WO2020107987A1 - Erk inhibitor containing isoindoline, preparation method therefor and application thereof - Google Patents

Erk inhibitor containing isoindoline, preparation method therefor and application thereof Download PDF

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WO2020107987A1
WO2020107987A1 PCT/CN2019/104110 CN2019104110W WO2020107987A1 WO 2020107987 A1 WO2020107987 A1 WO 2020107987A1 CN 2019104110 W CN2019104110 W CN 2019104110W WO 2020107987 A1 WO2020107987 A1 WO 2020107987A1
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compound
acid
reaction
methanol
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徐云根
纪德重
张灵芝
朱启华
柏英
吴尧尧
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中国药科大学
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

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  • the present invention belongs to medicinal chemistry technology, and specifically relates to a class of ERK kinase inhibitors containing 6-(2-aminopyrimidin-4-yl)isoindolin-1-one structure, their preparation methods, and medicines containing these compounds Composition and its use in the preparation of anti-tumor drugs.
  • the mitogen-activated protein kinases (MAPKs) signal transduction pathway is an important signal pathway that transduces cell surface signals to the nucleus, which causes cell proliferation by affecting the transcription and regulation of genes in animal cells , Differentiation, transformation and apoptosis.
  • MAPKs signaling pathways have been found in mammalian cells, including extracellular signal-regulated kinases (ERK, Extracellular Regulated Proteins) and c-Jun N-terminal kinases (JNK/SAPK, c-Jun N-terminal Kinase/stress- activated protein), p38 kinase isozymes (p38A, p38B, p38C and p38D), ERK3/ERK4 and ERK5.
  • ERK extracellular signal-regulated kinases
  • JNK/SAPK c-Jun N-terminal kinases
  • JNK/SAPK c-Jun N-terminal Kinase/stress- activated protein
  • p38 kinase isozymes p38A, p38B, p38C and p38D
  • ERK3/ERK4 ERK5
  • the RAS-RAF-MEK-ERK signal transduction pathway (ERK pathway) is an evolutionarily conserved signaling cascade reaction pathway that can be transferred to cell surface receptor signals, thereby promoting cell proliferation and survival. Under normal physiological conditions, the ERK signaling pathway plays an important role in maintaining cell stability and regulating cell growth, and is strictly controlled by a multi-level feedback regulatory pathway.
  • BRAF inhibitors and MEK inhibitors have achieved great success in the field of anti-tumor, but with the development of clinical applications, it is found that whether BRAF inhibitors are used alone or in combination with MEK inhibitors, most The patient's condition will worsen again within one year (acquired resistance). In addition, about 10% to 15% of tumor patients with B-Raf V600E mutations are insensitive to BRAF inhibitors and MEK inhibitors (intrinsic resistance). Therefore, the problem of drug resistance of BRAF and MEK inhibitors has become a key scientific problem that needs to be solved at present.
  • ERK inhibitors will first block the re-activation of the MAPKs pathway exhibited by tumor cells resistant to BRAF and MEK inhibitors, overcoming existing drug resistance problems. Secondly, tumors inevitably develop drug resistance mutations through long-term targeted drug therapy. As mentioned above, a large number of BRAF and MEK mutations are mentioned. But so far, almost no mutations in ERK1/2 have been observed in tumor cells. At the same time, preclinical experimental results of ERK inhibitors show better effects than BRAF and MEK inhibitors. Therefore, inhibiting ERK is more effective than blocking its upstream kinase to block the signal transduction of the ERK pathway and overcome the resistance of tumor cells to BRAF inhibitors and MEK inhibitors.
  • the present invention provides an isoindolinone derivative containing 6-(2-aminopyrimidin-4-yl)isoindolin-1-one structure, and provides specific preparation of the derivative Method and pharmaceutical application for preparing ERK kinase inhibitor.
  • the present invention discloses an isoindolinone derivative represented by general formula I or a pharmaceutically acceptable salt thereof:
  • R 1 is selected from Wherein, X is selected from CH 2 , O, NH or CH-OH; R 4 is selected from H or C1-C6 alkyl, R 5 is selected from H, CH 3 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH; Y is selected from CH 2 , O, NH or S; R 6 is selected from F, Cl, Br, CH 3 , NH 2 or NHCOCH 3 ;
  • R 2 is H or -CH 2 OH
  • R 3 is selected from Wherein, R 7 is H, F, Cl, Br or OCH 3 , R 7 is mono-substituted, di-substituted or tri-substituted; R 8 is H or CH 3 .
  • R 1 is selected from
  • R 3 is selected from
  • R 2 is -CH 2 OH
  • the compound I or a pharmaceutically acceptable salt thereof includes chiral isomers:
  • the isoindolinone derivatives described in this application are selected from I-1 to I-28:
  • the above pharmaceutically acceptable salts are acid addition salts of compounds of general formula I, wherein the acids used for salt formation are: 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 acids used for salt formation are: 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 compound of the general formula (I) of the present invention can be prepared by the following method:
  • the compound IX is 1-methyl-1H-pyrazole-4-amine, 1-methyl-1H-pyrazole-3-amine, 4-aminotetrahydropyran, 4-aminocyclohexyl Alcohol, 4-aminopiperidine, 3-aminopyrrolidine, ethanolamine, cyclohexylamine, 3-amino-1-propanol, isopropylamine, 4-aminopyridine;
  • the solvent used is selected from sec-butanol, tert-butanol, ethanol , Tetrahydrofuran, ethyl acetate, methanol or a mixed solvent of any two or three of them, preferably sec-butanol;
  • the reaction temperature is selected from 90°C to 150°C, preferably 110°C to 130°C;
  • the reaction time is 4h to 48h, The reaction time is preferably 8h to 24h.
  • the compound VIII: compound IX (molar ratio) ratio is 1:1 to 1:20, preferably 1:1.5 to 1:3.
  • the second preparation method of the compound of general formula (I) includes the following steps:
  • R 3 stands for: Compound I
  • a configuration comprises:. S-configuration, R configuration and a mixture of two racemic configuration.
  • the process of preparing compound X from the reaction of compound VIII with tert-butyldimethylchlorosilane (TBSCl) the base used is selected from triethylamine, 4-dimethylaminopyridine, imidazole, sodium carbonate or potassium carbonate, preferably imidazole; reaction solvent Preferably ethyl acetate, tetrahydrofuran, chloroform, acetonitrile, methylene chloride, toluene, or a mixed solvent of any two or three of them, preferably methylene chloride; the temperature should be selected from 0°C to 50°C, preferably 30°C to 40°C ; The reaction time is selected from 1h to 12h, preferably 7h to 9h; Compound VIII: tert-butyldimethylchlorosilane (TBSCl) (molar ratio) The ratio is selected from 1:1 to 1:10, preferably 1:1.5 to 1 :2.
  • the process of preparing compound XI from compound X and IX through substitution reaction said 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 bis (Trimethylsilyl) lithium amide;
  • the solvent used is tetrahydrofuran, N,N-dimethylformamide, dioxane, N-methylpyrrolidone or a mixed solvent of any two or three of them, preferably tetrahydrofuran And N,N-dimethylformamide;
  • the reaction temperature is selected from -80°C to 0°C, preferably -80°C to -25°C;
  • the reaction time is selected from 0.1h to 10h, preferably 1h to 3h;
  • the process of preparing compound I. A by removing the hydroxyl protecting group from compound XI to form a salt .
  • the deprotection reagent (A) used is selected from hydrogen chloride, hydrogen bromide, sulfuric acid or trifluoroacetic acid; the solvents are methanol, ethanol and methylene chloride , Acetone, ethyl acetate, tetrahydrofuran or a mixed solvent of any two; the reaction temperature is selected from 0°C to 50°C, preferably 0°C to 25°C; the reaction time is selected from 0.1h to 6h, preferably 1h to 3h.
  • the preparation method 1 of the key intermediate VIII in the preparation method of the general formula (I) of the present invention includes:
  • R 2 represents: H or -CH 2 OH
  • R 3 represents: Wherein R 7 represents H, F, Cl, Br or OCH 3 , R 7 may be mono-substituted, di-substituted or tri-substituted; R 8 represents H or CH 3 .
  • R 2 CH 2 OH
  • the configuration of compound VIII includes: S configuration, R configuration, and a mixed racemate of two configurations.
  • the catalyst used is selected from tetratriphenylphosphine palladium (Pd(PPh 3 ) 4 ), [1,1'-bis(diphenylphosphino ) Ferrocene) palladium dichloride (Pd(dppf)Cl 2 ), bistriphenylphosphine palladium dichloride (Pd(PPh3) 2 Cl 2 ), palladium acetate (Pd(OAc) 2 ) or (1, 1'-bis(diphenylphosphinoferrocene) nickel dichloride (NiCl 2 (dppf)), preferably tetratriphenylphosphine palladium (Pd(PPh 3 ) 4 ).
  • the base is selected from sodium ethoxide, sodium acetate, potassium acetate, potassium phosphate, sodium carbonate or potassium carbonate, preferably sodium carbonate;
  • the reaction solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, ethyl acetate Glycol dimethyl ether, 1,4-dioxane, tetrahydrofuran, toluene, ethanol, water or a mixed solvent of any two or three of them, preferably a mixed solvent of toluene/methanol/water;
  • the reaction temperature is selected from 60°C ⁇ 120°C, preferably 70°C ⁇ 80°C;
  • reaction time is selected from 4h ⁇ 24h, preferably 7h ⁇ 10h;
  • compound II: compound III: catalyst: base (molar ratio) ratio is selected from 1:1:0.02:1 ⁇ 6 :1:0.2:6, preferably 1:1:0.03:1 ⁇ 1:1.05:0.1:2.
  • Oxidation of compound IV to prepare compound V The oxidant used is selected from m-CPBA, hydrogen peroxide, sodium hypochlorite, periodate, sodium periodate, potassium permanganate or tert-peroxide Butanol, preferably m-chloroperoxybenzoic acid; the reaction solvent is selected from tetrahydrofuran, acetone, methylene chloride, 1,4-dioxane or acetonitrile, preferably methylene chloride; the reaction temperature is selected from 10°C to 80°C, preferably 20°C to 30°C; reaction time is selected from 0.5h to 5h, preferably 1h to 2h; compound IV: oxidant (molar ratio) ratio is selected from 1:1 to 1:10, preferably 1:1.5 to 1:3.
  • the brominated reagent 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°C to 100°C, preferably 70°C to 90°C;
  • the reaction time is selected from 1h to 18h, preferably 4h ⁇ 7h;
  • Compound V: brominated reagent (molar ratio) is selected from 1:1 to 1:20, preferably 1:1 to 1:2.
  • the base used is selected from triethylamine, benzotriazol-1-yl-oxytripyrrolidinylphosphonium hexafluorophosphate (PyBop), N, N-diisopropylethylamine (DIEA), sodium carbonate or potassium carbonate, preferably triethylamine
  • the reaction solvent is selected from tetrahydrofuran, acetone, acetonitrile, methanol, methylene chloride, or a mixed solvent of any two, preferably acetonitrile
  • reaction The temperature is selected from 50°C to 100°C, preferably 60°C to 90°C;
  • the reaction time is selected from 1h to 24h, preferably 5h to 9h;
  • the compound VI: compound VII: base (molar ratio) ratio is selected from 1:1: 1 ⁇ 1:4:8, preferably 1:1:1 to 1:1.5:1.5.
  • the preparation method 2 of the key intermediate VIII in the preparation method of the general formula (I) of the present invention includes:
  • R 2 represents: H
  • R 3 represents: Wherein R 7 represents H, F, Cl, Br or OCH 3 , R 7 may be mono-substituted, di-substituted or tri-substituted; R 8 represents H or CH 3 .
  • the reaction is carried out under the conditions of adding a catalyst, a base and a reaction solvent, wherein the catalyst is selected from bis(triphenylphosphine) palladium dichloride, tetrakis(III) Phenylphosphine) palladium or [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, preferably tetrakis(triphenylphosphine)palladium; base selected from sodium ethoxide, sodium acetate, potassium acetate , Potassium phosphate, sodium carbonate or potassium carbonate, preferably sodium carbonate; the reaction solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, ethylene glycol dimethyl ether, 1,4-bis Oxyhexane, tetrahydrofuran, toluene, methanol, ethanol, water, or
  • the deprotection agent used is 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°C to 50°C, preferably 20°C to 30°C;
  • the reaction time is selected from 0.5h to 6h, preferably 1h to 2h.
  • the process of preparing compound XVI by substitution reaction of compound XIV and compound XV the base used 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, ethylene glycol dimethyl ether, or a mixed solvent of any three, preferably 1,4-dioxane; the reaction temperature is selected from 0°C to 120°C, preferably 50 °C ⁇ 80 °C; reaction time is selected from 3h to 24h, preferably 9h to 12h; compound XIV: compound XV: base (molar ratio) ratio is selected from 1:1:1 to 1:6:6, preferably 1:1 :1 ⁇ 1:1.2:1.5.
  • Oxidation reaction of compound XVI to prepare compound VIII The oxidant used is selected from m-CPBA, hydrogen peroxide, sodium hypochlorite, periodate, sodium periodate, potassium permanganate or tertiary peroxide Butanol, preferably m-chloroperoxybenzoic acid; the reaction solvent is selected from tetrahydrofuran, acetone, methylene chloride, 1,4-dioxane or acetonitrile, preferably methylene chloride; the reaction temperature is selected from 10°C to 80°C, preferably 20°C to 30°C; the reaction time is selected from 0.1h to 10h, preferably 1h to 2h; the compound IV: oxidant (molar ratio) ratio is selected from 1:1 to 1:6, preferably 1:1.5 to 1:3.
  • the invention also discloses a pharmaceutical composition, which contains the compound of the general formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
  • the compound can be added to a pharmaceutically acceptable carrier to make common pharmaceutical preparations, such as tablets, capsules, syrups, suspensions, flavors, sweeteners, liquid or solid fillers or diluents can be added for common pharmaceutical use Accessories.
  • ERK kinase inhibitors are used to prepare drugs for treating malignant tumors.
  • the present invention discloses a new isoindolinone derivative represented by the general formula (I). Pharmacological experiments show that the compound I of the present invention can produce a good inhibitory effect on ERK kinase and can be used for preparation A medicine for treating malignant tumors with excessive activation of ERK pathway; the invention also discloses a preparation method of the isoindolinone derivatives.
  • Figure 1 is the experimental results of compound I-8 HCT116 nude mice xenografts.
  • the composition of the enzyme reaction system is as follows: 20 mM Hepes (pH 7.5), 10 mM MgCl 2 , 1 mM EGTA, 0.02% Brij35, 0.02 mg/mL BSA, 0.1 mM Na 3 VO 4 , 2 mM DTT, 10 ⁇ M ATP, kinase, kinase substrate; Add different concentrations of the compound to be screened to form a 50 ⁇ l reaction system, react at room temperature for 2h, then use the luciferase method to detect the ADP content in the system, and then react for 5min, on the MD-SpectraMax M5 multi-functional microplate reader The chemiluminescence signal is detected, and the intensity of the chemiluminescence signal is proportional to the inhibition of enzyme activity.
  • the detected chemiluminescence signal value is substituted into the formula:
  • Enzyme activity percentage (%) ⁇ (Lu drug-Lu background)/(Lu enzyme-Lu background) ⁇ x100%
  • Cells were subcultured for 10-15 generations, and the culture conditions were medium containing penicillin (final concentration 100U/mL), streptomycin (final concentration 100 ⁇ g/mL), 10% FBS (CT26, Colo-205, WM-266 -4: RPMI-1640; HCT116, HT29: McCoy's 5a; SW626: L-15), when the cells are fused to 90%, discard the old medium, wash the cells twice with 2mL PBS, add 2mL after discarding the PBS 0.25% (w/v) Trypsin-0.53 mM EDTA mixed digestion solution, observed under a microscope, about 30s, when the cells become round, quickly add 2mL of complete medium to stop the digestion, gently pipet to collect the cells. Centrifuge at 800 rpm, 4°C for 5 min, discard the supernatant, resuspend the cells in complete medium, culture in separate flasks, and change the medium the next day.
  • Log growth phase cells were seeded in 96-well plates at 1 ⁇ 105cells/well and cultured at 37°C under 5% CO 2 until the cells were 90% confluent, and incubated with serum-free L-15 medium for 2h Cell synchronization. Subsequently, the supernatant was discarded, and L-15 medium containing each compound (final dilution concentration of 1 nM, 10 nM, 100 nM, 500 nM, 1000 nM after compound addition) was added for incubation for 72 h. 4 h before the end of incubation, 20 ⁇ l of MTT solution was added to each well (5mg/mL).
  • Percentage of proliferation inhibitory activity (OD value administration-OD value background) / (OD value control-OD value background) x 100%
  • Some compounds with better ERK2 kinase inhibitory activity were selected and tested for human colon cancer cell line Colo-205 (BRAFV600E) and human melanoma cell WM-266-4 (BRAFV600D) containing RAS-RAF-MEK-ERK mutation ), anti-proliferative ability of human ovarian cancer cell SW-626 (KRASG12V) and human colon cancer cell HCT-116 (KRASG13D).
  • the experimental results showed (Table 2) that the test compounds showed a certain degree of proliferation inhibitory ability on the four cell lines, and the inhibitory activity was comparable to that of the positive drug BVD-523.
  • Subculture of cells the culture conditions are RPMI-1640 medium containing penicillin (final concentration 100U/mL), streptomycin (final concentration 100 ⁇ g/mL), 10% FBS, when the cells are confluent to 90%, discard Wash the cells twice with 2mL of PBS, discard the PBS, add 2mL of 0.25% (w/v) Trypsin-0.53mM EDTA mixed digestion solution, and observe under the microscope for about 30s. When the cells become round, add them quickly 2mL complete medium was used to stop the digestion, gently pipetting to collect the cells. Centrifuge at 800 rpm, 4°C for 5 min, discard the supernatant, resuspend the cells in complete medium, culture in separate flasks, and change the medium the next day.
  • Log growth phase cells were seeded in 96-well plates at 1 ⁇ 10 5 cells/well and cultured at 37°C under 5% CO 2 until the cells were 90% confluent and incubated with serum-free L-15 medium Synchronize the cells for 2h. Subsequently, the supernatant was discarded, and the RPMI-1640 medium containing each compound (10 nM, 100 nM, 1000 nM, 5000 nM, 10000 nM) was added for incubation for 72 h, and 4 ⁇ L of MTT solution (5 mg/mL) was added to each well before incubation for 4 h.
  • Negative control group 10 rats, intraperitoneal administration of saline containing 5% DMSO;
  • Group I-8 10 animals, the dosage is 50mg/kg;
  • BVD-523 group 10 animals, the dosage is 50mg/kg;
  • Mode of administration once a day, intragastric administration.
  • HCT116 cells were subcultured for 10-15 passages under 1640 medium containing penicillin (final concentration 100U/mL), streptomycin (final concentration 100 ⁇ g/mL), 10% FBS, when the cells were confluent to 90% , Discard the old culture medium, wash the cells twice with 2mL of PBS, add 2ml of 0.25% trypsin-0.02% EDTA mixed digestion solution after discarding the PBS, observe under the microscope, about 30s, when the cells become round, quickly add 2ml completely The medium is terminated for digestion, gently pipetting to collect the cells. Centrifuge at 800 rpm, 4°C for 5 min, discard the supernatant, resuspend the cells in complete medium, culture in separate flasks, and change the fluid the next day.
  • the statistical difference between the data groups was determined by one-way ANOVA and Tukey’s test. A P value of less than 0.05 was considered to be a significant difference.
  • compound I-8 and BVD-523 were able to significantly inhibit the increase in the volume and weight of transplanted tumors of HCT-116 nude mice (p ⁇ 0.05) after continuous oral administration at 50 mg/kg/day for 30 days.
  • the tumor suppression rate of I-8 is 71%, and the tumor suppression rate of BVD-523 is 54%, neither of which will affect the body weight of mice.
  • the experimental results show that the anti-tumor effect of I-8 at the dose of 50 mg/kg administered by gavage is superior to the positive drug BVD-523.

Abstract

An isoindoline derivative, in particular a compound containing a structure of 6-(2-aminopyrimidin-4-yl)isoindolinon-1-one; and a preparation method for the isoindoline derivative. Further disclosed are an application of the isoindoline derivative and a stereoisomer, hydrate, solvate or crystal thereof, as well as a composition comprising the isoindoline derivative or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable carrier in the preparation of ERK kinase inhibitors, and a use of a pharmaceutical composition containing such compounds in the preparation of anti-tumor drugs.

Description

含异吲哚啉酮的ERK抑制剂及其制备方法与用途ERK inhibitor containing isoindolinone, preparation method and application thereof 技术领域Technical field
本发明属于药物化学技术，具体涉及一类含有6-(2-氨基嘧啶-4-基)异吲哚啉-1-酮结构的ERK激酶抑制剂，它们的制备方法，以及含有这些化合物的药物组合物及其在制备抗肿瘤药物方面的用途。The present invention belongs to medicinal chemistry technology, and specifically relates to a class of ERK kinase inhibitors containing 6-(2-aminopyrimidin-4-yl)isoindolin-1-one structure, their preparation methods, and medicines containing these compounds Composition and its use in the preparation of anti-tumor drugs.
技术背景technical background
丝裂原活化蛋白激酶(Mitogen-activated protein kinases，MAPKs)信号转导通路是将细胞表面信号转导至细胞核的重要信号通路，该通路通过影响动物细胞内基因的转录和调控，从而引起细胞增殖、分化、转化以及凋亡等生物学反应。目前在哺乳动物细胞中共发现了5条MAPKs信号通路，包括细胞外信号调节激酶(ERK,Extracellular regulated protein kinases)、c-Jun N末端激酶(JNK/SAPK,c-Jun N-terminal kinase/stress-activated protein kinase)、p38激酶同工酶(p38A、p38B、p38C和p38D)、ERK3/ERK4以及ERK5。The mitogen-activated protein kinases (MAPKs) signal transduction pathway is an important signal pathway that transduces cell surface signals to the nucleus, which causes cell proliferation by affecting the transcription and regulation of genes in animal cells , Differentiation, transformation and apoptosis. Five MAPKs signaling pathways have been found in mammalian cells, including extracellular signal-regulated kinases (ERK, Extracellular Regulated Proteins) and c-Jun N-terminal kinases (JNK/SAPK, c-Jun N-terminal Kinase/stress- activated protein), p38 kinase isozymes (p38A, p38B, p38C and p38D), ERK3/ERK4 and ERK5.
RAS-RAF-MEK-ERK信号转导通路(ERK通路)是一个进化保守的信号级联反应通路，可以转到细胞表面受体信号，从而促进细胞增殖及存活。正常生理条件下ERK信号通路对于保持细胞稳定以及调节细胞生长具有重要作用，并且被多级反馈调节通路严格控制。The RAS-RAF-MEK-ERK signal transduction pathway (ERK pathway) is an evolutionarily conserved signaling cascade reaction pathway that can be transferred to cell surface receptor signals, thereby promoting cell proliferation and survival. Under normal physiological conditions, the ERK signaling pathway plays an important role in maintaining cell stability and regulating cell growth, and is strictly controlled by a multi-level feedback regulatory pathway.
到目前为止，BRAF抑制剂以及MEK抑制剂在抗肿瘤领域取得了较大成功，但是随着临床应用的开展，发现无论是单独使用BRAF抑制剂，或者BRAF抑制剂与MEK抑制剂联合用药，多数患者在一年以内病情会再次恶化(获得性耐药)。除此以外，约10％～15％携带B-Raf V600E突变的肿瘤患者对BRAF抑制剂以及MEK抑制剂不敏感(固有耐药)。因此，BRAF和MEK抑制剂耐药性的问题，成为目前需要解决的关键科学问题。 So far, BRAF inhibitors and MEK inhibitors have achieved great success in the field of anti-tumor, but with the development of clinical applications, it is found that whether BRAF inhibitors are used alone or in combination with MEK inhibitors, most The patient's condition will worsen again within one year (acquired resistance). In addition, about 10% to 15% of tumor patients with B-Raf V600E mutations are insensitive to BRAF inhibitors and MEK inhibitors (intrinsic resistance). Therefore, the problem of drug resistance of BRAF and MEK inhibitors has become a key scientific problem that needs to be solved at present.
