Preparation method and application of sulfonamide compound for antagonizing NOD1

Abstract

The invention discloses a sulfonamide compound as well as a preparation method and application thereof. The compound can selectively antagonize human and mouse NOD1 signal channels, can inhibit secretion of inflammatory cytokines, and can be used for treating inflammatory diseases, especially gastritis caused by helicobacter pylori infection. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a novel sulfonamide organic small molecule compound, and a preparation method thereof. The compound can antagonize the activation of NOD1 signaling pathway and can be potentially used for the treatment of inflammatory diseases or gastric cancer and colorectal cancer, belonging to the field of medical technology. Background Art

[0002] Many diseases are closely related to the occurrence of inflammatory reactions and immune system disorders. Inflammation is an immune response of the body to damage caused by inflammatory factors, and is a defensive natural reaction. The human immune system is composed of innate immunity and adaptive immunity, which is the main functional system for the body to perform immune response, immune surveillance, and immune regulation. Innate immunity, also known as nonspecific immunity, is shared by most organisms including fungi, plants, insects, primitive multicellular organisms, and mammals. Mammals have evolved adaptive immunity, also known as specific immunity. Unlike the long-term effects of adaptive immunity on the host, innate immunity recognizes and responds to invading pathogens more quickly and is the basis of adaptive immune response. Innate immunity mainly recognizes pathogens through pattern recognition receptors, including Toll-like receptors (TLRs), C-type lectin receptors (CLRs), NOD-like receptors (NLRs), RIG-I-like receptors (RLRs) and DNA recognition receptors, which can recognize conservative pathogen-associated molecular patterns (PAMPs) that only exist in pathogens or danger-associated molecular patterns (DAMPs) released by endogenous tissue damage, such as lipopolysaccharide, peptidoglycan, nucleic acid, DNA, ATP, uric acid and other molecules. The expression locations of various pattern recognition receptors in cells are different. Most TLRs and CLRs are pattern recognition receptors expressed on the cell membrane, while a few TLRs, NLRs, RLRs and DNA recognition receptors are mainly expressed in the cytoplasm or nucleus.

[0003] NOD1 / 2 belongs to NLRs, and their proteins all have three common functional domains, including the leucine-rich repeat (LRR) at the carboxyl (C) terminal, which is functionally related to the recognition and binding of ligand pathogen-associated molecular patterns (PAMPs); the nucleotide binding domain (NACHT or NBD domain, also known as NOD domain) in the middle, which is related to the oligomerization activation of NOD1 / 2; and the caspase recruitment domain (CARD) at the amino (N) terminal, which is related to the recruitment of other proteins containing CARD domains. The N-terminus of NOD1 contains one CARD domain, while the N-terminus of NOD2 consists of two tandem CARD domains. NOD1 is widely expressed in various tissue cells, and NOD1 mainly recognizes iE-DAP (D-glutamyl-meso-diaminopimelic acid), the smallest structural unit of the cell wall of Gram-negative bacteria. After NOD1 is activated by iE-DAP, it will oligomerize and recruit the downstream node receptor-interacting protein 2 (RIP2) in an activated form. RIP2 is a serine / threonine protein kinase. After being recruited and activated, RIP2 continues to undergo phosphorylation, ubiquitination and other steps, and finally activates the NF-κB signaling pathway, MAPKs signaling pathway (including JNK, p38, ERK), and TRAF3 signaling pathway in the cell, triggering the release of downstream inflammatory factors and interferon. The NOD1 signaling pathway is involved in the occurrence and development of many microbial-related inflammatory diseases, such as rheumatoid arthritis, leprosy, atopic eczema, asthma, atopic dermatitis, sarcoidosis, Crohn's disease, tuberculosis, Blau syndrome, etc. Studies have shown that in addition to being associated with inflammatory and metabolic diseases, NOD1 also plays a role in the development of certain cancers. Chronic inflammation is often associated with the tumor microenvironment, also known as the tumor inflammatory microenvironment. This inflammatory microenvironment contains a large number of inflammatory factors, chemokines, and growth factors, and the immune inflammatory responses they produce can easily cause tumors to produce immunosuppression and immune escape, thereby promoting tumor development and metastasis.

[0004] Helicobacter pylori (hereinafter referred to as H.Pylori) is a Gram-negative bacterium. H.Pylori infection is the most common chronic infectious disease in the world, affecting approximately 4.4 billion people worldwide. NOD1 can recognize and bind to the peptidoglycan in the cell wall of H.Pylori, and after sending a signal through the caspase activation recruitment domain, it interacts with the CARD region of the downstream protein RIP2. NOD1 / RIP2 activates transcription factors NF-κB and MAPKs to produce pro-inflammatory cytokines and chemokines. Studies have found that H.Pylori can manipulate multiple cell receptors to interact with host cells, such as integrin-β2 (CD18), epidermal growth factor receptor (EGFR), CD74, TLRs, NLRP3, NOD1, NOD2, etc., but NOD1 is the most important mediating receptor. When patients are infected with H.Pylori, the NOD1-mediated NF-κB and MAPKs inflammatory signaling pathways are activated, producing a large number of inflammatory cytokines and chemokines. Patients will experience a development process of chronic gastritis-chronic atrophic gastritis-chronic atrophic gastritis with intestinal metaplasia (abbreviated as chronic gastritis with intestinal metaplasia)-dysplasia-gastric cancer. Therefore, it will be possible to treat H. Pylori-induced gastritis and prevent gastric cancer by selectively antagonizing the NOD1 signaling pathway.

[0005] Therefore, selectively targeting upstream targets or pathways of NF-kB signaling (such as NOD1) is expected to keep most innate immune defense mechanisms intact, but it is only effective for diseases where specific targets or pathways are causally related to the pathogenesis of the disease. However, most of the NOD1 small molecule antagonists reported so far have various shortcomings, including: 1. Poor selectivity. This causes other pattern recognition receptors to be antagonized, so the normal immune function of the human body is likely to be antagonized, which can lead to serious side effects or adverse reactions; 2. Low antagonistic activity. This results in the need for a larger dose, which may produce certain nonspecific toxicity; 3. Low oral bioavailability. 4. Poor metabolic stability. This results in a short drug retention time in the body and a high frequency of administration; 5. There are species differences. Some NOD1 antagonists only have an antagonistic effect on human NOD1 and are ineffective against mouse NOD1, which seriously limits their nonclinical pharmacodynamic studies. 6. No molecules can be used in clinical trials. Therefore, designing, synthesizing and optimizing NOD1 antagonists with good drugability, oral administration and high selectivity are expected to treat certain NOD1-related inflammatory diseases, especially chronic gastritis and even gastric cancer caused by H. Pylori. Summary of the invention

[0006] To solve the above problems, the present invention provides a sulfonamide compound, which has high activity, low toxicity, high selectivity for human NOD1 (hNOD1) and mouse NOD1 (mNOD1), can be orally administered to animals, and solves the species difference problem that may exist in such antagonists. Such compounds contain a sulfonamide compound as an active ingredient and at least one pharmaceutically acceptable carrier for preventing or treating inflammatory diseases (including gastric cancer), and a method for preparing the sulfonamide compound.

