Asymmetric polyfluoro-substituted 2, 4 '-biquinoline-4-amide derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicine synthesis, and discloses an asymmetric polyfluoro-substituted 2, 4 '-biquinoline-4-amide derivative as well as a preparation method and application thereof. The structural formula of the asymmetric fluorine-substituted 2, 4 '-biquinoline-4-amide derivative prepared by the invention is as shown in a general formula I: # imgabs0 #; in the formula (I), R is any one of 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 3-chloro-4-fluorophenyl, 3, 4-difluorophenyl, 2, 3, 4-trifluorophenyl and 2, 3, 4, 5-tetrafluorophenyl, and R is any one of 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 3-chloro-4-fluorophenyl, 3, 4-difluorophenyl, 2, 3, 4-trifluorophenyl and 2, 3, 4, 5-tetrafluorophenyl. And R'is cyclopropyl or dimethylaminoethyl. The asymmetric polyfluoro-substituted 2, 4 '-biquinoline-4-amide derivative has relatively strong in-vitro anti-tumor activity on human leukemia cells, human liver cancer, human pancreatic cancer and human non-small lung cancer cells, and shows relatively low cytotoxicity on normal cells; the compound can be used as an anti-tumor candidate drug with a novel structure and is mixed with an acid salt or a medicinal carrier accepted by a human body to prepare an anti-tumor drug.

Description

Technical Field

[0001] The invention belongs to the technical field of drug synthesis, and in particular relates to an asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivative and a preparation method and application thereof. Background Art

[0002] The core of drug innovation is the construction of new skeletons and their structural modification, and the creation of new skeletons based on existing drug structures and their mechanisms is the most economical and effective strategy for new drug research and development. The quinoline ring is not only a natural alkaloid such as the antimalarial drug quinine and the antitumor drug camptothecin, but also a dominant skeleton for synthetic anti-infective drugs such as fluoroquinolones. In particular, the successful launch of multiple quinoline-structured targeted antitumor drugs such as lenvatinib, anlotinib, neratinib, bosutinib, etc. and the clinical evaluation of multiple quinoline candidates in recent years have once again demonstrated the importance of quinoline as an important dominant skeleton structure in the construction of active drug molecules. However, existing studies have mainly focused on the construction and activity evaluation of single-quinoline core compounds, while there are few studies on bis-quinoline core compounds. The only studies are also limited to conjugates characterized by symmetrical structures connected by functional chains, and the construction and drugability research of asymmetric biquinoline compounds that couple two quinoline cores have not yet been reported. In addition, although the application with publication number CN118005699A discloses a novel bisquinoline cobalt complex, its structure contains a symmetrical 2,2'-biquinoline and a triquinoline structure in which a monoquinoline and cobalt form a complex, and it is a complex. Therefore, it is of great significance to develop a novel asymmetric biquinoline compound and study its anti-tumor activity. Summary of the invention

[0003] In view of the above technical problems, the present invention provides an asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivative and a preparation method and application thereof.

[0004] To achieve the above object, the technical solution of the present invention is implemented as follows:

[0005] An asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivative, the chemical structure of which is shown in Formula I:

[0006]

[0007] Wherein, R is any one of 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 3-chloro-4-fluorophenyl, 3,4-difluorophenyl, 2,3,4-trifluorophenyl and 2,3,4,5-tetrafluorophenyl; and R' is cyclopropyl or dimethylaminoethyl.

[0008] Furthermore, the above-mentioned asymmetrically polyfluorinated substituted 2,4'-biquinoline-4-amide derivative is any one of the following compounds:

[0009]

[0010] The preparation method of asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivatives comprises the following steps:

[0011] (1) Fluoroacetophenone and 5-fluoroindigo red are dissolved in a potassium hydroxide aqueous solution, and a Pfitzinger reaction is carried out to obtain a quinoline-4-carboxylic acid intermediate (such as Formula II);

[0012] (2) The quinoline-4-carboxylic acid intermediate obtained in step (1) is subjected to condensation and hydrolysis and deacidification reaction with potassium monoethyl malonate to obtain quinoline-4-ethanone (such as formula III);

[0013] (3) the quinoline-4-ethanone obtained in step (2) and 5-fluoroindigo red undergo Pfitzinger reaction in a potassium hydroxide aqueous solution to obtain 2,4'-biquinoline-4-carboxylic acid (such as formula IV);

[0014] (4) The 2,4'-biquinoline-4-carboxylic acid obtained in step (3) is reacted with cyclopropylamine or N 1 ,N 1 -dimethyl-1,2-ethylenediamine condensation to prepare asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivatives.

[0015] The structure of 5-fluoroindigo red in the above step (1) is:

[0016] In the above step (1), the fluoroacetophenone is any one of o-fluoroacetophenone, m-fluoroacetophenone, p-fluoroacetophenone, 3-chloro-4-fluoro-acetophenone, 3,4-difluoroacetophenone, 2,3,4-trifluoroacetophenone and 2,3,4,5-tetrafluoroacetophenone. The specific structural formula of the prepared quinoline-4-carboxylic acid intermediate (such as formula II) is as follows:

[0017]

[0018] The specific structural formula of quinoline-4-ethanone (such as formula III) prepared in the above step (2) is as follows:

[0019]

[0020] The specific structural formula of 2,4'-biquinoline-4-carboxylic acid (such as formula IV) prepared in the above step (3) is as follows:

[0021]

[0022]

[0023] In the above step (1), the asymmetric fluoroacetophenone is any one of o-fluoroacetophenone, m-fluoroacetophenone, p-fluoroacetophenone, 3-chloro-4-fluoro-acetophenone, 3,4-difluoroacetophenone, 2,3,4-trifluoroacetophenone and 2,3,4,5-tetrafluoroacetophenone; and the molar ratio of the fluoroacetophenone to 5-fluoroindigo red is 1:1 to 1.5.

[0024] In the above step (2), the molar ratio of the quinoline-4-carboxylic acid intermediate to the potassium salt of monoethyl malonate is 1:2-3.

[0025] In the above step (3), the molar ratio of quinoline-4-ethanone to 5-fluoroindigo red is 1:1 to 1.5.

