1-aryl-5-pyrazolone compound as well as pharmaceutical composition and application thereof

Abstract

The invention discloses a 1-aryl-5-pyrazolone compound as well as a pharmaceutical composition and application of the 1-aryl-5-pyrazolone compound. The compound has a structure selected from any one of the following structures, and also comprises a tautomer, a pharmaceutically acceptable salt or a mixture of the tautomer and the pharmaceutically acceptable salt: # imgabs0, which can effectively inhibit COX-2 and effectively scavenge free radicals at a micromolar concentration level, and can reduce side effects of heart and cerebral vessels and digestive tracts caused by NSAID induced free radicals, so that the compound can be used for treating cardiovascular and cerebrovascular diseases. And through the synergistic effect of anti-oxidative stress, anti-inflammatory and analgesic effects of the traditional Chinese medicine composition, a better treatment effect on chronic inflammatory diseases accompanied by pains can be achieved.

Description

Technical Field

[0001] The present invention relates to a 1-aryl-5-pyrazolone compound, its pharmaceutical composition and application, and particularly relates to a 1-aryl-5-pyrazolone compound having COX-2 inhibitory activity and free radical scavenging activity, its pharmaceutical composition and application. Background Art

[0002] Non-steroidal anti-inflammatory drugs (NSAIDs) are one of the most commonly used drugs in clinical practice and can be used for anti-inflammatory, analgesic and antipyretic effects in a variety of clinical diseases. NSAIDs include non-selective NSAIDs and COX-2 selective NSAIDs / COX-2 selective inhibitors, and the latter have lower gastrointestinal side effects than non-selective NSAIDs. Therefore, compared with non-selective NSAIDs, COX-2 selective inhibitors are more suitable for the treatment of chronic inflammatory diseases with pain that require long-term medication, such as rheumatoid arthritis, osteoarthritis, psoriatic arthritis and ankylosing spondylitis.

[0003] Although COX-2 selective inhibition solves the problem of gastrointestinal side effects caused by non-selective inhibitors, it increases the risk of cardiovascular events and stroke. However, it has been reported that NSAIDs can induce the production of reactive oxygen species (ROS) in different cells such as cardiac, cardiovascular, gastrointestinal cells, etc. Therefore, it is considered that the free radical production induced by NSAIDs is one of the mechanisms leading to its cardiovascular and gastrointestinal side effects. Summary of the Invention

[0004] Object of the Invention: The first object of the present invention is to provide a 1-aryl-5-pyrazolone compound, the second object is to provide a pharmaceutical composition containing the compound, and the third object is to provide an application of the compound and its pharmaceutical composition.

[0005] Technical Solution: The 1-aryl-5-pyrazolone compound described in the present invention has a structure selected from any of the following, and also includes its tautomer, pharmaceutically acceptable salt or a mixture thereof:

[0006]

[0007] Wherein:

[0008] R 1 is selected from C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, 3-6 membered cycloalkyl, 3-6 membered halocycloalkyl;

[0009] R 2 is selected from hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy;

[0010] R 3 is selected from hydrogen, halogen;

[0011] R 4 and R 5 are each independently selected from hydrogen, -(CH 2 ) n -SO 2 -(C1-C4 alkyl), -(CH 2 ) n -CO-(C1-C4 alkyl), -(CH 2 ) n -COOH, -(CH 2 ) n -CO-NH-OH;

[0012] n is an integer selected from 0 to 3.

[0013] Preferably, in the structure:

[0014] R 1 is selected from C1-C4 alkyl, C1-C4 haloalkyl, 3-5 membered cycloalkyl, 3-5 membered halocycloalkyl;

[0015] R 2 is selected from hydrogen, halogen, C1-C4 alkoxy, C1-C4 haloalkoxy;

[0016] R 3 is selected from hydrogen, halogen;

[0017] R 4 and R 5 are each independently selected from hydrogen, -(CH 2 ) n -SO 2 -(C1-C4 alkyl), -(CH 2 ) n -CO-(C1-C4 alkyl), -(CH 2 ) n -COOH, -(CH 2 ) n -CO-NH-OH;

[0018] n is selected from 0, 1.

[0019] More preferably, in the structure:

[0020] R 1 is selected from C1-C4 alkyl, 3-5 membered cycloalkyl;

[0021] R 2 is selected from hydrogen, halogen, C1-C4 alkoxy;

[0022] R 3Selected from hydrogen, halogen;

[0023] R 4 、R 5 Selected from hydrogen, -SO 2 -(C1-C4 alkyl), -(CH 2 ) n -CO-(C1-C4 alkyl), -(CH 2 ) n -COOH, -CO-NH-OH;

[0024] n is selected from 0, 1.

