A 3-phenyl-1H-pyrazole compound and its use

Abstract

The present invention discloses a 3-phenyl-1H-pyrazole derivative as shown in general formula I or its enantiomer, diastereomer, racemate, pharmaceutically acceptable salt, and a pharmaceutical composition containing the same. Such compounds can be used as cGAS-STING pathway targeted inhibitors to target and inhibit cGAMP from stimulating the cGAS-STING pathway, and are useful for preparing drugs for treating and / or preventing inflammatory diseases and autoimmune diseases.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a 3-phenyl-1H-pyrazole compound and an application thereof. The compound has strong STING inhibitory activity in stably transfected THP1 cells. Background Art

[0002] Stimulator of interferon gene (STING) is a protein encoded by the human TMEM173 gene and plays an important regulatory role in the innate immune system. The innate immune system recognizes pathogens through pattern recognition receptors (PRRs) and upregulates the immune function of immune cells, while inducing the secretion of a large number of inflammatory factors and chemokines, thereby activating adaptive immunity. Among them, pattern recognition receptors include the upstream protein of STING, guanylate-adenylate synthetase (Cyclic GMP-AMP synthase, cGAS). As an intracellular DNA sensor, cGAS can recognize exogenous substances or its own cytoplasmic double-stranded DNA (dsDNA), synthesize and release cyclic dinucleotides (CDNs) such as c-di-GMP, c-di-AMP, 3',3'-cGAMP and 2',3'-cGAMP, etc., which activate STING and transfer it from the endoplasmic reticulum to the Golgi apparatus, and then recruit TANK binding kinase-1 (TBK-1) in the cytoplasm, mediate TBK-1 phosphorylation of interferon regulatory factor-3 (IRF-3), induce the secretion of type I interferon, and induce the secretion of multiple inflammatory factors such as TNF, IL-6 and CXCL10. In addition, STING also induces the production of pro-inflammatory factors through the NF-κB pathway.

[0003] Excessive activation of STING can lead to a series of inflammatory diseases. These diseases include STING-associated vasculopathy withonset in infancy (SAVI) syndrome, Aicardi-Goutières syndrome (AGS), and hereditary lupus caused by gain-of-function mutations in TMEM173. In addition to these genetic diseases, increasing evidence suggests that STING has a broader pathogenic role in a series of inflammatory-related diseases (such as systemic lupus erythematosus, rheumatoid arthritis, nerve damage, renal fibrosis, systemic sclerosis, and some cancer metastases). Therefore, small molecule-based drug intervention in the STING signaling pathway has important potential for the treatment of diseases caused by excessive STING activation.

[0004] WO2020010155A1 discloses a STING inhibitor, which has the following structural formula:

[0005]

[0006] The inhibitory effect of this type of compound on STING is at the micromolar level. Therefore, more STING inhibitors with new structures need to be developed. Summary of the invention

[0007] Objective of the invention: One of the objectives of the present invention is to provide a 3-phenyl-1H-pyrazole compound of general formula I or its enantiomers, diastereomers, racemates and pharmaceutically acceptable salts thereof:

[0008]

[0009] in,

[0010] X1, X2 are each independently selected from halogen or hydrogen;

[0011] R1 is selected from hydrogen or methyl;

[0012] A is selected from a 6-10 membered aromatic ring, aromatic heterocyclic ring or condensed ring substituted by R2;

[0013] R2 is selected from hydrogen, halogen, halogen-substituted or unsubstituted C1-C9 alkyl or heteroalkyl, halogen-substituted or unsubstituted C1-C6 alicyclic or alicyclic heterocyclic;

[0014] L is selected from -U1-NH-(U2) n1 -(U3) n2 -;

[0015] U1, U2, U3 are each independently selected from -CH2- or -(C=O)-;

[0016] n1 and n2 are each independently selected from 0 or 1.

[0017] In certain preferred embodiments,

[0018] A is selected from R2 substituted benzene ring, indole or benzocyclohexane.

[0019] In certain preferred embodiments,

[0020] R2 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, trifluoromethyl, N,N-dimethylamino, trifluoromethylthio, n-hexyloxy, N-methylpiperazinyl, N-methylpiperazinylmethyl or cyclohexyl.

[0021] The compounds of the general formula I of the present invention are preferably the following compounds:

[0022]

[0023]

[0024] The above-mentioned compounds of general formula I of the present invention may also exist in the form of their salts, which are converted in vivo into compounds of general formula I. For example, within the scope of the present invention, the compounds of the present invention are converted into pharmaceutically acceptable salt forms according to processes known in the art, and they are used in the form of salts.

[0025] All tautomeric forms of the compounds of the general formula I of the present invention are included within the scope of the present invention. The compounds of the present invention may exist in specific geometric or stereoisomer forms. Additional asymmetric carbon atoms may exist in substituents such as alkyl, and all these isomers and their mixtures are included within the scope of the present invention.

[0026] In some preferred embodiments, the pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed by the compound of formula I and the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, maleic acid, benzenesulfonic acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, fumaric acid, salicylic acid or phenylacetic acid; and also include acid salts formed by the compound of formula I and inorganic bases or organic salts made from basic amines.

[0027] The present invention also provides a synthetic route for compounds having the general formula I:

[0028] (1) When R1 is selected from hydrogen, L is selected from -U1-NH-(U2) n1 -(U3) n2-; U1, U2, U3 are each independently selected from -CH2- or -(C=O)-, n1, n2 are each independently selected from 0 or 1, the compound synthesis route is as follows:

[0029]

[0030] Reagents and conditions: (a) acetonitrile, sodium hydride, toluene, 90°C, 4h; (b) N,N-dimethylformamide dimethyl acetal, 90°C, 3h; (c) ethanol, hydrazine hydrate, 70°C, 3h; (d) lithium aluminum hydride, tetrahydrofuran, 25°C, 10h; (e) A-COOH, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N-diisopropylethylamine, N,N-dimethylformamide, 25°C, 8h; (f) A-CHO, sodium borohydride, anhydrous dichloromethane, anhydrous magnesium sulfate, 25°C, 10h.

[0031] (2) When R1 is selected from hydrogen, L is selected from -U1-NH-(U2) n1 -(U3) n2 -; U1, U2, U3 are each independently selected from -CH2- or -(C=O)-, n1, n2 are selected from 0, X1, X2 are each independently selected from hydrogen, the compound synthesis route is as follows:

[0032]

[0033] Reagents and conditions: (h) 1) catalytic amount of glacial acetic acid, ethanol, reflux, 4h; 2) trifluoroacetic acid, dichloromethane, 25℃, 0.5h; 3) phosphorus oxychloride, N,N-dimethylformamide, ice bath for 1h, room temperature for 0.5h, heating at 80℃ for 4h; (i) potassium permanganate, potassium hydroxide, 30% hydrogen peroxide, dioxane / water, 25℃, 3h; (j) A-NH2, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N-diisopropylethylamine, N,N-dimethylformamide, 25℃, 8h; (k) sodium borohydride, anhydrous dichloromethane, anhydrous magnesium sulfate, 25℃, 10h.

[0034] In the above synthetic route, X1, X2 and A are defined as shown in general formula I.

[0035] The compounds of the general formula I of the present invention can be prepared by the above or similar preparation methods, and the corresponding starting materials can be selected according to the different substituents and the different positions of the substituents. It should be appreciated by those skilled in the art that the above routes are helpful for understanding the present invention, but do not limit the content of the present invention. Unless otherwise specified, the variables are defined as mentioned in the general formula I.

[0036] Another object of the present invention is to provide a pharmaceutical composition, which comprises a compound of formula I or a pharmaceutically acceptable salt, crystal form, hydrate, solvate, prodrug, racemate, metabolite thereof and a pharmaceutically acceptable carrier.

