CN117924277A

CN117924277A - Oxime ether compound and preparation method, pharmaceutical composition and application thereof

Info

Publication number: CN117924277A
Application number: CN202410079552.XA
Authority: CN
Inventors: 徐云根; 程星星; 王贞; 邹毅; 朱启华; 黄诗卉; 毛慧欢; 古宏峰
Original assignee: China Pharmaceutical University
Current assignee: China Pharmaceutical University
Priority date: 2023-06-06
Filing date: 2024-01-19
Publication date: 2024-04-26

Abstract

The invention discloses an oxime ether compound, a preparation method, a pharmaceutical composition and application thereof. The compound has a structure shown in a formula (I), and also comprises an isomer, a prodrug, a stable isotope substitution compound, a pharmaceutically acceptable salt or a mixture thereof, can effectively inhibit the activities of PRMT5 enzyme and MTAP deletion type cells, can exert the synergistic effect of PRMT5-MTA on the molecular level and the cell level, and can be used for treating various tumors; meanwhile, the method has higher selectivity to MTAP deletion type cells, does not influence the activity of PRMT5 in wild type cells, and is safer; in addition, the preparation method is simple and convenient and is easy to operate.

Description

Oxime ether compound and preparation method, pharmaceutical composition and application thereof

Technical Field

The invention relates to an oxime ether compound and a preparation method, a pharmaceutical composition and application thereof, in particular to an oxime ether compound with anti-tumor activity and a preparation method, a pharmaceutical composition and application thereof.

Background

Protein arginine methyltransferase (Protein ARGININE METHYLTRANSFERASE, PRMTs) is a S-adenosylmethionine (SAM) dependent methyltransferase that transfers methyl groups from SAM to the guanidyl nitrogen atom of histone and non-histone arginine residues. PRMTs is classified according to the arginine methylation pattern produced into forms I (PRMT 1, 2, 3, 4,6, 8), II (PRMT 5, 9) and III (PRMT 7), wherein PRMT5 is capable of catalyzing the formation of monomethylarginine and symmetrical dimethylarginine. PRMTs plays an important role in a variety of cellular functions, and can influence gene expression for signal transduction, mRNA splicing and DNA repair by histone methylation. Upregulation of PRMT5 expression occurs in a variety of human malignancies including lung cancer, ovarian cancer, colorectal cancer, breast cancer, melanoma, leukemia, and glioblastoma, and PRMT5 has also been used as an anticancer target for mantle cell lymphoma and glioblastoma.

The gene encoding methylthioadenosine phosphorylase (MTAP) is ubiquitously expressed in normal tissues. CDKN2A is one of the most frequently deleted tumor suppressor genes, MTAP is close to CDKN2A, and MTAP is frequently co-deleted with CDKN2A in tumor cells. MTAP is capable of cleaving Methyl Thioadenosine (MTA) to produce a precursor substrate for the methionine and adenine salvage pathway, so that MTA content is increased in MTAP-deficient cells, MTA competes with SAM for binding to PRMT5, resulting in decreased PRMT5 activity, and thus MTAP-deficient cells are more susceptible to further decrease in PRMT5 enzymatic activity.

The first generation PRMT5 inhibitor is mainly classified into SAM non-competitive and SAM competitive, and representative drugs include GSK-3326595, JNJ-64619178, PF-06939999, PRT543 and PRT811, which have nonselective inhibitory activity on PRMT5, are also related to patient dose-limiting thrombocytopenia, anemia and neutropenia, and have the defects of large side effect and low therapeutic index, which limit the application of the first generation PRMT5 inhibitor.

Disclosure of Invention

The invention aims to: the first object of the invention is to provide an oxime ether compound, the second object is to provide a preparation method of the compound, the third object is to provide a pharmaceutical composition containing the compound, and the fourth object is to provide an application of the compound and the pharmaceutical composition thereof.

The technical scheme is as follows: the oxime ether compound disclosed by the invention has a structure shown in a formula (I), and further comprises an isomer, a prodrug, a stable isotope substitution compound, a pharmaceutically acceptable salt or a mixture thereof:

Wherein:

R ¹ is selected from substituted C ₆～C₁₀ aryl, substituted C ₅～C₉ heteroaryl, substituted C ₃～C₈ cycloalkyl, substituted C ₂～C₆ heterocycloalkyl, substituted C ₄～C₁₂ spirocyclic or substituted C ₄～C₁₂ bridged ring, wherein said substituents are selected from at least one hydrogen, halogen, cyano, C ₁～C₃ alkyl, C ₁～C₃ haloalkyl, C ₁～C₃ alkoxy, C ₁～C₃ fluoroalkoxy, C ₂～C₆ heterocycloalkyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxy, methoxycarbonyl or nitro, said heteroaryl, heterocycloalkyl comprising 1 to 3 heteroatoms selected from N, O, S;

R ² or R ³ are each independently selected from hydrogen, substituted C ₁～C₆ alkyl or substituted C ₁～C₆ haloalkyl, wherein the substituents are selected from at least one hydrogen, halogen, cyano, C ₃～C₈ cycloalkyl, C ₂～C₆ heterocycloalkyl, hydroxy, amino, methylamino, dimethylamino, nitro, carboxy or methoxycarbonyl, the heteroaryl, heterocycloalkyl comprising 1 to 3 heteroatoms selected from N, O, S;

R ⁴ is selected from hydrogen, substituted C ₁～C₈ alkyl, substituted C ₁～C₈ haloalkyl, substituted C ₆～C₁₀ aryl, substituted C ₅～C₉ heteroaryl, substituted C ₃～C₈ cycloalkyl, substituted C ₄～C₁₂ spirocyclic, substituted C ₄～C₁₂ bridged ring, substituted vinyl, substituted ethynyl, or substituted allyl, wherein the substituents are selected from at least one hydrogen, halogen, cyano, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, C ₂～C₆ heterocycloalkyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxy, methoxycarbonyl, or nitro, the heteroaryl, heterocycloalkyl comprising 1 to 3 heteroatoms selected from N, O, S;

The A ring is selected from substituted five-membered aromatic heterocycle or five-membered heterocycloalkane, wherein the substituent is selected from at least one hydrogen, halogen, cyano, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₃～C₈ cycloalkyl, C ₂～C₆ heterocycloalkyl, C ₁～C₆ alkoxy, C ₁～C₆ acyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxyl, methoxycarbonyl or nitro, and the aromatic heterocycle, heterocycloalkane, heterocycloalkyl comprises 1 to 3 heteroatoms selected from N, O, S;

X ¹ or X ² are each independently selected from C-R ⁵ or N, wherein R ⁵ is selected from hydrogen, halogen, amino, cyano, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy or C ₁～C₆ haloalkoxy.