[0026] In the above step (4), 2,4'-biquinoline-4-carboxylic acid, a condensing agent and cyclopropylamine or N 1 ,N 1 -The molar ratio of dimethyl-1,2-ethylenediamine is 1:(1~3):(1~2).

[0027] In the above step (4), the condensation agent is any one of thionyl chloride, phosphorus oxychloride, phosphorus pentoxide, polyphosphoric acid, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and carbonyldiimidazole.

[0028] Furthermore, the condensation agent is carbonyldiimidazole.

[0029] The invention discloses an application of the asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivative prepared by the invention in the preparation of anti-tumor drugs.

[0030] Furthermore, the above anti-tumor drug is a drug for treating leukemia, liver cancer, pancreatic cancer or non-small cell lung cancer.

[0031] The beneficial effects produced by the present invention are:

[0032] The present invention couples two quinoline mother nuclei to construct an asymmetric biquinoline skeleton of a novel structure, selects an amide with a hydrogen bond-accepting bifunctionality with a macromolecular target and a fluorine atom with an atomic radius close to that of hydrogen and good lipid solubility as a functional modification group of the biquinoline skeleton, and innovatively constructs a novel anti-tumor polyfluorinated substituted 2,4'-biquinoline-4-amide derivative. The IC values ​​of the polyfluorinated substituted 2,4'-biquinoline-4-amide derivatives (compounds I-1 to I-10) prepared by the present invention on four experimental cancer cell lines are 50 The values ​​all reached micromolar concentrations, showing strong anti-tumor activity, and the IC 50 The values ​​were less than micromolar concentrations, and the activity was higher than that of the control DOX or HC, especially the IC values ​​of the test compounds I-1 to I-10 against normal Vero cells. 50The values ​​are all greater than 50 micromolar concentrations, showing low cytotoxicity and potential for drug development. Therefore, according to the procedure for new drug development (first in vitro activity screening, then targeted drug development research), the compounds of the present invention have strong anti-tumor activity and low cytotoxicity, and can be mixed with acid salts acceptable to the human body or pharmaceutical carriers to prepare anti-tumor drugs. DETAILED DESCRIPTION

[0033] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] A method for preparing an asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivative, comprising the following steps:

[0035] (1) Fluoroacetophenone and 5-fluoroindigo red are dissolved in a potassium hydroxide aqueous solution, and a Pfitzinger reaction is carried out to obtain a quinoline-4-carboxylic acid intermediate (such as Formula II);

[0036] (2) The quinoline-4-carboxylic acid intermediate obtained in step (1) is subjected to condensation and hydrolysis and deacidification reaction with potassium monoethyl malonate to obtain quinoline-4-ethanone (such as formula III);

[0037] (3) the quinoline-4-ethanone obtained in step (2) and 5-fluoroindigo red undergo Pfitzinger reaction in a potassium hydroxide aqueous solution to obtain 2,4'-biquinoline-4-carboxylic acid (such as formula IV);

[0038] (4) The 2,4'-biquinoline-4-carboxylic acid obtained in step (3) is reacted with cyclopropylamine or N 1 ,N 1 -dimethyl-1,2-ethylenediamine condensation to prepare asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivatives.

[0039] In the above step (1), the fluoroacetophenone is any one of o-fluoroacetophenone, m-fluoroacetophenone, p-fluoroacetophenone, 3-chloro-4-fluoro-acetophenone, 3,4-difluoroacetophenone, 2,3,4-trifluoroacetophenone and 2,3,4,5-tetrafluoroacetophenone.

[0040] The molar ratio of fluoroacetophenone to 5-fluoroindigo red is 1:1 to 1.5.

[0041] In the above step (2), the molar ratio of the quinoline-4-carboxylic acid intermediate to the potassium salt of monoethyl malonate is 1:2 to 3. For example, the molar ratio of the quinoline-4-carboxylic acid intermediate to the potassium salt of monoethyl malonate is 1:2 to 2.5; the molar ratio of the quinoline-4-carboxylic acid intermediate to the potassium salt of monoethyl malonate is 1:2.5 to 3; the molar ratio of the quinoline-4-carboxylic acid intermediate to the potassium salt of monoethyl malonate is 1:2.4 to 3; the molar ratio of the quinoline-4-carboxylic acid intermediate to the potassium salt of monoethyl malonate is 1:2, 1:2.2, 1:2.4, 1:2.5, 1:2.6, 1:2.8 or 1:3.

[0042] In the above step (3), the molar ratio of quinoline-4-ethanone to 5-fluoroindigo red is 1:1 to 1.5. For example, the molar ratio of quinoline-4-ethanone to 5-fluoroindigo red is 1:1 to 1.3; the molar ratio of quinoline-4-ethanone to 5-fluoroindigo red is 1:1.3 to 1.5; the molar ratio of quinoline-4-ethanone to 5-fluoroindigo red is 1:1.2 to 1.5; the molar ratio of quinoline-4-ethanone to 5-fluoroindigo red is 1:1, 1:2, 1:3, 1:4 or 1:5.

[0043] In the above step (4), 2,4'-biquinoline-4-carboxylic acid, a condensing agent and cyclopropylamine or N 1 ,N 1 - dimethyl-1,2-ethylenediamine is 1:(1-3):(1-2). For example, 2,4'-biquinoline-4-carboxylic acid, condensing agent and cyclopropylamine or N 1 ,N 1 - dimethyl-1,2-ethylenediamine has a molar ratio of 1:(1.6-2):(1.7-2); 2,4'-biquinoline-4-carboxylic acid, condensing agent and cyclopropylamine or N 1 ,N 1 - dimethyl-1,2-ethylenediamine in a molar ratio of 1:(1-1.6):(1-1.7); 2,4'-biquinoline-4-carboxylic acid, condensing agent and cyclopropylamine or N 1 ,N 1 - The molar ratio of dimethyl-1,2-ethylenediamine is 1:1:1, 1:1:2, 1:3:1, 1:3:2, 1:1.6:1.7, 1:1.6:2, 1:2:1.7, 1:2:2 or 1:1.5:1.5.

[0044] In the above step (4), the condensation agent is any one of thionyl chloride, phosphorus oxychloride, phosphorus pentoxide, polyphosphoric acid, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and carbonyldiimidazole.