[0037] The pharmaceutical composition of the present invention can be administered in various known ways, such as orally, parenterally, by inhalation spray or via an implanted reservoir. The pharmaceutical composition of the present invention can be administered alone or in combination with other drugs. Oral compositions can be in any orally acceptable dosage form, including but not limited to tablets, capsules, emulsions, suspensions, dispersions and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, adhesives, colorants, fillers, emulsifiers, etc.

[0038] Sterile injectable compositions can be formulated using suitable dispersing agents or wetting agents and suspending agents according to techniques known in the art. Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride solution, and the like.

[0039] Topical compositions may be formulated as oils, lotions, creams, etc. Carriers for the compositions include vegetable or mineral oils, animal fats, and high molecular weight alcohols, etc. Pharmaceutically acceptable carriers are carriers in which the active ingredient is soluble.

[0040] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The dosage level selected depends on a variety of factors, including the activity of the specific compound of the present invention or its salt used, the route of administration, the time of administration, the excretion rate of the specific composition used, the duration of treatment, other drugs, compounds and / or materials used in combination with the specific composition used, the age, sex, weight, general health and previous medical history of the patient being treated, and similar factors well known in the medical field.

[0041] Another object of the present invention is to provide the use of the compound of formula I or its pharmaceutically acceptable salt, enantiomer, diastereomer, racemate thereof in the preparation of drugs for treating diseases related to the function of STING protein.

[0042] The disease related to the function of STING protein is a disease caused by excessive activation of the STING pathway.

[0043] Another object of the present invention is to provide use of the compound of general formula I in the preparation of a medicament for preventing and / or treating inflammatory diseases and autoimmune diseases.

[0044] The diseases, inflammatory diseases and autoimmune diseases related to the function of STING protein include STING-related infantile vasculopathy SAVI, Aicardi-Goutières syndrome AGS, COPA syndrome caused by gene mutation of the α subunit of the symbiotic protein complex, systemic lupus erythematosus SLE, familial pernio lupus erythematosus FCL, Parkinson's disease PD, amyotrophic lateral sclerosis ALS, Huntington's disease, non-alcoholic steatohepatitis NASH, alcoholic liver, nerve damage, rheumatoid arthritis, renal fibrosis, systemic sclerosis, intervertebral disc degeneration, pulmonary fibrosis, aging, scleroderma, psoriasis, inflammatory bowel disease, autoimmune colitis, irritable bowel syndrome, ulcerative colitis, Crohn's disease, uveitis, mucositis, diabetes, cardiovascular disease or neurodegenerative disease.

[0045] Beneficial effects:

[0046] The present invention synthesized a 3-phenyl-1H-pyrazole compound based on the skeleton transition of the lead compound. The compound exhibited good STING inhibitory activity and had strong STING inhibitory activity in stably transfected THP1 cells. The compound can be used to prepare drugs for treating diseases related to the function of the STING protein, and has the prospect of preparing drugs for treating and / or preventing inflammatory diseases and autoimmune diseases, providing a reliable approach for clinical application. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 This is the ITC titration curve of compound 5 and STING protein.

[0048] Figure 2 This is the ITC titration curve of compound 22 and STING protein. DETAILED DESCRIPTION

[0049] The preparation method of the compound of general formula I of the present invention is described below in conjunction with specific examples, but these specific methods do not constitute any limitation to the present invention. The compound of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such a combination can be easily carried out by a technician in the field to which the present invention belongs.

[0050] The starting materials, reaction reagents, etc. used in the specific embodiments of the present invention are all commercially available. The present invention can be prepared in the form of salt by using a salt-forming method commonly used in the art, for example, dissolving the compound in hydrochloric acid ethanol at room temperature to react to generate a hydrochloride; or adding benzenesulfonic acid to react to generate a benzenesulfonate.

[0051] The experimental methods in the examples of the present invention where specific conditions are not specified are generally carried out under conventional conditions or under conditions recommended by raw material or product manufacturers.

[0052] The melting points of all compounds in the examples were determined by Thomas Hoover apparatus. The structures of the compounds were determined by H NMR ( 1 HNMR) was measured by Bruker Avance 300 MHz spectrometer using TMS as internal standard and recording temperature at 300 K. The purity of the compound was evaluated by reverse phase HPLC (Agilent 1260, Hanbang C18 4.6×150 mm, 5 μm) with an elution system of methanol: water = 80:20 (v / v).

[0053] Unless otherwise specified in the examples, all reactions were carried out under air atmosphere.

[0054] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0055] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20-30°C.

[0056] Example 1

[0057] 4-n-Butyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)aniline (1)

[0058]

[0059] Step 1: Preparation of compound 1-3

[0060] Compound 1-1 (500 mg, 4.17 mmol) was dissolved in ethanol (20 mL), and compound 1-2 (604 mg, 4.58 mmol) and glacial acetic acid (24 μL, 0.417 mmol) were added respectively. The reaction solution was evaporated after heating at 80 ° C for 4 h. The remaining solid was dissolved in dichloromethane (20 mL), trifluoroacetic acid (5 mL) was added, and the solvent was evaporated to obtain a white solid for use. Phosphorus oxychloride (1 mL) was added dropwise to N, N-dimethylformamide (3 mL) under ice bath, and the ice bath was stirred for 1 h. The obtained white solid was added to the reaction solution under ice bath, stirred at room temperature for 0.5 h, and heated to 80 ° C for 4 h. After the reaction was completed, the reaction solution was poured into ice water and neutralized with saturated sodium bicarbonate aqueous solution until no carbon dioxide gas was generated. The mixture was extracted three times with ethyl acetate / water, and the organic phases were combined and washed with saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, spin-dried, and chromatographed on a silica gel column (petroleum ether:ethyl acetate=50:1-1:1, v / v) to obtain 216 mg of a light yellow solid with a yield of 30%. 1H NMR (300MHz, Chloroform-d) δ9.96(s,1H,CHO),9.67(s,1H,N-NH),8.10(s,1H,NH-CH),7.75-7.59(m,2H,Ar-H),7.58-7.44(m,3H,Ar-H).

[0061] Step 2: Preparation of compound 1

[0062] Dissolve p-butylaniline (100 mg, 0.671 mmol) in anhydrous dichloromethane (5 mL), add compound 1-3 (138 mg, 0.805 mmol) and anhydrous magnesium sulfate (800 mg, 500 mg / mmol) respectively. Stir at room temperature for 8 h under nitrogen protection. Evaporate the reaction solution, redissolve the remaining solid in methanol (5 mL), add sodium borohydride (51 mg, 1.342 mmol) under ice bath, and stir at room temperature for 2 h. After the reaction is complete, add water to quench the reaction, evaporate methanol, extract (ethyl acetate / water) three times, combine the organic phases, wash the organic phase with saturated sodium chloride aqueous solution, dry the organic phase with anhydrous sodium sulfate, spin dry, and silica gel column chromatography (dichloromethane: methanol = 50: 1, v / v) to obtain 187 mg of white solid, with a yield of 91.4%. Melting point: 96.9-98.1 ° C. Purity: 99.65%.

[0063] 1 H NMR(300MHz,DMSO-d6)δ13.24-12.64(m,1H,N-NH),7.89-7.17(m,6H,Ar-H),6.94-6.75(m,2H,Ar-H),6.62-6.39(m,2H,Ar-H) ,5.56(s,1H,Ar-NH),4.08(s,2H,NHCH2),2.36(m,2H,Ar-CH2),1.42(m,2H,CH2),1.23(m,2H,CH2),0.83(t,J=7.1Hz,3H,CH3).