与BRAF和MEK抑制剂相比，ERK抑制剂首先会再次阻断由对BRAF和MEK抑制剂耐药的肿瘤细胞所表现出的MAPKs通路的再次活化，克服现有的耐药性问题。其次，长时间通过靶向激酶的药物治疗，肿瘤不可避免的会产生耐药性突变，如上文提到大量BRAF和MEK突变。但是到目前为止，肿瘤细胞中几乎没有观察到ERK1/2的突变。同时，ERK抑制剂的临床前实验结果显示出相比于BRAF和MEK抑制剂更好的效果。因此，抑制ERK相比于抑制其上游激酶更能有效的阻断ERK通路的信号传导，克服肿瘤细胞对BRAF抑制剂以及MEK抑制剂的耐药性。Compared with BRAF and MEK inhibitors, ERK inhibitors will first block the re-activation of the MAPKs pathway exhibited by tumor cells resistant to BRAF and MEK inhibitors, overcoming existing drug resistance problems. Secondly, tumors inevitably develop drug resistance mutations through long-term targeted drug therapy. As mentioned above, a large number of BRAF and MEK mutations are mentioned. But so far, almost no mutations in ERK1/2 have been observed in tumor cells. At the same time, preclinical experimental results of ERK inhibitors show better effects than BRAF and MEK inhibitors. Therefore, inhibiting ERK is more effective than blocking its upstream kinase to block the signal transduction of the ERK pathway and overcome the resistance of tumor cells to BRAF inhibitors and MEK inhibitors.
发明内容Summary of the invention
发明目的：本发明提供了一种含有6-(2-氨基嘧啶-4-基)异吲哚啉-1-酮结构的异吲哚啉酮类衍生物，并提供了该衍生物的具体制备方法和应用于制备ERK激酶抑制剂的制药应用。Object of the invention: The present invention provides an isoindolinone derivative containing 6-(2-aminopyrimidin-4-yl)isoindolin-1-one structure, and provides specific preparation of the derivative Method and pharmaceutical application for preparing ERK kinase inhibitor.
技术方案：本发明公开了一种如通式I所示的异吲哚啉酮类衍生物或其药学上可接受的盐：Technical Solution: The present invention discloses an isoindolinone derivative represented by general formula I or a pharmaceutically acceptable salt thereof:
Figure PCTCN2019104110-appb-000001
Figure PCTCN2019104110-appb-000001
其中：among them:
R 1选自
Figure PCTCN2019104110-appb-000002
Figure PCTCN2019104110-appb-000003
其中，X选自CH 2、O、NH或CH-OH；R 4选自H或C1～C6的烷基，R 5选自H、CH 3、-CH 2OH、-CH 2CH 2OH、-CH 2CH 2CH 2OH或-CH 2CH 2CH 2CH 2OH；Y选自CH 2、O、NH或S；R 6选自F、Cl、Br、CH 3、NH 2或NHCOCH 3； R 1 is selected from
Figure PCTCN2019104110-appb-000002
Figure PCTCN2019104110-appb-000003
Wherein, X is selected from CH 2 , O, NH or CH-OH; R 4 is selected from H or C1-C6 alkyl, R 5 is selected from H, CH 3 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH; Y is selected from CH 2 , O, NH or S; R 6 is selected from F, Cl, Br, CH 3 , NH 2 or NHCOCH 3 ;
R 2为H或-CH 2OH； R 2 is H or -CH 2 OH;
R 3选自
Figure PCTCN2019104110-appb-000004
其中，R 7为H、F、Cl、Br或OCH 3，R 7为单取代、双取代或三取代；R 8为H或CH 3。 R 3 is selected from
Figure PCTCN2019104110-appb-000004
Wherein, R 7 is H, F, Cl, Br or OCH 3 , R 7 is mono-substituted, di-substituted or tri-substituted; R 8 is H or CH 3 .
作为优选，R 1选自
Figure PCTCN2019104110-appb-000005
Figure PCTCN2019104110-appb-000006
Preferably, R 1 is selected from
Figure PCTCN2019104110-appb-000005
Figure PCTCN2019104110-appb-000006
作为优选，R 3选自
Figure PCTCN2019104110-appb-000007
Preferably, R 3 is selected from
Figure PCTCN2019104110-appb-000007
需要说明的是，当R 2为-CH 2OH时，所述化合物I或其药学上可接受的盐包括手性异构体：
Figure PCTCN2019104110-appb-000008
It should be noted that when R 2 is -CH 2 OH, the compound I or a pharmaceutically acceptable salt thereof includes chiral isomers:
Figure PCTCN2019104110-appb-000008
优选的，本申请所述所述异吲哚啉酮类衍生物选自I-1至I-28：Preferably, the isoindolinone derivatives described in this application are selected from I-1 to I-28:
Figure PCTCN2019104110-appb-000009
Figure PCTCN2019104110-appb-000009
Figure PCTCN2019104110-appb-000010
Figure PCTCN2019104110-appb-000010
上述药学上可接受的盐为通式I化合物的酸加成盐，其中用于成盐的酸为：氯化氢、溴化氢、硫酸、碳酸、草酸、柠檬酸、琥珀酸、酒石酸、磷酸、乳酸、丙酮酸、乙酸、马来酸、甲磺酸、苯磺酸、对甲苯磺酸或阿魏酸。The above pharmaceutically acceptable salts are acid addition salts of compounds of general formula I, wherein the acids used for salt formation are: 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.
本发明通式(I)的化合物可用下列方法制备：The compound of the general formula (I) of the present invention can be prepared by the following method:
通式(I)化合物的制备方法一：Preparation method 1 of compound of general formula (I):
Figure PCTCN2019104110-appb-000011
Figure PCTCN2019104110-appb-000011
其中R 1代表：
Figure PCTCN2019104110-appb-000012
其中X代表CH 2、O、CH-OH；R 4代表H、C1～C6的烷基，R 5代表H、CH 3、-CH 2OH、-CH 2CH 2OH、-CH 2CH 2CH 2OH或-CH 2CH 2CH 2CH 2OH；R 2代表：H或-CH 2OH；R 3代表：
Figure PCTCN2019104110-appb-000013
当R 2＝CH 2OH时，化合物I构型包括：S构型、R构型及两构型的混合消旋体。 Where R 1 represents:
Figure PCTCN2019104110-appb-000012
Where X represents CH 2 , O, CH-OH; R 4 represents H, C1-C6 alkyl, R 5 represents H, CH 3 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH; R 2 represents: H or -CH 2 OH; R 3 represents:
Figure PCTCN2019104110-appb-000013
When R 2 =CH 2 OH, the configuration of Compound I includes: S configuration, R configuration and a mixed racemate of two configurations.
进一步的，所述的化合物IX为1-甲基-1H-吡唑-4-胺、1-甲基-1H-吡唑-3-胺、4-氨基四氢吡喃、4-氨基环己醇、4-氨基哌啶、3-氨基吡咯烷、乙醇胺、环己胺、3-氨基-1-丙醇、异丙胺、4-氨基吡啶；所用溶剂选自仲丁醇、叔丁醇、乙醇、四氢呋喃、乙酸乙酯、甲醇或其中任意两种或三种溶剂的混合溶剂，优选仲丁醇；反应温度选自 90℃～150℃，优选110℃～130℃；反应时间为4h～48h，反应时间优选8h～24h。所述化合物VIII：化合物IX(摩尔比)配比为1:1～1:20，优选1:1.5～1:3。Further, the compound IX is 1-methyl-1H-pyrazole-4-amine, 1-methyl-1H-pyrazole-3-amine, 4-aminotetrahydropyran, 4-aminocyclohexyl Alcohol, 4-aminopiperidine, 3-aminopyrrolidine, ethanolamine, cyclohexylamine, 3-amino-1-propanol, isopropylamine, 4-aminopyridine; the solvent used is selected from sec-butanol, tert-butanol, ethanol , Tetrahydrofuran, ethyl acetate, methanol or a mixed solvent of any two or three of them, preferably sec-butanol; the reaction temperature is selected from 90°C to 150°C, preferably 110°C to 130°C; the reaction time is 4h to 48h, The reaction time is preferably 8h to 24h. The compound VIII: compound IX (molar ratio) ratio is 1:1 to 1:20, preferably 1:1.5 to 1:3.
通式(I)化合物的制备方法二，包括以下步骤：The second preparation method of the compound of general formula (I) includes the following steps:
Figure PCTCN2019104110-appb-000014
Figure PCTCN2019104110-appb-000014
由化合物VIII与叔丁基二甲基氯硅烷TBSCl反应制备化合物X；再由化合物X与IX经取代反应制备化合物XI；最后由化合物XI脱去羟基保护基后成盐制备化合物I .A。 Then replaced by compounds IX and X was prepared by reaction of compound XI;; from the compound VIII with t-butyl dimethyl chlorosilane compound X TBSCI prepared from compound XI end of the reaction after removal of hydroxy protecting groups to the preparation of salts of Compound I A..
其中R 1代表：
Figure PCTCN2019104110-appb-000015
R 3代表：
Figure PCTCN2019104110-appb-000016
Figure PCTCN2019104110-appb-000017
化合物I .A构型包括：S构型、R构型及两构型的混合消旋体。 Where R 1 represents:
Figure PCTCN2019104110-appb-000015
R 3 stands for:
Figure PCTCN2019104110-appb-000016
Figure PCTCN2019104110-appb-000017
Compound I A configuration comprises:. S-configuration, R configuration and a mixture of two racemic configuration.
需要特别说明的是上述制备方法二也适用于R 2＝H。 It should be noted that the above preparation method 2 is also applicable to R 2 =H.
由化合物VIII与叔丁基二甲基氯硅烷(TBSCl)反应制备化合物X的过程：所用碱选自三乙胺、4-二甲氨基吡啶、咪唑、碳酸钠或碳酸钾，优选咪唑；反应溶剂优选乙酸乙酯、四氢呋喃、氯仿、乙腈、二氯甲烷、甲苯或其中任意两种或三种溶剂的混合溶剂，优选二氯甲烷；应温度选自0℃～50℃，优选30℃～40℃；反应时间选自1h～12h，优选7h～9h；化合物VIII：叔丁基二甲基氯硅烷(TBSCl)(摩尔比)配比选自1:1～1:10，优选1:1.5～1:2。The process of preparing compound X from the reaction of compound VIII with tert-butyldimethylchlorosilane (TBSCl): the base used is selected from triethylamine, 4-dimethylaminopyridine, imidazole, sodium carbonate or potassium carbonate, preferably imidazole; reaction solvent Preferably ethyl acetate, tetrahydrofuran, chloroform, acetonitrile, methylene chloride, toluene, or a mixed solvent of any two or three of them, preferably methylene chloride; the temperature should be selected from 0°C to 50°C, preferably 30°C to 40°C ; The reaction time is selected from 1h to 12h, preferably 7h to 9h; Compound VIII: tert-butyldimethylchlorosilane (TBSCl) (molar ratio) The ratio is selected from 1:1 to 1:10, preferably 1:1.5 to 1 :2.
由化合物X与IX经取代反应制备化合物XI的过程，所述的化合物IX为2-氨基噻唑、2-氨基噁唑、1-甲基-5-氨基四氮唑、1-甲基-1H-3-氨基吡唑或1-甲基-1H-5-氨基吡唑；所用碱选自双(三甲基硅基)氨基锂、双(三甲基硅基)氨基钠或氢化钠，优选双(三甲基硅基)氨基锂；所用溶剂为四氢呋喃、N,N-二甲基甲酰胺、二氧六环、N-甲基吡咯烷酮或其中任意两种或三种溶剂的混合溶剂，优选四氢呋喃和N,N-二甲基甲酰胺；反应温度选自-80℃～0℃，优选-80℃～-25℃；反应时间选自0.1h～10h，优选1h～3h；化合物X：化合物IX：碱(摩尔比)配比选自1:1:1～1:6:6，优选1:1.5:1.5～1:2:2。The process of preparing compound XI from compound X and IX through substitution reaction, said 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 bis (Trimethylsilyl) lithium amide; the solvent used is tetrahydrofuran, N,N-dimethylformamide, dioxane, N-methylpyrrolidone or a mixed solvent of any two or three of them, preferably tetrahydrofuran And N,N-dimethylformamide; the reaction temperature is selected from -80°C to 0°C, preferably -80°C to -25°C; the reaction time is selected from 0.1h to 10h, preferably 1h to 3h; Compound X: Compound IX : The ratio of base (molar ratio) is selected from 1:1:1 to 1:6:6, preferably 1:1.5:1.5 to 1:2:2.
由化合物XI脱去羟基保护基后成盐制备化合物I .A的过程，所用的脱保护试剂(A)选自氯化氢、溴化氢、硫酸或三氟乙酸；溶剂为甲醇、乙醇、二氯甲烷、丙酮、乙酸乙酯、四氢呋喃或任意两者的混合溶剂；反应温度选自0℃～50℃，优选0℃～25℃；反应时间选自0.1h～6h，优选1h～3h。 The process of preparing compound I. A by removing the hydroxyl protecting group from compound XI to form a salt . The deprotection reagent (A) used is selected from hydrogen chloride, hydrogen bromide, sulfuric acid or trifluoroacetic acid; the solvents are methanol, ethanol and methylene chloride , Acetone, ethyl acetate, tetrahydrofuran or a mixed solvent of any two; the reaction temperature is selected from 0°C to 50°C, preferably 0°C to 25°C; the reaction time is selected from 0.1h to 6h, preferably 1h to 3h.
本发明通式(I)制备方法中的关键中间体VIII的制备方法一，包括：The preparation method 1 of the key intermediate VIII in the preparation method of the general formula (I) of the present invention includes:
Figure PCTCN2019104110-appb-000018
Figure PCTCN2019104110-appb-000018
其中R 2代表：H或-CH 2OH；R 3代表：
Figure PCTCN2019104110-appb-000019
其中R 7代表H、F、Cl、Br或OCH 3，R 7可以是单取代、双取代或三取代；R 8代表H或CH 3。当R 2＝CH 2OH时，化合物VIII构型包括：S构型、R构型及两构型的混合消旋体。 Where R 2 represents: H or -CH 2 OH; R 3 represents:
Figure PCTCN2019104110-appb-000019
Wherein R 7 represents H, F, Cl, Br or OCH 3 , R 7 may be mono-substituted, di-substituted or tri-substituted; R 8 represents H or CH 3 . When R 2 =CH 2 OH, the configuration of compound VIII includes: S configuration, R configuration, and a mixed racemate of two configurations.
由化合物II和化合物III进行Suzuki偶联反应制备化合物IV的过程：所用的催化剂选自四三苯基膦钯(Pd(PPh 3) 4)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(Pd(dppf)Cl 2)、双三苯基磷二氯化钯(Pd(PPh3) 2Cl 2)、醋酸钯(Pd(OAc) 2)或(1,1'-双(二苯基膦)二茂铁)二氯化镍(NiCl 2(dppf))，优选四三苯基膦钯(Pd(PPh 3) 4)。碱选自乙醇钠、乙酸钠、乙酸钾、磷酸钾、碳酸钠或碳酸钾，优选碳酸钠；反应溶剂选自N,N-二甲基甲酰胺、N,N-二甲基乙酰胺、乙二醇二甲醚、1,4-二氧六环、四氢呋喃、甲苯、乙醇、水或其中任意两种或三种溶剂的混合溶剂，优选甲苯/甲醇/水混合溶剂；反应温度选自60℃～120℃，优选70℃～80℃；反应时间选自4h～24h，优选7h～10h；化合物II：化合物III：催化剂：碱(摩尔比)配比选自1:1:0.02:1～6:1:0.2:6，优选1:1:0.03:1～1:1.05:0.1:2。 Suzuki coupling reaction of compound II and compound III to prepare compound IV: the catalyst used is selected from tetratriphenylphosphine palladium (Pd(PPh 3 ) 4 ), [1,1'-bis(diphenylphosphino ) Ferrocene) palladium dichloride (Pd(dppf)Cl 2 ), bistriphenylphosphine palladium dichloride (Pd(PPh3) 2 Cl 2 ), palladium acetate (Pd(OAc) 2 ) or (1, 1'-bis(diphenylphosphinoferrocene) nickel dichloride (NiCl 2 (dppf)), preferably tetratriphenylphosphine palladium (Pd(PPh 3 ) 4 ). The base is selected from sodium ethoxide, sodium acetate, potassium acetate, potassium phosphate, sodium carbonate or potassium carbonate, preferably sodium carbonate; the reaction solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, ethyl acetate Glycol dimethyl ether, 1,4-dioxane, tetrahydrofuran, toluene, ethanol, water or a mixed solvent of any two or three of them, preferably a mixed solvent of toluene/methanol/water; the reaction temperature is selected from 60°C ～120℃, preferably 70℃～80℃; reaction time is selected from 4h～24h, preferably 7h～10h; compound II: compound III: catalyst: base (molar ratio) ratio is selected from 1:1:0.02:1～6 :1:0.2:6, preferably 1:1:0.03:1～1:1.05:0.1:2.
由化合物IV进行氧化反应制备化合物V的过程：所用的氧化剂选自间氯过氧苯甲酸(m-CPBA)、双氧水、次氯酸钠、高碘酸、高碘酸钠、高锰酸钾或过氧叔丁醇，优选间氯过氧苯甲酸；反应溶剂选自四氢呋喃、丙酮、二氯甲烷、1,4-二氧六环或乙腈，优选二氯甲烷；反应温度选自10℃～80℃，优选20℃～30℃；反应时间选自0.5h～5h，优选1h～2h；化合物IV：氧化剂(摩尔比)配比选自1:1～1:10，优选1:1.5～1:3。Oxidation of compound IV to prepare compound V: The oxidant used is selected from m-CPBA, hydrogen peroxide, sodium hypochlorite, periodate, sodium periodate, potassium permanganate or tert-peroxide Butanol, preferably m-chloroperoxybenzoic acid; the reaction solvent is selected from tetrahydrofuran, acetone, methylene chloride, 1,4-dioxane or acetonitrile, preferably methylene chloride; the reaction temperature is selected from 10°C to 80°C, preferably 20°C to 30°C; reaction time is selected from 0.5h to 5h, preferably 1h to 2h; compound IV: oxidant (molar ratio) ratio is selected from 1:1 to 1:10, preferably 1:1.5 to 1:3.
由化合物V经溴代反应制备化合物VI的过程：所用的溴代试剂选自N-溴代琥珀酰亚胺(NBS)或液溴，优选N-溴代琥珀酰亚胺；溶剂选自苯、四氯化碳、氯仿、乙腈、二氯甲烷、甲醇或甲苯，优选四氯化碳；反应温度选自50℃～100℃，优选70℃～90℃；反应时间选自1h～18h，优选4h～7h；化合物V：溴代试剂(摩尔比)配比选自1:1～1:20，优选1:1～1:2。Process for preparing compound VI from compound V through bromination reaction: the brominated reagent 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°C to 100°C, preferably 70°C to 90°C; the reaction time is selected from 1h to 18h, preferably 4h ~7h; Compound V: brominated reagent (molar ratio) is selected from 1:1 to 1:20, preferably 1:1 to 1:2.
由化合物VI与化合物VII经环合反应制备化合物VIII的过程：所用的碱选自三乙胺、六氟磷酸苯并***-1-基-氧基三吡咯烷基磷(PyBop)、N,N-二异丙基乙胺(DIEA)、碳酸钠或碳酸钾，优选三乙胺；反应溶剂选自四氢呋喃、丙酮、乙腈、甲醇、二氯甲烷或任意两者的混合溶剂，优选乙腈；反应温度选自50℃～100℃，优选60℃～90℃；反应时间选自1h～24h，优选5h～9h；化合物VI:化合物VII:碱(摩尔比)配比选自1:1:1～1:4:8，优选1:1:1～1:1.5:1.5。The process of preparing compound VIII from compound VI and compound VII by cyclization reaction: the base used is selected from triethylamine, benzotriazol-1-yl-oxytripyrrolidinylphosphonium hexafluorophosphate (PyBop), N, N-diisopropylethylamine (DIEA), sodium carbonate or potassium carbonate, preferably triethylamine; the reaction solvent is selected from tetrahydrofuran, acetone, acetonitrile, methanol, methylene chloride, or a mixed solvent of any two, preferably acetonitrile; reaction The temperature is selected from 50°C to 100°C, preferably 60°C to 90°C; the reaction time is selected from 1h to 24h, preferably 5h to 9h; the compound VI: compound VII: base (molar ratio) ratio is selected from 1:1: 1～ 1:4:8, preferably 1:1:1 to 1:1.5:1.5.
本发明通式(I)制备方法中的关键中间体VIII的制备方法二，包括：The preparation method 2 of the key intermediate VIII in the preparation method of the general formula (I) of the present invention includes:
Figure PCTCN2019104110-appb-000020
Figure PCTCN2019104110-appb-000020
其中R 2代表：H；R 3代表：
Figure PCTCN2019104110-appb-000021
其中R 7代表H、F、Cl、Br或OCH 3，R 7可以是单取代、双取代或三取代；R 8代表H或CH 3。 Where R 2 represents: H; R 3 represents:
Figure PCTCN2019104110-appb-000021
Wherein R 7 represents H, F, Cl, Br or OCH 3 , R 7 may be mono-substituted, di-substituted or tri-substituted; R 8 represents H or CH 3 .
由化合物II和化合物XII进行Suzuki偶联反应制备化合物XIII的过程：反应是在加入催化剂、碱和反应溶剂条件下进行，其中催化剂选自双(三苯基膦)二氯化钯、四(三苯基膦)钯或[1,1'-双(二苯基膦基)二茂铁]二氯化钯，优选四(三苯基膦)钯；碱选自乙醇钠、乙酸钠、乙酸钾、磷酸钾、碳酸钠或碳酸钾，优选碳酸钠；反应溶剂选自N,N-二甲基甲酰胺、N,N-二甲基乙酰胺、乙二醇二甲醚、1,4-二氧六环、四氢呋喃、甲苯、甲醇、乙醇、水或其中任意两种或三种溶剂的混合溶剂，优选甲苯/甲醇/水的混合溶剂；反应温度选自50℃～120℃，优选60℃～80℃；反应时间选自3h～30h，优选7h～12h；化合物II：化合物XII：催化剂：碱(摩尔比)配比选自1:1:0.02:1～6:1:0.3:10，优选1:1:0.05:1～1:1.05:0.1:2。Suzuki coupling reaction of compound II and compound XII to prepare compound XIII: the reaction is carried out under the conditions of adding a catalyst, a base and a reaction solvent, wherein the catalyst is selected from bis(triphenylphosphine) palladium dichloride, tetrakis(III) Phenylphosphine) palladium or [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, preferably tetrakis(triphenylphosphine)palladium; base selected from sodium ethoxide, sodium acetate, potassium acetate , Potassium phosphate, sodium carbonate or potassium carbonate, preferably sodium carbonate; the reaction solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, ethylene glycol dimethyl ether, 1,4-bis Oxyhexane, tetrahydrofuran, toluene, methanol, ethanol, water, or a mixed solvent of any two or three of them, preferably a mixed solvent of toluene/methanol/water; the reaction temperature is selected from 50°C to 120°C, preferably 60°C to 80℃; reaction time is selected from 3h to 30h, preferably 7h to 12h; compound II: compound XII: catalyst: base (molar ratio) ratio is selected from 1:1:0.02:1 to 6:1:0.3:10, preferably 1:1:0.05:1～1:1.05:0.1:2.
