A 2-Benzofurancarboxylic Acid Derivative, Its Preparation Method and Use

Abstract

The present invention discloses a 2-benzofuran carboxylic acid derivative represented by general formula I or its enantiomer, diastereomer, racemate, pharmaceutically acceptable salt, and a pharmaceutical composition containing the same. Such compounds, as cGAS-STING pathway targeting inhibitors, can target and inhibit the cGAMP-activated cGAS-STING pathway, and have the use for preparing drugs for treating and / or preventing inflammatory diseases and autoimmune diseases.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to a 2-benzofuran carboxylic acid derivative, a preparation method and a use thereof. Such compounds can be used as cGAS-STING pathway targeting inhibitors. Background Art

[0002] The first line of defense of the mammalian innate immune system relies on pattern recognition receptors (PRRs) to detect extracellular and intracellular danger signals. Ligand-bound PRRs can trigger the transcription of inflammatory genes and induce the expression of related cytokines, thereby promoting adaptive immune activation to achieve the purpose of eradicating pathogens and repairing cell damage. Currently known PRRs can be divided into five categories: retinoic acid-induced gene I-like receptors (RLRs), Toll-like receptors (TLRs), C-type lectin receptors (CLRs), nucleotide binding oligomeric domain-like receptors (NLRs) and cytosolic DNA sensors. Recently, cyclic guanosine monophosphate-adenosine (cGAMP) synthase (cGAS) has been identified as a cytosolic DNA sensor. After cGAS recognizes abnormal DNA molecules from bacteria or itself, it can initiate the cGAS-stimulator of interferon genes (STING) signaling pathway to induce an immune response. As a key signal adapter connecting the cGAS receptor and downstream effectors, STING is localized in the endoplasmic reticulum, where it recognizes cGAMP produced by cGAS and stimulates the production of interferon-β (IFN-β) and other pro-inflammatory factors.

[0003] STING (Stimulator of interferon genes, also known as TMEM173, ERIS, MITA, and MPYS), since its discovery in 2008, continuous exploration of its pathway has led to a deeper understanding of its functions and potential therapeutic applications. Pharmaceutical researchers have invested in developing cancer treatments using STING agonists. However, increasing evidence suggests that the abnormal activation of the cGAS-STING pathway plays a functional role in inflammatory and autoimmune diseases. Chronic activation of the STING pathway may lead to autoimmune diseases such as systemic lupus erythematosus (SLE), Aicardi-Goutières syndrome (AGS), and STING-associated vasculopathy with onset in infancy (SAVI). The pathogenesis of these types of diseases may be attributed to mutations in nucleases and STING proteins. When self-DNA enters the cytoplasm and cannot be effectively degraded by defective DNA enzymes (such as TREX1 mutants), it is detected by cGAS and produces cGAMP. This triggers abnormal activation of the STING pathway accompanied by overproduction of chemokines and cytokines, leading to tissue damage (SLE and AGS). Similarly, gene mutations can induce abnormal activation of the STING protein, and the STING pathway remains persistently activated (SAVI) even without the stimulation of CDN ligands. Therefore, STING-targeted inhibitors are an attractive approach for treating autoinflammatory diseases.

[0004] It can be seen that the cGAS-STING pathway plays an important role in the process of autoinflammation and autoimmune diseases. Inhibiting the activation of this pathway can slow down the progression of the disease. The study of the STING signaling pathway may be a new direction for the treatment of autoimmune diseases. Summary of the Invention

[0005] Object of the Invention: One object of the present invention is to provide a 2-benzofuran carboxylic acid derivative of general formula I or its enantiomers, diastereomers, racemates, pharmaceutically acceptable salts:

[0006]

[0007] Wherein,

[0008] A is selected from H, substituted or unsubstituted alkyl, substituted or unsubstituted 3-8 membered cycloalkyl or heterocyclic group, substituted or unsubstituted 5-8 membered heteroaryl or aryl;

[0009] B is selected from substituted or unsubstituted C 1 -C 2 alkyl acyl, substituted or unsubstituted amino acyl, substituted or unsubstituted C 1 -C 2an alkylamide group, a substituted or unsubstituted 3- to 8-membered cycloalkyl or heterocyclic group, a substituted or unsubstituted 5- to 8-membered heteroaryl or aryl group;

[0010] L C selected from H, -NHCO-, -NHSO 2 - or -NH-;

[0011] D is selected from H, a substituted or unsubstituted C 1 -C 6 alkyl or heteroalkyl group, a substituted or unsubstituted C 2 -C 6 alkenyl group, a substituted or unsubstituted C 2 -C 6 alkynyl group, a substituted or unsubstituted 3- to 8-membered heterocyclic or cycloalkyl group, a substituted or unsubstituted 5- to 8-membered heteroaryl or aryl group.

[0012] In certain preferred embodiments,

[0013] A is selected from H, a substituted or unsubstituted alkyl group;

[0014] B is selected from a substituted or unsubstituted 5- to 8-membered heteroaryl or aryl group;

[0015] L C is selected from -NHCO- or -NH-;

[0016] D is selected from a substituted or unsubstituted 5- to 8-membered heteroaryl or aryl group.

[0017] In certain further preferred embodiments,

[0018] A is selected from difluoromethyl, trifluoromethyl or ethyl;

[0019] B is selected from a benzene ring, a pyrazole ring, an imidazole ring, an oxazole ring or a pyrrole ring;

[0020] D is selected from a benzene ring substituted with a tert-butyl group.

[0021] In certain preferred embodiments,

[0022] A is selected from a substituted or unsubstituted 5- to 8-membered heteroaryl or aryl group;

[0023] B is selected from a substituted or unsubstituted 5- to 8-membered heteroaryl or aryl group;

[0024] L C is selected from H, -NHCO- or -NH-;

[0025] D is selected from H, a substituted or unsubstituted C 1 -C 6 alkyl or heteroalkyl group, a substituted or unsubstituted 3- to 8-membered heterocyclic or cycloalkyl group, a substituted or unsubstituted 5- to 8-membered heteroaryl or aryl group.

[0026] In certain further preferred embodiments,

[0027] A is selected from a benzene ring mono- or disubstituted with ortho- or para-methyl, ethyl, isopropyl, tert-butyl, halogen, n-propyl, methoxy, cyano, trifluoromethyl, or trifluoromethoxy;

[0028] B is selected from a pyrazole ring;

[0029] D is selected from ethyl, cyclohexyl, or a benzene ring substituted with p-tert-butyl.

[0030] In some preferred embodiments, the pharmaceutically acceptable salts refer to non-toxic inorganic or organic acid and / or base addition salts. See, for example, Lit et al., Salt Selection for Basic Drugs (1986), Int J. Pharm., 33, 201-217, which is incorporated herein by reference.

[0031] The compounds of general formula I of the present invention are preferably the following compounds:

[0032]

[0033]

[0034] The above-mentioned compounds of general formula I of the present invention may also exist in the form of their salts, which are converted into compounds of general formula I in vivo. For example, within the scope of the present invention, according to the processes known in the art, the compounds of the present invention are converted into the form of pharmaceutically acceptable salts and used in the form of salts.

[0035] All tautomeric forms of the compounds of 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 stereoisomeric forms. Additional asymmetric carbon atoms may exist in substituents such as alkyl, and all such isomers and their mixtures are included within the scope of the present invention.

[0036] The present invention also provides a method for preparing the compounds of general formula I, and the synthetic route is as follows:

[0037]

[0038] Reagents and conditions: (a) Copper acetate anhydrous, pyridine, dichloromethane, room temperature; (b) Bis(pinacolato)diboron, Pd(dppf)Cl 2 , potassium acetate, N 2 , 80 °C; (c) Methyl bromoacetate, potassium carbonate, N,N-dimethylacetamide, 105 °C; (d) Di-tert-butyl dicarbonate, triethylamine, 4-dimethylaminopyridine, dichloromethane, 40 °C; (e) Sphos Pd G3 , cesium carbonate, water + 1,4-dioxane (1:10, v / v), N 2 , 80 °C; (f) trifluoroacetic acid, dichloromethane, room temperature; (g) anhydrous pyridine, room temperature; (h) aqueous lithium hydroxide solution, room temperature.

[0039] wherein, A, B, L C 、D are as defined in General Formula I.

[0040] Another object of the present invention is to provide a pharmaceutical composition, the pharmaceutical composition comprising a compound of General Formula I or a pharmaceutically acceptable salt thereof, its enantiomers, diastereomers, racemates, and a pharmaceutically acceptable carrier.

[0041] The pharmaceutical compositions 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 compositions 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, and suspensions, dispersions, and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, binders, colorants, fillers, emulsifiers, etc.

[0042] Sterile injectable compositions can be formulated using suitable dispersing 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, etc.

[0043] Topical compositions can be formulated as oils, lotions, creams, etc. Carriers for the compositions include vegetable oils or mineral oils, animal fats, and high molecular weight alcohols, etc. A pharmaceutically acceptable carrier is a carrier in which the active ingredient is soluble.