Preferably, in the structure:

R ¹ is selected from substituted C ₆～C₁₀ aryl, substituted C ₅～C₉ heteroaryl, substituted C ₃～C₈ cycloalkyl, substituted C ₂～C₆ heterocycloalkyl, wherein said substituents are selected from at least one hydrogen, halogen, cyano, C ₁～C₃ alkyl, C ₁～C₃ haloalkyl, C ₁～C₃ alkoxy, C ₁～C₃ fluoroalkoxy, C ₂～C₆ heterocycloalkyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxy, methoxycarbonyl, or nitro, said heteroaryl, heterocycloalkyl comprising 1 to 3 heteroatoms selected from N, O, S;

R ² or R ³ are each independently selected from hydrogen or methyl;

R ⁴ is selected from hydrogen, substituted C ₁～C₆ alkyl, substituted C ₁～C₆ haloalkyl, substituted C ₆～C₁₀ aryl, substituted C ₅～C₉ heteroaryl, substituted C ₃～C₈ cycloalkyl, substituted C ₄～C₁₂ spirocyclic group, substituted vinyl, substituted ethynyl, or substituted allyl, wherein the substituents are selected from at least one hydrogen, halogen, cyano, C ₁～C₃ alkyl, C ₁～C₃ haloalkyl, C ₁～C₃ alkoxy, C ₂～C₆ heterocycloalkyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxy, methoxycarbonyl, or nitro, the heteroaryl, heterocycloalkyl comprising 1 to 3 heteroatoms selected from N, O, S;

The A ring is selected from substituted five-membered heteroaromatic ring or five-membered heterocyclic alkane, 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 and the heterocyclic alkane comprise 1-3 heteroatoms selected from N, O, S;

X ¹ or X ² are each independently selected from C-R ⁵ or N, R ⁵ is selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, trifluoromethyl, methoxy, trifluoromethoxy or 2, 2-difluoroethyl.

Preferably, in the structure:

R ¹ 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 naphthalene rings, substituted quinolines, substituted indoles, substituted indazoles, substituted cyclopropyl, substituted cyclobutyl, substituted cyclopentyl, substituted cyclohexyl, substituted piperazines, substituted piperidines, substituted pyrroles or substituted tetrahydrofurans, wherein the substituents are selected from at least one hydrogen, fluoro, chloro, bromo, cyano, methyl, ethyl, trifluoromethyl, difluoromethyl, cyclopropyl, methoxy, trifluoromethoxy, cyclopropoxy, morpholino, piperidinyl, tetrahydropyrrolidinyl, 2- (N, N-dimethylamino) ethyl, 2- (N, N-diethylamino) ethyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxyl, methoxycarbonyl or nitro;

R ² or R ³ are each independently selected from hydrogen or methyl;

R ⁴ is selected from hydrogen, substituted C ₁～C₆ alkyl, substituted C ₁～C₆ haloalkyl, substituted C ₆～C₁₀ aryl, substituted C ₅～C₉ heteroaryl, substituted C ₃～C₈ cycloalkyl, substituted C ₄～C₁₂ spirocyclic or substituted allyl, wherein the substituents are selected from at least one hydrogen, fluoro, chloro, bromo, cyano, methyl, ethyl, trifluoromethyl, difluoromethyl, cyclopropyl, methoxy, cyclopropylmethoxy, trifluoromethoxy, acridinyl, tetrahydropyrrolidinyl, morpholinyl, piperidinyl, piperazinyl, 2- (N, N-dimethylamino) ethyl, 2- (N, N-diethylamino) ethyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxy, methoxycarbonyl or nitro, the heteroaryl comprising 1 to 3 heteroatoms selected from N, O, S;

The A ring is selected from substituted pyrrole, pyrazole, isoxazole, thiophene, thiazole, furan, dihydropyrrole or dihydrofuran, and the substituent is selected from at least one hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy and trifluoromethoxy;

X ¹ or X ² are each independently selected from C-R ⁵ or N, R ⁵ is selected from hydrogen, methyl, ethyl, trifluoromethyl, methoxy, trifluoromethoxy or 2, 2-difluoroethyl.

Preferably, in the structure:

R ¹ is selected from

R ² or R ³ are each independently selected from hydrogen or methyl;

R ⁴ is selected from -CH₃、-CH₂CH₃、-CH(CH₃)₂、-C(CH₃)₃、-CH₂CH₂OH、-CH₂CH₂CN、-CH₂CH₂F、-CH₂CH₂CHF₂、-CH₂CH₂CF₃、-CH₂CH₂OCH₃、-CH₂CH＝CH₂、

Ring A is selected from

X ¹ or X ² are each independently selected from CH, C-F, C-Cl, C-Br or N.

Specifically, the oxime ether compound is selected from any one of the following compounds:

/>

the invention designs a series of MTA synergistic PRMT5 inhibitors, which can selectively inhibit the activity of PRMT5 in MTAP-deficient cells, and simultaneously retain the activity of PRMT5 in MTAP wild cells, thereby being safer and more effective.

Preferably, the pharmaceutically acceptable salt is a salt of the compound with any one 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.

The preparation method of the oxime ether compound comprises the following steps:

When R ³ is hydrogen, the hydrogen atom,

(1) Reacting the compound 1 with pinacol biborate to obtain a compound 2;

(2) Compound 3 is halogenated and then is coupled with compound 2 to prepare compound 5;

(3) Compound 5 is cyclized, hydrolyzed and halogenated to prepare compound 8;

(4) The compound 9 and an amino ether salt compound are condensed and reduced, and then are subjected to an acylation reaction with the compound 8 to prepare a compound I;

Wherein R ¹、R²、R⁴、X¹、X² is as defined above;

And salifying the corresponding acid with the compound I prepared by the method to obtain pharmaceutically acceptable salts of the oxime ether compounds.

Specifically, in step (1), compound 2 is prepared by reacting compound 1 with pinacol biborate using a catalyst selected from [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (Pd (dppf) Cl ₂), tetrakis (triphenylphosphine) palladium (Pd (PPh ₃)₄), bis (triphenylphosphine) palladium dichloride (Pd (PPh ₃)₂Cl₂) or palladium acetate (Pd (OAc) ₂)), preferably Pd (dppf) Cl ₂), a solvent selected from dichloromethane, tetrahydrofuran, toluene, 1, 4-dioxane, ethyl acetate, acetone, N-dimethylformamide or a mixed solvent of any two, preferably 1, 4-dioxane, a base selected from potassium hydroxide, potassium carbonate, potassium acetate, sodium carbonate, preferably potassium acetate, and a reaction temperature selected from 80-120deg.C, preferably 100deg.C.

In step (2), compound 4 is prepared from compound 3 by a sandmeyer reaction using a diazotizing agent selected from the group consisting of methyl nitrite, ethyl nitrite, amyl nitrite, isoamyl nitrite, isobutyl nitrite, t-butyl nitrite, isopropyl nitrite, preferably isoamyl nitrite; the solvent and iodizing agent are selected from methyl iodide, diiodomethane, ethyl iodide, diiodoethane or mixed solvent of any two, preferably diiodomethane; the reaction temperature is selected from 60 to 100 ℃, preferably 80 ℃.

Preparing a compound 5 by reacting the compound 2 with a compound 4, wherein the solvent is selected from a mixed solvent consisting of toluene, 1, 4-dioxane, N-dimethylformamide, N-butanol and water, and preferably a mixed solvent consisting of 1, 4-dioxane and water; the base is selected from potassium hydroxide, potassium carbonate, cesium carbonate, potassium acetate, sodium carbonate, sodium bicarbonate, preferably cesium carbonate; the catalyst used is selected from the group consisting of [1,1 '-bis (diphenylphosphino) ferrocene ] palladium dichloride (Pd (dppf) Cl ₂), tetrakis (triphenylphosphine) palladium (Pd (PPh ₃)₄), bis (triphenylphosphine) palladium dichloride (Pd (PPh ₃)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) (SPhospdG 2), preferably tetrakis (triphenylphosphine) palladium (Pd (PPh ₃)₄), and the reaction temperature is selected from the group consisting of 60 to 100 ℃, preferably 80 ℃.