由化合物XIII经脱保护反应制备化合物XIV的过程，所用的脱保护剂选自氯化氢、三氟乙酸或三氟化硼，优选三氟乙酸；反应溶剂选自四氢呋喃、丙酮、二氯甲烷、1,4-二氧六环或乙腈，优选二氯甲烷；反应温度选自0℃～50℃，优选20℃～30℃；反应时间选自0.5h～6h，优选1h～2h。The process of preparing compound XIV from compound XIII through deprotection reaction, the deprotection agent used is 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°C to 50°C, preferably 20°C to 30°C; the reaction time is selected from 0.5h to 6h, preferably 1h to 2h.
由化合物XIV和化合物XV进行取代反应制备化合物XVI的过程：所用的碱选自碳酸钾、碳酸钠、磷酸钾、醋酸钾、氢氧化钾、氢化钠或醋酸钠，优选氢化钠；反应溶剂选自甲苯、甲醇、水、1,4-二氧六环、乙二醇二甲醚或任意三者的混合溶剂，优选1,4-二氧六环；反应温度选自0℃～120℃，优选50℃～80℃；反应时间选自3h～24h，优选9h～12h；化合物XIV:化合物XV:碱(摩尔比)配比选自1:1:1～1:6:6，优选1:1:1～1:1.2:1.5。The process of preparing compound XVI by substitution reaction of compound XIV and compound XV: the base used 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, ethylene glycol dimethyl ether, or a mixed solvent of any three, preferably 1,4-dioxane; the reaction temperature is selected from 0°C to 120°C, preferably 50 ℃ ~ 80 ℃; reaction time is selected from 3h to 24h, preferably 9h to 12h; compound XIV: compound XV: base (molar ratio) ratio is selected from 1:1:1 to 1:6:6, preferably 1:1 :1～1:1.2:1.5.
由化合物XVI进行氧化反应制备化合物VIII的过程：所用的氧化剂选自间氯过氧苯甲酸(m-CPBA)、双氧水、次氯酸钠、高碘酸、高碘酸钠、高锰酸钾或过氧叔丁醇，优选间氯过氧苯甲酸；反应溶剂选自四氢呋喃、丙酮、二氯甲烷、1,4-二氧六环或乙腈，优选二氯甲烷；反应温度选自10℃～80℃，优选20℃～30℃；反应时间选自0.1h～10h，优选1h～2h；化合物IV：氧化剂(摩尔比)配比选自1:1～1:6，优选1:1.5～1:3。Oxidation reaction of compound XVI to prepare compound VIII: The oxidant used is selected from m-CPBA, hydrogen peroxide, sodium hypochlorite, periodate, sodium periodate, potassium permanganate or tertiary peroxide Butanol, preferably m-chloroperoxybenzoic acid; the reaction solvent is selected from tetrahydrofuran, acetone, methylene chloride, 1,4-dioxane or acetonitrile, preferably methylene chloride; the reaction temperature is selected from 10°C to 80°C, preferably 20°C to 30°C; the reaction time is selected from 0.1h to 10h, preferably 1h to 2h; the compound IV: oxidant (molar ratio) ratio is selected from 1:1 to 1:6, preferably 1:1.5 to 1:3.
本发明还公开了一种药物组合物，其含有上述通式I化合物或其药学上可接受的盐及药学上可接受的载体。所述的化合物可以添加药学上可接受的载体制成常见的药用制剂，如片剂、胶囊、糖浆、悬浮剂，可以加入香料、甜味剂、液体或固体填料或稀释剂等常用药用辅料。The invention also discloses a pharmaceutical composition, which contains the compound of the general formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The compound can be added to a pharmaceutically acceptable carrier to make common pharmaceutical preparations, such as tablets, capsules, syrups, suspensions, flavors, sweeteners, liquid or solid fillers or diluents can be added for common pharmaceutical use Accessories.
本发明所述的通式I的异吲哚啉酮类衍生物及其立体异构体、水合物、溶剂合物或结晶在制备ERK激酶抑制剂药物中的应用也在本发明的保护范围内。The application of the isoindolinone derivatives of the general formula I and their stereoisomers, hydrates, solvates or crystals described in the present invention in the preparation of ERK kinase inhibitor drugs is also within the scope of the present invention .
进一步的，其中ERK激酶抑制剂用于制备治疗恶性肿瘤的药物。Further, ERK kinase inhibitors are used to prepare drugs for treating malignant tumors.
有益效果：本发明公开了一种新的通式(I)所示的异吲哚啉酮类衍生物，药理实验显示，本发明的化合物I可以对ERK激酶产生良好的抑制作用，可用于制备治疗ERK通路过度活化的恶性肿瘤的药物；本发明还公开了所述异吲哚啉酮类衍生物的制备方法。Beneficial effect: The present invention discloses a new isoindolinone derivative represented by the general formula (I). Pharmacological experiments show that the compound I of the present invention can produce a good inhibitory effect on ERK kinase and can be used for preparation A medicine for treating malignant tumors with excessive activation of ERK pathway; the invention also discloses a preparation method of the isoindolinone derivatives.
附图说明BRIEF DESCRIPTION
图1是化合物I-8的HCT116裸小鼠移植瘤实验结果。Figure 1 is the experimental results of compound I-8 HCT116 nude mice xenografts.
具体实施方式detailed description
下面结合具体实施例对本申请作出详细说明。The following describes the application in detail with reference to specific embodiments.
实施例1Example 1
2-(3-氯苄基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-1)的合成2-(3-chlorobenzyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindolin-1-one (I-1) Synthesis
6-(2-(甲硫基)嘧啶-4-基)-1-氧代异吲哚啉-2-甲酸叔丁酯(XIII)的合成Synthesis of 6-(2-(methylthio)pyrimidin-4-yl)-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (XIII)
将4-氯-2-(甲硫基)嘧啶(XII)(6.14g,38.24mmol)，6-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂-2-基)异吲哚啉-2-甲酸叔丁基酯(II)(13.74g,38.24mmol)，碳酸钠(8.11g,76.48mmol)和四三苯基膦钯(2.21g,1.91mmol)溶于由甲苯(150mL)、乙醇(50mL)和水(50mL)组成的混合溶剂中，氮气保护下，85℃反应12h左右，待TLC(二氯甲烷：甲醇＝35:1)检测反应完全，冷却至室温，减压蒸干溶剂，残留物加入乙酸乙酯(300mL)打浆，过滤，滤饼用乙酸乙酯洗涤后再经柱层析(洗脱剂：二氯甲烷：甲醇＝100：1)分离，得土黄色固体12.3g，产率90.0％，m.p.197.7～199.2℃。4-chloro-2-(methylthio)pyrimidine (XII) (6.14g, 38.24mmol), 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborola -2-yl)isoindoline-2-carboxylic acid tert-butyl ester (II) (13.74g, 38.24mmol), sodium carbonate (8.11g, 76.48mmol) and tetratriphenylphosphine palladium (2.21g, 1.91mmol) ) Dissolved in a mixed solvent composed of toluene (150mL), ethanol (50mL) and water (50mL), under nitrogen protection, react at 85 ℃ for about 12h, wait for TLC (dichloromethane: methanol = 35:1) to detect the completion of the reaction After cooling to room temperature, the solvent was evaporated to dryness under reduced pressure. The residue was added with ethyl acetate (300 mL) for slurry, filtered, and the filter cake was washed with ethyl acetate and then subjected to column chromatography (eluent: dichloromethane: methanol = 100: 1) Isolate to obtain 12.3g of earthy yellow solid, yield 90.0%, mp197.7～199.2℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),2.65(s,3H,SCH 3),1.61(s,9H,3×CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.60 (d, J=5.2 Hz, 1H, ArH), 8.53 (s, 1H, ArH), 8.47 (dd, J=8.0, 1.6 Hz, 1H , ArH), 7.61 (d, J = 8.0 Hz, 1H, ArH), 7.44 (d, J = 5.3 Hz, 1H, ArH), 4.84 (s, 2H, CH 2 ), 2.65 (s, 3H, SCH 3 ), 1.61(s, 9H, 3×CH 3 ).
6-(2-(甲硫基)嘧啶-4-基)异吲哚啉-1-酮(XIV)的合成Synthesis of 6-(2-(methylthio)pyrimidin-4-yl)isoindolin-1-one (XIV)
将化合物XIII(8.0g,22.4mmol)溶于二氯甲烷(80mL)，加入三氟乙酸(20mL)，室温搅拌1h左右，待TLC(二氯甲烷：甲醇＝25:1)检测原料反应完全，停止反应，减压蒸干溶剂后，残留物中加入水(100ml)，用饱和氢氧化钠溶液调节pH6～7，析出白色固体，过滤，烘干得到白色固体5.5g，产率：95.5％，m.p.>250.0℃。Compound XIII (8.0g, 22.4mmol) was dissolved in dichloromethane (80mL), trifluoroacetic acid (20mL) was added, stirred at room temperature for about 1h, until TLC (dichloromethane: methanol = 25:1) detected the raw material reaction was complete, After stopping the reaction and evaporating the solvent under reduced pressure, water (100 ml) was added to the residue, and the pH was adjusted to 6-7 with saturated sodium hydroxide solution. A white solid was precipitated, filtered, and dried to obtain 5.5 g of white solid. Yield: 95.5%. mp>250.0℃.
1H-NMR(300MHz,DMSO-d 6)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,CH 2),2.61(s,3H,CH 3). 1 H-NMR (300 MHz, DMSO-d 6 ) 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, CH 2 ), 2.61(s, 3H, CH 3 ).
2-(3-氯苄基)-6-(2-(甲硫基)嘧啶-4-基)异吲哚啉-1-酮(XVI-1)的合成Synthesis of 2-(3-chlorobenzyl)-6-(2-(methylthio)pyrimidin-4-yl)isoindolin-1-one (XVI-1)
将化合物XIV(2g,7.8mmol)溶于1,4-二氧六环(50mL)，加入60％氢化钠(376mg,9.4mmol)，室温搅拌1h，加入3-氯溴苄(2.4g,11.7mmol)，70℃搅拌反应12h左右，TLC(石油醚：乙酸乙酯＝2:1)检测原料XIV反应完全后，减压蒸干溶剂，经柱层析(洗脱剂：石油醚：乙酸乙酯＝6：1)分离，得黄色固体1.93g，产率：64.8％，m.p.152.2～153.4℃。Compound XIV (2g, 7.8mmol) was dissolved in 1,4-dioxane (50mL), 60% sodium hydride (376mg, 9.4mmol) was added, stirred at room temperature for 1h, and 3-chlorobromobenzyl (2.4g, 11.7) was added mmol), stirring at 70°C for about 12 hours, after TLC (petroleum ether: ethyl acetate=2:1) detects that the raw material XIV reaction is complete, the solvent is evaporated under reduced pressure and subjected to column chromatography (eluent: petroleum ether: ethyl acetate) Ester = 6:1) isolated to obtain 1.93 g of yellow solid, yield: 64.8%, mp 152.2 ~ 153.4 °C.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.37(s,2H,CH 2),2.66(s,3H,SCH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.61 (d, J=5.3 Hz, 1H, ArH), 8.55 (d, J=1.0 Hz, 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, CH 2 ), 4.37 (s, 2H, CH 2 ), 2.66 (s, 3H, SCH 3 ).
2-(3-氯苄基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-1)的合成Synthesis of 2-(3-chlorobenzyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII-1)
将化合物XVI-1(1.0g,2.6mmol)溶于二氯甲烷(20mL)，冰浴下加入间氯过氧苯甲酸(1.31g,6.5mmol)，室温搅拌2h左右，TLC(二氯甲烷：甲醇＝35:1)检测原料XVI-1反应完全，向反应液中加入饱和硫代硫酸钠溶液(30mL)，剧烈搅拌10min，静置分层，水层继续用二氯甲烷(30mLx3)萃取，合并有机层，用饱和碳酸钠溶液(30mLx3)洗涤，再用饱和氯化钠溶液洗涤(30mL)，无水硫酸钠干燥，过滤，减压蒸干溶剂，得白色固体890mg，产率：82.7％，m.p.145.3～146.1℃。Compound XVI-1 (1.0g, 2.6mmol) was dissolved in dichloromethane (20mL), m-chloroperoxybenzoic acid (1.31g, 6.5mmol) was added under ice bath, stirred at room temperature for about 2h, TLC (dichloromethane: Methanol=35:1) The raw material XVI-1 was detected to be completely reacted. Saturated sodium thiosulfate solution (30 mL) was added to the reaction solution, stirred vigorously for 10 min, and allowed to stand for separation. The aqueous layer was further extracted with methylene chloride (30 mL×3) The organic layers were combined, washed with saturated sodium carbonate solution (30 mL×3), and then with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to obtain a white solid 890 mg, yield: 82.7% , Mp145.3～146.1℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.41(s,2H,CH 2),3.46(s,3H,SO 2CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.99 (d, J=5.3 Hz, 1H, ArH), 8.59 (d, J=0.6 Hz, 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, CH 2 ), 4.41 (s, 2H, CH 2 ), 3.46 (s, 3H, SO 2 CH 3 ).
2-(3-氯苄基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-1)的合成2-(3-chlorobenzyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindolin-1-one (I-1) Synthesis
将化合物VIII-1(200mg,0.48mmol)和4-氨基四氢吡喃(IX-1)(74mg,0.73mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料反应完全，将反应液冷却至室温，减压蒸干溶剂，经柱层析(洗脱剂：二氯甲烷：甲醇＝200：1)分离，得黄色固体116mg，产率：55.6％，m.p.181.9～183.0℃。Compound VIII-1 (200mg, 0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg, 0.73mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol = 25:1) detects that the raw materials are completely reacted, the reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, and separated by column chromatography (eluent: dichloromethane: methanol = 200:1) , 116mg yellow solid was obtained, yield: 55.6%, mp181.9 ~ 183.0 ℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.35(s,2H,CH 2),4.26-4.10(m,1H,NHC H),4.04-4.00(m,2H,OC H 2 ),3.63-3.56(m,2H,OC H 2 ),2.12-2.08(m,2H,NHCHC H 2 ),1.66-1.56(m,2H,NHCHC H 2 ). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.49 (s, 1H, ArH), 8.37 (d, J=4.0 Hz, 1H, ArH), 8.27 (d, J=7.6 Hz, 1H, ArH ), 7.51 (d, J = 7.9 Hz, 1H, ArH), 7.35-7.25 (m, 3H, ArH), 7.21 (d, J = 3.4 Hz, 1H, ArH), 7.06 (d, J = 4.7 Hz, 1H, ArH), 5.37-5.22 (m, 1H, NH), 4.81 (s, 2H, CH 2 ), 4.35 (s, 2H, CH 2 ), 4.26-4.10 (m, 1H, NHC H ), 4.04- 4.00(m,2H,OC H 2 ),3.63-3.56(m,2H,OC H 2 ),2.12-2.08(m,2H,NHCHC H 2 ),1.66-1.56(m,2H,NHCHC H 2 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 24H 24ClN 4O 2:435.1582；Found:435.1590. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 24 ClN 4 O 2 : 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. 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.
实施例2Example 2
2-(3-氟-4-氯苄基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-2)的合成2-(3-fluoro-4-chlorobenzyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindolin-1-one ( I-2) Synthesis
2-(4-氯-3-氟苄基)-6-(2-(甲硫基)嘧啶-4-基)异吲哚啉-1-酮(XVI-2)的合成Synthesis of 2-(4-chloro-3-fluorobenzyl)-6-(2-(methylthio)pyrimidin-4-yl)isoindolin-1-one (XVI-2)
将化合物XIV(1g,3.9mmol)溶于1,4-二氧六环(50mL)，加入60％氢化钠(188mg,4.7mmol)，室温搅拌1h，加入4-氯-3-氟溴苄(1.3g,5.85mmol)，70℃搅拌12h左右，待TLC(石油醚：乙酸乙酯＝2:1)检测原料XIV反应完全，减压蒸干溶剂，残留物经柱层析(洗脱剂：石油醚：乙酸乙酯＝6：1)分离，得白色固体855mg，产率：54.8％，m.p.143.2～144.5℃。Compound XIV (1 g, 3.9 mmol) was dissolved in 1,4-dioxane (50 mL), 60% sodium hydride (188 mg, 4.7 mmol) was added, stirred at room temperature for 1 h, and 4-chloro-3-fluorobenzyl bromide was added ( 1.3g, 5.85mmol), stirred at 70℃ for about 12h, after TLC (petroleum ether: ethyl acetate=2:1) detects that the raw material XIV reaction is complete, the solvent is evaporated under reduced pressure, and the residue is subjected to column chromatography (eluent: Petroleum ether: ethyl acetate=6:1) separated to obtain 855 mg of white solid, yield: 54.8%, mp 143.2-144.5°C.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.40(s,2H,CH 2),2.70(s,3H,SCH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.63 (d, J=5.2 Hz, 1H, ArH), 8.57 (s, 1H, ArH), 8.43 (dd, J=8.0, 1.6 Hz, 1H , ArH), 7.58 (d, J = 8.0 Hz, 1H, ArH), 7.49 (d, J = 5.3 Hz, 1H, ArH), 7.41 (t, J = 7.8 Hz, 1H, ArH), 7.12 (ddd, J = 9.5, 8.9, 1.6 Hz, 2H, ArH), 4.83 (s, 2H, CH 2 ), 4.40 (s, 2H, CH 2 ), 2.70 (s, 3H, SCH 3 ).
2-(4-氯-3-氟苄基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-2)的合成Synthesis of 2-(4-chloro-3-fluorobenzyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII-2)
将化合物XVI-2(500mg,1.3mmol)溶于二氯甲烷(20mL)，冰浴下加入间氯过氧苯甲酸(657mg,3.25mmol)，室温搅拌2h左右，待TLC(二氯甲烷：甲醇＝35:1)检测原料XVI-2反应完全，向反应液中加入饱和硫代硫酸钠溶液(20mL)剧烈搅拌10min，静置分层，水层继续用二氯甲烷(20mLx3)萃取，合并有机层用饱和碳酸钠(20mLx3)洗涤，再用饱和氯化钠溶液洗涤(20mL)，无水硫酸钠干燥，过滤，减压蒸干溶剂，得白色固体447mg，产率：79.6％，m.p.153.8～155.2℃。Compound XVI-2 (500mg, 1.3mmol) was dissolved in dichloromethane (20mL), m-chloroperoxybenzoic acid (657mg, 3.25mmol) was added under ice bath, stirred at room temperature for about 2h, until TLC (dichloromethane: methanol) =35:1) The raw material XVI-2 was detected to be completely reacted. Saturated sodium thiosulfate solution (20 mL) was added to the reaction solution and stirred vigorously for 10 min. The mixture was allowed to stand for separation. The aqueous layer was further extracted with dichloromethane (20 mL×3). The layer was washed with saturated sodium carbonate (20mLx3) and then with saturated sodium chloride solution (20mL), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to obtain 447mg of white solid, yield: 79.6%, mp 153.8～ 155.2℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.41(s,2H,CH 2),3.45(s,3H,SO 2CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.99 (d, J=5.3 Hz, 1H, ArH), 8.58 (s, 1H, ArH), 8.53 (d, J=7.9 Hz, 1H, ArH ), 8.03 (d, J = 5.3 Hz, 1H, ArH), 7.62 (d, J = 8.0 Hz, 1H, ArH), 7.38 (t, J = 7.9 Hz, 1H, ArH), 7.13 (d, J = 9.1Hz, 1H, ArH), 7.07 (d, J = 8.2Hz, 1H, ArH), 4.81 (s, 2H, CH 2 ), 4.41 (s, 2H, CH 2 ), 3.45 (s, 3H, SO 2 CH 3 ).
2-(3-氟-4-氯苄基)-6-(2-((四氢化-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-2)的合成2-(3-fluoro-4-chlorobenzyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindolin-1-one ( I-2) Synthesis
将化合物VIII-2(207mg,0.48mmol)和4-氨基四氢吡喃(IX-1)(74mg,0.73mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料反应完全，将反应液冷却至室温，减压蒸干溶剂，经柱层析(洗脱剂：二氯甲烷：甲醇＝200：1)分离，得白色固体115mg，产率：52.9％，m.p.180.4～181.9℃。Compound VIII-2 (207mg, 0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg, 0.73mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol = 25:1) detects that the raw materials are completely reacted, the reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, and separated by column chromatography (eluent: dichloromethane: methanol = 200:1) , To obtain 115mg of white solid, yield: 52.9%, mp180.4 ~ 181.9 ℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.36(s,2H,C H 2 ),4.26-4.11(m,1H,NHC H),4.04-4.00(m,2H,OC H 2 ),3.63-3.56(m,2H,OC H 2 ),2.12-2.08(m,2H,NHCHC H 2 ),1.68-1.55(m,2H,NHCHC H 2 ). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.49 (s, 1H, ArH), 8.37 (d, J=3.8 Hz, 1H, ArH), 8.28 (dd, J=8.0, 1.5 Hz, 1H , ArH), 7.52 (d, J = 7.9 Hz, 1H, ArH), 7.37 (t, J = 7.8 Hz, 1H, ArH), 7.19-6.99 (m, 3H, ArH), 5.30 (brs, 1H, NH ), 4.80 (s, 2H, CH 2 ), 4.36 (s, 2H, CH 2 ), 4.26-4.11 (m, 1H, NHC H ), 4.04-4.00 (m, 2H, OC H 2 ), 3.63 3.56 (m, 2H, OC H 2 ), 2.12-2.08 (m, 2H, NHCHC H 2 ), 1.68-1.55 (m, 2H, NHCHC H 2 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) δ (ppm): 168.11, 164.03, 160.77 (d, J = 124.1 Hz), 158.52, 156.63, 143.19, 137.86 (d, J = 10.9 Hz), 133.16, 132.92, 131.04 , 130.45, 124.39 (d, J = 3.7 Hz), 123.29, 122.64, 116.26 (d, J = 21.3 Hz), 115.00, 106.65, 66.78, 49.50, 47.25, 45.66, 33.26.
HRMS(ESI):m/z[M+H] +.Calcd for C 24H 23ClFN 4O 2:453.1488；Found:453.1494. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 23 ClFN 4 O 2 : 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. IR (cm -1 ): 3247.73, 3482.25, 2292.60, 2843.90, 1701.76, 1602.04, 1570.09, 1528.32, 1414.52, 1369.96, 1162.42, 1196.23, 1055.28, 867.41, 802.31, 768.02, 612.67, 535.91.
实施例3Example 3
2-(3-氯-4-氟苄基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-3)的合成2-(3-chloro-4-fluorobenzyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindolin-1-one ( I-3) Synthesis
2-(3-氯-4-氟苄基)-6-(2-(甲硫基)嘧啶-4-基)异吲哚啉-1-酮(XVI-3)的合成Synthesis of 2-(3-chloro-4-fluorobenzyl)-6-(2-(methylthio)pyrimidin-4-yl)isoindolin-1-one (XVI-3)
将化合物XIV(1g,3.9mmol)溶于1,4-二氧六环(50mL)，加入60％氢化钠(188mg,4.7mmol)，室温搅拌1h，加入4-氟-3-氯溴苄(1.3g,5.85mmol)，70℃搅拌12h左右，待TLC(石油醚：乙酸乙酯＝2:1)检测原料XIV反应完全，减压蒸干溶剂，经柱层析(洗脱剂：石油醚：乙酸乙酯＝6：1)分离，得橙色固体933mg，产率：59.8％，m.p.174.9～176.3℃。Compound XIV (1 g, 3.9 mmol) was dissolved in 1,4-dioxane (50 mL), 60% sodium hydride (188 mg, 4.7 mmol) was added, stirred at room temperature for 1 h, and 4-fluoro-3-chlorobromobenzyl ( 1.3g, 5.85mmol), stir at 70℃ for about 12h, wait for TLC (petroleum ether: ethyl acetate = 2:1) to detect the completion of the raw material XIV reaction, evaporate the solvent under reduced pressure, and perform column chromatography (eluent: petroleum ether) : Ethyl acetate=6:1) Separation, to obtain 933mg of orange solid, yield: 59.8%, mp174.9 ~ 176.3 ℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.37(s,2H,CH 2),2.67(s,3H,SCH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.60 (d, J=5.0 Hz, 1H, ArH), 8.54 (s, 1H, ArH), 8.40 (d, J=8.0 Hz, 1H, ArH ), 7.55 (d, J = 7.9 Hz, 1H, ArH), 7.46 (d, J = 5.2 Hz, 1H, ArH), 7.38 (d, J = 6.8 Hz, 1H, ArH), 7.21 (brs, 1H, ArH), 7.12 (t, J = 8.5 Hz, 1H, ArH), 4.78 (s, 2H, CH 2 ), 4.37 (s, 2H, CH 2 ), 2.67 (s, 3H, SCH 3 ).