[0025] More preferably, in the structure:

[0026] R 1 Selected from methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl;

[0027] R 2 Selected from hydrogen, fluorine, chlorine, methoxy, ethoxy;

[0028] R 3 Selected from hydrogen, fluorine, chlorine;

[0029] R 4 、R 5 Selected from hydrogen, -SO 2 -CH 3 、-SO 2 -CH 2 CH 3 、-(CH 2 ) n -CO-CH 3 、-(CH 2 ) n -CO-CH 2 CH 3 、-(CH 2 ) n -COOH, -CO-NH-OH;

[0030] n is selected from 0, 1.

[0031] Even more preferably, in the structure:

[0032] R 2 、R 3 When they are both disubstituted, they are both hydrogen or chlorine.

[0033] R 4 、R 5 Are not both hydrogen, and R 4 、R 5 Are different;

[0034] Most preferably, the 1-aryl-5-pyrazolone compounds of the present invention are selected from any one of the following compounds:

[0035]

[0036]

[0037] Wherein, the pharmaceutically acceptable salt is a salt formed by the compound and an acid or a base, and the acid is 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, ferulic acid, and the base is an inorganic base containing an alkaline metal cation, an alkaline earth metal cation or an ammonium cation salt.

[0038] The chemical name of edaravone is 3-methyl-1-aryl-5-pyrazolone, and it is a free radical scavenger. Currently, edaravone is used clinically as a neuroprotective agent mainly for the treatment of stroke. Since oxidative stress caused by free radicals is an important factor in tissue damage caused by various diseases, edaravone not only has a neuroprotective effect clinically, but also shows good protective effects in animal models of tissue damage caused by various diseases. In addition, since oxidative stress caused by free radicals is also an important mechanism for the occurrence and development of cardiovascular diseases and arthritis diseases, current free radical scavengers have application potential for cardiovascular diseases and arthritis diseases. Edaravone shows certain therapeutic effects in animal models of various cardiovascular diseases and arthritis diseases.

[0039] The 1-aryl-5-pyrazolone compounds designed by the present invention are bifunctional molecules obtained by molecular hybridization of edaravone and a COX-2 selective inhibitor. It simultaneously has the effects of scavenging free radicals and selectively inhibiting COX-2, can reduce the cardiovascular and cerebrovascular and gastrointestinal side effects caused by NSAID-induced free radical generation, and can also exert better therapeutic effects on chronic inflammatory diseases accompanied by pain through its synergistic pharmacological effects of antioxidant stress and anti-inflammatory analgesia.

[0040] The preparation method of the 1-aryl-5-pyrazolone compounds of the present invention is as follows:

[0041]

[0042] Using the corresponding R 1 、R 2 and R 3Using substituted edaravone analogs as raw materials, through aldol condensation reaction, reduction reaction, or phenylsulfone series compounds and phenyl fatty acid ester series compounds, and then using benzoate series compounds as intermediates, respectively obtaining phenyl fatty acid series compounds and phenyl isohydroxamic acid series compounds through hydrolysis reaction and isohydroxamic acidification reaction.

[0043] Among them, R 1 , R 2 , R 3 , R 4 , R 5 are defined as described above;

[0044] Salifying the corresponding acid or base with the 1-aryl-5-pyrazolone compounds prepared by the above method, the pharmaceutically acceptable salts of the said compounds can be obtained.

[0045] "Pharmaceutically acceptable salts" refer to salts of compounds, which are prepared from compounds with specific substituents and relatively non-toxic acids or bases. When a compound contains relatively acidic functional groups, base addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When a compound contains relatively basic functional groups, acid addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, and the inorganic acids include, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid (forming carbonates or bicarbonates), phosphoric acid (forming phosphates, monohydrogen phosphates, dihydrogen phosphates), sulfuric acid (forming sulfates or bisulfates), hydroiodic acid, phosphorous acid, etc.; and organic acid salts, and the organic acids include, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid and similar acids; organic acid salts also include salts of amino acids (such as arginine, etc.), glucuronic acid and other organic acids. When certain specific compounds contain both basic and acidic functional groups, they can thus be converted into either base or acid addition salts. Preferably, the salt is contacted with a base or an acid in a conventional manner, and then the parent compound is separated, thereby regenerating the free form of the compound. The free form of the compound differs from its various salt forms in certain physical properties, such as different solubilities in polar solvents.

[0046] "Pharmaceutically acceptable salts" can be synthesized from parent compounds containing acid or base groups by conventional chemical methods. In general, the preparation method of such salts is to react these compounds in the form of free acids or bases with a stoichiometric amount of appropriate bases or acids in water or organic solvents or a mixture of both. Generally, non-aqueous media such as ethers, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.

[0047] The pharmaceutical composition described in the present invention comprises the 1-aryl-5-pyrazolone compounds and a pharmaceutically acceptable carrier.

[0048] The pharmaceutical composition also contains a pharmaceutically acceptable carrier.

