A preparation method and intermediate of azaquinolinone derivatives

Abstract

The present application relates to the technical field of organic synthesis, and specifically to a method for preparing an azaquinolinone derivative, an intermediate M5 for preparing an azaquinolinone derivative, and a preparation method thereof. The preparation method of the intermediate M5 comprises: an intermediate M4 reacts with a sulfonylating agent or a halogenating agent to obtain the intermediate M5; the preparation method of the azaquinolinone derivative comprises: step a) intermediate M5 reacts with intermediate M7 to obtain compound 8; the preparation method of the azaquinolinone derivative is a process route with high yield, environmental friendliness, and impurity content within a limit range. The route can realize industrial production and amplification, is simple to operate, and the prepared azaquinolinone derivative has high purity.

Description

Technical Field

[0001] The present application relates to the technical field of organic synthesis, and in particular to a method for preparing an azaquinolinone derivative, an intermediate M5 for preparing an azaquinolinone derivative and a method for preparing the same. Background Art

[0002] PARP1 and PARP2 (PARP1 / 2) bind to DNA damage sites and catalyze the synthesis of poly ADP ribose chains on protein substrates, recruiting other DNA repair proteins to the damage sites to jointly repair DNA damage, while triggering the release of PARP1 / 2 from DNA. PARP inhibitors "capture" PARP1 / 2 to DNA by binding to the catalytic site of PARP1 / 2, causing PARP1 / 2 to be unable to fall off from the DNA damage site, thereby inducing DNA replication fork stagnation and DNA replication failure. At this time, cells usually repair this error through homologous repeat repair (HRR). In the HRR process, proteins such as BRCA1 / 2 play an important role. In cells with BRCA mutations, HRR function is abnormal, and the presence of PARP inhibitors will prevent DNA damage repair and lead to cell death.

[0003] Olaparib is the world's first approved PARP inhibitor developed by AstraZeneca and has been approved by the FDA for the treatment of multiple cancers. Since then, rucaparib, niraparib and talazoparib have also been approved for marketing; at the same time, there are several inhibitors in the clinical stage. Although PARP inhibitors have shown excellent clinical efficacy in patients with BRCA deficiency, these compounds have shown significant hematologic toxicity, including anemia, neutropenia and thrombocytopenia, whether used as a single agent or in combination therapy. Hematologic toxicity limits the use of first-generation PARP inhibitors, and clinically requires dose reduction, suspension of administration or termination of medication.

[0004] Recent studies have shown that PARP2 inhibition is closely related to blood toxicity, that PARP2, rather than PARP1, is essential for the survival of mouse hematopoietic stem / progenitor cells (HSPCs) and is used to maintain hematopoietic homeostasis, and that the synthetic lethality of BRCA mutations is driven only by PARP1 and does not require PARP2 to be "captured" to DNA. Since PARP1 and PARP2 are highly homologous, most current PARP inhibitors lack selectivity for PARP1. Therefore, the development of highly selective PARP1 inhibitors is beneficial for reducing blood toxicity and improving the therapeutic index.

[0005] Compared with other PARP1 / 2 inhibitors, highly selective PARP1 inhibitors are expected to improve efficacy and reduce toxicity. Therefore, there is an unmet medical need for effective and safe PARP inhibitors. Currently, there are no highly selective PARP1 inhibitors on the market. The selective PARP1 inhibitor AZD5305 has entered clinical phase I / II, and the PARP1 inhibitor AZD9574, which is more selective than AZD5305, has also entered clinical phase I / II.

[0006] There are still uncertainties in the effectiveness, safety, etc. of the compounds and experimental drugs disclosed in the prior art. Therefore, it is still necessary to find a method for preparing compounds used as PARP1 inhibitors that have excellent performance in terms of effectiveness, safety, selectivity, etc. Summary of the invention

[0007] The applicant has developed an azaquinolinone compound, which is recorded in the patent PCT / CN2023 / 097403 filed on May 31, 2023, specifically its Example 6 compound 5-(4-((8-fluoro-2-methyl-3-oxo-3,4-dihydroquinoxaline-6-yl)methyl)piperazine-1-yl)-N,6-dimethylpyridineamide (6) (i.e., compound 8 of the present application), which has excellent performance in terms of effectiveness, safety and selectivity, and is a compound used as a PARP1 inhibitor, and the structural formula is as follows:

[0008]

[0009] Based on this, the applicant has developed an intermediate M5 for preparing compound 8 and its preparation method, as well as a process route with high yield, environmental friendliness, and impurity content within the limit. This route can realize the industrial production scale-up of compound 8, is easy to operate, and the prepared compound 8 has high purity.

[0010] The present application provides an intermediate M5;

[0011]

[0012] In the intermediate M5, X is selected from OTs, OMs, OTf, Cl, Br, I, p-nitrobenzenesulfonyloxy, 2,5-dichlorobenzenesulfonyloxy or 4-chlorobenzenesulfonyloxy;

[0013] Preferably, X is selected from OTs, OMs, OTf, Cl, Br, I;

[0014] X is preferably Br or X is preferably OTs.

[0015] In some embodiments, the intermediate M5 is an intermediate for preparing azaquinolinone compound 8.

[0016] The present application provides a method for preparing an intermediate M5, which comprises:

[0017]

[0018] wherein X is selected from OTs, OMs, OTf, Cl, Br, I, p-nitrobenzenesulfonyloxy, 2,5-dichlorobenzenesulfonyloxy and 4-chlorobenzenesulfonyloxy;

[0019] Preferably, X is selected from OTs, OMs, OTf, Cl, Br, I; X is preferably Br or X is preferably OTs;

[0020] Preferably, the sulfonylating agent is selected from methanesulfonyl chloride, phenylmethanesulfonyl chloride, p-toluenesulfonyl chloride, trifluoromethanesulfonyl chloride, trifluoromethanesulfonic anhydride, p-nitrobenzenesulfonyl chloride, 2,5-dichlorobenzenesulfonyl chloride and 4-chlorobenzenesulfonyl chloride, preferably p-toluenesulfonyl chloride;

[0021] Preferably, the halogenating agent is selected from halogen, hydrogen halide, carbon tetrahalide, sulfoxide halide, phosphorus halide, triphenylphosphine, organic phosphine halide, N-haloamide and halogenated salt, preferably selected from carbon tetrabromide, chlorine, thionyl chloride, oxalyl chloride, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, hydrochloric acid, hydrogen chloride, triphenylphosphine, triphenylphosphine chloride, bromine, phosphorus tribromide, N-bromosuccinimide, triphenylphosphine bromide, N-iodosuccinimide and elemental iodine, preferably carbon tetrabromide and / or triphenylphosphine;

[0022] Preferably, the molar ratio of the intermediate M4 to the sulfonylating agent is 1:1-5, for example: 1:1, 1:1.5, 1:5; preferably 1:1.5;

[0023] Preferably, the molar ratio of the intermediate M4 to the halogenating agent is 1:1-10, for example: 1:1, 1:3, 1:6, 1:10; preferably 1:3 or 1:6;

[0024] Preferably, the step d) specifically comprises: under alkaline conditions, the intermediate M4 reacts with the sulfonylating agent to obtain the intermediate M5;

[0025] The base is preferably an inorganic base or an organic base; the inorganic base is preferably an alkali metal salt; the alkali metal salt is preferably one or more of alkali metal hydroxides, alkali metal carbonates, alkali metal bicarbonates and alkali metal phosphates; the organic base is preferably one or more of alkali metal salts of alcohols, lithium amide compounds, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), triethylamine, 4-dimethylaminopyridine and 1,8-diazabicyclo[5.4.0]undec-7-ene; the alkali metal salts of alcohols are preferably one or more of sodium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide; the lithium amide compound is preferably lithium diisopropylamide (LDA) or lithium hexamethyldisilazide (LiHMDS);

[0026] The base is more preferably an organic base, and further preferably N,N-diisopropylethylamine (DIPEA) and / or 4-dimethylaminopyridine (DMAP);

[0027] Preferably, in the step d), the molar ratio of the intermediate M4 to the base is 1:2;

[0028] Preferably, the base is N,N-diisopropylethylamine (DIPEA) and 4-dimethylaminopyridine (DMAP); the molar ratio of the intermediate M4 to the N,N-diisopropylethylamine (DIPEA) is 1:1-5, for example: 1:1, 1:2, 1:5; preferably 1:2; the molar ratio of the intermediate M4 to 4-dimethylaminopyridine (DMAP) is 1:0.01-0.2, for example: 1:0.01, 1:0.05, 1:0.2; preferably 1:0.05;

[0029] Preferably, the step d) specifically comprises: reacting the intermediate M4 with the sulfonylating agent or the halogenating agent in a solvent D to obtain the intermediate M5;

[0030] Preferably, the step d) specifically comprises: reacting the intermediate M4 with p-toluenesulfonyl chloride in a solvent D to obtain the intermediate M5;

[0031] The solvent D is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate;

[0032] The solvent D is more preferably dimethylformamide (DMF) or dichloromethane;

[0033] In some embodiments, in the step d), the intermediate M4 reacts with the sulfonylating agent, and the mass volume ratio of the intermediate M4 to the solvent D is 1 g:5 ml; the solvent D is preferably dimethylformamide (DMF);

[0034] In some embodiments, in the step d), the intermediate M4 reacts with the halogenating agent, and the mass volume ratio of the intermediate M4 to the solvent D is 1 g:29.4 ml; the solvent D is preferably dichloromethane;

[0035] Preferably, the preparation method of step d) specifically comprises: under alkaline conditions, the intermediate M4 and the sulfonylating agent react in solvent D to obtain the intermediate M5;

[0036] Preferably, the preparation method in step d) specifically comprises: reacting the intermediate M4 with the halogenating agent in a solvent D to obtain the intermediate M5;

[0037] Preferably, the reaction temperature of step d) is 0°C-100°C; the intermediate M4 reacts with the sulfonylating agent, and the reaction temperature is preferably room temperature; the intermediate M4 reacts with the halogenating agent, and the reaction temperature is preferably 0°C;

[0038] In some embodiments, the preparation method of step d) specifically comprises: adding a base and the intermediate M4 into a solvent D, cooling, adding the toluenesulfonyl chloride, and heating to react to obtain the intermediate M5;

[0039] In some embodiments, the preparation method of step d) specifically comprises: adding a base and the intermediate M4 into a solvent D, cooling to 0° C., adding to the toluenesulfonyl chloride, heating to room temperature for reaction, and obtaining the intermediate M5;

[0040] In some specific embodiments, the step d) specifically comprises: under nitrogen protection, adding intermediate M4 (1.0 g, 4.8 mmol), DIPEA (1.24 g, 9.6 mmol), and DMAP (30 mg, 0.24 mmol) to DMF (5 mL), stirring and cooling to 0°C; dissolving p-toluenesulfonyl chloride (1.37 g, 7.2 mmol) in DMF (5 mL), dripping into the reaction solution, and warming to room temperature for reaction;

[0041] In some specific embodiments, the step d) is specifically as follows: under nitrogen protection, 4.8 mmol intermediate M4, 9.6 mmol DIPEA, and 0.24 mmol DMAP are added to 5 mL DMF, stirred and cooled to 0°C to obtain a reaction solution; 7.2 mmol p-toluenesulfonyl chloride is dissolved in 5 mL DMF and added dropwise to the reaction solution; after the addition is completed, the temperature is raised to room temperature for reaction; after the reaction is complete, the mixture is diluted with water, extracted three times with ethyl acetate, the organic phases are combined, and washed three times with 10% brine; the liquids are separated, the organic phase is dried, filtered, concentrated under reduced pressure, and subjected to column chromatography to obtain the intermediate M5.