[0007] In order to achieve the above-mentioned object of the present invention, the present invention provides the following technical solutions:

[0008] A sulfonamide compound as shown in formula (I),

[0009]

[0010] In formula (I),

[0011] X is CH or N;

[0012] Y and Z are selected from hydrogen, methyl, methoxy, methylthio, trifluoromethylthio, dimethylamino, acetyl, ethylthio, amino, methylamino, ethylamino, morpholine, methanesulfonamide, halogen, nitro, trifluoromethyl, cyano, methanesulfonyl, nitrogen-containing heterocyclic ring, substituted aryl;

[0013] R1 is selected from vinyl, aryl, aryl derivatives, heterocycle, heterocycle derivatives, benzoheterocycle;

[0014] In some embodiments, the sulfonamide compound represented by formula (I) and its isomers, diastereomers, enantiomers and pharmaceutically acceptable salts are the following compounds:

[0015]

[0016] The compound of the present invention or the pharmaceutical composition containing it can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eyes, lungs and respiratory tract, skin, vaginal rectum, etc.

[0017] The compounds of the present invention were screened for biological activity against intracellular pattern recognition receptor NOD1 (including human NOD1 and mouse NOD1), and it was found that the sulfonamide compounds of the present invention can significantly antagonize the NOD1 signaling pathway. In addition, such compounds were also screened in other pattern recognition receptor (including NOD2, TLR4, RIP2) cell models, and it was found that the sulfonamide compounds of the present invention only have an antagonistic effect on NOD1, and therefore have high selectivity. Therefore, the compounds of the present invention can be used to prepare drug candidates for preventing or treating inflammation associated with NOD1 and related diseases.

[0018] The present invention discovers a novel NOD1 selective antagonist through rational drug design, skeleton transition, electronic isostere replacement and other medicinal chemistry strategies. The compound of the present invention can significantly antagonize the NOD1 signaling pathway and is useful for preparing drugs for preventing or treating inflammation and related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 , compound 17 antagonized the secretion of inflammatory cytokines induced by C12-iE-DAP;

[0020] Figure 2 , compound 17 antagonized the secretion of inflammatory cytokines induced by H. pylori;

[0021] Figure 3 , the structure of Compound A, a reported selective NOD1 antagonist, was used as a positive control drug;

[0022] Figure 4 , compound 17 inhibited the inflammatory response in mice induced by C12-iE-DAP;

[0023] Figure 5 , compound 17 inhibited H. Pylori-induced gastric inflammation in mice. DETAILED DESCRIPTION

[0024] In order to understand the present invention, the present invention is further illustrated by the following examples, but it is not intended to limit the protection scope of the present invention.

[0025] Embodiment 1:

[0026] Preparation of compound 1

[0027]

[0028] Under stirring at room temperature, 3-amino-4-chloroacetanilide (5 g, 27.2 mmol) was dissolved in 20 mL of pyridine, and then o-fluorobenzenesulfonyl chloride (6.3 g, 32.6 mmol) was slowly added and reacted at room temperature for 2 h. The reaction solution was washed three times with 1N hydrochloric acid solution, extracted with ethyl acetate, and the organic phases were combined and concentrated under reduced pressure to obtain 8.7 g of brown solid with a yield of 94%.

[0029]

[0030] The intermediate 1-a (5 g, 14.6 mmol) was dissolved in 20 mL of ethanol, and then 10 mL of concentrated hydrochloric acid was slowly added dropwise. The mixture was then heated to 80 °C and refluxed for 3 h. The mixture was concentrated under reduced pressure to remove most of the ethanol and concentrated hydrochloric acid. The pH was adjusted to neutral with sodium bicarbonate solution, and then extracted twice with ethyl acetate. The organic phases were combined and concentrated under reduced pressure to obtain 3.9 g of a yellow solid with a yield of 90%.

[0031]

[0032] Under nitrogen protection, intermediate 1-b (2 g, 6.7 mmol) was dissolved in 20 mL of ethyl acetate, and then triethylamine (2.3 mL, 16.7 mmol) was added. The reaction solution was then placed in an ice bath, and thiophosgene (616 μL, 8.1 mmol) was slowly added dropwise. The reaction was continued for 1 h, and the reaction solution was washed with saturated brine, extracted twice with ethyl acetate, and the organic phases were combined, concentrated, and column chromatography was performed to obtain 1.9 g of intermediate 1-c with a yield of 84%.

[0033]

[0034] The intermediate 1-c (100 mg, 0.29 mmol), 2-amino-5-chlorobenzoic acid (55 mg, 0.32 mmol) and triethylamine (101 μL, 0.73 mmol) were added to a reaction bottle containing 2 mL of 1,4-dioxane, and the reaction was carried out at 110° C. for 3 h. After the reaction, the mixture was washed with saturated brine, extracted with ethyl acetate, and the organic phase was concentrated under reduced pressure to dryness. Then 1 mL of dichloromethane was added, and the solid was fully dispersed by ultrasound. Then, the solid was collected to obtain 95 mg of a white solid, which was 1-d, with a yield of 66%.

[0035] Synthesis of target compound 1

[0036] At room temperature, intermediate 1-d (50 mg, 0.1 mmol) was added to 1.5 mL of 1,4-dioxane, and then thiophosgene (20 μL, 0.25 mmol) was slowly added dropwise, and then heated to 107 ° C for 1 h. After the reaction was completed, the reaction solution was washed with saturated brine, extracted twice with ethyl acetate, and the organic phases were combined, concentrated, and column chromatography was performed to obtain 24 mg of the target compound with a yield of 48%. 1 HNMR(400MHz,DMSO)δ10.61(s,1H),8.05(d,J=2.4Hz,1H),7.94(dd,J=8.7,2.5H z,1H),7.71(m,2H),7.69–7.58(m,3H),7.47–7.40(m,2H),7.32(t,J=7.6Hz,1H). 13 C NMR (101MHz, DMSO) δ160.4,159.6,157.1,145.1,144.1,136.5,136.0,135.9,135.3,133.7,131.7, 131.0,130.6,129.9,129.0,128.8,128.4,127.7,127.6,125.7,124.8,124.7,122.2,117.5,117.3.

[0037] Embodiment 2:

[0038]

[0039]

[0040] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-5-bromobenzoic acid to obtain intermediate 2-d.

[0041] Target compound 2 According to the synthesis method of target compound 1 in Example 1, intermediate 2-d was reacted with thiophosgene to obtain target compound 2 in a yield of 46%. 1 HNMR (400MHz, DMSO) δ10.57(s,1H),8.18(s,1H),8.05(d,J=8.2Hz,1H),7.84–7.53(m,5H),7.43(t,J=9.1Hz,2H),7.32(t,J=7.3Hz,1H). 13 C NMR (101MHz, DMSO) δ160.2,159.6,157.0,145.3,144.1,137.9,136.4,135.9,135.8,133.8,130 .9,130.5,129.9,128.8,128.8,128.3,127.7,127.6,124.7,124.6,122.4,119.8,117.4,117.2.

[0042] Embodiment 3:

[0043]

[0044] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-4,5-difluorobenzoic acid to obtain intermediate 3-d.

