J＝5.3Hz,1H,ArH),8.55(d,J＝1.0Hz,1H,ArH),8.40(dd,J＝8.0,1.6Hz,1H,ArH),7.55(d,J＝8.0Hz,1H,ArH),7.47(d,J＝5.3Hz,1H,ArH),7.34-7.27(m,3H,ArH),7.25-7.17(m,1H,ArH),4.83(s,2H,CH₂),4.37(s,2H,CH₂),2.66(s,3H,SCH₃).

Synthesis of 2- (3-chlorobenzyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-1)

Dissolving compound XVI-1(1.0g,2.6mmol) in dichloromethane (20mL), adding m-chloroperoxybenzoic acid (1.31g,6.5mmol) under ice bath, stirring at room temperature for about 2h, detecting complete reaction of the raw material XVI-1 by TLC (dichloromethane: methanol ═ 35:1), adding saturated sodium thiosulfate solution (30mL) to the reaction solution, stirring vigorously for 10min, standing for layering, extracting the aqueous layer with dichloromethane (30mLx3), combining the organic layers, washing with saturated sodium carbonate solution (30mLx3), washing with saturated sodium chloride solution (30mL), drying with anhydrous sodium sulfate, filtering, and evaporating the solvent under reduced pressure to obtain white solid 890mg, yield: 82.7 percent and m.p.145.3-146.1 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.99(d,J＝5.3Hz,1H,ArH),8.59(d,J＝0.6Hz,1H,ArH),8.53(dd,J＝8.0,1.5Hz,1H,ArH),8.03(d,J＝5.3Hz,1H,ArH),7.62(d,J＝8.0Hz,1H,ArH),7.36-7.28(m,3H,ArH),7.26-7.19(m,1H,ArH),4.83(s,2H,CH₂),4.41(s,2H,CH₂),3.46(s,3H,SO₂CH₃).

Dissolving compound VIII-1(200mg,0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg,0.73mmol) in sec-butanol (3mL), heating to 125 ℃ with a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the raw materials are completely reacted, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1) to obtain 116mg of yellow solid, yield: 55.6 percent and m.p.181.9-183.0 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.49(s,1H,ArH),8.37(d,J＝4.0Hz,1H,ArH),8.27(d,J＝7.6Hz,1H,ArH),7.51(d,J＝7.9Hz,1H,ArH),7.35-7.25(m,3H,ArH),7.21(d,J＝3.4Hz,1H,ArH),7.06(d,J＝4.7Hz,1H,ArH),5.37-5.22(m,1H,NH),4.81(s,2H,CH₂),4.35(s,2H,CH₂),4.26-4.10(m,1H,NHCH),4.04-4.00(m,2H,OC ₂H),3.63-3.56(m,2H,OC ₂H),2.12-2.08(m,2H,NHCHC ₂H),1.66-1.56(m,2H,NHCHC ₂H).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.10,163.96,161.79,158.75,143.23,138.92,133.06,130.89,130.32,130.18,128.18,128.03,126.27,123.25,122.58,114.84,106.71,66.80,49.51,47.22,46.02,33.29.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₄ClN₄O₂:435.1582；Found:435.1590.

IR(cm^-1):3434.79,3253.87,1703.98,1600.62,1572.80,1531.13,1415.70,1261.52,1197.50,1137.32,803.41,765.66,709.03,611.12.

Example 2

Synthesis of 2- (3-fluoro-4-chlorobenzyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-2) Synthesis of 2- (4-chloro-3-fluorobenzyl) -6- (2- (methylthio) pyrimidin-4-yl) isoindolin-1-one (XVI-2)

Dissolving compound XIV (1g,3.9mmol) in 1, 4-dioxane (50mL), adding 60% sodium hydride (188mg,4.7mmol), stirring at room temperature for 1h, adding 4-chloro-3-fluorobenzyl bromide (1.3g,5.85mmol), stirring at 70 ℃ for about 12h, detecting the completion of the reaction of the raw material XIV by TLC (petroleum ether: ethyl acetate ═ 2:1), evaporating the solvent under reduced pressure, and separating the residue by column chromatography (eluent: petroleum ether: ethyl acetate ═ 6: 1) to obtain 855mg of white solid, yield: 54.8 percent and m.p.143.2-144.5 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.63(d,J＝5.2Hz,1H,ArH),8.57(s,1H,ArH),8.43(dd,J＝8.0,1.6Hz,1H,ArH),7.58(d,J＝8.0Hz,1H,ArH),7.49(d,J＝5.3Hz,1H,ArH),7.41(t,J＝7.8Hz,1H,ArH),7.12(ddd,J＝9.5,8.9,1.6Hz,2H,ArH),4.83(s,2H,CH₂),4.40(s,2H,CH₂),2.70(s,3H,SCH₃).

Synthesis of 2- (4-chloro-3-fluorobenzyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-2)

Dissolving compound XVI-2(500mg,1.3mmol) in dichloromethane (20mL), adding m-chloroperoxybenzoic acid (657mg,3.25mmol) under ice bath, stirring at room temperature for about 2h, after TLC (dichloromethane: methanol ═ 35:1) detects that the raw material-2 is completely reacted, adding saturated sodium thiosulfate solution (20mL) into the reaction solution, stirring vigorously for 10min, standing for layering, extracting the aqueous layer with dichloromethane (20mLx3), combining the organic layers, washing with saturated sodium carbonate (20mLx3), washing with saturated sodium chloride solution (20mL), drying with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure to dryness to obtain white solid 447mg, yield: 79.6 percent and m.p.153.8-155.2 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.99(d,J＝5.3Hz,1H,ArH),8.58(s,1H,ArH),8.53(d,J＝7.9Hz,1H,ArH),8.03(d,J＝5.3Hz,1H,ArH),7.62(d,J＝8.0Hz,1H,ArH),7.38(t,J＝7.9Hz,1H,ArH),7.13(d,J＝9.1Hz,1H,ArH),7.07(d,J＝8.2Hz,1H,ArH),4.81(s,2H,CH₂),4.41(s,2H,CH₂),3.45(s,3H,SO₂CH₃).

Synthesis of 2- (3-fluoro-4-chlorobenzyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-2)

Dissolving compound VIII-2(207mg,0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg,0.73mmol) in sec-butanol (3mL), heating to 125 ℃ with a sealed tube to react for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the raw materials are completely reacted, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1) to obtain 115mg of white solid, yield: 52.9 percent and m.p.180.4-181.9 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.49(s,1H,ArH),8.37(d,J＝3.8Hz,1H,ArH),8.28(dd,J＝8.0,1.5Hz,1H,ArH),7.52(d,J＝7.9Hz,1H,ArH),7.37(t,J＝7.8Hz,1H,ArH),7.19-6.99(m,3H,ArH),5.30(brs,1H,NH),4.80(s,2H,CH₂),4.36(s,2H,C ₂H),4.26-4.11(m,1H,NHCH),4.04-4.00(m,2H,OC ₂H),3.63-3.56(m,2H,OC ₂H),2.12-2.08(m,2H,NHCHC ₂H),1.68-1.55(m,2H,NHCHC ₂H).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.11,164.03,160.77(d,J＝124.1Hz),158.52,156.63,143.19,137.86(d,J＝10.9Hz),133.16,132.92,131.04,130.45,124.39(d,J＝3.7Hz),123.29,122.64,116.26(d,J＝21.3Hz),115.00,106.65,66.78,49.50,47.25,45.66,33.26.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₃ClFN₄O₂:453.1488；Found:453.1494.

IR(cm^-1):3427.73,3248.25,2922.60,2843.90,1701.76,1602.04,1570.09,1528.32,1414.52,1365.96,1162.42,1196.23,1055.28,867.41,802.31,768.02,612.67,535.91.

Example 3

Synthesis of 2- (3-chloro-4-fluorobenzyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-3)

Synthesis of 2- (3-chloro-4-fluorobenzyl) -6- (2- (methylthio) pyrimidin-4-yl) isoindolin-1-one (XVI-3)

Dissolving compound XIV (1g,3.9mmol) in 1, 4-dioxane (50mL), adding 60% sodium hydride (188mg,4.7mmol), stirring at room temperature for 1h, adding 4-fluoro-3-chlorobenzyl bromide (1.3g,5.85mmol), stirring at 70 ℃ for about 12h, detecting the completion of the reaction of the raw material XIV by TLC (petroleum ether: ethyl acetate ═ 2:1), evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: petroleum ether: ethyl acetate ═ 6: 1) to obtain 933mg of orange solid, yield: 59.8 percent and m.p.174.9-176.3 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.60(d,J＝5.0Hz,1H,ArH),8.54(s,1H,ArH),8.40(d,J＝8.0Hz,1H,ArH),7.55(d,J＝7.9Hz,1H,ArH),7.46(d,J＝5.2Hz,1H,ArH),7.38(d,J＝6.8Hz,1H,ArH),7.21(brs,1H,ArH),7.12(t,J＝8.5Hz,1H,ArH),4.78(s,2H,CH₂),4.37(s,2H,CH₂),2.67(s,3H,SCH₃).

Synthesis of 2- (3-chloro-4-fluorobenzyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-3)

Dissolving compound XVI-3(500mg,1.25mmol) in dichloromethane (20mL), adding m-chloroperoxybenzoic acid (657mg,3.25mmol) under ice bath, stirring at room temperature for about 2h, after TLC (dichloromethane: methanol ═ 35:1) detects that the raw material has reacted completely, adding saturated sodium thiosulfate solution (20mL) to the reaction solution, stirring vigorously for 10min, standing for layering, extracting the aqueous layer with dichloromethane (20mLx3), combining the organic layers, washing with saturated sodium carbonate (20mLx3), washing with saturated sodium chloride solution (20mL), drying with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure to dryness to obtain white solid 446mg, yield: 82.6 percent and m.p.173.2-174.4 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.99(d,J＝5.3Hz,1H,ArH),8.57(s,1H,ArH),8.53(dd,J＝8.0,1.7Hz,1H,ArH),8.02(d,J＝5.3Hz,1H,ArH),7.62(d,J＝8.0Hz,1H,ArH),7.39(dd,J＝6.8,2.1Hz,1H,ArH),7.26-7.18(m,1H,ArH),7.13(t,J＝8.6Hz,1H,ArH),4.80(s,2H,CH₂),4.40(s,2H,CH₂),3.46(s,3H,SO₂CH₃).