2-(3-氯-4-氟苄基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-3)的合成Synthesis of 2-(3-chloro-4-fluorobenzyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII-3)
将化合物XVI-3(500mg,1.25mmol)溶于二氯甲烷(20ml)，冰浴下加入间氯过氧苯甲酸(657mg,3.25mmol)，室温搅拌2h左右，待TLC(二氯甲烷：甲醇＝35:1)检测原料XVI-3反应完全，向反应液中加入饱和硫代硫酸钠溶液(20mL)，剧烈搅拌10min，静置分层，水层继续用二氯甲烷(20mLx3)萃取，合并有机层，用饱和碳酸钠(20mLx3)洗涤，再用饱和氯化钠溶液洗涤(20mL)，无水硫酸钠干燥，过滤，减压蒸干溶剂，得白色固体446mg，产率：82.6％，m.p.173.2～174.4℃。Compound XVI-3 (500mg, 1.25mmol) was dissolved in dichloromethane (20ml), m-chloroperoxybenzoic acid (657mg, 3.25mmol) was added under ice bath, stirred at room temperature for about 2h, until TLC (dichloromethane: methanol) =35:1) The raw material XVI-3 was tested for complete reaction. Saturated sodium thiosulfate solution (20 mL) was added to the reaction solution, stirred vigorously for 10 min, and allowed to stand for separation. The aqueous layer was further extracted with dichloromethane (20 mL x 3) and combined The organic layer was washed with saturated sodium carbonate (20mLx3) and then with saturated sodium chloride solution (20mL), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to give 446mg of white solid, yield: 82.6%, mp 173.2～174.4℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.40(s,2H,CH 2),3.46(s,3H,SO 2CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.99 (d, J=5.3 Hz, 1H, ArH), 8.57 (s, 1H, ArH), 8.53 (dd, J=8.0, 1.7 Hz, 1H , ArH), 8.02 (d, J = 5.3 Hz, 1H, ArH), 7.62 (d, J = 8.0 Hz, 1H, ArH), 7.39 (dd, J = 6.8, 2.1 Hz, 1H, ArH), 7.26 7.18 (m, 1H, ArH), 7.13 (t, J = 8.6 Hz, 1H, ArH), 4.80 (s, 2H, CH 2 ), 4.40 (s, 2H, CH 2 ), 3.46 (s, 3H, SO 2 CH 3 ).
2-(3-氯-4-氟苄基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-3)的合成2-(3-chloro-4-fluorobenzyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindolin-1-one ( I-3) Synthesis
将化合物VIII-3(207mg,0.48mmol)和4-氨基四氢吡喃(IX-1)(74mg,0.73mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-3反应完全，将反应液冷却至室温，减压蒸干溶剂，经柱层析(洗脱剂：二氯甲烷：甲醇＝200：1)分离，得白色固体104mg，产率：47.8％，m.p.210.5～211.7℃。Compound VIII-3 (207mg, 0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg, 0.73mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol = 25:1) detects that the raw material VIII-3 is completely reacted. The reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, and subjected to column chromatography (eluent: dichloromethane: methanol = 200: 1) Isolate to obtain 104 mg of white solid, yield: 47.8%, mp210.5-211.7°C.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.35(s,2H,CH 2),4.26-4.09(m,1H,NHCH),4.04-4.00(m,2H,OC H 2 ),3.63-3.56(m,2H,OC H 2 ),2.12-2.08(m,2H,NHCHC H 2 ),1.68-1.55(m,2H,NHCHC H 2 ). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.49 (s, 1H, ArH), 8.37 (d, J=5.2 Hz, 1H, ArH), 8.27 (dd, J=8.0, 1.4 Hz, 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, CH 2 ), 4.35 (s, 2H, CH 2 ), 4.26-4.09 (m, 1H, NHCH), 4.04-4.00 ( m, 2H, OC H 2 ), 3.63-3.56 (m, 2H, OC H 2 ), 2.12-2.08 (m, 2H, NHCHC H 2 ), 1.68-1.55 (m, 2H, NHCHC H 2 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) δ (ppm): 168.07, 164.00, 163.26, 160.49 (d, J = 171.3 Hz), 158.58, 143.19, 137.92, 134.01 (d, J = 3.6 Hz), 132.99, 130.40 , 130.29, 127.95, 127.86, 123.28, 122.62, 116.99 (d, J = 21.3 Hz), 106.65, 66.78, 49.45, 47.24, 45.49, 33.27.
HRMS(ESI):m/z[M+H] +.Calcd for C 24H 23ClFN 4O 2:453.1488；Found:453.1492. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 23 ClFN 4 O 2 : 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. IR (cm -1 ): 3461.03, 3249.92, 2942.39, 2847.62, 1701.05, 1604.79, 1571.85, 1531.09, 1493.95, 1261.83, 1238.03, 868.32, 803.01, 761.81, 628.16
实施例4Example 4
2-(3-氯苄基)-6-(2-((1-甲基-1H-吡唑-5-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-4)的合成2-(3-chlorobenzyl)-6-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)isoindolin-1-one (I- 4) Synthesis
将化合物VIII-1(200mg,0.48mmol)和1-甲基-1H-吡唑-5-胺(IX-2)(71mg,0.73mmol)溶于四氢呋喃(5mL)，冷却至-25℃，加入1N LiHMDS(0.73ml,0.73mmol)，于-25℃下反应3h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-1反应完全，减压蒸干溶剂，残留物经柱层析分离(二氯甲烷：甲醇＝150：1)，得白色固体52mg，产率：25.1％，m.p.167.9～169.2℃。Compound VIII-1 (200 mg, 0.48 mmol) and 1-methyl-1H-pyrazol-5-amine (IX-2) (71 mg, 0.73 mmol) were dissolved in tetrahydrofuran (5 mL), cooled to -25°C, and added 1N LiHMDS (0.73ml, 0.73mmol), react at -25 ℃ for about 3h, wait for TLC (dichloromethane: methanol = 25:1) to detect the raw material VIII-1 reaction is complete, evaporate the solvent under reduced pressure, the residue is passed through the column Chromatographic separation (dichloromethane: methanol = 150:1), to obtain 52 mg of white solid, yield: 25.1%, mp 167.9 ~ 169.2 ℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.36(s,2H,CH 2),3.84(s,3H,CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.58-8.46 (m, 2H, ArH), 8.30 (d, J=7.3 Hz, 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, CH 2 ), 4.36 (s, 2H, CH 2 ), 3.84 (s, 3H, CH 3 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75MHz, CDCl 3 ) 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 C 23H 20ClN 6O:431.1382；Found:431.1389. HRMS(ESI): m/z[M+H] + .Calcd for C 23 H 20 ClN 6 O: 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. 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.
实施例5Example 5
2-(3-氯苄基)-6-(2-((1-甲基-1H-吡唑-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-5)的合成2-(3-chlorobenzyl)-6-(2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)isoindolin-1-one (I- 5) Synthesis
将化合物VIII-1(200mg,0.48mmol)和1-甲基-1H-吡唑-4-胺(IX-3)(71mg,0.73mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测VIII-1反应完全，将反应液冷却至室温，减压蒸干溶剂，经柱层析分离(二氯甲烷：甲醇＝150：1)，得淡黄色固体107mg，产率：51.7％，m.p.160.1～161.3℃。Compound VIII-1 (200 mg, 0.48 mmol) and 1-methyl-1H-pyrazole-4-amine (IX-3) (71 mg, 0.73 mmol) were dissolved in sec-butanol (3 mL), and the tube was heated to 125 The reaction was carried out at ℃ for about 12h. After TLC (dichloromethane: methanol = 25:1) detection VIII-1 reaction was complete, the reaction solution was cooled to room temperature, the solvent was evaporated under reduced pressure, and separated by column chromatography (dichloromethane: methanol = 150:1), to obtain 107mg of light yellow solid, yield: 51.7%, mp160.1 ~ 161.3 ℃.
1H-NMR(300MHz,DMSO-d 6)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,CH 2),4.48(s,2H,CH 2),3.82(s,3H,CH 3). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.56 (s, 1H, NH), 8.5-8.47 (m, 2H, ArH), 8.37 (d, J=8.1 Hz, 1H, ArH) , 7.90 (s, 1H, ArH), 7.72 (d, J = 7.6 Hz, 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, CH 2 ), 4.48 (s, 2H, CH 2 ), 3.82 (s, 3H, CH 3 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 23H 20ClN 6O:431.1382；Found:431.1386. HRMS(ESI): m/z[M+H] + .Calcd for C 23 H 20 ClN 6 O: 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. IR(cm -1 ):3446.28,1682.78,1625.78,1573.42,1433.39,1199.84,798.84,769.01,710.30,611.06.
实施例6Example 6
2-(3-氯-4-氟苄基)-6-(2-((1-甲基-1H-吡唑-5-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-6)的合成2-(3-chloro-4-fluorobenzyl)-6-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)isoindoline-1- Synthesis of ketone (I-6)
将化合物VIII-3(207mg,0.48mmol)和1-甲基-1H-吡唑-5-胺(IX-2)(71mg,0.73mmol)溶于四氢呋喃(5mL)，冷却至-25℃，加入1N LiHMDS(0.73mL,0.73mmol)，于-25℃下反应3h左右，TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-3反应完全，减压蒸干溶剂，柱层析分离(二氯甲烷：甲醇＝150：1)，得淡黄色固体62mg，产率：28.8％，m.p.108.1～109.2℃。Compound VIII-3 (207 mg, 0.48 mmol) and 1-methyl-1H-pyrazol-5-amine (IX-2) (71 mg, 0.73 mmol) were dissolved in tetrahydrofuran (5 mL), cooled to -25°C, and added 1N LiHMDS (0.73mL, 0.73mmol), reaction at -25 ℃ for about 3h, TLC (dichloromethane: methanol = 25:1) detection of raw materials VIII-3 complete reaction, evaporation of the solvent under reduced pressure, column chromatography ( Dichloromethane: methanol=150:1), 62 mg of light yellow solid was obtained, yield: 28.8%, mp108.1-109.2°C.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.36(s,2H,CH 2),3.83(s,3H,CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.55-8.45 (m, 2H, ArH), 8.29 (d, J=8.0 Hz, 1H, ArH), 7.54-7.46 (m, 2H, ArH) , 7.38 (d, J = 6.2 Hz, 1H, ArH), 7.30 (d, J = 4.7 Hz, 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, CH 2 ), 4.36 (s, 2H, CH 2 ), 3.83 (s, 3H, CH 3 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) δ (ppm): 167.94, 164.44, 160.22, 159.86 (d, J = 92.7 Hz), 159.03, 143.64, 138.32, 137.27, 137.06, 133.87 (d, J = 3.8 Hz) ,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 C 23H 19ClFN 6O:449.1287；Found:449.1290. HRMS(ESI): m/z[M+H] + .Calcd for C 23 H 19 ClFN 6 O: 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. 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.
实施例7Example 7
2-(1-(3,4-二氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-7)的合成2-(1-(3,4-dichlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl) Synthesis of isoindolin-1-one (I-7)
2-甲基-5-(2-(甲硫基)嘧啶-4-基)苯甲酸甲酯(IV)的合成Synthesis of 2-methyl-5-(2-(methylthio)pyrimidin-4-yl)benzoic acid methyl ester (IV)
将4-氯-2-(甲硫基)嘧啶(XII)(19.8g,123.4mmol)，2-甲基-5-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊烷-2-基)苯甲酸甲酯(III)(34.1g,123.4mmol)，四三苯基膦钯(7.1g,6.17mmol)和碳酸钠(26.2g,246.8mmol)溶于甲苯(90mL)、甲醇(30mL)和水(30mL)组成的混合溶剂中，氮气保护下，70℃反应8h左右，待TLC(石油醚：乙酸乙酯＝10:1)检测原料反应完全，冷却至室温，加入乙酸乙酯(100mL)，静置分层后，减压蒸干溶剂得残留物，柱层析(石油醚：乙酸乙酯＝25：1)分离，得白色固体29.8g，产率：88.0％，m.p.66.2～67.3℃。Combine 4-chloro-2-(methylthio)pyrimidine (XII) (19.8 g, 123.4 mmol), 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2- Dioxaborolan-2-yl)benzoic acid methyl ester (III) (34.1g, 123.4mmol), tetratriphenylphosphine palladium (7.1g, 6.17mmol) and sodium carbonate (26.2g, 246.8mmol) ) Dissolved in a mixed solvent composed of toluene (90mL), methanol (30mL) and water (30mL), under nitrogen protection, react at 70 ℃ for about 8h, wait for TLC (petroleum ether: ethyl acetate = 10:1) to detect the raw material reaction Completely, cool to room temperature, add ethyl acetate (100 mL), and after standing to separate layers, evaporate the solvent under reduced pressure to obtain a residue, which is separated by column chromatography (petroleum ether: ethyl acetate = 25: 1) to obtain a white solid 29.8 g, yield: 88.0%, mp66.2～67.3℃.
1H-NMR(300MHz,CDCl 3)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,OCH 3),2.65(s,3H,SCH 3),2.64(s,3H,CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.58 (d, J=1.9 Hz, 1H, ArH), 8.54 (d, J=5.3 Hz, 1H, ArH), 8.16 (dd, J=8.0 , 2.0Hz, 1H, ArH), 7.40-7.35 (m, 2H, ArH), 3.92 (s, 3H, OCH 3 ), 2.65 (s, 3H, SCH 3 ), 2.64 (s, 3H, CH 3 ).
2-甲基-5-(2-(甲磺酰基)嘧啶-4-基)苯甲酸甲酯(V)的合成Synthesis of methyl 2-methyl-5-(2-(methylsulfonyl)pyrimidin-4-yl)benzoate (V)
将化合物IV(24.2g,88.3mmol)溶于二氯甲烷(150mL)，冰浴下分批加入间氯过氧苯甲酸(38.1g,220.8mmol)，室温搅拌2h左右，待TLC(二氯甲烷：甲醇＝20:1)检测原料IV反应完全，加入二氯甲烷(100mL)稀释，加入饱和硫代硫酸钠溶液(100mL)搅拌20min，静置分层，水层用二氯甲烷萃取(100mL x3)，合并有机层，用饱和碳酸钠溶液(100mL)洗涤，再用饱和氯化钠溶液(100mL)洗涤，无水硫酸钠干燥，过滤，减压蒸干溶剂，得淡黄色固体25.7g，产率：95.0％，m.p.121.8～123.3℃。Compound IV (24.2g, 88.3mmol) was dissolved in dichloromethane (150mL), m-chloroperoxybenzoic acid (38.1g, 220.8mmol) was added in portions under ice bath, stirred at room temperature for about 2h, until TLC (dichloromethane) : Methanol = 20:1) Test the raw material IV reaction is complete, add dichloromethane (100mL) to dilute, add saturated sodium thiosulfate solution (100mL) and stir for 20min, let stand for separation, the aqueous layer is extracted with dichloromethane (100mL x 3) ), the organic layers were combined, washed with saturated sodium carbonate solution (100 mL), and then with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to obtain 25.7 g of light yellow solid. Rate: 95.0%, mp121.8～123.3℃.
1H-NMR(300MHz,CDCl 3)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,OCH 3),3.42(s,3H,SCH 3),2.66(s,3H,CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.91 (d, J=5.3 Hz, 1H, ArH), 8.62 (d, J=1.9 Hz, 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, OCH 3 ), 3.42 (s ,3H,SCH 3 ),2.66(s,3H,CH 3 ).
2-(溴甲基)-5-(2-(甲磺酰基基)嘧啶-4-基)苯甲酸甲酯(VI)的合成Synthesis of methyl 2-(bromomethyl)-5-(2-(methylsulfonyl)pyrimidin-4-yl)benzoate (VI)
将原料V(23.6g,77.0mmol)和NBS(15.1g,84.7mmol)溶于苯(150mL)，加入偶氮二异丁氰(2.5g,15.4mmol)，80℃反应3h左右，TLC(石油醚：乙酸乙酯＝1:1)检测原料反应完全，冷却至室温，浓缩得红色油状物，不经处理直接投下一步。Dissolve raw materials V (23.6g, 77.0mmol) and NBS (15.1g, 84.7mmol) in benzene (150mL), add azobisisobutyl cyanide (2.5g, 15.4mmol), react at 80℃ for about 3h, TLC (Petroleum Ether: ethyl acetate = 1:1) The reaction of the raw materials was detected to be complete, cooled to room temperature, and concentrated to obtain a red oil, which was directly sent to the next step without treatment.
2-(1-(3,4-二氯苯基)-2-羟基乙基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-4)的合成2-(1-(3,4-dichlorophenyl)-2-hydroxyethyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII- 4) Synthesis
将化合物VI(1g,2.6mmol)，2-氨基-2-(3,4-二氯苯基)乙基-1-醇(VII-1)(533mg,2.6mmol)和三乙胺(540μL,3.9mmol)溶于甲醇(10mL)，回流反应6h左右，TLC(二氯甲烷：甲醇＝20:1)检测原料反应完全，冰浴冷却至10℃以下，搅拌30min，有固体析出，抽滤，得白色固体694mg，产率：55.8％，m.p.203.9～204.9℃。Compound VI (1 g, 2.6 mmol), 2-amino-2-(3,4-dichlorophenyl)ethyl-1-ol (VII-1) (533 mg, 2.6 mmol) and triethylamine (540 μL, 3.9mmol) was dissolved in methanol (10mL), the reaction was refluxed for about 6h, TLC (dichloromethane: methanol = 20:1) detected that the reaction of the raw materials was complete, cooled to below 10°C in an ice bath, stirred for 30min, solid precipitated, suction filtered, 694 mg of white solid was obtained, yield: 55.8%, mp 203.9-204.9°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2C H),4.82(d,J＝19.0Hz,1H,CONC H 2 ),4.59(d,J＝18.4Hz,1H,CONC H 2 ),4.24-4.02(m,2H,HOC H 2 ),3.55(s,3H,SO 2C H 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.12 (d, J=5.1 Hz, 1H, ArH), 8.59 (s, 1H, ArH), 8.52 (d, J=8.4 Hz, 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 = 9.6 Hz, 1H, ArH), 5.50-5.37 (m, 1H, HOCH 2 C H ), 4.82 (d, J = 19.0 Hz, 1H, CONC H 2 ), 4.59 (d, J = 18.4 Hz, 1H, CONC H 2 ), 4.24-4.02 (m, 2H, HOC H 2 ), 3.55 (s, 3H, SO 2 C H 3 ).
2-(1-(3,4-二氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-7)的合成2-(1-(3,4-dichlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl) Synthesis of isoindolin-1-one (I-7)
将化合物VIII-4(229mg,0.48mmol)和4-氨基四氢吡喃(IX-1)(74mg,0.73mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测反应完全，将反应液冷却至室温，减压蒸干溶剂，经柱层析分离(二氯甲烷：甲醇＝250：1～50：1)，得白色固体124mg，产率：51.7％，m.p.217.9～219.3℃。Compound VIII-4 (229mg, 0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg, 0.73mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol = 25:1) detects the completion of the reaction, the reaction solution was cooled to room temperature, the solvent was evaporated under reduced pressure, and separated by column chromatography (dichloromethane: methanol = 250: 1 ~ 50: 1), 124 mg of white solid was obtained, yield: 51.7%, mp217.9-219.3°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2C H),4.72(d,J＝17.9Hz,1H,CONC H 2 ),4.48(d,J＝18.2Hz,1H,CONC H 2 ),4.12-3.96(m,3H,HOC H 2 ,NHC H),3.91-3.88(m,2H,O(C H 2) 2 ),3.59-3.40(m,2H,O(C H 2) 2 ),1.61-1.57(m,2H,NHC H 2 ),1.51-1.51(m,2H,NHC H 2 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 8.42 (s, 1H, ArH), 8.39 (d, J=5.2 Hz, 1H, ArH), 8.34 (d, J=8.3 Hz, 1H , ArH), 7.72 (d, J = 8.2 Hz, 1H, ArH), 7.64 (d, J = 1.4 Hz, 1H, ArH), 7.61 (d, J = 8.4 Hz, 1H, ArH), 7.35 (dd, J = 8.3, 1.4 Hz, 1H, ArH), 7.31 (s, 1H, NH), 7.26 (d, J = 5.3 Hz, 1H, ArH), 5.36 (t, J = 6.6 Hz, 1H, HOCH 2 C H ), 4.72 (d, J = 17.9 Hz, 1H, CONC H 2 ), 4.48 (d, J = 18.2 Hz, 1H, CONC H 2 ), 4.12-3.96 (m, 3H, HOC H 2 , NHC H ), 3.91-3.88(m, 2H, O(C H 2 ) 2 ), 3.59-3.40(m, 2H, O(C H 2 ) 2 ), 1.61-1.57(m, 2H, NHC H 2 ), 1.51-1.51 (m,2H,NHC H 2 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 25H 25Cl 2N 4O 3:499.1298；Found:499.1295. HRMS(ESI): m/z[M+H] + .Calcd for C 25 H 25 Cl 2 N 4 O 3 : 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. 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.
实施例8Example 8
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-8)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindole Synthesis of quinolin-1-one (I-8)
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-5)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII-5) synthesis
将化合物VI(9.63g,25.0mmol)，2-氨基-2-(3-氯苯基)乙基-1-醇(VII-2)(4.29g,25.0mmol)和三乙胺(10.4mL,74.9mmol)溶于甲醇(100mL)，回流反应6h左右，待TLC(二氯甲烷：甲醇＝20:1)检测原料反应完全，冰浴冷却至10℃以下，搅拌30min，有固体析出，抽滤，得白色固体7.2g，产率：64.9％，m.p.166.2～167.4℃。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) was dissolved in methanol (100mL), and the reaction was refluxed for about 6h. After TLC (dichloromethane: methanol = 20:1), the reaction of the raw materials was detected. The ice bath was cooled to below 10°C, stirred for 30min, solid precipitated, and filtered , To obtain 7.2g white solid, yield: 64.9%, mp 166.2 ~ 167.4 ℃.
1H-NMR(300MHz,CDCl 3)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,C HCH 2OH),4.63(d,J＝17.8Hz,1H,C H 2 ),4.43-4.32(m,2H,HOC H 2 CH),4.28(dd,J＝11.9,4.4Hz,1H,C H 2 ),3.49(s,3H,SO 2C H 3 ). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 9.01 (d, J=5.3 Hz, 1H, ArH), 8.49 (s, 1H, ArH), 8.44 (dd, J=8.0, 1.5 Hz, 1H , ArH), 8.01 (d, J = 5.3 Hz, 1H, ArH), 7.58 (d, J = 8.0 Hz, 1H, ArH), 7.50-7.31 (m, 3H, ArH), 7.28-7.26 (m, 1H , ArH), 5.40 (dd, J = 7.9, 4.4 Hz, 1H, C H CH 2 OH), 4.63 (d, J = 17.8 Hz, 1 H, C H 2 ), 4.43-4.32 (m, 2H, HOC H 2 CH), 4.28 (dd, J = 11.9, 4.4 Hz, 1H, C H 2 ), 3.49 (s, 3H, SO 2 C H 3 ).
