An N-substituted phenyl-pyridazinone-3-carboxamide compound, a preparation method and application thereof, and a pharmaceutical composition

Abstract

The present application relates to the technical field of pharmaceutical organic matter, especially to a kind of N-substituted phenyl-pyridazinone-3-carboxamide compound and its preparation method and application and a kind of pharmaceutical composition.The compound related to pneumonia described in the present application is N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compound, for treating sepsis, acute lung injury, and the preparation form that can be prepared includes injection, tablet, capsule, aerosol, suppository, film, drop pill, ointment, controlled release or sustained release agent and nano preparation.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical organics technology, and in particular to an N-substituted phenyl-pyridazinone-3-carboxamide compound, its preparation method and application, and a pharmaceutical composition thereof. Background Technology

[0002] Inflammation is a defensive response of the body to stimuli. Under normal circumstances, inflammation is beneficial and is an automatic defense mechanism of the body. However, when the inflammatory response is disordered and leads to excessive inflammation, the body produces a large number of inflammatory cytokines, which can cause damage to tissues or cells and seriously affect human health.

[0003] Sepsis is a systemic inflammatory response syndrome caused by the invasion of pathogenic microorganisms such as bacteria. In addition to the manifestations of systemic inflammatory response syndrome and primary infection focus, severely ill patients often exhibit organ insufficiency. The mortality rate of sepsis is as high as 30-70%. In recent years, despite significant progress in anti-infective therapy and organ function support techniques, there is still a lack of effective drugs for the clinical treatment of sepsis.

[0004] Clinical and animal studies have confirmed that lipopolysaccharide (LPS) can activate multiple downstream pro-inflammatory signaling pathways and trigger the excessive production of inflammatory factors, such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), leading to sepsis. Currently, the blocking of cytokine storms by various drugs, including glucocorticoids, has been extensively explored as a potential and promising method for the prevention and treatment of sepsis. However, given that clinically used drugs have not shown significant therapeutic effects on sepsis patients due to their low efficacy and adverse reactions, there is still a need to develop more effective and safer novel anti-inflammatory drugs to treat sepsis. Therefore, inhibiting the release of inflammatory factors such as TNF-α and IL-6 has become an important means of treating sepsis. Summary of the Invention

[0005] The purpose of this invention is to provide an N-substituted phenyl-pyridazinone-3-carboxamide compound, its preparation method and application, and a pharmaceutical composition to overcome the shortcomings of the prior art.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides an N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compound comprising one of the following structural formulas:

[0008]

[0009] The present invention also provides a method for preparing the N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compounds, comprising the following steps:

[0010] (1) Water, ethanol, ethyl acetoacetate and sodium acetate were mixed and then cooled to obtain system 1;

[0011] After adjusting the pH of the aqueous solution of aniline compounds to 1-2 and cooling it, it was then mixed with sodium nitrite solution to obtain system 2;

[0012] System 2 and System 1 were mixed and reacted to produce compound 2;

[0013] The aniline compounds include aniline, p-fluoroaniline, p-methylaniline, p-methoxyaniline, p-ethylaniline, or 2-ethylaniline;

[0014] (2) The obtained compound 2, solvent, ethoxycarbonylmethylenetriphenylphosphine and diethylamine were mixed and reacted to generate compound 3;

[0015] (3) The obtained compound 3, ethanol, 1,4-dioxane and alkaline solution were mixed and reacted to obtain a reaction solution;

[0016] The pH of the reaction solution was adjusted to 2-3 to generate compound 4;

[0017] (4) 4-chloro6,7-dimethoxyquinoline, 4-nitrophenol, potassium iodide, cesium carbonate and solvent were mixed and reacted to generate compound 7;

[0018] (5) After mixing compound 7, water, ethanol, reducing iron powder and ammonium chloride, the mixture was reacted to produce 4-[(6,7-dimethoxyquinoline-4-yl)oxy]aniline, which was designated as compound 8;

[0019] (6) Compound 8, compound 4, solvent, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and triethylamine were mixed and subjected to acid-amine condensation reaction to generate N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxamide compounds;

[0020] There is no requirement for the order of steps (1) to (3) to prepare compound 4 and steps (4) to (5) to prepare compound 8.

[0021] Preferably, in step (1), the ratio of water, ethanol, ethyl acetoacetate and sodium acetate is 1 mL: 2-6 mL: 2-3 mL: 1-2 g;

[0022] The independent cooling temperature is reduced to -2 to -6℃;

[0023] Adjust the pH value using concentrated hydrochloric acid;

[0024] The ratio of aniline compounds to sodium nitrite solution is 1.5–2 g: 15–25 mL, and the mass concentration of sodium nitrite solution is 45–55%.