[0049] "Pharmaceutically acceptable carrier" can be excipients widely used in the field of drug production. Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide methods to enable the active ingredient to dissolve at the desired rate after the subject receives the administration, or to promote the effective absorption of the active ingredient after the subject receives the composition administration. The described pharmaceutical excipients can be inert fillers or provide certain functions, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The described pharmaceutical excipients can include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesives, glidants, wetting agents, gelling agents, absorption retardants, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

[0050] The pharmaceutical composition described in the present invention can be prepared by any method known to those skilled in the art according to the disclosed content. For example, conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or freeze-drying processes.

[0051] The pharmaceutical composition of the present invention can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intra - arterial, intramuscular) administration. The pharmaceutical composition of the present invention can also be in a controlled - release or sustained - release dosage form (such as liposomes or microspheres). Examples of solid oral preparations include, but are not limited to, powders, capsules, caplets, soft gelatin capsules and tablets. Examples of liquid preparations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs and solutions. Examples of topical preparations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serum preparations. Examples of preparations for parenteral administration include, but are not limited to, injectable solutions, dry powder preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, injectable suspensions and injectable emulsions. Examples of other suitable preparations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols, such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

[0052] The 1 - aryl - 5 - pyrazolone compounds of the present invention are used in the preparation of COX - 2 inhibitors and free - radical scavenger drugs.

[0053] Preferably, the drug is an anti - inflammatory, anti - oxidative stress and analgesic drug.

[0054] More preferably, the drug is a drug for preventing and / or treating immune and inflammatory diseases related to the PGE - 2 signaling pathway, specifically involving inflammatory diseases with pain such as psoriatic arthritis, rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, etc.

[0055] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages:

[0056] These compounds can effectively inhibit COX - 2 and effectively scavenge free radicals at the micromolar concentration level, can reduce the cardiovascular and cerebrovascular and gastrointestinal side effects caused by NSAID - induced free - radical production, and may also exert a better therapeutic effect on chronic inflammatory diseases with pain through their synergistic effects of anti - oxidative stress, anti - inflammation and analgesia. Detailed implementation mode

[0057] The technical solutions of the present invention will be further described below in conjunction with examples.

[0058] Example 1

[0059]

[0060] Using an acetic acid solution of 1-phenyl-3-alkyl-5-pyrazolone raw material (2 mmol) and p-formylbenzenesulfone raw material (2.1 mmol), under room temperature conditions, magnetically stir for 12 hours until the reaction is complete. Filter the reaction solution to obtain a red precipitate, elute the residual acetic acid with petroleum ether, and remove the residual organic solvent by evaporation under reduced pressure to obtain the following intermediate. A dioxane solution of 0.5 mmol intermediate is slowly dropped into a 50% dioxane aqueous solution (V / V) containing 2 mmol sodium borohydride at room temperature. After the color of the reaction solution completely fades, adjust the pH of the reaction solution to neutral with dilute hydrochloric acid, then extract three times with ethyl acetate, combine the organic phases, remove water with anhydrous sodium sulfate, concentrate under reduced pressure, and separate by silica gel column chromatography with a petroleum ether / ethyl acetate eluent to obtain the final product. 1 HNMR(300MHz,DMSO)δ10.94(s,1H),7.84(d,J=7.9Hz,2H),7.73(d,J=8.0Hz,2H),7.62 - 7.29(m,4H),7.20(s,1H),3.75(d,2H),3.17(s,3H),2.09(d,3H).

[0061] Example 2

[0062]

[0063] Prepared according to the method of Example 1, using 1-phenyl-3-alkyl-5-pyrazolone raw material (2 mmol) and m-formylbenzenesulfone raw material (2.1 mmol). 1 HNMR(400MHz,DMSO)δ10.94(s,1H),7.88 - 7.68(m,4H),7.58(q,J=8.5,6.2Hz,2H),7.44(dd,J=8.6,7.3Hz,2H),7.20(s,1H),3.76(d,2H),3.20(s,3H),2.08(d,3H).

[0064] Example 3

[0065]

[0066] Prepared according to the method of Example 1, using 1-phenyl-3-alkyl-5-pyrazolone raw material and methyl p-formylphenylacetate raw material. 1 H NMR(300MHz,DMSO)δ10.80(s,1H),7.74(d,J=8.1Hz,2H),7.43(t,J=7.3Hz,2H),7.36 - 6.83(m,5H),3.62(s,4H),3.60(d,J=1.2Hz,4H),2.07(d,3H).

[0067] Example 4

[0068]

[0069] Prepared according to the method of Example 1 using 1-phenyl-3-alkyl-5-pyrazolone raw material and methyl p-formylbenzoate raw material. 1 H NMR(400MHz,DMSO)δ10.90(s,1H),7.88(d,J=8.3Hz,2H),7.74(dd,J=8.7,1.2Hz,2H),7.46-7.40(m,2H),7.37(d,J=8.0Hz,2H),7.19(t,J=7.4Hz,1H),3.83(s,3H),3.71(s,2H),2.06(s,3H).