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-8)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindole Synthesis of quinolin-1-one (I-8)
将化合物VIII-5(213mg,0.48mmol)和4-氨基四氢吡喃(IX-1)(74mg,0.73mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-5反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析分离(二氯甲烷：甲醇＝250：1～50：1)，得淡黄色固体117mg，产率：52.4％，m.p.141.0～142.3℃。Compound VIII-5 (213mg, 0.48mmol) and 4-aminotetrahydropyran (IX-1) (74mg, 0.73mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol = 25:1) detects that the raw material VIII-5 is completely reacted. The reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, and separated by column chromatography (dichloromethane: methanol = 250: 1-50: 1), 117mg of light yellow solid was obtained, yield: 52.4%, mp141.0～142.3℃.
1H-NMR(300MHz,CDCl 3)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,HOCH 2C H),4.51(d,J＝17.0Hz,1H,CONC H 2 ),4.37-4.19(m,4H,CONC H 2 ,HOC H 2 ,NHC H),4.04-4.01(m,2H,O(C H 2) 2 ),3.63-3.57(m,2H,O(C H 2) 2 ),2.11-2.08(m,2H,NHC H 2 ),1.68-1.64(m,2H,NHC H 2 ). 1 H-NMR (300 MHz, CDCl 3 ) 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, HOCH 2 C H ), 4.51 (d, J=17.0 Hz,1H,CONC H 2 ),4.37-4.19(m,4H,CONC H 2 ,HOC H 2 ,NHC H ),4.04-4.01(m,2H,O(C H 2 ) 2 ),3.63-3.57( m,2H,O(C H 2 ) 2 ),2.11-2.08(m,2H,NHC H 2 ),1.68-1.64(m,2H,NHC H 2 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 25H 26ClN 4O 3:465.1688；Found:465.1688. HRMS(ESI): m/z[M+H] + .Calcd for C 25 H 26 ClN 4 O 3 : 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. 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.
实施例9Example 9
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((1-甲基-1H-吡唑-5-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-9)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)iso Synthesis of indolin-1-one (I-9)
2-(2-((叔丁基二甲基硅基)氧基)-1-(3-氯苯基)乙基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(X-1)的合成2-(2-((tert-butyldimethylsilyl)oxy)-1-(3-chlorophenyl)ethyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)iso Synthesis of indolin-1-one (X-1)
将化合物VIII-5(1.0g,2.26mmol)和咪唑(0.31g,4.52mmol)溶于二氯甲烷(15mL)，加入TBSCl(0.68g,4.52mmol)，于35℃搅拌反应8h左右，TLC(二氯甲烷:甲醇＝20：1)，检测原料VIII-5反应完全，用饱和碳酸氢钠(6mL)洗涤，再用饱和氯化钠溶液(6mL)洗涤，无水硫酸钠干燥，过滤，减压蒸干溶剂，加入石油醚(6mL)打浆，过滤，得白色固体1.20g，收率95.1％，m.p.131.7～133.2℃。Compound VIII-5 (1.0g, 2.26mmol) and imidazole (0.31g, 4.52mmol) were dissolved in dichloromethane (15mL), TBSCl (0.68g, 4.52mmol) was added, the reaction was stirred at 35 ℃ for about 8h, TLC ( Dichloromethane: methanol = 20:1), detecting that the reaction of the raw material VIII-5 is complete, washed with saturated sodium bicarbonate (6 mL), then with saturated sodium chloride solution (6 mL), dried over anhydrous sodium sulfate, filtered, and reduced The solvent was evaporated under pressure, added with petroleum ether (6mL) to beat, and filtered to obtain a white solid 1.20g, yield 95.1%, mp 131.7～133.2°C.
1H-NMR(300MHz,CDCl 3)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,C HCH 2OSi),4.70(d,J＝17.9Hz,1H,C H 2 ),4.36(d,J＝18.0Hz,1H,C H 2 ),4.28-4.21(m,2H,SiOC H 2 CH),3.46(s,3H,SO 2C H 3 ),0.84(s,9H,SiC(C H 3) 3 ),0.07(s,3H,SiC H 3 ),0.05(s,3H,SiC H 3 ). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.99 (d, J=5.3 Hz, 1H, ArH), 8.58-8.49 (m, 2H, ArH), 8.02 (d, J=5.4 Hz, 1H , ArH), 7.63 (d, J = 7.9 Hz, 1H, ArH), 7.44 (s, 1H, ArH), 7.29 (d, J = 6.5 Hz, 3H, ArH), 5.58 (t, J = 5.4 Hz, 1H, CH H 2 OSi), 4.70 (d, J = 17.9 Hz, 1H, CH 2 ), 4.36 (d, J = 18.0 Hz, 1H, CH 2 ), 4.28-4.21 (m, 2H, SiOC H 2 CH), 3.46 (s, 3H, SO 2 C H 3 ), 0.84 (s, 9H, SiC (C H 3 ) 3 ), 0.07 (s, 3H, SiC H 3 ), 0.05 (s, 3H, SiC H 3 ).
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((1-甲基-1H-吡唑-5-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-9)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)iso Synthesis of indolin-1-one (I-9)
将化合物X-1(200mg,0.36mmol)和1-甲基-1H-吡唑-5-胺(IX-2)(52mg,0.54mmol)溶于无水四氢呋喃(5mL)，冷却至-25℃，加入1N LiHMDS(0.54ml,0.54mmol)，-25℃反应3h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料X-1反应完全，减压蒸干溶剂，加入乙酸乙酯(5mL)溶解，用HCl的乙酸乙酯溶液调节pH＝2～3，继续搅拌反应3h左右，TLC(二氯甲烷：甲醇＝25：1)检测中间体反应完全，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体35mg，产率：21.1％，m.p.118.2～119.4℃。Compound X-1 (200 mg, 0.36 mmol) and 1-methyl-1H-pyrazol-5-amine (IX-2) (52 mg, 0.54 mmol) were dissolved in anhydrous tetrahydrofuran (5 mL) and cooled to -25°C , Add 1N LiHMDS (0.54ml, 0.54mmol), react at -25℃ for about 3h, wait for TLC (dichloromethane: methanol = 25:1) to detect the completion of the raw material X-1, evaporate the solvent under reduced pressure, add ethyl acetate (5mL) Dissolve, adjust the pH=2～3 with HCl ethyl acetate solution, continue to stir the reaction for about 3h, TLC (dichloromethane: methanol=25:1) to detect the completion of the intermediate reaction, evaporate the solvent under reduced pressure, column Chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 50: 1) separation, to obtain a white solid 35mg, yield: 21.1%, mp 118.2 ~ 119.4 ℃.
1H-NMR(300MHz,CDCl 3)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,HOCH 2C H),4.52(d,J＝17.4Hz,1H,CONC H 2 ),4.47-4.12(m,3H,CONC H 2 ,HOC H 2 ),3.81(s,3H,C H 3 ). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.45 (d, J=5.0 Hz, 1H, ArH), 8.32 (s, 1H, ArH), 8.07 (d, J=7.5 Hz, 1H, ArH ), 7.84 (s, 1H, ArH), 7.41-7.35 (m, 3H, ArH), 7.28 (m, 2H, ArH), 7.17 (d, J = 5.0 Hz, 1H, ArH), 6.15 (s, 1H , ArH), 5.41 (s, 1H, HOCH 2 C H ), 4.52 (d, J = 17.4 Hz, 1H, CONC H 2 ), 4.47-4.12 (m, 3H, CONC H 2 , HOC H 2 ), 3.81 (s,3H,C H 3 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 24H 22ClN 6O 2:461.1487；Found:461.1489. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 22 ClN 6 O 2 : 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. 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.
实施例10Example 10
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((1-甲基-1H-吡唑-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-10)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)iso Synthesis of indolin-1-one (I-10)
将化合物VIII-5(160mg,0.36mmol)和1-甲基-1H-吡唑-4-胺(IX-3)(52mg,0.54mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得土黄色固体93mg，产率：56.0％，m.p.218.2～219.4℃。Compound VIII-5 (160 mg, 0.36 mmol) and 1-methyl-1H-pyrazol-4-amine (IX-3) (52 mg, 0.54 mmol) were dissolved in sec-butanol (3 mL), and the tube was heated to 125 The reaction was carried out at ℃ for about 12h. After TLC (dichloromethane: methanol = 25:1), the reaction was detected to be complete. The reaction solution was cooled to room temperature, and the solvent was evaporated under reduced pressure. : 1-50:1) Isolation gave 93 mg of yellowish-yellow solid, yield: 56.0%, mp 218.2-219.4°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2C H),4.73(d,J＝16.7Hz,1H,CONC H 2 ),4.47(d,J＝17.8Hz,1H,CONC H 2 ),4.01(s,2H,HOC H 2 ),3.82(s,3H,CH 3). 1 H-NMR (300 MHz, DMSO-d 6 ) 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, HOCH 2 C H ), 4.73 (d, J = 16.7 Hz, 1H, CONC H 2 ), 4.47 (d, J = 17.8 Hz, 1H, CONC H 2 ), 4.01 (s, 2H, HOC H 2 ), 3.82 (s, 3H, CH 3 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 24H 22ClN 6O 2:461.1487；Found:461.1494. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 22 ClN 6 O 2 : 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. 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.
实施例11Example 11
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(异丙基氨基)嘧啶-4-基)异吲哚啉-1-酮(I-11)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(isopropylamino)pyrimidin-4-yl)isoindolin-1-one (I-11) Synthesis
将化合物VIII-5(200mg,0.45mmol)和异丙胺(IX-3)(40mg,0.68mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-5反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体96mg，产率：50.4％，m.p.140.1～142.4℃。Compound VIII-5 (200mg, 0.45mmol) and isopropylamine (IX-3) (40mg, 0.68mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, after TLC (dichloromethane : Methanol = 25:1) Test the raw material VIII-5 reaction is complete, the reaction solution is cooled to room temperature, the solvent is evaporated to dryness under reduced pressure, column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 50: 1) Separation gave 96 mg of white solid, yield: 50.4%, mp 140.1-142.4°C.
1H-NMR(300MHz,CDCl 3)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,HOCH 2C H,N HCH),4.52(d,J＝17.5Hz,1H,CONC H 2 ),4.31-4.25(m,4H,CONC H 2 ,HOC H 2 ,NHC H),1.30(d,J＝6.5Hz,6H,NHCH( CH 3) 2 ). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.42 (s, 1H, ArH), 8.36-8.32 (m, 1H, ArH), 8.24 (d, J=7.6 Hz, 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, HOCH 2 C H , N H CH), 4.52 (d, J = 17.5 Hz, 1 H, CONC H 2 ), 4.31-4.25 (m, 4H, CONC H 2 , HOC H 2 , NHC H ), 1.30 (d, J = 6.5Hz, 6H, NHCH ( CH 3 ) 2 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 23H 24ClN 4O 2:423.1588；Found:423.1588. HRMS(ESI): m/z[M+H] + .Calcd for C 23 H 24 ClN 4 O 2 : 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. IR (cm -1 ): 3478.65, 3243.37, 1660.99, 1597.27, 1573.23, 1537.51, 1459.55, 1408.28, 1203.39, 1178.35, 1117.90, 773.23.
实施例12Example 12
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(2-羟乙基氨基)嘧啶-4-基)异吲哚啉-1-酮(I-12)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(2-hydroxyethylamino)pyrimidin-4-yl)isoindolin-1-one (I- 12) Synthesis
将化合物VIII-5(300mg,0.68mmol)和乙醇胺(IX-5)(124mg,2.03mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-5反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体160mg，产率：55.7％，m.p.157.5～160.1℃。Compound VIII-5 (300mg, 0.68mmol) and ethanolamine (IX-5) (124mg, 2.03mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, after TLC (dichloromethane: Methanol = 25:1) The reaction of the raw material VIII-5 was detected to be complete. The reaction solution was cooled to room temperature, the solvent was evaporated under reduced pressure, and separated by column chromatography (eluent: dichloromethane: methanol = 200: 1-50: 1). , To obtain 160mg of white solid, yield: 55.7%, mp 157.5 ~ 160.1 ℃.
1H-NMR(300MHz,CD 3OD)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,HOCH 2C H),4.80-4.72(m,1H,CONC H 2 ),4.44(d,J＝18.1Hz,1H,CONC H 2 ),4.27-4.17(m,2H,HO CH 2 CH),3.77-3.76(m,2H,HO CH 2 CH 2NH),3.67-3.62(m,2H,HOCH 2 CH 2 NH). 1 H-NMR (300 MHz, CD 3 OD) 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,HOCH 2 C H ),4.80-4.72(m,1H,CONC H 2 ),4.44(d, J = 18.1 Hz, 1H, CONC H 2 ), 4.27-4.17 (m, 2H, HO CH 2 CH), 3.77-3.76 (m, 2H, HO CH 2 CH 2 NH), 3.67-3.62 (m, 2H, HOCH 2 CH 2 NH).
13C-NMR(75MHz,CD 3OD)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. 13 C-NMR (75MHz, CD 3 OD) 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 C 22H 22ClN 4O 3:425.1380；Found:425.1376. HRMS(ESI): m/z[M+H] + .Calcd for C 22 H 22 ClN 4 O 3 : 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. 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.
实施例13Example 13
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(3-羟丙基氨基)嘧啶-4-基)异吲哚啉-1-酮(I-13)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(3-hydroxypropylamino)pyrimidin-4-yl)isoindolin-1-one (I- 13) Synthesis
将化合物VIII-5(300mg,0.68mmol)和丙醇胺(IX-6)(152mg,2.03mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-5反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体130mg，产率：43.6％，m.p.64.2～66.3℃。Compound VIII-5 (300mg, 0.68mmol) and propanolamine (IX-6) (152mg, 2.03mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, TLC (dichloro Methane: methanol = 25:1) Test the raw material VIII-5 reaction is complete, cool the reaction solution to room temperature, evaporate the solvent under reduced pressure, column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 50: 1 ) Separation to obtain 130 mg of white solid, yield: 43.6%, mp 64.2 ~ 66.3 ℃.
1H-NMR(300MHz,CDCl 3)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, HNCH 2),5.44-5.40(m,1H,HOCH 2 CH),4.59(d,J＝17.5Hz,1H,CONC H 2 ),4.38-4.21(m,3H,CONC H 2 ,HO CH 2 CH),4.02(brs,2H, HOCH 2CH, HOCH 2CH 2),3.75-3.68(m,4H,HO CH 2 CH 2 CH 2 NH),1.88-1.85(m,2H,HOCH 2 CH 2 CH 2NH). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.37 (s, 1H, ArH), 8.28 (s, 1H, ArH), 8.17 (d, J=7.7 Hz, 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, HN CH 2 ), 5.44-5.40(m, 1H, HOCH 2 CH ), 4.59 (d, J = 17.5 Hz, 1H, CONC H 2 ), 4.38-4.21 (m, 3H, CONC H 2 , HO CH 2 CH), 4.02 (brs, 2H, HO CH 2 CH, HO CH 2 CH 2 ), 3.75-3.68 (m, 4H, HO CH 2 CH 2 CH 2 NH), 1.88-1.85 (m, 2H, HOCH 2 CH 2 CH 2 NH).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 23H 24ClN 4O 3:439.1537；Found:439.1532. HRMS(ESI): m/z[M+H] + .Calcd for C 23 H 24 ClN 4 O 3 : 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. IR (cm -1 ): 3349.85, 1668.07, 1572.74, 1493.88, 1453.98, 1407.24, 1342.27, 1266.70, 1201.84, 1067.90, 806.12, 770.92, 695.77, 613.93.
实施例14Example 14
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(环己基氨基)嘧啶-4-基)异吲哚啉-1-酮(I-14)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(cyclohexylamino)pyrimidin-4-yl)isoindolin-1-one (I-14) synthesis
将化合物VIII-5(400mg,0.90mmol)和环己胺(IX-7)(268mg,2.70mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-5反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体180mg，产率：43.2％，m.p.150.4～151.8℃。Compound VIII-5 (400mg, 0.90mmol) and cyclohexylamine (IX-7) (268mg, 2.70mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, TLC (dichloro Methane: methanol = 25:1) Test the raw material VIII-5 reaction is complete, cool the reaction solution to room temperature, evaporate the solvent under reduced pressure, column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 50: 1 ) Separation to obtain 180 mg of white solid, yield: 43.2%, mp 150.4 ~ 151.8 ℃.
1H-NMR(300MHz,CDCl 3)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,HOCH 2 CH),4.73(d,J＝17.7Hz,1H,CONC H 2 ),4.45-4.23(m,4H,CONC H 2 ,HO CH 2 CH,CH 2 CHNH),4.02(s,1H, HOCH 2CH),2.11-2.04(m,2H,cyclohexyl-H),1.91-1.83(m,2H,cyclohexyl-H),1.68-1.36(m,6H,cyclohexyl-H). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.37 (s, 1H, ArH), 8.25 (d, J=7.8 Hz, 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,HOCH 2 CH ),4.73(d,J=17.7Hz,1H,CONC H 2 ),4.45-4.23(m,4H,CONC H 2 ,HO CH 2 CH,CH 2 CH NH),4.02(s ,1H, HO CH 2 CH),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,CDCl 3)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 13 C-NMR (75 MHz, CDCl 3 ) 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 C 26H 28ClN 4O 2:463.1901；Found:463.1896. HRMS(ESI): m/z[M+H] + .Calcd for C 26 H 28 ClN 4 O 2 : 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. 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.
实施例15Example 15
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((4-羟基环己基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-15)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((4-hydroxycyclohexyl)amino)pyrimidin-4-yl)isoindolin-1-one ( I-15) Synthesis
将化合物VIII-5(400mg,0.90mmol)和4-氨基环己醇(IX-8)(311mg,2.70mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-5反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体210mg，产率：48.7％，m.p.170.3～171.6℃。Compound VIII-5 (400mg, 0.90mmol) and 4-aminocyclohexanol (IX-8) (311mg, 2.70mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, until TLC (Dichloromethane: methanol = 25:1) Test the raw material VIII-5 reaction is complete, the reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 50:1) Separation to obtain 210 mg of white solid, yield: 48.7%, mp170.3～171.6℃.
1H-NMR(300MHz,CDCl 3)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,HOCH 2 CH),4.57(d,J＝17.5Hz,1H,CONC H 2 ),4.38-4.22(m,3H,CON CH 2 ,HO CH 2 CH),3.99-3.96(m,1H,(CH 2) 2 CHOH),3.78-3.68(m,1H,(CH 2) 2 CHNH),2.23-2.05(m,4H,hydroxycyclohexyl-H),1.59-1.33(m,4H,hydroxycyclohexyl-H). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.42 (s, 1H, ArH), 8.32-8.31 (m, 1H, ArH), 8.24 (d, J=8.0 Hz, 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, HN CH), 5.40-5.36 (m, 1H, HOCH 2 CH ), 4.57 (d, J = 17.5 Hz, 1H, CONC H 2 ), 4.38-4.22 (m, 3H, CON CH 2 , HO CH 2 CH), 3.99-3.96 (m, 1H, (CH 2 ) 2 CH OH), 3.78-3.68 (m, 1H, (CH 2 ) 2 CH NH), 2.23-2.05 (m, 4H, hydroxycyclohexyl-H) ,1.59-1.33(m,4H,hydroxycyclohexyl-H).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 26H 28ClN 4O 3:479.1850；Found:479.1846. HRMS(ESI): m/z[M+H] + .Calcd for C 26 H 28 ClN 4 O 3 : 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. IR (cm -1 ): 3429.06, 2929.86, 2848.98, 1654.59, 1594.29, 1557.12, 1524.49, 1495.10, 1457.75, 1414.30, 1347.78, 1203.31, 1068.26, 808.64, 771.78, 689.80, 612.24.
实施例16Example 16
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((四氢呋喃-3-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-16)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydrofuran-3-yl)amino)pyrimidin-4-yl)isoindolin-1-one ( I-16) Synthesis
将化合物VIII-5(400mg,0.90mmol)和3-氨基四氢呋喃(IX-8)(48mg,1.35mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-5反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体220mg，产率：54.2％，m.p.192.3～193.9℃。Compound VIII-5 (400mg, 0.90mmol) and 3-aminotetrahydrofuran (IX-8) (48mg, 1.35mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, TLC (two Chloromethane: methanol=25:1) Test the raw material VIII-5 reaction is complete, cool the reaction solution to room temperature, evaporate the solvent under reduced pressure, column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 50: 1) Separation to obtain 220 mg of white solid, yield: 54.2%, mp 192.3 ~ 193.9 °C.
1H-NMR(300MHz,CDCl 3)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,HOCH 2C H),4.70-4.68(m,1H,NH CH),4.54(d,J＝17.6Hz,1H,CON CH 2 ),4.34-4.20(m,3H,CON CH 2 ,HO CH 2 ),4.05-4.01(m,2H,O CH 2 ),3.91-3.89(m,1H,O CH 2 ),3.80-3.78(m,1H,O CH 2 ),2.40-2.34(m,1H,CH CH 2 CH 2O),1.98-1.94(m,1H,CH CH 2 CH 2O). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.38-8.34 (m, 2H, ArH), 8.20 (d, J=7.9 Hz, 1H, ArH), 7.43 (d, J=7.7 Hz, 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, NH CH), 5.35-5.30 (m,1H,HOCH 2 C H ),4.70-4.68(m,1H,NH CH ),4.54(d,J=17.6Hz,1H,CON CH 2 ),4.34-4.20(m,3H,CON CH 2 , HO CH 2 ), 4.05-4.01 (m, 2H, O CH 2 ), 3.91-3.89 (m, 1H, O CH 2 ), 3.80-3.78 (m, 1H, O CH 2 ), 2.40-2.34 (m ,1H,CH CH 2 CH 2 O),1.98-1.94(m,1H,CH CH 2 CH 2 O).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 24H 24ClN 4O 3:451.1537；Found:451.1533. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 24 ClN 4 O 3 : 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. 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.
实施例17Example 17
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(2-氨基噻唑)嘧啶-4-基)异吲哚啉-1-酮(I-17)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(2-aminothiazole)pyrimidin-4-yl)isoindolin-1-one (I-17) Synthesis
将2-氨基噻唑(IX-10)(36mg,0.36mmol)溶于无水四氢呋喃(5mL)，冷却至-78℃，加入1N LiHMDS(0.36ml,0.36mmol)，搅拌30min后，将化合物X-1(100mg,0.18mmol)溶于无水四氢呋喃(3mL)，于-78℃下缓慢滴加入上述反应液，-78℃反应1h左右，待TLC(二氯甲烷：甲醇＝20：1)检测原料X-1反应完全，减压蒸干溶剂，加入乙酸乙酯(5mL)溶解，用HCl的乙酸乙酯溶液调节pH＝2～3，继续搅拌反应3h左右，待TLC(二氯甲烷：甲醇＝15：1)检测中间体反应完，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～30：1)分离，得金黄色固体目标化合物盐酸盐45mg，产率：50.0％，m.p.>220.1℃。Dissolve 2-aminothiazole (IX-10) (36 mg, 0.36 mmol) in anhydrous tetrahydrofuran (5 mL), cool to -78°C, add 1N LiHMDS (0.36 ml, 0.36 mmol), and after stirring for 30 min, transfer compound X- 1 (100mg, 0.18mmol) dissolved in anhydrous tetrahydrofuran (3mL), slowly add the above reaction solution dropwise at -78 ℃, react at -78 ℃ for about 1h, wait for TLC (dichloromethane: methanol = 20: 1) detection of raw materials After the X-1 reaction is complete, evaporate the solvent under reduced pressure, add ethyl acetate (5 mL) to dissolve it, adjust the pH = 2 to 3 with HCl in ethyl acetate, and continue to stir the reaction for about 3 h, until TLC (dichloromethane: methanol = 15:1) After the detection of the intermediate reaction is completed, the solvent is evaporated to dryness under reduced pressure, and the column chromatography (eluent: dichloromethane: methanol = 200: 1 to 30: 1) is separated to obtain the target compound hydrochloride as a golden yellow solid 45mg , Yield: 50.0%, mp>220.1℃.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),4.76(d,J＝17.9Hz,1H,CON CH 2 ),4.49(d,J＝18.2Hz,1H,CON CH 2 ),4.07-4.01(m,2H,HO CH 2 CH). 1 H-NMR (300 MHz, DMSO-d 6 ) 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, HOCH 2 CH ), 4.76 (d, J = 17.9 Hz, 1H, CON CH 2 ), 4.49 (d, J = 18.2 Hz, 1H, CON CH 2 ), 4.07-4.01 (m, 2H, HO CH 2 CH).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 23H 19ClN 5O 2S:464.0948；Found:464.0947. HRMS(ESI): m/z[M+H] + .Calcd for C 23 H 19 ClN 5 O 2 S: 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. 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.