[0045] According to the synthesis method of target compound 1 in Example 1, intermediate 3-d was reacted with thiophosgene to obtain target compound 3 with a yield of 52%. 1 HNMR(400MHz,DMSO)δ10.60(s,1H),8.08(dd,J=9.9,8.7Hz,1H),7.86(dt,J =14.6,7.3Hz,1H),7.76–7.52(m,4H),7.48–7.38(m,2H),7.36–7.27(m,1H). 13C NMR(101MHz,DMSO)δ160.01,159.9,159.6,157.0,155.3,155.2,152.8,1 52.7,150.1,150.0,147.6,147.5,144.5,144.2,144.1,136.3,135.9,135 .8,133.7,130.9,130.5,129.9,128.9,128.3,127.7,127.6,124.7,124. 6,118.3,118.3,118.2,118.2,117.4,117.2,115.0,114.9,114.6,114.4.

[0046] Embodiment 4:

[0047]

[0048]

[0049] Under nitrogen protection, 2-amino-5-iodobenzoic acid (500 mg, 1.9 mmol), m-chlorophenylboronic acid (357 mg, 2.28 mmol), and sodium carbonate (504 mg, 4.75 mmol) were dissolved in a mixed solution of 4 mL 1,4-dioxane and 2 mL water, and the mixture was replaced with nitrogen. Tetrakis(triphenylphosphine)palladium (109 mg, 0.1 mmol) was added, and the nitrogen was replaced again. The mixture was heated to 100 ° C and reacted for 4 h. After the reaction was completed, the pH was adjusted to neutral, the mixture was extracted with ethyl acetate, the organic phase was concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 4-a with a yield of 77%.

[0050]

[0051] According to the synthesis method of 1-d in Example 1, intermediate 1-c reacts with intermediate 4-a to obtain intermediate 4-d.

[0052] Target compound 4 According to the synthesis method of target compound 1 in Example 1, intermediate 4-d was reacted with thiophosgene to obtain target compound 4 in a yield of 59%. 1 HNMR (400MHz, DMSO) δ10.58(s,1H),8.30(d,J=1.8Hz,1H),8.23(dd,J=8.5,1.9Hz,1H),7.82(s ,1H),7.79–7.66(m,4H),7.63(dd,J=8.6,5.4Hz,2H),7.57–7.40(m,4H),7.33(t,J=7.6Hz,1H). 13C NMR (101MHz, DMSO) δ161.1,159.6,157.0,145.9,143.6,140.5,137.6,136.6,135.9,135.8,133.9,133.7,133.7,130.9, 130.8,130.5,129.9,128.9,128.4,127.9,127.7,127.6,127.3,126.5,125.5,124.7,124.6,124.4,121.0,117.4,117.2.

[0053] Embodiment 5:

[0054]

[0055] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-5-nitrobenzoic acid to obtain intermediate 5-d.

[0056] Target compound 5 According to the synthesis method of target compound 1 in Example 1, intermediate 5-d was reacted with thiophosgene to obtain target compound 5 with a yield of 40%. 1 HNMR (400MHz, DMSO) δ10.61(s,1H),8.76(d,J=2.6Hz,1H),8.63(dd,J=8.9,2.7Hz,1H ),7.90(d,J=8.9Hz,1H),7.76–7.61(m,4H),7.48–7.39(m,2H),7.33(t,J=7.6Hz,1H). 13 C NMR (101MHz, DMSO) δ160.5,159.6,157.1,150.3,147.5,145.4,136.2,136.0,135.9,133.9,131.1, 130.7,129.9,129.2,128.8,128.4,128.2,127.7,127.6,124.7,124.7,122.5,121.3,117.4,117.2.

[0057] Embodiment 6:

[0058]

[0059] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-4,5-dichlorobenzoic acid to obtain intermediate 6-d.

[0060] According to the synthesis method of target compound 1 in Example 1, intermediate 6-d was reacted with thiophosgene to obtain target compound 6 in a yield of 66%. 1HNMR (400MHz, DMSO) δ10.60(s,1H),8.22(s,1H),8.03(d,J=8.1Hz,1H),7.69(dt,J=15 .2,7.6Hz,2H),7.60(dd,J=12.6,5.4Hz,2H),7.46–7.35(m,2H),7.31(t,J=7.6Hz,1H). 13 C NMR (101MHz, DMSO) δ159.8,159.6,157.1,145.6,145.4,137.8,136.2,135.8,135.8,134.2,134.2, 130.8,130.5,130.0,129.9,128.6,128.5,128.2,127.9,127.7,124.7,124.6,121.0,117.4,117.2.

[0061] Embodiment 7:

[0062]

[0063] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-5-trifluoromethylbenzoic acid to obtain intermediate 7-d.

[0064] According to the synthesis method of target compound 1 in Example 1, intermediate 7-d was reacted with thiophosgene to obtain target compound 7 in a yield of 54%. 1 HNMR (400MHz, DMSO) δ10.59(s,1H),8.34(d,J=1.3Hz,1H),8.22(dd,J=8.6,2.1Hz,1H),7.91( t,J=8.3Hz,1H),7.75–7.60(m,4H),7.44(dt,J=6.4,3.7Hz,2H),7.32(dt,J=13.2,2.9Hz,1H). 13 C NMR (101MHz, DMSO) δ160.7,159.6,157.1,148.8,146.2,136.3,136.0,135.9,133.8,131.2,131.2,131.0,130.6, 129.9,128.9,128.3,128.1,127.7,127.6,127.5,127.2,126.9,124.7,124.7,124.1,124.0,121.2,117.4,117.2.

[0065] Embodiment 8:

[0066]

[0067] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-4-nitrobenzoic acid to obtain intermediate 8-d.

[0068] According to the synthesis method of target compound 1 in Example 1, intermediate 8-d was reacted with thiophosgene to obtain target compound 8 with a yield of 43%. 1 HNMR (400MHz, DMSO) δ10.58(s,1H),8.41(d,J=1.8Hz,1H),8.37–8.26(m,2H),7.76–7.61(m,4H),7.48–7.39(m,2H),7.32(t,J=7.6Hz,1H). 13 C NMR (101MHz, DMSO) δ160.3,159.6,157.0,151.4,146.6,146.0,136.2,135.9,135.8,133.8,131.0, 130.6,129.9,128.9,128.8,128.2,127.7,127.6,125.3,124.7,124.6,121.4,121.1,117.4,117.2.

[0069] Embodiment 9:

[0070]

[0071] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-aminobenzoic acid to obtain intermediate 9-d.

[0072] According to the synthesis method of target compound 1 in Example 1, intermediate 9-d was reacted with thiophosgene to obtain target compound 9 with a yield of 70%. 1 H NMR (400MHz, DMSO) δ8.25(dd,J=7.9,1.1Hz,1H),7.91–7.80(m,2H),7.71(d,J=8.1Hz,1H),7.63(d,J=2.4 Hz,1H),7.60-7.53(m,2H),7.52–7.43(m,2H),7.21(dd,J=17.6,8.3Hz,2H),6.97(dd,J=8.5,2.4Hz,1H). 13C NMR (101MHz, DMSO) δ161.7,160.2,157.7,146.6,143.2,136.7,136.3,136.2,135.6,134.4,131 .2,130.7,128.1,127.7,127.2,126.4,126.3,125.6,124.7,124.6,121.3,120.5,117.5,117.3.