[0044] The actual dosage level of the active ingredient in the pharmaceutical compositions of the present invention can be varied so as to obtain an amount of the active ingredient that is effective in achieving the desired therapeutic response for a particular patient, composition, and mode of administration and that is non-toxic to the patient. The selected dosage level depends on a variety of factors, including the activity of the specific compound or salt of the present invention 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 status, and medical history of the patient being treated, and similar factors well known in the medical arts.

[0045] Another object of the present invention is to provide the use of a compound of general formula I or a pharmaceutically acceptable salt thereof, its enantiomers, diastereomers, and racemates in the preparation of a drug for preventing and / or treating inflammatory diseases and autoimmune diseases.

[0046] The inflammatory diseases and autoimmune diseases are selected from systemic lupus erythematosus (SLE), familial chilblain lupus erythematosus (FCL), Aicardi-Goutières syndrome (AGS), STING-associated vasculopathy with onset in infancy (SAVI), Singleton-Merten syndrome (SMS), amyotrophic lateral sclerosis (ALS), Parkinson's syndrome (PD), non-alcoholic steatohepatitis (NASH), pulmonary fibrosis, aging, scleroderma, psoriasis, rheumatoid arthritis, inflammatory bowel disease, autoimmune colitis, irritable bowel syndrome, ulcerative colitis, Crohn's disease, uveitis, mucositis, diabetes, cardiovascular diseases, and neurodegenerative diseases.

[0047] The compound of general formula I prepared by the present invention and its pharmaceutically acceptable salt, its enantiomers, diastereomers, and racemates, as cGAS-STING pathway targeting inhibitors, can target and inhibit the cGAMP-activated cGAS-STING pathway, and have the prospect of preparing drugs for treating and / or preventing inflammatory diseases and autoimmune diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 It is a graph of ITC titration of compound 8 with STING protein. DETAILED DESCRIPTION OF THE INVENTION

[0049] The preparation method of the compound of general formula I of the present invention will be described below in conjunction with specific examples, but these specific methods do not constitute any limitation to the present invention. The compounds 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 combinations can be easily carried out by those skilled in the art to which the present invention pertains.

[0050] The starting materials, reaction reagents, etc. used in the specific examples of the present invention are all commercially available. The present invention can be prepared in the form of salts by using common salt-forming methods in the art. For example, at room temperature, the compound is dissolved in hydrochloric acid ethanol for reaction to form hydrochloride; or benzenesulfonic acid is added thereto for reaction to form benzenesulfonate.

[0051] The experimental methods without specific conditions noted in the examples of the present invention are usually carried out under conventional conditions or according to the conditions recommended by the raw material or commodity manufacturers.

[0052] The structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR shift (δ) is in 10-6 Given in the unit of (ppm). The NMR measurement was performed using a Bruker AVANCE-400 nuclear magnetic resonance spectrometer, and the solvent for the measurement was deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), and the internal standard was tetramethylsilane (TMS).

[0053] The starting materials known in the present invention can be adopted or synthesized according to methods known in the art, or can be purchased from companies such as ABCR GmbH&Co.KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, Darui Chemicals, etc.

[0054] Unless otherwise specified in the examples, the reactions were carried out under an air atmosphere.

[0055] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0056] Unless otherwise specified in the examples, the reaction temperature was room temperature, which was 20 - 30 °C.

[0057] In the examples, the progress of the reaction was monitored by thin-layer chromatography (TLC). The eluent system for column chromatography and the developing agent system for thin-layer chromatography included: A: petroleum ether / ethyl acetate system, B: dichloromethane / methanol system. The volume ratio of the solvents was adjusted according to the polarity of the compound, and a small amount of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0058] The thin-layer chromatography silica gel plate used was Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the TLC silica gel plate was 0.15 mm - 0.2 mm, and the specification used for separating and purifying products by thin-layer chromatography was 0.4 mm - 0.5 mm.

[0059] For silica gel column chromatography, silica gel with 200 - 300 mesh from Yantai Huanghai was generally used as the carrier.

[0060] Example 1: Synthesis of 3-(4-(tert-butyl)benzamido)-5-(1-(difluoromethyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 1)

[0061]

[0062] 1. Synthesis of Compound 1-2

[0063] Dissolve 5-bromo-2-hydroxybenzonitrile (10 g, 65.12 mmol, 1 eq) in 150 mL of DMF, add methyl bromoacetate (10.96 g, 71.63 mmol, 1.1 eq), potassium carbonate (18 g, 130.24 mmol, 2 eq), heat to 105 °C, after reacting for 8 h, cool naturally, add 150 mL of water, stir at room temperature for 30 minutes, filter to obtain a pale yellow solid, slurry with petroleum ether:ethyl acetate (volume ratio 10:1) to obtain compound 1-2 (10.76 g, yield: 60.85%). 1 H NMR(300MHz,DMSO-d 6 )δ8.22(d,J=2.1Hz,1H),7.62(dd,J=8.8,2.1Hz,1H),7.48(d,J=8.8Hz,1H),6.42(s,2H),3.81(s,3H)。

[0064] 2. Synthesis of compound 1-3

[0065] Dissolve compound 1-2 (3.29 g, 12.28 mmol, 1 eq) in 30 mL of DCM, add 4-dimethylaminopyridine (300 mg, 2.45 mmol, 0.2 eq), di-tert-butyl dicarbonate (3.39 ml, 14.73 mmol, 1.2 eq), triethylamine (2.56 ml, 18.42 mmol, 1.5 eq), heat to 40 °C, after reacting for 4 h, distill under reduced pressure, make sand and pass through a column to obtain compound 1-3 (2.746 g, yield: 60.6%). 1 H NMR(300MHz,CDCl 3 )δ8.65(d,J=2.1Hz,1H),8.56(s,1H),7.60(dd,J=8.9,2.1Hz,1H),7.39(d,J=8.9Hz,1H),4.05(s,3H),1.60(s,9H)。

[0066] 3. Synthesis of compound 1-5

[0067] Dissolve compound 1-3 (144 mg, 0.39 mmol, 1 eq) and compound 1-4 (107.4 mg, 0.44 mmol, 1.12 eq) in a mixed solution of water and 1,4-dioxane (volume ratio 1:10) (10 ml), add cesium carbonate (306.3 mg, 0.94 mmol, 2.4 eq), Sphos Pd G 3 (23.4mg,0.03mmol,0.08eq)N 2Protect, heat up to 80 °C, after reacting for 6 h, remove the solvent by vacuum distillation, make sand and pass through a column to obtain Compound 1-5 (151.74 mg, yield: 95.57%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.23 (s, 1H), 8.76 (s, 1H), 8.29 (s, 1H), 8.15 - 8.05 (m, 1H), 7.93 - 7.84 (m, 1H), 7.77 - 7.65 (m, 1H), 3.88 (s, 3H), 1.51 (s, 9H).

[0068] 4. Synthesis of Compound 1-6

[0069] Dissolve Compound 1-5 in a dichloromethane solution of trifluoroacetic acid (volume ratio 1:5, 10 ml), stir at room temperature for 6 h, add saturated sodium bicarbonate aqueous solution dropwise to adjust the solution to alkaline, extract with dichloromethane three times, combine the organic phases, dry over anhydrous sodium sulfate, filter, remove the solvent by vacuum distillation, make sand and pass through a column to obtain Compound 1-6 (100 mg, 87.37%). 1 HNMR (300 MHz, DMSO-d 6 ) δ 8.64 (s, 1H), 8.29 (d, J = 1.8 Hz, 1H), 8.20 (s, 1H), 7.93 - 7.78 (m, 2H), 7.54 (d, J = 8.7 Hz, 1H), 6.38 (s, 2H), 3.83 (s, 3H).

[0070] 5. Synthesis of Compound 1-7

[0071] Dissolve Compound 1-6 (100 mg, 0.33 mmol, 1 eq) in 5 ml of anhydrous pyridine, slowly add p-tert-butylbenzoyl chloride (66 μL, 0.33 mmol, 1 eq) dropwise at 0 °C. After the addition is complete, stir at room temperature for 1 h, add dilute hydrochloric acid to neutralize pyridine, extract with dichloromethane three times, combine the organic phases, dry over anhydrous sodium sulfate, filter, remove the solvent by vacuum distillation, make sand and pass through a column to obtain Compound 1-7 (149.54 mg, yield: 97%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.40 (s, 1H), 8.81 (s, 1H), 8.34 (s, 1H), 8.16 (s, 1H), 8.05 (d, J = 7.6 Hz, 2H), 7.98 - 7.75 (m, 2H), 7.64 (d, J = 8.7 Hz, 2H), 3.91 (s, 3H), 1.37 (s, 9H).