Dissolving compound VIII-3(207mg,0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg,0.73mmol) in sec-butanol (3mL), heating to 125 ℃ with a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-3 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1) to obtain 104mg of white solid, yield: 47.8 percent and m.p.210.5-211.7 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.49(s,1H,ArH),8.37(d,J＝5.2Hz,1H,ArH),8.27(dd,J＝8.0,1.4Hz,1H,ArH),7.52(d,J＝7.8Hz,1H,ArH),7.38(dd,J＝6.8,1.9Hz,1H,ArH),7.23-7.19(m,1H,ArH),7.16-7.01(m,2H,ArH),5.30(s,1H,NH),4.78(s,2H,CH₂),4.35(s,2H,CH₂),4.26-4.09(m,1H,NHCH),4.04-4.00(m,2H,OC ₂H),3.63-3.56(m,2H,OC ₂H),2.12-2.08(m,2H,NHCHC ₂H),1.68-1.55(m,2H,NHCHC ₂H).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.07,164.00,163.26,160.49(d,J＝171.3Hz),158.58,143.19,137.92,134.01(d,J＝3.6Hz),132.99,130.40,130.29,127.95,127.86,123.28,122.62,116.99(d,J＝21.3Hz),106.65,66.78,49.45,47.24,45.49,33.27.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₃ClFN₄O₂:453.1488；Found:453.1492.

IR(cm^-1):3469.03,3249.92,2924.39,2847.62,1701.05,1604.79,1571.85,1531.09,1493.95,1261.83,1238.03,868.32,803.01,761.81,628.16.

Example 4

Synthesis of 2- (3-chlorobenzyl) -6- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-4)

Compound VIII-1(200mg,0.48mmol) and 1-methyl-1H-pyrazol-5-amine (IX-2) (71mg,0.73mmol) were dissolved in tetrahydrofuran (5mL), cooled to-25 ℃, 1N LiHMDS (0.73mL,0.73mmol) was added, reacted at-25 ℃ for about 3H, after TLC (dichloromethane: methanol ═ 25:1) detection that the raw material VIII-1 reacted completely, the solvent was evaporated under reduced pressure, and the residue was separated by column chromatography (dichloromethane: methanol ═ 150: 1) to give 52mg of a white solid, yield: 25.1 percent and m.p.167.9-169.2 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.58-8.46(m,2H,ArH),8.30(d,J＝7.3Hz,1H,ArH),7.61-7.46(m,2H,ArH),7.41-7.16(m,6H,ArH,NH),6.39(s,1H,ArH),4.81(s,2H,CH₂),4.36(s,2H,CH₂),3.84(s,3H,CH₃).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.43,164.92,160.79,159.57,144.19,139.27,138.84,137.77,137.51,135.25,133.62,130.96,130.63-130.53(m),128.66,128.55,126.75,124.01,123.04,109.59,99.89,50.02,46.50,36.07.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₃H₂₀ClN₆O:431.1382；Found:431.1389.

IR(cm^-1):3446.17,3227.14,1680.22,1581.58,1554.06,1446.36,1408.42,1263.38,1200.56,812.52,769.37,709.28,682.87,609.28.

Example 5

Synthesis of 2- (3-chlorobenzyl) -6- (2- ((1-methyl-1H-pyrazol-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-5)

Dissolving compound VIII-1(200mg,0.48mmol) and 1-methyl-1H-pyrazol-4-amine (IX-3) (71mg,0.73mmol) in sec-butanol (3mL), heating to 125 ℃ in a sealed tube to react for about 12H, detecting completion of the reaction of VIII-1 by TLC (dichloromethane: methanol ═ 25:1), cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (dichloromethane: methanol ═ 150: 1) to obtain 107mg of a light yellow solid, yield: 51.7 percent and m.p.160.1-161.3 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.56(s,1H,NH),8.5-8.47(m,2H,ArH),8.37(d,J＝8.1Hz,1H,ArH),7.90(s,1H,ArH),7.72(d,J＝7.6Hz,1H,ArH),7.57(s,1H,ArH),7.45-7.34(m,4H,ArH),7.27-7.25(m,1H,ArH),4.77(s,2H,CH₂),4.48(s,2H,CH₂),3.82(s,3H,CH₃).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):167.58,163.42,160.26,159.87,144.74,140.50,133.78,133.19,131.12,130.50,129.40,128.04,127.88,126.88,124.67,123.68,121.69,120.95,117.71,107.29,50.00,45.44,39.26.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₃H₂₀ClN₆O:431.1382；Found:431.1386.

IR(cm^-1):3446.28,1682.78,1625.78,1573.42,1433.39,1199.84,798.84,769.01,710.30,611.06.

Example 6

Synthesis of 2- (3-chloro-4-fluorobenzyl) -6- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-6)

Dissolving compound VIII-3(207mg,0.48mmol) and 1-methyl-1H-pyrazol-5-amine (IX-2) (71mg,0.73mmol) in tetrahydrofuran (5mL), cooling to-25 ℃, adding 1N LiHMDS (0.73mL,0.73mmol), reacting at-25 ℃ for about 3 hours, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-3 is complete, evaporating the solvent under reduced pressure, and performing column chromatography (dichloromethane: methanol ═ 150: 1) to obtain a light yellow solid 62mg, yield: 28.8 percent and m.p.108.1-109.2 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.55-8.45(m,2H,ArH),8.29(d,J＝8.0Hz,1H,ArH),7.54-7.46(m,2H,ArH),7.38(d,J＝6.2Hz,1H,ArH),7.30(d,J＝4.7Hz,1H,ArH),7.20(brs,1H,NH),7.15-7.09(m,1H,ArH),7.02(s,1H,ArH),6.37(s,1H,ArH),4.78(s,2H,CH₂),4.36(s,2H,CH₂),3.83(s,3H,CH₃).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):167.94,164.44,160.22,159.86(d,J＝92.7Hz),159.03,143.64,138.32,137.27,137.06,133.87(d,J＝3.8Hz),133.05,130.55,130.30,127.96,127.87,123.55,122.60,117.01(d,J＝21.3Hz),109.13,99.50,49.50,45.49,35.56.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₃H₁₉ClFN₆O:449.1287；Found:449.1290.

IR(cm^-1):3431.63,1685.96,1583.06,1554.62,1499.00,1445.94,1405.63,1315.27,1247.74,1200.54,1133.99,816.71,764.94,609.57,541.08.

Example 7

Synthesis of 2- (1- (3, 4-dichlorophenyl) -2-hydroxyethyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-7)

Synthesis of methyl 2-methyl-5- (2- (methylthio) pyrimidin-4-yl) benzoate (IV)

Dissolving 4-chloro-2- (methylthio) pyrimidine (XII) (19.8g,123.4mmol), methyl 2-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzoate (III) (34.1g,123.4mmol), tetratriphenylphosphine palladium (7.1g,6.17mmol) and sodium carbonate (26.2g,246.8mmol) in a mixed solvent composed of toluene (90mL), methanol (30mL) and water (30mL), reacting at 70 ℃ for about 8h under the protection of nitrogen, allowing TLC (petroleum ether: ethyl acetate: 10:1) to detect that the raw materials are completely reacted, cooling to room temperature, adding ethyl acetate (100mL), standing for layering, evaporating the solvent under reduced pressure to obtain a residue, separating by column chromatography (petroleum ether: ethyl acetate: 25:1) to obtain 29.8g of a white solid, yield: 88.0 percent and m.p.66.2-67.3 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.58(d,J＝1.9Hz,1H,ArH),8.54(d,J＝5.3Hz,1H,ArH),8.16(dd,J＝8.0,2.0Hz,1H,ArH),7.40-7.35(m,2H,ArH),3.92(s,3H,OCH₃),2.65(s,3H,SCH₃),2.64(s,3H,CH₃).

Synthesis of methyl 2-methyl-5- (2- (methylsulfonyl) pyrimidin-4-yl) benzoate (V)

Dissolving compound IV (24.2g,88.3mmol) in dichloromethane (150mL), adding m-chloroperoxybenzoic acid (38.1g,220.8mmol) in portions under ice bath, stirring at room temperature for about 2h, detecting that the raw material IV completely reacts by TLC (dichloromethane: methanol ═ 20:1), adding dichloromethane (100mL) for dilution, adding saturated sodium thiosulfate solution (100mL), stirring for 20min, standing for layering, extracting an aqueous layer with dichloromethane (100mL × 3), combining organic layers, washing with saturated sodium carbonate solution (100mL), washing with saturated sodium chloride solution (100mL), drying with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure to obtain 25.7g of light yellow solid, yield: 95.0 percent and m.p.121.8-123.3 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.91(d,J＝5.3Hz,1H,ArH),8.62(d,J＝1.9Hz,1H,ArH),8.23(dd,J＝8.1,2.0Hz,1H,ArH),7.93(d,J＝5.3Hz,1H,ArH),7.41(d,J＝8.1Hz,1H,ArH),3.93(s,3H,OCH₃),3.42(s,3H,SCH₃),2.66(s,3H,CH₃).

Synthesis of methyl 2- (bromomethyl) -5- (2- (methylsulfonyl) pyrimidin-4-yl) benzoate (VI)

The starting material V (23.6g,77.0mmol) and NBS (15.1g,84.7mmol) were dissolved in benzene (150mL), azobisisobutyronitrile (2.5g,15.4mmol) was added and reacted at 80 ℃ for about 3h, TLC (petroleum ether: ethyl acetate 1:1) checked for completion of the starting material reaction, cooled to room temperature, concentrated to a red oil and put into the next step without further treatment.

Synthesis of 2- (1- (3, 4-dichlorophenyl) -2-hydroxyethyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-4)

Compound VI (1g,2.6mmol), 2-amino-2- (3, 4-dichlorophenyl) ethyl-1-ol (VII-1) (533mg,2.6mmol) and triethylamine (540 μ L,3.9mmol) were dissolved in methanol (10mL), refluxed for about 6 hours, TLC (dichloromethane: methanol ═ 20:1) checked for completion of the starting material reaction, cooled to below 10 ℃ in an ice bath, stirred for 30min, solid precipitated, and suction filtered to give 694mg of white solid, yield: 55.8 percent and m.p.203.9-204.9 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.12(d,J＝5.1Hz,1H,ArH),8.59(s,1H,ArH),8.52(d,J＝8.4Hz,1H,ArH),8.39(d,J＝4.8Hz,1H,ArH),7.87(d,J＝8.5Hz,1H,ArH),7.69-7.56(m,2H,ArH),7.39(d,J＝9.6Hz,1H,ArH),5.50-5.37(m,1H,HOCH₂CH),4.82(d,J＝19.0Hz,1H,CONC ₂H),4.59(d,J＝18.4Hz,1H,CONC ₂H),4.24-4.02(m,2H,HOC ₂H),3.55(s,3H,SO₂C ₃H).