[0042] In some embodiments, the intermediate M4 and triphenylphosphine are dissolved in solvent D, carbon tetrabromide is added, and the reaction is carried out at 0°C.

[0043] In some specific embodiments, the step d) specifically comprises: at 0°C, intermediate M4 (1.7 g, 8.2 mmol) and triphenylphosphine (6.4 g, 24.6 mmol) are dissolved in dichloromethane (50 mL), stirred for 0.5 h, and heated to 40°C. 2 Add carbon tetrabromide (8.2 g, 24.6 mmol) under protection, and react at 0°C overnight;

[0044] In some embodiments, the step d) further comprises: washing, drying, filtering, concentrating under reduced pressure, and column chromatography after the reaction is complete;

[0045] In some specific embodiments, the step d) further comprises: monitoring by TLC until the reaction is complete, diluting with water, extracting with ethyl acetate three times, combining the organic phases, washing with 10% saline three times; separating the liquids, drying the organic phase, filtering, concentrating under reduced pressure, and column chromatography to obtain intermediate M5;

[0046] In some specific embodiments, the step d) further comprises: monitoring by TLC until the reaction is complete, concentrating the reaction system to obtain a crude compound, and subjecting the obtained crude product to silica gel column chromatography to obtain intermediate M5.

[0047] In some specific embodiments, the step d) is specifically as follows: at 0°C, 8.2 mmol of intermediate M4 and 24.6 mmol of triphenylphosphine are dissolved in 50 mL of dichloromethane, stirred for 0.5 h, 24.6 mmol of carbon tetrabromide is added under nitrogen protection, and the reaction is carried out at 0°C overnight. After the reaction is complete, the reaction system is concentrated to obtain a crude compound, and the obtained crude product is subjected to silica gel column chromatography to obtain intermediate M5.

[0048] The preparation method of intermediate M5 of the present application also includes:

[0049] Step e) reacting the intermediate M3 with (tributyltin)methanol to obtain the intermediate M4;

[0050]

[0051] Preferably, the molar ratio of the intermediate M3 to the (tributyltin)methanol is 1:1-3, preferably 1:1.2;

[0052] Preferably, the step e) specifically comprises: reacting the intermediate M3 and the (tributyltin)methanol in a solvent E to obtain the intermediate M4;

[0053] The solvent E is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate;

[0054] The solvent E is more preferably 1,4-dioxane;

[0055] Preferably, the mass volume ratio of the intermediate M3 and the solvent E is 1g:5-20ml, for example, 1g:5ml, 1g:10ml, 1g:20ml; preferably 1g:10ml;

[0056] In some embodiments, in the step e), the mass volume ratio of the intermediate M3 and the solvent E is 1 g:10 ml.

[0057] Preferably, the preparation method in step e) specifically comprises: under the condition of catalyst E, the intermediate M3 and the (tributyltin)methanol react to obtain the intermediate M4;

[0058] The catalyst E is preferably a metal catalyst, and the metal catalyst is preferably any one of a palladium catalyst, a cobalt catalyst and a nickel catalyst; for example, the palladium catalyst is selected from tris(dibenzylideneacetone)dipalladium, chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II), palladium acetate, palladium propionate, palladium oxide, palladium nitrate, palladium sulfate, palladium trifluoroacetate, palladium acetylacetonate, palladium pivalate, palladium dichloride, palladium dibromide, Palladium diiodide, tetraamminepalladium dichloride, tetraamminepalladium acetate, palladium hydroxide, palladium on carbon, tetrakis(triphenylphosphine)palladium, (1,1′-bis(diphenylphosphinoferrocene)palladium dichloride, bis(triphenylphosphine)palladium dichloride, bis(tricyclohexylphosphine)palladium dichloride, bis(tri-tert-butylphosphine)palladium, (1,3-bis(diphenylphosphino)propane)palladium dichloride, (1,5-cyclooctadiene)palladium dichloride, (2,2′-bipyridine)palladium dichloride, 1,2-bis(diphenylphosphino)ethanepalladium dichloride, 1,4-bis(diphenylphosphino)butane Palladium dichloride, bis(triphenylphosphine)palladium acetate, ethylenediamine palladium dichloride, bis(cyanobenzene)palladium dichloride, 1,2-bis(phenylsulfinyl)ethane diacetate palladium, (1,10-phenanthroline)palladium dichloride, bis(tri-o-tolylphosphine)palladium dichloride, hexafluoroacetylacetonate palladium, bis[1,2-bis(diphenylphosphino)ethane]palladium, bis[tri(2-tolyl)phosphine]palladium, bis(triethylphosphine)palladium dichloride, bis(acetonitrile)palladium dichloride, bis(dibenzylideneacetone)palladium, bis(methyldiphenylphosphine)palladium dichloride, tetra( One or more of acetonitrile) palladium tetrafluoroborate, (bicyclo[2.2.1]hepta-2,5-diene) dichloropalladium, 1,1′-bis(di-tert-butylphosphino)ferrocene dichloropalladium, 1,1′-bis(dicyclohexylphosphino)ferrocene dichloropalladium, 1,1′-bis(diisopropylphosphino)ferrocene dichloropalladium, (N,N,N′,N′-tetramethylethylenediamine) dichloropalladium, bis[dicyclohexyl(4-dimethylaminophenyl)phosphine] dichloropalladium, bis[di-tert-butyl(4-trifluoromethylphenyl)phosphine] dichloropalladium;

[0059] The nickel catalyst is selected from one or more of bis(triphenylphosphine)nickel chloride, nickel chloride, dichlorobis(tributylphosphine)nickel, dichlorobis(trimethylphosphine)nickel, bis(tributylphosphine)nickel dibromide, bis(tricyclohexylphosphine)nickel dichloride, bis(triphenylphosphine)nickel dibromide, bis(triphenylphosphine)nickel dicarbonyl, bis(triphenylphosphine)nickel chloride, [1,3-bis(diphenylphosphine)propane]nickel dichloride, [1,2-bis(diphenylphosphine)ethane]nickel dichloride, [1,1′-bis(diphenylphosphine)ferrocene]nickel dichloride, bis(isopropylcyclopentadiene)nickel, and tetrakis(triphenylphosphine)nickel;

[0060] The cobalt catalyst is selected from one or more of tris(triphenylphosphine)cobalt chloride, bis(triphenylphosphine)cobalt chloride, diiodo(bis(diphenylphosphine)ethane)cobalt, bis(ethylcyclopentadiene)cobalt hexafluorophosphate, [1,2′-bis(diphenylphosphine)ethane]cobalt dichloride, [1,1′-bis(diphenylphosphine)ferrocene]cobalt dichloride, and bis(pentamethylcyclopentadienyl)cobalt hexafluorophosphate;

[0061] The catalyst E is more preferably chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (Xphos Pd G2);

[0062] Preferably, the molar ratio of the intermediate M3 to the catalyst E is 5-30:1, for example: 5:1, 20:1, 30:1; preferably 20:1.

[0063] Preferably, the preparation method in step e) specifically comprises: under the condition of catalyst E, the intermediate M3 and the (tributyltin)methanol react in solvent E to obtain the intermediate M4;

[0064] In some embodiments, in the step e), the molar ratio of the intermediate M3 to the catalyst E is 1:50.

[0065] Preferably, the reaction temperature of step e) is 60°C-120°C, preferably 90°C-100°C, such as 90°C, 95°C, 100°C;

[0066] Preferably, the reaction time of step e) is 12h-16h.

[0067] In some specific embodiments, the step e) specifically comprises: dissolving the intermediate M3 (61.7 g, 0.24 mol) and (tributyltin)methanol (92.5 g, 0.29 mol) in 1,4-dioxane (617 mL), 2 Xphos PdG2 (9.4 g, 12 mmol) was added under protection, and the temperature was raised to 95±5°C and reacted overnight.

[0068] In some embodiments, the step e) further comprises: after the reaction is complete, the mixture is concentrated to dryness, methyl tert-butyl ether is added to the residue, and the mixture is refluxed, cooled, filtered, and dried under reduced pressure.

[0069] In some specific embodiments, the step e) further comprises: monitoring by LC until the reaction is complete, concentrating the reaction system to dryness, adding 1553 ml of methyl tert-butyl ether to the residue, refluxing and stirring for 1 hour, cooling to room temperature and filtering, and drying under reduced pressure to obtain intermediate M4.

[0070] In some specific embodiments, the step e) is specifically as follows: 0.24 mol of intermediate M3 and 0.29 mol of (tributyltin) methanol are dissolved in 617 mL of 1,4-dioxane, N 2 12 mmol Xphos Pd G2 was added under protection, and the temperature was raised to 95±5°C for overnight reaction. After the reaction was complete, the reaction system was concentrated to dryness, and 1553 ml of methyl tert-butyl ether was added to the residue, refluxed with stirring for 1 h, cooled to room temperature, filtered, and dried under reduced pressure to obtain intermediate M4.

[0071] The preparation method of intermediate M5 of the present application also includes:

[0072] Dehydrogenation reaction of the intermediate M2 to obtain the intermediate M3;

[0073]

[0074] Preferably, the step f) specifically comprises: dehydrogenating the intermediate M2 in a solvent F to obtain the intermediate M3;

[0075] The solvent F is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate;

[0076] The solvent F is more preferably 1,4-dioxane;

[0077] Preferably, the mass to volume ratio of the intermediate M2 and the solvent F is 1g:5-30ml, for example: 1g:5ml, 1g:20ml, 1g:30ml; preferably 1g:20ml.

[0078] In some embodiments, in step f), the mass volume ratio of the intermediate M2 and the solvent F is 1 g: 20 ml;

[0079] Preferably, the preparation method in step f) specifically comprises: under oxidizing conditions, dehydrogenating the intermediate M2 to obtain the intermediate M3;

[0080] The oxidant is preferably 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ);

[0081] In some embodiments, the molar ratio of the intermediate M2 to the oxidant is 1:1-4, for example: 1:1, 1:1.175, 1:4; preferably 1:1.175;

[0082] In some embodiments, the preparation method of step f) specifically comprises: dissolving the intermediate M2 in solvent F1 to obtain a mixed solution F1; dissolving an oxidant in solvent F2 to obtain a mixed solution F2; and adding the mixed solution F2 to the mixed solution F1.