[0045] Furthermore, the condensing agent is carbonyldiimidazole.

[0046] Example 1

[0047] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound represented by Formula I-1) of this embodiment comprises the following steps:

[0048] (1) Synthesis of 2-(2-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1): 10.0 g (60.6 mmol) of 5-fluoroindigo red was dissolved in 10.2 g (102.0 mmol) of potassium hydroxide aqueous solution (100 mL), and 8.4 g (60.6 mmol) of 2-fluoroacetophenone was added under stirring at room temperature. The reaction mixture was stirred and refluxed for 24 hours, and an appropriate amount of activated carbon was added for decolorization for 1 hour, and filtered. The filtrate was adjusted to pH ≈ 2.0 with concentrated hydrochloric acid and allowed to stand overnight. Filtered, washed with deionized water, and dried. Recrystallized with glacial acetic acid to obtain the intermediate 2-(2-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 62.3%, mp 225~227℃. 1 H NMR(DMSO-d6,400MHz)δ:14.36(brs,1H,COOH),8.62~7.36(m,8H,Ar-H); MS(ESI)(m / z):286[M+H] + , calculate (C 16 H9F2NO2)285[M].

[0049]

[0050] (2) Synthesis of 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-1): 5 g (17.5 mol) of 2-(2-phenyl)-6-fluoro-quinoline-4-carboxylic acid was suspended in anhydrous acetonitrile (200 mL), and 5.7 g (35.2 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux for 2 hours and then kept aside for use.

[0051] Under stirring in an ice bath, 6.0 g (35.3 mmol) of potassium monoethyl malonate and 3.5 g (37.0 mmol) of anhydrous magnesium chloride were added to anhydrous tetrahydrofuran (250 mL) in sequence, and stirred for 30 minutes. Then, 45.5 g (45.0 mmol) of triethylamine was added, and the mixed reactants were stirred in an ice bath for 3 hours.

[0052] Slowly transfer the acetonitrile reaction solution to the reaction bottle of tetrahydrofuran reaction product, and the mixture of the two is refluxed for reaction for 24 hours. Recover the solvent under reduced pressure, add deionized water (200mL), and adjust the pH to 3.0 with glacial acetic acid in a water bath. Extract with dichloromethane (3×200mL), combine the organic phases, wash with water, and then wash with saturated brine. Dry with anhydrous sodium sulfate and recover the solvent to dryness. Weigh, add 3 times the weight of glacial acetic acid and 2 times the weight of concentrated hydrochloric acid, reflux and hydrolyze for 6 hours. Recover the solvent to dryness under reduced pressure, add deionized water (300mL), adjust the pH to 10.0 with 30% sodium hydroxide solution, stir well, and let it stand overnight. Filter the generated solid, wash with deionized water, and dry naturally. Recrystallize with 95% ethanol to prepare 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone shown in formula III-1. Yield 57.3%, mp 205~207℃; 1 H NMR (CDCl3, 400MHz) δ: 8.54~7.26 (m, 8H, Ar-H), 2.45 (s, 3H, COCH3); MS (ESI) (m / z): 284 [M+H] + , calculate (C 17 H 11 F2NO)283[M].

[0053]

[0054] (3) Synthesis of 2'-(2-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-1): 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (17.7 mmol) and 3.5 g (21.2 mmol) of 5-fluoroindigo red shown in Formula III-1 were dissolved in an aqueous solution (100 mL) containing 5.1 g (91.1 mmol) of potassium hydroxide. The mixed reaction solution was refluxed for 24 hours, and water (150 mL) was added. An appropriate amount of activated carbon was added and refluxed for decolorization for 1 hour. Filter, adjust pH to 2.0 with concentrated hydrochloric acid, and leave overnight. Filter, wash with water to neutralize, and dry. Recrystallize with glacial acetic acid to obtain 2'-(2-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid. Yield 68.5%, mp 245~247℃. 1 H NMR (DMSO-d6, 400MHz) δ: 14.45 (brs, 1H, COOH), 8.86 (s, 1H, 3-H), 8.56~7.32 (m, 11H, Ar-H); MS (ESI) (m / z): 431 [M+H] + , calculate (C 25 H 13 F3N2O2)430[M].

[0055]

[0056] (4) Synthesis of N-cyclopropyl-2'-(2-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-1): 2'-(2-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-1) (2.3 mmol) was suspended in anhydrous acetonitrile (20 mL), and 0.6 g (3.7 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas escaped, the mixture was heated and stirred under reflux until the solid material was completely dissolved. The mixture was allowed to stand at room temperature, and 0.26 g (4.6 mmol) of cyclopropylamine was added. The mixture was stirred at room temperature until the reaction was complete (TLC). Saturated sodium bicarbonate solution (100 mL) was added and the solid was allowed to stand to precipitate. The mixture was filtered, washed with deionized water until neutral, and dried. The mixture was recrystallized from anhydrous ethanol to obtain a light yellow crystalline substance of Formula I-1 with a yield of 56.5% and an mp of 225-227°C. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.82 (s, 1H, 3-H), 8.63-7.32 (m, 12H, Ar-H and NH), 3.67 (m, 1H, cyclopropyl-CH), 0.96-0.86 (m, 4H, CH2CH2); MS (ESI) (m / z): 470 [M+H] + , calculate (C 28 H 18 F3N3O)469[M].

[0057]

[0058] Example 2

[0059] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound shown in Formula I-2) of this embodiment comprises the following steps:

[0060] (1) Synthesis of 2-(3-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-2): 15.0 g (90.9 mmol) of 5-fluoroindigo red was dissolved in 10.2 g (102.0 mmol) of potassium hydroxide aqueous solution (100 mL), and 3-fluoroacetophenone (60.0 mmol) was added under stirring at room temperature. The reaction mixture was stirred and refluxed for 24 hours, and an appropriate amount of activated carbon was added for decolorization for 1 hour, and filtered. The filtrate was adjusted to pH ≈ 2.0 with concentrated hydrochloric acid and allowed to stand overnight. Filtered, washed with deionized water, and dried. Recrystallized with glacial acetic acid to obtain the intermediate 2-(3-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 66.8%, mp 232~234℃. 1H NMR(DMSO-d6,400MHz)δ:14.37(brs,1H,COOH),8.57~7.21(m,8H,Ar-H); MS(ESI)(m / z):286[M+H] + , calculate (C 16 H9F2NO2)285[M].