Dissolving compound VIII-4(229mg,0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg,0.73mmol) in sec-butanol (3mL), heating to 125 ℃ in a sealed tube to react for about 12h, detecting complete reaction by TLC (dichloromethane: methanol ═ 25:1), cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (dichloromethane: methanol ═ 250: 1-50: 1) to obtain 124mg of white solid, yield: 51.7 percent and m.p.217.9-219.3 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.42(s,1H,ArH),8.39(d,J＝5.2Hz,1H,ArH),8.34(d,J＝8.3Hz,1H,ArH),7.72(d,J＝8.2Hz,1H,ArH),7.64(d,J＝1.4Hz,1H,ArH),7.61(d,J＝8.4Hz,1H,ArH),7.35(dd,J＝8.3,1.4Hz,1H,ArH),7.31(s,1H,NH),7.26(d,J＝5.3Hz,1H,ArH),5.36(t,J＝6.6Hz,1H,HOCH₂CH),4.72(d,J＝17.9Hz,1H,CONC ₂H),4.48(d,J＝18.2Hz,1H,CONC ₂H),4.12-3.96(m,3H,HOC ₂H,NHCH),3.91-3.88(m,2H,O(C ₂ ₂H)),3.59-3.40(m,2H,O(C ₂ ₂H)),1.61-1.57(m,2H,NHC ₂H),1.51-1.51(m,2H,NHC ₂H).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):167.41,163.63,161.62,158.95,144.38,139.78,137.03,132.67,131.16,130.66,130.07,129.86,129.50,128.02,123.92,120.96,105.66,66.09,60.84,55.84,47.28,46.77,32.46.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₅Cl₂N₄O₃:499.1298；Found:499.1295.

IR(cm^-1):3427.73,3298.57,2939.08,2839.35,1567.99,1524.11,1455.27,1411.81,1161.99,1137.06,1084.73,871.24,802.94,772.14,606.59,524.48.

Example 8

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-8)

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-5)

Compound VI (9.63g,25.0mmol), 2-amino-2- (3-chlorophenyl) ethyl-1-ol (VII-2) (4.29g,25.0mmol) and triethylamine (10.4mL,74.9mmol) were dissolved in methanol (100mL), refluxed for about 6 hours, and after completion of the reaction of the starting materials by TLC (dichloromethane: methanol ═ 20:1), cooled in ice bath to 10 ℃ or less, stirred for 30min, solid precipitated, and suction filtered to obtain 7.2g of a white solid, yield: 64.9 percent and m.p.166.2-167.4 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):9.01(d,J＝5.3Hz,1H,ArH),8.49(s,1H,ArH),8.44(dd,J＝8.0,1.5Hz,1H,ArH),8.01(d,J＝5.3Hz,1H,ArH),7.58(d,J＝8.0Hz,1H,ArH),7.50-7.31(m,3H,ArH),7.28-7.26(m,1H,ArH),5.40(dd,J＝7.9,4.4Hz,1H,CHCH₂OH),4.63(d,J＝17.8Hz,1H,C ₂H),4.43-4.32(m,2H,HOC ₂HCH),4.28(dd,J＝11.9,4.4Hz,1H,C ₂H),3.49(s,3H,SO₂C ₃H).

Dissolving compound VIII-5(213mg,0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg,0.73mmol) in sec-butanol (3mL), heating to 125 ℃ with a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol: 25:1) that the reaction of the raw material VIII-5 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and performing column chromatography (dichloromethane: methanol: 250: 1-50: 1) to obtain 117mg of a light yellow solid, yield: 52.4 percent and m.p.141.0-142.3 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.47(s,1H,ArH),8.34-8.26(m,2H,ArH),7.54-7.51(m,1H,ArH),7.34-7.30(m.3H,ArH),7.24-7.21(m,1H,ArH),7.11-7.05(m,1H,ArH),5.30-5.24(m,1H,HOCH₂CH),4.51(d,J＝17.0Hz,1H,CONC ₂H),4.37-4.19(m,4H,CONC ₂H,HOC ₂H,NHCH),4.04-4.01(m,2H,O(C ₂ ₂H)),3.63-3.57(m,2H,O(C ₂ ₂H)),2.11-2.08(m,2H,NHC ₂H),1.68-1.64(m,2H,NHC ₂H).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):169.71,165.54,162.32,159.43,143.86,139.89,138.43,135.35,133.52,130.93,130.75,128.89,128.09,126.24,123.60,122.91,107.12,67.31,63.65,60.08,49.66,47.68,33.76.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₆ClN₄O₃:465.1688；Found:465.1688.

IR(cm^-1):3421.83,3266.10,2946.95,2851.70,1677.23,1571.70,1454.84,1454.84,1203.44,1084.97,802.24,771.67,521.54.

Example 9

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-9)

Synthesis of 2- (2- ((tert-butyldimethylsilyl) oxy) -1- (3-chlorophenyl) ethyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (X-1)

Dissolving compound VIII-5(1.0g,2.26mmol) and imidazole (0.31g,4.52mmol) in dichloromethane (15mL), adding TBSCl (0.68g,4.52mmol), stirring at 35 ℃ for about 8h, detecting the complete reaction of the raw material VIII-5, washing with saturated sodium bicarbonate (6mL), washing with saturated sodium chloride solution (6mL), drying with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure, adding petroleum ether (6mL), pulping, and filtering to obtain a white solid 1.20g, yield 95.1%, m.p.131.7-133.2 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.99(d,J＝5.3Hz,1H,ArH),8.58-8.49(m,2H,ArH),8.02(d,J＝5.4Hz,1H,ArH),7.63(d,J＝7.9Hz,1H,ArH),7.44(s,1H,ArH),7.29(d,J＝6.5Hz,3H,ArH),5.58(t,J＝5.4Hz,1H,CHCH₂OSi),4.70(d,J＝17.9Hz,1H,C ₂H),4.36(d,J＝18.0Hz,1H,C ₂H),4.28-4.21(m,2H,SiOC ₂HCH),3.46(s,3H,SO₂C ₃H),0.84(s,9H,SiC(C ₃ ₃H)),0.07(s,3H,SiC ₃H),0.05(s,3H,SiC ₃H).

Dissolving a compound X-1(200mg,0.36mmol) and 1-methyl-1H-pyrazol-5-amine (IX-2) (52mg,0.54mmol) in anhydrous tetrahydrofuran (5mL), cooling to-25 ℃, adding 1N LiHMDS (0.54mL,0.54mmol), reacting at 25 ℃ for about 3H, detecting that the raw material X-1 is completely reacted by TLC (dichloromethane: methanol ═ 25:1), evaporating the solvent under reduced pressure, adding ethyl acetate (5mL), dissolving, adjusting the pH to 2-3 by using an ethyl acetate solution of HCl, continuously stirring for about 3H, detecting that the intermediate is completely reacted by TLC (dichloromethane: methanol ═ 25:1), evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1), so as to obtain 35mg of a white solid, wherein the yield: 21.1 percent and m.p.118.2-119.4 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.45(d,J＝5.0Hz,1H,ArH),8.32(s,1H,ArH),8.07(d,J＝7.5Hz,1H,ArH),7.84(s,1H,ArH),7.41-7.35(m,3H,ArH),7.28(m,2H,ArH),7.17(d,J＝5.0Hz,1H,ArH),6.15(s,1H,ArH),5.41(s,1H,HOCH₂CH),4.52(d,J＝17.4Hz,1H,CONC ₂H),4.47-4.12(m,3H,CONC ₂H,HOC ₂H),3.81(s,3H,CH₃).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):169.37,165.04,161.12,159.75,144.28,139.92,138.64,137.93,137.60,135.33,133.56,130.91,130.71,128.84,128.18,126.33,123.75,122.87,109.67,100.15,63.15,59.25,49.13,36.09.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₂ClN₆O₂:461.1487；Found:461.1489.

IR(cm^-1):3411.10,3233.04,2938.05,1673.72,1588.57,1554.41,1446.65,1406.90,1267.97,1202.18,815.17,770.16,610.50.

Example 10

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((1-methyl-1H-pyrazol-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-10)

Dissolving compound VIII-5(160mg,0.36mmol) and 1-methyl-1H-pyrazol-4-amine (IX-3) (52mg,0.54mmol) in sec-butyl alcohol (3mL), heating to 125 ℃ in a sealed tube, reacting for about 12H, detecting complete reaction by TLC (dichloromethane: methanol 25:1), cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol 200: 1-50: 1) to obtain an earthy yellow solid 93mg, yield: 56.0 percent and m.p.218.2-219.4 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.59(s,1H,NH),8.82-8.20(m,3H,ArH),7.91(s,1H,ArH),7.75(s,1H,ArH),7.56(s,1H,ArH),7.50-6.91(m,5H,ArH),5.38(s,1H,OH),5.21(s,1H,HOCH₂CH),4.73(d,J＝16.7Hz,1H,CONC ₂H),4.47(d,J＝17.8Hz,1H,CONC ₂H),4.01(s,2H,HOC ₂H),3.82(s,3H,CH₃).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):167.88,163.44,160.25,159.86,144.96,141.58,133.72,133.36,130.96,130.52,130.45,127.96,127.73,126.74,124.61,123.68,121.57,120.93,111.49,107.28,61.41,56.74,47.67,39.27.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₂ClN₆O₂:461.1487；Found:461.1494.

IR(cm^-1):3427.46,3261.74,3062.46,2920.09,1680.07,1626.00,1573.30,1451.93,1433.52,1333.65,1200.93,991.89,802.54,769.16,617.20.

Example 11

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (isopropylamino) pyrimidin-4-yl) isoindolin-1-one (I-11)

Dissolving compound VIII-5(200mg,0.45mmol) and isopropylamine (IX-3) (40mg,0.68mmol) in sec-butyl alcohol (3mL), sealing the tube, heating to 125 ℃ for reaction for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-5 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 96mg of white solid, yield: 50.4 percent and m.p.140.1-142.4 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.42(s,1H,ArH),8.36-8.32(m,1H,ArH),8.24(d,J＝7.6Hz,1H,ArH),7.46-7.44(m,1H,ArH),7.33-7.29(m.3H,ArH),7.26-7.23(m,1H,ArH),6.98(d,J＝5.2Hz,1H,ArH),5.33-5.26(m,2H,HOCH₂CH,NHCH),4.52(d,J＝17.5Hz,1H,CONC ₂H),4.31-4.25(m,4H,CONC ₂H,HOC ₂H,NHCH),1.30(d,J＝6.5Hz,6H,NHCH ₃ ₂(CH)).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):158.27,142.81,138.92,137.47,134.37,132.40,130.01,129.79,127.91,127.10,125.28,122.86,122.56,122.05,121.86,110.05,105.54,62.46,58.94,48.58,42.33,22.40.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₃H₂₄ClN₄O₂:423.1588；Found:423.1588.

IR(cm^-1):3478.65,3245.37,1660.99,1597.27,1573.23,1537.51,1459.55,1408.28,1203.39,1178.35,1177.90,773.23.