[0064] Example 2

[0065] N-(4-Butylphenyl)-3-phenyl-1H-pyrazole-4-carboxamide (2)

[0066]

[0067] Step 1: Preparation of compound 1-4

[0068] Compound 1-3 (100 mg, 0.58 mmol) was dissolved in dioxane (4 mL), water (1 mL) was added, potassium hydroxide (39 mg, 0.7 mmol) and potassium permanganate (110 mg, 0.7 mmol) were added under ice bath, and the mixture was transferred to room temperature and stirred for 3 h. After the reaction was completed, 30% hydrogen peroxide (2 mL) was slowly added dropwise under ice bath. The precipitate was filtered, the filtrate was adjusted to pH 2 with dilute hydrochloric acid, extracted (ethyl acetate / water) three times, the organic phases were combined, the organic phases were washed with saturated sodium chloride aqueous solution, the organic phases were dried over anhydrous sodium sulfate, and spin dried. The solid was slurried with an organic solvent (petroleum ether: ethyl acetate = 20: 1, v / v), filtered, and the filter cake was dried under an infrared lamp to obtain 104 mg of a white solid with a yield of 95%. 1 H NMR (300MHz, DMSO-d6) δ12.82(brs,2H,COOH,N-NH),8.14(s,1H,NH-CH),7.80-7.68(m,2H,Ar-H),7.44(m,3H,Ar-H).

[0069] Step 2: Preparation of compound 2

[0070] Compound 1-4 (100 mg, 0.531 mmol) was dissolved in N, N-dimethylformamide (3 mL), and O-(7-azabenzotriazole-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate (303 mg, 0.796 mmol) and N, N-diisopropylethylamine (139 μL, 103 mmol) were added respectively. After stirring at room temperature for 0.5 h, a solution of p-n-butylaniline (102 μL, 0.637 mmol) in N, N-dimethylformamide (3 mL) was added and stirred at room temperature for 8 h. After the reaction was completed, water was added to the reaction solution, and the mixture was extracted (ethyl acetate / water) three times. The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and dried by spin drying. The mixture was subjected to silica gel column chromatography (dichloromethane: methanol = 150: 1–50: 1, v / v) to obtain 85 mg of a light yellow solid with a yield of 50.29%. Purity: 98.24%.

[0071] Melting point: 153.8-155.6℃.

[0072] 1H NMR(300MHz,DMSO-d6)δ13.57-13.11(m,1H,N-NH),9.84(s,1H,NHCH),8.33-7.95(m,1H,CONH),7.76-7.59(m,2H,Ar-H),7.51(d,J=7.6Hz,2H,Ar-H),7.4 6-7.21(m,3H,Ar-H),7.07(d,J=7.9Hz,2H,Ar-H),2.56-2.47(m,2H,Ar-CH2) ,1.57-1.39(m,2H,CH2),1.33-1.16(m,2H,CH2),0.84(t,J=7.0Hz,3H,CH2).

[0073] Example 3

[0074] N-(4-Butylbenzyl)-3-phenyl-1H-pyrazole-4-carboxamide (3)

[0075]

[0076] Step 1: Preparation of compound 2-2

[0077] Compound 2-1 (200 mg, 1.26 mmol) was dissolved in tetrahydrofuran (5 mL), and lithium aluminum hydride (238 mg, 6.28 mmol) was added in batches under an ice bath and stirred at room temperature for 2 h. After the reaction was complete, water (238 μL) was added under an ice bath to quench the reaction, and the ice bath was stirred for 10 min. A 10% aqueous sodium hydroxide solution (2 mL) was added under an ice bath, and the ice bath was stirred for 10 min. Water (3 mL) was added under an ice bath, and the precipitate was filtered after stirring for 10 min (with diatomaceous earth as the filter aid). After the tetrahydrofuran in the filtrate was dried, ethyl acetate was added to the filtrate and extracted three times, the organic phases were combined, the organic phases were washed with a saturated sodium chloride aqueous solution, the organic phases were dried over anhydrous sodium sulfate, dried, and chromatographed on a silica gel column (dichloromethane: methanol = 20: 1, v / v) to obtain 170 mg of an oily product with a yield of 82.9%. 1 H NMR(300MHz,Chloroform-d)δ7.19(d,J=7.5Hz,2H,Ar-H),7.15(d,J=7.5Hz,2H,Ar-H),3.92(t,J=6.7Hz,2H,NH2CH2),2 .58(t,J=7.3Hz,2H,CH2),2.16(s,2H,NH2),1.60-1.51(m,2H,CH2),1.39-1.27(m,2H,CH2),0.96(t,J=8.0Hz,3H,CH3).

[0078] Step 2: Preparation of compound 3

[0079] The preparation method is the same as step 2 in Example 2, except that p-n-butylaniline is replaced with compound 2-2 (104 mg, 0.638 mmol), to obtain 80 mg of a light yellow solid with a yield of 45.2%. Melting point: 134.1-136.4°C. Purity: 99.27%.

[0080] 1 H NMR(300MHz,DMSO-d6)δ13.49-12.98(m,1H,N-NH),8.47(s,1H,NHCH),8.21-7.86(m,1H,CONH),7.65(d,J=2.0Hz,2H,Ar-H),7.44-7.19(m,3H,Ar-H),7.2 1-7.00(m,4H,Ar-H),4.29(d,J=5.0Hz,2H,NHCH2),2.53-2.46(m,2H,Ar-CH2 ),1.56-1.35(m,2H,CH2),1.34-1.09(m,2H,CH2),0.82(t,J=6.3Hz,3H,CH3).

[0081] Example 4

[0082] (3-phenyl-1H-pyrazol-4-yl)methanamine (3-4)

[0083]

[0084] Step 1: Preparation of compound 3-2

[0085] Compound 3-1 (2 g, 13.79 mmol) was dissolved in N,N-dimethylformamide dimethyl acetal (5 mL) and heated to reflux at 90 °C for 3 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate and the organic phase was washed with a saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous sodium sulfate, spin-dried, and chromatographed on a silica gel column (petroleum ether: ethyl acetate = 1:1, v / v) to obtain 1.5 g of a white solid with a yield of 54.54%. 1 HNMR (300MHz, Chloroform-d) δ7.95 (s, 1H, Ar-H), 7.78 (d, J = 7.0Hz, 2H, Ar-H), 7.55-7.37 (m, 3H, Ar-H), 3.49 (s, 3H, CH3), 3.29 (s, 3H, CH3).

[0086] Step 2: Preparation of compound 3-3

[0087] Compound 3-2 (1.5 g, 7.5 mmol) was dissolved in ethanol (10 mL) and 80% hydrazine hydrate (455 μL, 7.5 mmol) was added, and the mixture was heated to reflux at 70°C for 3 h. After the reaction was completed, the solvent was evaporated and the solid was redissolved in ethyl acetate. The organic phase was washed 3 times with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and dried by spin drying. The mixture was chromatographed on a silica gel column (petroleum ether: ethyl acetate: dichloromethane = 2:1:1, v / v) to obtain 1 g of a light yellow solid with a yield of 78.93%. 1 HNMR(300MHz,Chloroform-d)δ8.73(brs,1H,NH),7.99(s,1H,Ar-H),7.90-7.81(m,2H,Ar-H),7.56-7.46(m,3H,Ar-H).

[0088] Step 3: Preparation of compound 3-4

[0089] The preparation method is the same as step 1 in Example 3, except that compound 2-1 is replaced by compound 3-3 (500 mg, 2.96 mmol), and the amount of lithium aluminum hydride is 561 mg (14.8 mmol). Silica gel column chromatography (dichloromethane: methanol = 100: 1-20: 1, v / v) is performed to obtain 216 mg of a light yellow solid, with a yield of 42.18%. 1 HNMR(300MHz,Chloroform-d)δ7.64-7.48(m,3H,Ar-H),7.48-7.29(m,3H,Ar-H),4.80(brs,3H,NH,NH2),3.92(s,2H,CH2).