[0061]

[0062] (2) Synthesis of 2-(3-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-2): The same as step (2) in Example 1, except that 2-(2-phenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1) was replaced with 2-(3-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-2). Yield 61.7%, mp 214-216°C; 1 H NMR (CDCl3, 400MHz) δ: 8.57~7.23 (m, 8H, Ar-H), 2.46 (s, 3H, COCH3); MS (ESI) (m / z): 284 [M+H] + , calculate (C 17 H 11 F2NO)283[M].

[0063]

[0064] (3) Synthesis of 2'-(3-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-2): The same as step (3) in Example 1, except that 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (III-1) was replaced with 2-(3-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-2). Yield 73.2%, mp 246-248°C. 1 H NMR (DMSO-d6, 400MHz) δ: 14.46 (brs, 1H, COOH), 8.87 (s, 1H, 3-H), 8.53~7.35 (m, 11H, Ar-H); MS (ESI) (m / z): 431 [M+H] + , calculate (C 25 H 13 F3N2O2)430[M].

[0065]

[0066] (4) Synthesis of N-cyclopropyl-2'-(3-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-2): 1.0 g (2.3 mmol) of 2'-(3-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-2) was suspended in anhydrous acetonitrile (20 mL), and 0.65 g (4.0 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas escaped, the mixture was heated and stirred under reflux until the solid material was completely dissolved. The mixture was allowed to stand at room temperature, and 0.26 g (4.6 mmol) of cyclopropylamine was added. The mixture was stirred at room temperature until the reaction was complete (TLC). Saturated sodium bicarbonate solution (100 mL) was added and the precipitated solid was allowed to stand. The mixture was filtered, washed with deionized water until neutral, and dried. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-2 with a yield of 55.3% and mp 227-229°C. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.85 (s, 1H, 3-H), 8.65-7.34 (m, 12H, Ar-H and NH), 3.65 (m, 1H, cyclopropyl-CH), 0.97-0.86 (m, 4H, CH2CH2); MS (ESI) (m / z): 470 [M+H] + , calculate (C 28 H 18 F3N3O)469[M].

[0067]

[0068] Example 3

[0069] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound shown in Formula I-3) of this embodiment comprises the following steps:

[0070] (1) Synthesis of 2-(4-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-3): 10.0 g (60.6 mmol) of 5-fluoroindigo red was dissolved in 10.2 g (102.0 mmol) of potassium hydroxide aqueous solution (100 mL), and 4-fluoroacetophenone (60.6 mmol) was added under stirring at room temperature. The reaction mixture was stirred and refluxed for 24 hours, and an appropriate amount of activated carbon was added for decolorization for 1 hour, and filtered. The filtrate was adjusted to pH ≈ 2.0 with concentrated hydrochloric acid and allowed to stand overnight. Filtered, washed with deionized water, and dried. Recrystallized with glacial acetic acid to obtain the intermediate 2-(4-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 75.6%, mp 252~254℃. 1 H NMR(DMSO-d6,400MHz)δ:14.37(brs,1H,COOH),8.62~7.35(m,8H,Ar-H); MS(ESI)(m / z):286[M+H]+ , calculate (C 16 H9F2NO2)285[M].

[0071]

[0072] (2) Synthesis of 2-(4-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-3): The same as step (2) in Example 1, except that 2-(2-phenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1) was replaced with 2-(4-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-3). Yield 64.8%, mp 232-234°C; 1 H NMR (CDCl3, 400MHz) δ: 8.61~7.27 (m, 8H, Ar-H), 2.47 (s, 3H, COCH3); MS (ESI) (m / z): 284 [M+H] + , calculate (C 17 H 11 F2NO)283[M].

[0073]

[0074] (3) Synthesis of 2'-(4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-3): The same as step (3) in Example 1, except that 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (III-1) was replaced with 2-(4-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-3). Yield 76.3%, mp 254-256°C. 1 H NMR (DMSO-d6, 400MHz) δ: 14.47 (brs, 1H, COOH), 8.86 (s, 1H, 3-H), 8.56~7.37 (m, 11H, Ar-H); MS (ESI) (m / z): 431 [M+H] + , calculate (C 25 H 13 F3N2O2)430[M].

[0075]

[0076] (4) Synthesis of N-cyclopropyl-2'-(4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-3): 1.0 g (2.3 mmol) of 2'-(4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-3) was suspended in anhydrous acetonitrile (20 mL), and 0.75 g (4.6 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas escaped, the mixture was heated and stirred under reflux until the solid material was completely dissolved. The mixture was allowed to stand at room temperature, and 0.23 g (4.0 mmol) of cyclopropylamine was added. The mixture was stirred at room temperature until the reaction was complete (TLC). Saturated sodium bicarbonate solution (100 mL) was added and the precipitated solid was allowed to stand. The mixture was filtered, washed with deionized water until neutral, and dried. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-3 with a yield of 62.3% and mp 234-236°C. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.87 (s, 1H, 3-H), 8.68-7.36 (m, 12H, Ar-H and NH), 3.68 (m, 1H, cyclopropyl-CH), 1.02-0.87 (m, 4H, CH2CH2); MS (ESI) (m / z): 470 [M+H] + , calculate (C 28 H 18 F3N3O)469[M].

[0077]

[0078] Example 4

[0079] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound represented by Formula I-4) of this embodiment comprises the following steps:

[0080] (1) The synthesis of 2-(2-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1) is the same as in Example 1.

[0081] (2) The synthesis of 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-1) is the same as in Example 1.

[0082] (3) The synthesis of 2'-(2-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-1) is the same as in Example 1.