Example 12

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (2-hydroxyethylamino) pyrimidin-4-yl) isoindolin-1-one (I-12)

Dissolving compound VIII-5(300mg,0.68mmol) and ethanolamine (IX-5) (124mg,2.03mmol) in sec-butyl alcohol (3mL), heating to 125 ℃ in a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that raw material VIII-5 completely reacts, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain white solid 160mg, yield: 55.7 percent and m.p.157.5-160.1 ℃.

¹H-NMR(300MHz,CD₃OD)δ(ppm):8.51(s,1H,ArH),8.35-8.32(m,2H,ArH),7.66-7.64(m,1H,ArH),7.44-7.34(m.4H,ArH),7.17-7.16(m,1H,ArH),5.53-5.49(m,1H,HOCH₂CH),4.80-4.72(m,1H,CONC ₂H),4.44(d,J＝18.1Hz,1H,CONC ₂H),4.27-4.17(m,2H,HO ₂CHCH),3.77-3.76(m,2H,HO ₂CHCH₂NH),3.67-3.62(m,2H,HOCH₂ ₂CHNH).

¹³C-NMR(75MHz,CD₃OD)δ(ppm):169.02,163.52,161.88,157.75,143.99,139.34,136.96,133.85,131.92,129.97,129.67,127.34,126.86,125.21,122.97,121.03,105.35,60.49,59.98,56.40,42.78,25.31.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₂H₂₂ClN₄O₃:425.1380；Found:425.1376.

IR(cm^-1):3253.11,1655.42,1624.36,1575.99,1545.27,1458.44,1403.24,1360.94,1272.46,1203.64,1201.80,1065.99.

Example 13

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (3-hydroxypropylamino) pyrimidin-4-yl) isoindolin-1-one (I-13)

Dissolving compound VIII-5(300mg,0.68mmol) and propanolamine (IX-6) (152mg,2.03mmol) in sec-butyl alcohol (3mL), sealing the tube, heating to 125 ℃ for reaction for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that raw material VIII-5 completely reacts, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 130mg of white solid, yield: 43.6 percent and m.p.64.2-66.3 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.37(s,1H,ArH),8.28(s,1H,ArH),8.17(d,J＝7.7Hz,1H,ArH),7.46(d,J＝7.7Hz,1H,ArH),7.35-7.26(m,4H,ArH),6.98-6.96(m,1H,ArH),5.99(s,1H,HNCH₂),5.44-5.40(m,1H,HOCH₂ CH),4.59(d,J＝17.5Hz,1H,CONC ₂H),4.38-4.21(m,3H,CONC ₂H,HO ₂CHCH),4.02(brs,2H,HOCH₂CH,HOCH₂CH₂),3.75-3.68(m,4H,HO ₂CHCH₂ ₂CHNH),1.88-1.85(m,2H,HOCH₂ ₂CHCH₂NH).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.75,163.48,162.27,158.06,143.11,138.85,136.96,134.34,132.47,129.88,129.78,127.89,127.11,125.31,122.72,121.96,105.98,62.08,58.26,48.17,37.08,32.37,26.46.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₃H₂₄ClN₄O₃:439.1537；Found:439.1532.

IR(cm^-1):3439.85,1668.07,1572.74,1493.88,1454.98,1407.24,1342.27,1266.70,1201.84,1067.90,806.12,770.92,695.77,613.93.

Example 14

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (cyclohexylamino) pyrimidin-4-yl) isoindolin-1-one (I-14)

Dissolving compound VIII-5(400mg,0.90mmol) and cyclohexylamine (IX-7) (268mg,2.70mmol) in sec-butyl alcohol (3mL), heating to 125 ℃ in a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that raw material VIII-5 completely reacts, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 180mg of white solid, yield: 43.2 percent and m.p.150.4-151.8 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.37(s,1H,ArH),8.25(d,J＝7.8Hz,1H,ArH),8.16(s,1H,ArH),7.56(d,J＝7.9Hz,1H,ArH),7.40(s,1H,ArH),7.32-7.29(m,3H,ArH),7.06(d,J＝5.7Hz,1H,ArH),5.50-5.46(m,1H,HOCH₂ CH),4.73(d,J＝17.7Hz,1H,CONC ₂H),4.45-4.23(m,4H,CONC ₂H,HO ₂CHCH,CH₂ CHNH),4.02(s,1H,HOCH₂CH),2.11-2.04(m,2H,cyclohexyl-H),1.91-1.83(m,2H,cyclohexyl-H),1.68-1.36(m,6H,cyclohexyl-H).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.08,164.51,157.71,143.69,138.87,135.44,134.33,132.83,130.25,129.78,128.51,127.89,127.16,125.29,123.00,122.35,104.63,61.86,57.74,49.82,47.92,32,02,25.02,24.07

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₆H₂₈ClN₄O₂:463.1901；Found:463.1896.

IR(cm^-1):3297.74,2928.85,1673.28,1570.45,1524.32,1455.75,1419.22,1283.98,1205.37,798.16,770.61,709.39,682.87.

Example 15

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((4-hydroxycyclohexyl) amino) pyrimidin-4-yl) isoindolin-1-one (I-15)

Dissolving compound VIII-5(400mg,0.90mmol) and 4-aminocyclohexanol (IX-8) (311mg,2.70mmol) in sec-butanol (3mL), sealing the tube and heating to 125 ℃ for reaction for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-5 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 210mg of white solid, yield: 48.7 percent and m.p.170.3-171.6 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.42(s,1H,ArH),8.32-8.31(m,1H,ArH),8.24(d,J＝8.0Hz,1H,ArH),7.49(d,J＝8.0Hz,1H,ArH),7.35(s,1H,ArH),7.32-7.26(m,3H,ArH),7.01(d,J＝5.4Hz,1H,ArH),5.73(brs,1H,HNCH),5.40-5.36(m,1H,HOCH₂ CH),4.57(d,J＝17.5Hz,1H,CONCH ₂),4.38-4.22(m,3H,CONCH ₂,HO ₂CHCH),3.99-3.96(m,1H,(CH₂)₂ CHOH),3.78-3.68(m,1H,(CH₂)₂ CHNH),2.23-2.05(m,4H,hydroxycyclohexyl-H),1.59-1.33(m,4H,hydroxycyclohexyl-H).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.57,153.71,143.49,138.88,136.74,134.34,132.54,130.03,129.78,128.51,127.89,127.13,127.03,125.29,122.75,122.01,105.55,69.32,62.17,58.21,48.94,48.13,33.39,30.13.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₆H₂₈ClN₄O₃:479.1850；Found:479.1846.

IR(cm^-1):3429.06,2929.86,2848.98,1654.59,1594.29,1575.12,1524.49,1495.10,1456.75,1414.30,1347.78,1203.31,1068.26,808.64,771.78,689.80,612.24.

Example 16

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((tetrahydrofuran-3-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-16)

Dissolving compound VIII-5(400mg,0.90mmol) and 3-aminotetrahydrofuran (IX-8) (48mg,1.35mmol) in sec-butanol (3mL), heating to 125 ℃ with a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-5 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 220mg of white solid, yield: 54.2 percent and m.p.192.3-193.9 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.38-8.34(m,2H,ArH),8.20(d,J＝7.9Hz,1H,ArH),7.43(d,J＝7.7Hz,1H,ArH),7.32-7.28(m,3H,ArH),7.24-7.23(m,1H,ArH),7.03-7.02(m,1H,ArH),5.72(s,1H,NHCH),5.35-5.30(m,1H,HOCH₂CH),4.70-4.68(m,1H,NHCH),4.54(d,J＝17.6Hz,1H,CONCH ₂),4.34-4.20(m,3H,CONCH ₂,HO ₂CH),4.05-4.01(m,2H,O ₂CH),3.91-3.89(m,1H,OCH ₂),3.80-3.78(m,1H,OCH ₂),2.40-2.34(m,1H,CHCH ₂CH₂O),1.98-1.94(m,1H,CHCH ₂CH₂O).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):169.04,163.03,161.58,158.33,142.98,138.89,137.27,134.35,132.41,129.94,129.78,127.89,127.10,125.29,122.59,121.83,106.20,73.27,66.61,62.23,58.43,51.51,48.27,32.81.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₄ClN₄O₃:451.1537；Found:451.1533.

IR(cm^-1):3239.95,1669.85,1566.82,1532.89,1458.47,1426.89,1407.90,1334.69,1200.00,800.56,771.43,685.71,612.24.

Example 17

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (2-aminothiazole) pyrimidin-4-yl) isoindolin-1-one (I-17)

Dissolving 2-aminothiazole (IX-10) (36mg,0.36mmol) in anhydrous tetrahydrofuran (5mL), cooling to-78 ℃, adding 1N LiHMDS (0.36mL,0.36mmol), stirring for 30min, dissolving compound X-1(100mg,0.18mmol) in anhydrous tetrahydrofuran (3mL), slowly dropwise adding the reaction solution at-78 ℃ to react at-78 ℃ for about 1h, detecting the completion of the reaction of raw material X-1 by TLC (dichloromethane: methanol: 20:1), evaporating the solvent under reduced pressure, adding ethyl acetate (5mL) to dissolve, adjusting the pH to 2-3 with an ethyl acetate solution of HCl, continuing stirring for about 3h, detecting the intermediate reaction by TLC (dichloromethane: methanol: 15: 1), evaporating the solvent under reduced pressure, separating by column chromatography (eluent: dichloromethane: methanol: 1-30: 1), yield 45mg of the target compound hydrochloride as a golden yellow solid: 50.0%, m.p. >220.1 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.73-8.72(m,1H,ArH),8.61(s,1H,ArH),8.55-8.52(m,1H,ArH),7.82-7.76(m,2H,ArH),7.55-7.50(m,1H,ArH),7.43-7.36(m,4H,ArH),7.23(s,1H,ArH),5.39-5.32(m,1H,HOCH₂ CH),4.76(d,J＝17.9Hz,1H,CONCH ₂),4.49(d,J＝18.2Hz,1H,CONCH ₂),4.07-4.01(m,2H,HO ₂CHCH).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):158.69,155.65,152.61,148.10,136.88,132.54,127.24,126.17,124.71,124.55,121.96,120.09,118.97,118.74,117.74,115.83,113.39,106.05,103.98,101.06,52.36,47.83,38.78.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₃H₁₉ClN₅O₂S:464.0948；Found:464.0947.

IR(cm^-1):3404.23,1666.47,1549.64,1453.87,1395.88,1203.23,1166.75,1072.97,771.35,691.31,614.43.