[0072] 6. Synthesis of Compound 1

[0073] Compound 1-7 (79.4 mg, 0.2 mmol) was dissolved in THF:H 2 O (volume ratio 1:1, 5 ml), 1 M aqueous LiOH solution was added, and the reaction was carried out at room temperature for 2 h. The reaction was monitored by TLC. After the reaction was completed, the pH was adjusted to 1 with 1 M aqueous HCl solution, filtered, and dried to obtain Compound 1. (White solid, 85.99 mg, yield: 94.88%) 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.44 (s, 1H), 8.78 (s, 1H), 8.30 (s, 1H), 8.17 (d, J = 1.8 Hz, 1H), 8.02 (dd, J = 8.4, 1.7 Hz, 2H), 7.93 - 7.81 (m, 1H), 7.75 (d, J = 8.8 Hz, 1H), 7.68 - 7.57 (m, 2H), 1.34 (s, 9H).

[0074] Example 2: Synthesis of 3-(4-(tert-butyl)benzamido)-5-(1-ethyl-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 2)

[0075]

[0076] The preparation method was the same as that of Example 1, except that Compound 2-4 was replaced with Compound 1-4.

[0077] Compound 2-5 (351 mg, yield: 97.63%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.19 (s, 1H), 8.31 - 8.12 (m, 1H), 7.99 (d, J = 1.8 Hz, 1H), 7.91 - 7.84 (m, 1H), 7.76 (dd, J = 8.7, 1.9 Hz, 1H), 7.66 (d, J = 8.7 Hz, 1H), 4.19 (q, J = 7.3 Hz, 2H), 3.88 (s, 3H), 1.51 (s, 9H), 1.42 (t, J = 7.2 Hz, 3H).

[0078] Compound 2-6 (225.19 mg, yield: 86.67%) was obtained. 1 H NMR (300 MHz, DMSO-d 6)δ 8.20 - 8.09 (m, 2H), 7.85 - 7.78 (m, 1H), 7.70 (dd, J = 8.7, 1.9 Hz, 1H), 7.47 (d, J = 8.6 Hz, 1H), 6.35 (s, 2H), 4.19 (q, J = 7.3 Hz, 2H), 3.83 (s, 3H), 1.44 (t, J = 7.3 Hz, 3H).

[0079] Compound 2 - 7 (330.99 mg, yield: 94.13%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 )δ 10.39 (s, 1H), 8.28 - 8.22 (m, 1H), 8.08 - 7.96 (m, 3H), 7.93 - 7.87 (m, 1H), 7.81 (dd, J = 8.8, 1.8 Hz, 1H), 7.72 (d, J = 8.8 Hz, 1H), 7.63 (d, J = 8.4 Hz, 2H), 4.16 (q, J = 7.3 Hz, 2H), 3.89 (s, 3H), 1.41 (t, J = 7.3 Hz, 3H), 1.36 (s, 9H).

[0080] Compound 2 (white solid, 314.93 mg, yield: 98.24%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 )δ 13.80 (s, 1H), 10.35 (s, 1H), 8.24 (s, 1H), 8.08 - 7.99 (m, 3H), 7.92 - 7.86 (m, 1H), 7.79 (dd, J = 8.7, 1.8 Hz, 1H), 7.71 (d, J = 8.7 Hz, 1H), 7.63 (d, J = 8.4 Hz, 2H), 4.17 (q, J = 7.3 Hz, 2H), 1.42 (t, J = 7.3 Hz, 3H), 1.36 (s, 9H).

[0081] Example 3: Synthesis of 3-(4-(tert - butyl)benzamido)-5-(4-(trifluoromethyl)phenyl)benzofuran - 2 - carboxylic acid (Compound 3)

[0082]

[0083] The preparation method was the same as that of Example 1, except that Compound 3 - 4 was replaced with Compound 1 - 4.

[0084] Compound 3 - 5 (241.45 mg, yield: 97.13%) was obtained. 1 H NMR (300 MHz, CDCl 3) δ 7.78 - 7.75 (m, 1H), 7.73 (s, 4H), 7.69 (d, J = 1.9 Hz, 1H), 7.61 - 7.51 (m, 1H), 4.01 (s, 3H).

[0085] Compound 3 - 6 (147.62 mg, yield: 97.65%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 8.04 (d, J = 1.8 Hz, 1H), 8.01 (d, J = 2.6 Hz, 1H), 7.99 (s, 1H), 7.98 (d, J = 1.9 Hz, 1H), 7.92 (d, J = 8.8 Hz, 1H), 7.85 (d, J = 8.2 Hz, 2H), 3.93 (s, 3H), 1.38 (s, 18H).

[0086] Compound 3 - 7 (215.09 mg, yield: 98.6%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 10.46 (s, 1H), 8.27 (d, J = 1.9 Hz, 1H), 8.02 (d, J = 8.3 Hz, 2H), 7.99 - 7.80 (m, 6H), 7.63 (d, J = 8.2 Hz, 2H), 3.92 (s, 3H), 1.35 (s, 10H).

[0087] Compound 3 (white solid, 203.53 mg, yield: 97.38%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 10.68 (s, 1H), 8.36 (s, 1H), 8.05 - 7.97 (m, 2H), 7.92 (s, 4H), 7.85 (s, 2H), 7.66 - 7.57 (m, 2H), 1.34 (s, 9H).

[0088] Example 4: Synthesis of 3 - (4 - (tert - butyl)benzamido) - 5 - (4 - ethylphenyl)benzofuran - 2 - carboxylic acid (Compound 4)

[0089]

[0090] The preparation method was the same as that of Example 1, except that Compound 4 - 4 was replaced with Compound 1 - 4.

[0091] Compound 4 - 5 (201 mg, yield: 89.3%) was obtained. 1H NMR (300 MHz, CDCl 3) δ 8.40 (s, 1H), 7.85 (s, 1H), 7.69 (s, 1H), 7.48 - 7.42 (m, 2H), 7.24 - 7.18 (m, 2H), 3.90 (s, 2H), 2.68 (t, J = 1.0 Hz, 2H), 1.26 (s, 2H).

[0092] Compound 4 - 6 (119.78 mg, yield: 96.54%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 8.29 (d, J = 1.9 Hz, 1H), 7.78 (dd, J = 8.7, 2.0 Hz, 1H), 7.66 - 7.58 (m, 2H), 7.55 (d, J = 8.7 Hz, 1H), 7.34 (d, J = 8.1 Hz, 2H), 6.45 (s, 2H), 3.83 (s, 3H), 2.67 (q, J = 7.6 Hz, 2H), 1.23 (t, J = 7.6 Hz, 3H).

[0093] Compound 4 - 7 (173.67 mg, yield: 94%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 10.44 (s, 1H), 8.14 (d, J = 1.8 Hz, 1H), 8.02 (d, J = 8.4 Hz, 2H), 7.93 - 7.76 (m, 2H), 7.62 (dd, J = 8.2, 5.4 Hz, 4H), 7.33 (d, J = 7.9 Hz, 2H), 3.91 (s, 3H), 2.66 (q, J = 7.6 Hz, 2H), 1.36 (s, 9H), 1.22 (t, J = 7.6 Hz, 3H).

[0094] Compound 4 (164.62 mg, yield: 97.8%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 10.44 (s, 1H), 8.17 (dd, J = 16.3, 1.8 Hz, 1H), 8.03 (dd, J = 8.5, 1.8 Hz, 2H), 7.89 - 7.74 (m, 2H), 7.68 - 7.57 (m, 4H), 7.34 (d, J = 7.9 Hz, 2H), 2.67 (q, J = 7.6 Hz, 2H), 1.36 (d, J = 1.5 Hz, 9H), 1.23 (t, J = 7.5 Hz, 4H).

[0095] Example 5: Synthesis of 3 - (4 - (tert - butyl)benzamido) - 5 - (1 - (4 - ethylphenyl) - 1H - pyrazol - 4 - yl)benzofuran - 2 - carboxylic acid (Compound 5)

[0096]

[0097] 1. Synthesis of Compound 5-2

[0098] Dissolve Compound 5-1 (2 g, 13.3 mmol, 1 eq), cupric acetate anhydrous (4.844 g, 26.7 mmol, 2 eq), and 4-bromopyrazole (1.96 g, 13.3 mmol, 1 eq) in 30 ml of dichloromethane. Add pyridine (2.15 ml, 26.7 mmol, 2 eq) and stir at room temperature for 4 h. Distill under reduced pressure, triturate, and column chromatograph to obtain Compound 5-2 (1.54 g, yield: 46.31%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.76 (d, J = 0.7 Hz, 1H), 7.89 - 7.83 (m, 1H), 7.79 - 7.69 (m, 2H), 7.40 - 7.31 (m, 2H), 2.66 (q, J = 7.6 Hz, 2H), 1.21 (t, J = 7.6 Hz, 3H).

[0099] 2. Synthesis of Compound 5-3

[0100] Add Compound 5-2 (0.4 g, 1.6 mmol, 1 eq), bis(pinacolato)diboron (0.406 g, 1.6 mmol, 1 eq), potassium acetate (0.314 g, 3.2 mmol, 2 eq), and Pd(dppf)Cl 2 (0.234 g, 0.32 mmol, 0.2 eq) to anhydrous 1,4-dioxane. Protect with N 2 and heat to 80 °C, stir for 16 h. Distill off the solvent under reduced pressure and proceed directly to the next step.