[0090] Example 5

[0091] 4-Butylbenzaldehyde

[0092]

[0093] Step 1: Preparation of methyl p-butylbenzoate

[0094] Dissolve p-butylbenzoic acid (500 mg, 2.8 mmol) in methanol (10 mL), slowly add dichlorothionyl (2 mL, 28 mmol) dropwise under ice bath, after addition, protect with nitrogen, heat to 60 ° C and reflux for 3 h. After the reaction, evaporate the solvent, add saturated sodium bicarbonate under ice bath to neutralize until no carbon dioxide gas is generated. Extract (ethyl acetate / water) three times, combine the organic phases, wash the organic phase with saturated sodium chloride aqueous solution, dry the organic phase with anhydrous sodium sulfate, spin dry, and chromatograph on a silica gel column (petroleum ether: ethyl acetate = 20:1, v / v). 530 mg of white solid was obtained, with a yield of 98.69%. 1H NMR(300MHz,Chloroform-d)δ8.05-7.86(m,2H,Ar-H),7.35-7.17(m,2H,Ar-H),3.92(s,3H,OCH3),2.68(t, J=7.7Hz,2H,CH2),1.63(p,J=7.5Hz,2H,CH2),1.39(h,J=14.8,7.3Hz,2H,CH2),0.95(t,J=7.3Hz,3H,CH3).

[0095] Step 2: Preparation of 4-butylbenzyl alcohol

[0096] The preparation method is the same as step 1 in Example 3, except that compound 2-1 is replaced with methyl p-butylbenzoate (530 mg, 2.76 mmol), and the amount of lithium aluminum hydride used is 461 mg (12.14 mmol). Silica gel column chromatography (petroleum ether: ethyl acetate = 5:1, v / v) is performed to obtain 430 mg of an oily product with a yield of 95.13%. 1 H NMR(300MHz,Chloroform-d)δ7.23(d,J=7.5Hz,2H,Ar-H),7.11(d,J=7.5Hz,2H,Ar-H),4.64(d,J=6.4Hz,2H,CH2),4.13 -3.96(m,1H,OH),2.60(t,J=7.0Hz,2H,CH2),1.61-1.52(m,2H,CH2),1.39-1.27(m,2H,CH2),0.96(t,J=8.0Hz,3H,CH3).

[0097] Step 3: Preparation of p-butylbenzaldehyde

[0098] Dissolve p-butylbenzyl alcohol (430 mg, 2.62 mmol) in anhydrous dichloromethane (5 mL), add dropwise to an anhydrous dichloromethane solution (5 mL) of pyridinium chlorochromate (1.13 g, 5.24 mmol), protect with nitrogen, and stir at room temperature for 2 h. After the reaction is completed, filter, extract the filtrate with water, and wash the organic phase with 1M sodium hydroxide solution and 1M dilute hydrochloric acid, respectively. Spin dry the organic phase, and chromatograph on a silica gel column (petroleum ether: ethyl acetate = 20:1, v / v). 400 mg of oily product is obtained, with a yield of 94.34%. 1H NMR(300MHz,Chloroform-d)δ9.97(s,1H,-CHO),7.80(d,J=8.0Hz,2H,Ar-H),7.34(d,2H,Ar-H),2 .69(t,2H,CH2), 1.63(p,J=7.5Hz,2H,CH2), 1.36(h,J=7.3Hz,2H,CH2), 0.93(t,J=7.3Hz,3H,CH3).

[0099] Example 6

[0100] N-(4-Butylbenzyl)-1-(3-phenyl-1H-pyrazol-4-yl)methanamine (4)

[0101]

[0102] The preparation method is the same as step 2 in Example 1, except that p-butylaniline is replaced by compound 3-4 (100 mg, 0.578 mmol), and compound 1-3 is replaced by p-butylbenzaldehyde (112 mg, 0.693 mmol). Silica gel column chromatography (petroleum ether: ethyl acetate = 1:1, v / v) gives 162 mg of a white solid, with a yield of 88.04%. Melting point: 83.0-58.2°C. Purity: 96.10%.

[0103] 1 H NMR(300MHz,DMSO-d6)δ13.13-12.63(m,1H,N-NH),7.71(s,3H,Ar-H,NHCH),7.47 -7.27(m,3H,Ar-H),7.24(d,J=7.8Hz,2H,Ar-H),7.12(d,2H,Ar-H),3.70(s,2H,A r-CH2-NH),3.61(s,2H,Ar-CH2-NH),2.60-2.51(m,2H,Ar-CH2),2.38(brs,1H,CH 2-NH), 1.61-1.45 (m, 2H, CH2), 1.36-1.21 (m, 2H, CH2), 0.88 (t, J = 7.3Hz, 2H, CH3).

[0104] Example 7

[0105] 4-Butyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (5)

[0106]

[0107] The preparation method is the same as step 2 in Example 2, except that p-n-butylaniline is replaced by compound 3-4 (100 mg, 0.578 mmol), and compound 1-4 is replaced by p-n-butylbenzoic acid (85 mg, 0.481 mmol). 112 mg of light yellow solid is obtained, with a yield of 70%. Melting point: 167.5-169.8°C. Purity: 95.50%.

[0108] 1 H NMR(300MHz,Chloroform-d)δ7.64(s,1H,NHCH),7.62-7.53(m,4H,Ar-H).7.47-7.32(m,3H,Ar-H),7.19(d,J=7.9Hz,2H,Ar-H),6.91(brs,1H,N- NH), 6.28 (t, J = 5.2Hz, 1H, CONH), 4.65 (d, J = 5.1Hz, 2H, CH2NH), 2.62 (m, 2H, CH2), 1.58 (m, 2H, CH2), 1.33 (m, 2H, CH2), 0.91 (t, J = 7.3Hz, 3H, CH3).

[0109] Example 8

[0110] 4-(tert-Butyl)-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (6)

[0111]

[0112] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (80 mg, 0.462 mmol), and compound 1-4 is replaced by p-tert-butylbenzoic acid (69 mg, 0.385 mmol). 67 mg of white solid is obtained, with a yield of 52.34%. Melting point: 203.6-206.4°C. Purity: 99.62%.

[0113] 1 H NMR(300MHz,DMSO-d6)δ8.80(t,J=4.9Hz,1H,CONH),7.86-7.76(m,2H,Ar-H),7.67(dd,J=9.3,1.8Hz,3H,Ar -H),7.53-7.42(m,4H,Ar-H),7.42-7.31(m,1H,Ar-H),4.48(d,J=5.3Hz,2H,NHCH2),1.30(s,9H,(CH3)3C).

[0114] Example 9

[0115] 4-Isopropyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (7)

[0116]

[0117] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (64 mg, 0.366 mmol), and compound 1-4 is replaced by p-isopropylbenzoic acid (50 mg, 0.305 mmol). 53 mg of white solid is obtained, with a yield of 54.63%. Melting point: 195.1-196.7°C. Purity: 99.62%.

[0118] 1 H NMR (300MHz, DMSO-d6) δ13.11-2.78(m,1H,NNH),8.80(s,1H,CONH),7.81(d,J=8.2Hz,2H,Ar-H),7.74-7.62(m,2H,Ar -H),7.54-7.24(m,6H,Ar-H),4.48(s,2H,NHCH2),2.94(p,J=6.9Hz,1H,(CH3)2CH),1.22(d,J=6.9Hz,6H,(CH3)2CH).

[0119] Example 10

[0120] 4-Isobutyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (8)

[0121]

[0122] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (53 mg, 0.308 mmol), and compound 1-4 is replaced by p-isobutylbenzoic acid (50 mg, 0.280 mmol). 47 mg of white solid is obtained, with a yield of 50.53%. Melting point: 213.6-216.0°C. Purity: 99.20%.

[0123] 1 H NMR (300MHz, DMSO-d6) δ13.11-12.73(m,1H,NNH),8.78(s,1H,CONH),7.78(d,J=8.2Hz,2H,Ar-H),7.74-7.28(m,6H,Ar-H),7.23(d,J =8.1Hz,2H,Ar-H),4.47(s,2H,NHCH2),2.49-2.45(m,2H,Ar-CH2),1.85(dp,J=13.6,6.5Hz,1H,CH),0.85(d,J=6.6Hz,6H,CH(CH3)2).