Example 18

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((1-methyl-1H-pyrazol-3-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-18)

Dissolving N-methyl-3-aminopyrazole (IX-11) (35mg,0.36mmol) in anhydrous tetrahydrofuran (5mL), cooling to-78 ℃, adding 1N LiHMDS (0.36mL,0.36mmol), stirring for 30min, dissolving compound X-1(100mg,0.18mmol) in anhydrous tetrahydrofuran (3mL), slowly dropwise adding the reaction solution at-78 ℃ to react at-78 ℃ for about 1h, detecting by TLC (dichloromethane: methanol ═ 20:1) that raw material X-1 has completely reacted, evaporating the solvent under reduced pressure, adding ethyl acetate (5mL) to dissolve, adjusting pH to 2-3 with HCl ethyl acetate solution, stirring for about 3h, detecting by TLC (dichloromethane: methanol ═ 15: 1) that the intermediate has completely reacted, evaporating the solvent under reduced pressure, separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-30: 1), yield 50mg of the hydrochloride of the target compound as a white solid: 55.8 percent and m.p.218.2-219.8 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.50-8.49(m,1H,ArH),8.44(s,1H,ArH),8.38-8.36(m,1H,ArH),7.75-7.72(m,1H,ArH),7.58(s,1H,ArH),7.46-7.35(m,5H,ArH),6.64(s,1H,ArH),5.38-5.33(m,1H,HOCH₂ CH),5.24(s,1H,HNC),4.72(d,J＝18.7Hz,1H,CONCH ₂),4.44(d,J＝18.6Hz,1H,CONCH ₂),4.04-3.97(m,2H,HO ₂CHCH),3.74(s,3H, ₃CHN).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):159.21,154.35,151.09,150.89,139.21,135.97,132.21,128.26,124.74,124.10,122.47,122.05,121.59,119.06,118.61,117.53,115.67,112.53,99.15,88.07,83.25,52.13,47.79,38.66

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₂ClN₆O₂:461.1493；Found:461.1483.

IR(cm^-1):3416.33,1660.65,1569.89,1404.85,1204.08,1073.47,795.92,771.43,608.16.

Example 19

Synthesis of 2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (1-methyl-1H-tetrazol-5-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-19)

Dissolving a compound X-1(500mg,0.90mmol) and 1-methyl-5-aminotetrazole (IX-12) (228mg,2.30mmol) in anhydrous N, N-dimethylformamide (4mL), cooling to-70 ℃, adding 1N LiHMDS (1.35mL,1.35mmol), reacting at 70 ℃ for about 2 hours, detecting the reaction by TLC (dichloromethane: methanol ═ 20:1), evaporating the solvent under reduced pressure, adding ethyl acetate (5mL), dissolving, adjusting the pH to 2-3 by using an ethyl acetate solution of HCl, continuously stirring for about 3 hours, detecting the reaction completion of an intermediate by TLC (dichloromethane: methanol ═ 15: 1), evaporating the solvent under reduced pressure, and carrying out column chromatography (eluent: dichloromethane: methanol ═ 200: 1-30: 1) to obtain a light yellow solid, wherein the target product hydrochloride is 180mg, and the yield: 40.1 percent and m.p.86.3-88.0 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.62-8.60(m,1H,ArH),8.43(s,1H,ArH),8.35-8.32(m,1H,ArH),7.75-7.73(m,2H,ArH),7.42-7.32(m,4H,ArH),5.39-5.35(m,1H,HOCH₂ CH),5.19(s,1H,HNCH),4.73(d,J＝18.2Hz,1H,CONCH ₂),4.45(d,J＝18.2Hz,1H,CONCH ₂),4.08-4.02(m,2H,HO ₂CHCH),3.91(s,3H,N ₃CH)

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):159.11,154.89,151.34,150.70,142.71,136.40,132.10,127.27,124.75,124.16,122.04,121.62,119.08,118.58,117.51,115.78,112.65,101.76,52.09,47.78,38.66,25.12

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₂H₂₀ClN₈O₂:463.1398；Found:463.1397.

IR(cm^-1):3417.98,1664.86,1626.84,1594.43,1556.37,1495.65,1439.49,1410.86,1372.67,1227.31,1204.91,1061.42,822.97,770.34,686.55.

Example 20

Synthesis of (R) -2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-20)

Synthesis of (R) -2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-6)

Dissolving compound VI (1.57g,4.08mmol), (R) -2-amino-2- (3-chlorophenyl) ethyl-1-ol (VII-3) (700mg,4.08mmol) and triethylamine (1.70mL,12.24mmol) in acetonitrile (30mL), heating under reflux for about 6h, evaporating the solvent under reduced pressure after the reaction is detected by TLC (dichloromethane: methanol ═ 20:1), adding methanol (10mL), cooling in ice bath to below 10 ℃, stirring for 30min, precipitating a solid, and performing suction filtration to obtain 1.03g of a white solid, yield: 56.9 percent and m.p.78.2-79.8 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.15(d,J＝5.3Hz,1H,ArH),8.60-8.55(m,3H,ArH),7.84(d,J＝8.0,1H,ArH),7.44(s,1H,ArH),7.38-7.32(m,3H,ArH),5.41-5.37(m,1H,CHCH₂OH),5.22-5.19(m,1H,CHCH₂ OH),4.78(d,J＝18.6Hz,1H,CH ₂N),4.51(d,J＝18.6Hz,1H,CH ₂N)4.08 4.01(m,2H,HO ₂CHCH),3.51(s,3H, ₃CH).

Dissolving compound VIII-6(400mg,0.90mmol) and 4-aminotetrahydropyran (IX-1) (182mg,1.80mmol) in sec-butanol (5mL), heating to 125 ℃ with a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-6 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 208mg of white solid, yield: 49.7 percent and m.p.83.8-85.7 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.47-8.37(m,3H,ArH),7.77(d,J＝7.8Hz,1H,ArH),7.48(s,1H,ArH),7.42-7.37(m,3H,ArH),7.30-7.29(m,1H,ArH),5.45-5.41(m,1H,HOCH₂ CH),5.27-5.24(m,1H,HOCH₂CH),4.78(d,J＝18.3Hz,1H,CONC ₂H),4.50(d,J＝18.3Hz,1H,CONC ₂H),4.09-4.07(m,3H,NHCH,HO ₂CH),3.96-3.93(m,2H,O(C ₂ ₂H)),3.56-3.47(m,2H,O(C ₂ ₂H)),1.96-1.92(m,2H,NHCH(C ₂ ₂H)),1.66-1.59(m,2H,NHCH(C ₂ ₂H)).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):167.91,162.73,161.52,159.11,144.21,140.61,136.99,133.28,132.43,130.54,130.03,127.58,127.10,125.98,124.01,120.93,105.74,66.07,60.63,56.34,47.19,46.61,32.29.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₆ClN₄O₃:465.1693；Found:465.1687.

IR(cm^-1):3415.44,2949.76,2359.85,1673.13,1573.18,1524.05,1454.41,1408.20,1201.03,1136.01,1077.26,806.04,767.72,605.57.

Example 21

Synthesis of (R) -2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((1-methyl-1H-pyrazol-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-21)

Dissolving compound VIII-6(400mg,0.90mmol) and 1-methyl-1H-pyrazol-4-amine (IX-3) (175mg,1.80mmol) in sec-butyl alcohol (5mL), heating to 125 ℃ in a sealed tube to react for about 12H, detecting by TLC (dichloromethane: methanol: 20:1) that the reaction of the raw material VIII-6 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol: 200: 1-50: 1) to obtain a golden yellow solid 100mg, wherein the yield is: 24.1 percent and m.p.229.7-231.6 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.61(s,1H,NH),8.58-8.56(m,1H,ArH),8.52(s,1H,ArH),8.45-8.43(m,1H,ArH),7.97(s,1H,ArH),7.83-7.80(m,1H,ArH),7.64(s,1H,ArH),7.49-7.39(m,5H,ArH),5.47-5.42(m,1H,HOCH₂ CH),5.27-5.24(m,1H,HOCH₂CH),4.79(d,J＝18.0Hz,1H,CONCH ₂),4.52(d,J＝18.2Hz,1H,CONCH ₂),4.14-4.07(m,2H,HO ₂CH),3.88(s,3H,N ₃CH).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):167.80,163.00,159.62,159.37,144.39,140.69,136.89,133.27,132.59,130.54,130.12,129.99,127.57,127.12,126.04,124.18,122.93,121.03,120.75,106.93,60.67,56.33,47.20,38.58.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₂ClN₆O₂:461.1493；Found:461.1492.

IR(cm^-1):3421.26,3262.27,1674.63,1626.32,1572.86,1454.67,1433.10,1405.87,1333.36,1299.81,1201.52,1039.41,992.27,803.19,768.62,615.10.

Example 22

Synthesis of (S) -2- (2-hydroxy-1-phenylethyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-22)

Synthesis of (S) -2- (2-hydroxy-1-phenylethyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-7)

Dissolving compound VI (2.81g,7.29mmol), L-phenylglycinol (VII-4) (1g,7.29mmol) and triethylamine (2.03mL,14.58mmol) in acetonitrile (30mL), heating and refluxing for about 6h, detecting by TLC (dichloromethane: methanol 20:1) that the raw materials have reacted, evaporating the solvent under reduced pressure, adding methanol (10mL), cooling in ice bath to below 10 ℃, stirring for 30min, precipitating solids, and performing suction filtration to obtain 1.65g of white solid, wherein the yield is: 55.3 percent and m.p.154.2-155.8 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.13-9.12(m,1H,ArH),8.56-8.53(m,3H,ArH),7.81(d,J＝7.8Hz,1H,ArH),7.34-7.26(m,5H,ArH),5.41-5.37(m,1H,CHCH₂OH),5.16-5.14(m,1H,CHCH₂ OH),4.75(d,J＝18.7Hz,1H,CH ₂N),4.43(d,J＝18.5Hz,1H,CH ₂N),4.06-3.95(m,2H,HO ₂CHCH),3.48(s,3H,SO₂ ₃CH).

Dissolving compound VIII-7(400mg,0.98mmol) and 4-aminotetrahydropyran (IX-1) (198mg,1.96mmol) in sec-butanol (5mL), sealing the tube and heating to 125 ℃ for reaction for about 12h, detecting by TLC (dichloromethane: methanol ═ 20:1) that the reaction of the raw material VIII-7 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 260mg of white solid, yield: 61.6 percent and m.p.143.9-145.6 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.42-8.31(m,3H,ArH),7.71(d,J＝8.0Hz,1H,ArH),7.35-7.24(m,6H,ArH),5.42-5.34(m,1H,HOCH₂ CH),5.17-5.13(m,1H,HOCH₂CH),4.71(d,J＝18.2Hz,1H,CONC ₂H),4.40(d,J＝18.1Hz,1H,CONC ₂H),4.09-3.98(m,3H,HO ₂CHCH,HNCH),3.91-3.88(m,2H,CH₂ CH ₂OCH ₂CH₂),3.44-3.42(m,2H,CH₂ CH ₂OCH ₂CH₂),1.91-1.85(m,2H,CH₂ CH ₂CHCH ₂CH₂),1.57-1.53(m,2H,CH₂ CH ₂CHCH ₂CH₂).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):168.23,162.07,159.67,144.66,138.63,137.45,133.24,130.36,129.12,128.08,127.74,126.10,124.46,121.37,106.18,66.58,61.30,57.14,47.38,47.11,32.84.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₇N₄O₃:431.2083；Found:431.2073.