[0101] 3. The remaining steps are the same as in Example 1, except that Compound 5-3 is used instead of Compound 1-4.

[0102] Obtain Compound 5-4 (371 mg, yield: 86.9%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.14 (s, 1H), 8.33 (s, 1H), 8.04–7.93 (m, 2H), 7.81 (d, J = 8.3 Hz, 3H), 7.37 (d, J = 8.2 Hz, 2H), 3.91 (s, 3H), 2.66 (q, J = 7.6 Hz, 2H), 1.37 (s, 18H), 1.22 (t, J = 7.5 Hz, 3H).

[0103] Compound 5-5 was obtained (284.38 mg, yield: 97.89%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.90 (s, 1H), 8.29 (d, J = 1.8 Hz, 1H), 8.11 (s, 1H), 7.90 - 7.77 (m, 3H), 7.55 (d, J = 8.7 Hz, 1H), 7.39 (d, J = 8.3 Hz, 2H), 6.38 (s, 2H), 3.84 (s, 3H), 2.68 (q, J = 7.6 Hz, 2H), 1.24 (t, J = 2.7 Hz, 3H).

[0104] Compound 5-6 was obtained (396.49 mg, yield: 96.6%). 1 H NMR (300 MHz, CDCl 3 ) δ 8.56 (s, 1H), 8.50 (s, 1H), 8.42 (s, 1H), 7.93 - 7.87 (m, 2H), 7.75 (s, 1H), 7.72 - 7.66 (m, 3H), 7.53 - 7.47 (m, 2H), 7.17 - 7.11 (m, 2H), 3.88 (s, 2H), 2.67 (t, J = 1.0 Hz, 2H), 1.26 (s, 2H).

[0105] Compound 5 was obtained (368.08 mg, yield: 95.4%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.48 (s, 1H), 8.99 (s, 1H), 8.19 (d, J = 12.8 Hz, 2H), 8.05 (d, J = 8.2 Hz, 2H), 7.96 - 7.87 (m, 1H), 7.83 (d, J = 8.1 Hz, 2H), 7.76 (d, J = 8.7 Hz, 1H), 7.63 (d, J = 8.1 Hz, 2H), 7.36 (d, J = 8.1 Hz, 2H), 2.66 (q, J = 7.5 Hz, 2H), 1.36 (s, 9H), 1.22 (t, J = 7.5 Hz, 3H).

[0106] Example 6: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 6)

[0107]

[0108] The preparation method was the same as that of Example 5, except that Compound 6-1 was used instead of Compound 5-1.

[0109] Compound 6-2 (980 mg, yield: 53.4%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.98 (s, 1H), 8.08 (d, J = 8.5 Hz, 2H), 7.99 (s, 1H), 7.91 (d, J = 8.6 Hz, 2H).

[0110] Compound 6-3 was obtained and directly used for the next step.

[0111] Compound 6-4 (540 mg, yield: 96.8%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.36 (s, 1H), 8.48 (s, 1H), 8.15 (d, J = 8.4 Hz, 2H), 8.08 - 7.90 (m, 4H), 7.85 (d, J = 8.7 Hz, 1H), 3.92 (s, 3H), 1.38 (s, 18H).

[0112] Compound 6-5 (316.26 mg, yield: 87.73%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.12 (s, 1H), 8.32 (d, J = 1.8 Hz, 1H), 8.23 (s, 1H), 8.15 (d, J = 8.4 Hz, 2H), 7.94 (d, J = 8.5 Hz, 2H), 7.88 (dd, J = 8.7, 1.9 Hz, 1H), 7.57 (d, J = 8.7 Hz, 1H), 6.37 (s, 2H), 3.84 (s, 3H).

[0113] Compound 6-6 (387.62 mg, yield: 87.9%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.37 (s, 1H), 9.20 (s, 1H), 8.34 (s, 1H), 8.14 (t, J = 4.5 Hz, 3H), 8.04 (d, J = 8.1 Hz, 2H), 8.00 - 7.92 (m, 1H), 7.89 (d, J = 8.4 Hz, 2H), 7.79 (d, J = 8.7 Hz, 1H), 7.62 (d, J = 8.1 Hz, 2H), 3.88 (s, 3H), 1.35 (s, 9H).

[0114] Compound 6 (white solid, 317.96 mg, yield: 84.13%) was obtained. 1 H NMR (300 MHz, DMSO-d 6)δ 10.36 (s, 1H), 9.21 (s, 1H), 8.36 (s, 1H), 8.16 (d, J = 8.6 Hz, 3H), 8.05 (d, J = 8.1 Hz, 2H), 7.93 (t, J = 10.7 Hz, 3H), 7.79 (d, J = 8.6 Hz, 1H), 7.63 (d, J = 8.1 Hz, 2H), 1.37 (s, 9H).

[0115] Example 7: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 7)

[0116]

[0117] The preparation method was the same as that of Example 5, except that Compound 7-1 was used instead of Compound 5-1.

[0118] Compound 7-2 (650 mg, 64.13%) was obtained. 1 H NMR (300 MHz, CDCl 3 )δ 8.27 (s, 1H), 7.86 (s, 1H), 7.76 - 7.70 (m, 2H), 7.16 - 7.10 (m, 2H).

[0119] Compound 7-3 was obtained and directly used for the next step.

[0120] Compound 7-4 (310.45 mg, yield: 78.4%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 )δ 9.21 (s, 1H), 9.01 (s, 1H), 8.20 (s, 1H), 8.07 (d, J = 1.7 Hz, 1H), 7.98 - 7.80 (m, 2H), 7.71 (d, J = 8.7 Hz, 1H), 7.38 (t, J = 8.8 Hz, 2H), 3.85 (s, 3H), 1.48 (s, 9H).

[0121] Compound 7-5 (214.78 mg, yield: 88.9%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 )δ 8.92 (s, 1H), 8.27 (d, J = 1.9 Hz, 1H), 8.12 (s, 1H), 7.98 - 7.88 (m, 2H), 7.83 (dd, J = 8.7, 1.9 Hz, 1H), 7.54 (d, J = 8.7 Hz, 1H), 7.46 - 7.33 (m, 2H), 6.36 (s, 2H), 3.82 (s, 3H).

[0122] Compound 7-6 (265.8 mg, yield: 85%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.38 (s, 1H), 9.02 (s, 1H), 8.24 (s, 1H), 8.11 (s, 1H), 8.04 (d, J = 8.1 Hz, 2H), 7.97 - 7.90 (m, 3H), 7.78 (d, J = 8.8 Hz, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.37 (t, J = 8.7 Hz, 2H), 3.88 (s, 3H), 1.35 (s, 9H).

[0123] Compound 7 (217.93 mg, yield: 84.3%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.32 (s, 1H), 9.01 (s, 1H), 8.24 (s, 1H), 8.13 (d, J = 1.8 Hz, 1H), 8.04 (d, J = 8.2 Hz, 2H), 7.99 - 7.86 (m, 3H), 7.76 (d, J = 8.7 Hz, 1H), 7.62 (d, J = 8.3 Hz, 2H), 7.37 (t, J = 8.7 Hz, 2H), 1.35 (s, 9H).

[0124] Example 8: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(4-(trifluoromethoxy)phenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 8)

[0125]

[0126] The preparation method was the same as that of Example 5, except that Compound 8-1 was used instead of Compound 5-1.

[0127] Compound 8-2 (450 mg, 36.7%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.86 (s, 1H), 8.02 - 7.90 (m, 3H), 7.59 - 7.50 (m, 2H).

[0128] Compound 8-3 was obtained and directly used in the next step.

[0129] Compound 8-4 (210.7 mg, 89.4%) was obtained. 1 H NMR (300 MHz, CDCl 3) δ 8.56 (s, 1H), 8.50 (s, 1H), 8.42 (s, 1H), 7.93 - 7.87 (m, 2H), 7.77 - 7.71 (m, 3H), 7.69 (s, 1H), 7.53 - 7.47 (m, 2H), 7.29 - 7.23 (m, 2H), 3.88 (s, 2H).

[0130] Compound 8 - 5 (169.94 mg, yield: 94.5%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 8.99 (s, 1H), 8.29 (s, 1H), 8.16 (s, 1H), 8.03 (d, J = 8.5 Hz, 2H), 7.84 (d, J = 8.7 Hz, 1H), 7.56 (t, J = 7.5 Hz, 3H), 6.37 (s, 2H), 3.82 (s, 3H).

[0131] Compound 8 - 6 (203.89 mg, yield: 86.7%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 10.40 (s, 1H), 9.10 (s, 1H), 8.30 (s, 1H), 8.12 (d, J = 1.8 Hz, 1H), 8.04 (dd, J = 8.9, 3.1 Hz, 4H), 7.94 (dd, J = 8.7, 1.9 Hz, 1H), 7.79 (d, J = 8.7 Hz, 1H), 7.63 (d, J = 8.3 Hz, 2H), 7.54 (d, J = 8.6 Hz, 2H), 3.89 (s, 3H), 1.35 (s, 9H).

