Oxime ether compounds, methods of making, pharmaceutical compositions, and uses thereof

Abstract

The application discloses an oxime ether compound, a preparation method, a pharmaceutical composition and application thereof. The compound structure is shown in formula (I), and the compound also comprises isomers, prodrugs, stable isotopically substituted compounds, pharmaceutically acceptable salts or mixtures thereof. The compound can effectively inhibit PRMT5 enzyme and the activity of MTAP deletion type cells, can exert PRMT5-MTA synergistic pharmacodynamics at the molecular level and the cell level, can be used for treating various tumors, has higher selectivity for MTAP deletion type cells, does not affect the activity of PRMT5 in wild type cells, is safer, and the preparation method is simple and easy to operate.

Description

Technical Field

[0001] This invention relates to an oxime ether compound, its preparation method, pharmaceutical composition, and application, and more particularly to an oxime ether compound with antitumor activity, its preparation method, pharmaceutical composition, and application. Background Technology

[0002] Protein arginine methyltransferases (PRMTs) are S-adenosylmethionine (SAM)-dependent methyltransferases that transfer methyl groups from SAM to the guanidino nitrogen atom of histone and non-histone arginine residues. Based on the methylated form of the resulting arginine, PRMTs are classified into type I (PRMT1, 2, 3, 4, 6, 8), type II (PRMT5, 9), and type III (PRMT7), with PRMT5 catalyzing the formation of monomethylarginine and symmetrical dimethylarginine. PRMTs play important roles in various cellular functions, influencing gene expression in signal transduction, mRNA splicing, and DNA repair through histone methylation. PRMT5 expression is upregulated in various human malignancies, including lung cancer, ovarian cancer, colorectal cancer, breast cancer, melanoma, leukemia, and malignant gliomas. Furthermore, PRMT5 has been identified as an anticancer target for mantle cell lymphoma and glioblastoma.

[0003] The gene encoding methionine phosphorylase (MTAP) is widely expressed in normal tissues. CDKN2A is one of the most frequently deleted tumor suppressor genes, and MTAP is similar to CDKN2A; in tumor cells, MTAP is often co-deleted with CDKN2A. MTAP can cleave methionine (MTA) to produce methionine and adenine salvage pathway precursor substrates. Therefore, MTAP-deficient cells have increased MTA levels. MTA competitively binds to PRMT5 with SAM, leading to decreased PRMT5 activity. Thus, MTAP-deficient cells are more sensitive to further reductions in PRMT5 enzyme activity.

[0004] First-generation PRMT5 inhibitors are mainly divided into SAM non-competitive and SAM competitive types. Representative drugs include GSK-3326595, JNJ-64619178, PF-06939999, PRT543, and PRT811. They all have non-selective inhibitory activity against PRMT5 and are associated with dose-limiting conditions such as thrombocytopenia, anemia, and neutropenia. They also have drawbacks such as large side effects and low therapeutic index, which limits the application of first-generation PRMT5 inhibitors. Summary of the Invention

[0005] Objectives of the Invention: The first objective of this invention is to provide an oxime ether compound; the second objective is to provide a method for preparing the compound; the third objective is to provide a pharmaceutical composition comprising the compound; and the fourth objective is to provide an application of the compound and the pharmaceutical composition thereof.

[0006] Technical solution: The oxime ether compounds of the present invention have the structure of formula (I), and further comprise isomers thereof, prodrugs, stable isotope-substituted compounds, pharmaceutically acceptable salts, or mixtures thereof:

[0007]

[0008] in:

[0009] R 1 Selected from the substituted C6~C 10 Aryl, substituted C5-C9 heteroaryl, substituted C3-C8 cycloalkyl, substituted C2-C6 heterocycloalkyl, substituted C4-C 12 Spirocyclic or substituted C4-C 12 Bridged cycloyl group, wherein the substituent is selected from at least one hydrogen, halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 fluoroalkoxy, C2-C6 heterocyclic alkyl, hydroxyl, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl or nitro, wherein the heteroaryl or heterocyclic alkyl group contains 1 to 3 heteroatoms selected from N, O, S;

[0010] R 2 or R 3 Each of the substituents is independently selected from hydrogen, substituted C1-C6 alkyl, or substituted C1-C6 haloalkyl, wherein the substituent is selected from at least one hydrogen, halogen, cyano, C3-C8 cycloalkyl, C2-C6 heterocycloalkyl, hydroxyl, amino, methylamino, dimethylamino, nitro, carboxyl, or methoxycarbonyl, and the heteroaryl or heterocycloalkyl contains 1 to 3 heteroatoms selected from N, O, and S;

[0011] R 4 Selected from hydrogen, substituted C1-C8 alkyl, substituted C1-C8 haloalkyl, substituted C6-C 10 Aryl, substituted C5-C9 heteroaryl, substituted C3-C8 cycloalkyl, substituted C4-C 12 Spirocyclic groups, substituted C4-C 12The substituent is selected from at least one hydrogen, halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C2-C6 heterocyclic alkyl, hydroxyl, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl, or nitro, and the heteroaryl or heterocyclic alkyl group contains 1 to 3 heteroatoms selected from N, O, and S.

[0012] Ring A is selected from substituted five-membered aromatic heterocycles or five-membered heterocyclic alkanes, wherein the substituent is selected from at least one hydrogen, halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, C2-C6 heterocyclic alkyl, C1-C6 alkoxy, C1-C6 acyl, hydroxyl, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl, or nitro, and the aromatic heterocycle, heterocyclic alkanes, or heterocyclic alkyl groups contain 1 to 3 heteroatoms selected from N, O, and S;

[0013] X 1 or X 2 Selected independently from CR 5 Or N, where R 5 It is selected from hydrogen, halogen, amino, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy.

[0014] Preferably, in the structure:

[0015] R 1 Selected from the substituted C6~C 10 The substituent is selected from at least one hydrogen, halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 fluoroalkoxy, C2-C6 heterocyclic alkyl, hydroxyl, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl, or nitro, and the heteroaryl or heterocyclic alkyl contains 1 to 3 heteroatoms selected from N, O, and S.

[0016] R 2 or R 3 Each is independently selected from hydrogen or methyl;

[0017] R 4 Selected from hydrogen, substituted C1-C6 alkyl, substituted C1-C6 haloalkyl, substituted C6-C 10 Aryl, substituted C5-C9 heteroaryl, substituted C3-C8 cycloalkyl, substituted C4-C 12Spirocyclic, substituted vinyl, substituted ethynyl or substituted allyl, wherein the substituent is selected from at least one hydrogen, halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C2-C6 heterocyclic alkyl, hydroxyl, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl or nitro, and the heteroaryl or heterocyclic alkyl contains 1 to 3 heteroatoms selected from N, O, S;

[0018] Ring A is selected from substituted five-membered heteroaromatic rings or five-membered heterocyclic alkanes, wherein the substituent is selected from at least one hydrogen, halogen, methyl, ethyl, isopropyl, cyclopropyl, trifluoromethyl, trifluoroethyl, methoxy, cyclopropylmethoxy, trifluoromethoxy, or 2,2-difluoroethyl, and the heteroaromatic ring or heterocyclic alkanes contain 1 to 3 heteroatoms selected from N, O, and S.

[0019] X 1 or X 2 Selected independently from CR 5 Or N, R 5 It is selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, trifluoromethyl, methoxy, trifluoromethoxy, or 2,2-difluoroethyl.

[0020] Preferably, in the structure:

[0021] R 1 The substituent is selected from substituted benzene rings, substituted pyridines, substituted pyridazines, substituted pyrazines, substituted furans, substituted thiophenes, substituted oxazoles, substituted pyrazoles, substituted imidazoles, substituted thiazoles, substituted pyrimidines, substituted naphthyl rings, substituted quinolines, substituted indoles, substituted indazoles, substituted cyclopropyl, substituted cyclobutyl, substituted cyclopentyl, substituted cyclohexyl, substituted piperazine, substituted piperidine, substituted pyrrole, or substituted tetrahydrofuran, wherein the substituent is selected from at least one hydrogen, fluorine, chlorine, bromine, cyano, methyl, ethyl, trifluoromethyl, difluoromethyl, cyclopropyl, methoxy, trifluoromethoxy, cyclopropoxy, morpholino, piperidinyl, tetrahydropyrrolidinyl, 2-(N,N-dimethylamino)ethyl, 2-(N,N-diethylamino)ethyl, hydroxyl, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl, or nitro.

[0022] R 2 or R 3 Each is independently selected from hydrogen or methyl;

[0023] R 4 Selected from hydrogen, substituted C1-C6 alkyl, substituted C1-C6 haloalkyl, substituted C6-C 10 Aryl, substituted C5-C9 heteroaryl, substituted C3-C8 cycloalkyl, substituted C4-C 12Spirocyclic or substituted allyl, wherein the substituent is selected from at least one hydrogen, fluorine, chlorine, bromine, cyano, methyl, ethyl, trifluoromethyl, difluoromethyl, cyclopropyl, methoxy, cyclopropylmethoxy, trifluoromethoxy, acrylidine, tetrahydropyrrolyl, morpholino, piperidinyl, piperazine, 2-(N,N-dimethylamino)ethyl, 2-(N,N-diethylamino)ethyl, hydroxyl, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl, or nitro, and the heteroaryl group comprises 1 to 3 heteroatoms selected from N, O, and S;

[0024] Ring A is selected from substituted pyrroles, pyrazoles, isoxazoles, thiophenes, thiazoles, furans, dihydropyrroles, or dihydrofurans, wherein the substituent is selected from at least one hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, or trifluoromethoxy.

[0025] X 1 or X 2 Selected independently from CR 5 Or N, R 5 It is selected from hydrogen, methyl, ethyl, trifluoromethyl, methoxy, trifluoromethoxy or 2,2-difluoroethyl.

[0026] Preferably, in the structure:

[0027] R 1 Selected from

[0028] R 2 or R 3 Each is independently selected from hydrogen or methyl;

[0029] R 4 Selected from -CH3, -CH2CH3, -CH(CH3)2, -C(CH3)3, -CH2CH2OH, -CH2CH2CN, -CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -CH2CH2OCH3, -CH2CH=CH2,

[0030] Ring A is selected from

[0031] X 1 or X 2 Each is independently selected from CH, CF, C-Cl, C-Br, or N.

[0032] Specifically, the oxime ether compound is selected from any of the following compounds:

[0033]

[0034]

[0035]

[0036]

[0037]

[0038]

[0039]

[0040] This invention designs a series of MTA-synergistic PRMT5 inhibitors that can selectively inhibit PRMT5 activity in MTAP-deficient cells while preserving PRMT5 activity in MTAP-wild-type cells, making them safer and more effective.

[0041] Preferably, the pharmaceutically acceptable salt is a salt formed by the compound with any of the following acids or bases: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or ferulic acid.

[0042] The method for preparing oxime ether compounds according to the present invention includes the following steps:

[0043] When R 3 When it is hydrogen,

[0044] (1) Compound 1 was reacted with pinacol diboronic acid ester to prepare compound 2;

[0045] (2) Compound 3 was halogenated and then coupled with compound 2 to obtain compound 5;

[0046] (3) Compound 5 was cyclized, hydrolyzed, and halogenated to obtain compound 8;

[0047] (4) Compound 9 was condensed and reduced with an amino ether salt, and then acylated with compound 8 to obtain compound I;

[0048]

[0049] Among them, R 1 R 2 R 4 X 1 X 2 The definition is as stated above;

[0050] The corresponding acid is used to form a salt with compound I prepared by the above method to obtain a pharmaceutically acceptable salt of the oxime ether compound.

[0051] Specifically, in step (1), compound 2 is prepared by reacting compound 1 with pinacol diboronate. The catalyst used is selected from [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (Pd(dppf)Cl2), tetra(triphenylphosphine)palladium (Pd(PPh3)4), bis(triphenylphosphine)palladium dichloride (Pd(PPh3)2Cl2) or palladium acetate (Pd(OAc)2), preferably Pd(dppf)Cl2; the solvent used is selected from dichloromethane, tetrahydrofuran, toluene, 1,4-dioxane, ethyl acetate, acetone, N,N-dimethylformamide or a mixture of any two, preferably 1,4-dioxane; the base used is selected from potassium hydroxide, potassium carbonate, potassium acetate, sodium carbonate, preferably potassium acetate; the reaction temperature is selected from 80 to 120°C, preferably 100°C.

[0052] In step (2), compound 4 is prepared from compound 3 via the Sandmeier reaction. The diazotizing agent used is selected from methyl nitrite, ethyl nitrite, amyl nitrite, isoamyl nitrite, isobutyl nitrite, tert-butyl nitrite, and isopropyl nitrite, preferably isoamyl nitrite. The solvent and iodizing agent used are selected from iodomethane, diiodomethane, triiodomethane, iodoethane, diiodoethane, or a mixture of any two, preferably diiodomethane. The reaction temperature is selected from 60 to 100°C, preferably 80°C.