[0025] The ratio of aniline compounds to ethyl acetoacetate is 1.5–2 g : 2–3 mL;

[0026] The volume ratio of aniline compounds to water in aqueous solutions is 1.5–2.5:1.

[0027] The reaction temperature is -5 to 5℃, and the time is 1 to 5 hours.

[0028] Preferably, the solvent in step (2) comprises dimethyl sulfoxide;

[0029] The ratio of aniline compounds, solvent, ethoxycarbonylmethylenetriphenylphosphine, and diethylamine is 1.5–2 g : 3–5 mL : 2–3 g : 0.05–0.1 mL;

[0030] The reaction temperature is 80–90℃, and the reaction time is 2–8 hours.

[0031] The mass concentration of the alkaline solution in step (3) is 5-15%;

[0032] The alkaline solution contains either sodium hydroxide solution or potassium hydroxide solution;

[0033] The ratio of aniline compounds, ethanol, 1,4-dioxane and alkaline solution is 1.5–2 g : 15–25 mL : 15–25 mL : 4–5 mL;

[0034] The reaction time is 0.5–2 hours;

[0035] Use concentrated hydrochloric acid to adjust the pH value.

[0036] Preferably, in step (4), the ratio of 4-chloro-6,7-dimethoxyquinoline, 4-nitrophenol, potassium iodide, cesium carbonate, and solvent is 0.2–0.8 mmol: 0.2–0.8 mmol: 0.02–0.08 mmol: 1–3 mmol: 3–8 mL;

[0037] The solvent is toluene;

[0038] The reaction is carried out at a temperature of 120–160°C for a time of 10–15 hours.

[0039] In step (5), the ratio of compound 7, water, ethanol, reduced iron powder and ammonium chloride is 0.5-1.5 mmol: 3-7 mL: 8-12 mL: 8-13 mmol: 1-3 mmol;

[0040] The reaction is carried out at a temperature of 40–80°C for 2–8 hours.

[0041] Preferably, in step (6), the ratio of compound 8, compound 4, solvent, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole, and triethylamine is 0.05–0.2 mmol: 0.1–0.3 mmol: 4–8 mL: 0.1–0.3 mmol: 0.1–0.3 mmol: 0.2–0.4 mmol;

[0042] The solvent is dichloromethane;

[0043] The reaction time is 10–20 hours.

[0044] The present invention also provides the use of the N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compounds in the preparation of anti-inflammatory drugs, which treat inflammation by inhibiting the release of inflammatory factors from macrophages, and are used to prevent or treat inflammation and inflammation-related diseases.

[0045] Preferably, the inflammation or inflammation-related disease includes, but is not limited to, sepsis, acute lung injury, arthritis, colitis, hepatitis, fatty liver, or chronic inflammatory disease.

[0046] The present invention also provides a pharmaceutical composition for the prevention or treatment of inflammation and inflammation-related diseases, comprising the N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compound or a pharmaceutically acceptable salt thereof, and further comprising pharmaceutical excipients.

[0047] Preferably, the pharmaceutical composition is formulated in the form of an injection, tablet, capsule, aerosol, suppository, film, drop, ointment, controlled-release or sustained-release formulation, and nanoformulation.

[0048] The N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compounds provided by this invention are used to treat sepsis and acute lung injury. The formulations that can be prepared include injections, tablets, capsules, aerosols, suppositories, films, pellets, ointments, controlled-release or sustained-release formulations, and nano-formulations. Attached Figure Description

[0049] Figure 1 The dose-response relationship of the compound inhibiting the release of IL-6 and TNF-α from LPS-stimulated macrophages;

[0050] Figure 2 The graph shows how the preferred compound (J27) improves the survival rate of septic mice and slows down the rate of weight loss in mice. Detailed Implementation

[0051] This invention provides a method for preparing the N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compounds, comprising the following steps:

[0052] (1) Water, ethanol, ethyl acetoacetate and sodium acetate were mixed and then cooled to obtain system 1;

[0053] After adjusting the pH of the aqueous solution of aniline compounds to 1-2 and cooling it, it was then mixed with sodium nitrite solution to obtain system 2;

[0054] System 2 and System 1 were mixed and reacted to produce compound 2;

[0055] The aniline compounds include aniline, p-fluoroaniline, p-methylaniline, p-methoxyaniline, p-ethylaniline, or 2-ethylaniline;