[0132] Compound 8 (white solid, 194.56 mg, yield: 97.8%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 10.33 (s, 1H), 9.08 (s, 1H), 8.28 (s, 1H), 8.14 (d, J = 1.8 Hz, 1H), 8.08–7.99 (m, 4H), 7.92 (dd, J = 8.7, 1.9 Hz, 1H), 7.77 (d, J = 8.7 Hz, 1H), 7.62 (d, J = 8.4 Hz, 2H), 7.54 (d, J = 8.6 Hz, 2H), 1.35 (s, 9H).

[0133] Example 9: Synthesis of 3-(4-(tert - butyl)benzamido)-5-(1 - phenyl - 1H - pyrazol - 4 - yl)benzofuran - 2 - carboxylic acid (Compound 9)

[0134]

[0135] The preparation method was the same as that of Example 5, except that compound 9-1 was used to replace compound 5-1.

[0136] Compound 9-2 (1.6 g, yield: 78.5%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.81 (s, 1H), 7.92 - 7.79 (m, 3H), 7.59 - 7.46 (m, 2H), 7.41 - 7.29 (m, 1H).

[0137] Compound 9-3 was obtained and directly used in the next step.

[0138] Compound 9-4 (349 mg, yield: 86%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.22 (s, 1H), 9.05 (s, 1H), 8.22 (s, 1H), 8.11 (d, J = 1.8 Hz, 1H), 7.97 - 7.87 (m, 2H), 7.72 (d, J = 8.7 Hz, 1H), 7.54 (t, J = 7.9 Hz, 1H), 7.40 - 7.29 (m, 1H), 3.88 (d, J = 4.0 Hz, 3H), 1.50 (s, 9H).

[0139] Compound 9-5 (240.99 mg, yield: 89.79%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.94 (s, 1H), 8.29 (d, J = 1.8 Hz, 1H), 8.13 (s, 1H), 7.93 - 7.88 (m, 2H), 7.85 (dd, J = 8.7, 1.8 Hz, 1H), 7.60 - 7.49 (m, 3H), 7.40 - 7.28 (m, 1H), 6.36 (s, 2H), 3.82 (s, 3H).

[0140] Compound 9-6 (272.96 mg, yield: 76.5%) was obtained. 1 H NMR (300 MHz, DMSO-d 6) δ 10.38 (s, 1H), 9.05 (s, 1H), 8.25 (s, 1H), 8.12 (d, J = 1.8 Hz, 1H), 8.09 - 8.00 (m, 2H), 8.00 - 7.86 (m, 3H), 7.78 (d, J = 8.7 Hz, 1H), 7.63 (d, J = 8.4 Hz, 2H), 7.52 (t, J = 7.8 Hz, 2H), 7.32 (t, J = 7.4 Hz, 1H), 3.88 (s, 3H), 1.35 (s, 9H).

[0141] Compound 9 (white solid, 256.72 mg, yield: 96.8%) was obtained. 1 1H NMR (300 MHz, DMSO-d 6 ) δ 10.32 (s, 1H), 9.03 (s, 1H), 8.24 (s, 1H), 8.14 (d, J = 1.8 Hz, 1H), 8.04 (d, J = 8.2 Hz, 2H), 7.97 - 7.87 (m, 3H), 7.76 (d, J = 8.7 Hz, 1H), 7.62 (d, J = 8.2 Hz, 2H), 7.52 (t, J = 7.8 Hz, 2H), 7.32 (t, J = 7.3 Hz, 1H), 1.35 (s, 9H).

[0142] Example 10: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(4-isopropylphenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 10)

[0143]

[0144] The preparation method was the same as that of Example 5, except that Compound 10-1 was used instead of Compound 5-1.

[0145] Compound 10-2 (2.4 g, yield: 73%) was obtained. 1 1H NMR (300 MHz, CDCl 3 ) δ 8.27 (s, 1H), 7.86 (s, 1H), 7.58 - 7.52 (m, 2H), 7.21 - 7.15 (m, 2H), 2.91 (t, J = 1.0 Hz, 1H), 1.26 (s, 6H).

[0146] Compound 10-3 was obtained and directly used for the next step.

[0147] Compound 10-4 (678.98 mg, yield: 89.79%) was obtained. 1 1H NMR (300 MHz, DMSO-d 6)δ8.91–8.84(m,1H),8.28(d,J=1.8Hz,1H),8.13-8.07(m,1H),7.89-.75(m,3H),7.53(d,J=8.7Hz,1H),7.40(d,J=8.5Hz,2H),6.36(s,2H),3.82(s,3H),2.96(p,J=6.9Hz,1H),1.56(s,9H),1.24(d,J=7.0Hz,6H).

[0148] Compound 10-5 (464.84 mg, yield: 86.72%) was obtained. 1 H NMR(300MHz,DMSO-d 6 )δ8.91–8.84(m,1H),8.28(d,J=1.8Hz,1H),8.13-8.07(m,1H),7.89-7.75(m,3H),7.53(d,J=8.7Hz,1H),7.40(d,J=8.5Hz,2H),6.36(s,2H),3.82(s,3H),2.96(p,J=6.9Hz,1H),1.24(d,J=7.0Hz,6H).

[0149] Compound 10-6 (234.1 mg, yield: 85%) was obtained. 1 H NMR(300MHz,DMSO-d 6 )δ10.42(s,1H),8.97(s,1H),8.17(d,J=16.2Hz,2H),8.03(d,J=8.0Hz,2H),7.91(d,J=9.1Hz,1H),7.82(d,J=8.3Hz,2H),7.75(d,J=8.8Hz,1H),7.62(d,J=8.1Hz,2H),7.38(d,J=8.2Hz,2H),3.89(s,3H),3.00-2.89(m,1H),1.35(s,9H),1.23(d,J=6.8Hz,6H).

[0150] Compound 10 (white solid, 141.6 mg, yield: 98.3%) was obtained. 1 H NMR(300MHz,DMSO-d 6)δ 10.42 (s, 1H), 8.97 (s, 1H), 8.17 (d, J = 16.2 Hz, 2H), 8.03 (d, J = 8.0 Hz, 2H), 7.91 (d, J = 9.1 Hz, 1H), 7.82 (d, J = 8.3 Hz, 2H), 7.75 (d, J = 8.8 Hz, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.38 (d, J = 8.2 Hz, 2H), 3.00 - 2.89 (m, 1H), 1.35 (s, 9H), 1.23 (d, J = 6.8 Hz, 6H).

[0151] Example 11: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(4-(tert-butyl)phenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 11)

[0152]

[0153] The preparation method was the same as that of Example 5, except that Compound 11-1 was used instead of Compound 5-1.

[0154] Compound 11-2 (1.13 g, yield: 67.9%) was obtained. 1 H NMR (300 MHz, CDCl 3 )δ 8.27 (s, 1H), 7.86 (s, 1H), 7.54 - 7.48 (m, 2H), 7.35 - 7.30 (m, 2H), 1.33 (s, 9H).

[0155] Compound 11-3 was obtained and directly used for the next step.

[0156] Compound 11-4 (342.14 mg, yield: 85%) was obtained. 1 H NMR (300 MHz, CDCl 3 )δ 8.58 (d, J = 1.8 Hz, 1H), 8.16 (s, 1H), 8.04 (d, J = 7.3 Hz, 1H), 7.66 (t, J = 8.2 Hz, 3H), 7.49 (dd, J = 8.7, 3.8 Hz, 4H), 4.02 (d, J = 2.0 Hz, 3H), 1.58 (s, 9H), 1.36 (s, 9H).

[0157] Compound 11-5 (259.1 mg, yield: 95.2%) was obtained. 1 H NMR (300 MHz, DMSO-d 6) δ 8.88 (s, 1H), 8.28 (d, J = 1.8 Hz, 1H), 8.10 (s, 1H), 7.89 - 7.75 (m, 3H), 7.60 - 7.49 (m, 3H), 6.36 (s, 2H), 3.82 (s, 3H), 1.33 (s, 9H).

[0158] Compound 11 - 6 (286.48 mg, yield: 78.34%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 11.76 (s, 1H), 8.92 (s, 1H), 8.47 (s, 1H), 8.15 (s, 1H), 8.00 (d, J = 8.2 Hz, 2H), 7.84 (d, J = 9.4 Hz, 3H), 7.61 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 8.3 Hz, 3H), 3.89 (s, 3H), 1.35 (s, 9H), 1.30 (s, 9H).

[0159] Compound 11 (white solid, 268.84 mg, yield: 96.3%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 ) δ 11.76 (s, 1H), 8.92 (s, 1H), 8.47 (s, 1H), 8.15 (s, 1H), 8.00 (d, J = 8.2 Hz, 2H), 7.84 (d, J = 9.4 Hz, 3H), 7.61 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 8.3 Hz, 3H), 1.34 (s, 9H), 1.32 (s, 9H).