[0083] In some embodiments, the preparation method of step f) specifically comprises: dissolving the intermediate M2 in solvent F1, cooling the temperature to 0°C to obtain a mixed solution F1; dissolving an oxidant in solvent F2 to obtain a mixed solution F2; dripping the mixed solution F2 into the mixed solution F1, and reacting at room temperature overnight;

[0084] In some embodiments, the solvent F1 and the solvent F2 are each independently selected from one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate;

[0085] In some embodiments, the intermediate M2 and the solvent F1 have a mass volume ratio of 1 g:10 ml;

[0086] In some embodiments, the oxidant and the solvent F2 have a mass volume ratio of 1 g:7.8 ml;

[0087] The oxidant is preferably 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) or manganese dioxide, preferably 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ);

[0088] Preferably, the reaction temperature of step f) is 0-100°C, preferably room temperature;

[0089] Preferably, the reaction time of step f) is 12h-16h.

[0090] In some specific embodiments, the step f) specifically includes: the intermediate M2 (102.0 g, 0.40 mol) is dissolved in 1,4-dioxane (1020 mL), the temperature is lowered to 0°C after nitrogen replacement, DDQ (131.4 g, 0.47 mol) is dissolved in 1,4-dioxane (1020 mL), and the mixture is slowly dripped into the reaction system, and the reaction is carried out at room temperature overnight.

[0091] In some embodiments, the step f) further comprises: hot pulping, filtering, and reduced pressure drying processes.

[0092] In some specific embodiments, the step f) further comprises: monitoring the reaction to be complete by TLC, cooling to 5±5°C, slowly adding 3L of saturated sodium bicarbonate aqueous solution, stirring at this temperature for 1h, and filtering. Add 2L of saturated sodium bicarbonate aqueous solution to the solid, heat to 50±5°C, heat for 1h, cool to room temperature, and filter. Repeat the above hot pulping process (add 2L of saturated sodium bicarbonate aqueous solution to the solid, heat to 50±5°C, heat for 1h, cool to room temperature, and filter) 5 times, filter, and dry the solid under reduced pressure to obtain intermediate M3.

[0093] In some specific embodiments, the preparation method of step f) is specifically as follows: 0.40 mol of intermediate M2 is dissolved in 1020 mL of 1,4-dioxane, the temperature is lowered to 0°C after nitrogen replacement, 0.47 mol of DDQ is dissolved in 1020 mL of 1,4-dioxane, and the mixture is slowly dripped into the reaction system; after the dripping, the mixture is reacted at room temperature overnight, and after the reaction is complete, the temperature is lowered to 5±5°C, 3 L of saturated sodium bicarbonate aqueous solution is dripped, the mixture is stirred for 1 hour at the temperature, and then filtered to obtain a solid, and the obtained solid is subjected to a hot pulping process, the hot pulping process is repeated 5 times, filtered, and the solid is dried under reduced pressure to obtain an intermediate M3.

[0094] The preparation method of intermediate M5 of the present application also includes:

[0095] Step g) reacting the intermediate M1 to obtain the intermediate M2;

[0096]

[0097] Preferably, the step g) specifically comprises: reacting the intermediate M1 in a solvent G to obtain the intermediate M2;

[0098] The solvent G is preferably a mixed solvent of glacial acetic acid and water; the volume ratio of glacial acetic acid to water is preferably 5-12:1, for example: 5:1, 9:1, 12:1; preferably 9:1;

[0099] The mass volume ratio of the intermediate M1 and the solvent G is 1g:5-20ml, for example: 1g:5ml, 1g:10ml, 1g:20ml; preferably 1g:10ml;

[0100] Preferably, the step g) specifically comprises: under the condition of catalyst G, the intermediate M1 reacts to obtain the intermediate M2;

[0101] The catalyst G is preferably zinc powder, palladium carbon, iron powder, tin dichloride, ferrous sulfate, sodium dithionite, and more preferably zinc powder;

[0102] The molar ratio of the solvent G to the catalyst G is 1:3-10, preferably 1:5;

[0103] Preferably, the step g) specifically comprises: under the condition of catalyst G, the intermediate M1 reacts in solvent G to obtain the intermediate M2;

[0104] Preferably, the reaction temperature of step g) is 50-100°C, preferably 90°C to 100°C, such as 90°C, 95°C, 100°C;

[0105] Preferably, the reaction time of step g) is 12h-16h.

[0106] In some specific embodiments of the present invention, the step g) specifically includes dissolving the intermediate M1 (138.0 g, 0.43 mol) in glacial acetic acid / water (1242 mL / 138 mL), cooling the reaction system to 0°C, adding zinc powder (140.5 g, 2.15 mol) in batches, replacing nitrogen after the addition, and raising the reaction system to 95±5°C for overnight reaction.

[0107] In some embodiments of the present invention, the step g) further comprises: extraction, washing, and concentration processes.

[0108] In some specific embodiments of the present invention, the step g) further comprises: using LC-MS to monitor the reaction until completion, cooling the reaction system to room temperature and then diluting with water, extracting three times with ethyl acetate, combining the organic phases, washing the organic phases with saturated sodium bicarbonate aqueous solution until the aqueous phase is weakly alkaline, and concentrating the organic phase to obtain the intermediate M2.

[0109] In some specific embodiments of the present invention, the step g) is specifically 0.43 mol of intermediate M1 (138.0 g, 0.43 mol) is dissolved in glacial acetic acid / water (1242 mL / 138 mL), the reaction system is cooled to 0°C, 2.15 mol of zinc powder is added, nitrogen is replaced after the addition, the reaction system is heated to 95±5°C and reacted overnight, the reaction is completed, the reaction system is cooled to room temperature and diluted with water, extracted three times with ethyl acetate, the organic phases are combined, and the organic phases are washed with saturated sodium bicarbonate aqueous solution until the aqueous phase is weakly alkaline, and the organic phase is concentrated to obtain intermediate M2.

[0110] The preparation method of intermediate M5 of the present application also includes:

[0111] Step h) reacting the intermediate SM1 with alanine methyl ester hydrochloride to obtain the intermediate M1;

[0112]

[0113] Preferably, the molar ratio of the compound SM1 to the alanine methyl ester hydrochloride is 1:1-3, for example: 1:1, 1:1.1, 1:1.5, 1:3; preferably 1:1-1.5; more preferably 1:1.1.

[0114] Preferably, the step h) specifically comprises: under alkaline conditions, reacting the compound SM1 with the alanine methyl ester hydrochloride to obtain the intermediate M1;

[0115] The base is preferably an organic base; the organic base is preferably N,N-diisopropylethylamine (DIPEA), sodium tert-butoxide, tert-butylmagnesium chloride, triethylamine, pyridine or 4-dimethylaminopyridine (DMAP);

[0116] The base is more preferably N,N-diisopropylethylamine (DIPEA);

[0117] In some embodiments, in step g), the molar ratio of the compound SM1 to the base is 1:3;

[0118] Preferably, the step h) specifically comprises: reacting the compound SM1 and the alanine methyl ester hydrochloride in a solvent H to obtain the intermediate M1;

[0119] The solvent H is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate;

[0120] The solvent H is more preferably 1,4-dioxane;

[0121] In some embodiments, in the step h), the mass volume ratio of the compound SM1 to the solvent H is 1 g:1.4 ml.

[0122] Preferably, the preparation method of step h) specifically comprises: under alkaline conditions, the compound SM1 and the alanine methyl ester hydrochloride;

[0123] Preferably, the reaction temperature of step h) is 20°C-100°C, preferably 55°C-65°C, for example: 55°C, 60°C, 65°C;

[0124] Preferably, the reaction time of step h) is 2-10 h, for example: 2 h, 3 h, 10 h; preferably 3 h;

[0125] In some specific embodiments, the step h) specifically comprises: dissolving the compound SM1 (100.0 g, 0.42 mol) and the alanine methyl ester hydrochloride (64.5 g, 0.46 mol) in 1,4-dioxane (140 mL), adding DIPEA (163 g, 1.26 mol), and heating the reaction system to 60±5° C. for 3 h after the addition;

[0126] In some embodiments, the step h) further comprises: extraction, washing, and drying after the reaction is complete;

[0127] In some specific embodiments, the step h) further comprises: after the reaction is complete, concentrating the reaction system, diluting with water, extracting with ethyl acetate three times, combining the organic phases, washing with 10% sodium chloride aqueous solution three times (500 ml*3), and drying the organic phase and concentrating to obtain the intermediate M1.

[0128] In some specific embodiments, the step h) is specifically as follows: 0.42 mol of compound SM1 and 0.46 mol of alanine methyl ester hydrochloride are dissolved in 140 mL of 1,4-dioxane, 1.26 mol of DIPEA is added, and the reaction system is heated to 60±5° C. for 3 hours after the addition is completed. After the reaction is complete, the reaction system is concentrated, diluted with water, extracted three times with ethyl acetate, the organic phases are combined, washed three times with 10% sodium chloride aqueous solution, and the organic phase is dried and concentrated to obtain the intermediate M1.

[0129] The present application provides a method for preparing an intermediate M7, which comprises:

[0130] Step b) reacting the intermediate M6 with tert-butyl piperazine-1-carboxylate to obtain the compound M7;

[0131]

[0132] Preferably, the molar ratio of the intermediate M6 to the piperazine-1-carboxylic acid tert-butyl ester is 1:1-5, preferably 1:2;

[0133] Preferably, the step b) specifically comprises: under alkaline conditions, the intermediate M6 and the piperazine-1-carboxylic acid tert-butyl ester react to obtain the compound M7;

[0134] The base is preferably an inorganic base or an organic base; the inorganic base is preferably an alkali metal salt; the alkali metal salt is preferably one or more of alkali metal hydroxides, alkali metal carbonates, alkali metal bicarbonates and alkali metal phosphates; the organic base is preferably one or more of alkali metal salts of alcohols, amide lithium compounds, amide lithium compounds, N,N-diisopropylethylamine, triethylamine, 4-dimethylaminopyridine and 1,8-diazabicyclo[5.4.0]undec-7-ene; the alkali metal salts of alcohols are preferably one or more of sodium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide; the amide lithium compound is preferably lithium diisopropylamide (LDA) or lithium hexamethyldisilazide (LiHMDS);

[0135] The base is more preferably an inorganic base; further preferably cesium carbonate;

[0136] In some embodiments, in step b), the molar ratio of the intermediate M6 to the base is 1:3;

[0137] Preferably, the step b) specifically comprises: reacting the intermediate M6 and the piperazine-1-carboxylic acid tert-butyl ester in solvent B to obtain the compound 8;

[0138] The solvent B is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate;

[0139] The solvent B is more preferably toluene;

[0140] In some embodiments, in the step b), the mass volume ratio of the intermediate M6 and the solvent B is 1 g: 20.4 ml;

[0141] Preferably, the preparation method of step b) specifically comprises: reacting the intermediate M6 with the piperazine-1-carboxylic acid tert-butyl ester under the condition of catalyst B to obtain the compound 8;