[0070] Example 5

[0071]

[0072] Prepared according to the method of Example 1 using 1-phenyl-3-alkyl-5-pyrazolone raw material and methyl m-formylphenylacetate raw material. 1 HNMR(300MHz,DMSO)δ10.91(s,1H),7.89-7.65(m,4H),7.61-7.35(m,4H),7.19(t,J=7.4Hz,1H),3.84(s,3H),3.70(s,2H),2.07(s,3H).

[0073] Example 6

[0074]

[0075] Prepared according to the method of Example 1 using 3-ethyl-1-phenyl-1H-pyrazol-5-ol and methyl p-formylbenzoate raw materials. 1 H NMR(300MHz,DMSO)δ10.89(s,1H),7.88(d,J=8.3Hz,2H),7.75(dd,J=8.7,1.2Hz,2H),7.44(dd,J=8.6,7.3Hz,2H),7.37(d,J=7.9Hz,2H),7.20(t,J=7.4Hz,1H),3.83(s,3H),3.73(s,2H),2.46(s,2H),1.04(t,3H).

[0076] Example 7

[0077]

[0078] Prepared according to the method of Example 1, using 3-cyclopropyl-1-phenyl-1H-pyrazol-5-ol and methyl 4-formylbenzoate as raw materials. 1 HNMR(300MHz,DMSO)δ10.58(d,1H),7.89(d,J=7.9Hz,2H),7.69(d,J=7.4Hz,2H),7.47-7.35(m,4H),7.19(s,1H),3.91(s,1H),3.83(s,3H),3.71(s,1H),1.80(d,1H),0.96(s,2H),0.69(s,2H).

[0079] Example 8

[0080]

[0081] Prepared according to the method of Example 1, using 3-tert-butyl-1-phenyl-1H-pyrazol-5-ol and methyl 4-formylbenzoate as raw materials. 1 HNMR(300MHz,DMSO)δ10.55(d,1H),7.89(d,J=7.9Hz,2H),7.77(t,J=8.7Hz,2H),7.46(t,J=7.7Hz,2H),7.35-7.19(m,3H),3.91(d,J=51.9Hz,5H),1.19(d,9H).

[0082] Example 9

[0083]

[0084] Prepared according to the method of Example 1, using 3-methyl-1-(3-chlorophenyl)-1H-pyrazol-5-ol and methyl 4-formylbenzoate as raw materials. 1 HNMR(300MHz,DMSO)δ11.13(d,1H),7.88(d,J=8.1Hz,3H),7.73(d,J=7.9Hz,1H),7.47(t,J=8.2Hz,1H),7.32(d,3H),3.83(s,5H),2.06(d,3H).

[0085] Example 10

[0086]

[0087] Prepared according to the method of Example 1, using 3-methyl-1-(4-chlorophenyl)-1H-pyrazol-5-ol and methyl 4-formylbenzoate as raw materials. 11H NMR (300 MHz, DMSO) δ 11.11 (d, 1H), 7.88 (d, J = 8.2 Hz, 2H), 7.79 (d, J = 8.9 Hz, 2H), 7.49 (d, J = 8.9 Hz, 2H), 7.37 (d, J = 7.9 Hz, 2H), 3.83 (m, 5H), 2.06 (d, 3H).

[0088] Example 11

[0089]

[0090] Prepared according to the method of Example 1 using 1-(3-methoxyphenyl)-3-methyl-1H-pyrazol-5-ol and methyl 4-formylbenzoate as raw materials. 1 1H NMR (300 MHz, DMSO) δ 10.99 (s, 1H), 7.88 (d, J = 8.3 Hz, 2H), 7.58 - 7.18 (m, 5H), 6.76 (dt, J = 5.5, 2.7 Hz, 1H), 3.83 (s, 3H), 3.78 (s, 3H), 3.69 (s, 2H), 2.06 (s, 3H).

[0091] Example 12

[0092]

[0093] Prepared according to the method of Example 1 using 1-(3,4-dichlorophenyl)-3-methyl-1H-pyrazol-5-ol and methyl 4-formylbenzoate as raw materials. 1 1H NMR (300 MHz, DMSO) δ 11.12 (s, 1H), 8.06 (s, 1H), 7.88 (d, J = 8.2 Hz, 2H), 7.79 (dd, J = 8.9, 2.5 Hz, 1H), 7.70 (d, J = 8.9 Hz, 1H), 7.37 (d, J = 7.9 Hz, 2H), 3.83 (s, 3H), 3.70 (s, 2H), 2.08 (s, 3H).