[0124] Embodiment 11

[0125] N-((3-phenyl-1H-pyrazol-4-yl)methyl)-4-propylbenzamide (9)

[0126]

[0127] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (63 mg, 0.365 mmol), and compound 1-4 is replaced by p-propylbenzoic acid (50 mg, 0.304 mmol). 56 mg of white solid is obtained, with a yield of 57.73%. Melting point: 183.9-186.0°C. Purity: 99.62%.

[0128] 1 H NMR (300MHz, DMSO-d6) δ13.10-12.67(m,1H,NNH),8.72(s,1H,CONH),7.72(d,J=8.2Hz,2H,Ar-H),7.69-7.25(m,6H,Ar-H),7.20( d, J=8.2Hz, 2H, Ar-H), 4.42 (s, 2H, NHCH2), 2.53 (t, J=7.5Hz, 2H, Ar-CH2), 1.54 (h, J=7.3Hz, 2H, CH2), 0.83 (t, J=7.3Hz, 3H, CH3).

[0129] Example 12

[0130] 4-Ethyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (10)

[0131]

[0132] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (66 mg, 0.383 mmol), and compound 1-4 is replaced by p-ethylbenzoic acid (50 mg, 0.333 mmol). 41 mg of white solid is obtained, with a yield of 40.59%. Melting point: 189.6-192.9°C. Purity: 99.79%.

[0133] 1H NMR (300MHz, DMSO-d6) δ13.10-12.67(m,1H,NNH),8.72(s,1H,CONH),7.72(d,J=8.2Hz,2H,Ar-H),7.69-7.25(m,6H,Ar-H),7.20( d, J=8.2Hz, 2H, Ar-H), 4.42 (s, 2H, NHCH2), 2.53 (t, J=7.5Hz, 2H, Ar-CH2), 1.54 (h, J=7.3Hz, 2H, CH2), 0.83 (t, J=7.3Hz, 3H, CH3).

[0134] Example 13

[0135] 4-(Dimethylamino)-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (11)

[0136]

[0137] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (63 mg, 0.363 mmol), and compound 1-4 is replaced by p-dimethylaminobenzoic acid (50 mg, 0.303 mmol). 56 mg of white solid is obtained, with a yield of 57.75%. Melting point: 216.3-219.6°C. Purity: 95.57%.

[0138] 1 H NMR(300MHz,DMSO-d6)δ13.13-12.67(m,1H,NNH),8.60-8.37(m,1H,CONH),7.74(d,J=8.9Hz,2H,Ar-H ),7.71-7.29(m,6H,Ar-H),6.74-6.64(m,2H,Ar-H),4.59-4.31(m,2H,NHCH2),2.96(s,6H,(CH3)2N).

[0139] Embodiment 14

[0140] 4-Methyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (12)

[0141]

[0142] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (76 mg, 0.44 mmol), and compound 1-4 is replaced by p-toluic acid (50 mg, 0.367 mmol). 58 mg of white solid is obtained, with a yield of 54.35%. Melting point: 181.2-185.4°C. Purity: 99.57%.

[0143] 1 H NMR(300MHz,DMSO-d6)δ13.13-12.73(m,1H,NNH),8.77(s,1H,CONH),7.76(d,J=8.1Hz,2H,Ar- H),7.73-7.29(m,6H,Ar-H),7.25(d,J=8.0Hz,2H,Ar-H),4.46(s,2H,NHCH2),2.34(s,3H,CH3).

[0144] Embodiment 15

[0145] 4-(4-Methylpiperazin-1-yl)-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (13)

[0146]

[0147] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (47 mg, 0.271 mmol), and compound 1-4 is replaced by p-methylpiperazine benzoic acid (50 mg, 0.226 mmol). 46 mg of light yellow solid is obtained, with a yield of 54.25%. Melting point: 183.2-186.1°C. Purity: 96.91%.

[0148] 1 H NMR(300MHz,DMSO-d6)δ13.11-12.72(m,1H,NNH),8.56(s,1H,CONH),7.75(d,J=8.8Hz,2H,Ar-H),7.72-7.29(m,6H,Ar-H),6.9 4(d,J=8.9Hz,2H,Ar-H),4.45(s,2H,NHCH2),3.28-3.18(m,4H,Ar-N(CH2)2),2.49-2.40(m,4H,CH3N(CH2)2),2.23(s,3H,CH3).

[0149] Example 16

[0150] 4-((4-Methylpiperazin-1-yl)methyl)-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (14)

[0151]

[0152] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (45 mg, 0.256 mmol), and compound 1-4 is replaced by p-methylpiperazinylmethylbenzoic acid (50 mg, 0.213 mmol). 41 mg of light yellow solid is obtained, with a yield of 49.51%. Melting point: 164.5-167.9°C. Purity: 96.10%.

[0153] 1 H NMR (300MHz, DMSO-d6) δ13.17-12.73(m,1H,NNH),8.82(s,1H,CONH),7.83(d,J=7.9Hz,2H,Ar-H),7.78- 7.27(m,8H,Ar-H),4.49(s,2H,NHCH2),3.52(s,2H,Ph-CH2),2.42(s,8H,Pip(CH2)4),2.25(s,3H,CH3).

[0154] Embodiment 17

[0155] 4-Pentyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (15)

[0156]

[0157] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (54 mg, 0.312 mmol), and compound 1-4 is replaced by p-pentylbenzoic acid (50 mg, 0.26 mmol). 61 mg of light yellow solid is obtained, with a yield of 67.61%. Melting point: 140.0-143.5°C. Purity: 99.47%.

[0158] 1 H NMR (300MHz, DMSO-d6) δ13.19-12.74(m,1H,NNH),8.76(s,1H,CONH),7.78(d,J=8.1Hz,2H,Ar-H),7.74-7.30(m,6H,Ar-H),7.26(d,J=8.1Hz,2 H, Ar-H), 4.47 (s, 2H, NHCH2), 2.61 (t, J = 7.6Hz, 2H, Ar-CH2), 1.57 (p, J = 7.3Hz, 2H, CH2), 1.40-1.15 (m, 4H, (CH2)2), 0.85 (t, J = 6.8Hz, 3H, CH3).

[0159] Embodiment 18

[0160] 4-Heptyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (16)

[0161]

[0162] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (47 mg, 0.272 mmol), and compound 1-4 is replaced by p-heptylbenzoic acid (50 mg, 0.227 mmol). 41 mg of light yellow solid is obtained, with a yield of 48.14%. Melting point: 132.7-137.3°C. Purity: 99.00%.

[0163] 1 H NMR(300MHz,DMSO-d6)δ13.14-12.73(m,1H,NNH),8.79(t,J=4.6Hz,1H,CONH),7.78 (d,J=8.1Hz,2H,Ar-H),7.67(d,J=7.4Hz,3H,Ar-H),7.52-7.31(m,3H,Ar-H),7.26(d ,J=8.1Hz,2H,Ar-H),4.47(d,J=4.9Hz,2H,NHCH2),2.60(t,J=7.5Hz,2H,Ar-CH2),1. 56(p,J=7.6,6.8Hz,2H,CH2),1.33-1.17(m,8H,(CH2)4),0.85(t,J=6.5Hz,3H,CH3).

[0164] Embodiment 19

[0165] 4-Cyclohexyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (17)

[0166]

[0167] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (51 mg, 0.294 mmol), and compound 1-4 is replaced by p-cyclohexylbenzoic acid (50 mg, 0.245 mmol). 51 mg of light yellow solid is obtained, with a yield of 57.95%. Melting point: 224.6-231.3°C. Purity: 98.99%.

[0168] 1H NMR(300MHz,DMSO-d6)δ13.18-12.73(m,1H,NNH),8.75(s,1H,CONH),7.78(d,J=8.2Hz,2H,Ar-H),7.74-7.32(m,6H,Ar-H) ,7.29(d,J=8.2Hz,2H,Ar-H),4.47(s,2H,CH2),2.62-2.53(m,1H,Ar-CH),1.85-1.64(m,5H,CH2),1.50-1.18(m,5H,CH2).