[0053] Compound 5 is prepared by reacting compound 2 with compound 4. The solvent used is selected from a mixture of any two of toluene, 1,4-dioxane, N,N-dimethylformamide, n-butanol, and water, preferably a mixture of 1,4-dioxane and water. The base used is selected from potassium hydroxide, potassium carbonate, cesium carbonate, potassium acetate, sodium carbonate, and sodium bicarbonate, preferably cesium carbonate. The catalyst used is selected from [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (Pd(dppf)Cl2), tetrahydropalmatine, and tetrahydropalmatine. (Triphenylphosphine)palladium (Pd(PPh3)4), bis(triphenylphosphine)palladium dichloride (Pd(PPh3)2Cl2), palladium acetate (Pd(OAc)2), and chloro(2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (SPhospdG2), preferably tetra(triphenylphosphine)palladium (Pd(PPh3)4); the reaction temperature is selected from 60 to 100 °C, preferably 80 °C.

[0054] In step (3), compound 6 is prepared from compound 5 by aromatization reaction. The solvent used is selected from anhydrous toluene, anhydrous tetrahydrofuran, anhydrous 1,4-dioxane, anhydrous N,N-dimethylformamide or a mixture of any two solvents, preferably anhydrous toluene; the base used is selected from sodium amide, potassium hydroxide, potassium carbonate, potassium acetate, sodium carbonate, preferably sodium amide; the reaction temperature is selected from 80 to 120°C, preferably 100°C.

[0055] Compound 7 is prepared by hydrolysis of compound 6. The base used is selected from lithium hydroxide, sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, preferably lithium hydroxide. The solvent used is selected from methanol, ethanol, tetrahydrofuran, dioxane, water, or a mixture thereof, preferably a mixture of methanol:tetrahydrofuran:water = 1:1:1. The reaction temperature is selected from 50 to 90°C, preferably 60°C.

[0056] Compound 8 was prepared by acylation of compound 7. The chlorinating agent was selected from thionyl chloride, phosphorus trichloride, phosphorus pentachloride, and oxalyl chloride, with oxalyl chloride being preferred.

[0057] In step (4), the intermediate oxime is prepared by reacting compound 9 with compound 10. The solvent used is selected from dichloromethane, tetrahydrofuran, 1,4-dioxane, ethyl acetate, N,N-dimethylformamide, or a mixture of any two, preferably dichloromethane; the base used is selected from triethylamine, pyridine, N,N-diisopropylethylamine, 4-dimethylaminopyridine, potassium carbonate, sodium carbonate, cesium carbonate, or sodium acetate, preferably pyridine. Compound 11 is prepared from the intermediate oxime by reduction reaction. The reducing agent used is selected from sodium borohydride, sodium triacetoxyborohydride, or sodium cyanoborohydride, preferably sodium cyanoborohydride; the solvent used is selected from methanol, ethanol, dioxane, tetrahydrofuran, glacial acetic acid, or a mixture of any two, preferably glacial acetic acid.

[0058] Final product I was prepared by reacting compound 8 with compound 11. The solvent used was selected from dichloromethane, ethyl acetate, tetrahydrofuran, and 1,4-dioxane, with tetrahydrofuran being preferred. The base used was selected from triethylamine, pyridine, and N,N-diisopropylethylamine, with N,N-diisopropylethylamine being preferred.

[0059] The pharmaceutical composition of this invention comprises the oxime ether compound and a pharmaceutically acceptable carrier. Common pharmaceutical excipients such as flavorings, sweeteners, liquid / solid fillers, and diluents can be added to formulate common pharmaceutical preparations, such as tablets, capsules, syrups, suspensions, and injections.

[0060] The oxime ether compounds and their pharmaceutical compositions described in this invention are used to prepare PRMT5 inhibitor drugs, specifically as anti-tumor drugs, and particularly as drugs for treating breast cancer, colorectal cancer, hematologic malignancies, etc.

[0061] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0062] This oxime ether compound can effectively inhibit the activity of PRMT5 enzyme and MTAP-deficient cells (IC50). 50 Most of the concentrations reach nanomolar levels, with the optimal concentration below 10 nM. PRMT5 and MTA can exert synergistic effects at both the molecular and cellular levels, and can be used to treat various tumors. At the same time, it has higher selectivity for MTAP-deficient cells, does not affect the activity of PRMT5 in wild-type cells, and is safer. In addition, the compound preparation method is simple and easy to operate. Detailed Implementation

[0063] The technical solution of the present invention will be further described below with reference to the embodiments.

[0064] Example 1: 4-amino-N-((5-bromopyridin-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-1: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxoborheptacyclopentan-2-yl)benzoate (2)

[0065] To a 250 mL round-bottom flask, methyl 4-amino-3-bromobenzoate 1 (4.00 g, 17.4 mmol), pinacol diborate (8.83 g, 34.8 mmol), and 60 mL of 1,4-dioxane were added sequentially. Potassium acetate (5.12 g, 52.2 mmol) and Pd(dppf)Cl2 (1.42 g, 1.74 mmol) were then added under stirring at room temperature. The reaction system was refluxed at 100 °C for 18 h under N2 protection. TLC (VL) was performed. 石油醚 :V 乙酸乙酯 The reaction was monitored to ensure complete reaction (1:1 ratio), cooled to room temperature, and filtered. The filter cake was washed with dichloromethane, and the filtrate was concentrated to obtain a semi-solid residue. This residue was then dissolved in a small amount of dichloromethane, filtered, and the filter cake was collected. The filtrate was concentrated, and the operation was repeated once. The two filter cakes were combined and dried to obtain 3.00 g of a grayish-white solid, with a yield of 62.2%.

[0066] 1H NMR (300MHz, CDCl3-d) δ (ppm): 8.30 (d, J = 2.2Hz, 1H), 7.87 (dd, J = 8.6, 2.2Hz, 1H), 6.55 (d, J = 8.6Hz, 1H), 3.84 (s, 3H), 1.34 (s, 12H).

[0067] Synthesis of 5-iodo-1-methyl-1H-pyrazole-4-onitrile (4-1)

[0068] Add 5-amino-1-methyl-1H-pyrazole-4-onitrile 3-1 (5.00 g, 40.9 mmol) and 90 mL of CH2I2 sequentially to a 250 mL three-necked flask. Heat to 80 °C, then add amyl nitrite (11.0 mL, 81.9 mmol) dropwise. After the addition is complete, continue heating at 80 °C for 1 h. TLC (V 石油醚 :V 乙酸乙酯 =10:1) Monitor the reaction to ensure it is complete, cool to room temperature, extract the reaction solution with concentrated HCl (30 mL × 2), combine the concentrated HCl layers, adjust the pH of the aqueous layer to 1 with saturated NaOH aqueous solution, then extract with ethyl acetate (30 mL × 2), combine the ethyl acetate layers, remove the solvent under reduced pressure, and precipitate the residue by column chromatography (V... 石油醚 :V 乙酸乙酯 Purification was performed using a ratio of 20:1 to obtain 3.00 g of a white solid, with a yield of 31.5%.

[0069] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 7.81 (s, 1H), 3.99 (s, 3H).

[0070] Synthesis of methyl 4-amino-3-(4-cyano-1-methyl-1H-pyrazole-5-yl)benzoate (5-1)

[0071] To a 250 mL round-bottom flask, add intermediate 2 (3.92 mg, 14.2 mmol), intermediate 4-1 (3.00 g, 12.9 mmol), Pd(PPh3)4 (744 mg, 0.644 mmol), cesium carbonate (12.6 g, 38.7 mmol), 80 mL of 1,4-dioxane, and 8 mL of H2O sequentially. Under N2 protection, heat to 80 °C and react for 2 h. TLC (VL) is then performed. 石油醚 :V 乙酸乙酯 =2:1) ​​The reaction was monitored until complete, cooled to room temperature, and half of the solvent was removed by vacuum distillation. The mixture was then extracted with ethyl acetate (40 mL × 2), the ethyl acetate layers were combined and dried over anhydrous sodium sulfate. The solvent was removed by vacuum distillation, and the residue was purified by column chromatography (V... 石油醚 :V 乙酸乙酯 =2:1+TEA) to obtain 2.00g of yellow solid, yield 60.4%.

[0072] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.99 (d, J = 8.6 Hz, 1H), 7.89 (s, 1H), 7.83 (s, 1H), 6.82 (d, J = 8.6 Hz, 1H), 3.87 (s, 3H), 3.77 (s, 3H).

[0073] Synthesis of methyl 4-amino-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (6-1)

[0074] Intermediate 5-1 (1.00 g, 3.90 mmol), 10 mL of anhydrous toluene, and NaNH2 (304 mg, 7.80 mmol) were added sequentially to a 25 mL pear-shaped flask. The reaction system was heated at 100 °C for 2 h under N2 protection. TLC (V1) was used to measure the reaction volume. 二氯甲烷 :V 甲醇 =20:1+TEA) The reaction was monitored to be complete. The reaction solution was cooled to below 5°C, a small amount of ice water was added, the solvent was removed by vacuum evaporation, the residue was slurryed with ice water, filtered, the filter cake was collected, and dried under vacuum to obtain 800 mg of white solid, with a yield of 80.0%.

[0075] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.80 (s, 1H), 8.28 (s, 1H), 8.02 (d, J = 8.8Hz, 1H), 7.60 (d, J = 8.8Hz, 1H), 7.38 (s, 2H), 4.42 (s, 3H), 3.89 (s, 3H).

[0076] Synthesis of 4-amino-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (7-1)

[0077] To a 25 mL pear-shaped flask, intermediate 6-1 (800 mg, 3.10 mmol), 3 mL THF, 3 mL MeOH, and 3 mL H₂O were added sequentially. LiOH (150 mg, 6.24 mmol) was then added under stirring. The mixture was heated to 70 °C and reacted for 2 h. TLC (V1) was then performed. 二氯甲烷 :V 甲醇 =4:1+TEA) The reaction was monitored to be complete. After removing THF and MeOH by vacuum evaporation, 6 mL of H2O was added to the residue, and the pH was adjusted to 3 with 1.5 mol / L HCl. A solid precipitated out. The mixture was filtered, the filter cake was washed with acetone, collected, and dried under vacuum to give 750 mg of white solid, with a yield of 99.0%.

[0078] 1H NMR (300MHz, DMSO-d6) δ (ppm): 8.83 (d, J = 1.8 Hz, 1H), 8.73 (s, 1H), 8.25 (dd, J = 8.6, 1.8 Hz, 1H), 7.90 (d, J = 8.6 Hz, 1H), 4.49 (s, 3H).

[0079] Synthesis of 4-amino-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-acyl chloride hydrochloride (8-1)

[0080] In a 25 mL round-bottom flask, intermediate 7-1 (750 mg, 3.10 mmol) and 5 mL of dichloromethane were added. While stirring, a 4 mol / L dioxane solution (2.33 mL, 9.30 mmol) was added, and the mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure, and the remaining solid was dissolved in 5 mL of dichloromethane. Under ice bath conditions, oxaloyl chloride (1.59 mL, 18.6 mmol) and 2 drops of DMF were added dropwise. After stirring for 5 min, the mixture was moved to room temperature and the reaction was continued for 5 h. TLC (V1) was performed. 二氯甲烷 :V 甲醇 =4:1) Monitor the reaction to ensure it is complete, remove the solvent under reduced pressure, dissolve the residue in 5 mL of dichloromethane, remove the dichloromethane under reduced pressure, repeat the operation once to remove excess oxalyl chloride, and obtain a white solid. No purification is required, and proceed directly to the next step.

[0081] Synthesis of N-((5-bromopyridin-2-yl)methyl)-O-methylhydroxylamine (11-1)

[0082] Add 5-bromopyridine-2-carboxaldehyde 9-1 (794 mg, 4.27 mmol), O-methylhydroxylamine hydrochloride 10-1 (500 mg, 5.99 mmol), and 14 mL of dichloromethane sequentially to a 25 mL round-bottom flask. Add pyridine (378 μL, 4.63 mmol) while stirring at room temperature and continue stirring for 3 h. TLC (V1) is then performed. 石油醚 :V 乙酸乙酯 =10:1) The reaction was monitored to be complete. 14 mL of dichloromethane was added for dilution, followed by washing with 14 mL of 1 mol / L HCl and 14 mL of saturated saline solution, then dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was collected. The solvent was removed by vacuum distillation to obtain an oily liquid, which was directly added to the next reaction without further purification. The obtained oily liquid (360 mg, 1.67 mmol) and 4 mL of glacial acetic acid were added sequentially to a 25 mL round-bottom flask. NaBH3CN (210 mg, 3.35 mmol) was slowly added while stirring. The mixture was stirred at room temperature for 12 h. TLC (V) was performed. 石油醚 :V 乙酸乙酯=1:1) The reaction was monitored until complete. Glacial acetic acid was removed by vacuum distillation. The residue was dissolved in 8 mL of 0.05 mol / L HCl, then washed successively with dichloromethane (8 mL × 3). The pH was adjusted to 9 with saturated Na₂CO₃ solution, and the product was extracted with dichloromethane (8 mL × 3). The organic phases were combined and washed successively with saturated Na₂CO₃ and saturated brine. The product was dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The solvent was removed by vacuum distillation. The crude product was purified by column chromatography (V... 石油醚 :V 乙酸乙酯 =20:1) yielded 160 mg of a colorless oily liquid, with a yield of 17.3%.