[0083] (4) Synthesis of N-(2-dimethylamino-1-ethyl)-2'-(2-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-4): 1.0 g (2.2 mmol) of 2'-(2-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-1) was suspended in anhydrous acetonitrile (20 mL), and 0.65 g (4.0 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux until the solid material was completely dissolved. After standing at room temperature, N 1 , N 1 -Dimethyl-1,2-ethylenediamine 0.35 g (4.0 mmol), stir at room temperature until the reaction is complete (TLC). Add saturated sodium bicarbonate solution (100 mL), let stand to precipitate solid. Filter, wash with deionized water until neutral, and dry. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-4, with a yield of 51.7%, mp 221~223℃. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.82 (s, 1H, 3-H), 8.62-7.36 (m, 12H, Ar-H and NH), 3.28-3.16 (m, 2H, CH2), 2.42-2.36 (m, 2H, CH2), 2.26, 2.24 (s, 6H, N(CH3)2); MS (ESI) (m / z): 501 [M+H] + , calculate (C 29 H 23 F3N4O)500[M].

[0084]

[0085] Example 5

[0086] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound represented by Formula I-5) of this embodiment comprises the following steps:

[0087] (1) The synthesis of 2-(3-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-2) is the same as in Example 2.

[0088] (2) The synthesis of 2-(3-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-2) is the same as in Example 2.

[0089] (3) The synthesis of 2'-(3-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-2) is the same as in Example 2.

[0090] (4) Synthesis of N-(2-dimethylamino-1-ethyl)-2'-(3-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-5): 1.0 g (2.3 mmol) of 2'-(3-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-2) was suspended in anhydrous acetonitrile (20 mL), and 0.70 g (4.3 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux until the solid material was completely dissolved. After standing at room temperature, N 1 , N 1 -Dimethyl-1,2-ethylenediamine 0.35 g (4.0 mmol), stir at room temperature until the reaction is complete (TLC). Add saturated sodium bicarbonate solution (100 mL), let stand to precipitate solid. Filter, wash with deionized water until neutral, and dry. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-5, with a yield of 57.6%, mp 232~234℃. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.85 (s, 1H, 3-H), 8.67-7.38 (m, 12H, Ar-H and NH), 3.32-3.17 (m, 2H, CH2), 2.44-2.38 (m, 2H, CH2), 2.28, 2.25 (s, 6H, N(CH3)2); MS (ESI) (m / z): 501 [M+H] + , calculate (C 29 H 23 F3N4O)500[M].

[0091]

[0092] Example 6

[0093] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound represented by Formula I-6) of this embodiment comprises the following steps:

[0094] (1) The synthesis of 2-(4-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-3) is the same as in Example 3.

[0095] (2) The synthesis of 2-(4-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-3) is the same as in Example 3.

[0096] (3) The synthesis of 2'-(4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-3) is the same as in Example 3.

[0097] (4) Synthesis of N-(2-dimethylamino-1-ethyl)-2'-(4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-6): 1.0 g (2.3 mmol) of 2'-(4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-3) was suspended in anhydrous acetonitrile (20 mL), and 0.73 g (4.5 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux until the solid material was completely dissolved. After standing at room temperature, N 1 , N 1 -Dimethyl-1,2-ethylenediamine 0.40 g (4.5 mmol), stir at room temperature until the reaction is complete (TLC). Add saturated sodium bicarbonate solution (100 mL), let stand to precipitate solid. Filter, wash with deionized water until neutral, and dry. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-6, with a yield of 60.3%, mp 238~240℃. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.87 (s, 1H, 3-H), 8.72-7.43 (m, 12H, Ar-H and NH), 3.35-3.22 (m, 2H, CH2), 2.42-2.36 (m, 2H, CH2), 2.26, 2.24 (s, 6H, N(CH3)2); MS (ESI) (m / z): 501 [M+H] + , calculate (C 29 H 23 F3N4O)500[M].

[0098]

[0099] Example 7

[0100] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound shown in Formula I-7) of this embodiment comprises the following steps:

[0101] (1) Synthesis of 2-(3-chloro-4-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-4): 10.0 g (60.6 mmol) of 5-fluoroindigo red was dissolved in 10.2 g (102.0 mmol) of potassium hydroxide aqueous solution (100 mL), and 3-chloro-4-fluoro-acetophenone (60.6 mmol) was added under stirring at room temperature. The reaction mixture was stirred and refluxed for 24 hours, and an appropriate amount of activated carbon was added for decolorization for 1 hour, and filtered. The filtrate was adjusted to pH ≈ 2.0 with concentrated hydrochloric acid and allowed to stand overnight. Filtered, washed with deionized water, and dried. Recrystallized with glacial acetic acid to obtain the intermediate 2-(3-chloro-4-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 64.7%, mp 256~258℃. 1H NMR(DMSO-d6,400MHz)δ:14.36(brs,1H,COOH),8.58~7.36(m,7H,Ar-H); MS(ESI)(m / z):320[M+H] + ( 35 Cl) and 322

[0102] [M+H] + ( 37 Cl), calculate (C 16 H8ClF2NO2)319[M].

[0103]

[0104] (2) Synthesis of 2-(3-chloro-4-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-4): The same as step (2) in Example 1, except that 2-(2-phenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1) was replaced with 2-(3-chloro-4-fluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 72.6%, mp 242-244°C; 1 H NMR (CDCl3, 400MHz) δ: 8.65~7.42 (m, 7H, Ar-H), 2.48 (s, 3H, COCH3); MS (ESI) (m / z): 318 [M+H] + ( 35 Cl), 320[M+H] + ( 37 Cl), calculate (C 17 H 10 ClF2NO)317[M].

[0105]

[0106] (3) Synthesis of 2'-(3-chloro-4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-4): The same as step (3) in Example 1, except that 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (III-1) was replaced with 2-(3-chloro-4-fluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-4). Yield 74.6%, mp 257-259°C. 1 H NMR (DMSO-d6, 400MHz) δ: 14.45 (brs, 1H, COOH), 8.86 (s, 1H, 3-H), 8.53~7.33 (m, 10H, Ar-H); MS (ESI) (m / z): 464 [M+H] + , calculate (C 25 H 12ClF3N2O2)464[M].