[0142] The catalyst B is preferably a metal catalyst, and the metal catalyst is preferably any one of a palladium catalyst, a cobalt catalyst and a nickel catalyst; for example, the palladium catalyst is selected from tris(dibenzylideneacetone)dipalladium, chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II), palladium acetate, palladium propionate, palladium oxide, palladium nitrate, palladium sulfate, palladium trifluoroacetate, palladium acetylacetonate, palladium pivalate, palladium dichloride, palladium dibromide, Palladium diiodide, tetraamminepalladium dichloride, tetraamminepalladium acetate, palladium hydroxide, palladium on carbon, tetrakis(triphenylphosphine)palladium, (1,1′-bis(diphenylphosphinoferrocene)palladium dichloride, bis(triphenylphosphine)palladium dichloride, bis(tricyclohexylphosphine)palladium dichloride, bis(tri-tert-butylphosphine)palladium, (1,3-bis(diphenylphosphino)propane)palladium dichloride, (1,5-cyclooctadiene)palladium dichloride, (2,2′-bipyridine)palladium dichloride, 1,2-bis(diphenylphosphino)ethanepalladium dichloride, 1,4-bis(diphenylphosphino)butane Palladium dichloride, bis(triphenylphosphine)palladium acetate, ethylenediamine palladium dichloride, bis(cyanobenzene)palladium dichloride, 1,2-bis(phenylsulfinyl)ethane diacetate palladium, (1,10-phenanthroline)palladium dichloride, bis(tri-o-tolylphosphine)palladium dichloride, hexafluoroacetylacetonate palladium, bis[1,2-bis(diphenylphosphino)ethane]palladium, bis[tri(2-tolyl)phosphine]palladium, bis(triethylphosphine)palladium dichloride, bis(acetonitrile)palladium dichloride, bis(dibenzylideneacetone)palladium, bis(methyldiphenylphosphine)palladium dichloride, tetra( One or more of acetonitrile) palladium tetrafluoroborate, (bicyclo[2.2.1]hepta-2,5-diene) dichloropalladium, 1,1′-bis(di-tert-butylphosphino)ferrocene dichloropalladium, 1,1′-bis(dicyclohexylphosphino)ferrocene dichloropalladium, 1,1′-bis(diisopropylphosphino)ferrocene dichloropalladium, (N,N,N′,N′-tetramethylethylenediamine) dichloropalladium, bis[dicyclohexyl(4-dimethylaminophenyl)phosphine] dichloropalladium, bis[di-tert-butyl(4-trifluoromethylphenyl)phosphine] dichloropalladium;

[0143] The nickel catalyst is selected from one or more of bis(triphenylphosphine)nickel chloride, nickel chloride, dichlorobis(tributylphosphine)nickel, dichlorobis(trimethylphosphine)nickel, bis(tributylphosphine)nickel dibromide, bis(tricyclohexylphosphine)nickel dichloride, bis(triphenylphosphine)nickel dibromide, bis(triphenylphosphine)nickel dicarbonyl, bis(triphenylphosphine)nickel chloride, [1,3-bis(diphenylphosphine)propane]nickel dichloride, [1,2-bis(diphenylphosphine)ethane]nickel dichloride, [1,1′-bis(diphenylphosphine)ferrocene]nickel dichloride, bis(isopropylcyclopentadiene)nickel, and tetrakis(triphenylphosphine)nickel;

[0144] The cobalt catalyst is selected from one or more of tris(triphenylphosphine)cobalt chloride, bis(triphenylphosphine)cobalt chloride, diiodo(bis(diphenylphosphine)ethane)cobalt, bis(ethylcyclopentadiene)cobalt hexafluorophosphate, [1,2′-bis(diphenylphosphine)ethane]cobalt dichloride, [1,1′-bis(diphenylphosphine)ferrocene]cobalt dichloride, and bis(pentamethylcyclopentadienyl)cobalt hexafluorophosphate;

[0145] The catalyst B is more preferably tris(dibenzylideneacetone)dipalladium.

[0146] More preferably, the palladium catalyst, nickel catalyst or cobalt catalyst is used in combination with a ligand, and the ligand is preferably 1,1'-binaphthyl-2,2'-bisdiphenylphosphine, triphenylphosphine, triethylphosphine, tributylphosphine, tri-tert-butylphosphine, tricyclohexylphosphine, triphenoxyphosphine, tri(o-methylphenyl)phosphine, tri(p-methoxyphenyl)phosphine, methyldiphenylphosphine, dicyclohexylphenylphosphine, di-tert-butylphenylphosphine, dicyclohexyl(4-dimethylaminophenyl)phosphine, di-tert-butyl(4-trifluoromethylphenyl)phosphine, 1,2-bis(diphenylphosphino)ethane, 1 ,3-bis(diphenylphosphino)propane, 1,4-bis(diphenylphosphino)butane, 1,2-bis(phenylsulfinyl)ethane, 1,1′-bisdiphenylphosphinoferrocene, 1,1′-bis(di-tert-butylphosphino)ferrocene, 1,1′-bis(dicyclohexylphosphino)ferrocene, 1,1′-bis(diisopropylphosphino)ferrocene and 2,3-O-isopropylene-2,3-dihydroxy-1,4-bisdiphenylphosphinobutane; the ligand is more preferably R-(+)-1,1′-binaphthyl-2,2′-bisdiphenylphosphine;

[0147] Preferably, the preparation method of step b) specifically comprises: reacting the intermediate M6 and the piperazine-1-carboxylic acid tert-butyl ester in solvent B under the conditions of catalyst B and base to obtain the compound 8;

[0148] In some embodiments, in step b), the molar ratio of the intermediate M6 to the catalyst B is 19.9:1;

[0149] Preferably, the reaction temperature of step b) is 50°C-120°C, preferably 95°C.

[0150] In some specific embodiments, the step b) specifically comprises: adding intermediate M6 (13.7 g, 59.8 mmol), tert-butyl piperazine-1-carboxylate (22.3 g, 119.6 mmol), and cesium carbonate (58.5 g, 179.4 mmol) to 280 ml of toluene, replacing with nitrogen, adding tri(dibenzylideneacetone)dipalladium (2.7 g, 3 mmol), R-(+)-1,1'-binaphthyl-2,2'-bisdiphenylphosphine (3.7 g, 6 mmol), replacing with nitrogen again, and heating to 95°C with stirring.

[0151] In some embodiments, the step b) further comprises: washing, drying, filtering, and concentrating under reduced pressure after the reaction is complete.

[0152] In some specific embodiments, the step b) further comprises: TLC monitoring, after the reaction is complete, 1L of water is added to the reaction system, and extracted twice with ethyl acetate. The organic phases are combined and washed with water, 5% sodium dihydrogen phosphate aqueous solution, and 10% sodium chloride aqueous solution in sequence. The organic phase is separated, dried, filtered, and concentrated under reduced pressure, and the obtained solid is added to 700ml of ethyl acetate, 24g of hydrochloric acid is added, and stirred at room temperature. TLC monitoring, after the reaction is complete, 500ml of water is added, the liquid is separated, and the aqueous phase is washed twice with ethyl acetate (200ml*2). The liquid is separated, and the aqueous phase is adjusted to weak alkalinity with 10% potassium carbonate aqueous solution. Extract five times with dichloromethane (200ml*5), the organic phases are combined, washed with water, dried, filtered, and concentrated under reduced pressure to obtain intermediate M7.

[0153] The present application provides a method for preparing an intermediate M6, which comprises:

[0154] Step c) Compound SM2 reacts with methylamine hydrochloride to obtain the intermediate M6;

[0155]

[0156] Preferably, the molar ratio of the compound SM2 to the methylamine hydrochloride is 1:1-5, preferably 1:2;

[0157] Preferably, the step c) specifically comprises: under alkaline conditions, the compound SM2 reacts with the methylamine hydrochloride to obtain the intermediate M6;

[0158] The base is preferably an inorganic base or an organic base; the inorganic base is preferably an alkali metal salt; the alkali metal salt is preferably one or more of an alkali metal hydroxide, an alkali metal carbonate, an alkali metal bicarbonate and an alkali metal phosphate; the organic base is preferably one or more of an alkali metal salt of an alcohol, an amide lithium compound, an amide lithium compound, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), triethylamine, 4-dimethylaminopyridine and 1,8-diazabicyclo[5.4.0]undec-7-ene; the alkali metal salt of an alcohol is preferably one or more of sodium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide; the amide lithium compound is preferably lithium diisopropylamide (LDA) or lithium hexamethyldisilazide (LiHMDS); the base is more preferably an organic base, and further preferably N,N-diisopropylethylamine (DIPEA);

[0159] In some embodiments, in the step c), the molar ratio of the compound SM2 to the base is 1:3;

[0160] Preferably, the step c) specifically comprises: reacting the compound SM2 with the methylamine hydrochloride in a solvent C to obtain the intermediate M6;

[0161] The solvent C is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate;

[0162] The solvent C is more preferably dichloromethane;

[0163] In some embodiments, in the step c), the mass volume ratio of the compound SM2 and the solvent C is 1 g:10 ml.

[0164] Preferably, the step c) specifically comprises: reacting the compound SM2 with the methylamine hydrochloride under a condensing agent to obtain the intermediate M6;

[0165] The condensing agent is preferably one or more of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 1-propylphosphoric anhydride (T3P), 1-hydroxybenzotriazole (HOBT), benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) and diphenylphosphoryl azide (DPPA);

[0166] The condensing agent is more preferably 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU);

[0167] In some embodiments, in step c), the molar ratio of the compound SM2 to the condensing agent is 1:1.5;

[0168] Preferably, the preparation method of step c) specifically comprises: reacting the compound SM2 with the methylamine hydrochloride in a solvent C under the conditions of a condensing agent and a base to obtain the intermediate M6;

[0169] Preferably, the reaction temperature of step c) is 0°C-40°C, preferably room temperature;

[0170] More preferably, the preparation method of step c) specifically comprises: adding the condensing agent, the base and the compound SM2 into solvent C, cooling, adding the methylamine hydrochloride, and heating to react to obtain the intermediate M6;

[0171] In some embodiments, the preparation method of step c) specifically comprises: adding the condensing agent, the base and the compound SM2 into solvent C, cooling to 0°C, adding the methylamine hydrochloride, heating to room temperature for reaction, and obtaining the intermediate M6.

[0172] In some specific embodiments, compound SM2 (20 g, 92.6 mmol), DIPEA (35.9 g, 277.7 mmol), and HATU (52.8 g, 138.9 mmol) were dissolved in dichloromethane (200 mL), stirred and cooled to 0°C, and methylamine hydrochloride (12.5 g, 185 mmol) was added. After the addition, the temperature was raised to room temperature for reaction.