[0169] Embodiment 20

[0170] N-((3-phenyl-1H-pyrazol-4-yl)methyl)-4-(trifluoromethylthio)benzamide (18)

[0171]

[0172] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (50 mg, 0.289 mmol), and compound 1-4 is replaced by p-trifluoromethylthiobenzoic acid (54 mg, 0.241 mmol). 52 mg of light yellow solid is obtained, with a yield of 57.23%. Melting point: 182.8-186.1°C. Purity: 99.70%.

[0173] 1 H NMR(300MHz,DMSO-d6)δ12.95(s,1H,NNH),9.11-9.00(m,1H,CONH),7.89(dd,J=46.5,7.9Hz ,4H,Ar-H),7.66(d,J=7.3Hz,3H,Ar-H),7.53-7.30(m,3H,Ar-H),4.50(d,J=4.5Hz,2H,CH2).

[0174] Embodiment 21

[0175] N-((3-phenyl-1H-pyrazol-4-yl)methyl)-3-(trifluoromethyl)benzamide (19)

[0176]

[0177] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (50 mg, 0.289 mmol), and compound 1-4 is replaced by m-trifluoromethylbenzoic acid (46 mg, 0.241 mmol). 46 mg of light yellow solid is obtained, with a yield of 55.31%. Melting point: 119.5-129.3°C. Purity: 98.49%.

[0178] 1 H NMR (300MHz, DMSO-d6) δ13.20-12.78(m,1H,NNH),9.12(t,J=5.3Hz,1H,CONH),8.16(d,J=8.1Hz,2H,Ar-H),7.90(d,J=7. 8Hz,1H,Ar-H),7.79-7.58(m,4H,Ar-H),7.46(t,J=7.5Hz,2H,Ar-H),7.41-7.30(m,1H,Ar-H),4.51(d,J=5.1Hz,2H,CH2).

[0179] Embodiment 22

[0180] N-((3-phenyl-1H-pyrazol-4-yl)methyl)-5,6,7,8-tetrahydronaphthalene-1-carboxamide (20)

[0181]

[0182] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (50 mg, 0.289 mmol), and compound 1-4 is replaced by 5,6,7,8-tetrahydro-1-naphthoic acid (43 mg, 0.241 mmol). 41 mg of light yellow solid is obtained, with a yield of 51.39%. Melting point: 172.7-177.4°C. Purity: 96.59%.

[0183] 1 H NMR(300MHz,DMSO-d6)δ13.15-12.68(m,1H,NNH),8.70-8.47(m,1H,CONH),7.66(d,J=7.6Hz,3H,Ar-H),7.41(dt,J=26.6,7.1Hz,3H,Ar-H), 7.13-7.06(m,2H,Ar-H),7.06-6.98(m,1H,Ar-H),4.43(d,J=5.3Hz,2H,NHCH2),2.80-2.62(m,4H,2×Ar-CH2),1.68(p,J=3.5Hz,4H,(CH2)2).

[0184] Embodiment 23

[0185] 5,6-Difluoro-N-((3-phenyl-1H-pyrazol-4-yl)methyl)-1H-indole-2-carboxamide (21)

[0186]

[0187] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (50 mg, 0.289 mmol), and compound 1-4 is replaced by 5,6-difluoroindole-2-carboxylic acid (48 mg, 0.241 mmol). 45 mg of white solid is obtained, with a yield of 53.04%. Melting point: 206.1-212.8°C. Purity: 98.32%.

[0188] 1 H NMR(300MHz,DMSO-d6)δ13.16-12.79(m,1H,NNH),11.81(s,1H,Indole-NH),9.04-8.83(m,1H,CON H),7.83-7.55(m,4H,Ar-H),7.53-7.28(m,4H,Ar-H),7.19(s,1H,Ar-H),4.64-4.39(m,2H,NHCH2).

[0189] Embodiment 24

[0190] 4-Hexyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (22)

[0191]

[0192] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (50 mg, 0.289 mmol), and compound 1-4 is replaced by p-hexylbenzoic acid (50 mg, 0.241 mmol). 44 mg of white solid is obtained, with a yield of 50.57%. Melting point: 136.1-140.9°C. Purity: 98.38%.

[0193] 1 H NMR(300MHz,DMSO-d6)δ13.15-12.73(m,1H,NNH),8.79(s,1H,CONH),7.78(d, J=7.8Hz,2H,Ar-H),7.67(d,J=7.0Hz,3H,Ar-H),7.52-7.31(m,3H,Ar-H),7.26 (d,J=7.7Hz,2H,Ar-H),4.47(d,J=4.1Hz,2H,CH2),2.61(t,J=7.2Hz,2H,Ar-CH 2),1.65-1.48(m,2H,CH2),1.34-1.20(m,6H,(CH2)3),0.93-0.76(m,3H,CH2).

[0194] Embodiment 25

[0195] 4-Octyl-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (23)

[0196]

[0197] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (50 mg, 0.289 mmol), and compound 1-4 is replaced by p-octylbenzoic acid (57 mg, 0.241 mmol). 54 mg of white solid is obtained, with a yield of 57.60%. Melting point: 143.8-146.1°C. Purity: 97.79%.

[0198] 1 H NMR(300MHz,DMSO-d6)δ13.16-12.71(m,1H,NNH),8.93-8.63(m,1H,CONH),7.78 (d,J=7.8Hz,2H,Ar-H),7.67(d,J=7.6Hz,3H,Ar-H),7.51-7.30(m,3H,Ar-H),7.2 6(d,J=7.9Hz,2H,Ar-H),4.47(d,J=5.2Hz,2H,NHCH2),2.60(t,J=7.6Hz,2H,Ar-C H2),1.65-1.49(m,2H,CH2),1.31-1.18(m,10H,(CH2)5),0.92-0.78(m,3H,CH3).

[0199] Embodiment 26

[0200] 4-(Hexyloxy)-N-((3-phenyl-1H-pyrazol-4-yl)methyl)benzamide (24)

[0201]

[0202] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 3-4 (50 mg, 0.289 mmol), and compound 1-4 is replaced by p-hexyloxybenzoic acid (54 mg, 0.241 mmol). 60 mg of white solid is obtained, with a yield of 66.03%. Melting point: 127.3-131.7°C. Purity: 96.06%.

[0203] 1H NMR(300MHz,DMSO-d6)δ13.20-12.65(m,1H,NNH),8.94-8.44(m,1H,CONH),7.83(d,J= 8.4Hz,2H,Ar-H),7.66(d,J=6.9Hz,3H,Ar-H),7.53-7.41(m,2H,Ar-H),7.40-7.30(m, 1H, Ar-H), 6.96 (d, J = 8.5Hz, 2H, Ar-H), 4.59-4.33 (m, 2H, NHCH2), 4.00 (t, J = 6.2Hz, 2H ,OCH2),1.70(p,J=6.7Hz,2H,CH2),1.46-1.21(m,6H,(CH2)3),0.94-0.80(m,3H,CH3).

[0204] Embodiment 27

[0205] N-((3-(4-Fluorophenyl)-1H-pyrazol-4-yl)methyl)-4-hexylbenzamide (25)

[0206]

[0207] Step 1: Preparation of compound 4-2

[0208] The preparation method is the same as step 1 in Example 4, except that compound 3-1 is replaced by compound 4-1 (2 g, 12.3 mmol), to obtain 1.5 g of a white solid with a yield of 55.93%. 1 H NMR (300MHz, DMSO-d6) δ7.91(s,1H,C=CH),7.78-7.63(m,2H,Ar-H),7.30(t,J=8.9Hz,2H,Ar-H),3.38(s,J=3.3Hz,3H,CH3),3.29(s,3H,CH3).