[0073] Embodiment 10:

[0074]

[0075]

[0076] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-5-cyanobenzoic acid to obtain intermediate 10-d.

[0077] According to the synthesis method of the target compound 1 in Example 1, the intermediate 10-d was reacted with thiophosgene to obtain the target compound 10 with a yield of 65%. 1 H NMR (400MHz, DMSO) δ10.59(s,1H),8.53(d,J=1.8Hz,1H),8.27(dd,J=8.5,1.9Hz,1H),7 .84(d,J=8.5Hz,1H),7.75–7.61(m,4H),7.46-7.41(m,2H),7.32(dd,J=9.2,6.0Hz,1H). 13 C NMR (101MHz, DMSO) δ160.2,159.6,157.1,148.9,146.8,137.5,136.3,136.0,135.9,133.8,132.0,131 .0,130.6,129.9,128.9,128.2,127.9,127.7,127.6,124.7,124.7,121.6,117.8,117.3,117.2,109.7.

[0078] Embodiment 11:

[0079]

[0080] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-5-methoxybenzoic acid to obtain intermediate 11-d.

[0081] Target compound 9 According to the synthesis method of target compound 1 in Example 1, intermediate 11-d was reacted with thiophosgene to obtain target compound 11 with a yield of 77%. 1 H NMR (400MHz, DMSO) δ10.58(s,1H),7.74–7.60(m,4H),7.56(d,J=2.3Hz,1H),7.52–7.39(m,4H),7.32(t,J=7.6Hz,1H),3.88(s,3H). 13 C NMR (101MHz, DMSO) δ161.0,159.6,158.3,157.1,140.8,140.6,136.7,136.0,135.9,133.7,130.8,130 .5,129.9,129.1,128.5,128.2,127.7,127.6,124.7,124.7,124.3,121.5,117.4,117.2,107.1,55.8.

[0082] Embodiment 12:

[0083]

[0084]

[0085] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-5-methylsulfonebenzoic acid to obtain intermediate 12-d.

[0086] According to the synthesis method of target compound 1 in Example 1, intermediate 12-d was reacted with thiophosgene to obtain target compound 12 with a yield of 63%. 1 H NMR (400MHz, DMSO) δ10.60(s,1H),8.56(d,J=2.0Hz,1H),8.36(dd,J=8.5,2.1Hz,1H),7.92 (d,J=8.5Hz,1H),7.74-7.63(m,4H),7.45-7.41(m,2H),7.33(t,J=7.6Hz,1H),3.34(s,3H). 13 C NMR (101MHz, DMSO) δ160.6,159.6,157.0,149.4,146.7,139.1,136.3,136.0,135.9,133.8,132.8,131 .0,130.6,129.9,128.8,128.2,128.0,127.7,127.6,126.6,124.7,124.7,121.1,117.4,117.2,43.4.

[0087] Embodiment 13:

[0088]

[0089] According to the synthesis method of 1-d in Example 1, intermediate 1-c is reacted with 2-amino-4-methoxy-5-chlorobenzoic acid to obtain intermediate 13-d.

[0090] According to the synthesis method of target compound 1 in Example 1, intermediate 13-d was reacted with thiophosgene to obtain target compound 13 with a yield of 75%. 1 HNMR(400MHz,DMSO)δ10.57(s,1H),8.03(s,1H),7.75–7.65(m,2H),7.62(d,J=8.5Hz,1H ),7.57(d,J=2.4Hz,1H),7.47–7.40(m,2H),7.37(s,1H),7.36–7.29(m,1H),4.03(s,3H). 13 C NMR(101MHz,DMSO)δ159.8,159.6,157.0,147.1,144.3,136.4,135.9,135.8,133.7,130.8,130.4, 129.9,129.0,128.4,127.7,127.6,127.2,124.7,124.6,121.7,117.4,117.2,114.3,108.8,57.1.

[0091] Embodiment 14:

[0092]

[0093] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2,5-difluorobenzenesulfonyl chloride to obtain intermediate 14-a.

[0094]

[0095] According to the synthesis method of 1-b in Example 1, the intermediate 14-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 14-b.

[0096]

[0097] According to the synthesis method of 1-c in Example 1, intermediate 14-b is reacted with thiophosgene to obtain intermediate 14-c.

[0098]

[0099] According to the synthesis method of 1-d in Example 1, intermediate 14-c is reacted with 2-amino-5-trifluoromethoxybenzoic acid to obtain intermediate 14-d.

[0100] Target compound 14 According to the synthesis method of target compound 1 in Example 1, intermediate 14-d was reacted with thiophosgene to obtain target compound 14 with a yield of 56%. 1 HNMR (400MHz, DMSO) δ10.89(s,1H),7.95(s,1H),7.90(dd,J=8.9,2.2Hz,1H),7.84(d,J=8.9Hz,1H),7.76–7 .70(m,1H),7.67(d,J=8.6Hz,1H),7.63(d,J=2.3Hz,1H),7.48(dd,J=8.5,2.4Hz,1H),7.25(t,J=9.1Hz,2H). 13 C NMR (101MHz, DMSO) δ160.5,160.1,160.0,157.5,157.4,146.5,145.1,144.3,136.5,136.0,135.8,135.7,133.4,131 .3,130.6,129.4,129.2,128.7,128.4,122.1,121.2,118.7,118.0,117.8,117.7,117.5,113.5,113.4,113.2,113.2.

[0101] Embodiment 15:

[0102]

[0103] According to the synthesis method of 1-d in Example 1, intermediate 14-c was reacted with 2-amino-5-trifluoromethylbenzoic acid to obtain intermediate 15-d.

[0104] According to the synthesis method of target compound 1 in Example 1, intermediate 15-d was reacted with thiophosgene to obtain target compound 15 in a yield of 35%. 1 H NMR (400MHz, DMSO) δ10.90(s,1H),8.33(d,J=0.9Hz,1H),8.21(dd,J=8.6,2.0Hz,1H),7.89(d,J =8.5Hz,1H),7.77–7.67(m,2H),7.65(d,J=2.4Hz,1H),7.51–7.46(m,1H),7.25(t,J=9.0Hz,2H). 13C NMR (101MHz, DMSO) δ160.6,160.1,160.0,157.5,157.4,148.7,146.2,136.4,136.0,135.9,135.8,133.4,131.3,131.2,131.2, 130.6,129.3,128.7,128.0,127.5,127.2,124.9,124.0,124.0,122.2,121.2,117.8,117.7,117.5,113.5,113.4,113.2,113.2.

[0105] Embodiment 16:

[0106]

[0107] According to the synthesis method of 1-d in Example 1, intermediate 14-c is reacted with 2-amino-5-cyanobenzoic acid to obtain intermediate 16-d.