实施例18Example 18
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((1-甲基-1H-吡唑-3-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-18)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((1-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)iso Synthesis of Indoline-1-one (I-18)
将N-甲基-3-氨基吡唑(IX-11)(35mg,0.36mmol)溶于无水四氢呋喃(5mL)，冷却至-78℃，加入1N LiHMDS(0.36mL,0.36mmol)，搅拌30min，将化合物X-1(100mg,0.18mmol)溶于无水四氢呋喃(3mL)，于-78℃下缓慢滴加入上述反应液，-78℃反应1h左右，待TLC(二氯甲烷：甲醇＝20：1)检测原料X-1反应完全，减压蒸干溶剂，加入乙酸乙酯(5mL)溶解，用HCl的乙酸乙酯溶液调节pH＝2～3，继续搅拌3h左右，待TLC(二氯甲烷：甲醇＝15：1)检测中间体反应完，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～30：1)分离，得白色固体目标化合物盐酸盐50mg，产率：55.8％，m.p.218.2～219.8℃。Dissolve N-methyl-3-aminopyrazole (IX-11) (35 mg, 0.36 mmol) in anhydrous tetrahydrofuran (5 mL), cool to -78°C, add 1N LiHMDS (0.36 mL, 0.36 mmol), and stir for 30 min , Dissolve compound X-1 (100mg, 0.18mmol) in anhydrous tetrahydrofuran (3mL), slowly add the above reaction solution dropwise at -78℃, react at -78℃ for about 1h, wait for TLC (dichloromethane: methanol = 20 ：1) Detect the completion of the reaction of the raw material X-1, evaporate the solvent under reduced pressure, add ethyl acetate (5mL) to dissolve, adjust the pH=2～3 with ethyl acetate solution of HCl, continue stirring for about 3h, wait for TLC (dichloromethane) Methane: methanol = 15:1) After the detection of the intermediate reaction is completed, the solvent is evaporated under reduced pressure, and the column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 30: 1) is separated to obtain the target compound salt as a white solid 50 mg of acid salt, yield: 55.8%, mp218.2-219.8°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.24(s,1H, HNC),4.72(d,J＝18.7Hz,1H,CON CH 2 ),4.44(d,J＝18.6Hz,1H,CON CH 2 ),4.04-3.97(m,2H,HO CH 2 CH),3.74(s,3H, CH 3 N). 1 H-NMR (300 MHz, DMSO-d 6 ) 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, HOCH 2 CH ) , 5.24 (s, 1H, HN C), 4.72 (d, J = 18.7 Hz, 1H, CON CH 2 ), 4.44 (d, J = 18.6 Hz, 1H, CON CH 2 ), 4.04-3.97 (m, 2H , HO CH 2 CH), 3.74 (s, 3H, CH 3 N).
13C-NMR(75MHz,DMSO-d 6)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 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 24H 22ClN 6O 2:461.1493；Found:461.1483. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 22 ClN 6 O 2 :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. IR(cm -1 ):3416.33,1660.65,1569.89,1404.85,1204.08,1073.47,795.92,771.43,608.16.
实施例19Example 19
2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(1-甲基-1H-四氮唑-5-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-19)的合成2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(1-methyl-1H-tetrazol-5-yl)amino)pyrimidin-4-yl)iso Synthesis of indolin-1-one (I-19)
将化合物X-1(500mg,0.90mmol)和1-甲基-5-氨基四氮唑(IX-12)(228mg,2.30mmol)溶于无水N,N-二甲基甲酰胺(4mL)，冷却至-70℃，加入1N LiHMDS(1.35mL,1.35mmol)，-70℃反应2h左右，待TLC(二氯甲烷：甲醇＝20：1)检测反应完全，减压蒸干溶剂，加入乙酸乙酯(5mL)溶解，用HCl的乙酸乙酯溶液调节pH＝2～3，继续搅拌3h左右，待TLC(二氯甲烷：甲醇＝15：1)检测中间体反应完全，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～30：1)分离，得淡黄色固体，目标产物盐酸盐180mg，产率：40.1％，m.p.86.3～88.0℃。Compound X-1 (500 mg, 0.90 mmol) and 1-methyl-5-aminotetrazolium (IX-12) (228 mg, 2.30 mmol) were dissolved in anhydrous N,N-dimethylformamide (4 mL) , Cool to -70℃, add 1N LiHMDS (1.35mL, 1.35mmol), react at -70℃ for about 2h, wait for TLC (dichloromethane: methanol = 20:1) to detect the completion of the reaction, evaporate the solvent under reduced pressure, add acetic acid Dissolve the ethyl ester (5mL), adjust the pH=2～3 with HCl in ethyl acetate, and continue stirring for about 3h. After TLC (dichloromethane: methanol=15:1) detects the completion of the intermediate reaction, the solvent is evaporated under reduced pressure. , Column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 30: 1) separation, to give a pale yellow solid, the target product hydrochloride 180mg, yield: 40.1%, mp 86.3 ~ 88.0 ℃.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.19(s,1H, HNCH),4.73(d,J＝18.2Hz,1H,CON CH 2 ),4.45(d,J＝18.2Hz,1H,CON CH 2 ),4.08-4.02(m,2H,HO CH 2 CH),3.91(s,3H,N CH 3 ) 1 H-NMR (300 MHz, DMSO-d 6 ) 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, HOCH 2 CH ), 5.19 (s, 1H, HN CH), 4.73 (d, J=18.2 Hz, 1H, CON CH 2 ), 4.45 (d, J = 18.2 Hz, 1H, CON CH 2 ), 4.08-4.02 (m, 2H, HO CH 2 CH), 3.91 (s, 3H, N CH 3 )
13C-NMR(75MHz,DMSO-d 6)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 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 22H 20ClN 8O 2:463.1398；Found:463.1397. HRMS(ESI): m/z[M+H] + .Calcd for C 22 H 20 ClN 8 O 2 : 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. 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.
实施例20Example 20
(R)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-20)的合成(R)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl ) Synthesis of Isoindoline-1-one (I-20)
(R)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-6)的合成(R)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII -6) Synthesis
将化合物VI(1.57g,4.08mmol)，(R)-2-氨基-2-(3-氯苯基)乙基-1-醇(VII-3)(700mg,4.08mmol)和三乙胺(1.70mL,12.24mmol)溶于乙腈(30mL)，加热回流反应6h左右，待TLC(二氯甲烷：甲醇＝20:1)检测反应完毕，减压蒸干溶剂，加入甲醇(10mL)冰浴冷却至10℃以下，搅拌30min，有固体析出，抽滤，得白色固体1.03g，产率：56.9％，m.p.78.2～79.8℃。Compound VI (1.57 g, 4.08 mmol), (R)-2-amino-2-(3-chlorophenyl)ethyl-1-ol (VII-3) (700 mg, 4.08 mmol) and triethylamine ( 1.70mL, 12.24mmol) was dissolved in acetonitrile (30mL), heated to reflux for about 6h, after TLC (dichloromethane: methanol = 20:1) detection reaction was completed, the solvent was evaporated under reduced pressure, methanol (10mL) was added to cool in ice bath When the temperature is below 10℃, stir for 30min, solid precipitates out, and filter by suction to obtain 1.03g of white solid, yield: 56.9%, mp 78.2-79.8℃.
1H-NMR(300MHz,DMSO-d 6)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, CHCH 2OH),5.22-5.19(m,1H,CHCH 2 OH),4.78(d,J＝18.6Hz,1H, CH 2 N),4.51(d,J＝18.6Hz,1H, CH 2N)4.08 4.01(m,2H,HO CH 2 CH),3.51(s,3H, CH 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.15 (d, J=5.3 Hz, 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, CH CH 2 OH), 5.22-5.19 (m, 1H, CHCH 2 OH ), 4.78 (d, J = 18.6 Hz, 1H, CH 2 N), 4.51 (d, J = 18.6 Hz, 1H, CH 2 N) 4.08 4.01 (m, 2H, HO CH 2 CH), 3.51 (s, 3H, CH 3 ).
(R)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-20)的合成(R)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl ) Synthesis of Isoindoline-1-one (I-20)
将化合物VIII-6(400mg,0.90mmol)和4-氨基四氢吡喃(IX-1)(182mg,1.80mmol)溶于仲丁醇(5mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-6反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200:1～50:1)分离，得白色固体208mg，产率：49.7％，m.p.83.8～85.7℃。Compound VIII-6 (400mg, 0.90mmol) and 4-aminotetrahydropyran (IX-1) (182mg, 1.80mmol) were dissolved in sec-butanol (5mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol=25:1) detects that the raw material VIII-6 is completely reacted. The reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, and column chromatography (eluent: dichloromethane: methanol=200:1) ~50:1) Separation gave 208 mg of white solid, yield: 49.7%, mp 83.8-85.7°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.27-5.24(m,1H, HOCH 2CH),4.78(d,J＝18.3Hz,1H,CONC H 2 ),4.50(d,J＝18.3Hz,1H,CONC H 2 ),4.09-4.07(m,3H,NH CH,HO CH 2 ),3.96-3.93(m,2H,O(C H 2) 2 ),3.56-3.47(m,2H,O(C H 2) 2 ),1.96-1.92(m,2H,NHCH(C H 2) 2 ),1.66-1.59(m,2H,NHCH(C H 2) 2 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 8.47-8.37 (m, 3H, ArH), 7.77 (d, J = 7.8 Hz, 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,HOCH 2 CH ),5.27-5.24(m,1H, HO CH 2 CH), 4.78 (d, J = 18.3 Hz, 1H, CONC H 2 ), 4.50 (d, J = 18.3 Hz, 1H, CONC H 2 ), 4.09-4.07 (m, 3H, NH CH , HO CH 2 ), 3.96 3.93(m,2H,O(C H 2 ) 2 ),3.56-3.47(m,2H,O(C H 2 ) 2 ),1.96-1.92(m,2H,NHCH(C H 2 ) 2 ),1.66 -1.59(m, 2H, NHCH(C H 2 ) 2 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 25H 26ClN 4O 3:465.1693；Found:465.1687. HRMS(ESI): m/z[M+H] + .Calcd for C 25 H 26 ClN 4 O 3 : 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. 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.
实施例21Example 21
(R)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((1-甲基-1H-吡唑-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-21)的合成(R)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidine-4 -Yl) isoindolin-1-one (I-21) synthesis
将化合物VIII-6(400mg,0.90mmol)和1-甲基-1H-吡唑-4-胺(IX-3)(175mg,1.80mmol)溶于仲丁醇(5mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝20：1)检测原料VIII-6反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得金黄色固体100mg，产率：24.1％，m.p.229.7～231.6℃。Compound VIII-6 (400 mg, 0.90 mmol) and 1-methyl-1H-pyrazole-4-amine (IX-3) (175 mg, 1.80 mmol) were dissolved in sec-butanol (5 mL), and the tube was heated to 125 The reaction was carried out at ℃ for about 12h. After TLC (dichloromethane: methanol = 20:1), the raw material VIII-6 was detected to be complete. The reaction solution was cooled to room temperature, the solvent was evaporated under reduced pressure, and column chromatography (eluent: dichloromethane : Methanol = 200: 1 to 50: 1) Separation to obtain 100 mg of golden yellow solid, yield: 24.1%, mp 229.7 to 231.6 °C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.27-5.24(m,1H, HOCH 2CH),4.79(d,J＝18.0Hz,1H,CON CH 2 ),4.52(d,J＝18.2Hz,1H,CON CH 2 ),4.14-4.07(m,2H,HO CH 2 ),3.88(s,3H,N CH 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) 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,HOCH 2 CH ),5.27-5.24(m,1H, HO CH 2 CH),4.79(d,J=18.0Hz,1H,CON CH 2 ),4.52(d,J=18.2Hz,1H , CON CH 2 ), 4.14-4.07 (m, 2H, HO CH 2 ), 3.88 (s, 3H, N CH 3 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 24H 22ClN 6O 2:461.1493；Found:461.1492. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 22 ClN 6 O 2 : 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. 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.
实施例22Example 22
(S)-2-(2-羟基-1-苯乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-22)的合成(S)-2-(2-Hydroxy-1-phenethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindoline- Synthesis of 1-ketone (I-22)
(S)-2-(2-羟基-1-苯乙基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-7)的合成Synthesis of (S)-2-(2-hydroxy-1-phenethyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII-7)
将化合物VI(2.81g,7.29mmol)，L-苯甘氨醇(VII-4)(1g,7.29mmol)和三乙胺(2.03mL,14.58mmol)溶于乙腈(30mL)，加热回流反应6h左右，TLC(二氯甲烷：甲醇＝20:1)检测原料反应完毕，减压蒸干溶剂，加入甲醇(10mL)冰浴冷却至10℃以下，搅拌30min，有固体析出，抽滤，得白色固体1.65g，产率：55.3％，m.p.154.2～155.8℃。Compound VI (2.81g, 7.29mmol), L-phenylglycinol (VII-4) (1g, 7.29mmol) and triethylamine (2.03mL, 14.58mmol) were dissolved in acetonitrile (30mL), heated to reflux for 6h Around, TLC (dichloromethane: methanol = 20:1) detects the completion of the reaction of the raw materials, evaporates the solvent under reduced pressure, adds methanol (10mL) in an ice bath to cool to below 10°C, stirs for 30min, a solid precipitates, suction filtration, white Solid 1.65g, yield: 55.3%, mp 154.2 ~ 155.8 ℃.
1H-NMR(300MHz,DMSO-d 6)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, CHCH 2OH),5.16-5.14(m,1H,CHCH 2 OH),4.75(d,J＝18.7Hz,1H, CH 2 N),4.43(d,J＝18.5Hz,1H, CH 2 N),4.06-3.95(m,2H,HO CH 2 CH),3.48(s,3H,SO 2 CH 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.13-9.12 (m, 1H, ArH), 8.56-8.53 (m, 3H, ArH), 7.81 (d, J = 7.8 Hz, 1H, ArH), 7.34-7.26 (m, 5H, ArH), 5.41-5.37 (m, 1H, CH CH 2 OH), 5.16-5.14 (m, 1H, CHCH 2 OH ), 4.75 (d, J = 18.7 Hz, 1H, CH 2 N), 4.43 (d, J = 18.5 Hz, 1H, CH 2 N), 4.06-3.95 (m, 2H, HO CH 2 CH), 3.48 (s, 3H, SO 2 CH 3 ).
(S)-2-(2-羟基-1-苯乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-22)的合成(S)-2-(2-Hydroxy-1-phenethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindoline- Synthesis of 1-ketone (I-22)
将化合物VIII-7(400mg,0.98mmol)和4-氨基四氢吡喃(IX-1)(198mg,1.96mmol)溶于仲丁醇(5mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝20：1)检测原料VIII-7反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体260mg，产率： 61.6％，m.p.143.9～145.6℃。Compound VIII-7 (400mg, 0.98mmol) and 4-aminotetrahydropyran (IX-1) (198mg, 1.96mmol) were dissolved in sec-butanol (5mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol = 20:1) detects that the raw material VIII-7 is completely reacted. The reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, and column chromatography (eluent: dichloromethane: methanol = 200:1) ~50:1) Separation gave 260 mg of white solid, yield: 61.6%, mp 143.9-145.6°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.17-5.13(m,1H, HOCH 2CH),4.71(d,J＝18.2Hz,1H,CONC H 2 ),4.40(d,J＝18.1Hz,1H,CONC H 2 ),4.09-3.98(m,3H,HO CH 2 CH,HN CH),3.91-3.88(m,2H,CH 2 CH 2 O CH 2 CH 2),3.44-3.42(m,2H,CH 2 CH 2 O CH 2 CH 2),1.91-1.85(m,2H,CH 2 CH 2 CH CH 2 CH 2),1.57-1.53(m,2H,CH 2 CH 2 CH CH 2 CH 2). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 8.42-8.31 (m, 3H, ArH), 7.71 (d, J=8.0 Hz, 1H, ArH), 7.35-7.24 (m, 6H, ArH), 5.42-5.34(m, 1H, HOCH 2 CH ), 5.17-5.13(m, 1H, HO CH 2 CH), 4.71(d, J=18.2Hz, 1H, CONC H 2 ), 4.40(d, J = 18.1 Hz, 1H, CONC H 2 ), 4.09-3.98 (m, 3H, HO CH 2 CH, HN CH ), 3.91-3.88 (m, 2H, CH 2 CH 2 O CH 2 CH 2 ), 3.44- 3.42(m,2H,CH 2 CH 2 O CH 2 CH 2 ),1.91-1.85(m,2H,CH 2 CH 2 CH CH 2 CH 2 ),1.57-1.53(m,2H,CH 2 CH 2 CH CH 2 CH 2 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75MHz, DMSO-d 6 ) 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 C 25H 27N 4O 3:431.2083；Found:431.2073. HRMS(ESI): m/z[M+H] + .Calcd for C 25 H 27 N 4 O 3 :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. 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.
实施例23Example 23
(S)-2-(2-羟基-1-苯乙基)-6-(2-((1-甲基-1H-吡唑-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-23)的合成(S)-2-(2-Hydroxy-1-phenethyl)-6-(2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)isoindole Synthesis of Pyridin-1-one (I-23)
将化合物VIII-7(400mg,0.98mmol)和1-甲基-1H-吡唑-4-胺(IX-3)(190mg,1.96mmol)溶于仲丁醇(5mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝20：1)检测原料VIII-7反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得白色固体206mg，产率：49.3％，m.p.227.9～229.7℃。Compound VIII-7 (400 mg, 0.98 mmol) and 1-methyl-1H-pyrazole-4-amine (IX-3) (190 mg, 1.96 mmol) were dissolved in sec-butanol (5 mL), and the tube was heated to 125 The reaction was carried out at ℃ for about 12h. After TLC (dichloromethane: methanol = 20:1), the raw material VIII-7 was detected to be complete. The reaction solution was cooled to room temperature, the solvent was evaporated under reduced pressure, and column chromatography (eluent: dichloromethane : Methanol = 200: 1 to 50: 1) Separation to obtain 206 mg of white solid, yield: 49.3%, mp 227.9 to 229.7 °C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.18-5.14(m,1H, HOCH 2),4.73(d,J＝18.2Hz,1H,CON CH 2 ),4.42(d,J＝18.4Hz,1H,CON CH 2 ),4.06-3.97(m,2H,HO CH 2 CH),3.83(s,3H,N CH 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.58 (s, 1H, HN), 8.52-8.51 (m, 1H, ArH), 8.46 (s, 1H, ArH), 8.38 (d, J = 8.4 Hz, 1H, ArH), 7.91 (s, 1H, ArH), 7.76 (d, J = 8.1 Hz, 1H, ArH), 7.57 (s, 1H, ArH), 7.41 (d, J = 5.1 Hz ,1H,ArH),7.36-7.31(m,5H,ArH),5.43-5.38(m,1H,HOCH 2 CH ),5.18-5.14(m,1H, HO CH 2 ),4.73(d,J=18.2 Hz, 1H, CON CH 2 ), 4.42 (d, J = 18.4 Hz, 1H, CON CH 2 ), 4.06-3.97 (m, 2H, HO CH 2 CH), 3.83 (s, 3H, N CH 3 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 24H 23N 6O 2:427.1882；Found:427.1871. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 23 N 6 O 2 : 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. IR (cm -1 ): 3409.61, 3261.17, 2292.35, 1167.93, 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.
实施例24Example 24
(S)-2-(2-羟基-1-苯乙基)-6-(2-((1-甲基-1H-吡唑-5-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-24)的合成(S)-2-(2-Hydroxy-1-phenethyl)-6-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)isoindole Synthesis of Pyridin-1-one (I-24)
(S)-2-(2-((叔丁基二甲基硅基)氧基)-1-苯乙基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(X-2)的合成(S)-2-(2-((tert-butyldimethylsilyl)oxy)-1-phenethyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindole Synthesis of Pyridin-1-one (X-2)
将化合物VIII-7(400mg,0.98mmol)和咪唑(133mg,1.96mmol)溶于二氯甲烷(4mL)，加入TBSCl(295mg,1.96mmol)，于35℃搅拌反应8h左右，待TLC(二氯甲烷:甲醇＝30：1)检测原料VIII-7反应完后，用饱和碳酸氢钠(4mL)洗涤，再用饱和氯化钠溶液(4mL)洗涤，无水硫酸钠干燥，过滤，减压蒸干溶剂，加入石油醚(4mL)打浆，过滤，得白色固体480mg，收率93.6％，m.p.70.2～72.1℃。Compound VIII-7 (400mg, 0.98mmol) and imidazole (133mg, 1.96mmol) were dissolved in dichloromethane (4mL), TBSCl (295mg, 1.96mmol) was added, and the reaction was stirred at 35 ℃ for about 8h, until TLC (dichloromethane) Methane: methanol = 30:1) After the detection of the raw material VIII-7 is completed, it is washed with saturated sodium bicarbonate (4mL), then with saturated sodium chloride solution (4mL), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure Dry solvent, add petroleum ether (4mL) to beat, and filter to obtain 480mg white solid, yield 93.6%, mp70.2-72.1℃.
1H-NMR(300MHz,DMSO-d 6))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,HOCH 2 CH),4.69(d,J＝18.5Hz,1H,CON CH 2 ),4.46(d,J＝18.4Hz,1H,CON CH 2 ),4.26-4.14(m,2H,O CH 2 ),3.49(s,3H,S CH 3 ),0.73(S,9H,C( CH 3) 3 ),0.01(d,J＝4.7Hz,6H,Si (CH 3) 2) . 1 H-NMR (300 MHz, DMSO-d 6 )) δ (ppm): 9.13 (d, J=5.4 Hz, 1H, ArH), 8.58-8.52 (m, 3H, ArH), 7.83 (d, J=7.92 Hz,1H,ArH),7.38-7.30(m.5H,ArH),5.45-5.41(m,1H,HOCH 2 CH ),4.69(d,J=18.5Hz,1H,CON CH 2 ),4.46(d , J = 18.4 Hz, 1H, CON CH 2 ), 4.26-4.14 (m, 2H, O CH 2 ), 3.49 (s, 3H, S CH 3 ), 0.73 (S, 9H, C ( CH 3 ) 3 ) , 0.01 (d, J = 4.7 Hz, 6H, Si (CH 3 ) 2 ) .