[0056] (2) The obtained compound 2, solvent, ethoxycarbonylmethylenetriphenylphosphine and diethylamine were mixed and reacted to generate compound 3;

[0057] (3) The obtained compound 3, ethanol, 1,4-dioxane and alkaline solution were mixed and reacted to obtain a reaction solution;

[0058] The pH of the reaction solution was adjusted to 2-3 to generate compound 4;

[0059] (4) 4-chloro6,7-dimethoxyquinoline, 4-nitrophenol, potassium iodide, cesium carbonate and solvent were mixed and reacted to generate compound 7;

[0060] (5) After mixing compound 7, water, ethanol, reducing iron powder and ammonium chloride, the mixture was reacted to produce 4-[(6,7-dimethoxyquinoline-4-yl)oxy]aniline, which was designated as compound 8;

[0061] (6) Compound 8, compound 4, solvent, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and triethylamine were mixed and subjected to acid-amine condensation reaction to generate N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxamide compounds;

[0062] There is no requirement for the order of steps (1) to (3) to prepare compound 4 and steps (4) to (5) to prepare compound 8.

[0063] In this invention, the ratio of water, ethanol, ethyl acetoacetate and sodium acetate in step (1) is 1 mL: 2-6 mL: 2-3 mL: 1-2 g, preferably 1 mL: 4-5 mL: 2.5-2.8 mL: 1.5-1.8 g;

[0064] The independent cooling is to a temperature of -2 to -6°C, preferably -4 to -5°C;

[0065] Adjust the pH value using concentrated hydrochloric acid;

[0066] The ratio of aniline compound to sodium nitrite solution is 1.5–2 g: 15–25 mL, preferably 1.6–1.8 g: 18–20 mL; the mass concentration of sodium nitrite solution is 45–55%, preferably 48–50%.

[0067] The ratio of aniline compounds to ethyl acetoacetate is 1.5–2 g : 2–3 mL, preferably 1.8–1.9 g : 2.3–2.5 mL;

[0068] The volume ratio of aniline compound to water in the aqueous solution is 1.5 to 2.5:1, preferably 2:1;

[0069] The reaction temperature is -5 to 5°C, preferably -4 to 4°C, more preferably -3 to 2°C, and even more preferably -1 to 0°C; the reaction time is 1 to 5 hours, preferably 2 to 3 hours.

[0070] In this invention, the solvent in step (2) comprises dimethyl sulfoxide;

[0071] The ratio of aniline compounds, solvent, ethoxycarbonylmethylenetriphenylphosphine, and diethylamine is 1.5–2 g: 3–5 mL: 2–3 g: 0.05–0.1 mL, preferably 1.6–1.8 g: 4 mL: 2.3–2.6 g: 0.06–0.08 mL;

[0072] The reaction temperature is 80–90°C, preferably 82–88°C, and more preferably 85–86°C; the reaction time is 2–8 hours, preferably 4–6 hours.

[0073] In step (3), the mass concentration of the alkaline solution is 5-15%, preferably 8-12%, and more preferably 9-10%; the alkaline solution contains sodium hydroxide solution or potassium hydroxide solution.

[0074] The ratio of aniline compounds, ethanol, 1,4-dioxane and alkaline solution is 1.5-2g:15-25mL:15-25mL:4-5mL, preferably 1.6-1.8g:18-20mL:18-20mL:4.5mL;

[0075] The reaction time is 0.5 to 2 hours, preferably 1 to 1.5 hours;

[0076] Use concentrated hydrochloric acid to adjust the pH value.

[0077] In this invention, the ratio of 4-chloro6,7-dimethoxyquinoline, 4-nitrophenol, potassium iodide, cesium carbonate and solvent in step (4) is 0.2-0.8 mmol: 0.2-0.8 mmol: 0.02-0.08 mmol: 1-3 mmol: 3-8 mL, preferably 0.4-0.6 mmol: 0.4-0.6 mmol: 0.04-0.05 mmol: 1.5-2 mmol: 5-6 mL;

[0078] The solvent is toluene;

[0079] The reaction temperature is 120–160°C, preferably 130–150°C, and more preferably 140–145°C; the reaction time is 10–15 h, preferably 12–13 h.

[0080] In step (5), the ratio of compound 7, water, ethanol, reduced iron powder and ammonium chloride is 0.5-1.5 mmol: 3-7 mL: 8-12 mL: 8-13 mmol: 1-3 mmol, preferably 0.8-1.2 mmol: 4-6 mL: 9-10 mL: 10-11 mmol: 2-2.3 mmol;

[0081] The reaction temperature is 40–80°C, preferably 50–70°C, and more preferably 55–60°C; the reaction time is 2–8 hours, preferably 4–6 hours.