[0083] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 8.63 (d, J = 2.4Hz, 1H), 7.77 (dd, J = 8.3, 2.4Hz, 1H), 7.23 (d, J = 8.3Hz, 1H), 4.10 (s, 2H), 3.52 (s, 3H).

[0084] Synthesis of 4-amino-N-((5-bromopyridin-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-1)

[0085] To a 25 mL round-bottom flask, add intermediate 11-1 (85.0 mg, 0.392 mmol) and 1 mL of tetrahydrofuran sequentially. Under ice bath conditions, add DIPEA (287 μL, 1.65 mmol) and intermediate 8-1 dissolved in 2 mL of tetrahydrofuran (122 mg, 0.412 mmol). After 5 min, move the flask to room temperature and react for 5 h. TLC (V 二氯甲烷 :V 甲醇 =20:1+TEA) The reaction was monitored to ensure completeness. 2 mL of tetrahydrofuran was added to the reaction solution, and the solvent was removed by vacuum distillation. The crude product was then subjected to column chromatography (V) 二氯甲烷 :V 氨甲醇(7mmol / l) The solution was purified at a ratio of 150:1 to give 60.0 mg of a white solid, with a yield of 34.7%.

[0086] 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.72(d,J=2.6Hz,1H),8.64(s,1H),8.27(s,1H),8.08(dd,J=8.4,2.6Hz,1H),7.84( d,J=8.7Hz,1H),7.60(d,J=8.7Hz,1H),7.45(d,J=8.4Hz,1H),7.26(s,2H),5.04(s,2H),4.37(s,3H),3.63(s,3H).

[0087] Example 2: 4-Amino-N-methoxy-N-(4-methoxybenzyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-2: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0088] Synthesis of N-(4-methoxybenzyl)-O-methylhydroxylamine (11-2)

[0089] Using 4-methoxybenzaldehyde 9-2 (581 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-1, yielding 135 mg of a colorless oily liquid with a yield of 19.1%.

[0090] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.27 (d, J = 8.5Hz, 2H), 6.88 (d, J = 8.5Hz, 2H), 3.99 (s, 2H), 3.79 (s, 3H), 3.50 (s, 3H).

[0091] Synthesis of 4-amino-N-methoxy-N-(4-methoxybenzyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-2)

[0092] Using intermediates 11-2 (69.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 55.0 mg of white solid with a yield of 35.8%.

[0093] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.56 (s, 1H), 8.26 (s, 1H), 7.79 (d, J = 7.4Hz, 1H), 7.60 (s, 1H), 7.3 3(d,J=6.4Hz,2H),7.25(s,2H),6.95(s,2H),4.89(s,2H),4.34(s,3H),3.74(s,3H),3.60(s,3H).

[0094] Example 3: 4-Amino-N-(2-bromo-4-fluorobenzyl)-N-ethoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-3: R 2 =H,R 4=CH2CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0095] Synthesis of N-(2-bromo-4-fluorobenzyl)-O-ethylhydroxylamine (11-3)

[0096] Using 2-bromo-4-fluorobenzaldehyde 9-3 (866 mg, 4.27 mmol) and O-ethylhydroxylamine hydrochloride 10-2 (584 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 146 mg of colorless oily liquid, with a yield of 13.9%.

[0097] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.38 (dd, J=8.5, 6.0Hz, 1H), 7.30 (dd, J=8.3, 2.6Hz, 1H) ,7.01(td,J=8.3,2.6Hz,1H),4.12(s,2H),3.72(q,J=7.0Hz,2H),1.14(t,J=7.0Hz,3H).

[0098] Synthesis of 4-amino-N-(2-bromo-4-fluorobenzyl)-N-ethoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-3)

[0099] Using intermediates 11-3 (97.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 64.0 mg of white solid with a yield of 34.6%.

[0100] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.61 (s, 1H), 8.28 (s, 1H), 7.84 (dd, J = 8.6, 1.9Hz, 1H), 7.68-7.56 (m, 3H), 7.3 5(dd,J=8.5,2.7Hz,1H),7.29(s,2H),5.03(s,2H),4.36(s,3H),3.81(q,J=7.0Hz,2H),0.91(t,J=7.0Hz,3H).

[0101] Example 4: 4-Amino-N-(4-bromo-2-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-7: R 2 =H,R 4 =CH3,X 1 =X2 =CH, A ring is Synthesis of N-(4-bromo-2-methoxybenzyl)-O-methylhydroxylamine (11-4)

[0102] Using 4-bromo-2-methoxybenzaldehyde 9-4 (918 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 254 mg of a colorless oily liquid, with a yield of 24.0%.

[0103] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.14 (d, J = 7.9Hz, 1H), 7.09-7.04 (m, 1H), 7.00 (d, J = 1.8Hz, 1H), 4.02 (s, 2H), 3.83 (s, 3H), 3.53 (s, 3H).

[0104] Synthesis of 4-amino-N-(4-bromo-2-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-7)

[0105] Using intermediates 11-4 (96.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 76.0 mg of white solid with a yield of 41.0%.

[0106] 1 H NMR(300MHz, DMSO-d6)δ(ppm):8.55(d,J=1.9Hz,1H),8.30(s,1H),7.82(dd,J=8.7,1.9Hz,1H),7.62(d,J=8.6Hz,1H),7.39(s,2H ),7.30(d,J=8.1Hz,1H),7.23(d,J=1.8Hz,1H),7.18(dd,J=8.0,1.9Hz,1H),4.90(s,2H),4.35(s,3H),3.82(s,3H),3.61(s,3H).

[0107] Example 5: 4-Amino-N-(4-fluoro-3-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-9: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0108] Synthesis of N-(4-fluoro-3-methoxybenzyl)-O-methylhydroxylamine (11-5)

[0109] Using 4-fluoro-3-methoxybenzaldehyde 9-5 (658 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 164 mg of colorless oily liquid, with a yield of 20.7%.

[0110] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.07-6.97 (m, 2H), 6.89-6.84 (m, 1H), 4.00 (s, 2H), 3.90 (s, 3H), 3.50 (s, 3H).

[0111] Synthesis of 4-amino-N-(4-fluoro-3-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-9)

[0112] Using intermediates 11-5 (73.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 68.0 mg of white solid with a yield of 42.4%.

[0113] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.59 (s, 1H), 8.45 (s, 1H), 8.24-7.97 (m, 2H), 7.89 (d, J = 9.5Hz, 1H), 7 .71(d,J=8.5Hz,1H),7.19(s,2H),6.99(s,1H),4.96(s,2H),4.38(s,3H),3.84(s,3H),3.62(s,3H).

[0114] Example 6: 4-Amino-N-methoxy-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-10: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of O-methyl-N-(4-(trifluoromethyl)benzyl)hydroxylamine (11-6)

[0115] Using 4-(trifluoromethyl)benzaldehyde 9-6 (743 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 247 mg of colorless oily liquid, with a yield of 28.2%.

[0116] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.60 (d, J = 8.4Hz, 2H), 7.49 (d, J = 8.1Hz, 2H), 4.10 (s, 2H), 3.50 (s, 3H).

[0117] Synthesis of 4-amino-N-methoxy-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-10)

[0118] Using intermediates 11-6 (80.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 64.0 mg of white solid with a yield of 38.0%.

[0119] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.60 (d, J = 2.1Hz, 1H), 8.27 (s, 1H), 7.84 (dd, J = 8.7, 1.9Hz, 1H), 7.77 (d, J = 8 .5Hz,2H),7.65(d,J=8.0Hz,2H),7.62(d,J=8.6Hz,1H),7.28(s,2H),5.10(s,2H),4.35(s,3H),3.66(s,3H).

[0120] Example 7: 4-amino-N-((6-fluoropyridin-3-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-16: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of N-((6-fluoropyridin-3-yl)methyl)-O-methylhydroxylamine (11-7)

[0121] Using 6-fluoropyridine-3-carboxaldehyde 9-7 (534 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 105 mg of colorless oily liquid, with a yield of 15.8%.

[0122] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 8.18 (s, 1H), 7.82 (t, J = 8.1Hz, 1H), 6.91 (dd, J = 7.8, 2.5Hz, 1H), 4.02 (s, 2H), 3.44 (s, 3H).

[0123] Synthesis of 4-amino-N-((6-fluoropyridin-3-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-16)

[0124] Using intermediates 11-7 (61.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 50.0 mg of white solid with a yield of 33.5%.

[0125] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.59 (s, 1H), 8.29 (s, 1H), 8.27 (s, 1H), 8.06 (td, J = 2.2, 8.2Hz, 1H), 7.82 (d, J = 9. 0Hz,1H),7.61(d,J=8.4Hz,1H),7.48(s,2H),7.22(dd,J=2.5,7.0Hz,1H),5.02(s,2H),4.38(s,3H),3.67(s,3H).

[0126] Example 8: 4-Amino-N-(4-bromobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-17: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0127] Synthesis of N-(4-bromobenzyl)-O-methylhydroxylamine (11-8)

[0128] Using 4-bromobenzaldehyde 9-8 (790 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 140 mg of a colorless oily liquid with an overall yield of 15.2%.

[0129] 1H NMR (300MHz, CDCl3-d) δ (ppm): 7.62 (d, J = 8.4Hz, 2H), 7.29 (d, J = 8.4Hz, 2H), 4.83 (s, 2H), 2.24 (s, 3H).

[0130] Synthesis of 4-amino-N-(4-bromobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-17)

[0131] Using intermediates 11-8 (85.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 65.0 mg of white solid with a yield of 37.7%.

[0132] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.57 (s, 1H), 8.26 (s, 1H), 7.80 (d, J = 9.6Hz, 1H), 7.60-7 .57(m,3H),7.37(d,J=7.9Hz,2H),7.23(s,2H),4.96(s,2H),4.35(s,3H),3.63(s,3H).

[0133] Example 9: 4-Amino-N-methoxy-N-((6-methoxypyridin-3-yl)methyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-23: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0134] Synthesis of N-((6-methoxypyridin-3-yl)methyl)-O-methylhydroxylamine (11-9)

[0135] Using 6-methoxypyridine-3-carboxaldehyde 9-9 (585 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 196 mg of a colorless oily liquid, with a yield of 27.3%.

[0136] 1H NMR (400MHz, CDCl3-d) δ (ppm): 8.12 (d, J = 2.5Hz, 1H), 7.61 (dd, J = 8.5, 2.5Hz, 1H), 6.74 (d, J = 8.5Hz, 1H), 3.97 (s, 2H), 3.94 (s, 3H), 3.49 (s, 3H).

[0137] Synthesis of 4-amino-N-methoxy-N-((6-methoxypyridin-3-yl)methyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-23)

[0138] Using intermediates 11-9 (66.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 70.0 mg of white solid with a yield of 45.5%.

[0139] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.56 (s, 1H), 8.27 (s, 1H), 8.20 (s, 1H), 7.82-7.72 (m, 2H), 7.58 (d, J= 6.5Hz,1H),7.32(s,2H),6.84(d,J=8.4Hz,1H),4.92(s,2H),4.36(s,3H),3.84(s,3H),3.65(s,3H).

[0140] Example 10: 4-Amino-N-(benzyloxy)-N-((5-bromopyridin-2-yl)methyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-24: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0141] Synthesis of O-benzyl-N-((5-bromopyridin-2-yl)methyl)hydroxylamine (11-10)

[0142] Using compound 9-1 (794 mg, 4.27 mmol) and O-benzylhydroxylamine 10-3 (737 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 350 mg of colorless oily liquid, with a yield of 30.0%.

[0143] 1H NMR (300MHz, DMSO-d6) δ (ppm): 8.62 (d, J=1.8Hz, 1H), 7.99 (dd, J=8.4, 2.5Hz, 1H), 7.45 (d ,J=8.1Hz,1H),7.33-7.23(m,5H),7.22-7.13(m,1H),4.59(s,2H),4.03(d,J=4.9Hz,2H).

[0144] Synthesis of 4-amino-N-(benzyloxy)-N-((5-bromopyridin-2-yl)methyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-24)

[0145] Using intermediates 11-10 (115 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 75.0 mg of white solid with a yield of 37.0%.

[0146] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.71 (d, J = 2.4Hz, 1H), 8.53 (d, J = 1.9Hz, 1H), 8.35 (s, 1H), 8.07 (dd, J = 8.3, 2.5Hz, 1H), 7.83 (dd, J = 8.7, 1.9Hz,1H),7.63(d,J=8.6Hz,2H),7.46(d,J=8.4Hz,1H),7.30-7.11(m,4H),7.08-7.02(m,2H),5.08(s,2H),4.82(s,2H),4.21(s,3H).

[0147] Example 11: 4-Amino-N-methoxy-1-methyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-25: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0148] Synthesis of O-methyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)hydroxylamine (11-11)

[0149] Using 6-(trifluoromethyl)pyridine-3-carboxaldehyde 9-10 (747 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 128 mg of colorless oily liquid, with a yield of 14.5%.

[0150] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 8.71 (s, 1H), 7.90 (d, J = 8.0Hz, 1H), 7.67 (d, J = 8.0Hz, 1H), 4.12 (s, 2H), 3.45 (s, 3H).

[0151] Synthesis of 4-amino-N-methoxy-1-methyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-25)

[0152] Using intermediates 11-11 (89.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 45.0 mg of white solid with a yield of 26.7%.