[0173] In some embodiments, the step c) further comprises: washing, drying, filtering, concentrating under reduced pressure, and column chromatography after the reaction is complete;

[0174] In some specific embodiments, the reaction is monitored by TLC until it is complete, 300 ml of water is added, the liquid is separated, and the organic phase is washed once with purified water, 10% potassium carbonate aqueous solution, and 10% saline. The organic phase is dried, filtered, concentrated under reduced pressure, and subjected to column chromatography to obtain intermediate M6.

[0175] The present application provides a method for preparing compound 8, which comprises:

[0176] Step a) the intermediate M5 and the intermediate M7 are reacted to obtain the compound 8;

[0177]

[0178] Preferably, the molar ratio of the intermediate M5 to the intermediate M7 is 0.80:1-1:1.3, preferably 1:0.85 or 1:1.2;

[0179] Preferably, the step a) specifically comprises: under alkaline conditions, the intermediate M5 and the intermediate M7 react to obtain the compound 8;

[0180] The base is preferably an inorganic base or an organic base; the inorganic base is preferably an alkali metal salt; the alkali metal salt is preferably one or more of alkali metal hydroxides, alkali metal carbonates, alkali metal bicarbonates and alkali metal phosphates; for example, one or more of sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide and sodium phosphate; the organic base is preferably one or more of alkali metal salts of alcohols, amide lithium compounds, amide lithium compounds, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), triethylamine, 4-dimethylaminopyridine and 1,8-diazabicyclo[5.4.0]undec-7-ene; the alkali metal salts of alcohols are preferably one or more of sodium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide; the amide lithium compound is preferably lithium diisopropylamide (LDA) or lithium hexamethyldisilazide (LiHMDS);

[0181] The base is more preferably an inorganic base; further preferably potassium carbonate;

[0182] The base is more preferably an organic base; further preferably N,N-diisopropylethylamine (DIPEA).

[0183] In the present application, the compound 8 is an azaquinolinone compound.

[0184] In some embodiments, in step a), the molar ratio of the intermediate M5 to the base is 1:1.31-1:2; for example: 1:1.31 or 1:2;

[0185] Preferably, the step a) specifically comprises: reacting the intermediate M5 and the intermediate M7 in solvent A to obtain the compound 8;

[0186] The solvent A is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate or isopropyl acetate;

[0187] The solvent A is more preferably dimethylformamide (DMF);

[0188] The solvent A is more preferably 1,4-dioxane.

[0189] In some embodiments, in the step a), the mass volume ratio of the intermediate M5 to the solvent A is 100 mg:1 ml.

[0190] Preferably, the preparation method of step a) specifically comprises: under alkaline conditions, the intermediate M5 and the intermediate M7 react in solvent A to obtain the compound 8;

[0191] Preferably, the reaction temperature of step a) is 40°C-110°C, preferably 45-80°C, more preferably 45°C or 80°C; preferably, the reaction temperature of step a) is 50°C-100°C, preferably 80°C.

[0192] In some specific embodiments, the step a) specifically comprises: adding intermediate M5 (200 mg, 0.55 mmol), intermediate M7 (110 mg, 0.47 mmol), and potassium carbonate (100 mg, 0.72 mmol) to 2 ml of DMF, heating to 80° C. and stirring; or

[0193] The step a) specifically comprises: adding 22.13 mmol of intermediate M5 and 26.56 mmol of intermediate M7 into 60 mL of 1,4-dioxane, stirring, adding 44.26 mmol of N,N-diisopropylethylamine, and heating to 45° C. to react to obtain the compound 8.

[0194] In some embodiments, the step a) further comprises: adding water after the reaction is complete, cooling, filtering, and column chromatography; or cooling the reaction system, keeping warm and stirring, filtering, beating, filtering, and drying the filter cake;

[0195] In some embodiments, the cooling in step a) is cooling to 25°C; or cooling to 15°C;

[0196] In some embodiments, the mass volume ratio of the intermediate M5 and the water added after the reaction is complete is 100 mg: 1 ml;

[0197] In some specific embodiments, the step a) further comprises: monitoring by TLC until the reaction is complete, adding 2 ml of water to the reaction system, cooling to 25° C., filtering, and performing column chromatography to obtain compound 8; or

[0198] The reaction was monitored by TLC until completion, the reaction system was cooled to 15°C, stirred for 2 h, filtered, the filter cake was slurried with 180 mL of water at 20°C for 2 h, filtered, and the filter cake was dried under reduced pressure at 50°C and vacuum degree ≤-0.080 MPa to constant weight to obtain compound 8.

[0199] In some specific embodiments, the step a) is specifically as follows: 0.55 mmol intermediate M5, 0.47 mmol intermediate M7, and 0.72 mmol potassium carbonate are added to 2 ml DMF, heated to 80° C. and stirred until the reaction is complete; 2 ml water is added to the reaction system, the temperature is lowered to 25° C., and filtered; and compound 8 is obtained by column chromatography; or

[0200] The step a) is specifically as follows: 22.13 mmol of intermediate M5 and 26.56 mmol of intermediate M7 are added to 60 mL of 1,4-dioxane, stirring is started, 44.26 mmol of N,N-diisopropylethylamine is added, the temperature is raised to 45° C. and stirred, and the reaction is completed. The reaction system is cooled to 15° C., stirred for 2 h at this temperature, filtered, the filter cake is slurried with 180 mL of water at 20° C. for 2 h, filtered, and the filter cake is dried under reduced pressure at a temperature of 50° C. and a vacuum degree of ≤-0.080 MPa to constant weight to obtain compound 8.

[0201] In some specific embodiments, the preparation method of compound 8 comprises:

[0202] The intermediate SM1 reacts with alanine methyl ester hydrochloride to obtain the intermediate M1;

[0203]

[0204] The intermediate M1 is reacted to obtain the intermediate M2;

[0205]

[0206] The intermediate M2 is reacted to obtain the intermediate M3;

[0207]

[0208] The intermediate M3 reacts with (tributyltin)methanol to obtain the intermediate M4;

[0209]

[0210] The intermediate M4 reacts with a sulfonylating agent or a halogenating agent to obtain an intermediate M5;

[0211]

[0212] Compound SM2 reacts with methylamine hydrochloride to obtain intermediate M6;

[0213]

[0214] The intermediate M6 reacts with piperazine-1-carboxylic acid tert-butyl ester to obtain compound M7;

[0215]

[0216] The intermediate M5 and the intermediate M7 react to obtain the compound 8;

[0217]

[0218] Beneficial effects of this application:

[0219] Provided are an intermediate M5 for preparing azaquinolinone derivatives and a preparation method thereof, and a preparation method of azaquinolinone derivatives. The preparation method of intermediate M5 comprises: intermediate M4 reacts with a sulfonylating agent or a halogenating agent to obtain intermediate M5; the preparation method of azaquinolinone derivatives comprises: step a) intermediate M5 reacts with intermediate M7 to obtain compound 8. Compound 8 has excellent performance in terms of effectiveness, safety, selectivity and other properties, and is a compound used as a PARP1 inhibitor.

[0220] The preparation method of the compound 8 provided in the present application can achieve production of kilograms or more, has a high reaction yield, a low impurity content, a high purity, is more environmentally friendly, and can be used for industrial scale-up production. DETAILED DESCRIPTION

[0221] In order to make the present application easier to understand, the present application will be described in detail below in conjunction with embodiments. These embodiments are only for illustration and are not limited to the scope of application of the present application.

[0222] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0223] Unless otherwise stated, the instruments, materials, supplies and reagents used in the following examples can be obtained through conventional commercial means.

[0224] The terms "include", "comprising" and "containing" and their equivalents should be understood as having an open, non-exclusive meaning, i.e., "including but not limited to", meaning that in addition to the listed elements, components and steps, other unspecified elements, components and steps may also be included.

[0225] Unless otherwise specified in the present application, the methods, materials, etc. used in the present application are all conventional and commonly used in the art, and can implement the technical solutions of the present application.

[0226] Unless otherwise specified, all reagents involved in this application can be obtained from commercial sources.

[0227] The structures of the compounds were determined by mass spectrometry (MS) or nuclear magnetic resonance (1HNMR).

[0228] H NMR ( 1HNMR) shifts (δ) are given in parts per million (ppm); H NMR spectra ( 1 HNMR) was measured using a Bruker AVANCE-400 NMR spectrometer, and the solvent was deuterated chloroform (CDCl 3 ) and deuterated dimethyl sulfoxide, the internal standard was tetramethylsilane (TMS), and the chemical shift was 10 -6 (ppm) is given as unit; C NMR 13 CNMR was measured using a Bruker AVANCE-400 NMR spectrometer. The solvent was deuterated dimethyl sulfoxide, the internal standard was tetramethylsilane (TMS), and the chemical shift was 10 -6 (ppm) is given as the unit.

[0229] Mass spectrometry (MS) was performed using a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Therm, model: Finnigan LCQ advantage MAX).

[0230] Thin layer silica gel uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel sheet.

[0231] Column chromatography generally uses Yantai Huanghai Silica Gel 200-300 mesh silica gel as the carrier.

[0232] Unless otherwise specified in this application, the reactions mentioned in this application are all 2 Protected or nitrogen atmosphere.

[0233] The term "N" in this application 2 "Protective" or "nitrogen atmosphere" means, for example, connecting the reaction bottle to a 1 L nitrogen balloon.

[0234] Unless otherwise specified in the present application, the solution mentioned in the reaction of the present application is an aqueous solution.

[0235] The term "room temperature" in this application refers to a temperature between 10°C and 25°C.

[0236] The term "reacting overnight" in this application refers to a reaction time of 12h-16h.

[0237] Unless otherwise specified in the present application, "%" in the embodiments refers to mass percentage.

[0238] The term "about" means within ±10% of the stated numerical value, preferably within ±5%, more preferably within ±2%.

[0239] The abbreviations used herein have the following meanings:

[0240] Table 1

[0241]

[0242] Example 1 Synthesis of Intermediate M1

[0243]

[0244] Steps:

[0245] Compound SM1 (100.0 g, 0.42 mol) and alanine methyl ester hydrochloride (64.5 g, 0.46 mol) were dissolved in 1,4-dioxane (140 mL), and DIPEA (163 g, 1.26 mol) was added. After the addition, the reaction system was heated to 60±5°C for 3 hours. After the reaction was complete, the reaction system was concentrated, diluted with water, extracted three times with ethyl acetate, the organic phases were combined, and washed three times with 10% sodium chloride aqueous solution (500 ml*3). The organic phase was dried and concentrated to obtain intermediate M1 (142.0 g, yield 105%).

[0246] 1 H NMR (400 MHz, DMSO-d 6 )δ8.02 (s, 1H), 7.75-7.71 (d, J=6, 1H), 4.58-4.52 (m, 1H), 3.65 (s, 3H), 1.45-1.43 (d, J=8, 3H).