[0082] In this invention, the ratio of compound 8, compound 4, solvent, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and triethylamine in step (6) is 0.05-0.2 mmol: 0.1-0.3 mmol: 4-8 mL: 0.1-0.3 mmol: 0.1-0.3 mmol: 0.2-0.4 mmol, preferably 0.1-0.15 mmol: 0.18-0.23 mmol: 6-7 mL: 0.15-0.2 mmol: 0.2-0.23 mmol: 0.3-0.34 mmol;

[0083] The solvent is dichloromethane;

[0084] The reaction time is 10 to 20 hours, preferably 14 to 17 hours.

[0085] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0086] The preparation routes of each embodiment of the present invention are as follows:

[0087]

[0088] Example 1

[0089] 4-Methyl-6-oxo-1-phenyl-1,6-dihydropyridazine-3-carboxylic acid (4a)

[0090] Step 1: In a 50 mL round-bottom flask, add water (1 mL), ethanol (4 mL), ethyl acetoacetate (2.31 mL, 0.0177 mol), and anhydrous sodium acetate (1.59 g, 0.0194 mol) sequentially. Mix thoroughly and stir at room temperature for 1 h. Then, lower the system temperature to -4 °C using a salt-ice bath. Next, dissolve aniline 1a (1.79 g, 0.019 mol) completely in deionized water (aniline to water volume ratio 2:1). Add concentrated hydrochloric acid to adjust the pH to 1-2, then transfer to a salt-ice bath to lower the temperature below -4 °C. Slowly add 20 mL of 50% sodium nitrite solution and stir at low temperature for 30 min to form system 2. After 30 min, add system 2 to system 1. During this process, a large amount of yellow solid will precipitate. Maintain the reaction at 0 °C for 2 h, then filter, wash, and dry to obtain 2a. The prepared 2a was dissolved in 4 mL of DMSO in a 250 mL round-bottom flask, followed by the addition of ethoxycarbonylmethylenetriphenylphosphine (CBMTPP) (2.23 g, 0.0064 mol) and diethylamine (Et₂NH) (0.0669 mL, 0.0006 mol). The reaction was carried out at 85 °C for 4 h. After the reaction was completed, the reactants were cooled to room temperature and poured into ice water. The mixture was extracted with dichloromethane, washed with saturated brine, and finally separated by column chromatography to obtain 3a. Finally, the obtained 3a was mixed with ethanol (20 mL) and 1,4-dioxane (20 mL) in a 100 mL round-bottom flask. Then, 5 mL of 10% NaOH solution was added to the mixture, and the mixture was stirred at room temperature for 1 h. After the reaction was completed, concentrated hydrochloric acid was added to the system to adjust the pH to 2-3. During this process, a pale yellow solid was precipitated. After filtration, washing and drying, 4-methyl-6-oxo-1-phenyl-1,6-dihydropyridazine-3-carboxylic acid 4a was obtained.

[0091] White power; yield: 50.1%. 1H NMR (400MHz, CDCl3) δ7.49(t,J=7.5Hz,2H),7.41(dd,J=15.4,7.6Hz,2H),7.37–7.28(m,1H),6.80(d,J=6.8Hz,1H),2.44(d,J=6.9Hz,3H).

[0092] Following the method in step one, 1-(4-fluorophenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid 4b was prepared from p-fluoroaniline as a raw material.

[0093] Yellowpower; yield: 56.5%. 1 H NMR (400MHz, CDCl3) δ7.53 (dd, J=8.0,

[0094] 4.9Hz,2H),7.11(t,J=8.2Hz,2H),6.81(s,1H),2.46(s,3H).

[0095] Following the method in step one, 4-methyl-6-oxo-1-(p-toluene)-1,6-dihydropyridazine-3-carboxylic acid 4c was prepared from p-methylaniline as a raw material.

[0096] Yellow power; yield: 52.3%. 1 H NMR (400MHz, MeOD) δ7.40(d,J=8.4Hz,2H),7.28(s,1H),7.26(s,1H),6.87(d,J=1.1Hz,1H),2.44(d,J=0.9Hz,3H),2.36(s,3H).

[0097] Following the method in step one, 1-(4-methoxyphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid 4d was prepared from p-methoxy group as a raw material.

[0098] Yellow power; yield: 59.8%. 1 H NMR (400MHz, CDCl3) δ7.01 (d, J=9.0

[0099] Hz, 2H), 6.49 (d, J = 9.0Hz, 2H), 6.37 (s, 1H), 3.35 (s, 3H), 2.03 (s, 3H).