(S)-2-(2-羟基-1-苯乙基)-6-(2-((1-甲基-1H-吡唑-5-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-24)的合成(S)-2-(2-Hydroxy-1-phenethyl)-6-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)isoindole Synthesis of Pyridin-1-one (I-24)
将1-甲基-1H-吡唑-5-胺(IX-2)(74mg,0.76mmol)溶于无水四氢呋喃(3mL)，冷却至-78℃，加入1N LiHMDS(0.76mL,0.76mmol)，搅拌30min，将化合物X-2(200mg,0.38mmol)溶于无水四氢呋喃(3mL)，于-78℃下缓慢滴加入上述反应液，-78℃反应1h左右，TLC(二氯甲烷：甲醇＝20：1)检测原料X-2反应完全，减压蒸干溶剂，加入乙酸乙酯(5mL)溶解，用HCl的乙酸乙酯溶液调节pH＝2～3，继续搅拌反应3h左右，待TLC(二氯甲烷：甲醇＝15：1)检测中间体反应完全，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～30：1)分离，得金黄色固体目标化合物盐酸盐55mg，产率：31.3％，m.p.202.3～204.1℃。Dissolve 1-methyl-1H-pyrazole-5-amine (IX-2) (74mg, 0.76mmol) in anhydrous tetrahydrofuran (3mL), cool to -78°C, and add 1N LiHMDS (0.76mL, 0.76mmol) , Stir for 30min, dissolve compound X-2 (200mg, 0.38mmol) in anhydrous tetrahydrofuran (3mL), slowly add the above reaction solution dropwise at -78℃, react at -78℃ for about 1h, TLC (dichloromethane: methanol =20:1) Test the raw material X-2 for complete reaction, evaporate the solvent under reduced pressure, add ethyl acetate (5mL) to dissolve, adjust the pH=2~3 with ethyl acetate solution of HCl, continue to stir the reaction for about 3h, wait for TLC (Dichloromethane: methanol = 15:1) The intermediate reaction was detected to be complete, the solvent was evaporated under reduced pressure, and column chromatography (eluent: dichloromethane: methanol = 200: 1 to 30: 1) was separated to obtain golden yellow. The solid target compound hydrochloride 55 mg, yield: 31.3%, mp 202.3-204.1°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.22-5.18(m,1H, HOCH 2),4.78(d,J＝18.3Hz,1H, CH 2 NCO),4.47(d,J＝18.4 Hz,1H, CH 2 NCO),4.12-4.03(m,2H,HO CH 2 CH),3.77(s,3H,N CH 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) 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, HOCH 2 CH ), 5.22-5.18 (m, 1H, HO CH 2 ), 4.78 (d, J = 18.3 Hz, 1H, CH 2 NCO), 4.47 (d, J = 18.4 Hz, 1H, CH 2 NCO), 4.12-4.03 (m, 2H, HO CH 2 CH), 3.77 (s, 3H, N CH 3 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 24H 23N 6O 2:427.1882；Found:427.1867. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 23 N 6 O 2 : 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. IR (cm -1 ): 3396.69, 3323.24, 2923.20, 1665.59, 1558.09, 1449.72, 1407.88, 1307.11, 1269.59, 1203.11, 1062.83, 1042.64, 820.99, 768.85, 700.35, 613.30.
实施例25Example 25
(S)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-25)的合成(S)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl ) Synthesis of Isoindoline-1-one (I-25)
(S)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-(甲磺酰基)嘧啶-4-基)异吲哚啉-1-酮(VIII-8)的合成(S)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-(methylsulfonyl)pyrimidin-4-yl)isoindolin-1-one (VIII -8) Synthesis
将化合物VI(2,25g,5.83mmol)，(S)-2-氨基-2-(3-氯苯基)乙基-1-醇(VII-5)(1g,5,83mmol)和三乙胺(1.6mL,11.65mmol)溶于乙腈(30mL)，加热回流反应6h左右，待TLC(二氯甲烷：甲醇＝20:1)检测原料反应完全，减压蒸干溶剂，加入甲醇(10mL)冰浴冷却至10℃以下，搅拌30min，有固体析出，抽滤，得白色固体1.72g，产率：66.5％，m.p.71.8～73.5℃。Compound VI (2,25 g, 5.83 mmol), (S)-2-amino-2-(3-chlorophenyl)ethyl-1-ol (VII-5) (1 g, 5,83 mmol) and triethyl The amine (1.6mL, 11.65mmol) was dissolved in acetonitrile (30mL), heated to reflux for about 6h. After TLC (dichloromethane: methanol = 20:1) detected the raw material reaction was complete, the solvent was evaporated under reduced pressure, and methanol (10mL) was added The ice bath was cooled to below 10°C and stirred for 30 min. A solid precipitated and was filtered with suction to obtain 1.72 g of white solid. Yield: 66.5%, mp 71.8-73.5°C.
1H-NMR(300MHz,DMSO-d 6)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, CHCH 2OH),5.23-5.21(m,1H,CHCH 2 OH),4.78(d,J＝19.1Hz,1H, CH 2 N),4.51(d,J＝18.9Hz,1H, CH 2 N),4.08-4.00(m,2H,HO CH 2 CH),3.51(s,3H,S CH 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.16-9.14 (m, 1H, ArH), 8.60-8.55 (m, 3H, ArH), 7.84 (d, J = 8.8 Hz, 1H, ArH), 7.44-7.34 (m, 4H, ArH), 5.41-5.38 (m, 1H, CH CH 2 OH), 5.23-5.21 (m, 1H, CHCH 2 OH ), 4.78 (d, J = 19.1 Hz, 1H, CH 2 N), 4.51 (d, J = 18.9 Hz, 1H, CH 2 N), 4.08-4.00 (m, 2H, HO CH 2 CH), 3.51 (s, 3H, S CH 3 ).
(S)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-25)的合成(S)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl ) Synthesis of Isoindoline-1-one (I-25)
将化合物VIII-8(400mg,0.90mmol)和4-氨基四氢吡喃(IX-1)(182mg,1.80mmol)溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-8反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200:1～50:1)分离，得淡黄色固体220mg，产率：52.6％，m.p.86.2～87.9℃。Compound VIII-8 (400mg, 0.90mmol) and 4-aminotetrahydropyran (IX-1) (182mg, 1.80mmol) were dissolved in sec-butanol (3mL), sealed tube was heated to 125 ℃ for about 12h, wait TLC (dichloromethane: methanol = 25:1) detects that the raw material VIII-8 is completely reacted. The reaction solution is cooled to room temperature, the solvent is evaporated under reduced pressure, and column chromatography (eluent: dichloromethane: methanol = 200:1) ~50:1) Separation gave 220 mg of light yellow solid, yield: 52.6%, mp 86.2-87.9°C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.24-5.23(m,1H, HOCH 2CH),4.74(d,J＝18.3Hz,1H,CON CH 2 ),4.46(d,J＝18.0Hz,1H,CON CH 2 ),4.05-4.03(m,3H,HO CH 2 ,NH CH),3.92-3.89(m,2H,O(C H 2) 2),3.46-3.43(m,2H,O(C H 2) 2),1.92-1.88(m,2H,NHCH( CH 2) 2),1.62-1.51(m,2H,NHCH( CH 2) 2). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 8.44 (s, 1H, ArH), 8.40-8.39 (m, 1H, ArH), 8.34 (d, J=7.9 Hz, 1H, ArH) , 7.73 (d, J = 7.9 Hz, 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,HOCH 2 CH ),5.24-5.23(m,1H, HO CH 2 CH),4.74(d,J=18.3Hz,1H,CON CH 2 ),4.46(d,J=18.0Hz,1H, CON CH 2 ), 4.05-4.03(m,3H,HO CH 2 ,NH CH ),3.92-3.89(m,2H,O(C H 2 ) 2 ),3.46-3.43(m,2H,O(C H 2 ) 2 ), 1.92-1.88(m, 2H, NHCH( CH 2 ) 2 ), 1.62-1.51(m, 2H, NHCH( CH 2 ) 2 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 25H 26ClN 4O 3:465.1693；Found:465.1681. HRMS(ESI): m/z[M+H] + .Calcd for C 25 H 26 ClN 4 O 3 : 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. 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.
实施例26Example 26
(S)-2-(1-(3-氯苯基)-2-羟基乙基)-6-(2-((1-甲基-1H-吡唑-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-26)的合成(S)-2-(1-(3-chlorophenyl)-2-hydroxyethyl)-6-(2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidine-4 -Yl)isoindolin-1-one (I-26)
以化合物VIII-8(400mg,0.90mmol)和1-甲基-1H-吡唑-4-胺(IX-3)(175mg,1.80mmol)为原料，溶于仲丁醇(3mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-8反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200:1～50:1)分离，得金黄色固体126mg，产率：30.4％，m.p.207.8～209.6℃。Using compound VIII-8 (400mg, 0.90mmol) and 1-methyl-1H-pyrazole-4-amine (IX-3) (175mg, 1.80mmol) as raw materials, dissolved in sec-butanol (3mL), sealed tube Heat to 125°C for about 12 hours, wait for TLC (dichloromethane: methanol = 25:1) to detect the completion of the reaction of the raw material VIII-8, cool the reaction solution to room temperature, evaporate the solvent under reduced pressure, column chromatography (eluent: Dichloromethane: methanol = 200:1 ~ 50:1) separated to obtain 126 mg of golden yellow solid, yield: 30.4%, mp 207.8 ~ 209.6 °C.
1H-NMR(300MHz,DMSO-d 6)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,HOCH 2 CH),5.24-5.20(m,1H, HOCH 2CH),4.74(d,J＝17.9Hz,1H,CON CH 2 ),4.48(d,J＝18.0Hz,1H,CON CH 2 ),4.07-4.00(m,2H,HO CH 2 ),3.83(s,3H,N CH 3 ). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.58 (s, 1H, NH), 8.53-8.51 (m, 1H, ArH), 8.47 (s, 1H, ArH), 8.39 (d, J = 8.4 Hz, 1H, ArH), 7.92 (s, 1H, ArH), 7.77 (d, J = 8.0 Hz, 1H, ArH), 7.58 (brs, 1H, ArH), 7.44-7.32 (m, 5H, ArH), 5.41-5.37 (m, 1H, HOCH 2 CH ), 5.24-5.20 (m, 1H, HO CH 2 CH), 4.74 (d, J = 17.9 Hz, 1H, CON CH 2 ), 4.48 (d, J = 18.0 Hz, 1H, CON CH 2 ), 4.07-4.00 (m, 2H, HO CH 2 ), 3.83 (s, 3H, N CH 3 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) 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 C 24H 22ClN 6O 2:461.1493；Found:461.1482. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 22 ClN 6 O 2 : 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. 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.
实施例27Example 27
2-(3-氯苄基)-6-(2-((4-氟苯基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-27)的合成Synthesis of 2-(3-chlorobenzyl)-6-(2-((4-fluorophenyl)amino)pyrimidin-4-yl)isoindolin-1-one (I-27)
将化合物VIII-1(200mg,0.48mmol)和对氟苯胺IX-13(81mg,0.73mmol)溶于仲丁醇(5mL)，封管加热至125℃反应12h左右，待TLC(二氯甲烷：甲醇＝25：1)检测原料VIII-1反应完全，将反应液冷却至室温，减压蒸干溶剂，柱层析(洗脱剂：二氯甲烷：甲醇＝200:1～50:1)分离，得灰色固体98mg，产率：45.9％，196.2～197.3℃。Compound VIII-1 (200mg, 0.48mmol) and p-fluoroaniline IX-13 (81mg, 0.73mmol) were dissolved in sec-butanol (5mL), sealed tube was heated to 125 ℃ for about 12h, TLC (dichloromethane: Methanol = 25:1) The reaction of the raw material VIII-1 was detected to be complete. The reaction solution was cooled to room temperature, the solvent was evaporated under reduced pressure, and separated by column chromatography (eluent: dichloromethane: methanol = 200:1 to 50:1). To obtain 98 mg of gray solid, yield: 45.9%, 196.2 ~ 197.3 ℃.
1H-NMR(300MHz,DMSO-d 6)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,CH 2). 1 H-NMR (300 MHz, DMSO-d 6 ) δ (ppm): 9.76 (s, 1H, NH), 8.58 (d, J=5.2 Hz, 1H, ArH), 8.52 (s, 1H, ArH), 8.40 (dd, J = 8.1, 1.5 Hz, 1H, ArH), 7.83 (dd, J = 8.8, 4.8 Hz, 2H, ArH), 7.74 (d, J = 8.0 Hz, 1H, ArH), 7.54 (d, J = 5.2 Hz, 1H, ArH), 7.19-7.13 (m, 3H, ArH), 7.28 (d, J = 6.4 Hz, 1H, ArH), 7.16 (t, J = 8.9 Hz, 2H, ArH), 4.78 ( s, 2H, CH2), 4.50 (s, 2H, CH 2 ).
13C-NMR(75MHz,DMSO-d 6)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. 13 C-NMR (75 MHz, DMSO-d 6 ) δ (ppm): 167.05, 162.88, 161.41, 159.71 (d, J = 66.2 Hz), 159.22, 144.36, 139.97, 136.79 (d, J = 8.5 Hz), 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 C 25H 19ClFN 4O:445.1226；Found:445.1235. HRMS(ESI): m/z[M+H] + .Calcd for C 25 H 19 ClFN 4 O: 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. IR (cm -1 ): 3465.54, 3265.05, 1703.97, 1570.27, 1537.36, 1507.27, 1463.32, 1453.48, 1430.72, 1303.10, 1119.33, 829.97, 801.87, 764.94, 711.27, 612.21, 548.43.
实施例28Example 28
2-((6-甲基吡啶-2-基)甲基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-28)的合成2-((6-Methylpyridin-2-yl)methyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindoline- Synthesis of 1-ketone (I-28)
2-(溴甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊烷-2-基)苯甲酸甲酯(XVII)的合成Methyl 2-(bromomethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)benzoate (XVII) synthesis
将化合物III(1g,3.6mmol)和NBS(765mg,4.3mmol)溶于四氯化碳(15mL)，加入过氧化苯甲酰(12mg,0.036mmol)，加热至80℃反应4h左右，TLC(石油醚：乙酸乙酯＝20:1)检测原料III反应完全。冷却至室温，滤掉不溶物，滤液浓缩得红色油状物，不经处理直接投下一步。Compound III (1g, 3.6mmol) and NBS (765mg, 4.3mmol) were dissolved in carbon tetrachloride (15mL), added benzoyl peroxide (12mg, 0.036mmol), heated to 80 ℃ for about 4h, TLC ( Petroleum ether: ethyl acetate=20:1) The reaction of raw material III was detected to be complete. After cooling to room temperature, the insoluble matter was filtered off, and the filtrate was concentrated to obtain a red oil, which was directly sent to the next step without treatment.
2-((6-甲基吡啶-2-基)甲基)-6-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊烷-2-基)异吲哚啉-1-酮(XVIII)的合成2-((6-methylpyridin-2-yl)methyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2- )Isoindoline-1-one (XVIII) Synthesis
将6-甲基-2-氨甲基吡啶(VII-5)(194mg,1.8mmol)和三乙胺(182mg,1.8mmol)溶于乙腈(10mL)，滴入化合物XVII(714mg,2mmol)，加热至80℃反应10h左右。TLC(二氯甲烷：甲醇＝20：1)检测原料XVII反应完全，减压蒸干溶剂，乙酸乙酯(15mL)溶解，水洗，干燥，过滤，滤液减压蒸除溶剂，残留物柱层析(洗脱剂：二氯甲烷：甲醇＝75：1)分离，得深绿色油状物157mg，产率：23.9％。6-Methyl-2-aminomethylpyridine (VII-5) (194 mg, 1.8 mmol) and triethylamine (182 mg, 1.8 mmol) were dissolved in acetonitrile (10 mL), and compound XVII (714 mg, 2 mmol) was added dropwise. Heated to 80 ℃ for about 10h. TLC (dichloromethane: methanol = 20:1) detected that the reaction of the raw material XVII was complete, the solvent was evaporated under reduced pressure, ethyl acetate (15 mL) was dissolved, washed with water, dried, filtered, the filtrate was distilled off the solvent under reduced pressure, and the residue was subjected to column chromatography (Eluent: dichloromethane: methanol = 75:1) isolated to obtain 157 mg of dark green oil, yield: 23.9%.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.49(s,2H,CH 2),2.60(s,3H,CH 3),1.39(s,12H,CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.40 (s, 1H, ArH), 7.99 (d, J=7.6 Hz, 1H ArH), 7.57 (t, J=7.7 Hz, 1H ArH), 7.45 (d, J = 7.6 Hz, 1H ArH), 7.14 (d, J = 7.8 Hz, 1H ArH), 7.10 (d, J = 7.6 Hz, 1H ArH), 4.97 (s, 2H, CH 2 ), 4.49 (s, 2H, CH 2 ), 2.60 (s, 3H, CH 3 ), 1.39 (s, 12H, CH 3 ).
6-(2-氯嘧啶-4-基)-2-((6-甲基吡啶-2-基)甲基)异吲哚啉-1-酮(XIX)的合成Synthesis of 6-(2-chloropyrimidin-4-yl)-2-((6-methylpyridin-2-yl)methyl)isoindolin-1-one (XIX)
将XVIII(500mg,1.4mmol)，2,4-二氯嘧啶(XX)(207mg,1.4mmol),四三苯基膦钯(162mg,0.14mmol)和磷酸钾(890mg,4.2mmol)溶于二氧六环/水混合溶液(20/5mL)，氮气保护下加热至80℃反应12h左右。TLC(二氯甲烷：甲醇＝25：1)检测原料XVIII反应完全，冷却至室温，过滤，滤液减压浓缩，残留物柱层析(洗脱剂：二氯甲烷：甲醇＝50：1)分离，得黄色固体272mg，产率：55.4％，m.p.208.3～209.2℃。Dissolve XVIII (500 mg, 1.4 mmol), 2,4-dichloropyrimidine (XX) (207 mg, 1.4 mmol), tetratriphenylphosphine palladium (162 mg, 0.14 mmol) and potassium phosphate (890 mg, 4.2 mmol) in two Oxygen hexacyclic/water mixed solution (20/5mL), heated to 80℃ under nitrogen protection for about 12h. TLC (dichloromethane: methanol = 25:1) detected that the reaction of the raw material XVIII was complete, cooled to room temperature, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by column chromatography (eluent: dichloromethane: methanol = 50:1). To obtain 272mg of yellow solid, yield: 55.4%, mp 208.3 ~ 209.2 ℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.60(s,2H,CH 2),2.60(s,3H,CH 3). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.74-8.72 (m, 1H, ArH), 8.54 (s, 1H, ArH), 8.47 (d, J=7.9 Hz, 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, CH 2 ), 4.60 (s, 2H, CH 2 ), 2.60 (s, 3H, CH 3 ).
2-((6-甲基吡啶-2-基)甲基)-6-(2-((四氢-2H-吡喃-4-基)氨基)嘧啶-4-基)异吲哚啉-1-酮(I-28)的合成2-((6-Methylpyridin-2-yl)methyl)-6-(2-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)isoindoline- Synthesis of 1-ketone (I-28)
将XIX(50mg,0.14mmol)，4-氨基四氢吡喃(IX-1)(14mg,0.14mmol)，Pd(OAc) 2(3mg,0.014mmol)，4,5-双二苯基膦-9,9-二甲基氧杂蒽(35mg,0.06mmol)和Cs 2CO 3(68mg,0.21mmol)溶于二氧六环(3mL)，氮气保护下加热至100℃反应12h左右，TLC(二氯甲烷：甲醇＝20：1)检测原料XIX反应完全，反应液冷却至室温，加入乙酸乙酯稀释，分别用水洗(2mL)，饱和食盐水洗(2mL)，有机层用无水硫酸钠干燥，抽滤，滤液减压蒸干溶剂，残留物柱层析(洗脱剂：二氯甲烷：甲醇＝200：1～50：1)分离，得淡黄色固体24mg，产率：41.3％，m.p.186.1～187.2℃。 Combine XIX (50 mg, 0.14 mmol), 4-aminotetrahydropyran (IX-1) (14 mg, 0.14 mmol), Pd(OAc) 2 (3 mg, 0.014 mmol), 4,5-bisdiphenylphosphine- 9,9-Dimethylxanthene (35mg, 0.06mmol) and Cs 2 CO 3 (68mg, 0.21mmol) were dissolved in dioxane (3mL), heated to 100℃ under nitrogen protection for about 12h, TLC ( Dichloromethane: methanol = 20:1) The reaction of the raw material XIX was detected to be complete. The reaction solution was cooled to room temperature, diluted with ethyl acetate, washed with water (2 mL), saturated brine (2 mL), and the organic layer was dried over anhydrous sodium sulfate. , Suction filtration, the filtrate was evaporated to dryness under reduced pressure, the residue was separated by column chromatography (eluent: dichloromethane: methanol = 200: 1 ~ 50: 1), to give a light yellow solid 24mg, yield: 41.3%, mp186 .1～187.2℃.
1H-NMR(300MHz,CDCl 3)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,CH 2),4.55(s,2H,CH 2),4.29-4.10(m,1H,NHCH),4.04-4.00(m,2H,OCH 2),3.63-3.56(m,2H,OCH 2),2.58(s,3H,CH 3),2.13-2.08(m,2H,NHCHCH 2),1.67-1.57(m,2H,NHCHCH 2). 1 H-NMR (300 MHz, CDCl 3 ) δ (ppm): 8.48 (s, 1H, ArH), 8.36 (d, J=5.3 Hz, 1H, ArH), 8.27 (d, J=7.7 Hz, 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,CH 2 ),4.55(s,2H,CH 2 ),4.29-4.10(m,1H,NHCH),4.04-4.00(m,2H,OCH 2 ),3.63-3.56(m,2H,OCH 2 ), 2.58 (s, 3H, CH 3 ), 2.13-2.08 (m, 2H, NHCHCH 2 ), 1.67-1.57 (m, 2H, NHCHCH 2 ).
13C-NMR(75MHz,CDCl 3)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. 13 C-NMR (75 MHz, CDCl 3 ) 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 C 24H 26N 5O 2:416.2087；Found:416.2081. HRMS(ESI): m/z[M+H] + .Calcd for C 24 H 26 N 5 O 2 :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. 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.
实施例29Example 29
上述制备所得部分化合物的药理学实验及结果如下：The pharmacological experiments and results of some of the compounds prepared above are as follows:
一、ERK2酶抑制活性实验1. ERK2 enzyme inhibitory activity experiment
①实验方法① Experimental method
将所有化合物用DMSO溶解，配置10mM母液，然后把化合物加到筛选体系中，化合物的检测浓度范围是0.05nM-1μM，按照3倍梯度进行稀释，每个浓度做两个复孔。实验结果换算成活性百分率，描绘量效曲线，用GRAPHPAD PEISM 5非线性回归计算得到抑制IC 50值。 Dissolve all compounds in DMSO, prepare 10mM mother liquor, and then add the compound to the screening system. The detection concentration of the compound is in the range of 0.05nM-1μM. Dilute according to a 3-fold gradient, and make two replicate wells for each concentration. The experimental results were converted into percentages of activity, and the dose-effect curve was drawn. The inhibition IC 50 value was calculated by GRAPHPAD PEISM 5 nonlinear regression calculation.