[0107]

[0108] (4) Synthesis of N-(2-dimethylamino-1-ethyl)-2'-(3-chloro-4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-7): 1.0 g (2.2 mmol) of 2'-(3-chloro-4-fluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-4) was suspended in anhydrous acetonitrile (20 mL), and 0.66 g (4.1 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux until the solid material was completely dissolved. After standing at room temperature, N 1 , N 1 -Dimethyl-1,2-ethylenediamine 0.35 g (4.0 mmol), stir at room temperature until the reaction is complete (TLC). Add saturated sodium bicarbonate solution (100 mL), let stand to precipitate solid. Filter, wash with deionized water until neutral, and dry. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-7, with a yield of 72.5%, mp 251-253°C. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.88 (s, 1H, 3-H), 8.75-7.46 (m, 11H, Ar-H and NH), 3.38-3.26 (m, 2H, CH2), 2.45-2.37 (m, 2H, CH2), 2.32, 2.28 (s, 6H, N(CH3)2); MS (ESI) (m / z): 535 [M+H] + , calculate (C 29 H 22 ClF3N4O)534[M].

[0109]

[0110] Example 8

[0111] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound shown in Formula I-8) of this embodiment comprises the following steps:

[0112] (1) Synthesis of 2-(3,4-difluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-5): 10.0 g (60.6 mmol) of 5-fluoroindigo red was dissolved in 10.2 g (102.0 mmol) of potassium hydroxide aqueous solution (100 mL), and 3,4-difluoroacetophenone (60.6 mmol) was added under stirring at room temperature. The reaction mixture was stirred and refluxed for 24 hours, and an appropriate amount of activated carbon was added for decolorization for 1 hour, and filtered. The filtrate was adjusted to pH ≈ 2.0 with concentrated hydrochloric acid and allowed to stand overnight. Filtered, washed with deionized water, and dried. Recrystallized with glacial acetic acid to obtain the intermediate 2-(3,4-difluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 67.3%, mp 254~256℃. 1 H NMR (DMSO-d6, 400 MHz) δ: 14.42 (brs, 1H, COOH), 8.72-7.45 (m, 7H, Ar-H); MS (ESI) (m / z): 304 [M+H], calculated (C 16 H8F3NO2)303[M].

[0113]

[0114] (2) Synthesis of 2-(3,4-difluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-5): The same as step (2) in Example 1, the only difference is that 2-(2-phenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1) is replaced by 2-(3,4-difluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-5). Yield 53.2%, mp 231~232℃; 1 H NMR (CDCl3, 400MHz) δ: 8.63~7.41 (m, 7H, Ar-H), 2.47 (s, 3H, COCH3); MS (ESI) (m / z): 302 [M+H] + , calculate (C 17 H 10 F3NO)301[M].

[0115]

[0116] (3) Synthesis of 2'-(3,4-difluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-5): The same as step (3) in Example 1, except that 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (III-1) was replaced with 2-(3,4-difluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-5). Yield 68.4%, mp 253-255°C. 1H NMR (DMSO-d6, 400MHz) δ: 14.47 (brs, 1H, COOH), 8.88 (s, 1H, 3-H), 8.62~7.36 (m, 10H, Ar-H); MS (ESI) (m / z): 449 [M+H] + , calculate (C 25 H 12 F4N2O2)448[M].

[0117]

[0118] (4) Synthesis of N-(2-dimethylamino-1-ethyl)-2'-(3,4-difluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-8): 1.0 g (2.2 mmol) of 2'-(3,4-difluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-5) was suspended in anhydrous acetonitrile (20 mL), and 0.66 g (4.1 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux until the solid material was completely dissolved. After standing at room temperature, N 1 , N 1 -Dimethyl-1,2-ethylenediamine 0.35 g (4.0 mmol), stir at room temperature until the reaction is complete (TLC). Add saturated sodium bicarbonate solution (100 mL), let stand to precipitate solid. Filter, wash with deionized water until neutral, and dry. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-8, with a yield of 64.7%, mp 232-234°C. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.90 (s, 1H, 3-H), 8.78-7.45 (m, 11H, Ar-H and NH), 3.42-3.36 (m, 2H, CH2), 2.46-2.36 (m, 2H, CH2), 2.36, 2.32 (s, 6H, N(CH3)2); MS (ESI) (m / z): 519 [M+H] + , calculate (C 29 H 22 F4N4O)518[M].

[0119] Example 9

[0120] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound shown in Formula I-9) of this embodiment comprises the following steps:

[0121] (1) Synthesis of 2-(2,3,4-trifluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-6): 10.0 g (60.6 mmol) of 5-fluoroindigo red was dissolved in 10.2 g (102.0 mmol) of potassium hydroxide aqueous solution (100 mL), and 2,3,4-trifluoroacetophenone (60.6 mmol) was added under stirring at room temperature. The reaction mixture was stirred and refluxed for 24 hours, and an appropriate amount of activated carbon was added for decolorization for 1 hour, and filtered. The filtrate was adjusted to pH ≈ 2.0 with concentrated hydrochloric acid and allowed to stand overnight. Filtered, washed with deionized water, and dried. Recrystallized with glacial acetic acid to obtain the intermediate 2-(2,3,4-trifluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 56.5%, mp 243~245℃. 1 H NMR (DMSO-d6, 400 MHz) δ: 14.36 (brs, 1H, COOH), 8.68-7.25 (m, 6H, Ar-H); MS (ESI) (m / z): 322 [M+H], calculated (C 16 H7F4NO2)321[M].

[0122]

[0123] (2) Synthesis of 2-(2,3,4-trifluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-6): The same as step (2) in Example 1, the only difference is that 2-(2-phenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1) is replaced by 2-(2,3,4-trifluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-6). Yield 47.6%, mp 226~228℃; 1 H NMR (CDCl3, 400MHz) δ: 8.65~7.37 (m, 7H, Ar-H), 2.48 (s, 3H, COCH3); MS (ESI) (m / z): 320 [M+H] + , calculate (C 17 H9F4NO)319[M].

[0124]

[0125] (3) Synthesis of 2'-(2,3,4-trifluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-6): The same as step (3) in Example 1, except that 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (III-1) was replaced with 2-(2,3,4-trifluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-6). Yield 68.4%, mp 255-257°C. 1H NMR (DMSO-d6, 400MHz) δ: 14.45 (brs, 1H, COOH), 8.87 (s, 1H, 3-H), 8.60 ~ 7.35 (m, 9H, Ar-H); MS (ESI) (m / z): 467

[0126] [M+H] + , calculate (C 25 H 11 F5N2O2)466[M].