[0160] Example 12: Synthesis of 3 - (4 - (tert - butyl)benzamido) - 5 - (1 - (4 - propylphenyl) - 1H - pyrazol - 4 - yl)benzofuran - 2 - carboxylic acid (Compound 12)

[0161]

[0162] The preparation method was the same as that of Example 5, except that Compound 12 - 1 was used instead of Compound 5 - 1.

[0163] Compound 12 - 2 (3.1 g, yield: 67.5%) was obtained. 1 H NMR (300 MHz, DMSO - d 6)δ 8.74 (s, 1H), 7.85 (s, 1H), 7.77 - 7.67 (m, 2H), 7.37 - 7.27 (m, 2H), 2.59 (t, J = 7.5 Hz, 2H), 1.70 - 1.51 (m, 2H), 0.90 (t, J = 7.3 Hz, 3H).

[0164] Compound 12 - 3 was obtained and directly used for the next step.

[0165] Compound 12 - 4 (906 mg, yield: 64.13%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 )δ 9.21 (s, 1H), 8.99 (s, 1H), 8.18 (s, 1H), 8.09 (d, J = 1.8 Hz, 1H), 7.89 (dd, J = 8.7, 1.9 Hz, 1H), 7.87 - 7.76 (m, 2H), 7.76 - 7.67 (m, 1H), 7.39 - 7.29 (m, 2H), 3.87 (s, 3H), 2.61 (t, J = 7.5 Hz, 2H), 1.63 (h, J = 7.4 Hz, 2H), 1.50 (s, 9H), 0.92 (t, J = 7.3 Hz, 3H).

[0166] Compound 12 - 5 (231.3 mg, yield: 83.1%) was obtained. 1 H NMR (300 MHz, DMSO - d 6 )δ 8.88 (s, 1H), 8.28 (d, J = 1.8 Hz, 1H), 8.10 (s, 1H), 7.83 - 7.75 (m, 3H), 7.53 (d, J = 8.7 Hz, 1H), 7.35 (d, J = 8.4 Hz, 2H), 6.36 (s, 2H), 3.82 (s, 3H), 2.61 (t, J = 7.6 Hz, 2H), 1.63 (h, J = 7.4 Hz, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0167] Compound 12 - 6 (249.15 mg, yield: 75.5%) was obtained. 1 H NMR (300 MHz, DMSO - d 6)δ 10.38 (s, 1H), 8.99 (s, 1H), 8.21 (s, 1H), 8.11 (d, J = 1.8 Hz, 1H), 8.09 - 8.00 (m, 2H), 7.98 - 7.73 (m, 4H), 7.68 - 7.58 (m, 2H), 7.32 (d, J = 8.4 Hz, 2H), 3.88 (s, 3H), 2.60 (t, J = 7.5 Hz, 2H), 1.62 (h, J = 7.0, 6.6 Hz, 2H), 1.35 (s, 9H), 0.91 (t, J = 7.3 Hz, 3H).

[0168] Compound 12 (white solid, 94.2 mg, yield: 96.8%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 )δ 10.42 (s, 1H), 8.97 (s, 1H), 8.17 (d, J = 12.9 Hz, 2H), 8.03 (d, J = 8.0 Hz, 2H), 7.90 (d, J = 9.1 Hz, 1H), 7.81 (d, J = 8.1 Hz, 2H), 7.75 (d, J = 8.6 Hz, 1H), 7.62 (d, J = 8.0 Hz, 2H), 7.32 (d, J = 8.1 Hz, 2H), 2.60 (t, J = 7.6 Hz, 2H), 1.66 - 1.55 (m, 2H), 1.35 (s, 9H), 0.91 (t, J = 7.3 Hz, 3H).

[0169] Example 13: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(3,4-difluorophenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 13)

[0170]

[0171] The preparation method was the same as that of Example 5, except that Compound 13-1 was used instead of Compound 5-1.

[0172] Compound 13-2 (421 mg, yield: 23.4%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 )δ 8.83 (s, 1H), 8.01 - 7.93 (m, 1H), 7.92 (s, 1H), 7.71 (dddd, J = 8.1, 4.2, 2.6, 1.4 Hz, 1H), 7.62 (dt, J = 10.3, 8.7 Hz, 1H).

[0173] Compound 13-3 was obtained and directly used in the next step.

[0174] Compound 13-4 (335 mg, yield: 85.49%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.23 (s, 1H), 9.08 (s, 1H), 8.26 (s, 1H), 8.09 (d, J = 1.8 Hz, 1H), 7.87 (td, J = 8.6, 1.9 Hz, 1H), 7.76 (dd, J = 15.2, 8.6 Hz, 3H), 7.70 - 7.58 (m, 1H), 3.87 (s, 3H), 1.50 (s, 9H).

[0175] Compound 13-5 (222.57 mg, yield: 94.3%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.98 (s, 1H), 8.32 - 8.24 (m, 1H), 8.15 (s, 1H), 8.02 (ddd, J = 12.0, 7.1, 2.6 Hz, 1H), 7.80 (ddd, J = 10.0, 8.2, 3.1 Hz, 2H), 7.72 - 7.58 (m, 1H), 7.55 (d, J = 8.6 Hz, 1H), 6.36 (s, 2H), 3.82 (d, J = 1.7 Hz, 3H).

[0176] Compound 13-6 (137.94 mg, yield: 96.21%) was obtained. 1 H NMR (300 MHz, CDCl 3 ) δ 8.56 (s, 1H), 8.54 (s, 1H), 8.50 (s, 1H), 7.93 - 7.87 (m, 2H), 7.77 - 7.72 (m, 2H), 7.69 (s, 1H), 7.53 - 7.47 (m, 2H), 7.38 (d, J = 2.0 Hz, 1H), 7.22 (d, J = 7.5 Hz, 1H), 3.88 (s, 3H), 1.35 (s, 9H).

[0177] Compound 13 (white solid, 95.4 mg, yield: 98%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.34 (s, 1H), 9.06 (s, 1H), 8.28 (s, 1H), 8.13 (s, 1H), 8.03 (d, J = 7.2 Hz, 4H), 7.79 (s, 2H), 7.62 (d, J = 8.3 Hz, 3H), 1.35 (s, 9H).

[0178] Example 14: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(4-cyanophenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 14)

[0179]

[0180] The preparation method was the same as that of Example 5, except that Compound 14-1 was used to replace Compound 5-1.

[0181] Compound 14-2 (775.94 mg, yield: 23.1%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.98 (s, 1H), 8.10 - 7.88 (m, 5H).

[0182] Compound 14-3 was obtained and directly used for the next step.

[0183] Compound 14-4 (312 mg, yield: 87.5%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.26 (d, J = 5.7 Hz, 2H), 8.36 (s, 1H), 8.18 - 8.07 (m, 3H), 8.04 (d, J = 8.8 Hz, 2H), 7.92 (dd, J = 8.8, 1.8 Hz, 1H), 7.76 (d, J = 8.7 Hz, 1H), 3.87 (s, 3H), 1.49 (s, 9H).

[0184] Compound 14-5 (208.69 mg, yield: 89%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.13 (s, 1H), 8.30 (s, 1H), 8.23 (s, 1H), 8.12 (d, J = 8.5 Hz, 2H), 8.04 (d, J = 8.4 Hz, 2H), 7.85 (d, J = 8.6 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 6.36 (s, 2H), 3.82 (s, 3H).

[0185] Compound 14-6 (194.83 mg, yield: 67.32%) was obtained. 11H NMR (300 MHz, DMSO-d6) δ 10.41 (s, 1H), 9.22 (s, 1H), 8.37 (s, 1H), 8.19 - 8.08 (m, 2H), 8.08 - 7.98 (m, 5H), 7.98 - 7.88 (m, 1H), 7.78 (dd, J = 8.8, 2.9 Hz, 1H), 7.62 (d, J = 8.3 Hz, 2H), 3.89 (s, 3H), 1.35 (s, 9H).

[0186] Compound 14 (white solid, 92.82 mg, yield: 95.4%) was obtained. 1 1H NMR (300 MHz, DMSO-d 6 ) δ 10.41 (s, 1H), 9.22 (s, 1H), 8.37 (s, 1H), 8.19 - 8.08 (m, 2H), 8.08 - 7.98 (m, 5H), 7.98 - 7.88 (m, 1H), 7.78 (dd, J = 8.8, 2.9 Hz, 1H), 7.62 (d, J = 8.3 Hz, 2H), 1.35 (s, 9H).

[0187] Example 15: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(p-tolyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 15)

[0188]

[0189] The preparation method was the same as that of Example 5, except that Compound 15-1 was used instead of Compound 5-1.