[0209] Step 2: Preparation of compound 4-3

[0210] The preparation method is the same as step 2 in Example 4, except that compound 3-2 is replaced by compound 4-2 (1.5 g, 6.89 mmol), to obtain 1 g of a white solid with a yield of 77.64%. 1 H NMR (300MHz, DMSO-d6) δ13.93 (s, 1H, NNH), 8.70 (s, 1H, NHCH), 7.91 (dd, J = 8.6, 5.4Hz, 2H, Ar-H), 7.56-7.22 (m, 2H, Ar-H).

[0211] Step 3: Preparation of compound 4-4

[0212] The preparation method is the same as step 3 in Example 4, except that compound 3-3 is replaced by compound 4-3 (500 mg, 2.67 mmol), to obtain 207 mg of a light yellow solid with a yield of 40.59%. 1 H NMR (300MHz, DMSO-d6) δ7.86-7.71 (m, 2H, Ar-H), 7.64 (s, 1H, NHCH), 7.27 (t, J = 8.7Hz, 2H, Ar-H), 3.72 (s, 2H, CH2).

[0213] Step 4: Preparation of compound 25

[0214] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 4-4 (100 mg, 0.523 mmol), and compound 1-4 is replaced by p-hexylbenzoic acid (90 mg, 0.436 mmol). 98 mg of white solid is obtained, with a yield of 59.39%. Melting point: 156.4-158.9°C. Purity: 98.85%.

[0215] 1 H NMR(300MHz,DMSO-d6)δ13.20-12.69(m,1H,NNH),8.79(t,J=5.4Hz,1H,CONH),7.94-7.51(m,5H,Ar-H),7.43-7.10(m,4H,Ar-H),4. 44(d,J=5.3Hz,2H,CH2),2.61(t,J=7.7Hz,2H,CH2),1.56(t,J=7.4Hz,2H,CH2),1.39-1.13(m,6H,(CH2)3),0.97-0.72(m,3H,CH3).

[0216] Embodiment 28

[0217] N-((3-(4-Chlorophenyl)-1H-pyrazol-4-yl)methyl)-4-hexylbenzamide (26)

[0218]

[0219] Step 1: Preparation of compound 5-2

[0220] The preparation method is the same as step 1 in Example 4, except that compound 3-1 is replaced by compound 5-1 (2 g, 11.2 mmol), to obtain 1.5 g of a white solid with a yield of 57.23%. 1H NMR (300MHz, DMSO-d6) δ7.90(s,1H,C=CH),7.63(d,J=8.2Hz,2H,Ar-H),7.53(d,J=8.2Hz,2H,Ar-H),3.37(s,3H,CH3),3.29(s,3H,CH3).

[0221] Step 2: Preparation of compound 5-3

[0222] The preparation method is the same as step 2 in Example 4, except that compound 3-2 is replaced by compound 5-2 (1.5 g, 6.89 mmol). 1 g of white solid is obtained, with a yield of 76.91%. 1 H NMR (300MHz, DMSO-d6) δ 14.41-13.74 (m, 1H, NNH), 8.74 (s, 1H, NHCH), 7.91 (d, J = 8.3Hz, 2H, Ar-H), 7.61 (d, J = 8.2Hz, 2H, Ar-H).

[0223] Step 3: Preparation of compound 5-4

[0224] The preparation method is the same as step 3 in Example 4, except that compound 3-3 is replaced by compound 5-3 (500 mg, 2.46 mmol), to obtain 210 mg of a light yellow solid with a yield of 41.25%. 1 H NMR (300MHz, DMSO-d6) δ7.81-7.60(m,3H,Ar-H,NNH),7.51-7.39(m,2H,Ar-H),7.39 -7.27(m,1H,Ar-H),3.76(s,2H,CH2).

[0225] Step 4: Preparation of compound 26

[0226] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 5-4 (100 mg, 0.483 mmol), and compound 1-4 is replaced by p-hexylbenzoic acid (83 mg, 0.402 mmol). 100 mg of white solid is obtained, with a yield of 62.94%. Melting point: 130.3-133.0°C. Purity: 99.11%.

[0227] 1H NMR(300MHz,DMSO-d6)δ13.24-12.66(m,1H,NNH),8.80(t,J=5.4Hz,1H,CONH),7.78(d ,J=8.0Hz,2H,Ar-H),7.74-7.60(m,2H,Ar-H),7.56-7.41(m,2H,Ar-H),7.37(d,J=7.4 Hz,1H,Ar-H),7.26(d,J=7.9Hz,2H,Ar-H),4.47(d,J=5.3Hz,2H,CH2),2.61(t,J=7.6H z,2H,CH2),1.71-1.43(m,2H,CH2),1.41-1.11(m,6H,(CH2)3),0.95-0.75(m,3H,CH3).

[0228] Embodiment 29

[0229] N-((3-(3-Fluorophenyl)-1H-pyrazol-4-yl)methyl)-4-hexylbenzamide (27)

[0230]

[0231] Step 1: Preparation of compound 6-2

[0232] The preparation method is the same as step 1 in Example 4, except that compound 3-1 is replaced by compound 6-1 (1.3 g, 7.99 mmol), to obtain 0.9 g of a white solid with a yield of 51.66%. 1 H NMR (300MHz, Chloroform-d) δ7.96 (s, 1H, C=CH), 7.58 (d, J=7.7Hz, 1H, Ar-H), 7.5 1-7.34(m,2H,Ar-H),7.23-7.10(m,1H,Ar-H),3.50(s,3H,CH3),3.31(s,3H,CH3).

[0233] Step 2: Preparation of compound 6-3

[0234] The preparation method is the same as step 2 in Example 4, except that compound 3-2 is replaced by compound 6-2 (0.9 g, 4.13 mmol), to obtain 541 mg of a white solid with a yield of 70.07%. 1 H NMR (300MHz, DMSO-d6) δ14.05 (s, 1H, NNH), 8.69 (s, 1H, NHCH), 7.75 (d, J = 7.7Hz, 1H, Ar-H), 7.70-7.55 (m, 2H, Ar-H), 7.40-7.21 (m, 1H, Ar-H).

[0235] Step 3: Preparation of compound 6-4

[0236] The preparation method is the same as step 3 in Example 4, except that compound 3-3 is replaced by compound 6-3 (541 mg, 2.89 mmol), to obtain 183 mg of a light yellow solid with a yield of 33.11%. 1 H NMR (300MHz, DMSO-d6) δ7.77(s,1H,NHCH),7.60-7.44(m,3H,Ar-H),7.26-7.13(m,1H,Ar-H),4.56(brs,2H,NH2),3.84(s,2H,CH2),3.44(brs,1H,NNH).

[0237] Step 4: Preparation of compound 27

[0238] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 6-4 (100 mg, 0.523 mmol), and compound 1-4 is replaced by p-hexylbenzoic acid (90 mg, 0.436 mmol). 106 mg of white solid is obtained, with a yield of 64.23%. Melting point: 135.4-138.2°C. Purity: 99.04%.

[0239] 1 H NMR (300MHz, DMSO-d6) δ13.29-12.77(m,1H,NNH),8.75(s,1H,CONH),7.77(d,J=7.7Hz,3H,Ar-H),7.51(dd,J=15.0,9.2Hz,3H,Ar-H),7.26(d,J=7.6Hz,2H, Ar-H),7.17(s,1H,Ar-H),4.48(d,J=6.1Hz,2H,CH2),2.61(t,J=7.6Hz,2H,CH 2),1.66-1.48(m,2H,CH2),1.37-1.16(m,6H,(CH2)3),0.93-0.72(m,3H,CH3).