[0127]

[0128] (4) Synthesis of N-(2-dimethylamino-1-ethyl)-2'-(2,3,4-trifluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-9): 1.0 g (2.1 mmol) of 2'-(2,3,4-trifluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-6) was suspended in anhydrous acetonitrile (20 mL), and 0.65 g (4.0 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux until the solid material was completely dissolved. After standing at room temperature, N 1 , N 1 -Dimethyl-1,2-ethylenediamine 0.35 g (4.0 mmol), stir at room temperature until the reaction is complete (TLC). Add saturated sodium bicarbonate solution (100 mL), let stand to precipitate solid. Filter, wash with deionized water until neutral, and dry. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-9, with a yield of 65.8%, mp 236-238°C. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.92 (s, 1H, 3-H), 8.82-7.43 (m, 10H, Ar-H and NH), 3.44-3.35 (m, 2H, CH2), 2.46-2.35 (m, 2H, CH2), 2.34, 2.32 (s, 6H, N(CH3)2); MS (ESI) (m / z): 537 [M+H] + , calculate (C 29 H 21 F5N4O)536[M].

[0129]

[0130] Example 10

[0131] The preparation method of the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative (compound shown in Formula I-10) of this embodiment comprises the following steps:

[0132] (1) Synthesis of 3-(2,3,4,5-tetrafluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-7): 10.0 g (60.6 mmol) of 5-fluoroindigo red was dissolved in 10.2 g (102.0 mmol) of potassium hydroxide aqueous solution (100 mL), and 2,3,4,5-tetrafluoroacetophenone (60.6 mmol) was added under stirring at room temperature. The reaction mixture was stirred and refluxed for 24 hours, and an appropriate amount of activated carbon was added for decolorization for 1 hour, and filtered. The filtrate was adjusted to pH ≈ 2.0 with concentrated hydrochloric acid and allowed to stand overnight. Filtered, washed with deionized water, and dried. Recrystallized with glacial acetic acid to obtain the intermediate 3-(2,3,4,5-tetrafluorophenyl)-6-fluoro-quinoline-4-carboxylic acid. Yield 62.3%, mp 238~240℃. 1 HNMR (DMSO-d6, 400MHz)δ:14.38(brs,1H,COOH),8.75~7.30(m,5H,Ar-H);MS(ESI)(m / z):340[M+H],calculated(C 16 H6F5NO2)339[M].

[0133]

[0134] (2) Synthesis of 2-(2,3,4,5-tetrafluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-7): The same as step (2) in Example 1, the only difference is that 2-(2-phenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-1) is replaced by 3-(2,3,4,5-tetrafluorophenyl)-6-fluoro-quinoline-4-carboxylic acid (Formula II-7). Yield 47.6%, mp 223~225℃; 1 H NMR (CDCl3, 400MHz) δ: 8.66~7.45 (m, 7H, Ar-H), 2.47 (s, 3H, COCH3); MS (ESI) (m / z): 339 [M+H] + , calculate (C 17 H8F5NO)338[M].

[0135]

[0136] (3) Synthesis of 2'-(2,3,4,5-tetrafluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-7): The same as step (3) in Example 1, except that 2-(2-fluorophenyl)-6-fluoro-quinoline-4-ethanone (III-1) was replaced with 2-(2,3,4,5-tetrafluorophenyl)-6-fluoro-quinoline-4-ethanone (Formula III-7). Yield 53.6%, mp 246-248°C. 1HNMR (DMSO-d6, 400MHz) δ: 14.46 (brs, 1H, COOH), 8.86 (s, 1H, 3-H), 8.65~7.37 (m, 8H, Ar-H); MS (ESI) (m / z): 485

[0137] [M+H] + , calculate (C 25 H 10 F6N2O2)484[M].

[0138]

[0139] (4) Synthesis of N-(2-dimethylamino-1-ethyl)-2'-(2,3,4,5-tetrafluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-amide (Formula I-10): 1.0 g (2.1 mmol) of 2'-(2,3,4,5-tetrafluorophenyl)-6,6'-difluoro-2,4'-biquinoline-4-carboxylic acid (Formula IV-7) was suspended in anhydrous acetonitrile (20 mL), and 0.60 g (3.7 mmol) of carbonyldiimidazole (CDI) was slowly added under stirring. After no gas was released, the mixture was heated and stirred under reflux until the solid material was completely dissolved. After standing at room temperature, N 1 , N 1 -Dimethyl-1,2-ethylenediamine 0.32g (3.8mmol), stir at room temperature until the reaction is complete (TLC). Add saturated sodium bicarbonate solution (100mL), let stand to precipitate solid. Filter, wash with deionized water until neutral, and dry. Recrystallize with anhydrous ethanol to obtain light yellow crystals of formula I-10, with a yield of 60.2%, mp 232~234℃. 1 H NMR (DMSO-d6, 400 MHz) δ: 8.91 (s, 1H, 3-H), 8.84-7.42 (m, 9H, Ar-H and NH), 3.42-3.37 (m, 2H, CH2), 2.43-2.36 (m, 2H, CH2), 2.36, 2.33 (s, 6H, N(CH3)2); MS (ESI) (m / z): 555 [M+H] + , calculate (C 29 H 20 F6N4O)554[M].

[0140]

[0141] Application Examples

[0142] Antitumor activity experiment: In vitro antitumor activity determination of the asymmetric polyfluorosubstituted 2,4'-biquinoline-4-amide derivatives prepared in Examples 1-10

[0143] (1) Test samples

[0144] The 10 new asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivatives provided in Examples 1-10, the broad-spectrum antitumor drug doxorubicin (DOX) and the quinoline antitumor drug hydroxycamptothecin (HC) were used as test samples, a total of 12 types, of which DOX and HC were used as controls, and the samples in Examples 1-10 were used as experimental groups.