酶反应体系组成如下：20mM Hepes(pH 7.5),10mM MgCl 2,1mM EGTA,0.02％Brij35,0.02mg/mL BSA,0.1mM Na 3VO 4,2mM DTT,10μM ATP，激酶，激酶底物；同时加入不同浓度的待筛选化合物，组成50μl的反应体系，在室温下反应2h，后用荧光素酶的方法检测体系内的ADP含量，再反应5min后，在MD-SpectraMax M5多功能酶标仪上检测化学发光信号，化学发光信号值强度与酶活性抑制成正比。检测到的化学发光信号值，代入公式： The composition of the enzyme reaction system is as follows: 20 mM Hepes (pH 7.5), 10 mM MgCl 2 , 1 mM EGTA, 0.02% Brij35, 0.02 mg/mL BSA, 0.1 mM Na 3 VO 4 , 2 mM DTT, 10 μM ATP, kinase, kinase substrate; Add different concentrations of the compound to be screened to form a 50μl reaction system, react at room temperature for 2h, then use the luciferase method to detect the ADP content in the system, and then react for 5min, on the MD-SpectraMax M5 multi-functional microplate reader The chemiluminescence signal is detected, and the intensity of the chemiluminescence signal is proportional to the inhibition of enzyme activity. The detected chemiluminescence signal value is substituted into the formula:
酶活性百分率(％)＝{(Lu药物-Lu本底)/(Lu酶-Lu本底)}x100％Enzyme activity percentage (%) = {(Lu drug-Lu background)/(Lu enzyme-Lu background)} x100%
然后用Graphpad Prism 5软件处理，计算出化合物的IC 50值。 Then use Graphpad Prism 5 software to calculate the IC 50 value of the compound.
②试验结果：② Test result:
对本发明部分化合物进行体外ERK2激酶抑制活性的筛选，结果见表1。Some compounds of the present invention were screened for ERK2 kinase inhibitory activity in vitro. The results are shown in Table 1.
表1.部分受试化合物对ERK2激酶的IC 50值 Table 1. IC50 values of the test compound partially kinase ERK2 IC
Figure PCTCN2019104110-appb-000022
Figure PCTCN2019104110-appb-000022
表1结果显示，本发明化合物对ERK2激酶均有较好的抑制活性，其中化合物I-8和I-10的活性较好，对ERK2激酶的IC 50分别为0.7nM和1.2nM。同时分别对比化合物I-8和I-10的S构型和R构型的药理活性可知，S构型的化合物活性远远优于R构型化合物。 The results in Table 1 show that the compounds of the present invention have better inhibitory activity against ERK2 kinase, of which compounds I-8 and I-10 have better activities, and the IC 50 of ERK2 kinase is 0.7 nM and 1.2 nM, respectively. At the same time, comparing the pharmacological activities of S configuration and R configuration of compounds I-8 and I-10, it is known that the activity of S configuration compound is far superior to that of R configuration compound.
二、人肿瘤细胞增殖抑制作用实验2. Experiment of human tumor cell proliferation inhibition
①实验方法① Experimental method
试验分组情况及药物浓度选择Trial grouping and drug concentration selection
所有化合物浓度选择：1nM,10nM,100nM,500nM,1000nM。All compound concentration options: 1nM, 10nM, 100nM, 500nM, 1000nM.
细胞培养条件(Cell Culture)Cell culture conditions (Cell Culture)
细胞传代培养10-15代，培养条件为含有青霉素(终浓度为100U/mL)、链霉素(终浓度为100μg/mL)、10％FBS的培养基(CT26、Colo-205、WM-266-4：RPMI-1640；HCT116、HT29：McCoy's 5a；SW626：L-15)，当 细胞融合至90％时，弃去旧培养基，用2mL PBS洗涤细胞2次，弃去PBS后加入2mL的0.25％(w/v)Trypsin-0.53mM EDTA混合消化液，置显微镜下观察，约30s，当细胞变圆后迅速加入2mL完全培养基终止消化，轻轻吹打，收集细胞。800rpm，4℃，离心5min，弃去上清，用完全培养基重悬细胞，分瓶培养，隔天换液。Cells were subcultured for 10-15 generations, and the culture conditions were medium containing penicillin (final concentration 100U/mL), streptomycin (final concentration 100μg/mL), 10% FBS (CT26, Colo-205, WM-266 -4: RPMI-1640; HCT116, HT29: McCoy's 5a; SW626: L-15), when the cells are fused to 90%, discard the old medium, wash the cells twice with 2mL PBS, add 2mL after discarding the PBS 0.25% (w/v) Trypsin-0.53 mM EDTA mixed digestion solution, observed under a microscope, about 30s, when the cells become round, quickly add 2mL of complete medium to stop the digestion, gently pipet to collect the cells. Centrifuge at 800 rpm, 4°C for 5 min, discard the supernatant, resuspend the cells in complete medium, culture in separate flasks, and change the medium the next day.
MTT试验MTT test
将对数生长期细胞以1×105cells/孔接种于96孔板，置于37℃，5％CO 2条件下培养，直至细胞90％融合后，用无血清的L-15培养基孵育2h使细胞同步化。随后，弃去上清液，分别加入含有各化合物(化合物加入后最终稀释浓度1nM,10nM,100nM,500nM,1000nM)的L-15培养基孵育72h，孵育结束前4h，每孔加入20μl MTT溶液(5mg/mL)。孵育结束后，弃去各孔上清液，每孔加入150μL DMSO，细胞振荡仪上振荡10min，待结晶物充分溶解后用酶标仪测定OD570。 Log growth phase cells were seeded in 96-well plates at 1×105cells/well and cultured at 37°C under 5% CO 2 until the cells were 90% confluent, and incubated with serum-free L-15 medium for 2h Cell synchronization. Subsequently, the supernatant was discarded, and L-15 medium containing each compound (final dilution concentration of 1 nM, 10 nM, 100 nM, 500 nM, 1000 nM after compound addition) was added for incubation for 72 h. 4 h before the end of incubation, 20 μl of MTT solution was added to each well (5mg/mL). After the incubation, the supernatant of each well was discarded, 150 μL of DMSO was added to each well, and shaken on a cell shaker for 10 min. After the crystals were fully dissolved, the OD570 was measured with a microplate reader.
增殖抑制活性百分率(％)＝(OD值给药-OD值本底)/(OD值对照-OD值本底)x100％Percentage of proliferation inhibitory activity (%) = (OD value administration-OD value background) / (OD value control-OD value background) x 100%
将药物浓度作为横坐标，各浓度对应的增殖抑制活性百分率为纵坐标，使用Graphpd Prism5做非线性回归，计算得各个化合物的IC 50值。 Taking the drug concentration as the abscissa, the percentage of proliferation inhibitory activity corresponding to each concentration is the ordinate, and non-linear regression was performed using Graphpd Prism5 to calculate the IC 50 value of each compound.
②试验结果：② Test result:
选取部分具有较好ERK2激酶抑制活性的化合物，测定其对含有RAS-RAF-MEK-ERK突变的肿瘤细胞系人结肠癌细胞Colo-205(BRAFV600E)、人黑色素瘤细胞WM-266-4(BRAFV600D)、人卵巢癌细胞SW-626(KRASG12V)和人结肠癌细胞HCT-116(KRASG13D)的抗增殖能力。实验结果显示(表2)，受试化合物对四种细胞系均表现出一定的增殖抑制能力，抑制活性与阳性药BVD-523相当。Some compounds with better ERK2 kinase inhibitory activity were selected and tested for human colon cancer cell line Colo-205 (BRAFV600E) and human melanoma cell WM-266-4 (BRAFV600D) containing RAS-RAF-MEK-ERK mutation ), anti-proliferative ability of human ovarian cancer cell SW-626 (KRASG12V) and human colon cancer cell HCT-116 (KRASG13D). The experimental results showed (Table 2) that the test compounds showed a certain degree of proliferation inhibitory ability on the four cell lines, and the inhibitory activity was comparable to that of the positive drug BVD-523.
表2部分化合物对四种ERK通路活化细胞系抗增殖作用Table 2 Anti-proliferative effects of some compounds on four ERK pathway activated cell lines
Figure PCTCN2019104110-appb-000023
Figure PCTCN2019104110-appb-000023
在此基础上，测定上述化合物对不含RAS-RAF-MEK-ERK突变的肿瘤细胞系人慢性髓系白血病细胞K562的增殖抑制能力(表3)。实验结果显示，受试化合物以及BVD-523对K562细胞增殖的抑制活性(IC 50均大于1μM)明显弱于上述4个含有RAS-RAF-MEK-ERK突变的肿瘤细胞系，抗增殖活性大大减弱，说明受试化合物对激酶具有一定的选择性，可以选择性抑制ERK激酶。 On this basis, the above compounds were tested for their ability to inhibit the proliferation of human chronic myeloid leukemia cell K562, a tumor cell line that does not contain the RAS-RAF-MEK-ERK mutation (Table 3). The experimental results show that the inhibitory activity of the test compound and BVD-523 on the proliferation of K562 cells (IC 50 is greater than 1 μM) is significantly weaker than the above four tumor cell lines containing the RAS-RAF-MEK-ERK mutation, and the anti-proliferative activity is greatly reduced , Indicating that the test compound has certain selectivity for kinase and can selectively inhibit ERK kinase.
表3部分化合物对ERK通路非活化细胞系抗增殖作用Table 3 Anti-proliferative effects of some compounds on ERK pathway non-activated cell lines
Figure PCTCN2019104110-appb-000024
Figure PCTCN2019104110-appb-000024
三、NCM-460细胞毒性实验3. NCM-460 cytotoxicity experiment
①实验方法① Experimental method
试验分组情况及药物浓度选择Trial grouping and drug concentration selection
所有化合物浓度选择：10nM,100nM,500nM,1000nM,10000nM。All compound concentration options: 10nM, 100nM, 500nM, 1000nM, 10000nM.
细胞培养条件(Cell Culture)Cell culture conditions (Cell Culture)
细胞传代培养，培养条件为含有青霉素(终浓度为100U/mL)、链霉素(终浓度为100μg/mL)、10％FBS的RPMI-1640培养基，当细胞融合至90％时，弃去旧培养基，用2mL PBS洗涤细胞2次，弃去PBS后加入2mL的0.25％(w/v)Trypsin-0.53mM EDTA混合消化液，置显微镜下观察，约30s，当细胞变圆后迅速加入2mL完全培养基终止消化，轻轻吹打，收集细胞。800rpm，4℃，离心5min，弃去上清，用完全培养基重悬细胞，分瓶培养，隔天换液。Subculture of cells, the culture conditions are RPMI-1640 medium containing penicillin (final concentration 100U/mL), streptomycin (final concentration 100μg/mL), 10% FBS, when the cells are confluent to 90%, discard Wash the cells twice with 2mL of PBS, discard the PBS, add 2mL of 0.25% (w/v) Trypsin-0.53mM EDTA mixed digestion solution, and observe under the microscope for about 30s. When the cells become round, add them quickly 2mL complete medium was used to stop the digestion, gently pipetting to collect the cells. Centrifuge at 800 rpm, 4°C for 5 min, discard the supernatant, resuspend the cells in complete medium, culture in separate flasks, and change the medium the next day.
MTT试验MTT test
将对数生长期细胞以1×10 5cells/孔接种于96孔板，置于37℃，5％CO 2条件下培养，直至细胞90％融合后，用无血清的L-15培养基孵育2h使细胞同步化。随后，弃去上清，分别加入含有各化合物(10nM,100nM,1000nM,5000nM,10000nM)的RPMI-1640培养基孵育72h，孵育结束前4h，每孔加入20μL MTT溶液(5mg/mL)。孵育结束后，弃去各孔上清液，每孔加入150μL DMSO，细胞振荡仪上振荡10min，待结晶物充分溶解后用酶标仪测定OD 570。 Log growth phase cells were seeded in 96-well plates at 1×10 5 cells/well and cultured at 37°C under 5% CO 2 until the cells were 90% confluent and incubated with serum-free L-15 medium Synchronize the cells for 2h. Subsequently, the supernatant was discarded, and the RPMI-1640 medium containing each compound (10 nM, 100 nM, 1000 nM, 5000 nM, 10000 nM) was added for incubation for 72 h, and 4 μL of MTT solution (5 mg/mL) was added to each well before incubation for 4 h. After the incubation, the supernatant of each well was discarded, 150 μL of DMSO was added to each well, and shaken on a cell shaker for 10 min. After the crystals were fully dissolved, the OD 570 was measured with a microplate reader.
②试验结果：② Test result:
表4部分化合物对NCM-460细胞的抗增殖作用Table 4 Anti-proliferative effects of some compounds on NCM-460 cells
Figure PCTCN2019104110-appb-000025
Figure PCTCN2019104110-appb-000025
我们选取了化合物I-8和I-10，利用NCM-460细胞系测定其对正常细胞的抗增殖能力(表4)。结果表明两种化合物对NCM-460细胞生长均表现出极弱的增殖抑制作用，IC 50值均＞10000nM。 We selected compounds I-8 and I-10, and measured their anti-proliferative ability against normal cells using NCM-460 cell line (Table 4). The results showed that both compounds NCM-460 cells were grown exhibits very weak inhibition of proliferation, IC 50 values were> 10000nM.
通过以上数据我们发现，化合物I-8和I-10对NCM-460正常细胞生长无明显毒性。From the above data, we found that compounds I-8 and I-10 have no obvious toxicity to the growth of NCM-460 normal cells.
四、HCT-116裸小鼠移植瘤抑制实验4. HCT-116 nude mice transplantation tumor inhibition experiment
①实验方法① Experimental method
试验动物分组情况：Grouping of test animals:
阴性对照组(模型组)：10只，腹腔给予含5％DMSO的生理盐水；Negative control group (model group): 10 rats, intraperitoneal administration of saline containing 5% DMSO;
I-8组：10只，给药剂量为50mg/kg；Group I-8: 10 animals, the dosage is 50mg/kg;
BVD-523组：10只，给药剂量为50mg/kg；BVD-523 group: 10 animals, the dosage is 50mg/kg;
给药方式：每日一次，灌胃给药。Mode of administration: once a day, intragastric administration.
移植瘤模型的建立与药物治疗：Establishment of transplanted tumor model and drug treatment:
HCT116细胞传代培养10-15代，培养条件为含有青霉素(终浓度为100U/mL)、链霉素(终浓度为100μg/mL)、10％FBS的1640培养基，当细胞融合至90％时，弃去旧培养基，用2mL PBS洗涤细胞2次，弃去PBS后加入2ml 0.25％胰蛋白酶-0.02％EDTA混合消化液，置显微镜下观察，约30s，当细胞变圆后迅速加入2ml完全培养基终止消化，轻轻吹打，收集细胞。800rpm，4℃，离心5min，弃去上清液，用完全培养基重悬细胞，分瓶培养，隔天换液。HCT116 cells were subcultured for 10-15 passages under 1640 medium containing penicillin (final concentration 100U/mL), streptomycin (final concentration 100μg/mL), 10% FBS, when the cells were confluent to 90% , Discard the old culture medium, wash the cells twice with 2mL of PBS, add 2ml of 0.25% trypsin-0.02% EDTA mixed digestion solution after discarding the PBS, observe under the microscope, about 30s, when the cells become round, quickly add 2ml completely The medium is terminated for digestion, gently pipetting to collect the cells. Centrifuge at 800 rpm, 4°C for 5 min, discard the supernatant, resuspend the cells in complete medium, culture in separate flasks, and change the fluid the next day.
取对数生长期的HCT116细胞，在无菌条件下用不含血清的培养基制备成5×10 6/mL细胞悬液，并选取健康的上述实验裸鼠30只，饲养一周，待体重约为20g，每只裸鼠左侧腋窝皮下注射0.2mL的细胞悬液，细胞浓度5×10 7/mL。裸鼠移植瘤用游标卡尺测量移植瘤直径，待肿瘤生长至100mm 3左右后将动物随机分组记为第0天，然后开始按照分组方式中的给药方法给药。使用测量肿瘤直径的方法，动态观察被试化合物抗肿瘤的效应。肿瘤直径的测量次数为每3天一次。给药体积各组不同。30天后，小鼠处死，手术剥取瘤块称重。肿瘤体积(tumor volume,TV)的计算公式为： Take HCT116 cells in the logarithmic growth phase, prepare a 5×10 6 /mL cell suspension with a serum-free medium under sterile conditions, and select 30 healthy nude mice from the above experiment and feed them for one week. For 20 g, each nude mouse was injected subcutaneously with 0.2 mL of cell suspension in the left armpit, with a cell concentration of 5×10 7 /mL. Transplanted tumors of nude mice were measured with a vernier caliper. After the tumors grew to about 100 mm 3 , the animals were randomly grouped and recorded as day 0, and then started to be administered according to the method of grouping. Using the method of measuring tumor diameter, the anti-tumor effect of the test compound was dynamically observed. The number of tumor diameter measurements is every 3 days. The administration volume is different in each group. After 30 days, the mice were sacrificed and the tumor mass was surgically stripped and weighed. The calculation formula of tumor volume (TV) is:
TV＝1/2×a×b 2 TV=1/2×a×b 2
其中a、b分别表示长宽。Where a and b represent length and width, respectively.
统计分析：Statistical Analysis:
数据组间统计学差异采用one-way ANOVA和Tukey’s检验，P值小于0.05认为有显著性差异。The statistical difference between the data groups was determined by one-way ANOVA and Tukey’s test. A P value of less than 0.05 was considered to be a significant difference.
②试验结果：② Test result:
我们选取化合物I-8，利用HCT-116肿瘤细胞的小鼠移植瘤模型评价化合物I-8的体内抗肿瘤活性，并与阳性药BVD-523进行了比较。We selected compound I-8 and evaluated the antitumor activity of compound I-8 in vivo using a mouse transplanted tumor model of HCT-116 tumor cells, and compared it with the positive drug BVD-523.
如图1所示，以50mg/kg/day连续灌胃给药30天，化合物I-8与BVD-523均能够显著抑制HCT-116裸鼠移植瘤体积及重量的增加(p<0.05)，I-8的肿瘤抑制率为71％，BVD-523的肿瘤抑制率为54％，两者均不会对小鼠体重产生影响。实验结果显示，I-8在灌胃给药50mg/kg剂量下的抗肿瘤作用优于阳性药BVD-523。As shown in Figure 1, compound I-8 and BVD-523 were able to significantly inhibit the increase in the volume and weight of transplanted tumors of HCT-116 nude mice (p<0.05) after continuous oral administration at 50 mg/kg/day for 30 days. The tumor suppression rate of I-8 is 71%, and the tumor suppression rate of BVD-523 is 54%, neither of which will affect the body weight of mice. The experimental results show that the anti-tumor effect of I-8 at the dose of 50 mg/kg administered by gavage is superior to the positive drug BVD-523.

Claims (10)

  1. 一种如通式I所示的异吲哚啉酮类衍生物或其药学上可接受的盐：An isoindolinone derivative represented by general formula I or a pharmaceutically acceptable salt thereof:
    Figure PCTCN2019104110-appb-100001
    Figure PCTCN2019104110-appb-100001
    其中：among them:
    R 1选自
    Figure PCTCN2019104110-appb-100002
    Figure PCTCN2019104110-appb-100003
    其中，X选自CH 2、O、NH或CH-OH；R 4选自H或C1～C6的烷基，R 5选自H、CH 3、-CH 2OH、-CH 2CH 2OH、-CH 2CH 2CH 2OH或-CH 2CH 2CH 2CH 2OH；Y选自CH 2、O、NH或S；R 6选自F、Cl、Br、CH 3、NH 2或NHCOCH 3；     R 1 is selected from
    Figure PCTCN2019104110-appb-100002
    Figure PCTCN2019104110-appb-100003
    Wherein, X is selected from CH 2 , O, NH or CH-OH; R 4 is selected from H or C1-C6 alkyl, R 5 is selected from H, CH 3 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH; Y is selected from CH 2 , O, NH or S; R 6 is selected from F, Cl, Br, CH 3 , NH 2 or NHCOCH 3 ;
    R 2为H或-CH 2OH； R 2 is H or -CH 2 OH;
    R 3选自
    Figure PCTCN2019104110-appb-100004
    其中，R 7为H、F、Cl、Br或OCH 3，R 7为单取代、双取代或三取代；R 8为H或CH 3。     R 3 is selected from
    Figure PCTCN2019104110-appb-100004
    Wherein, R 7 is H, F, Cl, Br or OCH 3 , R 7 is mono-substituted, di-substituted or tri-substituted; R 8 is H or CH 3 .
  2. 根据权利要求1所述的异吲哚啉酮类衍生物I或其药学上可接受的盐，其特征在于：The isoindolinone derivative I according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that:
    R 1选自
    Figure PCTCN2019104110-appb-100005
    Figure PCTCN2019104110-appb-100006
    R 1 is selected from
    Figure PCTCN2019104110-appb-100005
    Figure PCTCN2019104110-appb-100006
  3. 根据权利要求1所述的异吲哚啉酮类衍生物I或其药学上可接受的盐，其特征在于：The isoindolinone derivative I according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that:
    R 3选自
    Figure PCTCN2019104110-appb-100007
    R 3 is selected from
    Figure PCTCN2019104110-appb-100007
  4. 根据权利要求1-3中任一所述的异吲哚啉酮类衍生物I或其药学上可接受的盐，其特征在于：R 2为-CH 2OH时，所述化合物I或其药学上可接受的盐包括手性异构体：
    Figure PCTCN2019104110-appb-100008
    Figure PCTCN2019104110-appb-100009
    The isoindolinone derivative I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein when R 2 is -CH 2 OH, the compound I or its pharmacy Acceptable salts include chiral isomers:
    Figure PCTCN2019104110-appb-100008
    Figure PCTCN2019104110-appb-100009
  5. 根据权利要求1所述的异吲哚啉酮类衍生物I或其药学上可接受的盐，其特征在于，所述异吲哚啉酮类衍生物选自I-1至I-28：The isoindolinone derivatives I or pharmaceutically acceptable salts thereof according to claim 1, wherein the isoindolinone derivatives are selected from I-1 to I-28:
    Figure PCTCN2019104110-appb-100010
    Figure PCTCN2019104110-appb-100010
    Figure PCTCN2019104110-appb-100011
    Figure PCTCN2019104110-appb-100011
  6. 根据权利要求1所述的异吲哚啉酮类衍生物I或其药学上可接受的盐，其特征在于，所述药学上可接受的盐为通式I化合物的酸加成盐，其中用于成盐的酸为：氯化氢、溴化氢、硫酸、碳酸、草酸、柠檬酸、琥珀酸、酒石酸、磷酸、乳酸、丙酮酸、乙酸、马来酸、甲磺酸、苯磺酸、对甲苯磺酸或阿魏酸。The isoindolinone derivatives I according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that the pharmaceutically acceptable salt is an acid addition salt of the compound of formula I, wherein Salt-forming acids are: 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-toluene Sulfonic acid or ferulic acid.
  7. 一种权利要求1所述异吲哚啉酮类衍生物的制备方法，其特征在于，由化合物VIII与IX经取代反应制备化合物I：A method for preparing isoindolinone derivatives according to claim 1, wherein compound I is prepared by substitution reaction of compound VIII and IX:
    Figure PCTCN2019104110-appb-100012
    Figure PCTCN2019104110-appb-100012
  8. 根据权利要求1所述的异吲哚啉酮类衍生物的制备方法，其特征在于，包括以下步骤：The method for preparing isoindolinone derivatives according to claim 1, characterized in that it comprises the following steps:
    Figure PCTCN2019104110-appb-100013
    Figure PCTCN2019104110-appb-100013
    由化合物VIII与叔丁基二甲基氯硅烷TBSCl反应制备化合物X；再由化合物X与IX经取代反应制备化合物XI；最后由化合物XI脱去羟基保护基后成盐制备化合物I .A。 Then replaced by compounds IX and X was prepared by reaction of compound XI;; from the compound VIII with t-butyl dimethyl chlorosilane compound X TBSCI prepared from compound XI end of the reaction after removal of hydroxy protecting groups to the preparation of salts of Compound I A..
  9. 一种药物组合物，其特征在于，包含权利要求1的通式I化合物或其药学上可接受的盐及药学上可接受的载体。A pharmaceutical composition, characterized by comprising the compound of general formula I according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
  10. 权利要求1所述异吲哚啉酮类衍生物或其药学上可接受的盐在制备ERK激酶抑制剂药物中的用途。Use of the isoindolinone derivatives or pharmaceutically acceptable salts thereof according to claim 1 in the preparation of ERK kinase inhibitor drugs.
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