IR(cm^-1):3376.06,3252.41,2932.72,2845.54,1676.71,1567.26,1526.08,1453.81,1410.60,1364.66,1305.55,1274.66,1201.98,1136.46,1088.16,769.49,703.07.

Example 23

Synthesis of (S) -2- (2-hydroxy-1-phenylethyl) -6- (2- ((1-methyl-1H-pyrazol-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-23)

Dissolving compound VIII-7(400mg,0.98mmol) and 1-methyl-1H-pyrazol-4-amine (IX-3) (190mg,1.96mmol) in sec-butyl alcohol (5mL), heating to 125 ℃ in a sealed tube to react for about 12H, detecting by TLC (dichloromethane: methanol ═ 20:1) that the reaction of the raw material VIII-7 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 206mg of white solid, yield: 49.3 percent and m.p.227.9-229.7 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.58(s,1H,HN),8.52-8.51(m,1H,ArH),8.46(s,1H,ArH),8.38(d,J＝8.4Hz,1H,ArH),7.91(s,1H,ArH),7.76(d,J＝8.1Hz,1H,ArH),7.57(s,1H,ArH),7.41(d,J＝5.1Hz,1H,ArH),7.36-7.31(m,5H,ArH),5.43-5.38(m,1H,HOCH₂ CH),5.18-5.14(m,1H,HOCH₂),4.73(d,J＝18.2Hz,1H,CONCH ₂),4.42(d,J＝18.4Hz,1H,CONCH ₂),4.06-3.97(m,2H,HO ₂CHCH),3.83(s,3H,N ₃CH).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):168.28,163.53,160.10,159.90,144.85,138.55,135.38,133.30,130.54,130.45,129.18,128.13,127.73,124.67,123.43,121.48,121.25,107.44,61.30,57.17,47.42,39.17.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₃N₆O₂:427.1882；Found:427.1871.

IR(cm^-1):3409.61,3267.17,2920.35,1677.93,1627.82,1574.07 1493.34,1450.57,1435.03,1365.18,1335.15,1299.97,1201.55,805.63,763.49,707.73,605.10.

Example 24

Synthesis of (S) -2- (2-hydroxy-1-phenylethyl) -6- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-24)

Synthesis of (S) -2- (2- ((tert-butyldimethylsilyl) oxy) -1-phenylethyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (X-2)

Dissolving a compound VIII-7(400mg,0.98mmol) and imidazole (133mg,1.96mmol) in dichloromethane (4mL), adding TBSCl (295mg,1.96mmol), stirring at 35 ℃ for reaction for about 8h, after TLC (dichloromethane: methanol ═ 30: 1) detection of the reaction of the raw material VIII-7, washing with saturated sodium bicarbonate (4mL), washing with saturated sodium chloride solution (4mL), drying with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure, adding petroleum ether (4mL), pulping, filtering to obtain a white solid 480mg, the yield is 93.6%, and m.p.70.2-72.1 ℃.

¹H-NMR(300MHz,DMSO-d₆))δ(ppm):9.13(d,J＝5.4Hz,1H,ArH),8.58-8.52(m,3H,ArH),7.83(d,J＝7.92Hz,1H,ArH),7.38-7.30(m.5H,ArH),5.45-5.41(m,1H,HOCH₂ CH),4.69(d,J＝18.5Hz,1H,CONCH ₂),4.46(d,J＝18.4Hz,1H,CONCH ₂),4.26-4.14(m,2H,O ₂CH),3.49(s,3H,S ₃CH),0.73(S,9H,C ₃ ₃(CH)),0.01(d,J＝4.7Hz,6H,Si ₃ ₂(CH)).

Dissolving 1-methyl-1H-pyrazole-5-amine (IX-2) (74mg,0.76mmol) in anhydrous tetrahydrofuran (3mL), cooling to-78 ℃, adding 1N LiHMDS (0.76mL,0.76mmol), stirring for 30min, dissolving compound X-2(200mg,0.38mmol) in anhydrous tetrahydrofuran (3mL), slowly dropwise adding the reaction solution at-78 ℃, reacting at-78 ℃ for about 1H, detecting by TLC (dichloromethane: methanol ═ 20:1) that raw material X-2 is completely reacted, evaporating the solvent under reduced pressure, adding ethyl acetate (5mL) to dissolve, adjusting pH to 2-3 with HCl ethyl acetate solution, continuing stirring for about 3H, detecting by TLC (dichloromethane: methanol ═ 15: 1) that the intermediate is completely reacted, evaporating the solvent under reduced pressure, separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-30: 1), yield 55mg of the target compound hydrochloride as a golden yellow solid: 31.3 percent and m.p.202.3-204.1 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.59(brs,1H,HN),8.61-8.60(m,1H,ArH),8.50(s,1H,ArH),8.40(d,J＝7.8Hz,1H,ArH),7.80(d,J＝8.3Hz,1H,ArH),7.63-7.61(m,1H,ArH),7.42-7.34(m,6H,ArH),6.37-6.33(m,1H,ArH),5.48-5.43(m,1H,HOCH₂ CH),5.22-5.18(m,1H,HOCH₂),4.78(d,J＝18.3Hz,1H,CH ₂NCO),4.47(d,J＝18.4Hz,1H,CH ₂NCO),4.12-4.03(m,2H,HO ₂CHCH),3.77(s,3H,N ₃CH).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):166.86,162.33,159.45,158.79,143.74,137.19,136.63,135.53,131.99,129.17,127.85,126.81,126.40,123.34,120.21,115.02,107.79,99.19,59.93,55.85,46.09,34.44.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₃N₆O₂:427.1882；Found:427.1867.

IR(cm^-1):3396.69,3233.24,2923.20,1666.59,1558.09,1449.72,1407.88,1307.11,1269.59,1203.11,1062.83,1042.64,820.99,768.85,700.35,613.30.

Example 25

Synthesis of (S) -2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-25)

Synthesis of (S) -2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- (methylsulfonyl) pyrimidin-4-yl) isoindolin-1-one (VIII-8)

Dissolving compound VI (2,25g,5.83mmol), (S) -2-amino-2- (3-chlorophenyl) ethyl-1-ol (VII-5) (1g,5,83mmol) and triethylamine (1.6mL,11.65mmol) in acetonitrile (30mL), heating and refluxing for about 6h, until the raw materials react completely by TLC (dichloromethane: methanol ═ 20:1), evaporating the solvent under reduced pressure, adding methanol (10mL), cooling in ice bath to below 10 ℃, stirring for 30min, precipitating a solid, and performing suction filtration to obtain 1.72g of a white solid, wherein the yield is: 66.5 percent and m.p.71.8-73.5 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.16-9.14(m,1H,ArH),8.60-8.55(m,3H,ArH),7.84(d,J＝8.8Hz,1H,ArH),7.44-7.34(m,4H,ArH),5.41-5.38(m,1H,CHCH₂OH),5.23-5.21(m,1H,CHCH₂ OH),4.78(d,J＝19.1Hz,1H,CH ₂N),4.51(d,J＝18.9Hz,1H,CH ₂N),4.08-4.00(m,2H,HO ₂CHCH),3.51(s,3H,S ₃CH).

Dissolving compound VIII-8(400mg,0.90mmol) and 4-aminotetrahydropyran (IX-1) (182mg,1.80mmol) in sec-butanol (3mL), heating to 125 ℃ with a sealed tube, reacting for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-8 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain 220mg of a light yellow solid, yield: 52.6 percent and m.p.86.2-87.9 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):8.44(s,1H,ArH),8.40-8.39(m,1H,ArH),8.34(d,J＝7.9Hz,1H,ArH),7.73(d,J＝7.9Hz,1H,ArH),7.45(s,1H,ArH),7.38-7.30(m,3H,ArH),7.26-7.25(m,1H,ArH),5.42-5.38(m,1H,HOCH₂ CH),5.24-5.23(m,1H,HOCH₂CH),4.74(d,J＝18.3Hz,1H,CONCH ₂),4.46(d,J＝18.0Hz,1H,CONCH ₂),4.05-4.03(m,3H,HO ₂CH,NHCH),3.92-3.89(m,2H,O(C ₂H) ₂),3.46-3.43(m,2H,O(C ₂H) ₂),1.92-1.88(m,2H,NHCH( ₂CH) ₂),1.62-1.51(m,2H,NHCH( ₂CH) ₂).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):168.30,161.98,159.72,150.22,144.68,141.21,137.42,133.74,132.98,130.99,130.51,128.04,127.61,126.51,124.52,121.45,106.05,66.57,61.07,56.84,47.66,47.10,32.82.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₂₆ClN₄O₃:465.1693；Found:465.1681.

IR(cm^-1):3324.61,2952.51,2843.82,1674.86,1573.21,1524.39,1492.66,1454.00,1410.36,1367.09,1248.13,1200.68,1138.06,1085.54,1011.43,979.63,868.32,807.30,770.97,613.89.

Example 26

Synthesis of (S) -2- (1- (3-chlorophenyl) -2-hydroxyethyl) -6- (2- ((1-methyl-1H-pyrazol-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-26)

Dissolving compound VIII-8(400mg,0.90mmol) and 1-methyl-1H-pyrazol-4-amine (IX-3) (175mg,1.80mmol) as raw materials in sec-butyl alcohol (3mL), heating to 125 ℃ in a sealed tube to react for about 12 hours, detecting that the raw material VIII-8 completely reacts by TLC (dichloromethane: methanol: 25:1), cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol: 200: 1-50: 1) to obtain a golden yellow solid 126mg, wherein the yield: 30.4 percent and m.p.207.8-209.6 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.58(s,1H,NH),8.53-8.51(m,1H,ArH),8.47(s,1H,ArH),8.39(d,J＝8.4Hz,1H,ArH),7.92(s,1H,ArH),7.77(d,J＝8.0Hz,1H,ArH),7.58(brs,1H,ArH),7.44-7.32(m,5H,ArH),5.41-5.37(m,1H,HOCH₂ CH),5.24-5.20(m,1H,HOCH₂CH),4.74(d,J＝17.9Hz,1H,CONCH ₂),4.48(d,J＝18.0Hz,1H,CONCH ₂),4.07-4.00(m,2H,HO ₂CH),3.83(s,3H,N ₃CH).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):168.27,163.47,160.10,159.89,144.91,141.20,137.36,133.76,133.09,131.06,130.61,130.45,128.08,127.62,126.55,124.70,123.44,121.52,121.21,107.41,61.16,56.80,47.68,39.09.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₂ClN₆O₂:461.1493；Found:461.1482.