[0153] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.83 (s, 1H), 8.61 (d, J = 2.0Hz, 1H), 8.28 (s, 1H), 8.12 (d, J = 7.8Hz, 1H), 7.95 (d, J = 7 .8Hz,1H),7.84(dd,J=8.7,2.0Hz,1H),7.60(d,J=8.7Hz,1H),7.38(s,1H),5.15(s,2H),4.37(s,3H),3.70(s,3H).

[0154] Example 12: 4-Amino-N-((5-bromopyridin-2-yl)methyl)-N-(tert-butoxy)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-31: R 2 =H,R 4 =C(CH3)3,X 1 =X 2 =CH, A ring is Synthesis of )

[0155] Synthesis of N-((5-bromopyridin-2-yl)methyl)-O-(tert-butyl)hydroxylamine (11-12)

[0156] Using compound 9-1 (794 mg, 4.27 mmol) and O-tert-butylhydroxylamine 10-4 (752 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-1, yielding 330 mg of a colorless oily liquid with a yield of 29.8%.

[0157] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 8.63 (d, J = 3.0Hz, 1H), 7.77 (dd, J = 8.2, 2.3Hz, 1H), 7.23 (d, J = 8.1Hz, 1H), 4.05 (s, 2H), 1.15 (s, 9H).

[0158] Synthesis of 4-amino-N-((5-bromopyridin-2-yl)methyl)-N-(tert-butoxy)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-31)

[0159] Using intermediates 11-12 (102 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 64.0 mg of white solid with a yield of 33.8%.

[0160] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.69 (d, J=2.5Hz, 1H), 8.60 (s, 1H), 8.32 (s, 1H), 8.05 (dd, J=8.4, 2.5Hz, 1H), 7.81 ( d,J=8.6Hz,1H),7.61(d,J=8.6Hz,1H),7.46(s,2H),7.40(d,J=8.4Hz,1H),5.02(s,2H),4.37(s,3H),1.07(s,9H).

[0161] Example 13: N-(allyloxy)-4-amino-N-((5-bromopyridin-2-yl)methyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-32: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0162] Synthesis of O-allyl-N-((5-bromopyridin-2-yl)methyl)hydroxylamine (11-13)

[0163] Using compound 9-1 (794 mg, 4.27 mmol) and O-allyl hydroxylamine hydrochloride 10-5 (656 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 265 mg of colorless oily liquid, with a yield of 25.5%.

[0164] 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.61(d,J=2.1Hz,1H),8.01(dd,J=8.3,2.4Hz,1H),7.45(d,J=8.4Hz,1H),7.1 2(t,J=6.0Hz,1H),5.92-5.76(m,1H),5.22-5.05(m,2H),4.06(dt,J=5.6,1.4Hz,2H),4.00(d,J=6.0Hz,2H).

[0165] Synthesis of N-(allyloxy)-4-amino-N-((5-bromopyridin-2-yl)methyl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-32)

[0166] Using intermediates 11-13 (95.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 45.0 mg of white solid with a yield of 24.6%.

[0167] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.72 (s, 1H), 8.63 (s, 1H), 8.26 (s, 1H), 8.08 (d, J = 10.9Hz, 1H), 7.83 (d, J = 9.4Hz, 1H), 7. 59(d,J=8.8Hz,1H),7.46(d,J=8.4Hz,1H),7.26(s,2H),5.78-5.62(m,1H),5.24-5.11(m,2H),5.05(s,2H),4.35(s,5H).

[0168] Example 14: (E)-4-amino-N-((5-bromopyridin-2-yl)methyl)-N-((3-chloroallyl)oxy)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-36: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0169] Synthesis of (E)-N-((5-bromopyridin-2-yl)methyl)-O-(3-chloroallyl)hydroxylamine (11-14)

[0170] Using compound 9-1 (794 mg, 4.27 mmol) and trans-3-chloro-2-propenylhydroxylamine 10-6 (644 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 237 mg of colorless oily liquid, with a yield of 20.0%.

[0171] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.61 (s, 1H), 8.01 (d, J = 9.3Hz, 1H), 7.44 (d, J = 8.3Hz, 1H), 7.20 (t, J = 6.2H z,1H),6.44(d,J=13.2Hz,1H),5.98(dt,J=12.9,6.5Hz,1H),4.05(d,J=6.5Hz,2H),3.99(d,J=6.3Hz,2H).

[0172] Synthesis of (E)-4-amino-N-((5-bromopyridin-2-yl)methyl)-N-((3-chloroallyl)oxy)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-36)

[0173] Using intermediates 11-14 (109 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 63.0 mg of white solid with a yield of 32.0%.

[0174] 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.72(d,J=1.9Hz,1H),8.58(d,J=1.9Hz,1H),8.26(s,1H),8.08(dd,J=8.3,2.5Hz,1H),7.79(dd,J=8.7,1.9H z,1H),7.59(d,J=8.6Hz,1H),7.44(d,J=8.5Hz,1H),7.25(s,2H),6.53(d,J=14.4Hz,1H),5.79-5.75(m,1H),5.01(s,2H),4.39-4.32(m,5H).

[0175] Example 15: 4-Amino-N-((5-bromopyridin-2-yl)methyl)-N-((4-fluorobenzyl)oxy)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-38: R 2=H, X 1 =X 2 =CH, A ring is Synthesis of )

[0176] Synthesis of N-((5-bromopyridin-2-yl)methyl)-O-(4-fluorobenzyl)hydroxylamine (11-15)

[0177] Using compound 9-1 (794 mg, 4.27 mmol) and o-(4-fluorobenzyl)hydroxylamine hydrochloride 10-7 (1.06 g, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 312 mg of colorless oily liquid, with a yield of 23.5%.

[0178] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 8.40 (d, J = 3.1Hz, 1H), 7.88 (dd, J = 8.2, 2.3Hz, 2H), 7.17-7.13 (m, 2H), 7.07-6.99 (m, 2H), 4.98 (s, 2H), 4.73 (s, 2H).

[0179] Synthesis of 4-amino-N-((5-bromopyridin-2-yl)methyl)-N-((4-fluorobenzyl)oxy)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-38)

[0180] Using intermediates 11-15 (122 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 85.0 mg of white solid with a yield of 40.5%.

[0181] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.72 (d, J = 2.4Hz, 1H), 8.50 (d, J = 1.9Hz, 1H), 8.27 (s, 1H), 8.08 (dd, J = 8.3, 2.5Hz, 1H), 7.79 (dd, J = 8.7, 2.0Hz, 1H), 7 .58(d,J=8.7Hz,1H),7.46(d,J=8.3Hz,1H),7.25(s,2H),7.13(dd,J=8.8, 5.6Hz,2H),7.00(t,J=8.9Hz,2H),5.08(s,2H),4.83(s,2H),4.25(s,3H).

[0182] Example 16: 4-Amino-N-methoxy-1-methyl-N-(4-morpholinyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-39: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0183] Synthesis of O-methyl-N-(4-morpholinobenzyl)hydroxylamine (11-16)

[0184] Using 4-morpholinobenzaldehyde 9-11 (816 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 302 mg of colorless oily liquid, with a yield of 31.8%.

[0185] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.30 (s, 1H), 7.27 (s, 1H), 6.94-6.89 (m, 2H), 4.00 (s, 2H), 3.90-3.86 (m, 4H), 3.54 (s, 3H), 3.20-3.14 (m, 4H).

[0186] Synthesis of 4-amino-N-methoxy-1-methyl-N-(4-morpholinyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-39)

[0187] Using intermediates 11-16 (87.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 70.0 mg of white solid with a yield of 40.0%.

[0188] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.54 (d, J = 2.1Hz, 1H), 8.28 (s, 1H), 7.79 (dd, J = 8.6, 1.9Hz, 1H), 7.60 (d, J = 8.7Hz, 1H), 7.36 (s, 1H),7.27(d,J=8.7Hz,2H),6.94(d,J=8.8Hz,2H),4.86(s,2H),4.34(s,3H),3.76-3.71(m,4H),3.60(s,3H),3.12-3.07(m,4H).

[0189] Example 17: 4-Amino-N-(4-fluorobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-40: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0190] Synthesis of N-(4-fluorobenzyl)-O-methylhydroxylamine (11-17)

[0191] Using 4-fluorobenzaldehyde 9-12 (530 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 320 mg of colorless oily liquid, with a yield of 48.0%.

[0192] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.36-7.29 (m, 2H), 7.07-6.98 (m, 2H), 4.01 (s, 2H), 3.49 (s, 3H).

[0193] Synthesis of 4-amino-N-(4-fluorobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-40)

[0194] Using intermediates 11-17 (61.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 65.0 mg of white solid with a yield of 44.0%.

[0195] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.58 (d, J=2.0Hz, 1H), 8.30 (s, 1H), 7.82 (dd, J=8.6, 2.0Hz, 1H), 7.61 (d, J= 8.6Hz,1H),7.50-7.44(m,2H),7.43(s,2H),7.21(t,J=8.9Hz,2H),4.97(s,2H),4.36(s,3H),3.62(s,3H).

[0196] Example 18: 4-Amino-N-(4-(dimethylamino)benzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-41: R 2 =H,R 4 =CH3,X1 =X 2 =CH, A ring is Synthesis of )

[0197] Synthesis of 4-((methoxyamino)methyl)-N,N-dimethylaniline (11-18)

[0198] Using 4-(dimethylamino)benzaldehyde 9-13 (637 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 342 mg of colorless oily liquid, with a yield of 44.5%.

[0199] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 7.22 (d, J = 8.8 Hz, 2H), 6.72 (d, J = 8.8 Hz, 2H), 3.97 (s, 2H), 3.53 (s, 3H), 2.94 (s, 6H).

[0200] Synthesis of 4-amino-N-(4-(dimethylamino)benzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-41)

[0201] Using intermediates 11-18 (71.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 68.0 mg of white solid with a yield of 42.9%.

[0202] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.52 (s, 1H), 8.26 (s, 1H), 7.77 (d, J = 8.6Hz, 1H), 7.59 (d, J = 8.8Hz, 1H),7.33-7.17(m,4H),6.71(d,J=8.6Hz,2H),4.81(s,2H),4.33(s,3H),3.57(s,3H),2.87(s,6H).

[0203] Example 19: 4-Amino-N-(4-chloro-3-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-42: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of N-(4-chloro-3-methoxybenzyl)-O-methylhydroxylamine (11-19)

[0204] Using 4-chloro-3-methoxybenzaldehyde 9-14 (728 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 410 mg of a colorless oily liquid with a yield of 47.6%.

[0205] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.31 (d, J = 8.0Hz, 1H), 6.96 (d, J = 1.9Hz, 1H), 6.89 (dd, J = 8.1, 1.9Hz, 1H), 4.01 (s, 2H), 3.91 (s, 3H), 3.50 (s, 3H).

[0206] Synthesis of 4-amino-N-(4-chloro-3-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-42)

[0207] Using intermediates 11-19 (79.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 62.0 mg of white solid with a yield of 37.0%.

[0208] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.57 (s, 1H), 8.35 (s, 1H), 7.83 (d, J = 8.3Hz, 1H), 7.62 (d, J = 8.7Hz, 1H), 7. 57-7.36(m,3H),7.16(s,1H),7.01(d,J=7.8Hz,1H),4.98(s,2H),4.35(s,3H),3.85(s,3H),3.63(s,3H).

[0209] Example 20: 4-Amino-N-(4-cyanobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-48: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0210] Synthesis of 4-((methoxyamino)methyl)benzyl nitrile (11-20)

[0211] Using 4-cyanobenzaldehyde 9-15 (560 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 255 mg of colorless oily liquid, with a yield of 36.8%.

[0212] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 6.57 (d, J = 8.5 Hz, 2H), 6.35 (d, J = 8.4 Hz, 2H), 2.79 (s, 2H), 2.14 (s, 3H).

[0213] Synthesis of 4-amino-N-(4-cyanobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-48)

[0214] Using intermediates 11-20 (64.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 45.0 mg of white solid with a yield of 29.7%.

[0215] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.60 (d, J = 2.0Hz, 1H), 8.35 (s, 1H), 7.86-7.83 ( m,3H),7.62-7.59(m,3H),7.47(s,2H),5.09(s,2H),4.36(s,3H),3.65(s,3H).

[0216] Example 21: 4-Amino-N-(4-(difluoromethoxy)-3-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-54: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0217] Synthesis of N-(4-(difluoromethoxy)-3-methoxybenzyl)-O-methylhydroxylamine (11-21)

[0218] Using 4-(difluoromethoxy)-3-methoxybenzaldehyde 9-16 (863 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 230 mg of colorless oily liquid, with a yield of 23.1%.

[0219] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.12 (d, J = 8.1Hz, 1H), 7.00 (d, J = 2.1Hz, 1H), 6.91 ( dd,J=8.1,2.1Hz,1H),6.53(t,J=75Hz,1H),4.02(s,2H),3.89(s,3H),3.52(s,3H).

[0220] Synthesis of 4-amino-N-(4-(difluoromethoxy)-3-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-54)

[0221] Using intermediates 11-21 (91.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 52.0 mg of white solid with a yield of 29.0%.