[0247] Example 2 Synthesis of Intermediate M2

[0248]

[0249] Steps:

[0250] Intermediate M1 (138.0 g, 0.43 mol) was dissolved in glacial acetic acid / water (1242 mL / 138 mL), the reaction system was cooled to 0°C, zinc powder (140.5 g, 2.15 mol) was added in batches, nitrogen was replaced after addition, and the reaction system was heated to 95±5°C for overnight reaction. The reaction was monitored by LC-MS until complete, the reaction system was cooled to room temperature and diluted with water, extracted three times with ethyl acetate, the organic phases were combined, and the organic phases were washed with saturated sodium bicarbonate aqueous solution until the aqueous phase was weakly alkaline. The organic phase was concentrated to obtain intermediate M2 (103.8 g, yield 93.4%).

[0251] 1 H NMR (400 MHz, DMSO-d 6 ) δ7.42-7.39 (d, J=6, 1H), 7.24 (s, 1H), 2.39 (s, 3H).

[0252] Example 3 Synthesis of Intermediate M3

[0253]

[0254] Steps:

[0255] Intermediate M2 (102.0 g, 0.40 mol) was dissolved in 1,4-dioxane (1020 mL), and the temperature was lowered to 0 ° C after nitrogen replacement. DDQ (131.4 g, 0.47 mol) was dissolved in 1,4-dioxane (1020 mL) and slowly added to the reaction system. After the addition, the reaction was allowed to react overnight at room temperature. TLC monitored the reaction to be complete, cooled to 5±5 ° C, 3 L of saturated sodium bicarbonate aqueous solution was slowly added, and the mixture was stirred for 1 h before filtering to obtain a solid. The obtained solid was added with 2 L of saturated sodium bicarbonate aqueous solution, heated to 50±5 ° C for 1 h, cooled to room temperature and filtered. Repeat the above hot pulping process (add 2 L of saturated sodium bicarbonate aqueous solution to the solid, heat to 50±5 ° C for 1 h, and filter after cooling to room temperature) 5 times. Filter, and dry the solid under reduced pressure to obtain intermediate M3 (73.6 g, yield 72.5%).

[0256] 1 H NMR (400 MHz, DMSO-d 6 ) δ7.27 (s, 1H), 7.25-7.22 (d, J = 12, 1H), 2.37 (s, 3H).

[0257] Example 4 Synthesis of Intermediate M4

[0258]

[0259] Steps:

[0260] Intermediate M3 (61.7 g, 0.24 mol) and (tributyltin)methanol (92.5 g, 0.29 mol) were dissolved in 1,4-dioxane (617 mL). 2 Xphos Pd G2 (9.4 g, 12 mmol) was added under protection, and the temperature was raised to 95±5°C for overnight reaction. TLC was monitored until the reaction was complete, and the reaction system was concentrated to dryness. 1553 ml of methyl tert-butyl ether was added to the residue and refluxed for 1 h. After cooling to room temperature, it was filtered and dried under reduced pressure to obtain intermediate M4 (40.16 g, yield 80.4%).

[0261] 1 H NMR (400 MHz, DMSO-d 6 ) δ12.47 (br.s, 1H), 7.08 (s, 1H), 7.04-7.00 (d, J=16, 1H), 5.48 (br.s, 1H), 4.56 (s, 2H), 2.41 (s, 3H).

[0262] Example 5 Synthesis of Intermediate M5

[0263]

[0264] Steps:

[0265] Under nitrogen protection, add intermediate M4 (1.0 g, 4.8 mmol), DIPEA (1.24 g, 9.6 mmol), DMAP (30 mg, 0.24 mmol) to DMF (5 mL), stir and cool to 0 ° C to obtain a reaction solution. Take p-toluenesulfonyl chloride (1.37 g, 7.2 mmol) and dissolve it in DMF (5 mL) and drop it into the reaction solution. After the drop is completed, warm to room temperature for reaction. Monitor by TLC until the reaction is complete. Dilute with water and extract with ethyl acetate three times. Combine the organic phases and wash three times with 10% brine. Separate the liquid, dry the organic phase, filter, concentrate under reduced pressure, and column chromatograph to obtain intermediate M5 (952 mg, yield 54.7%).

[0266] 1 H NMR (400 MHz, DMSO-d 6 +D 2 O) δ 8.10 (s, 1H), 8.08 (s, 1H), 7.62-7.56 (m, 4H), 4.72 (s, 2H), 2.64 (s, 3H), 2.45 (s, 3H).

[0267] Example 6 Synthesis of Intermediate M5

[0268]

[0269] Steps:

[0270] At 0°C, intermediate M4 (1.7 g, 8.2 mmol) and triphenylphosphine (6.4 g, 24.6 mmol) were dissolved in dichloromethane (50 mL) and stirred for 0.5 h. 2 Carbon tetrabromide (8.2 g, 24.6 mmol) was added under protection, and the reaction was allowed to proceed overnight at 0°C. TLC was used to monitor the reaction until it was complete, and the reaction system was concentrated to obtain a crude compound. The crude product was chromatographed on a silica gel column to obtain intermediate M5 (1.8 g, yield 81.4%).

[0271] 1 H NMR (400MHz, DMSO-d6) δ12.54(s,1H),7.21(dd,J=10.8,1.6Hz,1H),7.14(s,1H),4.77(s,2H),2.42(s,3H).

[0272] Example 7 Synthesis of Intermediate M6

[0273]

[0274] Steps:

[0275] Dissolve compound SM2 (20 g, 92.6 mmol), DIPEA (35.9 g, 277.7 mmol), and HATU (52.8 g, 138.9 mmol) in dichloromethane (200 mL), stir and cool to 0°C, add methylamine hydrochloride (12.5 g, 185 mmol). After addition, warm to room temperature for reaction. Monitor by TLC until the reaction is complete, add 300 ml of water, separate the liquids, and wash the organic phase with purified water, 10% potassium carbonate aqueous solution, and 10% saline water once. Separate the liquids, dry the organic phase, filter, concentrate under reduced pressure, and column chromatograph to obtain intermediate M6 (14.1 g, yield 66.5%).

[0276] 1 H NMR (400 MHz, CDCl 3 ) δ7.98-7.90 (m, 3H), 3.05-3.04 (d, J=4, 3H), 2.69 (s, 3H).

[0277] Example 8 Synthesis of Intermediate M7

[0278]

[0279] Steps:

[0280] Intermediate M6 (13.7 g, 59.8 mmol), tert-butyl piperazine-1-carboxylate (22.3 g, 119.6 mmol), cesium carbonate (58.5 g, 179.4 mmol) were added to 280 ml of toluene, replaced with nitrogen, tri(dibenzylideneacetone)dipalladium (2.7 g, 3 mmol), R-(+)-1,1'-binaphthyl-2,2'-bisdiphenylphosphine (3.7 g, 6 mmol) were added, replaced with nitrogen again, heated to 95 ° C and stirred. After TLC monitoring, 1 L of water was added to the reaction system after the reaction was complete, and extracted twice with ethyl acetate. The organic phases were combined and washed with water, 5% sodium dihydrogen phosphate aqueous solution, and 10% sodium chloride aqueous solution in turn. The organic phase was separated, dried, filtered, and concentrated under reduced pressure. The obtained solid was added to 700 ml of ethyl acetate, 24 g of hydrochloric acid (about 37%) was added, and stirred at room temperature. After the reaction was complete, 500 ml of water was added for separation under TLC monitoring. The aqueous phase was washed twice with ethyl acetate (200 ml*2). After separation, the aqueous phase was adjusted to weak alkalinity with 10% potassium carbonate aqueous solution. The organic phases were combined, washed with water, dried, filtered, and concentrated under reduced pressure to obtain intermediate M7 (12.6 g, yield 90%).

[0281] 1 H NMR (400 MHz, CDCl 3 +D 2 O) δ7.91-7.89 (d, J=8,1H), 7.27-7.25 (d, J=8,1H), 2.98-2.95 (m, 4H), 2.93 (s, 3H), 2.87-2.84 (m, 3H), 2.45 (s, 3H).

[0282] Example 9 Synthesis of Compound 8

[0283]

[0284] Steps:

[0285] Intermediate M5 (200 mg, 0.55 mmol), intermediate M7 (110 mg, 0.47 mmol), potassium carbonate (100 mg, 0.72 mmol) were added to 2 ml of DMF, heated to 80 °C and stirred. TLC was monitored until the reaction was complete. 2 ml of water was added to the reaction system, the temperature was lowered to 25 °C, and filtered. After column chromatography, compound 8 (170 mg, 85.2%) was obtained.

[0286] 1 H NMR (400 MHz, DMSO-d 6 )δ12.43(s,1H),8.42(q,J=4.8Hz,1H),7.80(d,J=8.4Hz,1H),7.49(d,J=8.4Hz,1H),7.14–7.05(m,2H ),3.62(s,2H),3.03–2.90(m,4H),2.80(d,J=4.8Hz,3H),2.63–2.54(m,4H),2.49(s,3H),2.41(s,3H).

[0287] Example 9 Synthesis of Compound 8

[0288]

[0289] Steps:

[0290] Intermediate M5 (6.00 g, 22.13 mmol) and intermediate M7 (6.24 g, 26.56 mmol) were added to 60 mL of 1,4-dioxane, stirring was started, N,N-diisopropylethylamine (5.72 g, 44.26 mmol) was added, and the temperature was raised to 45°C with stirring. TLC was monitored until the reaction was complete, the reaction system was cooled to 15°C, stirred for 2 h at this temperature, filtered, the filter cake was slurried with 180 mL of water at 20°C for 2 h, filtered, and the filter cake was dried under reduced pressure at 50°C and vacuum degree ≤-0.080 MPa to constant weight to obtain compound 8 (8.70 g, 92.60%).

[0291] 1 H NMR (400 MHz, DMSO-d 6 )δ12.44(s,1H),8.43(q,J=4.8Hz,1H),7.80(d,J=8.2Hz,1H),7.48(d,J=8.3Hz,1H),7.15–7.02(m,2H),3 .62(s,2H),2.96(t,J=4.8Hz,4H),2.82(d,J=4.9Hz,3H),2.59(t,J=4.7Hz,4H),2.49(s,3H),2.42(s,3H).

[0292] Pharmacological test evaluation

[0293] Test Example 1: PARP1 and PARP2 enzyme activity test

[0294] 1. Purpose of the test

[0295] This test is used to evaluate the effect of the test compound on PARP1 and PARP2 enzyme activities, and the inhibition rate is used to calculate the IC of the test compound on PARP1 and PARP2 enzymes. 50 value.

[0296] 2. Test materials

[0297] 2.1 Compounds:

[0298] Test drugs: Compounds AZD5305 and AZD9574 were purchased from Chengdu Dingdang Times Pharmaceutical Technology Co., Ltd.;

[0299] Test drug: Compound 8 of the present application.

[0300] 2.2 Test reagents and instruments

[0301] PARP1 Chemiluminescent assay kit, BPS, 80551;

[0302] PARP2 Chemiluminescent assay kit, BPS, 80552;

[0303] PBS, In house, 20210819;

[0304] Tween-20, Sigma, P9416;

[0305] 96-well polypropylene plate, Nunc, 249944;

[0306] Centrifuge, Xiangyi, TDZ5-WS;

[0307] Plate reader, BMG, PHERAstarFSX.