[0240] Embodiment 30

[0241] N-((3-(3-Chlorophenyl)-1H-pyrazol-4-yl)methyl)-4-hexylbenzamide (28)

[0242]

[0243] Step 1: Preparation of compound 7-2

[0244] The preparation method is the same as step 1 in Example 4, except that compound 3-1 is replaced by compound 7-1 (2 g, 11.2 mmol), to obtain 1.5 g of a white solid with a yield of 57.23%. 1 H NMR (300MHz, Chloroform-d) δ7.96 (s, 1H, C=CH), 7.75-7.64 (m, 2H, Ar-H), 7.46 (ddd, J=8. 0, 2.1, 1.2Hz, 1H, Ar-H), 7.36 (t, J = 7.8Hz, 1H, Ar-H), 3.50 (s, 3H, CH3), 3.32 (s, 3H, CH3).

[0245] Step 2: Preparation of compound 7-3

[0246] The preparation method is the same as step 2 in Example 4, except that compound 3-2 is replaced by compound 7-2 (1.5 g, 6.89 mmol), to obtain 1 g of a white solid with a yield of 76.91%. 1 H NMR (300MHz, DMSO-d6) δ14.06 (s, 1H, NNH), 8.74 (s, 1H, NHCH), 7.90 (t, J = 1.8Hz, 1H, Ar-H), 7.88-7.80 (m, 1H, Ar-H), 7.65-7.51 (m, 2H, Ar-H).

[0247] Step 3: Preparation of compound 7-4

[0248] The preparation method is the same as step 3 in Example 4, except that compound 3-3 is replaced by compound 7-3 (500 mg, 2.46 mmol), to obtain 176 mg of a light yellow solid with a yield of 34.57%. 1 H NMR (300MHz, DMSO-d6) δ7.76-7.64(m,2H,Ar-H),7.54-7.21(m,3H,Ar-H),3.78(s,2H,CH2).

[0249] Step 4: Preparation of compound 28

[0250] The preparation method is the same as step 2 in Example 2, except that p-butylaniline is replaced by compound 7-4 (70 mg, 0.338 mmol), and compound 1-4 is replaced by p-hexylbenzoic acid (55 mg, 0.282 mmol). 46 mg of white solid is obtained, with a yield of 41.29%. Melting point: 135.9-138.9°C. Purity: 95.06%.

[0251] 1H NMR(500MHz,DMSO-d6)δ13.17-12.59(m,1H,NNH),8.80-8.64(m,1H,CONH),7.77(d, J=7.8Hz,2H,Ar-H),7.66(d,J=7.7Hz,2H,Ar-H),7.52-7.40(m,2H,Ar-H),7.40-7.3 1(m,1H,Ar-H),7.25(d,J=7.8Hz,2H,Ar-H),4.47(d,J=5.3Hz,2H,CH2),2.61(t,J=7 .7Hz,2H,CH2),1.68-1.43(m,2H,CH2),1.27(s,6H,(CH2)3),0.97-0.70(m,3H,CH3).

[0252] Example 31 Biological Activity

[0253] The inhibitory effect of the compound on the cGAMP-stimulated STING pathway in THP1-Dual cells was detected by the luciferase method as follows:

[0254] (1) 4*10 THP1-Dual cells in the logarithmic growth phase were cultured. 4 Each well was seeded in a 96-well plate, and compounds of different concentrations were added, and the DMSO group was used as a blank control.

[0255] (2) Cells were transfected with 10 μg / mL cGAMP and lipofectamine 2000 (Invitrogen) and cultured for 24 h.

[0256] (3) Using QUANTI-Luc TM 10 μL of cell culture supernatant was added to a 96-well plate, and then 50 μL of QUANTI-Luc TM The reagents were read using a multifunctional microplate reader (Thermo).

[0257] Relative luciferase activity calculation method: cells treated with lipofectamine 2000 were used as blank control, and cells treated with lipofectamine 2000:cGAMP complex were used as negative control.

[0258] Relative luciferase activity = (RLU - RLU blackcontrol) / (RLU negativecontrol - RLU blackcontrol), RLU represents the original luciferase value.

[0259] Measured compound IC 50, and compared with the positive control drug H151 as shown in Table 1:

[0260] Table 1 Compound IC 50

[0261]

[0262] It can be seen from the above table that the above compounds of the present invention have a good inhibitory effect on the cGAS-STING pathway, can provide a basis for the preparation of drugs for treating diseases related to the function of the STING protein, have the prospect of preparing drugs for treating and / or preventing inflammatory diseases and autoimmune diseases, and provide a reliable approach for clinical application.

[0263] Example 32 Evaluation of the binding ability of compounds 5 and 22 to STING by isothermal titration calorimetry (ITC)

[0264] The ITC experiment uses the MicroCal ITC 200 instrument to measure the binding affinity between proteins and ligands. The ITC experiment can fit and quantitatively calculate the binding thermodynamic parameters, including the number of binding sites (n), binding constant (Ka), reaction enthalpy change (ΔH), entropy change (ΔS), etc., by detecting the heat change (endothermic or exothermic) when the protein solution and the small molecule (ligand) solution interact.

[0265] High-concentration, high-purity STING-CTD protein was obtained through extraction, dialysis, and ultrafiltration. During the dialysis process, the buffer consistent with the protein component system was retained as much as possible. First, 400 μL STING-CTD protein (1 μM) and 100 μL compound (10 μM) were prepared, and the DMSO content in the protein and compound needed to be consistent. Then, an isothermal titration calorimeter (MicroCal iTC200) was used, and the operating parameters were set: temperature 25 °C; speed 750 rpm. The sample pool and titration needle were cleaned, and about 200 μL of STING-CTD protein was slowly added to the sample pool. About 50 μL of the compound was inhaled into the titration needle, and 2 μL was dripped into the sample pool at a drop interval of 2.5 min per drop, for a total of 19 drops. After the experiment, MicroCal.Origin version 7.0 software was used for data processing and analysis to obtain the entropy change ΔS, enthalpy change ΔH and free energy ΔG change of the compound binding to the STING-CTD protein, and then the KA value and KD value (KD = 1 / KA) were obtained. The smaller the KD value, the better the binding strength and selectivity of the compound to the protein.

[0266] The compound 5 tested showed binding activity to STING. Figure 1 As shown, the binding affinity constant KD of compound 5 is 53 nM.

[0267] The compound 22 tested showed binding to STING. Figure 2 As shown, the binding affinity constant KD of compound 22 is 163 nM.

[0268] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the present invention itself. Various changes in form and details may be made without departing from the spirit and scope of the present invention as defined in the appended claims.

Claims

1. A 3-phenyl-1H-pyrazole compound, characterized in that Selected from:

2. The compound according to claim 1, characterized in that The pharmaceutically acceptable salts include acid addition salts formed between the compound of claim 1 and the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, maleic acid, benzenesulfonic acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, fumaric acid, salicylic acid or phenylacetic acid; and also include acid salts formed between the compound of claim 1 and inorganic bases or organic salts made from basic amines.

3. A pharmaceutical composition, characterized in that It comprises the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

4. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating a disease related to the function of the STING protein.

5. The use according to claim 4, characterized in that The diseases associated with STING protein function include inflammatory diseases and autoimmune diseases.

6. The use according to claim 4, characterized in that The diseases related to the function of STING protein include STING-related infantile-onset vasculopathy SAVI, Aicardi-Goutières syndrome AGS, COPA syndrome caused by gene mutation of the α subunit of the eutectic protein complex, systemic lupus erythematosus SLE, familial pernio lupus erythematosus FCL, Parkinson's disease PD, amyotrophic lateral sclerosis ALS, Huntington's disease, non-alcoholic steatohepatitis NASH, alcoholic liver, nerve damage, rheumatoid arthritis, renal fibrosis, systemic sclerosis, intervertebral disc degeneration, pulmonary fibrosis, aging, scleroderma, psoriasis, inflammatory bowel disease, autoimmune colitis, irritable bowel syndrome, ulcerative colitis, Crohn's disease, uveitis, mucositis, diabetes, cardiovascular disease or neurodegenerative disease.

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