[0094] Example 13

[0095]

[0096] Prepared using the end product of Example 3 as the raw material: 0.9 mmol of aqueous sodium hydroxide solution was added to a methanol solution of 0.3 mmol of the raw material, and the mixture was magnetically stirred at 50 °C for 12 hours until the reaction was complete. The reaction solution was naturally cooled to room temperature, quenched with dilute hydrochloric acid aqueous solution, concentrated under reduced pressure to precipitate the target product, filtered to obtain the crude product, dissolved in methanol, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and recrystallized to obtain the end product. 1¹H NMR (400 MHz, DMSO) δ 12.28 (s, 1H), 10.76 (s, 1H), 7.74 (d, J = 8.1 Hz, 2H), 7.48 - 7.39 (m, 2H), 7.16 (d, J = 10.9 Hz, 5H), 3.50 d, 4H), 2.05 (d, 3H)..

[0097] Example 14

[0098]

[0099] Prepared according to the method of Example 13, using the end product of Example 4 as the raw material. 1 ¹H NMR (300 MHz, DMSO) δ 12.83 (s, 1H), 10.89 (s, 1H), 7.86 (d, J = 7.9 Hz, 2H), 7.74 (d, J = 8.0 Hz, 2H), 7.43 (t, J = 7.9 Hz, 2H), 7.27 (d, J = 45.1 Hz, 3H), 3.71 (d, 2H), 2.05 (d, 3H).

[0100] Example 15

[0101]

[0102] Prepared according to the method of Example 13, using the end product of Example 5 as the raw material. 1 ¹H NMR (300 MHz, DMSO) δ 7.76 (m, 4H), 7.52 - 7.31 (m, 4H), 7.15 (t, J = 7.4 Hz, 1H), 3.69 (s, 2H), 2.03 (s, 3H).

[0103] Example 16

[0104]

[0105] Prepared according to the method of Example 13, using the end product of Example 6 as the raw material. 1 ¹H NMR (400 MHz, DMSO) δ 12.81 (s, 1H), 10.85 (d, 1H), 7.86 (d, J = 7.7 Hz, 2H), 7.75 (d, J = 8.1 Hz, 2H), 7.44 (t, J = 7.9 Hz, 2H), 7.34 (m, 2H), 7.21 (p, J = 7.7, 7.2 Hz, 1H), 3.72 (d, 2H), 2.53 (d, 1H), 2.34 (s, 1H), 1.06 (dt, 3H).

[0106] Example 17

[0107]

[0108] Prepared by using the end product of Example 7 as the raw material according to the method of Example 13. 1 H NMR(400MHz,DMSO)δ12.81(s,1H),10.55(d,1H),7.87(t,J=5.9Hz,2H),7.70(d,J=8.1Hz,2H),7.47-7.29(m,4H),7.19(m,1H),3.80(d,2H),1.81(dt,1H),0.83(dd,4H).

[0109] Example 18

[0110]

[0111] Prepared by using the end product of Example 8 as the raw material according to the method of Example 13. 1 HNMR(400MHz,DMSO)δ12.80(s,1H),10.53(d,1H),7.86(d,J=8.0Hz,2H),7.77(d,J=8.0Hz,2H),7.46(t,J=7.8Hz,2H),7.34-7.13(m,3H),3.90(d,2H),1.20(d,9H).

[0112] Example 19

[0113]

[0114] Prepared by using the end product of Example 9 as the raw material according to the method of Example 13. 1 HNMR(300MHz,DMSO)δ12.13(s,1H),7.95(q,J=2.1,1.6Hz,1H),7.88-7.72(m,3H),7.40(td,J=8.2,1.0Hz,1H),7.32(d,J=8.1Hz,2H),7.18(ddt,J=8.0,2.2,1.0Hz,1H),3.69(s,2H),2.03(s,3H).

[0115] Example 20

[0116]

[0117] Prepared by using the end product of Example 10 as the raw material according to the method of Example 13. 11H NMR (300 MHz, DMSO) δ 12.82 (s, 1H), 11.30 (s, 1H), 7.90 - 7.78 (m, 4H), 7.52 - 7.44 (m, 2H), 7.34 (d, J = 8.0 Hz, 2H), 3.71 (s, 2H), 2.06 (s, 3H).

[0118] Example 21

[0119]

[0120] Prepared according to the method of Example 13 using the end product of Example 11 as the raw material. 1 1H NMR (400 MHz, DMSO) δ 12.84 (s, 1H), 7.86 (d, J = 8.0 Hz, 2H), 7.53 - 7.18 (m, 5H), 6.78 (dq, J = 6.9, 3.7, 3.1 Hz, 1H), 3.78 (s, 3H), 3.68 (s, 2H), 2.07 (s, 3H).