[0222] 1 H NMR(400MHz, DMSO-d6)δ(ppm):8.57(d,J=2.0Hz,1H),8.31(s,1H),7.81(dd,J=8.6,2.0Hz,1H),7.59(d,J=8.6Hz,1H),7.33(s,2H),7 .20-7.15(m,2H),7.06(s,1H),7.06(t,J=76Hz,1H),7.00(dd,J=8.3,2.0Hz,1H),4.97(s,2H),4.33(s,3H),3.82(s,3H),3.63(s,3H).

[0223] Example 22: 4-amino-N-((5-bromofuran-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-55: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0224] Synthesis of N-((5-bromofuran-2-yl)methyl)-O-methylhydroxylamine (11-22)

[0225] Using 5-bromofuran-2-carboxaldehyde 9-17 (747 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 324 mg of a colorless oily liquid with a yield of 36.8%.

[0226] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 6.26-6.24 (m, 2H), 4.01 (s, 2H), 3.52 (s, 3H).

[0227] Synthesis of 4-amino-N-((5-bromofuran-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-55)

[0228] Using intermediates 11-22 (81.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 61.0 mg of white solid with a yield of 36.2%.

[0229] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.56 (d, J=2.0Hz, 1H), 8.31 (s, 1H), 7.78 (dd, J=8.7, 2.0Hz, 1H) ,7.59(d,J=8.6Hz,1H),7.32(s,2H),6.58-6.52(m,2H),4.94(s,2H),4.39(s,3H),3.61(s,3H).

[0230] Example 23: 4-amino-N-((5-chloropyridin-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-56: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of N-((5-chloropyridin-2-yl)methyl)-O-methylhydroxylamine (11-23)

[0231] Using 5-chloropyridine-2-carboxaldehyde 9-18 (606 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 210 mg of a colorless oily liquid, with a yield of 28.5%.

[0232] 1H NMR (300MHz, CDCl3-d) δ (ppm): 8.53 (d, J = 2.5Hz, 1H), 7.63 (dd, J = 8.3, 2.5Hz, 1H), 7.28 (d, J = 8.9Hz, 1H), 4.13 (s, 2H), 3.52 (s, 3H).

[0233] Synthesis of 4-amino-N-((5-chloropyridin-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-56)

[0234] Using intermediates 11-23 (68.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 51.0 mg of white solid with a yield of 32.8%.

[0235] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.63 (d, J = 2.1Hz, 2H), 8.30 (s, 1H), 7.96 (dd, J = 8.4, 2.5Hz, 1H), 7.83 (dd, J = 8.7 ,2.0Hz,1H),7.58(d,J=8.7Hz,1H),7.50(d,J=8.4Hz,1H),7.29(s,2H),5.06(s,2H),4.36(s,3H),3.62(s,3H).

[0236] Example 24: 4-Amino-N-(4-cyano-2-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-60: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0237] Synthesis of 3-methoxy-4-((methoxyamino)methyl)benzyl nitrile (11-24)

[0238] Using 4-cyano-3-methoxybenzaldehyde 9-19 (688 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 230 mg of colorless oily liquid, with a yield of 28.0%.

[0239] 1H NMR (300MHz, CDCl3-d) δ (ppm): 7.39 (d, J = 7.7Hz, 1H), 7.24 (d, J = 1.5Hz, 1H), 7.09 (d, J = 1.5Hz, 1H), 4.10 (s, 2H), 3.88 (s, 3H), 3.53 (s, 3H).

[0240] Synthesis of 4-amino-N-(4-cyano-2-methoxybenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-60)

[0241] Using intermediates 11-24 (75.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 60.0 mg of white solid with a yield of 36.8%.

[0242] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.58 (s, 1H), 8.28 (s, 1H), 7.83 (d, J = 9.5Hz, 1H), 7.60 (d, J = 7. 9Hz,1H),7.52-7.45(m,3H),7.30(s,2H),4.99(s,2H),4.35(s,3H),3.87(s,3H),3.64(s,3H).

[0243] Example 25: 4-amino-N-((5-chloropyrazin-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-63: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0244] Synthesis of N-((5-chloropyrazin-2-yl)methyl)-O-methylhydroxylamine (11-25)

[0245] Using 5-chloropyrazine-2-carboxaldehyde 9-20 (608 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 180 mg of a colorless oily liquid, with a yield of 24.3%.

[0246] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 8.56 (s, 1H), 8.40 (s, 1H), 4.17 (s, 2H), 3.51 (s, 3H).

[0247] Synthesis of 4-amino-N-((5-chloropyrazin-2-yl)methyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-63)

[0248] Using intermediates 11-25 (56.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 57.0 mg of white solid with a yield of 36.6%.

[0249] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.83 (d, J = 1.5 Hz, 1H), 8.63 (dd, J = 7.8, 1.8 Hz, 2H), 8.32 (s, 1H), 7.8 3(dd,J=8.6,2.0Hz,1H),7.59(d,J=8.8Hz,1H),7.34(s,2H),5.15(s,2H),4.38(s,3H),3.66(s,3H).

[0250] Example 26: 4-Amino-N-(4-cyano-2-fluorobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-64: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0251] Synthesis of 3-fluoro-4-((methoxyamino)methyl)benzylnitrile (11-26)

[0252] Using 4-cyano-2-fluorobenzaldehyde 9-21 (636 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 230 mg of colorless oily liquid, with a yield of 30.0%.

[0253] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.56 (t, J = 7.6 Hz, 1H), 7.45 (d, J = 7.8 Hz, 1H), 7.35 (d, J = 9.2 Hz, 1H), 4.14 (s, 2H), 3.49 (s, 3H).

[0254] Synthesis of 4-amino-N-(4-cyano-2-fluorobenzyl)-N-methoxy-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-64)

[0255] Using intermediates 11-26 (71.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 60.0 mg of white solid with a yield of 37.8%.

[0256] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.60 (s, 1H), 8.31 (s, 1H), 7.91 (d, J = 10.0Hz, 1H), 7.82 (d, J = 9.1Hz, 1H),7.75-7.69(m,2H),7.59(d,J=8.8Hz,1H),7.31(s,2H),5.12(s,2H),4.37(s,3H),3.65(s,3H).

[0257] Example 26: (E)-4-amino-N-((3-chloroallyl)oxy)-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-65: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0258] Synthesis of (E)-O-(3-chloroallyl)-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)hydroxylamine (11-27)

[0259] Using 4-(trifluoromethyl)benzaldehyde 9-6 (743 mg, 4.27 mmol) and trans-3-chloro-2-propenylhydroxylamine 10-6 (644 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 182.0 mg of a colorless oily liquid with a yield of 16.1%.

[0260] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.75 (d, J = 8.0Hz, 2H), 7.62 (d, J = 7.5Hz, 2H), 6.35 (d, J = 13.3Hz, 1H), 6.15-6.03 (m, 1H), 4.24 (s, 2H).

[0261] Synthesis of (E)-4-amino-N-((3-chloroallyl)oxy)-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-65)

[0262] Using intermediates 11-27 (182.0 mg, 0.687 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 52.0 mg of white solid with a yield of 26.0%.

[0263] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.54 (s, 1H), 8.53 (s, 1H), 7.87 (d, J = 8.6Hz, 1H), 7.77 (d, J = 8.1Hz, 2H), 7.72 (d, J = 8.6 Hz,1H),7.64(d,J=8.0Hz,2H),6.54(d,J=13.2Hz,1H),5.80(m,1H),5.10(s,2H),4.39(d,J=7.3Hz,2H),4.37(s,3H).

[0264] Example 27: 4-Amino-N-(4-chloro-3-methoxybenzyl)-N-methoxy-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-66: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0265] Synthesis of methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)benzoate (2)

[0266] To a 250 mL round-bottom flask, methyl 4-amino-3-bromobenzoate 1 (4.00 g, 17.4 mmol), pinacol diborate (8.83 g, 34.8 mmol), and 60 mL of 1,4-dioxane were added sequentially. Potassium acetate (5.12 g, 52.2 mmol) and Pd(dppf)Cl2 (1.42 g, 1.74 mmol) were then added under stirring at room temperature. The reaction system was refluxed at 100 °C for 18 h under N2 protection. TLC (VL) was performed. 石油醚 :V 乙酸乙酯The reaction was monitored to ensure complete reaction (1:1 ratio), cooled to room temperature, and filtered. The filter cake was washed with dichloromethane, and the filtrate was concentrated to obtain a semi-solid residue. This residue was then dissolved in a small amount of dichloromethane, filtered, and the filter cake was collected. The filtrate was concentrated, and the operation was repeated once. The two filter cakes were combined and dried to obtain 3.00 g of a grayish-white solid, with a yield of 62.2%.

[0267] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 8.30 (d, J = 2.2Hz, 1H), 7.87 (dd, J = 8.6, 2.2Hz, 1H), 6.55 (d, J = 8.6Hz, 1H), 3.84 (s, 3H), 1.34 (s, 12H).

[0268] Synthesis of 5-iodo-1,3-dimethyl-1H-pyrazole-4-onitrile (4-2)

[0269] Add 5-amino-1,3-dimethyl-1H-pyrazole-4-onitrile 3-2 (5.00 g, 36.7 mmol) and 90 mL of CH2I2 sequentially to a 250 mL three-necked flask. Heat to 80 °C, then add amyl nitrite (11.0 mL, 81.9 mmol) dropwise. After the addition is complete, continue heating at 80 °C for 1 h. TLC (V 石油醚 :V 乙酸乙酯 =10:1) Monitor the reaction to ensure it is complete, cool to room temperature, extract the reaction solution with concentrated HCl (30 mL × 2), combine the concentrated HCl layers, adjust the pH of the aqueous layer to 1 with saturated NaOH aqueous solution, then extract with ethyl acetate (30 mL × 2), combine the ethyl acetate layers, remove the solvent under reduced pressure, and precipitate the residue by column chromatography (V... 石油醚 :V 乙酸乙酯 Purification with a ratio of 20:1 yielded 3.70 g of a white solid, with a yield of 38.9%.

[0270] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 3.93 (s, 3H), 2.40 (s, 3H).

[0271] Synthesis of methyl 4-amino-3-(4-cyano-1,3-dimethyl-1H-pyrazol-5-yl)benzoate (5-2)

[0272] To a 250 mL pear-shaped flask, add intermediate 2 (4.82 g, 17.5 mmol), intermediate 4-2 (3.70 g, 15.9 mmol), Pd(PPh3)4 (892.8 mg, 0.792 mmol), cesium carbonate (15.5 g, 47.6 mmol), 80 mL of 1,4-dioxane, and 8 mL of H2O sequentially. Under N2 protection, heat to 80 °C and react for 2 h. TLC (VL) is then performed. 石油醚 :V乙酸乙酯 =2:1) ​​The reaction was monitored until complete, cooled to room temperature, and half of the solvent was removed by vacuum distillation. The mixture was then extracted with ethyl acetate (40 mL × 2), the ethyl acetate layers were combined and dried over anhydrous sodium sulfate. The solvent was removed by vacuum distillation, and the residue was purified by column chromatography (V... 石油醚 :V 乙酸乙酯 =2:1+TEA) yielded 2.60g of yellow solid, with a yield of 78.5%.

[0273] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 7.81 (d, J = 10.5 Hz, 1H), 7.62 (s, 1H), 6.85 (d, J = 8.7 Hz, 1H), 6.19 (s, 2H), 3.78 (s, 3H), 3.35 (s, 3H), 2.33 (s, 3H).

[0274] Synthesis of methyl 4-amino-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (6-2)

[0275] Intermediate 5-2 (1.00 g, 3.90 mmol), 10 mL of anhydrous toluene, and NaNH2 (304 mg, 7.80 mmol) were added sequentially to a 25 mL pear-shaped flask. The reaction system was heated at 100 °C for 2 h under N2 protection. TLC (V1) was performed. 二氯甲烷 :V 甲醇 =20:1+TEA) The reaction was monitored to be complete. The reaction solution was cooled to below 5°C, a small amount of ice water was added, the solvent was removed by vacuum evaporation, the residue was slurryed with ice water, filtered, the filter cake was collected, and dried under vacuum to obtain 830 mg of white solid, with a yield of 83.0%.

[0276] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.80 (s, 1H), 8.28 (s, 1H), 8.02 (d, J = 8.8Hz, 1H), 7.38 (s, 2H), 4.42 (s, 3H), 3.89 (s, 3H), 2.06 (s, 3H).

[0277] Synthesis of 4-amino-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (7-2)

[0278] To a 25 mL pear-shaped flask, intermediate 6-2 (830 mg, 3.22 mmol), 3 mL THF, 3 mL MeOH, and 3 mL H₂O were added sequentially. LiOH (155 mg, 6.48 mmol) was then added under stirring. The mixture was heated to 70 °C and reacted for 2 h. TLC (V1) was then performed. 二氯甲烷 :V 甲醇=4:1+TEA) The reaction was monitored to be complete. After removing THF and MeOH by vacuum distillation, 6 mL of H2O was added to the residue, and the pH was adjusted to 3 with 1.5 mol / L HCl. A solid precipitated out. The mixture was filtered, the filter cake was washed with acetone, collected, and dried under vacuum to give 739 mg of white solid, with a yield of 97.5%.