[0108] Target compound 6 According to the synthesis method of target compound 1 in Example 1, intermediate 16-d was reacted with thiophosgene to obtain target compound 16 with a yield of 32%. 1 H NMR (400MHz, DMSO) δ10.92(s,1H),8.53(d,J=1.8Hz,1H),8.27(dd,J=8.5,1.9Hz,1H),7.8 4(d,J=8.5Hz,1H),7.78–7.62(m,3H),7.48(dd,J=8.5,2.4Hz,1H),7.26(t,J=9.1Hz,2H). 13 C NMR (101MHz, DMSO) δ160.1,160.0,160.0,157.5,157.4,148.9,146.7,137.5,136.4,135.9,133.4,132 .0,131.4,130.7,129.3,128.7,127.9,121.6,117.8,117.6,113.5,113.5,113.3,113.3,109.7,106.9.

[0109] Embodiment 17:

[0110]

[0111] According to the synthesis method of 1-d in Example 1, intermediate 14-c was reacted with 2-amino-5-methylsulfonebenzoic acid to obtain intermediate 17-d.

[0112] Target compound 17 According to the synthesis method of target compound 1 in Example 1, intermediate 17-d was reacted with thiophosgene to obtain target compound 17 in a yield of 32%. 1 H NMR (400MHz, DMSO) δ10.93(s,1H),8.56(d,J=2.1Hz,1H),8.36(dd,J=8.6,2.2Hz,1H),7.92(d,J =8.6Hz,1H),7.78–7.63(m,3H),7.48(dd,J=8.5,2.3Hz,1H),7.26(t,J=9.1Hz,2H),3.34(s,3H). 13 C NMR (101MHz, DMSO) δ160.6,160.0,160.0,157.5,157.4,149.4,146.7,139.1,136.4,136.0,135.9,135.8, 133.5,132.8,131.4,130.7,129.3,128.7,128.0,126.6,121.1,117.7,113.5,113.5,113.3,113.3,43.4.

[0113] Embodiment 18:

[0114]

[0115] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2-cyanobenzenesulfonyl chloride to obtain intermediate 18-a.

[0116]

[0117] According to the synthesis method of 1-b in Example 1, the intermediate 18-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 18-b.

[0118]

[0119] According to the synthesis method of 1-c in Example 1, intermediate 18-b is reacted with thiophosgene to obtain intermediate 18-c.

[0120]

[0121] According to the synthesis method of 1-d in Example 1, intermediate 18-c is reacted with 2-amino-5-methylsulfonebenzoic acid to obtain intermediate 18-d.

[0122] According to the synthesis method of target compound 1 in Example 1, intermediate 18-d was reacted with thiophosgene to obtain target compound 18 with a yield of 35%. 1H NMR (400MHz, DMSO) δ10.83(s,1H),8.56(d,J=2.1Hz,1H),8.37–8.34(m,1H),8.13–8.07(m,1H),7.92(d,J=8.5Hz ,1H),7.88–7.81(m,3H),7.66(d,J=8.5Hz,1H),7.61(d,J=2.3Hz,1H),7.47(dd,J=8.5,2.4Hz,1H),3.34(s,3H). 13 CNMR(101MHz,DMSO)δ160.6,149.3,146.7,141.0,139.1,136.4,136.0,133.6,133.5,13 3.4,132.8,131.6,130.7,129.5,129.2,128.7,128.0,126.5,121.1,115.6,109.3,43.4.

[0123] Embodiment 19:

[0124]

[0125] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 1,3-benzothiazole-6-sulfonyl chloride to obtain intermediate 19-a.

[0126]

[0127] According to the synthesis method of 1-b in Example 1, the intermediate 19-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 19-b.

[0128]

[0129] According to the synthesis method of 1-c in Example 1, intermediate 19-b is reacted with thiophosgene to obtain intermediate 19-c.

[0130]

[0131] According to the synthesis method of 1-d in Example 1, intermediate 19-c is reacted with 2-amino-5-methylsulfonebenzoic acid to obtain intermediate 19-d.

[0132] According to the synthesis method of target compound 1 in Example 1, intermediate 19-d was reacted with thiophosgene to obtain target compound 19 in a yield of 29%. 1HNMR (400MHz, DMSO) δ10.34(s,1H),9.62(s,1H),8.62(d,J=1.6Hz,1H),8.56(d,J=2.1Hz,1H),8.36(dd,J=8.6,2.1Hz,1H),8. 24(d,J=8.6Hz,1H),7.95–7.84(m,2H),7.62(d,J=8.5Hz,1H),7.57(d,J=2.3Hz,1H),7.41(dd,J=8.5,2.3Hz,1H),3.34(s,3H). 13 C NMR (101MHz, DMSO) δ161.0,160.7,155.3,149.4,146.7,139.1,136.3,136.3,134.3,134 .0,132.8,130.8,130.5,128.0,127.8,127.5,126.6,124.4,123.7,122.7,121.1,43.4.

[0133] Embodiment 20:

[0134]

[0135] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 4-methylformate benzenesulfonyl chloride to obtain intermediate 20-a.

[0136]

[0137] According to the synthesis method of 1-b in Example 1, the intermediate 20-a is deacetylated with thionyl chloride and methanol to obtain the intermediate 20-b.

[0138]

[0139] According to the synthesis method of 1-c in Example 1, intermediate 20-b is reacted with thiophosgene to obtain intermediate 20-c.

[0140]

[0141] According to the synthesis method of 1-d in Example 1, intermediate 20-c is reacted with 2-amino-5-methylsulfonebenzoic acid to obtain intermediate 20-d.

[0142] Target compound 20 According to the synthesis method of target compound 1 in Example 1, intermediate 20-d was reacted with thiophosgene to obtain target compound 20 with a yield of 37%. 1H NMR (400MHz, DMSO) δ10.44(s,1H),8.57(d,J=2.0Hz,1H),8.36(dd,J=8.6,2.1Hz,1H),8.11(d,J=8.4Hz,2H),7.92(d,J=8.6Hz, 1H),7.85(d,J=8.4Hz,2H),7.65(d,J=8.5Hz,1H),7.57(d,J=2.3Hz,1H),7.44(dd,J=8.5,2.3Hz,1H),3.88(s,3H),3.34(s,3H). 13 C NMR(101MHz,DMSO)δ165.0,160.6,149.3,146.7,143.5,139.0,136.3,133.8,13 3.2,132.7,130.7,130.7,129.9,128.1,127.9,127.1,126.4,121.0,52.5,43.3.

[0143] Embodiment 21:

[0144]

[0145] The intermediate 20-d (200 mg, 0.32 mmol) was dissolved in 2 mL of methanol, and then 1 mL of water was added, followed by NaOH (55 mg, 1.38 mmol). The mixture was reacted at room temperature for 6 h, and the pH was adjusted to acidic with dilute hydrochloric acid. The mixture was extracted three times with ethyl acetate, and the organic phases were combined and concentrated to obtain the intermediate 21-a as a white solid (165 mg) with a yield of 85%.