IR(cm^-1):3263.50,3064.40,1673.56,1626.77,1594.58,1573.68,1455.10,1434.10,1333.59,1300.77,1262.55,1201.99,1076.02,1040.00,803.48,768.99,691.89,616.86.

Example 27

Synthesis of 2- (3-chlorobenzyl) -6- (2- ((4-fluorophenyl) amino) pyrimidin-4-yl) isoindolin-1-one (I-27)

Dissolving a compound VIII-1(200mg,0.48mmol) and para-fluoroaniline IX-13(81mg,0.73mmol) in sec-butyl alcohol (5mL), sealing a tube, heating to 125 ℃ for reaction for about 12h, detecting by TLC (dichloromethane: methanol ═ 25:1) that the reaction of the raw material VIII-1 is complete, cooling the reaction solution to room temperature, evaporating the solvent under reduced pressure, and separating by column chromatography (eluent: dichloromethane: methanol ═ 200: 1-50: 1) to obtain a gray solid 98mg, wherein the yield is as follows: 45.9 percent and 196.2 to 197.3 ℃.

¹H-NMR(300MHz,DMSO-d₆)δ(ppm):9.76(s,1H,NH),8.58(d,J＝5.2Hz,1H,ArH),8.52(s,1H,ArH),8.40(dd,J＝8.1,1.5Hz,1H,ArH),7.83(dd,J＝8.8,4.8Hz,2H,ArH),7.74(d,J＝8.0Hz,1H,ArH),7.54(d,J＝5.2Hz,1H,ArH),7.19-7.13(m,3H,ArH),7.28(d,J＝6.4Hz,1H,ArH),7.16(t,J＝8.9Hz,2H,ArH),4.78(s,2H,CH2),4.50(s,2H,CH₂).

¹³C-NMR(75MHz,DMSO-d₆)δ(ppm):167.05,162.88,161.41,159.71(d,J＝66.2Hz),159.22,144.36,139.97,136.79(d,J＝8.5Hz),136.71,132.72,130.60,130.02,128.80,127.54,127.37,126.38,124.18,121.32,120.62(d,J＝7.5Hz),114.96(d,J＝22.1Hz),108.11,49.53,44.96.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₅H₁₉ClFN₄O:445.1226；Found:445.1235.

IR(cm^-1):3465.54,3265.05,1703.97,1570.27,1537.36,1507.27,1463.32,1453.48,1430.72,1303.10,1199.33,829.97,801.87,764.94,711.27,612.21,548.43.

Example 28

Synthesis of 2- ((6-methylpyridin-2-yl) methyl) -6- (2- ((tetrahydro-2H-pyran-4-yl) amino) pyrimidin-4-yl) isoindolin-1-one (I-28)

Synthesis of methyl 2- (bromomethyl) -5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzoate (XVII)

Compound III (1g,3.6mmol) and NBS (765mg,4.3mmol) were dissolved in carbon tetrachloride (15mL), benzoyl peroxide (12mg,0.036mmol) was added, heated to 80 ℃ for about 4h, and starting material III was detected by TLC (petroleum ether: ethyl acetate ═ 20:1) for completion of the reaction. Cooling to room temperature, filtering off insoluble substances, concentrating the filtrate to obtain red oily substance, and directly feeding into the next step without treatment.

Synthesis of 2- ((6-methylpyridin-2-yl) methyl) -6- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) isoindolin-1-one (XVIII)

6-methyl-2-aminomethylpyridine (VII-5) (194mg,1.8mmol) and triethylamine (182mg,1.8mmol) were dissolved in acetonitrile (10mL), and compound XVII (714mg,2mmol) was added dropwise, followed by heating to 80 ℃ to react for about 10 hours. TLC (dichloromethane: methanol 20:1) detects complete reaction of the XVII starting material, the solvent is evaporated to dryness under reduced pressure, ethyl acetate (15mL) is dissolved, washed with water, dried, filtered, the filtrate is evaporated under reduced pressure to remove the solvent, and the residue is separated by column chromatography (eluent: dichloromethane: methanol 75: 1) to give 157mg of a dark green oil, yield: 23.9 percent.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.40(s,1H,ArH),7.99(d,J＝7.6Hz,1H ArH),7.57(t,J＝7.7Hz,1H ArH),7.45(d,J＝7.6Hz,1H ArH),7.14(d,J＝7.8Hz,1H ArH),7.10(d,J＝7.6Hz,1H ArH),4.97(s,2H,CH₂),4.49(s,2H,CH₂),2.60(s,3H,CH₃),1.39(s,12H,CH₃).

Synthesis of 6- (2-chloropyrimidin-4-yl) -2- ((6-methylpyridin-2-yl) methyl) isoindolin-1-one (XIX)

XVIII (500mg,1.4mmol), 2, 4-dichloropyrimidine (XX) (207mg,1.4mmol), palladium tetratriphenylphosphine (162mg,0.14mmol) and potassium phosphate (890mg,4.2mmol) were dissolved in a dioxane/water mixed solution (20/5mL), and the mixture was heated to 80 ℃ under nitrogen atmosphere to react for about 12 hours. TLC (dichloromethane: methanol 25:1) detects complete reaction of the starting material XVIII, cools to room temperature, filters, concentrates the filtrate under reduced pressure, and separates the residue by column chromatography (eluent: dichloromethane: methanol 50:1) to give 272mg of a yellow solid, yield: 55.4 percent and m.p.208.3-209.2 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.74-8.72(m,1H,ArH),8.54(s,1H,ArH),8.47(d,J＝7.9Hz,1H,ArH),7.82-7.74(m,1H,ArH),7.65-7.57(m,2H,ArH),7.17(d,J＝7.1Hz,1H,ArH),7.12(d,J＝6.8Hz,1H,ArH),4.97(s,2H,CH₂),4.60(s,2H,CH₂),2.60(s,3H,CH₃).

Mixing XIX (50mg,0.14mmol), 4-aminotetrahydropyran (IX-1) (14mg,0.14mmol), Pd (OAc)₂(3mg,0.014mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (35mg,0.06mmol) and Cs₂CO₃Dissolving (68mg,0.21mmol) in dioxane (3mL), heating to 100 ℃ under the protection of nitrogen, reacting for about 12 hours, detecting complete reaction of the raw material XIX by TLC (dichloromethane: methanol 20:1), cooling the reaction liquid to room temperature, adding ethyl acetate for dilution, washing with water (2mL) and saturated salt water (2mL), drying an organic layer with anhydrous sodium sulfate, filtering, evaporating the filtrate under reduced pressure to remove the solvent, and separating residue by column chromatography (eluent: dichloromethane: methanol 200: 1-50: 1) to obtain a light yellow solid 24mg, wherein the yield is as follows: 41.3 percent and m.p.186.1-187.2 ℃.

¹H-NMR(300MHz,CDCl₃)δ(ppm):8.48(s,1H,ArH),8.36(d,J＝5.3Hz,1H,ArH),8.27(d,J＝7.7Hz,1H,ArH),7.59-7.52(m,2H,ArH),7.15(d,J＝6.9Hz,1H,ArH),7.12-7.02(m,2H,ArH),5.32(s,1H,NH),4.95(s,2H,CH₂),4.55(s,2H,CH₂),4.29-4.10(m,1H,NHCH),4.04-4.00(m,2H,OCH₂),3.63-3.56(m,2H,OCH₂),2.58(s,3H,CH₃),2.13-2.08(m,2H,NHCHCH₂),1.67-1.57(m,2H,NHCHCH₂).

¹³C-NMR(75MHz,CDCl₃)δ(ppm):168.09,164.20,161.57,158.39,158.02,155.94,143.75,137.67,137.61,133.24,130.21,123.23,122.51,122.31,119.51,106.65,66.76,50.34,48.21,47.25,33.27,24.21.

HRMS(ESI):m/z[M+H]⁺.Calcd for C₂₄H₂₆N₅O₂:416.2087；Found:416.2081.

IR(cm^-1):3437.25,3260.22,2959.37,2920.76,2839.89,1570.28,1593.93,1529.95,1462.49,1412.85,1369.02,1136.31,1109.76,872.80,822.00,773.29,612.73,531.56.

Example 29

The pharmacological experiments and results of the partial compounds prepared in the above are as follows:

first, ERK2 enzyme inhibition activity experiment

(I) an experimental method

All compounds were dissolved in DMSO to prepare 10mM stock, and then the compounds were added to the screening system at concentrations ranging from 0.05nM to 1. mu.M for detection, and diluted in 3-fold gradients, each concentration being in duplicate wells. The experimental results are converted into the activity percentage, dose-effect curves are drawn, and inhibition IC is calculated by GRAPHPAD PEISM 5 nonlinear regression₅₀The value is obtained.

The enzyme reaction system consists of: 20mM Hepes (pH 7.5),10mM MgCl₂,1mM EGTA,0.02％Brij35,0.02mg/mL BSA,0.1mM Na₃VO₄2mM DTT,10 μ M ATP, kinase substrate; and simultaneously adding compounds to be screened with different concentrations to form a 50 mu l reaction system, reacting at room temperature for 2h, detecting the ADP content in the system by using a luciferase method, reacting for 5min, and detecting a chemiluminescence signal on an MD-SpectraMax M5 multifunctional enzyme-labeled instrument, wherein the value intensity of the chemiluminescence signal is in direct proportion to the inhibition of enzyme activity. Substituting the detected chemiluminescence signal value into the formula:

percentage of enzyme activity (%) { (Lu drug-Lu background)/(Lu enzyme-Lu background) } x 100%

IC of the compound was then calculated using Graphpad Prism5 software₅₀The value is obtained.

Test results:

partial compounds of the invention were screened for ERK2 kinase inhibitory activity in vitro and the results are shown in table 1.

TABLE 1 IC of part of the tested compounds for ERK2 kinase₅₀Value of

The results in Table 1 show that the compounds of the invention have better inhibitory activity on ERK2 kinase, wherein the compounds I-8 and I-10 have better activity and IC of ERK2 kinase₅₀0.7nM and 1.2nM, respectively. Meanwhile, the pharmacological activities of the S configuration and the R configuration of the compounds I-8 and I-10 are respectively compared, and the activity of the compound with the S configuration is far better than that of the compound with the R configuration.