[0121] Example 22

[0122]

[0123] Prepared according to the method of Example 13 using the end product of Example 12 as the raw material. 1 1H NMR (300 MHz, DMSO) δ 12.83 (s, 1H), 11.20 (s, 1H), 8.13 - 8.00 (m, 1H), 7.86 (d, J = 8.2 Hz, 2H), 7.80 (dd, J = 8.9, 2.5 Hz, 1H), 7.70 (d, J = 8.9 Hz, 1H), 7.34 (d, J = 8.1 Hz, 2H), 3.69 (s, 2H), 2.08 (s, 3H).

[0124] Example 23

[0125]

[0126] Prepared using the end product of Example 4 as the raw material: Hydroxylamine hydrochloride (1 equiv) and potassium hydroxide (1.1 equiv) were dissolved in methanol respectively, then mixed in an ice bath. After complete precipitation of potassium chloride, the newly prepared hydroxylamine methanol solution was obtained by filtration and stored at low temperature. 0.6 mmol of the raw material was dissolved in methanol, 6 mmol of the hydroxylamine methanol solution was added, and the hydrolysis reaction was carried out at room temperature for 12 h. The reaction solution was extracted with ethyl acetate / water, the organic layers were combined, dried over anhydrous Na 2 SO 4 , filtered, concentrated under reduced pressure, and finally the product was obtained by silica gel column chromatography. 1HNMR(300MHz, DMSO) δ 11.13 (s, 1H), 8.98 (s, 1H), 7.57 (t, J = 8.0 Hz, 4H), 7.36 (t, J = 7.9 Hz, 2H), 7.22 (d, J = 8.0 Hz, 2H), 7.15 (t, J = 7.4 Hz, 1H), 6.79 (s, 1H), 3.15 (d, J = 3.4 Hz, 2H), 2.13 (s, 3H).

[0127] Example 24

[0128]

[0129] Prepared using the end product of Example 5 as the raw material: According to the method of Example 23. 1 HNMR(300MHz, DMSO) δ 11.16 (s, 1H), 9.02 (s, 1H), 7.65 - 7.48 (m, 4H), 7.34 (t, J = 7.8 Hz, 2H), 7.26 (d, J = 5.0 Hz, 2H), 7.14 (t, J = 7.3 Hz, 1H), 6.78 (s, 1H), 3.14 (d, J = 2.6 Hz, 2H), 2.12 (s, 3H).

[0130] Example 25

[0131]

[0132] Prepared using the end product of Example 6 as the raw material: According to the method of Example 23. 1 HNMR(300MHz, DMSO) δ 11.14 (s, 1H), 10.83 (s, 1H), 8.98 (s, 1H), 7.78 - 7.72 (m, 2H), 7.66 (d, J = 8.2 Hz, 2H), 7.44 (dd, J = 8.6, 7.3 Hz, 2H), 7.29 (d, J = 7.9 Hz, 2H), 7.20 (t, J = 7.3 Hz, 1H), 3.68 (s, 2H), 2.50 (s, 2H), 1.07 (s, 3H).

[0133] Example 26

[0134]

[0135] Prepared using the end product of Example 7 as the raw material: According to the method of Example 23. 11H NMR (300 MHz, DMSO) δ 11.10 (s, 1H), 8.98 (s, 1H), 7.56 (m, 4H), 7.39 - 7.21 (m, 4H), 7.13 (t, J = 7.4 Hz, 1H), 6.83 (s, 1H), 3.22 (q, J = 12.9 Hz, 2H), 1.86 (s, 1H), 1.10 - 0.83 (m, 4H).

[0136] Example 27

[0137]

[0138] Prepared using the end product of Example 8 as the raw material: According to the method of Example 23. 1 1H NMR (300 MHz, DMSO) δ 11.14 (s, 1H), 10.52 (d, 1H), 8.98 (s, 1H), 7.77 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 8.0 Hz, 2H), 7.46 (t, J = 7.9 Hz, 2H), 7.23 (t, J = 7.2 Hz, 3H), 3.88 (d, 2H), 1.17 (s, 9H).

[0139] Example 28

[0140]

[0141] Prepared using the end product of Example 9 as the raw material: According to the method of Example 23. 1 1H NMR (300 MHz, DMSO) δ 11.14 (s, 2H), 8.99 (s, 1H), 7.88 (t, J = 2.1 Hz, 1H), 7.74 (dd, J = 8.3, 2.1 Hz, 1H), 7.66 (d, J = 8.2 Hz, 2H), 7.46 (t, J = 8.1 Hz, 1H), 7.32 - 7.17 (m, 3H), 3.65 (s, 2H), 2.08 (s, 3H).

[0142] Example 29

[0143]

[0144] Prepared using the end product of Example 10 as the raw material: According to the method of Example 23. 11H NMR(300MHz, DMSO) δ 11.14 (s, 2H), 8.98 (d, J = 1.9 Hz, 1H), 7.82 - 7.75 (m, 2H), 7.66 (d, J = 8.0 Hz, 2H), 7.54 - 7.46 (m, 2H), 7.29 (d, J = 7.8 Hz, 2H), 3.66 (s, 2H), 2.10 (s, 3H).