[0190] Compound 15-2 (936.56 mg, yield: 54%) was obtained. 1 1H NMR (300 MHz, DMSO-d 6 ) δ 8.75 (s, 1H), 7.85 (s, 1H), 7.76 - 7.65 (m, 2H), 7.31 (d, J = 8.2 Hz, 2H), 2.34 (s, 3H).

[0191] Compound 15-3 was obtained and directly used for the next step.

[0192] Compound 15-4 (314 mg, yield: 78.5%) was obtained. 1 1H NMR (300 MHz, CDCl 3) δ 8.58 (d, J = 2.0 Hz, 2H), 8.16 (s, 1H), 8.02 (s, 1H), 7.66 - 7.62 (m, 3H), 7.53 (dd, J = 15.9, 8.7 Hz, 1H), 7.29 (d, J = 8.4 Hz, 2H), 2.41 (s, 3H), 1.58 (s, 9H).

[0193] Compound 15-5 (192.6 mg, yield: 82.7%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.89 (s, 1H), 8.28 (d, J = 1.9 Hz, 1H), 8.10 (s, 1H), 7.89 - 7.79 (m, 2H), 7.77 (d, J = 2.0 Hz, 1H), 7.65 - 7.49 (m, 1H), 7.34 (d, J = 8.2 Hz, 2H), 6.37 (s, 2H), 3.82 (d, J = 1.9 Hz, 3H), 2.36 (s, 3H).

[0194] Compound 15-6 (83.72 mg, yield: 57.3%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.37 (s, 1H), 8.98 (s, 1H), 8.21 (s, 1H), 8.11 (s, 1H), 8.04 (d, J = 8.1 Hz, 2H), 7.94 (d, J = 9.1 Hz, 1H), 7.78 (dd, J = 8.4, 5.0 Hz, 3H), 7.62 (d, J = 8.1 Hz, 2H), 7.31 (d, J = 8.0 Hz, 2H), 3.88 (s, 3H), 2.35 (s, 3H), 1.35 (s, 9H).

[0195] Compound 15 (white solid, 77.5 mg, yield: 95.21%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.37 (s, 1H), 8.97 (s, 1H), 8.17 (d, J = 15.6 Hz, 2H), 8.03 (d, J = 8.1 Hz, 2H), 7.91 (d, J = 8.8 Hz, 1H), 7.80 (s, 1H), 7.77 (s, 3H), 7.62 (d, J = 7.9 Hz, 2H), 7.31 (d, J = 8.1 Hz, 2H), 2.35 (s, 3H), 1.35 (s, 9H).

[0196] Example 16: Synthesis of 3-(4-(tert-Butyl)benzamido)-5-(1-(4-methoxyphenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 16)

[0197]

[0198] The preparation method was the same as that of Example 5, except that Compound 16-1 was used instead of Compound 5-1.

[0199] Compound 16-2 (875.2 mg, yield: 53%) was obtained. 1 H NMR (300 MHz, CDCl 3 ) δ 7.86 (s, 1H), 7.65 (s, 1H), 7.54 (d, J = 8.7 Hz, 2H), 6.97 (d, J = 8.6 Hz, 2H), 3.85 (s, 3H).

[0200] Compound 16-3 was obtained and directly used for the next step.

[0201] Compound 16-4 (245.7 mg, yield: 80.2%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.21 (s, 1H), 8.92 (s, 1H), 8.15 (s, 1H), 8.08 (d, J = 1.8 Hz, 1H), 7.93 - 7.77 (m, 3H), 7.71 (d, J = 8.7 Hz, 1H), 7.14 - 7.05 (m, 2H), 3.87 (s, 3H), 3.82 (s, 3H), 1.50 (s, 11H).

[0202] Compound 16-5 (136.92 mg, yield: 87.32%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.81 (s, 1H), 8.26 (d, J = 1.8 Hz, 1H), 8.07 (s, 1H), 7.81 (dd, J = 9.4, 7.1 Hz, 3H), 7.52 (d, J = 8.7 Hz, 1H), 7.09 (d, J = 9.0 Hz, 2H), 6.34 (s, 2H), 3.82 (s, 4H), 3.82 (s, 3H).

[0203] Compound 16-6 (120.16 mg, yield: 83.4%) was obtained. 1 H NMR (300 MHz, DMSO-d 6)δ10.37(s,1H),8.92(s,1H),8.18(s,1H),8.10(d,J=1.8Hz,1H),8.04(d,J=8.1Hz,2H),7.92(dd,J=8.7,1.8Hz,1H),7.79(dd,J=12.2,8.8Hz,3H),7.62(d,J=8.1Hz,2H),7.07(d,J=9.0Hz,2H),3.88(s,3H),3.80(s,3H),1.35(s,9H).

[0204] Compound 16 (white solid, 93.21 mg, yield: 95.78%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 )δ10.83(s,1H),8.89(s,1H),8.25(s,1H),8.14(s,1H),8.02(d,J=8.2Hz,2H),7.84(td,J=8.9,2.0Hz,3H),7.71(d,J=8.7Hz,1H),7.61(d,J=8.1Hz,2H),7.12 - 7.03(m,2H),3.81(s,3H),1.35(s,9H).

[0205] Example 17: Synthesis of 3-(4-(tert-butyl)benzamido)-5-(1-(3,4-dimethylphenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 17)

[0206]

[0207] The preparation method was the same as that of Example 5, except that Compound 17-1 was used instead of Compound 5-1.

[0208] Compound 17-2 (901 mg, yield: 54.1%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 )δ8.71(s,1H),7.83(s,1H),7.63(d,J=2.4Hz,1H),7.52(dd,J=8.2,2.4Hz,1H),7.24(d,J=8.2Hz,1H),2.28(s,3H),2.24(s,3H).

[0209] Compound 17-3 was obtained and directly used for the next step.

[0210] Compound 17-4 (312 mg, yield: 87%) was obtained. 1 H NMR (300 MHz, DMSO-d 6)δ9.21(s,1H),8.96(s,1H),8.16(s,1H),8.09(s,1H),7.94 - 7.77(m,1H),7.75 - 7.49(m,3H),7.28(d,J=8.2Hz,1H),3.87(s,3H),2.32(s,3H),2.26(s,3H),1.50(s,9H).

[0211] Compound 17 - 5 (204.38 mg, yield: 87.2%) was obtained. 1 H NMR(300MHz,DMSO - d 6 )δ8.86(s,1H),8.28(d,J=1.8Hz,1H),8.09(s,1H),7.84(dd,J=8.7,1.8Hz,1H),7.70(d,J=2.4Hz,1H),7.63 - 7.51(m,2H),7.28(d,J=8.4Hz,1H),6.36(s,2H),3.82(s,3H),2.32(s,3H),2.26(s,3H).

[0212] Compound 17 - 6 (205.92 mg, yield: 71.34%) was obtained. 1 H NMR(300MHz,DMSO - d 6 )δ10.38(s,1H),8.97(s,1H),8.20(s,1H),8.10(d,J=1.8Hz,1H),8.04(d,J=8.4Hz,2H),7.94(dd,J=8.8,1.9Hz,1H),7.77(d,J=8.7Hz,1H),7.71(d,J=2.4Hz,1H),7.61(dd,J=9.3,7.3Hz,3H),7.26(d,J=8.2Hz,1H),3.88(s,3H),2.30(s,3H),2.25(s,3H),1.35(s,9H).

[0213] Compound 17 (white solid, 90.5 mg, yield: 93%) was obtained. 1 H NMR(300MHz,DMSO - d 6 )δ10.45(s,1H),8.95(s,1H),8.21–8.12(m,2H),8.03(d,J=8.3Hz,2H),7.90(dd,J=8.8,1.9Hz,1H),7.79 - 7.68(m,2H),7.62(d,J=8.1Hz,3H),7.25(d,J=8.2Hz,1H),2.30(s,4H),2.25(s,3H),1.35(s,9H).

[0214] Example 18: Synthesis of 5-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 18)

[0215]

[0216] 1. Synthesis of Compound 18-5

[0217] The synthesis method is the same as that of Compound 1-2, except that 5-bromo-2-hydroxybenzonitrile is replaced with 5-bromo-2-hydroxybenzaldehyde to obtain Compound 18-5 (1.27 g, yield: 67.89%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.07 - 8.00 (m, 1H), 7.73 (s, 2H), 7.67 (dd, J = 8.8, 2.0 Hz, 1H), 3.91 (s, 3H).

[0218] 2. Synthesis of Compound 18-6

[0219] The synthesis method is the same as that of Compound 2-5 to obtain Compound 18-6 (white solid, 268 mg, yield: 87.9%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.23 (s, 1H), 8.37 (s, 1H), 8.14 (dd, J = 5.3, 3.5 Hz, 3H), 7.93 (dd, J = 8.6, 2.1 Hz, 3H), 7.81 (d, J = 8.3 Hz, 2H), 3.92 (s, 3H).