[0146]

[0147] The intermediate 21-a (150 mg, 0.27 mmol) was dissolved in 2 mL DMF, and then DIPEA (168 μL 1.06 mmol) was added, followed by HATU (154 mg, 0.41 mmol), and finally ethylenediamine biotin hydrochloride (105 mg, 0.32 mmol). The mixture was reacted at room temperature for h, and then washed with saturated brine three times, extracted with ethyl acetate, and the organic phases were combined, concentrated, and column chromatography was used to obtain the intermediate 21-b as a white solid (128 mg) with a yield of 57%. 1H NMR (800MHz, DMSO) δ13.40(s,1H),10.25(s,1H),8.70(s,1H),8.40(s,1H),8.27(d,J=8.3Hz,1H),7.98–7.89( m,3H),7.78(d,J=7.3Hz,2H),7.61(d,J=8.3Hz,1H),7.53(d,J=8.0Hz,1H),7.44(s,1H),7.21(d,J=7.9Hz,1H) ,6.42(s,1H),6.36(s,1H),4.30(s,1H),4.11(s,1H),3.30(s,5H),3.22(s,2H),3.06(s,1H),2.81(d,J=11.4H z,1H),2.58(d,J=12.4Hz,1H),2.07(d,J=7.6Hz,2H),1.60(s,1H),1.55–1.42(m,3H),1.30(d,J=18.5Hz,2H). 13 C NMR (201MHz, DMSO) δ176.7,172.3,165.0,162.7,158.9,142.7,141.8,138.1,135.8,133.9,133.5,130.3,129.1,128. 4,128.2,128.2,127.9,127.3,126.7,116.9,116.5,61.0,59.1,55.3,43.5,39.9,39.7,38.0,35.2,28.1,28.0,25.2.

[0148] Synthesis of target compound 21

[0149] According to the synthesis method of target compound 1 in Example 1, intermediate 21-b was reacted with thiophosgene to obtain target compound 21 with a yield of 27%. 1H NMR (800MHz, DMSO) δ10.38(s,1H),8.70(s,1H),8.58(s,1H),8.37(d,J=8.1Hz,1H),7.98(d,J=7.6Hz,2H), 7.96–7.90(m,2H),7.81(d,J=7.6Hz,2H),7.67(s,1H),7.64(d,J=8.3Hz,1H),7.44(d,J=8.1Hz,1H),6.42(s ,1H),6.36(s,1H),4.30(s,1H),4.11(s,1H),3.36(s,3H),3.31(s,2H),3.22(s,2H),3.06(s,1H),2.81(d,J =12.0Hz,1H),2.58(d,J=12.4Hz,1H),2.07(s,2H),1.60(s,1H),1.56–1.42(m,3H),1.30(d,J=18.1Hz,2H). 13 C NMR (201MHz, DMSO) δ172.8,165.5,163.1,161.2,149.9,147.3,142.2,139.6,138.8,136.9,134.6,133.3,131.3,130. 7,128.5,128.3,128.1,127.2,127.1,121.6,116.9,61.4,59.6,55.8,43.8,40.4,40.2,38.5,35.7,28.6,28.5,25.6.

[0150] Embodiment 22:

[0151]

[0152] Intermediate 22-b

[0153]

[0154] According to the synthesis method of 21-b in Example 21, intermediate 21-a is reacted with (3AS,4S,6AR)-N-[2-[2-(2-aminoethoxy)ethoxy]ethyl]hexahydro-2-oxo-1H-thieno[3,4-D]imidazole-4-pentanamide to obtain intermediate 22-b. 1H NMR (800MHz, DMSO) δ13.39(s,1H),10.22(s,1H),8.75(s,1H),8.40(s,1H),8.27(d,J=8.3Hz,1H),7.96(d,J=7.5Hz,2H),7 .82(s,1H),7.78(d,J=7.5Hz,2H),7.61(d,J=8.4Hz,1H),7.53(d,J=8.1Hz,1H),7.43(s,1H),7.22(d,J=8.1Hz,1H),6.39( d,J=43.9Hz,2H),4.31(s,1H),4.13(s,1H),3.53(d,J=18.9Hz,6H),3.41(d,J=24.3Hz,4H),3.30(s,3H),3.18(s,2H),3.0 9(s,1H),2.82(d,J=11.5Hz,1H),2.58(d,J=12.4Hz,1H),2.06(s,2H),1.61(s,1H),1.55–1.42(m,3H),1.35–1.25(m,2H). 13 C NMR (201MHz, DMSO) δ177.2,172.6,165.4,163.1,159.4,143.1,142.2,138.6,138.5,136.3,134.2,134.0,130.8,129.7,128.9 ,128.8,128.4,127.8,127.3,117.4,117.0,70.0,69.6,69.1,61.5,59.6,55.8,44.0,40.4,40.2,38.9,35.5,28.6,28.5,25.7.

[0155] Synthesis of target compound 22

[0156] According to the synthesis method of target compound 1 in Example 1, intermediate 22-b was reacted with thiophosgene to obtain target compound 22 with a yield of 23%. 1H NMR (800MHz, DMSO) δ10.37(s,1H),8.74(s,1H),8.57(s,1H),8.36(d,J=8.3Hz,1H),7.98(d,J=7.4Hz,2H),7.93(d,J =8.2Hz,1H),7.83–7.77(m,3H),7.64(d,J=14.1Hz,2H),7.43(d,J=8.2Hz,1H),6.38(d,J=44.1Hz,2H),4.30(s,1H), 4.12(s,1H),3.52(d,J=19.2Hz,6H),3.40(d,J=28.0Hz,4H),3.35(s,3H),3.17(d,J=3.1Hz,2H),3.08(s,1H),2.81( d,J=11.9Hz,1H),2.57(d,J=12.4Hz,1H),2.06(t,J=10.8Hz,2H),1.60(s,1H),1.55–1.41(m,3H),1.35–1.24(m,2H). 13 C NMR (201MHz, DMSO) δ172.1,164.9,162.7,160.7,149.4,146.8,141.8,139.1,138.2,136.4,134.1,132.8,130.8,130.3,128.0 ,127.9,127.7,126.8,126.6,121.1,118.0,69.5,69.1,68.7,61.0,59.1,55.4,43.4,39.9,39.7,38.4,35.0,28.1,28.0,25.2.

[0157] Embodiment 23:

[0158]

[0159] According to the synthesis method of 1-d in Example 1, intermediate 1-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 23-d.

[0160] According to the synthesis method of target compound 1 in Example 1, intermediate 23-d was reacted with thiophosgene to obtain target compound 23 with a yield of 36%. 1 H NMR (400MHz, DMSO) δ10.59(s,1H),8.95(s,1H),8.03(s,1H),7.76–7.58(m,4H),7.47–7.39(m,2H),7.32(t,J=7.6Hz,1H). 13C NMR (101MHz, DMSO) δ159.6,157.0,149.9,146.9,145.9,140.5,136.0,136.0,135.9,133.9, 131.0,130.7,129.9,129.3,128.6,128.0,127.6,127.5,124.7,124.7,119.2,117.4,117.2.

[0161] Embodiment 24:

[0162]

[0163]

[0164] According to the synthesis method of 1-d in Example 1, intermediate 1-c was reacted with 3-amino-6-chloropyridine-2-carboxylic acid to obtain intermediate 24-d.

[0165] According to the synthesis method of target compound 1 in Example 1, intermediate 24-d was reacted with thiophosgene to obtain target compound 24 with a yield of 32%. 1 H NMR (400MHz, DMSO) δ10.58(s,1H),8.20(d,J=8.6Hz,1H),7.98(d,J=8.6Hz,1H),7 .73-7.64(m,3H),7.60(d,J=2.3Hz,1H),7.47–7.38(m,2H),7.32(t,J=7.6Hz,1H). 13 C NMR (101MHz, DMSO) δ159.6,158.8,157.0,148.9,144.8,142.8,138.6,137.7,136.4,136.0,135 .9,133.8,130.9,130.7,130.4,129.9,128.5,128.1,127.7,127.5,124.7,124.7,117.4,117.2.