[0145] Example 30

[0146]

[0147] Prepared using the end product of Example 11 as the raw material: According to the method of Example 23. 1 1H NMR(300MHz, DMSO) δ 11.14 (s, 1H), 10.89 (s, 1H), 8.99 (s, 1H), 7.66 (d, J = 8.0 Hz, 2H), 7.45 - 7.25 (m, 5H), 6.77 (q, J = 4.7, 3.9 Hz, 1H), 3.78 (s, 3H), 3.64 (s, 2H), 2.08 (s, 3H).

[0148] Example 31

[0149]

[0150] Prepared using the end product of Example 12 as the raw material: According to the method of Example 23. 1 1H NMR(300MHz, DMSO) δ 11.14 (s, 2H), 8.99 (s, 1H), 8.07 (d, J = 2.4 Hz, 1H), 7.79 (dd, J = 8.9, 2.5 Hz, 1H), 7.75 - 7.57 (m, 3H), 7.29 (d, J = 8.0 Hz, 2H), 3.66 (s, 2H), 2.08 (s, 3H).

[0151] Example 32: Evaluation of the in vitro COX - 2 activity inhibition and DPPH radical scavenging activities of the example compounds

[0152] 1. Experimental method:

[0153] (1) Test method for in vitro COX - 2 enzyme activity inhibition

[0154] COX-2 used for the enzyme activity test was purchased from BPS Bioscience, and the IMAP FP Explorer Kit was purchased from Molecular Devices. The reaction mixture in the COX-2 reaction buffer (100 mM Tris-HCl, pH 8.0) contained 1.25 μg of COX-2 and indicated amounts of the control, reference inhibitor, or test inhibitor. 10 μl of the DMSO solution of the test compound was added to 100 μl of the reaction buffer, and the final concentration of DMSO was 0.5%. After incubation for 10 min, the substrate arachidonic acid (final concentration 200 μM, final ethanol concentration 1%) and the chromogenic reagent ADHP (final concentration 100 μM) were added to the reaction solution, and the reaction mixture was incubated at room temperature for 5 minutes. The fluorescence signal was measured at an excitation wavelength of 535 nM and an emission wavelength of 590 nM.

[0155] (2) In vitro DPPH free radical scavenging experiment method

[0156] DPPH (2,2-diphenyl-1-picrylhydrazyl) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. 100 μl of the methanol solution of DPPH (200 μM) was added to a 96-well plate, and then 100 μl of the methanol solution containing the compound (120 μM) or the methanol blank solvent was added. After incubation at 37 °C for 30 min, the OD value at 515 nM was measured with an enzyme-linked immunosorbent assay (ELISA) reader.

[0157] 2. Experimental results

[0158] Table 1. Results of the inhibitory activity of the compounds in the examples against COX-2 and the free radical scavenging activity of DPPH

[0159]

[0160]

[0161]

[0162]

[0163] As shown in Table 1, the compounds of the present invention all have COX-2 inhibitory activity and at the same time have free radical scavenging activity.

[0164] Example 33: Evaluation of the biological antioxidant effect of the compounds in the examples

[0165] 1. Antioxidant effect experimental method

[0166] To further clarify the antioxidant effect of the patented compound, we used a lipid peroxidation model of mouse liver tissue induced by ferrous iron as a biological oxidative stress model, with the level of lipid peroxidation product malondialdehyde (MDA) as an oxidative stress marker, calculated the inhibition rate of the compound on the production of MDA as an antioxidant activity index, and tested the antioxidant biological activity of the compound.

[0167] After the mice were sacrificed, liver tissues were taken, and liver tissue homogenates were prepared with 10% saline. Each 200 μl of tissue homogenate was used as a test sample, and the inducer FeCl 2 (1 mM), the positive control drug edaravone or the compound of Example 23 (1000 μM) were added successively, and incubated at 37 °C for 60 minutes. Then, a commercial MDA kit A003-2 (Nanjing Jiancheng Bioengineering Institute) was used to detect the level of malondialdehyde in the mouse liver homogenate according to the instructions, and the inhibition rate of the compound on the production of MDA was calculated.

[0168] 2. Experimental results

[0169] Table 2 Inhibitory effect of the compound of Example 23 on the generation of lipid peroxidation product MDA in iron-induced liver tissue

[0170]

[0171] Note: Values are expressed as mean ± SD (n = 3).

[0172] As shown in Table 2, the compound of Example 23 has significant antioxidant activity, which is superior to the free radical scavenging positive drug edaravone.