In the step (3), the compound 5 is subjected to an aromatization reaction to prepare a compound 6, wherein the solvent is selected from anhydrous toluene, anhydrous tetrahydrofuran, anhydrous 1, 4-dioxane, anhydrous N, N-dimethylformamide or a mixed solvent consisting of any two, and preferably the anhydrous toluene; the base 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 ℃, preferably 100 ℃.

The compound 6 is subjected to hydrolysis reaction to prepare a compound 7, wherein the base is selected from lithium hydroxide, sodium hydroxide, potassium carbonate and sodium carbonate, preferably lithium hydroxide, and the solvent is selected from methanol, ethanol, tetrahydrofuran, dioxane and water or a mixed solvent consisting of the methanol, the ethanol, the tetrahydrofuran and the water=1:1:1; the reaction temperature is selected from 50 to 90 ℃, preferably 60 ℃.

Compound 8 is prepared from compound 7 by acylation reaction, and the chlorinating agent is selected from sulfoxide chloride, phosphorus trichloride, phosphorus pentachloride and oxalyl chloride, preferably oxalyl chloride.

In the step (4), the compound 9 and the compound 10 are reacted to prepare an intermediate oxime, wherein the solvent is selected from dichloromethane, tetrahydrofuran, 1, 4-dioxane, ethyl acetate, N-dimethylformamide or a mixed solvent composed of any two, and the dichloromethane is preferred; the base used is selected from triethylamine, pyridine, N-diisopropylethylamine, 4-dimethylaminopyridine, potassium carbonate, sodium carbonate, cesium carbonate or sodium acetate, preferably pyridine. Preparing a compound 11 from the intermediate oxime through a reduction reaction, wherein the reducing agent is selected from sodium borohydride, sodium triacetoxyborohydride and sodium cyanoborohydride, and preferably sodium cyanoborohydride; the solvent is selected from methanol, ethanol, dioxane, tetrahydrofuran, glacial acetic acid or mixed solvent of any two, preferably glacial acetic acid.

The final product I is prepared by reacting compound 8 with compound 11 using a solvent selected from the group consisting of dichloromethane, ethyl acetate, tetrahydrofuran, 1, 4-dioxane, preferably tetrahydrofuran; the base used is selected from triethylamine, pyridine, N-diisopropylethylamine, preferably N, N-diisopropylethylamine.

The pharmaceutical composition provided by the invention comprises the oxime ether compound and a pharmaceutically acceptable carrier. Can be added with common medicinal adjuvants such as perfume, sweetener, liquid/solid filler, and diluent, and made into common medicinal preparation such as tablet, capsule, syrup, suspension, injection, etc.

The oxime ether compound and the pharmaceutical composition thereof are applied to the preparation of PRMT5 inhibitor drugs, and are particularly used as antitumor drugs, in particular to drugs for treating breast cancer, colorectal cancer, hematoma and the like.

The beneficial effects are that: compared with the prior art, the invention has the following remarkable advantages:

The oxime ether compound can effectively inhibit the activities of PRMT5 enzyme and MTAP-deficient cells (the IC ₅₀ value mostly reaches the nanomolar concentration level, and is optimally lower than 10 nM), can exert the synergistic effect of PRMT5-MTA at the molecular level and the cell level, and can be used for treating various tumors; meanwhile, the method has higher selectivity to MTAP deletion type cells, does not influence the activity of PRMT5 in wild type cells, and is safer; in addition, the preparation method of the compound is simple and convenient and easy to operate.

Detailed Description

The technical scheme of the invention is further described below by referring to examples.

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) 4-Amino-3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzoic acid methyl ester (2)

To a 250mL eggplant-shaped bottle were added methyl 4-amino-3-bromobenzoate 1 (4.00 g,17.4 mmol), pinacol biborate (8.83 g,34.8 mmol) and 60mL 1, 4-dioxane in this order, potassium acetate (5.12 g,52.2 mmol) and Pd (dppf) Cl ₂ (1.42 g,1.74 mmol) were added under stirring at room temperature, the reaction system was refluxed at 100℃for 18h under N ₂ protection, TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =1:1) was monitored for completion of the reaction, cooled to room temperature, suction filtered, the filter cake was washed with dichloromethane, and the filtrate was concentrated to give a semi-solid residue. Dissolving with a small amount of dichloromethane, suction filtering, collecting a filter cake, concentrating the filtrate, repeating the operation once, combining the two filter cakes, and drying to obtain 3.00g of off-white solid with the yield of 62.2%.

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).

Synthesis of 5-iodo-1-methyl-1H-pyrazole-4-carbonitrile (4-1)

To a 250mL three-necked flask, 5-amino-1-methyl-1H-pyrazole-4-carbonitrile 3-1 (5.00 g,40.9 mmol) and 90mL CH ₂I₂ were sequentially added, heated to 80℃and isoamyl nitrite (11.0 mL,81.9 mmol) was added dropwise, after the addition, heating was continued at 80℃for 1H, TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =10:1) was monitored to complete the reaction, cooled to room temperature, the reaction solution was extracted with concentrated HCl (30 mL. Times.2), concentrated HCl layers were combined, the pH of the aqueous layer was adjusted to 1 with saturated aqueous NaOH solution, extracted with ethyl acetate (30 mL. Times.2), the ethyl acetate layers were combined, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (V _{petroleum ether}:V_{Acetic acid ethyl ester} =20:1) to give 3.00g of a white solid with a yield of 31.5%.

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

Synthesis of methyl 4-amino-3- (4-cyano-1-methyl-1H-pyrazol-5-yl) benzoate (5-1)

To a 250mL eggplant-shaped bottle were added in this order intermediate 2 (3.92 mg,14.2 mmol), intermediate 4-1 (3.00 g,12.9 mmol), pd (PPh ₃)₄ (744 mg,0.644 mmol) and cesium carbonate (12.6 g,38.7 mmol), 80mL of 1, 4-dioxane and 8mL of H ₂O.N₂ for protection, the reaction was warmed to 80℃for 2H, TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =2:1) was monitored to complete the reaction, cooled to room temperature, after half of the solvent was distilled off under reduced pressure, extracted with ethyl acetate (40 mL. Times.2), the ethyl acetate layers were combined and dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (V _{petroleum ether}:V_{Acetic acid ethyl ester} =2:1+TEA) to give a yellow solid of 2.00g, yield 60.4%.

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

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

To a 25mL eggplant-shaped bottle were successively added intermediate 5-1 (1.00 g,3.90 mmol), 10mL of anhydrous toluene, and NaNH ₂ (304 mg,7.80 mmol). The reaction system was heated at 100deg.C under protection of N ₂ for 2h, TLC (V _{Dichloromethane (dichloromethane)}:V_Methanol =20:1+TEA) monitored to complete the reaction, the reaction solution was cooled to below 5deg.C, a small amount of ice water was added, the solvent was distilled off under reduced pressure, the residue was slurried with ice water, the filter cake was collected by suction filtration, and 800mg of white solid was obtained by vacuum drying in 80.0% yield.

¹H 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).

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

To a 25mL eggplant-shaped bottle, intermediate 6-1 (800 mg,3.10 mmol), 3mL THF, 3mL MeOH and 3mL H ₂ O were added sequentially, liOH (150 mg,6.24 mmol) was added under stirring, the reaction was warmed to 70℃for 2H, TLC (V _{Dichloromethane (dichloromethane)}:V_Methanol =4:1+TEA) monitored that the reaction was complete, after THF and MeOH were distilled off under reduced pressure, 6mL H ₂ O was added to the residue, pH=3 was adjusted with 1.5mol/L HCl, solids were precipitated, the filter cake was washed with acetone, the filter cake was collected, and vacuum-dried to obtain 750mg of white solid with a yield of 99.0%.