[0279] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.80 (s, 1H), 8.25 (d, J = 10.4Hz, 1H), 7.82 (d, J = 8.6Hz, 1H), 4.41 (s, 3H), 2.66 (s, 3H).

[0280] Synthesis of 4-amino-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-acyl chloride hydrochloride (8-2)

[0281] In a 25 mL round-bottom flask, intermediate 7-2 (739 mg, 3.05 mmol) and 5 mL of dichloromethane were added. While stirring, a 4 mol / L dioxane solution (2.33 mL, 9.30 mmol) was added, and the mixture was stirred at room temperature for 2 h. The solvent was removed by vacuum distillation, and the remaining solid was dissolved in 5 mL of dichloromethane. Under ice bath conditions, oxaloyl chloride (1.59 mL, 18.6 mmol) and 2 drops of DMF were added dropwise. After stirring for 5 min, the mixture was moved to room temperature and the reaction was continued for 5 h. TLC (V1) was performed. 二氯甲烷 :V 甲醇 =4:1) Monitor the reaction to ensure it is complete, remove the solvent under reduced pressure, dissolve the residue in 5 mL of dichloromethane, remove the dichloromethane under reduced pressure, repeat the operation once to remove excess oxalyl chloride, and obtain a white solid. No purification is required, and proceed directly to the next step.

[0282] Synthesis of N-(4-chloro-3-methoxybenzyl)-O-methylhydroxylamine (11-19)

[0283] Using 4-chloro-3-methoxybenzaldehyde 9-14 (728 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 410 mg of a colorless oily liquid with a yield of 47.6%.

[0284] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.31 (d, J = 8.0Hz, 1H), 6.96 (d, J = 1.9Hz, 1H), 6.89 (dd, J = 8.1, 1.9Hz, 1H), 4.01 (s, 2H), 3.91 (s, 3H), 3.50 (s, 3H).

[0285] Synthesis of 4-amino-N-(4-chloro-3-methoxybenzyl)-N-methoxy-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-66)

[0286] Using intermediates 11-19 (80.0 mg, 0.392 mmol) and 8-2 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that in I-1, yielding 74.0 mg of white solid with a yield of 44.0%.

[0287] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.82 (s, 1H), 8.58 (d, J = 17.4Hz, 1H), 7.83 (d, J = 7.3Hz, 1H), 7.43 (d, J = 8.1Hz ,1H),7.16(s,1H),7.02(d,J=8.1Hz,1H),4.99(s,2H),4.29(s,3H),3.21(s,3H),3.15(s,3H),2.67(s,3H).

[0288] Example 28: 4-Amino-N-(4-cyanobenzyl)-N-methoxy-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-67: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0289] Synthesis of 4-((methoxyamino)methyl)benzyl nitrile (11-20)

[0290] Using 4-cyanobenzaldehyde 9-15 (560 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 255 mg of colorless oily liquid, with a yield of 36.8%.

[0291] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 6.57 (d, J = 8.5 Hz, 2H), 6.35 (d, J = 8.4 Hz, 2H), 2.79 (s, 2H), 2.14 (s, 3H).

[0292] Synthesis of 4-amino-N-(4-cyanobenzyl)-N-methoxy-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-67)

[0293] Using intermediates 11-20 (64.0 mg, 0.392 mmol) and 8-2 (122 mg, 0.406 mmol) as raw materials, the preparation method was the same as that in I-1, yielding 59.0 mg of white solid with a yield of 38.9%.

[0294] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.58 (s, 1H), 7.87 (d, J = 8.2Hz, 2H), 7.83 (dd, J = 8.7, 1.9Hz, 1H), 7. 61(d,J=6.8Hz,2H),7.58(s,1H),6.76(s,2H),5.09(s,2H),4.27(s,3H),3.66(s,3H),2.61(s,3H).

[0295] Example 29: 4-Amino-N-ethoxy-1-ethyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-68: R 2 =H,R 4 =CH2CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0296] Synthesis of methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)benzoate (2)

[0297] To a 250 mL round-bottom flask, methyl 4-amino-3-bromobenzoate 1 (4.00 g, 17.4 mmol), pinacol diborate (8.83 g, 34.8 mmol), and 60 mL of 1,4-dioxane were added sequentially. Potassium acetate (5.12 g, 52.2 mmol) and Pd(dppf)Cl2 (1.42 g, 1.74 mmol) were then added under stirring at room temperature. The reaction system was refluxed at 100 °C for 18 h under N2 protection. TLC (VL) was performed. 石油醚 :V 乙酸乙酯 The reaction was monitored to ensure complete reaction (1:1 ratio), cooled to room temperature, and filtered. The filter cake was washed with dichloromethane, and the filtrate was concentrated to obtain a semi-solid residue. This residue was then dissolved in a small amount of dichloromethane, filtered, and the filter cake was collected. The filtrate was concentrated, and the operation was repeated once. The two filter cakes were combined and dried to obtain 3.00 g of a grayish-white solid, with a yield of 62.2%.

[0298] 1H NMR (300MHz, CDCl3-d) δ (ppm): 8.30 (d, J = 2.2Hz, 1H), 7.87 (dd, J = 8.6, 2.2Hz, 1H), 6.55 (d, J = 8.6Hz, 1H), 3.84 (s, 3H), 1.34 (s, 12H).

[0299] Synthesis of 5-iodo-1-ethyl-1H-pyrazole-4-onitrile (4-3)

[0300] Add 5-amino-1-ethyl-1H-pyrazole-4-onitrile 3-3 (5.00 g, 36.7 mmol) and 90 mL of CH2I2 sequentially to a 250 mL three-necked flask, heat to 80 °C, and add amyl nitrite (11.0 mL, 81.9 mmol) dropwise. After the addition is complete, continue heating at 80 °C for 1 h. TLC (V 石油醚 :V 乙酸乙酯 =10:1) Monitor the reaction to ensure it is complete, cool to room temperature, extract the reaction solution with concentrated HCl (30 mL × 2), combine the concentrated HCl layers, adjust the pH of the aqueous layer to 1 with saturated NaOH aqueous solution, then extract with ethyl acetate (30 mL × 2), combine the ethyl acetate layers, remove the solvent under reduced pressure, and precipitate the residue by column chromatography (V... 石油醚 :V 乙酸乙酯 Purification with a ratio of 20:1 yielded 3.90 g of a white solid, with a yield of 41.0%.

[0301] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.88 (s, 1H), 4.32 (q, J = 6.6, 5.9Hz, 2H), 1.50 (t, J = 7.3Hz, 3H).

[0302] Synthesis of methyl 4-amino-3-(4-cyano-1-ethyl-1H-pyrazole-5-yl)benzoate (5-3)

[0303] To a 250 mL pear-shaped flask, add intermediate 2 (5.10 mg, 18.5 mmol), intermediate 4-3 (3.90 g, 14.4 mmol), Pd(PPh3)4 (967.2 mg, 0.837 mmol), cesium carbonate (16.4 g, 50.3 mmol), 80 mL of 1,4-dioxane, and 8 mL of H2O sequentially. Under N2 protection, heat to 80 °C and react for 2 h. TLC (VL) is then performed. 石油醚 :V 乙酸乙酯 =2:1) ​​The reaction was monitored until complete, cooled to room temperature, and half of the solvent was removed by vacuum distillation. The mixture was then extracted with ethyl acetate (40 mL × 2), the ethyl acetate layers were combined and dried over anhydrous sodium sulfate. The solvent was removed by vacuum distillation, and the residue was purified by column chromatography (V... 石油醚 :V 乙酸乙酯=2:1+TEA) to obtain 3.00g of yellow solid, yield 80.6%.

[0304] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.28 (s, 1H), 7.90 (d, J = 6.6Hz, 1H), 7.70 (s, 1H), 6.94 (d,J=8.7Hz,1H),6.24(s,2H),4.12-3.95(m,2H),3.86(s,3H),1.36(t,J=7.2Hz,3H).

[0305] Synthesis of methyl 4-amino-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (6-3)

[0306] Intermediate 5-3 (1.00 g, 3.70 mmol), 10 mL of anhydrous toluene, and NaNH2 (304 mg, 7.80 mmol) were added sequentially to a 25 mL pear-shaped flask. The reaction system was heated at 100 °C for 2 h under N2 protection. TLC (V1) was used to measure the reaction volume. 二氯甲烷 :V 甲醇 =20:1+TEA) The reaction was monitored to be complete. The reaction solution was cooled to below 5°C, a small amount of ice water was added, the solvent was removed by vacuum evaporation, the residue was slurryed with ice water, filtered, the filter cake was collected, and dried under vacuum to obtain 890 mg of white solid, with a yield of 89.0%.

[0307] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.80 (s, 1H), 8.28 (s, 1H), 8.02 (d, J = 8.8Hz, 1H), 7.38 (s, 2H), 4.42 (s, 3H), 3.79 (s, 2H), 1.97 (s, 3H).

[0308] Synthesis of 4-amino-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (7-3)

[0309] To a 25 mL pear-shaped flask, intermediate 6-3 (890 mg, 3.47 mmol), 3 mL THF, 3 mL MeOH, and 3 mL H₂O were added sequentially. LiOH (150 mg, 6.24 mmol) was then added under stirring. The mixture was heated to 70 °C and reacted for 2 h. TLC (V1) was then performed. 二氯甲烷 :V 甲醇 =4:1+TEA) The reaction was monitored to be complete. After removing THF and MeOH by vacuum distillation, 6 mL of H2O was added to the residue, and the pH was adjusted to 3 with 1.5 mol / L HCl. A solid precipitated out. The mixture was filtered, the filter cake was washed with acetone, collected, and dried under vacuum to give 760 mg of white solid, with a yield of 87.0%.

[0310] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.95 (s, 1H), 8.72 (d, J=10.1Hz, 2H), 8.21 (d, J= 8.7Hz,1H),7.87(d,J=8.7Hz,1H),4.79(q,J=7.2Hz,2H),1.50(t,J=7.2Hz,3H).

[0311] Synthesis of 4-amino-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-acyl chloride hydrochloride (8-3)

[0312] In a 25 mL round-bottom flask, intermediate 7-3 (760 mg, 3.03 mmol) and 5 mL of dichloromethane were added. While stirring, a 4 mol / L dioxane solution (2.33 mL, 9.30 mmol) was added, and the mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure, and the remaining solid was dissolved in 5 mL of dichloromethane. Under ice bath conditions, oxaloyl chloride (1.59 mL, 18.6 mmol) and 2 drops of DMF were added dropwise. After stirring for 5 min, the mixture was moved to room temperature and the reaction was continued for 5 h. TLC (V1) was performed. 二氯甲烷 :V 甲醇 =4:1) Monitor the reaction to ensure it is complete, remove the solvent under reduced pressure, dissolve the residue in 5 mL of dichloromethane, remove the dichloromethane under reduced pressure, repeat the operation once to remove excess oxalyl chloride, and obtain a white solid. No purification is required, and proceed directly to the next step.

[0313] Synthesis of O-ethyl-N-(4-(trifluoromethyl)benzyl)hydroxylamine (11-27)

[0314] Using 4-(trifluoromethyl)benzaldehyde 9-6 (743 mg, 4.27 mmol) and compound 10-2 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-27, yielding 410 mg of a colorless oily liquid with a yield of 47.6%.

[0315] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 7.62 (d, J = 8.0Hz, 2H), 7.51 (d, J = 8.0Hz, 2H), 4.11 (s, 2H), 3.70 (q, J = 7.0Hz, 2H), 1.14 (t, J = 7.0Hz, 3H).

[0316] Synthesis of 4-amino-N-ethoxy-1-ethyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-68)

[0317] Using intermediates 11-27 (80.0 mg, 0.392 mmol) and 8-3 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as in I-1, yielding 103.0 mg of white solid, with a yield of 61.0%.

[0318] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.47 (s, 1H), 8.33 (s, 1H), 7.84 (dd, J = 8.7, 1.9Hz, 1H), 7.78 (d, J = 8.1Hz, 2H), 7.66 (s, 1H), 7.62 (d, J=8.6Hz,2H),7.36(s,2H),5.11(s,2H),4.70(q,J=7.2Hz,2H),3.87(t,J=7.0Hz,2H),1.45(t,J=7.2Hz,3H),0.94(t,J=7.0Hz,3H).

[0319] Example 30: 4-Amino-N-(4-cyanobenzyl)-1-ethyl-N-methoxy-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-69: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0320] Synthesis of 4-((methoxyamino)methyl)benzyl nitrile (11-20)

[0321] Using 4-cyanobenzaldehyde 9-15 (560 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 235 mg of colorless oily liquid, with a yield of 33.9%.

[0322] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 6.57 (d, J = 8.5 Hz, 2H), 6.35 (d, J = 8.4 Hz, 2H), 2.79 (s, 2H), 2.14 (s, 3H).

[0323] Synthesis of 4-amino-N-(4-cyanobenzyl)-1-ethyl-N-methoxy-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-69)

[0324] Using intermediates 11-20 (69.0 mg, 0.423 mmol) and 8-3 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as in I-1, yielding 45.0 mg of white solid with a yield of 29.7%.