[0100] Following the method in step one, 1-(4-ethylphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid 4e was prepared from p-ethylaniline as a raw material.

[0101] Yellow power; yield: 55.3%. 1 H NMR (400MHz, CDCl3) δ7.42 (d, J=8.3

[0102] Hz,2H),7.24(s,1H),6.82(s,1H),2.64(q,J=7.6Hz,2H),2.46(s,3H),1.19(t,J=7.6Hz,3H).

[0103] Following the method in step one, 1-(2-ylphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid 4f was prepared from 2-ethylaniline as a raw material.

[0104] Yellow power; yield: 58.1%. 1 H NMR (400MHz, CDCl3) δ7.37–7.29(m,2H),7.24(d,J=7.0Hz,1H),7.16(d,J=7.8H z,1H),6.82(s,1H),2.47(s,3H),2.38(q,J=7.5Hz,2H),1.07(t,J=7.6Hz,3H).

[0105] Step 2: In a 50 mL round-bottom flask, 4-chloro-6,7-dimethoxyquinoline 5 (100 mg, 0.448 mmol), 4-nitrophenol 6 (62.33 mg, 0.5 mmol), potassium iodide (7.44 mg, 0.045 mmol), and cesium carbonate (292.21 mg, 2 mmol) were dissolved in 5 mL of toluene solution and reacted at 140 °C for 12 h. When the reaction was complete as monitored by TLC, a pale yellow solid 7 was obtained by stirring with petroleum ether. Solid 7 (110 mg, 1 mmol) was dissolved in a mixed solution of water (5 mL) and ethanol (10 mL). Then, reduced iron powder (560 mg, 10 mmol) and ammonium chloride (130 mg, 2 mmol) were added to the solution, and the reaction was carried out at 65 °C for 4 h. After the reaction was complete as monitored by TLC, inorganic residues were removed by filtration, and the filtrate was dried under vacuum to obtain the crude product. Finally, column chromatography (DCM:CH3OH = 30:1) was used to separate the compound into 8, which is the target compound 4-((6,7-dimethoxyquinoline-4-yl)oxy)aniline.

[0106] White power; yield: 64.7%. 1H NMR (400MHz, MeOD) δ8.65(s,2H),8.34(d,J=8.3Hz,2H),8.05(dd,J=8.2,7.4Hz,2H),7.83–7.73(m,4H),4.92(s,3H),4.92(s,3H).

[0107] Step 3: Dissolve 4-((6,7-dimethoxyquinoline-4-yl)oxy)aniline (50 mg, 0.169 mmol) and 4-methyl-6-oxo-1-phenyl-1,6-dihydropyridazine-3-carboxylic acid (46.63 mg, 0.2 mmol) in ultra-dry dichloromethane (6 mL), add EDCI (38.84 mg, 0.2 mmol), HOBT (27.4 mg, 0.21 mmol) and TEA (36.81 mL, 0.3 mmol), and carry out the acid-amine condensation reaction at room temperature for 13 h. After the reaction was monitored by TLC, the dichloromethane was evaporated to dryness and then extracted with a DCM:MeOH (60:1) mixed solvent. After extraction, the mixture was dried with anhydrous magnesium sulfate, filtered, evaporated to dryness, and then separated by column chromatography (DCM:MeOH = 10:1) to obtain J25, namely N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-phenyl-1,6-dihydropyridazine-3-carboxamide.

[0108] N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-phenyl-1,6-dihydropyridazine-3-carboxamide (J25)

[0109] White solid; yielded: 69.3%; mp: 124.7–126.7℃. 1 H NMR (400MHz, CDCl3) δ9.02 (s, 1H), 8.46 (d, J = 5.0Hz, 1H), 7.71 (d, J = 8.9Hz, 2H), 7.60 (d, J = 7.8Hz, 2H), 7.54 (s, 1H), 7.51 (d, J = 8.0Hz, 1H), 7.4 6(t,J=7.3Hz,1H),7.41(s,1H),7.19(s,1H),7.17(s,1H),6.90(d,J=0. 9Hz, 1H), 6.44 (d, J = 5.3Hz, 1H), 4.03 (s, 3H), 3.98 (s, 3H), 2.66 (s, 3H). 13CNMR (100MHz, CDCl3) δ160.73,160.58,159.44,152.96,150.98,149.63,148.70,146.80,144.56,140.69,138.23,134.63,130. 92,129.11(×2),128.94,125.45(×2),122.04(×2),121.76(×2),116.08,107.76,103.36,99.48,56.18(×2),20.48.ESI-MS:m / z 509.2[M+H]+.