[0220] 3. Synthesis of Compound 18

[0221] The synthesis method is the same as that of Compound 1 to obtain Compound 18 (187.83 mg, yield: 97.45%). 1 HNMR (300 MHz, DMSO-d 6 ) δ 9.23 (s, 1H), 8.37 (s, 1H), 8.19 - 8.10 (m, 3H), 7.91 (dd, J = 9.7, 7.9 Hz, 3H), 7.78 (d, J = 8.8 Hz, 1H), 7.67 (d, J = 0.9 Hz, 1H).

[0222] Example 19: Synthesis of 3-propionamide-5-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 19)

[0223]

[0224] The preparation method was the same as that of Example 6, except that 4-tert-butylbenzoyl chloride was replaced with propionyl chloride.

[0225] Compound 19-6 (197.16 mg, yield: 86.5%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.88 (s, 1H), 9.19 (s, 1H), 8.32 (d, J = 2.5 Hz, 1H), 8.16 (d, J = 8.5 Hz, 3H), 7.92 (d, J = 8.0 Hz, 4H), 7.74 (d, J = 8.8 Hz, 1H), 3.89 (s, 3H), 2.20 (s, 1H), 1.17 (t, J = 7.5 Hz, 3H).

[0226] Compound 19 (white solid, 95 mg, yield: 98%) was obtained. 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.88 (s, 1H), 9.19 (s, 1H), 8.32 (d, J = 2.5 Hz, 1H), 8.16 (d, J = 8.5 Hz, 3H), 7.92 (d, J = 8.0 Hz, 4H), 7.74 (d, J = 8.8 Hz, 1H), 2.20 (s, 1H), 1.17 (t, J = 7.5 Hz, 3H).

[0227] Example 20: Synthesis of 3-(cyclohexanecarboxamide)-5-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)benzofuran-2-carboxylic acid (Compound 20)

[0228]

[0229] The preparation method was the same as that of Example 6, except that 4-tert-butylbenzoyl chloride was replaced with cyclohexanecarbonyl chloride.

[0230] Compound 20-6 (203 mg, yield: 84%) was obtained. 1 H NMR (300 MHz, CDCl 3 ) δ 9.50 (s, 1H), 8.51 (d, J = 13.2 Hz, 2H), 8.42 (s, 1H), 7.84 - 7.77 (m, 2H), 7.75 (s, 1H), 7.69 (s, 1H), 7.67 - 7.61 (m, 2H), 3.88 (s, 3H), 2.52 (q, 1H), 1.95 (d, J = 13.0 Hz, 2H), 1.72 (d, J = 13.0 Hz, 2H), 1.59 - 1.50 (m, 3H), 1.38 (dd, J = 13.0, 8.2 Hz, 3H).

[0231] Compound 20 (white solid, 96.7 mg, yield: 96.5%) was obtained. 1 H NMR (300 MHz, CDCl 3 ) δ 9.53 (s, 1H), 8.51 (d, J = 13.2 Hz, 2H), 8.42 (s, 1H), 7.84 - 7.77 (m, 2H), 7.75 (s, 1H), 7.69 (s, 1H), 7.67 - 7.61 (m, 2H), 2.52 (q, 1H), 1.95 (d, J = 13.0 Hz, 2H), 1.72 (d, J = 13.0 Hz, 2H), 1.58 (d, J = 13.0 Hz, 1H), 1.53 (d, J = 13.0 Hz, 2H), 1.38 (dd, J = 13.0, 8.2 Hz, 3H).

[0232] Example 21 Biological Activity

[0233] The inhibitory effect of the compound on the cGAMP-activated STING pathway in THP1-Dual cells was detected by the luciferase method.

[0234] 1) 4×10 4 cells / well in the logarithmic growth phase were seeded in a 96-well plate, and different concentrations of the compound were added. The DMSO group was used as a blank control.

[0235] 2) The cells were transfected with a complex of 10 μg / mL cGAMP and lipofectamine 2000 (Invitrogen) and cultured for 24 h.

[0236] Preparation method of the transfection complex: 1 μg cGAMP was added to 10 μL Opit-MEM, 0.5 μL Lipofectamine2000 was added to 10 μL Opti-MEM, and after mixing, it was left standing for 15 minutes. Then 20 μL of the complex was added to the 96-well plate.

[0237] 3) The luciferase activity was detected using QUANTI-Luc TM reagent. 10 μL of the cell culture supernatant was taken and added to a 96-well opaque white plate, and then 50 μL of QUANTI-Luc TM reagent was added, and the reading was taken using a multifunctional microplate reader (Thermo).

[0238] Calculation method of relative luciferase activity: The cells treated with Lipofectamine-2000 were used as a blank control, and the cells treated with the Lipofectamine 2000:cGAMP complex were used as a negative control.

[0239] Relative luciferase activity = (RLU - RLUnon - transfected control) / (RLU negative control - RLUnon - transfected control), where RLU represents the raw luciferase value.

[0240] The IC of the compound was measured 50 as shown in Table 1 and compared with the positive control inhibitor H - 151:

[0241] Table 1 IC of the compound 50

[0242]

[0243] As can be seen from the above table, the above - mentioned compounds of the present invention and their pharmaceutically acceptable salts have good inhibitory effects on the cGAS - STING pathway, equivalent to or better than the positive control inhibitor H - 151, which can provide a basis for the preparation of drugs for the treatment and / or prevention of inflammatory diseases and autoimmune diseases.

[0244] Example 22 Evaluation of the binding ability of Compound 8 to STING by isothermal titration calorimetry (ITC)

[0245] The ITC experiment was performed using a MicroCal ITC 200 instrument to determine the binding affinity between a protein and a ligand. The ITC experiment can fit and quantitatively calculate the binding thermodynamic parameters, including the number of binding sites (n), the binding constant (Ka), the enthalpy change of the reaction (ΔH), the entropy change (ΔS), etc., by detecting the heat change (endothermic or exothermic) during the interaction between the protein solution and the small molecule (ligand) solution.

[0246] A high-concentration and high-purity STING-CTD protein was obtained through steps such as extraction, dialysis, and ultrafiltration. During dialysis, a buffer solution consistent with the protein component system was retained as much as possible. First, 400 μL of STING-CTD protein (1 μM) and 100 μL of the compound (10 μM) were prepared, and the DMSO content in the protein and the compound needed to be the same. Then, an isothermal titration calorimeter (MicroCal iTC200) was used, and the operating parameters were set: temperature 25 °C; rotation speed 750 rpm. The sample cell and the titration needle were cleaned. Approximately 200 μL of STING-CTD protein was slowly added to the sample cell, and approximately 50 μL of the compound was aspirated into the titration needle and dropped into the sample cell at a rate of 2 μL per drop, with a 2.5-minute interval between each drop, for a total of 19 drops. After the experiment, the 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 change ΔG of the binding of the compound to the STING-CTD protein, thereby obtaining the KA value and KD value (KD = 1 / KA). The smaller the KD value, the better the binding strength and selectivity of the compound to the protein.

[0247] The tested compound 8 showed a binding effect on STING, and the results were as Figure 1 shown. The binding affinity constant KD of compound 8 was 570 nM.

[0248] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as a limitation of the present invention itself. Various changes can be made in its form and details without departing from the spirit and scope of the present invention defined by the appended claims.

Claims

1. A 2-benzofuran carboxylic acid compound of general formula I or a pharmaceutically acceptable salt thereof: Wherein, A is selected from difluoromethyl, trifluoromethyl, ethyl or a substituted benzene ring; the substituted benzene ring is a benzene ring mono- or disubstituted by ortho- or para-methyl, ethyl, isopropyl, tert-butyl, halogen, n-propyl, methoxy, cyano, trifluoromethyl or trifluoromethoxy; B is selected from a benzene ring, a pyrazole ring or an imidazole ring; L C Selected from -NHCO; D is selected from ethyl, cyclohexyl or a benzene ring substituted with p-tert-butyl.

2. The compound according to claim 1, characterized in that selected from:

3. A method for preparing the compound according to any one of claims 1 to 2, characterized in that the synthetic route is as follows: wherein, A, B, L C , and D are as defined in General Formula I.

4. A pharmaceutical composition, characterized in that it comprises the compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

5. Use of the compound according to any one of claims 1 to 2 in the preparation of a drug for preventing and / or treating inflammatory diseases and autoimmune diseases.

6. The use according to claim 5, characterized in that the inflammatory diseases and autoimmune diseases are selected from systemic lupus erythematosus (SLE), familial chilblain lupus erythematosus (FCL), Aicardi-Goutières syndrome (AGS), STING-related infantile-onset vascular lesions (SAVI), Singleton-Merten syndrome (SMS), amyotrophic lateral sclerosis (ALS), Parkinson's syndrome (PD), non-alcoholic steatohepatitis (NASH), pulmonary fibrosis, aging, scleroderma, psoriasis, rheumatoid arthritis, inflammatory bowel disease, uveitis, mucositis, diabetes, cardiovascular diseases or neurodegenerative diseases.

7. The use according to claim 6, characterized in that the inflammatory bowel disease is autoimmune colitis, irritable bowel syndrome, ulcerative colitis, Crohn's disease.

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