[0325] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.51 (s, 1H), 8.33 (s, 1H), 7.87 (d, J = 8.1Hz, 2H), 7.84 (d, J = 1.9Hz, 1H), 7.63 (d, J = 4. 7Hz,2H),7.61(d,J=4.2Hz,1H),7.32(s,2H),5.10(s,2H),4.72(q,J=7.2Hz,2H),3.63(s,3H),1.45(t,J=7.2Hz,3H).

[0326] Example 31: N-(allyloxy)-4-amino-N-((5-bromopyridin-2-yl)methyl)-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-70: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0327] Synthesis of O-allyl-N-((5-bromopyridin-2-yl)methyl)hydroxylamine (11-13)

[0328] Using compound 9-1 (794 mg, 4.27 mmol) and O-allyl hydroxylamine hydrochloride 10-5 (656 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 223 mg of colorless oily liquid, with a yield of 21.5%.

[0329] 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.61(d,J=2.1Hz,1H),8.01(dd,J=8.3,2.4Hz,1H),7.45(d,J=8.4Hz,1H),7.1 2(t,J=6.0Hz,1H),5.92-5.76(m,1H),5.22-5.05(m,2H),4.06(dt,J=5.6,1.4Hz,2H),4.00(d,J=6.0Hz,2H).

[0330] Synthesis of N-(allyloxy)-4-amino-N-((5-bromopyridin-2-yl)methyl)-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-70)

[0331] Using intermediates 11-13 (95.0 mg, 0.392 mmol) and 8-3 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as that in I-1, yielding 45.0 mg of white solid with a yield of 24.6%.

[0332] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.72 (d, J = 2.1Hz, 1H), 8.50 (s, 1H), 8.37 (s, 1H), 8.09 (d, J = 8.4Hz, 1H), 7.83 (d, J = 8.7Hz, 1H), 7.62 (d, J = 8.6Hz, 1 H),7.46(d,J=8.4Hz,1H),5.71-5.61(m,1H),5.21-5.11(m,2H),5.06(s, 2H), 4.72(q, J=7.2Hz, 2H), 4.34(d, J=6.3Hz, 2H), 1.44(t, J=7.2Hz, 3H).

[0333] Example 32: 4-Amino-N-((5-bromopyridin-2-yl)methyl)-1,3-diethyl-N-methoxy-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-71: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0334] Synthesis of N-((5-bromopyridin-2-yl)methyl)-O-methylhydroxylamine (11-1)

[0335] Using compound 9-1 (794 mg, 4.27 mmol) and O-methylhydroxylamine hydrochloride 10-1 (656 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-1, yielding 269 mg of a colorless oily liquid, with a yield of 25.8%.

[0336] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 8.63 (d, J = 2.4Hz, 1H), 7.77 (dd, J = 8.3, 2.4Hz, 1H), 7.23 (d, J = 8.3Hz, 1H), 4.10 (s, 2H), 3.52 (s, 3H).

[0337] Synthesis of 4-amino-N-((5-bromopyridin-2-yl)methyl)-1,3-diethyl-N-methoxy-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-71)

[0338] Using intermediates 11-1 (95.0 mg, 0.392 mmol) and 8-2 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 55.0 mg of white solid with a yield of 30.1%.

[0339] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.71 (s, 1H), 8.61 (s, 1H), 8.09 (d, J = 5.9Hz, 1H), 7.85 (d, J = 8.6Hz, 1H), 7.6 0(d,J=8.7Hz,1H),7.46(d,J=8.4Hz,1H),6.79(s,2H),5.05(s,2H),4.28(s,3H),3.64(s,3H),2.61(s,3H).

[0340] Example 33: N-(allyloxy)-4-amino-N-((5-bromopyridin-2-yl)methyl)-1,3-diethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-72: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0341] Synthesis of O-allyl-N-((5-bromopyridin-2-yl)methyl)hydroxylamine (11-13)

[0342] Using compound 9-1 (794 mg, 4.27 mmol) and O-allyl hydroxylamine hydrochloride 10-5 (656 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 233 mg of colorless oily liquid, with a yield of 22.5%.

[0343] 1H NMR(300MHz,DMSO-d6)δ(ppm):8.61(d,J=2.1Hz,1H),8.01(dd,J=8.3,2.4Hz,1H),7.45(d,J=8.4Hz,1H),7.1 2(t,J=6.0Hz,1H),5.92-5.76(m,1H),5.22-5.05(m,2H),4.06(dt,J=5.6,1.4Hz,2H),4.00(d,J=6.0Hz,2H).

[0344] Synthesis of N-(allyloxy)-4-amino-N-((5-bromopyridin-2-yl)methyl)-1,3-diethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-72)

[0345] Using intermediates 11-13 (95.0 mg, 0.392 mmol) and 8-2 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as that in I-1, yielding 43.0 mg of white solid with a yield of 23.5%.

[0346] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.72 (s, 1H), 8.60 (s, 1H), 8.08 (d, J = 10.8Hz, 1H), 7.83 (d, J = 10.5Hz, 1H), 7.59 (d, J = 8.7Hz, 1H), 7.4 6(d,J=8.3Hz,1H),6.76(s,2H),5.73-5.63(m,1H),5.21-5.12(m,2H),5.05(s,2H),4.36(d,J=6.2Hz,2H),4.27(s,3H),2.61(s,3H).

[0347] Example 34: 4-Amino-N-(benzyloxy)-N-((5-bromopyridin-2-yl)methyl)-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-73) R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0348] Synthesis of O-benzyl-N-((5-bromopyridin-2-yl)methyl)hydroxylamine (11-10)

[0349] Using compound 9-1 (794 mg, 4.27 mmol) and O-benzylhydroxylamine 10-3 (737 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 350 mg of colorless oily liquid, with a yield of 30.0%.

[0350] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.62 (d, J=1.8Hz, 1H), 7.99 (dd, J=8.4, 2.5Hz, 1H), 7.45 (d ,J=8.1Hz,1H),7.33-7.23(m,5H),7.22-7.13(m,1H),4.59(s,2H),4.03(d,J=4.9Hz,2H).

[0351] Synthesis of 4-amino-N-(benzyloxy)-N-((5-bromopyridin-2-yl)methyl)-1,3-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-73)

[0352] Using intermediates 11-10 (115 mg, 0.392 mmol) and 8-2 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as that in I-1, yielding 75.0 mg of white solid with a yield of 37.0%.

[0353] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.71 (d, J = 2.4Hz, 1H), 8.52 (s, 1H), 7.80 (d, J = 8.7Hz, 1H), 7.72-7.67 (m, 1H), 7.58 (d, J = 8.7Hz, 1H), 7.47 (d, J = 8 .4Hz,1H),7.25(t,J=7.0Hz,1H),7.19(t,J=7.3Hz,2H),7.07(t,J=7.3H z,2H),6.73(s,2H),5.09(s,2H),4.83(s,2H),4.11(s,3H),2.61(s,3H).

[0354] Example 35: 4-Amino-N-(4-chloro-3-methoxybenzyl)-N-methoxy-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-74: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0355] Synthesis of N-(4-chloro-3-methoxybenzyl)-O-methylhydroxylamine (11-19)

[0356] Using 4-chloro-3-methoxybenzaldehyde 9-14 (728 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 410 mg of a colorless oily liquid with a yield of 47.6%.

[0357] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.31 (d, J = 8.0Hz, 1H), 6.96 (d, J = 1.9Hz, 1H), 6.89 (dd, J = 8.1, 1.9Hz, 1H), 4.01 (s, 2H), 3.91 (s, 3H), 3.50 (s, 3H).

[0358] Synthesis of 4-amino-N-(4-chloro-3-methoxybenzyl)-N-methoxy-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-74)

[0359] Using intermediates 11-19 (80.0 mg, 0.392 mmol) and 8-3 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as in I-1, yielding 64.0 mg of white solid with a yield of 38.0%.

[0360] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.42 (s, 1H), 8.25 (s, 1H), 7.79-7.74 (m, 1H), 7.55 (d, J = 8.6Hz, 1H), 7.38 (d, J = 8.1Hz, 1H), 7.23 ( s,2H),7.11(s,1H),6.96(d,J=8.1Hz,1H),4.94(s,2H),4.64(d,J=7.2Hz,2H),3.81(s,3H),3.55(s,3H),1.38(t,J=7.2Hz,3H).

[0361] Example 36: 4-Amino-N-((5-bromopyridin-2-yl)methyl)-1-ethyl-N-methoxy-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-75: R 2 =H,R 4 =CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0362] Synthesis of N-((5-bromopyridin-2-yl)methyl)-O-methylhydroxylamine (11-1)

[0363] Using compound 9-1 (794 mg, 4.27 mmol) and O-methylhydroxylamine hydrochloride 10-1 (656 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-1, yielding 265 mg of a colorless oily liquid with a yield of 25.5%.

[0364] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 8.63 (d, J = 2.4Hz, 1H), 7.77 (dd, J = 8.3, 2.4Hz, 1H), 7.23 (d, J = 8.3Hz, 1H), 4.10 (s, 2H), 3.52 (s, 3H).

[0365] Synthesis of 4-amino-N-((5-bromopyridin-2-yl)methyl)-1-ethyl-N-methoxy-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-75)

[0366] Using intermediate 11-1 (95.0 mg, 0.392 mmol) and intermediate 8-3 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 45.0 mg of white solid with a yield of 24.6%.

[0367] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.70 (d, J = 2.4Hz, 1H), 8.53 (d, J = 1.9Hz, 1H), 8.38 (s, 1H), 8.07 (dd, J = 8.3, 2.4Hz, 1H), 7.86 (d, J = 6.8Hz,1H),7.66(d,J=3.8Hz,1H),7.44(d,J=8.4Hz,1H),5.04(s,2H),4.73(q,J=7.1Hz,2H),3.59(s,3H),1.43(t,J=7.2Hz,3H).

[0368] Example 37: 4-Amino-N-(benzyloxy)-N-((5-bromopyridin-2-yl)methyl)-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-76: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0369] Synthesis of O-benzyl-N-((5-bromopyridin-2-yl)methyl)hydroxylamine (11-10)

[0370] Using compound 9-1 (794 mg, 4.27 mmol) and O-benzylhydroxylamine 10-3 (737 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-1, yielding 350 mg of colorless oily liquid, with a yield of 30.0%.

[0371] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.62 (d, J=1.8Hz, 1H), 7.99 (dd, J=8.4, 2.5Hz, 1H), 7.45 (d ,J=8.1Hz,1H),7.33-7.23(m,5H),7.22-7.13(m,1H),4.59(s,2H),4.03(d,J=4.9Hz,2H).

[0372] Synthesis of 4-amino-N-(benzyloxy)-N-((5-bromopyridin-2-yl)methyl)-1-ethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-76)

[0373] Using intermediates 11-10 (115 mg, 0.392 mmol) and 8-3 (122 mg, 0.443 mmol) as raw materials, the preparation method was the same as that in I-1, yielding 83.0 mg of white solid with a yield of 38.0%.

[0374] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.40 (s, 1H), 8.12 (s, 1H), 8.03 (s, 1H), 7.78 (d, J = 8.3Hz, 1H), 7.51 (d, J = 8.7Hz, 1H), 7.31 (d, J = 8.6Hz, 1H) ,7.16(d,J=8.4Hz,1H),6.94-6.84(m,3H),6.74(d,J=6.8Hz,2H),4.79(s,2H),4.51(s,2H),4.28(q,J=7.1Hz,2H),1.04(d,J=7.2Hz,3H).

[0375] Example 38: 4-Amino-N-ethoxy-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-77: R 2 =H,R 4 =CH2CH3,X 1 =X 2 =CH, A ring is Synthesis of )

[0376] Synthesis of O-methyl-N-(4-(trifluoromethyl)benzyl)hydroxylamine (11-27)

[0377] Using 4-(trifluoromethyl)benzaldehyde 9-6 (728 mg, 4.27 mmol) and compound 10-1 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-27, yielding 410 mg of a colorless oily liquid with a yield of 47.6%.

[0378] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 7.62 (d, J = 8.0Hz, 2H), 7.51 (d, J = 8.0Hz, 2H), 4.11 (s, 2H), 3.70 (q, J = 7.0Hz, 2H), 1.14 (t, J = 7.0Hz, 3H).

[0379] Synthesis of 4-amino-N-ethoxy-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-77)

[0380] Using intermediates 11-27 (80.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 77.0 mg of white solid with a yield of 46.0%.

[0381] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.80 (s, 1H), 8.58 (s, 1H), 8.28 (d, J = 3.5Hz, 2H), 8.02 (d, J = 8.8Hz, 1H), 7.83 (d, J = 8. 7Hz,1H),7.77(d,J=8.1Hz,2H),7.29(s,2H),5.09(s,2H),4.34(s,3H),3.87(d,J=7.0Hz,2H),0.96(t,J=7.0Hz,3H).

[0382] Example 39: 4-Amino-N-(tert-butoxy)-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-78: R 2 =H,R 4 =C(CH3)3,X 1 =X 2 =CH, A ring is Synthesis of )

[0383] Synthesis of O-tert-butyl-N-(4-(trifluoromethyl)benzyl)hydroxylamine (11-28)

[0384] Using 4-(trifluoromethyl)benzaldehyde 9-6 (743 mg, 4.27 mmol) and compound 10-4 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-28, yielding 350 mg of a colorless oily liquid with a yield of 39.9%.