[0166] Embodiment 25:

[0167]

[0168] According to the synthesis method of 1-d in Example 1, intermediate 1-c was reacted with 2-amino-5-chloropyridine-3-carboxylic acid to obtain intermediate 25-d.

[0169] According to the synthesis method of target compound 1 in Example 1, intermediate 25-d was reacted with thiophosgene to obtain target compound 25 in a yield of 32%. 1H NMR (400MHz, DMSO) δ10.60 (s, 1H), 9.04 (d, J = 2.7Hz, 1H), 8.55 (d, J = 2.7Hz, 1H), 7.75–7.58 (m, 4H), 7.47–7.39 (m, 2H), 7.32 (dd, J = 11.3, 4.0Hz, 1H). 13 C NMR (101MHz, DMSO) δ160.9,159.6,157.1,154.8,154.6,147.7,136.2,136.0,135.9,135.0,133 .8,131.0,130.7,129.9,129.2,128.8,128.2,127.7,127.5,124.7,124.7,117.5,117.4,117.2.

[0170] Embodiment 26:

[0171]

[0172]

[0173] According to the synthesis method of 1-d in Example 1, intermediate 1-c was reacted with 3-aminoisonicotinic acid to obtain intermediate 26-d. Target compound 26 According to the synthesis method of target compound 1 in Example 1, intermediate 26-d was reacted with thiophosgene to obtain target compound 26 with a yield of 43%. 1 HNMR(400MHz,DMSO)δ10.57(s,1H),9.07(s,1H),8.75(d,J=5.1Hz,1H),7.98(d d,J=5.1,0.6Hz,1H),7.74–7.60(m,4H),7.47–7.39(m,2H),7.35–7.29(m,1H). 13 C NMR (101MHz, DMSO) δ160.5,159.6,157.0,149.4,146.9,145.5,141.0,136.3,135.9,135.9,133 .8,131.0,130.6,129.9,128.8,128.2,127.7,127.5,126.3,124.7,124.7,118.9,117.4,117.2.

[0174] Embodiment 27:

[0175]

[0176] According to the synthesis method of 1-d in Example 1, intermediate 1-c was reacted with 3-amino-6-chloropyrazine-2-carboxylic acid to obtain intermediate 27-d.

[0177] Target compound 26 was prepared by the synthesis method of target compound 1 in Example 1, and intermediate 27-d was reacted with thiophosgene to obtain target compound 27 in a yield of 26%. 1 H NMR (400MHz, DMSO) δ10.60(s,1H),9.18(s,1H),7.75–7.65(m,3H),7.60(d,J=2.4Hz,1H),7.48–7.37(m,2H),7.34(dd,J=11.2,4.0Hz,1H). 13 CNMR(101MHz,DMSO)δ159.5,159.4,157.0,151.5,150.5,148.2,146.2,136.1,136.0,135.9,134 .0,132.3,131.1,130.8,129.9,128.2,127.8,127.6,127.5,124.7,124.7,117.4,117.3,117.2.

[0178] Embodiment 28:

[0179]

[0180]

[0181] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with vinylsulfonyl chloride to obtain intermediate 28-a.

[0182]

[0183] According to the synthesis method of 1-b in Example 1, the intermediate 28-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 28-b.

[0184]

[0185] According to the synthesis method of 1-c in Example 1, intermediate 28-b is reacted with thiophosgene to obtain intermediate 28-c.

[0186]

[0187] According to the synthesis method of 1-d in Example 1, intermediate 28-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 28-d.

[0188] According to the synthesis method of target compound 1 in Example 1, intermediate 28-d was reacted with thiophosgene to obtain target compound 28 with a yield of 33%. 1 HNMR (400MHz, DMSO) δ9.92(s,1H),8.96(s,1H),8.06(d,J=6.2Hz,1H),7.74(d,J=8.5Hz,1H),7.64(d,J =2.3Hz, 1H), 7.41 (dd, J = 8.5, 2.3Hz, 1H), 6.84 (dd, J = 16.4, 9.9Hz, 1H), 6.07 (dd, J = 13.0, 11.5Hz, 2H). 13 C NMR (101MHz, DMSO) δ159.6,149.9,146.9,146.0,140.5,136.2,136.1,134.7,130.7,129.7,129.3,127.8,127.1,126.5,119.2.

[0189] Embodiment 29:

[0190]

[0191]

[0192] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 5-chlorothiophene-2-sulfonyl chloride to obtain intermediate 29-a.

[0193]

[0194] According to the synthesis method of 1-b in Example 1, the intermediate 29-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 29-b.

[0195]

[0196] According to the synthesis method of 1-c in Example 1, intermediate 29-b is reacted with thiophosgene to obtain intermediate 29-c.

[0197]

[0198] According to the synthesis method of 1-d in Example 1, intermediate 29-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 29-d.

[0199] According to the synthesis method of target compound 1 in Example 1, intermediate 29-d was reacted with thiophosgene to obtain target compound 29 in a yield of 39%. 1HNMR (400MHz, DMSO) δ10.63(s,1H),8.96(s,1H),8.05(s,1H),7.72(d,J=8.5Hz,1H),7.60( d,J=2.3Hz,1H),7.46(dd,J=8.5,2.4Hz,1H),7.36(d,J=4.1Hz,1H),7.23(d,J=4.1Hz,1H). 13 C NMR (101MHz, DMSO) δ159.6,149.9,146.9,146.0,140.5,138.2,136.2,135.7,133.8,132.6,131.2,131.0,129.3,128.3,128.1,119.2.

[0200] Embodiment 30:

[0201]

[0202]

[0203] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide is reacted with 2,3-dihydrobenzofuran-5-sulfonyl chloride to obtain intermediate 30-a.

[0204]

[0205] According to the synthesis method of 1-b in Example 1, the intermediate 30-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 30-b.

[0206]

[0207] According to the synthesis method of 1-c in Example 1, intermediate 30-b is reacted with thiophosgene to obtain intermediate 30-c.

[0208]

[0209] According to the synthesis method of 1-d in Example 1, intermediate 30-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 30-d.

[0210] According to the synthesis method of the target compound 1 in Example 1, the intermediate 30-d was reacted with thiophosgene to obtain the target compound 30 with a yield of 37%. 1H NMR (400MHz, DMSO) δ9.94(s,1H),8.96(s,1H),8.05(s,1H),7.63(d,J=8.5Hz,1H),7.59(d,J=2.1Hz,2H),7.50(dd, J=8.5,1.8Hz,1H),7.34(dd,J=8.5,2.4Hz,1H),6.86(d,J=8.5Hz,1H),4.62(t,J=8.9Hz,2H),3.19(t,J=8.8Hz,2H). 13 C NMR (101MHz, DMSO) δ163.4,159.7,149.9,146.9,146.0,140.6,136.1,134.9,130.9 ,130.7,129.6,129.4,128.6,128.3,127.0,126.3,124.4,119.2,109.0,72.2,28.3.