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

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

In a 25mL eggplant-shaped bottle were added intermediate 7-1 (750 mg,3.10 mmol) and 5mL of methylene chloride, and a dioxane solution (2.33 mL,9.30 mmol) of 4mol/L hydrogen chloride was added with stirring, and stirred at room temperature for 2 hours. The solvent was distilled off under reduced pressure, the remaining solid was dissolved in 5mL of dichloromethane, oxalyl chloride (1.59 mL,18.6 mmol) and 2 drops of DMF were added dropwise in sequence under ice bath conditions, after stirring for 5min, the reaction was continued at room temperature for 5h, tlc (V _{Dichloromethane (dichloromethane)}:V_Methanol =4:1) was monitored for completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved with 5mL of dichloromethane, the dichloromethane was distilled off under reduced pressure, the operation was repeated once, excess oxalyl chloride was removed to give a white solid, and the next step was directly thrown without purification.

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

To a 25mL eggplant-shaped bottle were added sequentially 5-bromopyridine-2-carbaldehyde 9-1 (794 mg,4.27 mmol), O-methylhydroxylamine hydrochloride 10-1 (500 mg,5.99 mmol) and 14mL dichloromethane, pyridine (378. Mu.L, 4.63 mmol) was added with stirring at room temperature and stirring continued for 3h, TLC (_{petroleum ether}:V_{Acetic acid ethyl ester} =10:1) was monitored to be complete, 14mL dichloromethane was added to dilute, and 14mL 1mol/L HCl and 14mL saturated brine were sequentially used to wash, dried over anhydrous sodium sulfate, suction filtered, the filtrate was collected, the solvent was distilled off under reduced pressure to obtain an oily liquid which was directly put into the next reaction without purification. To a 25mL eggplant-shaped bottle were successively added the oily liquid (360 mg,1.67 mmol) obtained above and 4mL of glacial acetic acid, naBH ₃ CN (210 mg,3.35 mmol) was slowly added with stirring, stirring was carried out at room temperature for 12h, TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =1:1) was monitored for completion of the reaction, deicing acetic acid was distilled under reduced pressure, the residue was dissolved with 8mL of 0.05mol/L HCl, then successively washed with methylene chloride (8 mL. Times.3), saturated Na ₂CO₃ solution was used to adjust pH=9, methylene chloride was extracted (8 mL. Times.3), the organic phases were combined, then successively washed with saturated Na ₂CO₃ and saturated brine, dried over anhydrous sodium sulfate, suction filtered, the filtrate was collected, and the solvent was distilled off under reduced pressure. The crude product was purified by column chromatography (V _{petroleum ether}:V_{Acetic acid ethyl ester} = 20:1) to give 160mg as a colourless oil in 17.3% yield.

¹H 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).

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

To a 25mL eggplant-shaped bottle was added sequentially intermediate 11-1 (85.0 mg, 0.390 mmol) and 1mL tetrahydrofuran, DIPEA (287. Mu.L, 1.65 mmol) and intermediate 8-1 (122 mg,0.412 mmol) dissolved in 2mL tetrahydrofuran were added sequentially under an ice bath, and after 5min, the mixture was allowed to react at room temperature for 5h. TLC (V _{Dichloromethane (dichloromethane)}:V_Methanol =20:1+tea) monitored the reaction was complete, 2mL of tetrahydrofuran was added to the reaction, the solvent was distilled off under reduced pressure, and the crude product was purified using column chromatography (V _{Dichloromethane (dichloromethane)}:V_{Ammonia methanol (7mmol/l)} =150:1) to give 60.0mg of a white solid in 34.7% yield.

¹H 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).

Example 2: 4-amino-N-methoxy-N- (4-methoxybenzyl) -1-methyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-2: r ²＝H,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using 4-methoxybenzaldehyde 9-2 (581 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, the same process as that of intermediate 11-1 gave 135mg of colorless oily liquid in 19.1% yield.

¹H 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).

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

Using intermediate 11-2 (69.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 55.0mg of a white solid with a yield of 35.8%.

¹H 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),7.33(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).

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,R⁴＝CH₂CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting with 2-bromo-4-fluorobenzaldehyde 9-3 (866 mg,4.27 mmol) and O-ethylcarboxyamine hydrochloride 10-2 (284 mg,5.99 mmol), the same process as intermediate 11-1 gave 146mg as a colorless oily liquid with a yield of 13.9%.

¹H 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).

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

Using intermediate 11-3 (97.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 64.0mg of a white solid with a yield of 34.6%.

¹H 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),7.35(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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Synthesis of N- (4-bromo-2-methoxybenzyl) -O-methylhydroxylamine (11-4)

Using 4-bromo-2-methoxybenzaldehyde 9-4 (918 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, intermediate 11-1 was prepared in the same manner to give 254mg of colorless oily liquid in 24.0% yield.

¹H 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).

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

Using intermediate 11-4 (96.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 76.0mg of a white solid with a yield of 41.0%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using 4-fluoro-3-methoxybenzaldehyde 9-5 (618 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, intermediate 11-1 was prepared in the same manner as above to obtain 164mg of colorless oily liquid with a yield of 20.7%.

¹H 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).

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

Using intermediate 11-5 (73.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as starting materials, the same procedure was followed as for I-1 to give 68.0mg of a white solid with a yield of 42.4%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (1) synthesizing O-methyl-N- (4- (trifluoromethyl) benzyl) hydroxylamine (11-6)

Starting from 4- (trifluoromethyl) benzaldehyde 9-6 (743 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as above to give 247mg as a colorless oily liquid in 28.2% yield.

¹H 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).

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

Using intermediate 11-6 (80.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 64.0mg of a white solid with a yield of 38.0%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) (Synthesis of N- ((6-fluoropyridin-3-yl) methyl) -O-methylhydroxylamine (11-7)

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

¹H 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).

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

Using intermediate 11-7 (61.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as starting materials, the same preparation as I-1 gave 50.0mg of a white solid with a yield of 33.5%.

¹H 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).

Example 8: 4-amino-N- (4-bromobenzyl) -N-methoxy-1-methyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-17: r ²＝H,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from 4-bromobenzaldehyde 9-8 (79mg, 4.27 mmol) and compound 10-1 (500 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as described above, to give 140mg of colorless oily liquid in a total yield of 15.2%.

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).

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

Using intermediate 11-8 (85.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 65.0mg of a white solid with a yield of 37.7%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

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

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).

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

Using intermediate 11-9 (66.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 70.0mg of a white solid with a yield of 45.5%.

¹H 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).

Example 10: 4-amino-N- (benzyloxy) -N- ((5-bromopyridin-2-yl) methyl) -1-methyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-24:R²＝H，/> X ¹＝X² = CH, ring a is/> ) Synthesis/>

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

Starting from compound 9-1 (794 mg,4.27 mmol) and O-benzyl hydroxylamine 10-3 (737 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as described above, to give 350mg of a colorless oily liquid, yield 30.0%.

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).

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

Using intermediate 11-10 (115 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 75.0mg of a white solid with a yield of 37.0%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is ) Is synthesized by (a)

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

Starting from 6- (trifluoromethyl) pyridine-3-carbaldehyde 9-10 (747 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as described above to give 128mg of a colorless oily liquid in 14.5% yield.

¹H 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).

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)

Using intermediate 11-11 (89.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 45.0mg of a white solid with a yield of 26.7%.

¹H 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).