Second, experiment of inhibition of human tumor cell proliferation

(I) an experimental method

Test grouping and drug concentration selection

All compound concentrations were selected: 1nM,10nM,100nM,500nM,1000 nM.

Cell Culture conditions (Cell Culture)

The cells were subcultured for 10-15 passages under conditions of medium containing penicillin (final concentration of 100U/mL), streptomycin (final concentration of 100. mu.g/mL), 10% FBS (CT26, Colo-205, WM-266-4: RPMI-1640; HCT116, HT 29: McCoy's 5 a; SW 626: L-15), when the cells fused to 90%, the old medium was discarded, the cells were washed with 2mL of PBS for 2 times, after discarding PBS, 2mL of 0.25% (w/v) Trypsin-0.53mM EDTA mixed digest was added, and when the cells were gently rounded, 2mL of complete medium was rapidly added to terminate the digestion, and the cells were collected by pipetting. Centrifuging at 800rpm and 4 deg.C for 5min, discarding supernatant, suspending cells with complete culture medium, culturing in bottles, and changing the culture medium every other day.

MTT assay

Cells in logarithmic growth phase were seeded at 1X 105 cells/well in 96-well plates, placed at 37 ℃ and 5% CO₂After culturing under conditions until the cells are 90% confluent, the cells are synchronized by incubating for 2h with serum-free L-15 medium. Subsequently, the supernatant was discarded, and each of the compounds (final dilution concentration 1nM,10nM,100nM,500nM,1000nM after compound addition) in L-15 medium was added for 72h, and 20. mu.l of MTT solution (5mg/mL) was added to each well 4h before the end of incubation. After incubation, the supernatant from each well was discarded, 150. mu.L of DMSO was added to each well, and the cells were shaken on a cell shakerAnd (5) after 10min, fully dissolving the crystal, and measuring OD570 by using an enzyme labeling instrument.

The percentage (%) of proliferation inhibitory activity was (OD value administration-OD value background)/(OD value control-OD value background) x 100%

The concentration of the drug was plotted on the abscissa and the percentage of the proliferation inhibitory activity corresponding to each concentration was plotted on the ordinate, and the IC of each compound was calculated by nonlinear regression using Graphpd Prism5₅₀The value is obtained.

Test results:

selecting a part of compounds with better ERK2 kinase inhibition activity, and determining the antiproliferative capability of the compounds on tumor cell lines containing RAS-RAF-MEK-ERK mutation, namely human colon cancer cell Colo-205(BRAFV600E), human melanoma cell WM-266-4(BRAFV600D), human ovarian cancer cell SW-626(KRASG12V) and human colon cancer cell HCT-116(KRASG 13D). The experimental results show (Table 2) that the tested compound has certain proliferation inhibition capacity on four cell lines, and the inhibition activity is equivalent to that of the positive drug BVD-523.

TABLE 2 antiproliferative effect of partial compounds on four ERK pathway-activating cell lines

On this basis, the proliferation inhibitory ability of the above compounds on human chronic myelogenous leukemia cell K562, a tumor cell line without RAS-RAF-MEK-ERK mutation, was determined (Table 3). The results of the experiments show that the test compound and BVD-523 have inhibitory activity (IC) on K562 cell proliferation₅₀All are more than 1 mu M) is obviously weaker than the 4 tumor cell lines containing RAS-RAF-MEK-ERK mutation, the antiproliferative activity is greatly weakened, and the tested compound has certain selectivity on kinase and can selectively inhibit ERK kinase.

TABLE 3 antiproliferative effect of partial compounds on ERK pathway inactive cell lines

Third, NCM-460 cytotoxicity assay

(I) an experimental method

Test grouping and drug concentration selection

All compound concentrations were selected: 10nM,100nM,500nM,1000nM,10000 nM.

Cell Culture conditions (Cell Culture)

Subculturing the cells under the condition of RPMI-1640 medium containing penicillin (final concentration of 100U/mL), streptomycin (final concentration of 100. mu.g/mL) and 10% FBS, discarding the old medium when the cells are fused to 90%, washing the cells with 2mL of PBS for 2 times, discarding the PBS, adding 2mL of 0.25% (w/v) Trypsin-0.53mM EDTA mixed digestive juice, observing under a microscope for about 30s, rapidly adding 2mL of complete medium to stop digestion after the cells are rounded, gently blowing and beating, and collecting the cells. Centrifuging at 800rpm and 4 deg.C for 5min, discarding supernatant, suspending cells with complete culture medium, culturing in bottles, and changing the culture medium every other day.

MTT assay

Cells in logarithmic growth phase were grown at 1X 10⁵cells/well were seeded in 96-well plates at 37 ℃ with 5% CO₂After culturing under conditions until the cells are 90% confluent, the cells are synchronized by incubating for 2h with serum-free L-15 medium. Subsequently, the supernatant was discarded, and each of them was incubated for 72 hours in RPMI-1640 medium containing each compound (10nM,100nM,1000nM,5000nM,10000nM) for 4 hours before the end of the incubation, and 20. mu.L of MTT solution (5mg/mL) was added to each well. After incubation is finished, discarding supernatant of each well, adding 150 mu L DMSO into each well, oscillating on a cell oscillator for 10min, and measuring OD (optical density) by using an enzyme-labeling instrument after crystals are fully dissolved₅₇₀。

Test results:

anti-proliferative Effect of some Compounds of Table 4 on NCM-460 cells

We selected compounds I-8 and I-10 and tested their antiproliferative potential on normal cells using the NCM-460 cell line (Table 4). The results show that two compounds grow on NCM-460 cellsAll showed very weak proliferation inhibition, IC₅₀The values were all > 10000 nM.

From the above data we found that compounds I-8 and I-10 were not significantly toxic to normal cell growth of NCM-460.

Fourth, HCT-116 nude mouse transplanted tumor inhibition experiment

(I) an experimental method

Grouping conditions of experimental animals:

negative control group (model group): 10 patients were administered intraperitoneally with physiological saline containing 5% DMSO;

group I-8: 10, the administration dose is 50 mg/kg;

BVD-523 group: 10, the administration dose is 50 mg/kg;

the administration mode comprises the following steps: the preparation is administered by intragastric administration once a day.

Establishment of a transplantation tumor model and drug treatment:

HCT116 cells are subcultured for 10-15 generations under the condition of 1640 medium containing penicillin (final concentration of 100U/mL), streptomycin (final concentration of 100. mu.g/mL) and 10% FBS, when the cells are fused to 90%, the old medium is discarded, the cells are washed with 2mL of PBS for 2 times, 2mL of 0.25% trypsin-0.02% EDTA mixed digestive solution is added after the PBS is discarded, the mixture is observed under a microscope for about 30s, when the cells are rounded, 2mL of complete medium is rapidly added to stop digestion, the cells are lightly blown and collected. Centrifuging at 800rpm and 4 deg.C for 5min, discarding supernatant, suspending cells with complete culture medium, culturing in bottles, and changing the culture medium every other day.

Taking HCT116 cells in logarithmic growth phase, preparing into 5 × 10 under aseptic condition with serum-free culture medium⁶Selecting 30 healthy nude mice from the above experiment, feeding for one week to reach a weight of 20g, injecting 0.2mL of cell suspension into left armpit of each nude mouse, with cell concentration of 5 × 10⁷and/mL. Measuring the diameter of the transplanted tumor of the nude mouse by using a vernier caliper until the tumor grows to 100mm³Animals were randomized after left and right as day 0 and then started dosing according to the dosing method in the group. The antitumor effect of the test compound was dynamically observed using a method of measuring the tumor diameter. The number of measurements of the tumor diameter isEvery 3 days. The administration volume was different for each group. After 30 days, the mice were sacrificed and the tumor mass was surgically removed and weighed. The formula for Tumor Volume (TV) is:

TV＝1/2×a×b²

wherein a and b represent length and width, respectively.

Statistical analysis:

statistical differences between the data groups were determined to be significant using one-way ANOVA and Tukey's tests, with P values less than 0.05.

Test results:

we selected compound I-8, evaluated the in vivo anti-tumor activity of compound I-8 using a mouse transplantation tumor model of HCT-116 tumor cells, and compared it to the positive drug BVD-523.

As shown in figure 1, after the compound I-8 and the BVD-523 are continuously administered by gavage for 30 days at 50mg/kg/day, the increase of the volume and the weight of the transplanted tumor of the HCT-116 nude mice can be remarkably inhibited (p is less than 0.05), the tumor inhibition rate of the I-8 is 71 percent, the tumor inhibition rate of the BVD-523 is 54 percent, and the both do not influence the body weight of the mice. Experimental results show that the anti-tumor effect of the I-8 is better than that of a positive drug BVD-523 under the condition of intragastric administration of 50 mg/kg.

Claims

1. An isoindolinone derivative represented by the general formula I or a pharmaceutically acceptable salt thereof:

wherein:

R¹is selected from

Wherein X is selected from CH₂O or CH-OH; r⁴Selected from H or C1-C6 alkyl, R⁵Selected from H, CH₃、-CH₂OH、-CH₂CH₂OH、-CH₂CH₂CH₂OH or-CH₂CH₂CH₂CH₂OH; y is selected from O, NH or S; r⁶Selected from F, Cl, Br, CH₃、NH₂Or NHCOCH₃；

R²Is H or-CH₂OH；

R³Is selected from

Wherein R is⁷Is H, F, Cl, Br or OCH₃，R⁷Is mono-, di-or tri-substituted.

2. Isoindolinone derivative I or a pharmaceutically acceptable salt thereof according to claim 1, wherein:

R¹is selected from

3. Isoindolinone derivative I or a pharmaceutically acceptable salt thereof according to claim 1, wherein:

R³is selected from

4. Isoindolinone derivative I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein: r²is-CH₂OH, the compound I or a pharmaceutically acceptable salt thereof includes chiral isomers:

5. isoindolinone derivative I according to claim 1, or a pharmaceutically acceptable salt thereof, which is selected from I-1 to I-26:

。

6. isoindolinone derivative I or a pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt is an acid addition salt of the compound of the general formula I, wherein the acid used for salt formation is: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.

7. A process for the preparation of isoindolinone derivatives according to claim 1, wherein compound I is prepared by substitution of compound VIII with compound IX:

8. a process for the preparation of isoindolinone derivatives according to claim 1, comprising the steps of:

reacting a compound VIII with tert-butyldimethylsilyl chloride TBSCl to prepare a compound X; then preparing a compound XI from the compound X and IX through substitution reaction; finally, compound XI is subjected to hydroxyl protecting group removal and salification to prepare compound I^.A。

9. A pharmaceutical composition comprising a compound of formula I of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

10. Use of the isoindolinone derivative according to claim 1 or a pharmaceutically acceptable salt thereof for the preparation of an ERK kinase inhibitor medicament.