[0110] Following the method in step three, N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)aniline and 1-(4-fluorophenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid were prepared from 4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-1-(4-fluorophenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (J26).

[0111] White solid; yield 87.5%; mp:126.7-128.0℃; 1 H NMR(400MHz, CDCl3)δ9.04(s,1H),8.53(s,1H),7.78(s,2H),7.66(s,2H),7.60(s,1H),7.50(s,2H) ,7.28(s,1H),7.25–7.19(m,1H),6.97(s,1H),6.51(s,1H),4.11(s,3H),4.10(s,3H),2.73(s,3H). 13 C NMR (100MHz, CDCl3) δ163.59,160.78,160.46,159.38,152.99,150.97,149.63,148.58,146.66,144.72,138.43,136.59,134.58,130.90 ,128.85,127.42,127.34,122.05(×2),121.81(×2),116.17,115.94,107.60,103.30,99.44,56.19(×2),20.50.ESI-MS:m / z:527.2[M+H] + .

[0112] Following the method in step three, N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)aniline and 4-methyl-6-oxo-1-(p-toluene)-1,6-dihydropyridazine-3-carboxylic acid were prepared from 4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(p-toluene)-1,6-dihydropyridazine-3-carboxamide (J27).

[0113] White solid; yield 85.2%; mp:150.9-151.3℃; 1 H NMR (400MHz, CDCl3) δ9.03(s,1H),8.46(d,J=5.2Hz,1H),7.70(d,J=8.7Hz,2H),7.54(s,1H),7.47(d,J=8.0Hz,2H),7.40(s,1H),7. 32(d,J=7.9Hz,2H),7.18(d,J=8.6Hz,2H),6.88(s,1H),6.44(d,J=5.2Hz,1H),4.03(s,3H),3.97(s,3H),2.65(s,3H),2.42(s,3H). 13 C NMR (100MHz, CDCl3) δ160.70,160.63,159.55,152.90,150.93,149.58,148.80,146.87,144.47,139.17,138.24,137.99,134.65,130.83, 129.70(×2),125.26(×2),122.00(×2),121.78(×2),116.06,107.82,103.34,99.45,56.20,56.18,21.29,20.53.ESI-MS:m / z:523.2[M+H] + .

[0114] Following the method in step three, N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)aniline and 1-(4-methoxyphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid were prepared from 4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-1-(4-methoxyphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (J28).

[0115] White solid; yield 88.6%; mp:199.1-200.6℃; 1H NMR (400MHz, CDCl3) δ8.99(s,1H),8.47(d,J=5.3Hz,1H),7.70(d,J=8.8Hz,2H),7.57–7.49(m,3H),7.41(s,1H),7.19(d,J= 8.7Hz,2H),7.03(d,J=8.9Hz,2H),6.90(s,1H),6.45(d,J=5.2Hz,1H),4.04(s,3H),4.00(s,3H),3.87(s,3H),2.66(s,3H). 13 CNMR (100MHz, CDCl3) δ161.68,160.69,159.76,159.71,153.65,150.49,149.97,147.61,145.28,144.49,138.09,135.01,133.60,130.72 ,126.72(×2),122.09(×2),121.76(×2),116.05,114.24(×2),106.45,103.29,99.58,56.36,56.29,55.66,20.47.ESI-MS:m / z:539.2[M+H] + .

[0116] Following the method in step three, N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)aniline and 1-(4-ethylphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid were prepared from 4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-1-(4-ethylphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (J29).

[0117] White solid; yield 70.9%; mp:145.0-146.1℃; 1 H NMR(400MHz, CDCl3)δ8.98(s,1H),8.47(d,J=5.1Hz,1H),7.70(d,J=8.6Hz,2H),7.57–7.48(m,3H),7.41(s,1H),7.36(d,J=8.0Hz,2H),7.1 9(d,J=8.6Hz,2H),6.91(s,1H),6.45(d,J=5.1Hz,1H),4.09(s,3H),4.04(s,3H),2.74(q,J=7.5Hz,2H),2.67(s,3H),1.29(t,J=7.6Hz,3H). 13C NMR (100MHz, CDCl3) δ160.75,160.62,159.56,152.94,150.92,149.60,148.73,146.80,145.39,144.49,138.38,137.96,134.64,130.87,12 8.57(×2),125.32(×2),122.01(×2),121.79(×2),116.07,107.77,103 .34,99.46,56.21(×2),28.64,20.55,15.51.ESI-MS:m / z:537.2[M+H] + .