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,R⁴＝C(CH₃)₃,X¹＝X² = CH, ring a is ) Is synthesized by (a)

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

Starting from compound 9-1 (794 mg,4.27 mmol) and O-tert-butylhydroxylamine 10-4 (752 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner to give 330mg of a colorless oily liquid, yield 29.8%.

¹H 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).

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)

Using intermediate 11-12 (102 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation as I-1 gave 64.0mg of a white solid with a yield of 33.8%.

¹H 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).

Example 13: n- (allyloxy) -4-amino-N- ((5-bromopyridin-2-yl) methyl) -1-methyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-32:R²＝H，/> X ¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from compound 9-1 (794 mg,4.27 mmol) and O-allylhydroxylamine hydrochloride 10-5 (650 mg,5.99 mmol), the same preparation method as intermediate 11-1 gave 265mg as a colorless oily liquid with a yield of 25.5%.

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),7.12(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).

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

Using intermediate 11-13 (95.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 45.0mg of a white solid with a yield of 24.6%.

¹H 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).

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:R²＝H，/> X ¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from compound 9-1 (794 mg,4.27 mmol) and trans-3-chloro-2-propenyl hydroxylamine 10-6 (640 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as above to give 237mg of colorless oily liquid in 20.0% yield.

¹H 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＝6.2Hz,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).

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

Using intermediate 11-14 (109 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 63.0mg of a white solid with a yield of 32.0%.

¹H 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＝8.7,1.9Hz,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).

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:R²＝H，/> X ¹＝X² = CH, ring a is ) Is synthesized by (a)

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

Starting from compound 9-1 (794 mg,4.27 mmol) and o- (4-fluorobenzyl) hydroxylamine hydrochloride 10-7 (1.06 g,5.99 mmol), the same preparation method as intermediate 11-1 gave 312mg as a colorless oily liquid in 23.5% yield.

¹H 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).

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)

Using intermediate 11-15 (122 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 85.0mg of a white solid with a yield of 40.5%.

¹H 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).

Example 16: 4-amino-N-methoxy-1-methyl-N- (4-morpholinyl) -1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-39: r ²＝H,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using 4-morpholinobenzaldehyde 9-11 (816 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, the same process as intermediate 11-1 gave 302mg of colorless oily liquid in 31.8% yield.

¹H 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).

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

Using intermediate 11-16 (87.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 70.0mg of a white solid with a yield of 40.0%.

¹H 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).

Example 17: 4-amino-N- (4-fluorobenzyl) -N-methoxy-1-methyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-40: r ²＝H,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using 4-fluorobenzaldehyde 9-12 (530 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, the same process as that of intermediate 11-1 gave 320mg of colorless oily liquid in 48.0% yield.

¹H 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).

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

Using intermediate 11-17 (61.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 65.0mg of a white solid with a yield of 44.0%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using 4- (dimethylamino) benzaldehyde 9-13 (637 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, intermediate 11-1 was prepared to give 342mg as colorless oily liquid in 44.5% yield.

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

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

Using intermediate 11-18 (71.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as starting materials, the same procedure was followed as for I-1 to give 68.0mg of a white solid with a yield of 42.9%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Synthesis of N- (4-chloro-3-methoxybenzyl) -O-methylhydroxylamine (11-19)

Using 4-chloro-3-methoxybenzaldehyde 9-14 (428 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, intermediate 11-1 was prepared in the same manner as above to obtain 410mg of colorless oily liquid in a yield of 47.6%.

¹H 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).

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

Using intermediate 11-19 (79.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 62.0mg of a white solid with a yield of 37.0%.

¹H 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).

Example 20: 4-amino-N- (4-cyanobenzyl) -N-methoxy-1-methyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-48: r ²＝H,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

Synthesis of 4- ((methoxyamino) methyl) benzonitrile (11-20)

Starting from 4-cyanobenzaldehyde 9-15 (560 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as described above, 255mg of a colorless oily liquid was obtained in 36.8% yield.

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

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

Using intermediate 11-20 (64.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 45.0mg of a white solid with a yield of 29.7%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is ) Is synthesized by (a)

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

Starting from 4- (difluoromethoxy) -3-methoxybenzaldehyde 9-16 (863 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as above to give 230mg as a colorless oily liquid in 23.1% yield.

¹H 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).

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

Using intermediate 11-21 (91.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 52.0mg of a white solid with a yield of 29.0%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

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

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

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

Using intermediate 11-22 (81.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 61.0mg of a white solid with a yield of 36.2%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) (Synthesis of N- ((5-chloropyridin-2-yl) methyl) -O-methylhydroxylamine (11-23)

Using 5-chloropyridine-2-carbaldehyde 9-18 (606 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, the same process as that of intermediate 11-1 gave 210mg of colorless oily liquid in 28.5% yield.

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).

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

Using intermediate 11-23 (68.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 51.0mg of a white solid with a yield of 32.8%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

Synthesis of 3-methoxy-4- ((methoxyamino) methyl) benzonitrile (11-24)

Using 4-cyano-3-methoxybenzaldehyde 9-19 (688 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, intermediate 11-1 was prepared in the same manner as above to give 230mg of colorless oily liquid in 28.0% yield.

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).

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

Using intermediate 11-24 (75.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 60.0mg of a white solid with a yield of 36.8%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

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

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

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

Starting from intermediate 11-25 (56.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol), the same preparation as I-1 gave 57.0mg of a white solid with a yield of 36.6%.

¹H NMR(400MHz,DMSO-d₆)δ(ppm):8.83(d,J＝1.5Hz,1H),8.63(dd,J＝7.8,1.8Hz,2H),8.32(s,1H),7.83(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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

Synthesis of 3-fluoro-4- ((methoxyamino) methyl) benzonitrile (11-26)

Starting from 4-cyano-2-fluorobenzaldehyde 9-21 (636 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner to give 230mg of a colorless oily liquid in 30.0% yield.

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

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

Using intermediate 11-26 (71.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 60.0mg of a white solid with a yield of 37.8%.

¹H 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).

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:R²＝H，/> X ¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using 4- (trifluoromethyl) benzaldehyde 9-6 (743 mg,4.27 mmol) and trans-3-chloro-2-propenyl hydroxylamine 10-6 (640 mg,5.99 mmol) as raw materials, the same process as that of intermediate 11-1 was carried out to obtain 182.0mg of colorless oily liquid with a yield of 16.1%.

¹H 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).

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

Starting from intermediate 11-27 (182.0 mg,0.687 mmol) and intermediate 8-1 (122 mg,0.412 mmol), the same procedure was followed as for I-1 to give 52.0mg of a white solid with a yield of 26.0%.

¹H 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＝8.6Hz,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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is ) Is synthesized by (a)

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

Synthesis of 5-iodo-1, 3-dimethyl-1H-pyrazole-4-carbonitrile (4-2)

To a 250mL three-necked flask, 5-amino-1, 3-dimethyl-1H-pyrazole-4-carbonitrile 3-2 (5.00 g,36.7 mmol) and 90mL CH ₂I₂ were sequentially added, and isoamyl nitrite (11.0 mL,81.9 mmol) was added dropwise, and after the addition, heating was continued for 1H at 80℃and TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =10:1) was monitored to complete the reaction, cooled to room temperature, the reaction solution was extracted with concentrated HCl (30 mL. Times.2), the concentrated HCl layer was combined, the aqueous layer pH=1 was adjusted with saturated aqueous NaOH solution, extracted with ethyl acetate (30 mL. Times.2), the ethyl acetate layer was combined, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (V _{petroleum ether}:V_{Acetic acid ethyl ester} =20:1) to give a white solid of 3.70g, yield 38.9%.