[0173] Example 34: Evaluation of the analgesic effect of the example compound

[0174] 1. Experimental method for evaluating analgesic effect

[0175] To further clarify the analgesic effect of the patented compound, we used a mouse pain model induced by intraperitoneal injection of acetic acid, with the number of writhing times of the mice as the pain observation index, calculated the inhibition rate of the compound on the number of writhing times as the analgesic effect, and tested the analgesic activity of the compound.

[0176] Mice were intraperitoneally injected with normal saline 30 minutes before acetic acid induction as a model control group, celecoxib (20 mg / kg) as a positive control group, and the compound of Example 23 (20 mg / kg) as a compound test group. Five minutes after each group of mice was intraperitoneally injected with 0.6% acetic acid solution (0.1 ml / 10 g), the number of writhing and stretching times of the mice was recorded for 15 minutes, and the inhibition rate of the compound on mouse writhing was calculated.

[0177] 2. Experimental results

[0178] Table 3. Inhibitory effect of the compound of Example 23 on acetic acid-induced pain in mice

[0179]

[0180] Note: Values are expressed as mean ± SD (n = 8).

[0181] As shown in Table 3, the compound of Example 23 has a significant analgesic effect, and its analgesic activity is similar to that of celecoxib.

Claims

1. A 1-aryl-5-pyrazolone compound, characterized in that, it has a structure selected from any of the following, and also includes its tautomer, pharmaceutically acceptable salt or a mixture thereof: wherein: R 1 selected from C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, 3-6 membered cycloalkyl, 3-6 membered halocycloalkyl; R 2 selected from hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy; R 3 selected from hydrogen, halogen; R 4 、R 5 is selected from hydrogen, -(CH 2 ) n -SO 2 -(C1-C4 alkyl), -(CH 2 ) n -CO-(C1-C4 alkyl), -(CH 2 ) n -COOH, -(CH 2 ) n -CO-NH-OH; n is an integer selected from 0 to 3.

2. The 1-aryl-5-pyrazolone compound according to claim 1, characterized in that, in the said structure: R 1 selected from C1-C4 alkyl, C1-C4 haloalkyl, 3-5 membered cycloalkyl, 3-5 membered halocycloalkyl; R 2 selected from hydrogen, halogen, C1-C4 alkoxy, C1-C4 haloalkoxy; R 3 selected from hydrogen, halogen; R 4 、R 5 selected from hydrogen, -(CH 2 ) n -SO 2 -(C1-C4 alkyl), -(CH 2 ) n -CO-(C1-C4 alkyl), -(CH 2 ) n -COOH, -(CH 2 ) n -CO-NH-OH; n is selected from 0 and 1.

3. The 1-aryl-5-pyrazolone compound according to claim 2, characterized in that, in the said structure: R 1 selected from C1-C4 alkyl, 3- to 5-membered cycloalkyl; R 2 selected from hydrogen, halogen, C1-C4 alkoxy groups; R 3 selected from hydrogen, halogen; R 4 and R 5 are selected from hydrogen, -SO 2 -(C1-C4 alkyl), -(CH 2 ) n -CO-(C1-C4 alkyl), -(CH 2 ) n -COOH, -CO-NH-OH; n is selected from 0 and 1.

4. The 1-aryl-5-pyrazolone compound according to claim 3, characterized in that, in the said structure: R 1 selected from methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl; R 2 selected from hydrogen, fluorine, chlorine, methoxy, ethoxy; R 3 selected from hydrogen, fluorine, chlorine; R 4 and R 5 are selected from hydrogen, -SO 2 -CH 3 -, -SO 2 -CH 2 CH 3 -, -(CH 2 ) n -CO-CH 3 -, -(CH 2 ) n -CO-CH 2 CH 3 -, -(CH 2 ) n -COOH, -CO-NH-OH; n is selected from 0 and 1.

5. The 1-aryl-5-pyrazolone compound according to claim 4, characterized in that, in the said structure: R 2 、R 3 When they are both disubstituted, they are both hydrogen or chlorine. R 4 and R 5 are not both hydrogen, and R 4 and R 5 are different.

6. The 1-aryl-5-pyrazolone compound according to claim 1, characterized in that, a compound selected from any of the following:

7. The 1-aryl-5-pyrazolone compound according to claim 1, characterized in that, the pharmaceutically acceptable salt is a salt formed by the said compound and an acid or a base, the acid is 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, ferulic acid, and the base is an inorganic base containing an alkaline metal cation, an alkaline earth metal cation or an ammonium cation salt.

8. A pharmaceutical composition, characterized in that, it contains the 1-aryl-5-pyrazolone compound according to claim 1 and a pharmaceutically acceptable carrier.

9. Use of the 1-aryl-5-pyrazolone compound according to claim 1 in the preparation of a COX-2 inhibitor and a free radical scavenger drug.

10. The use according to claim 9, characterized in that, the said drug is an anti-inflammatory, antioxidant stress and analgesic drug.

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