[0118] Following the method in step three, N-(4-((6,7-dimethoxyquinoline-4-yl)oxy)aniline and 1-(2-ylphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid were prepared from 4-((6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-1-(2-ethylphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (J30).

[0119] White solid; yield 81.3%; mp:248.9-250.7℃; 1 H NMR (400MHz, CDCl3) δ8.87(d,J=10.4Hz,1H),8.47(d,J=4.2Hz,1H),7.68(d,J=6.7Hz,2H),7.45(dd,J=35.4,21.3Hz,5H),7.30(s,1H) ,7.17(d,J=6.7Hz,2H),6.94(s,1H),6.45(s,1H),4.07(s,3H),4.04(s,3H),2.71(s,3H),2.52(d,J=6.3Hz,2H),1.20(d,J=2.1Hz,3H). 13 C NMR(100MHz,DMSO-d6)δ162.80,162.18,160.31,159.46,153.06,150.52 ,149.84,149.29,146.95,143.47,142.15,140.67,140.13,136.11,129.8 6,129.37,128.26,127.14,122.56(×2),121.88(×2),115.70,108.32,10 3.69,99.57,36.25,31.24,23.90,18.80,14.39.ESI-MS:m / z:537.2[M+H]+ .

[0120] Pharmacological studies of the product of this invention

[0121] Dose-response relationship of compounds in inhibiting the release of IL-6 and TNF-α from LPS-stimulated macrophages (J774A.1) in the examples.

[0122] When stimulated with LPS, J774A.1 cells secrete excessive amounts of pro-inflammatory cytokines (such as IL-6 and TNF-α). This invention establishes an enzyme-linked immunosorbent assay (ELISA) to test the anti-inflammatory activity of the compound against IL-6 and TNF-α release in LPS-stimulated J774A.1 cells. Dimethyl sulfoxide (DMSO) was used as a solvent control. J774A.1 macrophages were treated with 10 μM of the compound for 0.5 hours, followed by stimulation with LPS (0.5 μg / mL) and cultured for 24 hours. The levels of IL-6 and TNF-α were then determined using an ELISA kit. The cytokine-inhibiting activity of the compound is as follows: Figure 1 As shown, the results indicate that the six compounds can significantly inhibit the release of IL-6 and TNF-α from LPS-stimulated macrophages, exhibiting significant anti-inflammatory effects.

[0123] The physiological and pathological changes in septic mice were alleviated by the preferred compound J27.

[0124] This invention further explores the protective effect of the preferred compound J27 on a LPS-induced sepsis model in C57 / BL6 mice. Dimethyl sulfoxide was used as a solvent control. Mice were pretreated with either the preferred compound J27 (20 mg / kg, intraperitoneal injection) or the solvent, followed by stimulation with 15 mg / kg LPS. Figure 2 As shown, the preferred compound J27 can increase the survival rate of septic mice and alleviate the symptoms of weight loss in mice. As illustrated, after injection of a high dose of LPS to induce sepsis, all mice died within 36 hours, while half of the mice treated with J27 survived, and the treated mice gradually regained their weight after 48 hours. Furthermore, during the experiment, it was found that after 48 hours, the vital activities of the mice in the drug treatment group were not significantly different from those in the negative control group. This indicates that the compound can effectively alleviate the physiological changes in septic mice.

[0125] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A compound of the formula N-(4-(6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-6-oxo- 1,6-dihydropyridazine-3-carboxamide, characterized by, comprising one of the following structural formulae:

2. The method for preparing the N-(4-(6,7-dimethoxyquinoline-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compound according to claim 1, characterized in that, comprising the following steps: (1) mixing water, ethanol, ethyl acetoacetate and sodium acetate and then cooling to obtain system 1; adjusting the pH value of aniline compound 1a-1f aqueous solution to 1-2 and then cooling, and then mixing with sodium nitrite solution to obtain system 2; mixing system 2 and system 1 to react to generate compound 2a-2f; (2) mixing the obtained compound 2a-2f, solvent, ethoxycarbonylmethylidene triphenyl phosphonium and diethylamine to react to generate compound 3a-3f; (3) mixing the obtained compound 3a-3f, ethanol, 1,4-dioxane and lye to react to obtain a reaction solution; adjusting the pH value of the reaction solution to 2-3 to generate compound 4a-4f; (4) mixing 4-chloro-6,7-dimethoxyquinoline, 4-nitrophenol, potassium iodide, cesium carbonate and solvent to react to generate compound 7; (5) mixing compound 7, water, ethanol, reducing iron powder and ammonium chloride to react to generate 4-[(6,7-dimethoxyquinolin-4-yl)oxy]aniline, denoted as compound 8; (6) mixing compound 8, compound 4a-4f, solvent, 1-ethyl-(3-dimethylaminopropyl) carbonyl diimide hydrochloride, 1-hydroxybenzotriazole and triethylamine to perform acid amine condensation reaction to generate N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxamide compound; steps (1)-(3) for preparing compound 4a-4f and steps (4)-(5) for preparing compound 8 are not limited in sequence; the preparation route of the N-(4-(6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-6-oxo-1,6-dihydrodiazine-3-carboxamide compound is shown as follows:

3. The production method according to claim 2, characterized by, in step (1), the amount ratio of water, ethanol, ethyl acetoacetate and sodium acetate is 1 mL: 2-6 mL: 2-3 mL: 1-2 g; the cooling is independently cooled to -2 to -6℃; concentrated hydrochloric acid is used to adjust the pH value; the amount ratio of aniline compound 1a-1f and sodium nitrite solution is 1.5-2 g: 15-25 mL, and the mass concentration of sodium nitrite solution is 45-55%; the amount ratio of aniline compound 1a-1f and ethyl acetoacetate is 1.5-2 g: 2-3 mL; the volume ratio of aniline compound 1a-1f and water in aniline compound 1a-1f aqueous solution is 1.5-2.5: 1; the reaction temperature is -5 to 5℃, and the reaction time is 1-5 h.

4. The production method according to claim 2 or 3, characterized by, in step (2), the solvent is selected from dimethyl sulfoxide; the amount ratio of aniline compound 1a-1f, solvent, ethoxycarbonylmethylidene triphenyl phosphonium and diethylamine is 1.5-2 g: 3-5 mL: 2-3 g: 0.05-0.1 mL; the reaction temperature is 80-90℃, and the reaction time is 2-8 h; in step (3), the mass concentration of lye is 5-15%; the lye is selected from sodium hydroxide solution or potassium hydroxide solution; The aniline compound 1a-1f, ethanol, 1,4-dioxane and alkali solution are used in a ratio of 1.5-2 g:15-25 mL:15-25 mL:4-5 mL; The reaction time is 0.5-2 h; The pH value is adjusted by using concentrated hydrochloric acid.

5. The production method according to claim 4, characterized by, The 4-chloro-6,7-dimethoxyquinoline, 4-nitrophenol, potassium iodide, cesium carbonate and solvent used in the step (4) are in a ratio of 0.2-0.8 mmol:0.2-0.8 mmol:0.02-0.08 mmol:1-3 mmol:3-8 mL; The solvent is toluene; The reaction temperature is 120-160℃ and the reaction time is 10-15 h; The compound 7, water, ethanol, reducing iron powder and ammonium chloride used in the step (5) are in a ratio of 0.5-1.5 mmol:3-7 mL:8-12 mL:8-13 mmol:1-3 mmol; The reaction temperature is 40-80℃ and the reaction time is 2-8 h.

6. The production method according to claim 2 or 3 or 5, characterized by, The compound 8, compound 4a-4f, solvent, 1-ethyl-(3-dimethylaminopropyl) carbonyl diimide hydrochloride, 1-hydroxybenzotriazole and triethylamine used in the step (6) are in a ratio of 0.05-0.2 mmol:0.1-0.3 mmol:4-8 mL:0.1-0.3 mmol:0.1-0.3 mmol:0.2-0.4 mmol; The solvent is dichloromethane; The reaction time is 10-20 h.

7. The use of the N-(4-(6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compound in claim 1 in the preparation of an anti-inflammatory drug for treating inflammation by inhibiting the release of inflammatory factors released by macrophages, the anti-inflammatory drug being used for preventing or treating inflammation and diseases related to inflammation.

8. Use according to claim 7, characterized in that, The inflammation or the disease related to inflammation is selected from sepsis, acute lung injury, arthritis, colitis, hepatitis, fatty liver or chronic inflammatory disease.

9. A pharmaceutical composition for preventing or treating inflammation and diseases associated with inflammation, characterized by, The pharmaceutical composition further comprises a pharmaceutically acceptable salt of the N-(4-(6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide compound in claim 1.

10. The pharmaceutical composition of claim 9, wherein, The preparation form of the pharmaceutical composition is selected from injection, tablet, capsule, aerosol, suppository, film, drop pill, ointment, controlled release or sustained release preparation and nano preparation. The preparation form of the pharmaceutical composition is selected from injection, tablet, capsule, aerosol, suppository, film, drop pill, ointment, controlled release or sustained release preparation and nano preparation.

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