[0145] The experimental cancer cell lines were leukemia cell line (HL60), human liver cancer cell line (SMMC-7721), human pancreatic cancer cell line (Capan-1) and human non-small cell lung cancer cell (A549). The four cancer cell lines were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. The normal cell line was African green monkey kidney cell (Vero), which was purchased from Shanghai Tongpai Biotechnology Co., Ltd.

[0146] (2) Measurement method

[0147] The above 12 test compounds were dissolved in dimethyl sulfoxide (DMSO) and prepared into 1.0×10 -2 mol·L -1 The stock solution was diluted with RPMI-1640 culture medium containing 10% calf serum to form seven concentration gradients (0.01, 0.1, 1.0, 5.0, 10.0, 50.0, 100 μmol·L -1 ) of the working fluid to be tested.

[0148] Human leukemia cell line HL60, human hepatoma cell line SMMC-7721, human pancreatic cancer cell line Capan-1, human non-small cell lung cancer cell A549 and African green kidney cell Vero in logarithmic growth phase were seeded in 96-well plates at 6 000 cells per well, and then the test solutions with 7 concentration gradients were added respectively. After 48 h of culture, 5 g·L -1 Add 10 μL of MTT (thiazolyl blue) solution and continue culturing for another 4 h; discard the supernatant, add 150 μL of DMSO to each well, place in a 37°C shaker to dissolve the crystals, and measure the absorbance (OD) value at a wavelength of 570 nm using an enzyme marker.

[0149] The inhibition rate of different concentrations of the tested drug solution on cancer cells was calculated according to the formula: cell inhibition rate = [(1- OD value of the experimental group) / OD value of the control group] × 100%; then the negative logarithmic value of each concentration of the tested drug solution was used to perform a linear regression on the cancer cell inhibition rate corresponding to each concentration to obtain a dose-effect equation, thereby calculating the half-maximal inhibition concentration (IC) of the tested drug on the experimental cancer cells. 50 ) value; each data was measured 3 times in parallel and the average value was calculated.

[0150] The half inhibitory concentration (IC) of 4 cancer cell lines and 1 normal cell line, a total of 5 experimental cell lines 50 The results of the determination of the ) values ​​are shown in Table 1.

[0151] Table 1 Antitumor activity (IC 50 )(n=3)

[0152]

[0153] The results in Table 1 show that the IC values ​​of the test compounds I-1 to I-10 against the four experimental cancer cell lines are 50 The values ​​all reached micromolar concentrations, showing strong anti-tumor activity, and the IC 50 The values ​​were less than micromolar concentrations, and the activity was higher than that of the control DOX or HC, especially the IC values ​​of the test compounds I-1 to I-10 against normal Vero cells. 50 The values ​​are all greater than 50 micromolar concentrations, showing low cytotoxicity and potential for drug development. Therefore, according to the procedure for new drug development (first in vitro activity screening, then targeted drug development research), the compounds of the present invention have strong anti-tumor activity and low cytotoxicity, and can be mixed with acid salts acceptable to the human body or pharmaceutical carriers to prepare anti-tumor drugs.

[0154] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An asymmetrically polyfluorinated substituted 2,4'-biquinoline-4-amide derivative, characterized in that: The structural formula of the polyfluoro-substituted 2,4'-biquinoline-4-amide derivative is shown in Formula I: Wherein, R is any one of 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 3-chloro-4-fluorophenyl, 3,4-difluorophenyl, 2,3,4-trifluorophenyl and 2,3,4,5-tetrafluorophenyl; and R' is cyclopropyl or dimethylaminoethyl.

2. The asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 1, characterized in that: The asymmetric polyfluorinated substituted 2,4'-biquinoline-4-amide derivative is any one of the following compounds:

3. The method for preparing the asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 1, characterized in that: Here are the steps: (1) Fluoroacetophenone and 5-fluoroindigo red undergo Pfitzinger reaction in a potassium hydroxide aqueous solution to obtain a quinoline-4-carboxylic acid intermediate; (2) the quinoline-4-carboxylic acid intermediate obtained in step (1) is subjected to condensation and hydrolysis and deacidification reaction with monoethyl malonate potassium salt to obtain quinoline-4-ethanone; (3) the quinoline-4-ethanone obtained in step (2) and 5-fluoroindigo red are reacted in a potassium hydroxide aqueous solution to produce 2,4'-biquinoline-4-carboxylic acid by Pfitzinger reaction; (4) The 2,4'-biquinoline-4-carboxylic acid obtained in step (3) is reacted with cyclopropylamine or N 1 ,N 1 -dimethyl-1,2-ethylenediamine condensation to prepare asymmetrically polyfluorinated 2,4'-biquinoline-4-amide derivatives.

4. The method for preparing an asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 3, characterized in that: In the step (1), the fluoroacetophenone is any one of o-fluoroacetophenone, m-fluoroacetophenone, p-fluoroacetophenone, 3-chloro-4-fluoro-acetophenone, 3,4-difluoroacetophenone, 2,3,4-trifluoroacetophenone and 2,3,4,5-tetrafluoroacetophenone; and the molar ratio of the fluoroacetophenone to 5-fluoroindigo red is 1:1 to 1.

5.

5. The method for preparing an asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 4, characterized in that: In the step (2), the molar ratio of the quinoline-4-carboxylic acid intermediate to the potassium salt of monoethyl malonate is 1:2-3.

6. The method for preparing an asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 5, characterized in that: In the step (3), the molar ratio of quinoline-4-ethanone to 5-fluoroindigo red is 1:1 to 1.

5.

7. The method for preparing an asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 6, characterized in that: In the step (4), 2,4'-biquinoline-4-carboxylic acid, a condensing agent and cyclopropylamine or N 1 ,N 1 -The molar ratio of dimethyl-1,2-ethylenediamine is 1:(1~3):(1~2).

8. The method for preparing an asymmetric polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 7, characterized in that: The condensation agent in step (4) is any one of thionyl chloride, phosphorus oxychloride, phosphorus pentoxide, polyphosphoric acid, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and carbonyldiimidazole.

9. Use of the asymmetrically polyfluoro-substituted 2,4'-biquinoline-4-amide derivative according to claim 1 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that: The anti-tumor drug is a drug for treating leukemia, liver cancer, pancreatic cancer or non-small cell lung cancer.

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