[0308] 3. Test methods

[0309] 3.1 Compound configuration

[0310] Preparation of 10 mM compound stock solution: Dissolve the compound powder in 100% DMSO to prepare 10 mM compound stock solution.

[0311] 3.2 Enzyme reaction process

[0312] (1) Dilute 5x histone mixture with PBS at a ratio of 1:5, add 50 μL to each well, and incubate at 4°C overnight.

[0313] (2) Wash three times with 200 μL PBST buffer and incubate in Blocking buffer for 90 min.

[0314] (3) Wash three times with 200 μL PBST buffer.

[0315] (4) Add 5 μL of inhibitor solution to each well.

[0316] (5) Add 20 μL of diluted PARP to each well.

[0317] (6) Add 25 μL of biotinylated substrate to each well and incubate at room temperature for 1 h.

[0318] (7) Wash the wells three times with 200 μL PBST buffer and dry with a clean paper towel.

[0319] (8) Add 50 μL of diluted Streptavidin-HRP to each well and incubate for 30 minutes.

[0320] (9) Wash three times with 200 μL PBST buffer.

[0321] (10) Add 100 μL of the mixture of ELISAECL substrate A and ELISAECL substrate B to each well.

[0322] (11) Read the chemiluminescent signal value immediately.

[0323] 3.3 Data Analysis

[0324] Calculation formula

[0325] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*Hillslope))

[0326] The logarithmic value of the compound concentration is used as the X-axis, the chemiluminescence signal is used as the Y-axis, the top and bottom are platforms with the same unit as Y, LogIC50 has the same logarithmic unit as X, Hillslop is the Slop factor or Hill slope, and the IC is fitted by a nonlinear regression equation. 50 value.

[0327] 4. Test results

[0328] The enzymatic inhibitory activity of the compounds of the present application on PARP1 and PARP2 was determined according to the above method. The results are shown in Table 2.

[0329] Table 2 Enzyme inhibitory activity of compounds against PARP1 and PARP2

[0330]

[0331] 5. Test conclusion:

[0332] From the test data of the enzymatic inhibitory activity of the compounds on PARP1 and PARP2 in Table 2, it can be seen that the compounds of the present application have strong inhibitory activity on PARP1 at the enzymatic level, and weak inhibitory effect on PARP2, and the enzymatic selectivity of some compounds for PARP1 and PARP2 is comparable to that of the positive control AZD9574, which has obvious advantages over the compound AZD5305 and can improve the blood toxicity caused by PARP2.

[0333] The above is only an implementation method of the present application, and its description is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present application. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present application, which all belong to the protection scope of the present application.

Claims

1. An intermediate M5; The X is selected from OTs, OMs, OTf, Cl, Br, I, p-nitrobenzenesulfonyloxy, 2,5-dichlorobenzenesulfonyloxy or 4-chlorobenzenesulfonyloxy; Preferably, X is selected from OTs, OMs, OTf, Cl, Br, I; X is preferably Br or X is preferably OTs.

2. A method for preparing the intermediate M5 according to claim 1, comprising: The intermediate M4 reacts with a sulfonylating agent or a halogenating agent to obtain the intermediate M5; Preferably, the sulfonylating agent is selected from methanesulfonyl chloride, phenylmethanesulfonyl chloride, p-toluenesulfonyl chloride, trifluoromethanesulfonyl chloride, trifluoromethanesulfonic anhydride, p-nitrobenzenesulfonyl chloride, 2,5-dichlorobenzenesulfonyl chloride and 4-chlorobenzenesulfonyl chloride, preferably p-toluenesulfonyl chloride; Preferably, the halogenating agent is selected from halogen, hydrogen halide, carbon tetrahalide, sulfoxide halide, phosphorus halide, triphenylphosphine, organic phosphine halide, N-haloamide and halogenated salt, preferably selected from carbon tetrabromide, chlorine, thionyl chloride, oxalyl chloride, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, hydrochloric acid, hydrogen chloride, triphenylphosphine, triphenylphosphine chloride, bromine, phosphorus tribromide, N-bromosuccinimide, triphenylphosphine bromide, N-iodosuccinimide and elemental iodine, preferably carbon tetrabromide and / or triphenylphosphine; Preferably, the molar ratio of the intermediate M4 to the sulfonylating agent is 1:1-5, preferably 1:1.5; Preferably, the molar ratio of the intermediate M4 to the halogenating agent is 1:1-10, preferably 1:3 or 1:6; Preferably, the step d) specifically comprises: Under alkaline conditions, the intermediate M4 reacts with the sulfonylating agent to obtain the intermediate M5; The base is preferably an inorganic base or an organic base; the inorganic base is preferably an alkali metal salt; the alkali metal salt is preferably one or more of alkali metal hydroxides, alkali metal carbonates, alkali metal bicarbonates and alkali metal phosphates; the organic base is preferably one or more of alkali metal salts of alcohols, lithium amide compounds, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), triethylamine, 4-dimethylaminopyridine and 1,8-diazabicyclo[5.4.0]undec-7-ene; the alkali metal salts of alcohols are preferably one or more of sodium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide; the lithium amide compound is preferably lithium diisopropylamide (LDA) or lithium hexamethyldisilazide (LiHMDS); The base is more preferably an organic base, and further preferably N,N-diisopropylethylamine (DIPEA) and / or 4-dimethylaminopyridine (DMAP); Preferably, the base is N,N-diisopropylethylamine (DIPEA) and 4-dimethylaminopyridine (DMAP); the molar ratio of the intermediate M4 to the N,N-diisopropylethylamine (DIPEA) is 1:1-5, preferably 1:2; the molar ratio of the intermediate M4 to 4-dimethylaminopyridine (DMAP) is 1:0.01-0.2, preferably 1:0.05; Preferably, the step d) specifically comprises: reacting the intermediate M4 with the sulfonylating agent or the halogenating agent in a solvent D to obtain the intermediate M5; Preferably, the step d) specifically comprises: reacting the intermediate M4 with p-toluenesulfonyl chloride in a solvent D to obtain the intermediate M5; The solvent D is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; for example, the ether solvent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether and tetrahydropyran; the halogenated hydrocarbon solvent is one or more of dichloromethane, chloroform, carbon tetrachloride and dichloroethane; the aromatic hydrocarbon solvent is one or more of toluene, benzene, xylene, trimethylbenzene and chlorobenzene; the ester solvent is ethyl acetate; The solvent D is more preferably dimethylformamide (DMF) or dichloromethane; Preferably, the preparation method of step d) specifically comprises: under alkaline conditions, the intermediate M4 and the sulfonylating agent react in solvent D to obtain the intermediate M5; Preferably, the preparation method in step d) specifically comprises: reacting the intermediate M4 with the halogenating agent in a solvent D to obtain the intermediate M5; Preferably, the reaction temperature of step d) is 0°C-100°C; the intermediate M4 reacts with the sulfonylating agent, and the reaction temperature is preferably room temperature; the intermediate M4 reacts with the halogenating agent, and the reaction temperature is preferably 0°C; Preferably, the reaction temperature of step d) is 0°C-100°C, preferably room temperature.

3. The preparation method according to claim 2, characterized in that: X is OTs; preferably, the step d) specifically comprises: at 0°C, 8.2 mmol of intermediate M4 and 24.6 mmol of triphenylphosphine are dissolved in 50 mL of dichloromethane, stirred for 0.5 h, 24.6 mmol of carbon tetrabromide is added under N2 protection, and the reaction is carried out overnight at 0°C to obtain intermediate M5; more preferably, the step d) specifically comprises: at 0°C, 8.2 mmol of intermediate M4 and 24.6 mmol of triphenylphosphine are dissolved in 50 mL of dichloromethane, stirred for 0.5 h, 24.6 mmol of carbon tetrabromide is added under nitrogen protection, the reaction is carried out overnight at 0°C, the reaction is complete, the reaction system is concentrated to obtain a crude compound, and the obtained crude product is subjected to silica gel column chromatography to obtain intermediate M5; or, X is Br; preferably, the step d) specifically comprises: under nitrogen protection, 4.8mmol intermediate M4, 9.6mmol DIPEA, and 0.24mmol DMAP are added to 5mL DMF, stirred and cooled to 0°C; 7.2mmol p-toluenesulfonyl chloride is dissolved in 5mL DMF, and the mixture is added dropwise to the reaction solution, and the mixture is heated to room temperature for reaction; more preferably, the step d) specifically comprises: under nitrogen protection, 4.8mmol intermediate M4, 9.6mmol DIPEA, and 0.24mmol DMAP are added to 5mL DMF, stirred and cooled to 0°C to obtain a reaction solution; 7.2mmol p-toluenesulfonyl chloride is dissolved in 5mL DMF, and the mixture is added dropwise to the reaction solution; the mixture is heated to room temperature for reaction; after the reaction is complete, the mixture is diluted with water, extracted three times with ethyl acetate, the organic phases are combined, and washed three times with 10% brine; the organic phases are separated, dried, filtered, concentrated under reduced pressure, and subjected to column chromatography to obtain the intermediate M5.

4. The preparation method according to claim 2, characterized in that: Also includes: Step e) reacting the intermediate M3 with (tributyltin)methanol to obtain the intermediate M4; Preferably, the molar ratio of the intermediate M3 to the (tributyltin)methanol is 1:1-3, preferably 1:1.2; Preferably, the step e) specifically comprises: reacting the intermediate M3 and the (tributyltin)methanol in a solvent E to obtain the intermediate M4; The solvent E is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; the solvent E is more preferably 1,4-dioxane; Preferably, the mass volume ratio of the intermediate M3 to the solvent E is 1g:5-20ml, preferably 1g:10ml; Preferably, the preparation method in step e) specifically comprises: under the condition of catalyst E, the intermediate M3 and the (tributyltin)methanol react to obtain the intermediate M4; The catalyst E is preferably a metal catalyst, and the metal catalyst is preferably any one of a palladium catalyst, a cobalt catalyst and a nickel catalyst; The catalyst E is more preferably chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (Xphos Pd G2); Preferably, the molar ratio of the intermediate M3 to the catalyst E is 5-30:1, preferably 20:1; Preferably, the preparation method in step e) specifically comprises: under the condition of catalyst E, the intermediate M3 and the (tributyltin)methanol react in solvent E to obtain the intermediate M4; Preferably, the reaction temperature of step e) is 60°C-120°C, preferably 90°C-100°C; Preferably, the reaction time of step e) is 12h-16h; More preferably, the step e) specifically comprises: dissolving 0.24 mol of intermediate M3 and 0.29 mol of (tributyltin) methanol in 617 mL of 1,4-dioxane, adding 12 mmol of Xphos Pd G2 under N2 protection, heating to 95±5°C and reacting overnight; Most preferably, the step e) is specifically as follows: 0.24 mol of intermediate M3 and 0.29 mol (tributyltin) methanol are dissolved in 617 mL of 1,4-dioxane, 12 mmol of Xphos Pd G2 is added under N2 protection, the temperature is raised to 95±5°C and the reaction is carried out overnight. After the reaction is complete, the reaction system is concentrated to dryness, 1553 ml of methyl tert-butyl ether is added to the residue, the mixture is refluxed and stirred for 1 h, the temperature is cooled to room temperature and then filtered, and the mixture is dried under reduced pressure to obtain intermediate M4.