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

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

To a 250mL eggplant-shaped bottle were added sequentially intermediate 2 (4.82 g,17.5 mmol), intermediate 4-2 (3.70 g,15.9 mmol), pd (PPh ₃)₄ (892.8 mg,0.792 mmol) and cesium carbonate (15.5 g,47.6 mmol), 80mL of 1, 4-dioxane and 8mL of H ₂O.N₂ for protection, the reaction was warmed to 80℃for 2H, TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =2:1) was monitored to complete the reaction, cooled to room temperature, after half of the solvent was distilled off under reduced pressure, extracted with ethyl acetate (40 mL. Times.2), the ethyl acetate layers were combined and dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (V _{petroleum ether}:V_{Acetic acid ethyl ester} =2:1+TEA) to give a yellow solid of 2.60g, yield 78.5%.

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

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

To a 25mL eggplant-shaped bottle were added sequentially intermediate 5-2 (1.00 g,3.90 mmol), 10mL of anhydrous toluene, and NaNH ₂ (304 mg,7.80 mmol). The reaction system was heated at 100deg.C under protection of N ₂ for 2h, TLC (V _{Dichloromethane (dichloromethane)}:V_Methanol =20:1+TEA) monitored to complete the reaction, the reaction solution was cooled to below 5deg.C, a small amount of ice water was added, the solvent was distilled off under reduced pressure, the residue was slurried with ice water, filtered off with suction, the filter cake was collected, and dried under vacuum to give 830mg of white solid with a yield of 83.0%.

¹H 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).

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

To a 25mL eggplant-shaped bottle, intermediate 6-2 (830 mg,3.22 mmol), 3mL THF, 3mL MeOH and 3mL H ₂ O were added sequentially, liOH (155 mg,6.48 mmol) was added under stirring, the reaction was warmed to 70℃for 2H, TLC (V _{Dichloromethane (dichloromethane)}:V_Methanol =4:1+TEA) monitored the reaction was complete, after THF and MeOH were distilled off under reduced pressure, 6mL H ₂ O was added to the residue, pH=3 was adjusted with 1.5mol/L HCl, solids were precipitated, suction filtered, the filter cake was washed with acetone, the filter cake was collected, and vacuum dried to give 739mg of white solid with a yield of 97.5%.

¹H 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).

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

In a 25mL eggplant-shaped bottle were added intermediate 7-2 (739 mg,3.05 mmol) and 5mL of methylene chloride, and a dioxane solution (2.33 mL,9.30 mmol) of 4mol/L hydrogen chloride was added with stirring, and stirred at room temperature for 2 hours. The solvent was distilled off under reduced pressure, the remaining solid was dissolved in 5mL of dichloromethane, oxalyl chloride (1.59 mL,18.6 mmol) and 2 drops of DMF were added dropwise in sequence under ice bath conditions, after stirring for 5min, the reaction was continued at room temperature for 5h, tlc (V _{Dichloromethane (dichloromethane)}:V_Methanol =4:1) was monitored for completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved with 5mL of dichloromethane, the dichloromethane was distilled off under reduced pressure, the operation was repeated once, excess oxalyl chloride was removed to give a white solid, and the next step was directly thrown without purification.

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

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

Starting from intermediate 11-19 (80.0 mg, 0.390 mmol) and intermediate 8-2 (122 mg,0.412 mmol), the same preparation as I-1 gave 74.0mg of a white solid with a yield of 44.0%.

¹H 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).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

Synthesis of 4- ((methoxyamino) methyl) benzonitrile (11-20)

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

Using intermediate 11-20 (64.0 mg, 0.390 mmol) and intermediate 8-2 (122 mg,0.406 mmol) as raw materials, the same preparation method as I-1 gave 59.0mg of a white solid with a yield of 38.9%.

¹H 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).

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,R⁴＝CH₂CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

Synthesis of 5-iodo-1-ethyl-1H-pyrazole-4-carbonitrile (4-3)

To a 250mL three-necked flask, 5-amino-1-ethyl-1H-pyrazole-4-carbonitrile 3-3 (5.00 g,36.7 mmol) and 90mL CH ₂I₂ were sequentially added, heated to 80℃and isoamyl nitrite (11.0 mL,81.9 mmol) was added dropwise, after the addition, heating was continued at 80℃for 1H, TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =10:1) was monitored to complete the reaction, cooled to room temperature, the reaction solution was extracted with concentrated HCl (30 mL. Times.2), concentrated HCl layers were combined, the pH of the aqueous layer was adjusted to 1 with saturated aqueous NaOH solution, extracted with ethyl acetate (30 mL. Times.2), the ethyl acetate layers were combined, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (V _{petroleum ether}:V_{Acetic acid ethyl ester} =20:1) to give 3.90g of a white solid with a yield of 41.0%.

¹H 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).

Synthesis of methyl 4-amino-3- (4-cyano-1-ethyl-1H-pyrazol-5-yl) benzoate (5-3)

To a 250mL eggplant-shaped bottle were added sequentially intermediate 2 (5.10 mg,18.5 mmol), intermediate 4-3 (3.90 g,14.4 mmol), pd (PPh ₃)₄ (967.2 mg,0.837 mmol) and cesium carbonate (16.4 g,50.3 mmol), 80mL of 1, 4-dioxane and 8mL of H ₂O.N₂ for protection, the reaction was warmed to 80℃for 2H, TLC (V _{petroleum ether}:V_{Acetic acid ethyl ester} =2:1) was monitored to complete the reaction, cooled to room temperature, after half of the solvent was distilled off under reduced pressure, extracted with ethyl acetate (40 mL. Times.2), the ethyl acetate layers were combined and dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (V _{petroleum ether}:V_{Acetic acid ethyl ester} =2:1+TEA) to give 3.00g of yellow solid, yield 80.6%.

¹H 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).

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

To a 25mL eggplant-shaped bottle were added sequentially intermediate 5-3 (1.00 g,3.70 mmol), 10mL of anhydrous toluene, and NaNH ₂ (304 mg,7.80 mmol). The reaction system was heated at 100deg.C under protection of N ₂ for 2h, TLC (V _{Dichloromethane (dichloromethane)}:V_Methanol =20:1+TEA) monitored to complete the reaction, the reaction solution was cooled to below 5deg.C, a small amount of ice water was added, the solvent was distilled off under reduced pressure, the residue was slurried with ice water, the filter cake was collected by suction filtration, and the white solid 890mg was obtained by vacuum drying in 89.0% yield.

¹H 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).

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

To a 25mL eggplant-shaped bottle, intermediate 6-3 (890mg, 3.47 mmol), 3mL THF, 3mL MeOH and 3mL H ₂ O were added sequentially, liOH (150 mg,6.24 mmol) was added under stirring, the reaction was warmed to 70℃for 2H, TLC (V _{Dichloromethane (dichloromethane)}:V_Methanol =4:1+TEA) monitored that the reaction was complete, after THF and MeOH were distilled off under reduced pressure, 6mL H ₂ O was added to the residue, pH=3 was adjusted with 1.5mol/L HCl, solids were precipitated, suction filtered, the filter cake was washed with acetone, the filter cake was collected, and vacuum dried to give 760mg of white solid, yield 87.0%.

¹H 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).