[0385] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 7.66 (d, J = 8.5 Hz, 2H), 7.55 (d, J = 7.9 Hz, 2H), 3.91 (d, J = 6.4 Hz, 2H), 1.05 (s, 9H).

[0386] Synthesis of 4-amino-N-(tert-butoxy)-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-78)

[0387] Using intermediates 11-28 (80.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 103.0 mg of white solid with a yield of 60.0%.

[0388] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.59 (s, 1H), 8.50 (s, 1H), 7.92 (d, J = 8.6Hz, 1H), 7.79 (d, J = 8. 6Hz,1H),7.75(d,J=8.1Hz,2H),7.56(d,J=8.0Hz,2H),5.05(s,2H),4.38(s,3H),1.09(s,9H).

[0389] Example 40: N-(allyloxy)-4-amino-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-79: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0390] Synthesis of O-allyl-N-(4-(trifluoromethyl)benzyl)hydroxylamine (11-29)

[0391] Using 4-(trifluoromethyl)benzaldehyde 9-6 (743 mg, 4.27 mmol) and compound 10-5 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-29, yielding 240 mg of a colorless oily liquid, with a yield of 26.2%.

[0392] 1 H NMR (300MHz, CDCl3-d) δ (ppm): 7.61 (d, J = 8.0Hz, 2H), 7.50 (d, J = 8.0Hz, 2H), 5.95-5.81 (m, 1H), 5.30-5.16 (m, 2H), 4.15 (d, J = 6.0Hz, 2H), 4.13 (s, 2H).

[0393] Synthesis of N-(allyloxy)-4-amino-1-methyl-N-(4-(trifluoromethyl)benzyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-79)

[0394] Using intermediates 11-29 (80.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 60.0 mg of white solid with a yield of 38.0%.

[0395] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.56 (s, 1H), 8.25 (s, 1H), 7.80 (d, J = 8.7Hz, 1H), 7.75 (d, J = 8.1Hz, 2H), 7.62 (d, J = 8.0Hz, 2H) ,7.57(d,J=8.7Hz,1H),7.23(s,2H),5.76-5.61(m,1H),5.21-5.11(m,2H),5.09(s,2H),4.36(d,J=6.2Hz,2H),4.31(s,3H).

[0396] Example 41: (E)-4-amino-N-((3-chloroallyl)oxy)-1-methyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-80: R 2 =H, X 1 =X 2 =CH, A ring is Synthesis of )

[0397] Synthesis of (E)-O-(3-chloroallyl)-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)hydroxylamine (11-30)

[0398] Using 6-(trifluoromethyl)pyridine-3-carboxaldehyde 9-10 (747 mg, 4.27 mmol) and compound 10-6 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that for intermediate 11-30, yielding 128 mg of a colorless oily liquid, with a yield of 14.5%.

[0399] 1 H NMR (400MHz, CDCl3-d) δ (ppm): 8.72 (s, 1H), 7.89 (d, J = 8.2Hz, 1H), 7.69 (d, J = 8.0 Hz,1H),6.20(d,J=13.4Hz,1H),5.92(m,1H),4.13(s,2H),4.07(d,J=6.8Hz,2H).

[0400] Synthesis of (E)-4-amino-N-((3-chloroallyl)oxy)-1-methyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-80)

[0401] Using intermediates 11-30 (89.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 49.0 mg of white solid with a yield of 26.3%.

[0402] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.87 (s, 1H), 8.62 (s, 1H), 8.33 (s, 1H), 8.17 (d, J = 7.7Hz, 1H), 8.01 (d, J = 8.1Hz, 1H), 7.86 (d, J = 8.3Hz ,1H),7.66(d,J=8.7Hz,1H),7.34(s,2H),6.61(d,J=13.2Hz,1H),5.96-5.78(m,1H),5.20(s,2H),4.48(d,J=6.6Hz,2H),4.41(s,3H).

[0403] Example 42: 4-Amino-N-(tert-butoxy)-1-methyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-81: R 2 =H,R 4 =C(CH3)3,X 1 =X 2 =CH, A ring is Synthesis of )

[0404] Synthesis of O-tert-butyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)hydroxylamine (11-31)

[0405] Using 6-(trifluoromethyl)pyridine-3-carboxaldehyde 9-10 (747 mg, 4.27 mmol) and compound 10-4 (500 mg, 5.99 mmol) as raw materials, the preparation method was the same as that of intermediate 11-31, yielding 128 mg of colorless oily liquid, with a yield of 14.5%.

[0406] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.70 (s, 1H), 8.03 (d, J = 8.4Hz, 1H), 7.86 (d, J = 8.0Hz, 1H), 6.46 (t, J = 6.3Hz, 1H), 3.95 (d, J = 6.2Hz, 2H), 1.03 (s, 9H).

[0407] Synthesis of 4-amino-N-(tert-butoxy)-1-methyl-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (I-81)

[0408] Using intermediates 11-31 (89.0 mg, 0.392 mmol) and 8-1 (122 mg, 0.412 mmol) as raw materials, the preparation method was the same as that of I-1, yielding 42.0 mg of white solid with a yield of 23.7%.

[0409] 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.73(s,1H),8.49(d,J=1.9Hz,1H),8.36(s,1H),8.03(d,J=5.5Hz,1H ),7.94(d,J=8.1Hz,1H),7.81(d,J=8.6Hz,1H),7.66(s,1H),4.43(s,2H),4.36(s,3H),1.11(s,9H).

[0410] Example 43: Study on the PRMT5 inhibitory activity of some compounds of the present invention

[0411] 1. Experimental reagents

[0412] PRMT5(Active Motif,Cat.No.31921),[ 3H]-SAM (PerkinElmer, Cat. No. NET155V001MC), SAM (Sigma, Cat. No. A7007-100MG), SAH (Sigma, Cat. No. A9384-25MG), MTA (Sigma, D5011-100MG), OptiPlate (384-well, white) (Perkin Elmer, Cat. No. 6007299), 384-well Flashplate (Perkin Elmer, Cat. No. SMP410A001PK).

[0413] 2. Experimental Methods

[0414] (1) Prepare 1x detection buffer (modified Tris buffer);

[0415] (2) Compound gradient dilution: The compound was transferred to the test plate in 100% DMSO via Echo, with the final concentration fraction of DMSO being 1%;

[0416] (3) PRMT5: Prepare the enzyme and [in 1x detection buffer] 3 H]-SAM mixed solution (PRMT5+MTA: enzyme prepared in 1x detection buffer, [ 3 A mixed solution of H-SAM and MTA;

[0417] (4) Prepare substrate solution in 1x detection buffer;

[0418] (5) PRMT5: Add 15 μL of enzyme and [ 3 Transfer the H]-SAM mixture to the test plate (PRMT5+MTA: add 15 μL of enzyme, [ H]-SAM mixture to the test plate, [ ... 3 (H]-SAM and MTA mixed solution transferred to the test plate);

[0419] (6) Add 10 μL of substrate solution to each well to start the reaction, and use 10 μL of 1x detection buffer as a blank control;

[0420] (7) Incubate at room temperature for 90 min;

[0421] (8) Add cold SAM to 1x detection buffer to prepare a stop solution;

[0422] (9) Add 5 μL of stop solution to each well to stop the reaction;

[0423] (10) Transfer 25 μL to the Flashplate from each well;

[0424] (11) Incubate at room temperature for at least 1 hour;

[0425] (12) Detecting the Flashplate using Microbeta;

[0426] (13) Data processing: Fit the data in Excel and calculate the suppression value using the following formula:

[0427] inh%=(Max-Signal) / (Max-Min)*100%.

[0428] 3. Experimental Results

[0429]

[0430] a A: IC 50 <10nM, B: 10nM≤IC 50 <100nM, C: 100nM≤IC 50 <1.0μM.

[0431] As can be seen from the table above, the compounds of the present invention have a synergistic inhibitory effect of PRMT5-MTA, with most of the inhibitory activity reaching nanomolar concentration levels.

[0432] Example 44: In vitro cellular SDMA activity study of some compounds of the present invention

[0433] 1. Experimental reagents

[0434] Protease inhibitors were purchased from MCE; phosphatase inhibitor II was purchased from MCE; BCA kit was purchased from Beyotime International Co., Ltd.; PVDF membrane was purchased from Millipore, USA; ECL was purchased from Novizan Biotechnology Co., Ltd.; SDMA antibody was purchased from CST Biotechnology Co., Ltd. (catalog number 13222S); β-Tubulin was purchased from CST Biotechnology Co., Ltd. (catalog number 2146S); HRP-conjugated secondary antibody was purchased from CST Biotechnology Co., Ltd.; PRMT5 antibody was purchased from Abcam Biotechnology Co., Ltd. (catalog number ab109451).

[0435] 2. Experimental Methods

[0436] Compounds were administered to HCT-116 (MTAP-null) and HCT-116 (WT) cell lines at the expected concentrations and times. Cells were washed twice with PBS and lysed for 1 h with RIPA buffer, a protease inhibitor, and a phosphatase inhibitor II. The mixture was centrifuged at 12,000 rpm for 15 min. Total protein content was quantified using a BCA kit. Proteins were diluted to a uniform concentration, added with 6× Loading Buffer, and denatured at 100 °C before being transferred to -80 °C for analysis. Equivalent proteins were subjected to 8–10% SDS-PAGE electrophoresis and then transferred to a PVDF membrane. The membrane was subsequently blocked with 5% (w / v) skim milk for 1–2 h. Proteins were detected with primary antibody and visualized with HRP-conjugated secondary antibody using ECL as the HRP substrate. Target blots were detected using a chemiluminescence system. Grayscale analysis was performed using ImageJ software.

[0437] 3. Experimental Results

[0438]

[0439] a A: IC 50 <10nM, B: 10nM≤IC 50 <100nM, C: 100nM≤IC 50 <1.0μM;

[0440] b A: 1.0 μM < IC 50 ≤50μM, B: 100nM≤IC 50 <1.0μM.

[0441] As can be seen from the table above, the compounds of the present invention have selective inhibitory effects on MTAP-deficient cells, with most of the inhibitory activity reaching nanomolar concentration levels, while also preserving the activity of PRMT5 in wild-type cells.

Claims

1. An oxime ether compound, characterized in that, Having the structure of formula (I), it also contains its pharmaceutically acceptable salt: （I）， in: R 1 Selected from the replaced C6~C 10 The aryl group, substituted C5-C9 heteroaryl group, substituted C3-C8 cycloalkyl group, wherein the substituent is selected from at least one hydrogen, halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 fluoroalkoxy, C2-C6 heterocycloalkyl, hydroxyl, amino, dimethylamino, and the heteroaryl group contains 1 to 3 heteroatoms selected from N and O; R 2 Or R 3 Each is independently selected from hydrogen; R 4 Selected from substituted C1~C6 alkyl groups, -CH2CH=CH2, ; The substituent is selected from at least one hydrogen atom; Ring A is selected from ; X 1 or X 2 Selected independently from CR 5 Or N, where R 5 Selected from hydrogen.

2. The oxime ether compound according to claim 1, characterized in that, In the structure: R 1 The substituent is selected from substituted benzene rings, substituted pyridines, substituted furans, substituted pyrimidines, and substituted cyclohexyl groups, wherein the substituent is selected from at least one hydrogen, fluorine, chlorine, bromine, cyano, trifluoromethyl, difluoromethyl, methoxy, morpholino, or dimethylamino group; R 4 Selected from -CH3, -CH2CH3, -C(CH3)3, -CH2CH=CH2, .

3. The oxime ether compound according to claim 1, characterized in that, In the structure: R 1 Selected from .

4. The oxime ether compound according to claim 1, characterized in that, Selected from any of the following compounds: 。 5. The oxime ether compound according to any one of claims 1 to 4, characterized in that, The pharmaceutically acceptable salt is a salt formed by the compound with any of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or ferulic acid.

6. A method for preparing the oxime ether compound according to any one of claims 1 to 5, characterized in that, Includes the following steps: When R 3 When it is hydrogen, (1) Compound 1 was reacted with pinacol diboronic acid ester to prepare compound 2; (2) Compound 3 was halogenated and then coupled with compound 2 to obtain compound 5; (3) Compound 5 was synthesized into compound 8 by cyclization, hydrolysis and halogenation; (4) Compound 9 is condensed and reduced with an amino ether salt, and then acylated with compound 8 to obtain compound I; ； Among them, R 1 R 2 R 4 X 1 X 2 The definition is as described in any one of claims 1 to 3; The corresponding acid is used to form a salt with compound I prepared by the above method to obtain a pharmaceutically acceptable salt of the oxime ether compound.

7. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises any one of the oxime ether compounds of claims 1 to 5 and a pharmaceutically acceptable carrier.

8. The use of any one of the oxime ether compounds of claims 1 to 5 or the pharmaceutical composition of claim 7 in the preparation of a PRMT5 inhibitor drug.

9. The application according to claim 8, characterized in that, The drug in question is an anti-tumor drug.

10. The application according to claim 9, characterized in that, The aforementioned medication is for the treatment of breast cancer, colorectal cancer, and hematologic malignancies.

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