5. The preparation method according to claim 4, characterized in that: Also includes: Dehydrogenation reaction of the intermediate M2 to obtain the intermediate M3; Preferably, the step f) specifically comprises: dehydrogenating the intermediate M2 in a solvent F to obtain the intermediate M3; The solvent F is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; the solvent F is more preferably 1,4-dioxane; Preferably, the mass to volume ratio of the intermediate M2 and the solvent F is 1 g:5-30 ml, preferably 1 g:20 ml; Preferably, the preparation method in step f) specifically comprises: under oxidizing conditions, dehydrogenating the intermediate M2 to obtain the intermediate M3; Preferably, the molar ratio of the intermediate M2 to the oxidant is 1:1-4, preferably 1:1.175; The oxidant is preferably 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) or manganese dioxide, preferably 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ); Preferably, the reaction temperature of step f) is 0-100°C, preferably room temperature; Preferably, the reaction time of step f) is 12h-16h; More preferably, the preparation method of step f) specifically comprises: dissolving 0.40 mol of intermediate M2 in 1020 mL of 1,4-dioxane, cooling to 0° C. after nitrogen substitution, dissolving 0.47 mol of DDQ in 1020 mL of 1,4-dioxane, slowly dripping into the reaction system, and reacting at room temperature overnight; Most preferably, the preparation method of step f) is specifically as follows: 0.40 mol of intermediate M2 is dissolved in 1020 mL of 1,4-dioxane, the temperature is lowered to 0°C after nitrogen replacement, 0.47 mol of DDQ is dissolved in 1020 mL of 1,4-dioxane, and the mixture is slowly dripped into the reaction system; after the dripping, the mixture is reacted at room temperature overnight, and after the reaction is complete, the mixture is cooled to 5±5°C, 3 L of saturated sodium bicarbonate aqueous solution is dripped, the mixture is stirred for 1 hour at this temperature, and then filtered to obtain a solid; the obtained solid is subjected to a hot pulping process, the hot pulping process is repeated 5 times, the solid is filtered, and the solid is dried under reduced pressure to obtain an intermediate M3.

6. The preparation method according to claim 5, characterized in that: Also includes: Step g) reacting the intermediate M1 to obtain the intermediate M2; Preferably, the step g) specifically comprises: reacting the intermediate M1 in a solvent G to obtain the intermediate M2; The solvent G is preferably a mixed solvent of glacial acetic acid and water; the volume ratio of glacial acetic acid to water is preferably 5-12:1, preferably 9:1; The mass volume ratio of the intermediate M1 and the solvent G is 1g:5-20ml, preferably 1g:10ml; Preferably, the step g) specifically comprises: under the condition of catalyst G, the intermediate M1 reacts to obtain the intermediate M2; The catalyst G is preferably zinc powder, palladium carbon, iron powder, tin dichloride, ferrous sulfate, sodium dithionite, and more preferably zinc powder; The molar ratio of the solvent G to the catalyst G is 1:3-10, preferably 1:5; Preferably, the step g) specifically comprises: under the condition of catalyst G, the intermediate M1 reacts in solvent G to obtain the intermediate M2; Preferably, the reaction temperature of step g) is 50-100°C, preferably 90°C to 100°C; Preferably, the reaction time of step g) is 12h-16h; More preferably, the step g) specifically comprises: dissolving the 0.43 mol intermediate M1 in a mixed solvent of 1242 mL glacial acetic acid and 138 mL water, cooling the reaction system to 0° C., adding 2.15 mol zinc powder in batches, replacing nitrogen after the addition, raising the reaction system to 95±5° C. and reacting overnight to obtain the intermediate M2; Most preferably, the step g) is specifically 0.43 mol of intermediate M1 (138.0 g, 0.43 mol) is dissolved in glacial acetic acid / water (1242 mL / 138 mL), the reaction system is cooled to 0°C, 2.15 mol of zinc powder is added, nitrogen is replaced after the addition, the reaction system is heated to 95±5°C and reacted overnight, the reaction is completed, the reaction system is cooled to room temperature and diluted with water, extracted three times with ethyl acetate, the organic phases are combined, and the organic phases are washed with saturated sodium bicarbonate aqueous solution until the aqueous phase is weakly alkaline, and the organic phase is concentrated to obtain intermediate M2.

7. The preparation method according to claim 6, characterized in that: Also includes: Step h) reacting the intermediate SM1 with alanine methyl ester hydrochloride to obtain the intermediate M1; Preferably, the molar ratio of the compound SM1 to the alanine methyl ester hydrochloride is 1:1-3, preferably 1:1.1; Preferably, the step h) specifically comprises: under alkaline conditions, reacting the compound SM1 with the alanine methyl ester hydrochloride to obtain the intermediate M1; The base is preferably an organic base; the organic base is preferably N,N-diisopropylethylamine (DIPEA), sodium tert-butoxide, tert-butylmagnesium chloride, triethylamine, pyridine or 4-dimethylaminopyridine (DMAP); The base is more preferably N,N-diisopropylethylamine (DIPEA); Preferably, the step h) specifically comprises: reacting the compound SM1 and the alanine methyl ester hydrochloride in a solvent H to obtain the intermediate M1; The solvent H is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; the solvent H is more preferably 1,4-dioxane; Preferably, the preparation method of step h) specifically comprises: under alkaline conditions, the compound SM1 and the alanine methyl ester hydrochloride; Preferably, the reaction temperature of step h) is 20°C-100°C, preferably 55°C-65°C; Preferably, the reaction time of step h) is 2-10 h, preferably 3 h; More preferably, the step h) specifically comprises: dissolving the 0.42 mol compound SM1 (100.0 g, 0.42 mol) and 0.46 mol alanine methyl ester hydrochloride in 140 mL 1,4-dioxane, adding 1.26 mol DIPEA, and heating the reaction system to 60±5° C. for 3 h to obtain the intermediate M1; Most preferably, the step h) is specifically as follows: 0.42 mol of compound SM1 and 0.46 mol of alanine methyl ester hydrochloride are dissolved in 140 mL of 1,4-dioxane, 1.26 mol of DIPEA is added, and the reaction system is heated to 60±5° C. for 3 hours after the addition is completed. After the reaction is complete, the reaction system is concentrated, diluted with water, extracted three times with ethyl acetate, the organic phases are combined, washed three times with 10% sodium chloride aqueous solution, and the organic phase is dried and concentrated to obtain the intermediate M1.

8. A method for preparing compound 8, comprising: Step a) reacting the intermediate M5 of claim 1 with the intermediate M7 to obtain the compound 8; Preferably, the molar ratio of the intermediate M5 to the intermediate M7 is 0.80:1-1:1.3, preferably 1:0.85 or 1:1.2; Preferably, the step a) specifically comprises: Under alkaline conditions, the intermediate M5 and the intermediate M7 react to obtain the compound 8; The base is preferably an inorganic base or an organic base; the inorganic base is preferably an alkali metal salt; the alkali metal salt is preferably one or more of alkali metal hydroxides, alkali metal carbonates, alkali metal bicarbonates and alkali metal phosphates; the organic base is preferably one or more of alkali metal salts of alcohols, lithium amide compounds, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), triethylamine, 4-dimethylaminopyridine and 1,8-diazabicyclo[5.4.0]undec-7-ene; the alkali metal salts of alcohols are preferably one or more of sodium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide; the lithium amide compound is preferably lithium diisopropylamide (LDA) or lithium hexamethyldisilazide (LiHMDS); The base is more preferably an inorganic base; further preferably potassium carbonate; The base is more preferably an organic base; further preferably N,N-diisopropylethylamine (DIPEA); Preferably, the step a) specifically comprises: reacting the intermediate M5 and the intermediate M7 in solvent A to obtain the compound 8; The solvent A is preferably one or more of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dimethylacetamide (DMAC), N-methylpyrrolidone (NMP), 1,4-dioxane, acetonitrile, ethanol, ether solvents, halogenated hydrocarbon solvents, aromatic hydrocarbon solvents and ester solvents; the solvent A is more preferably dimethylformamide (DMF); the solvent A is more preferably 1,4-dioxane; Preferably, the preparation method of step a) specifically comprises: under alkaline conditions, the intermediate M5 and the intermediate M7 react in solvent A to obtain the compound 8; Preferably, the reaction temperature of step a) is 40°C-110°C, preferably 45-80°C, more preferably 45°C or 80°C.

9. The preparation method according to claim 8, characterized in that: X is OTs; preferably, the step a) specifically comprises: under potassium carbonate conditions, the intermediate M5 and the intermediate M7 react to obtain the compound 8, and the molar ratio of the intermediate M5 to the intermediate M7 is 1:0.85; more preferably, the step a) specifically comprises adding 0.55 mmol of the intermediate M5, 0.47 mmol of the intermediate M7, and 0.72 mmol of potassium carbonate to 2 ml of DMF, heating to 80°C and stirring, and reacting completely; adding 2 ml of water to the reaction system, cooling to 25°C, and filtering; and obtaining compound 8 by column chromatography; or, X is Br; preferably, the step a) specifically comprises: adding 22.13 mmol intermediate M5 and 26.56 mmol intermediate M7 to 60 mL 1,4-dioxane, stirring, adding 44.26 mmol N, N-diisopropylethylamine, heating to 45°C to react to obtain the compound 8; more preferably, the step a) specifically comprises: adding 22.13 mmol intermediate M5 and 26.56 mmol intermediate M7 to 60 mL 1,4-dioxane, starting stirring, adding 44.26 mmol N, N-diisopropylethylamine, heating to 45°C and stirring, reacting completely, cooling the reaction system to 15°C, keeping warm and stirring for 2 hours, filtering, slurrying the filter cake with 180 mL water at 20°C for 2 hours, filtering, and drying the filter cake under reduced pressure at a temperature of 50°C and a vacuum degree of ≤-0.080 MPa to constant weight to obtain compound 8.

10. The preparation method according to claim 8, characterized in that: include: The intermediate SM1 reacts with alanine methyl ester hydrochloride to obtain the intermediate M1; The intermediate M1 is reacted to obtain the intermediate M2; The intermediate M2 is reacted to obtain the intermediate M3; The intermediate M3 reacts with (tributyltin)methanol to obtain the intermediate M4; The intermediate M4 reacts with a sulfonylating agent or a halogenating agent to obtain an intermediate M5; Compound SM2 reacts with methylamine hydrochloride to obtain intermediate M6; The intermediate M6 reacts with piperazine-1-carboxylic acid tert-butyl ester to obtain compound M7; The intermediate M5 and the intermediate M7 react to obtain the compound 8;

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