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

In a 25mL eggplant-shaped bottle were added intermediate 7-3 (760 mg,3.03 mmol) and 5mL of methylene chloride, and a dioxane solution (2.33 mL,9.30 mmol) of 4mol/L hydrogen chloride was added with stirring, and stirred at room temperature for 2 hours. The solvent was distilled off under reduced pressure, the remaining solid was dissolved in 5mL of dichloromethane, oxalyl chloride (1.59 mL,18.6 mmol) and 2 drops of DMF were added dropwise in sequence under ice bath conditions, after stirring for 5min, the reaction was continued at room temperature for 5h, tlc (V _{Dichloromethane (dichloromethane)}:V_Methanol =4:1) was monitored for completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved with 5mL of dichloromethane, the dichloromethane was distilled off under reduced pressure, the operation was repeated once, excess oxalyl chloride was removed to give a white solid, and the next step was directly thrown without purification.

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

Using 4- (trifluoromethyl) benzaldehyde 9-6 (743 mg,4.27 mmol) (428 mg,4.27 mmol) and compound 10-2 (500 mg,5.99 mmol) as raw materials, the same method as that of intermediate 11-27 was used to obtain colorless oily liquid 410mg, yield 47.6%.

¹H 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).

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

Starting from intermediate 11-27 (80.0 mg, 0.390 mmol) and intermediate 8-3 (122 mg,0.443 mmol), the same preparation as for I-1 gave 103.0mg of a white solid with a yield of 61.0%.

¹H 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).

Example 30: 4-amino-N- (4-cyanobenzyl) -1-ethyl-N-methoxy-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-69: r ²＝H,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

Synthesis of 4- ((methoxyamino) methyl) benzonitrile (11-20)

Starting from 4-cyanobenzaldehyde 9-15 (560 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as described above to give 235mg of a colorless oily liquid, with a yield of 33.9%.

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

Starting from intermediate 11-20 (69.0 mg,0.423 mmol) and intermediate 8-3 (122 mg,0.443 mmol), the same procedure was followed as for I-1 to give 45.0mg of a white solid with a yield of 29.7%.

¹H 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).

Example 31: n- (allyloxy) -4-amino-N- ((5-bromopyridin-2-yl) methyl) -1-ethyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-70:R²＝H，/> X ¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using Compound 9-1 (794 mg,4.27 mmol) and O-allylhydroxylamine hydrochloride 10-5 (650 mg,5.99 mmol) as raw materials, the same preparation method as intermediate 11-1 gave 223mg as colorless oily liquid, yield 21.5%.

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

Starting from intermediate 11-13 (95.0 mg, 0.390 mmol) and intermediate 8-3 (122 mg,0.443 mmol), the same preparation as I-1 gave 45.0mg of a white solid with a yield of 24.6%.

¹H 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,1H),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).

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,R⁴＝CH3,X¹＝X² = CH, ring a is ) Is synthesized by (a)

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

Starting from compound 9-1 (794 mg,4.27 mmol) and O-methylhydroxylamine hydrochloride 10-1 (650 mg,5.99 mmol), intermediate 11-1 was prepared in the same manner as described above to give 269mg of colorless oily liquid in 25.8% yield.

¹H 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).

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)

Starting from intermediate 11-1 (95.0 mg, 0.390 mmol) and intermediate 8-2 (122 mg,0.443 mmol), the same procedure was followed as for I-1 to give 55.0mg of a white solid with a yield of 30.1%.

¹H 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),7.60(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).

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:R²＝H，/> X ¹＝X² = CH, ring a is ) Is synthesized by (a)

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

Using Compound 9-1 (794 mg,4.27 mmol) and O-allylhydroxylamine hydrochloride 10-5 (650 mg,5.99 mmol) as raw materials, the same preparation method as intermediate 11-1 gave 233mg of colorless oily liquid in 22.5% yield.

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)

Starting from intermediate 11-13 (95.0 mg, 0.390 mmol) and intermediate 8-2 (122 mg,0.443 mmol), the same preparation as I-1 gave 43.0mg of a white solid with a yield of 23.5%.

¹H 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),7.46(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).

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)R²＝H，/>X ¹＝X² = CH, ring a is) Is synthesized by (a)

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

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)

Using intermediate 11-10 (115 mg, 0.390 mmol) and intermediate 8-2 (122 mg, 0.447 mmol) as raw materials, the same process as for preparing I-1 gave 75.0mg of a white solid with a yield of 37.0%.

¹H 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＝7.3Hz,2H),6.73(s,2H),5.09(s,2H),4.83(s,2H),4.11(s,3H),2.61(s,3H).

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,R⁴＝CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

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

Starting from intermediate 11-19 (80.0 mg, 0.390 mmol) and intermediate 8-3 (122 mg,0.443 mmol), the same preparation as I-1 gave 64.0mg of a white solid with a yield of 38.0%.

¹H 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).

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,R⁴＝CH3,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from compound 9-1 (794 mg,4.27 mmol) and O-methylhydroxylamine hydrochloride 10-1 (650 mg,5.99 mmol), the same preparation method as intermediate 11-1 gave 265mg as a colorless oily liquid with a yield of 25.5%.

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

Starting from intermediate 11-1 (95.0 mg, 0.390 mmol) and intermediate 8-3 (122 mg,0.443 mmol), the same preparation as for I-1 gave 45.0mg of a white solid with a yield of 24.6%.

¹H 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).

Example 37: 4-amino-N- (benzyloxy) -N- ((5-bromopyridin-2-yl) methyl) -1-ethyl-1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-76:R²＝H，/> X ¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

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

Using intermediate 11-10 (115 mg, 0.390 mmol) and intermediate 8-3 (122 mg, 0.013 mmol) as raw materials, the same preparation as I-1 gave 83.0mg of a white solid with a yield of 38.0%.

¹H 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).

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,R⁴＝CH₂CH₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Using 4- (trifluoromethyl) benzaldehyde 9-6 (428 mg,4.27 mmol) and compound 10-1 (500 mg,5.99 mmol) as raw materials, the same process as that of intermediate 11-27 was carried out to obtain 410mg of colorless oily liquid in 47.6% yield.

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

Starting from intermediate 11-27 (80.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol), the same preparation as I-1 gave 77.0mg of a white solid with a yield of 46.0%.

¹H 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).

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,R⁴＝C(CH₃)₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from 4- (trifluoromethyl) benzaldehyde 9-6 (743 mg,4.27 mmol) and compound 10-4 (500 mg,5.99 mmol), the same process was carried out as that of intermediate 11-28 to obtain 350mg of colorless oily liquid with a yield of 39.9%.

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

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

Using intermediate 11-28 (80.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 103.0mg of a white solid with a yield of 60.0%.

¹H 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).

Example 40: n- (allyloxy) -4-amino-1-methyl-N- (4- (trifluoromethyl) benzyl) -1H-pyrazolo [4,3-c ] quinoline-8-carboxamide (I-79:R²＝H，/> X ¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from 4- (trifluoromethyl) benzaldehyde 9-6 (743 mg,4.27 mmol) and compound 10-5 (500 mg,5.99 mmol), the same process was carried out as that of intermediate 11-29 to obtain 240mg of a colorless oily liquid with a yield of 26.2%.

¹H 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).

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

Using intermediate 11-29 (80.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 60.0mg of a white solid with a yield of 38.0%.

¹H 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).

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:R²＝H， X ¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from 6- (trifluoromethyl) pyridine-3-carbaldehyde 9-10 (747 mg,4.27 mmol) and compound 10-6 (500 mg,5.99 mmol), the same preparation method as intermediate 11-30 gave 128mg of a colorless oily liquid in 14.5% yield.

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

(E) Synthesis of-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)

Starting from intermediate 11-30 (89.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol), the same preparation as I-1 gave 49.0mg of a white solid with a yield of 26.3%.

¹H 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).

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,R⁴＝C(CH₃)₃,X¹＝X² = CH, ring a is/> ) Is synthesized by (a)

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

Starting from 6- (trifluoromethyl) pyridine-3-carbaldehyde 9-10 (747 mg,4.27 mmol) and compound 10-4 (500 mg,5.99 mmol), the same process was followed as for intermediate 11-31 to give 128mg of colorless oily liquid in 14.5% yield.

¹H 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).

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)

Using intermediate 11-31 (89.0 mg, 0.390 mmol) and intermediate 8-1 (122 mg,0.412 mmol) as raw materials, the same preparation method as I-1 gave 42.0mg of a white solid with a yield of 23.7%.

¹H 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).

Example 43: PRMT5 inhibitory Activity Studies of partial Compounds of the invention

1. Experimental reagent

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).

2. Experimental method

(1) Preparing 1x detection buffer (modified Tris buffer);

(2) Compound gradient dilution: compounds were transferred to the assay plate by Echo in 100% DMSO, the final concentration fraction of DMSO was 1%;

(3) PRMT5: preparing a mixed solution of enzyme and [ ³ H ] -SAM in a 1X detection buffer (PRMT 5+ MTA: preparing a mixed solution of enzyme, [ ³ H ] -SAM and MTA in a 1X detection buffer);

(4) Preparing a substrate solution in a 1x detection buffer;

(5) PRMT5: transfer 15. Mu.L of enzyme and [ ³ H ] -SAM mixture to the test plate (PRMT 5+MTA: transfer 15. Mu.L of enzyme, [ ³ H ] -SAM and MTA mixture to the test plate);

(6) To each well 10 μl of substrate solution was added to start the reaction, 10 μl of 1x detection buffer was used as a blank;

(7) Incubating for 90min at room temperature;

(8) Adding a cold SAM into the 1x detection buffer solution to prepare a stop solution;

(9) Adding 5 mu L of stop solution into each hole, and stopping the reaction;

(10) Transfer 25 μl to Flashplate per well;

(11) Incubating at room temperature for at least 1h;

(12) Detecting the Flashplate through Microbeta;

(13) Processing data: the data were fitted in Excel and the inhibition values were calculated using the following formula:

inh％＝(Max-Signal)/(Max-Min)*100％。

3. Experimental results

^a：A：IC₅₀＜10nM,B：10nM≤IC₅₀＜100nM,C：100nM≤IC₅₀＜1.0μM。

As can be seen from the above table, the compounds of the present invention have synergistic inhibitory effects on PRMT5-MTA, with the inhibitory activity reaching nanomolar levels for the most part.

Example 44: in vitro cell SDMA Activity Studies of certain compounds of the invention

1. Experimental reagent

Protease inhibitors were purchased from MCE company; phosphatase inhibitor II was purchased from MCE company; BCA kit was purchased from bi yun tian limited; PVDF membranes were purchased from Millipore corporation, usa; ECL was purchased from south kyunovain biotechnology limited; SDMA antibodies were purchased from CST company under the designation 13222S; beta-Tubulin is available from CST company under the trade designation 2146S; HRP conjugated secondary antibodies were purchased from CST corporation; PRMT5 antibody was purchased from Abcam corporation under the accession number ab109451.

2. Experimental method

The compounds were administered at the desired concentrations and for the desired times in the HCT-116 (MTAP-null) and HCT-116 (WT) cell lines, respectively. After washing twice with PBS, cells were lysed with RIPA buffer, protease inhibitor, phosphatase inhibitor II for 1h. The mixture was centrifuged at 12000rpm for 15min. Total protein content was quantified by BCA kit, protein was diluted to uniform concentration, added 6× Loading Buffer, denatured at 100 ℃, transferred to-80 ℃ and stored for analysis. Equivalent proteins were electrophoresed by 8-10% SDS-PAGE and then transferred to PVDF membrane. Subsequently, the membrane was blocked with 5% (w/v) skimmed milk for 1-2 h. The protein was detected with primary antibody, followed by HRP conjugated secondary antibody, visualized with ECL as HRP substrate. Target blots were detected with a chemiluminescent system. Gray scale analysis was performed with Image J software.

3. Experimental results

^a：A：IC₅₀＜10nM,B：10nM≤IC₅₀＜100nM,C：100nM≤IC₅₀＜1.0μM；

^b：A：1.0μM＜IC₅₀≤50μM,B：100nM≤IC₅₀＜1.0μM。

From the above table, the compounds of the present invention have selective inhibition effect on MTAP-deleted cells, and most of the inhibition activity reaches nanomolar concentration level, and simultaneously, the activity of PRMT5 in wild type cells is well reserved.

Claims

1. An oxime ether compound characterized by having the structure of formula (I), and further comprising an isomer, a prodrug, a stable isotopically substituted compound, a pharmaceutically acceptable salt, or a mixture thereof:

Wherein:

R ² or R ³ are each independently selected from hydrogen, substituted C ₁～C₆ alkyl or substituted C ₁～C₆ haloalkyl, wherein the substituents are selected from at least one hydrogen, halogen, cyano, C ₃～C₈ cycloalkyl, C ₂～C₆ heterocycloalkyl, hydroxy, amino, methylamino, dimethylamino, nitro, carboxy or methoxycarbonyl, the heterocycloalkyl containing 1 to 3 heteroatoms selected from N, O, S;

The A ring is selected from a substituted five-membered heteroaromatic ring or five-membered heterocycloalkane, wherein the substituent is selected from at least one hydrogen, halogen, cyano, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₃～C₈ cycloalkyl, C ₂～C₆ heterocycloalkyl, C ₁～C₆ alkoxy, C ₁～C₆ acyl, hydroxy, amino, methylamino, dimethylamino, acetamido, carboxyl, methoxycarbonyl or nitro, and the heteroaromatic ring, heterocycloalkane, heterocycloalkyl comprises 1 to 3 heteroatoms selected from N, O, S;

2. The oxime ether compound according to claim 1, wherein in the structure:

R ² or R ³ are each independently selected from hydrogen or methyl;

3. The oxime ether compound according to claim 1, wherein in the structure:

R ² or R ³ are each independently selected from hydrogen or methyl;

4. The oxime ether compound according to claim 1, wherein in the structure:

R ¹ is selected from

R ² or R ³ are each independently selected from hydrogen or methyl;

Ring A is selected from

X ¹ or X ² are each independently selected from CH, C-F, C-Cl, C-Br or N.

5. Oxime ether compound according to claim 1, characterized in that it is selected from any one of the following compounds:

6. the oxime ether compound according to any one of claims 1 to 5 wherein the pharmaceutically acceptable salt is a salt of said compound with any one 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.

7. A method for producing the oxime ether compound according to any one of claims 1 to 6, comprising the steps of:

When R ³ is hydrogen, the hydrogen atom,

(1) Reacting the compound 1 with pinacol biborate to obtain a compound 2;

(3) Compound 5 is cyclized, hydrolyzed and halogenated to prepare compound 8;

Wherein R ¹、R²、R⁴、X¹、X² is as defined in any one of claims 1 to 5;

8. A pharmaceutical composition comprising the oxime ether compound according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

9. Use of an oxime ether compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 8 for the preparation of a PRMT5 inhibitor drug.

10. The use according to claim 9, wherein the medicament is an anti-tumour medicament.