[0211] Embodiment 31:

[0212]

[0213]

[0214] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 1-methyl-1H-pyrazole-4-sulfonyl chloride to obtain intermediate 31-a.

[0215]

[0216] According to the synthesis method of 1-b in Example 1, the intermediate 31-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 31-b.

[0217]

[0218] According to the synthesis method of 1-c in Example 1, intermediate 31-b is reacted with thiophosgene to obtain intermediate 31-c.

[0219]

[0220] According to the synthesis method of 1-d in Example 1, intermediate 31-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 31-d.

[0221] According to the synthesis method of target compound 1 in Example 1, intermediate 31-d was reacted with thiophosgene to obtain target compound 31 in a yield of 25%. 1H NMR(400MHz,DMSO)δ9.96(s,1H),8.97(s,1H),8.22(s,1H),8.06(s,1H),7.73 –7.63(m,2H),7.59(d,J=2.3Hz,1H),7.36(dd,J=8.5,2.3Hz,1H),3.85(s,3H). 13 C NMR (101MHz, DMSO) δ159.7,149.9,146.9,146.1,140.6,137.99(s),136.1,135.0,133.2,130.9,129.4,129.1,126.7,125.3,120.7,119.2,39.0.

[0222] Embodiment 32:

[0223]

[0224]

[0225] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 3,5-dimethylisoxazole-4-sulfonyl chloride to obtain intermediate 32-a.

[0226]

[0227] According to the synthesis method of 1-b in Example 1, the intermediate 32-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 32-b.

[0228]

[0229] According to the synthesis method of 1-c in Example 1, intermediate 32-b is reacted with thiophosgene to obtain intermediate 32-c.

[0230]

[0231] According to the synthesis method of 1-d in Example 1, intermediate 32-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 32-d.

[0232] According to the synthesis method of target compound 1 in Example 1, intermediate 32-d was reacted with thiophosgene to obtain target compound 32 with a yield of 28%. 1 HNMR (400MHz, DMSO) δ10.61(s,1H),8.96(s,1H),8.05(s,1H),7.85–7.61(m,2H),7.51(d,J=7.1Hz,1H),2.30(d,J=37.3Hz,6H).13 C NMR (101MHz, DMSO) δ173.1,159.7,157.3,149.9,147.0,145.8,140.5,136.4,133.3,131.7,131.0,129.6,129.3,128.9,119.2,115.9,12.0,10.4.

[0233] Embodiment 33:

[0234]

[0235]

[0236] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2-thiophenesulfonyl chloride to obtain intermediate 33-a.

[0237]

[0238] According to the synthesis method of 1-b in Example 1, the intermediate 33-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 33-b.

[0239]

[0240] According to the synthesis method of 1-c in Example 1, intermediate 33-b is reacted with thiophosgene to obtain intermediate 33-c.

[0241]

[0242] According to the synthesis method of 1-d in Example 1, intermediate 33-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 33-d.

[0243] According to the synthesis method of target compound 1 in Example 1, intermediate 33-d was reacted with thiophosgene to obtain target compound 33 in a yield of 41%. 1 H NMR (400MHz, DMSO) δ10.42(s,1H),8.96(s,1H),8.05(s,1H),7.94(d,J=4.8Hz,1H),7.65(d,J =8.4Hz,1H),7.60(s,1H),7.47(d,J=3.6Hz,1H),7.37(d,J=8.2Hz,1H),7.13(t,J=3.9Hz,1H). 13C NMR (101MHz, DMSO) δ159.6,149.9,146.9,146.0,140.5,136.1,133.6,132.4,132.4,130.7,130.5,129.4,127.6,127.6,127.2,127.2,119.2.

[0244] Embodiment 34:

[0245]

[0246]

[0247] According to the synthesis method of 1-d in Example 1, intermediate 14-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 34-d.

[0248] According to the synthesis method of target compound 1 in Example 1, intermediate 34-d was reacted with thiophosgene to obtain target compound 34 with a yield of 40%. 1 H NMR(400MHz,DMSO)δ10.91(s,1H),8.95(s,1H),8.03(s,1H),7.78–7.66(m,2 H),7.62(d,J=2.3Hz,1H),7.45(dd,J=8.5,2.3Hz,1H),7.25(t,J=9.1Hz,2H). 13 C NMR (101MHz, DMSO) δ160.0,160.0,159.5,157.4,157.4,149.9,146.9,145.9,140.5,136.1, 135.9,133.6,131.3,130.7,129.3,129.0,128.5,119.2,117.6,113.5,113.4,113.2,113.2.

[0249] Embodiment 35:

[0250]

[0251] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2-chlorobenzenesulfonyl chloride to obtain intermediate 35-a.

[0252]

[0253] According to the synthesis method of 1-b in Example 1, the intermediate 35-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 35-b.

[0254]

[0255] According to the synthesis method of 1-c in Example 1, intermediate 35-b is reacted with thiophosgene to obtain intermediate 35-c.

[0256]

[0257] According to the synthesis method of 1-d in Example 1, intermediate 35-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 35-d.

[0258] According to the synthesis method of target compound 1 in Example 1, intermediate 35-d was reacted with thiophosgene to obtain target compound 35 in a yield of 46%. 1 H NMR (400MHz, DMSO) δ10.46 (s, 1H), 8.94 (s, 1H), 8.03 (s, 1H), 7.87 (d, J = 7.7Hz, 1H), 7.69–7. 60(m,3H),7.58(d,J=2.0Hz,1H),7.48(dd,J=10.2,4.2Hz,1H),7.38(dd,J=8.5,2.1Hz,1H). 13 C NMR(101MHz,DMSO)δ159.6,149.9,146.9,145.9,140.5,137.1,136.0,134 .5,134.1,131.9,131.0,130.9,130.7,129.3,127.9,127.8,127.5,119.2.

[0259] Embodiment 36:

[0260]

[0261] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with piperidinesulfonyl chloride to obtain intermediate 36-a.

[0262]

[0263] According to the synthesis method of 1-b in Example 1, the intermediate 36-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 36-b.

[0264]

[0265] According to the synthesis method of 1-c in Example 1, intermediate 36-b is reacted with thiophosgene to obtain intermediate 36-c.

[0266]

[0267] According to the synthesis method of 1-d in Example 1, intermediate 36-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 36-d.

[0268] According to the synthesis method of target compound 1 in Example 1, intermediate 36-d was reacted with thiophosgene to obtain target compound 36 in a yield of 24%. 1 HNMR(400MHz,DMSO)δ9.55(s,1H),8.95(d,J=0.6Hz,1H),8.04(d,J=0.7Hz,1H),7.72(d d,J=6.7,5.5Hz,2H),7.35(dd,J=8.5,2.4Hz,1H),3.23–3.02(m,4H),1.54–1.34(m,6H). 13 C NMR (101MHz, DMSO) δ159.6,149.9,146.9,146.0,140.5,136.1,135.8,130.6,129.4,128.1,126.2,125.5,119.2,46.5,24.8,23.0.

[0269] Embodiment 37:

[0270]

[0271] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2-trifluoromethylbenzenesulfonyl chloride to obtain intermediate 37-a.

[0272]

[0273] According to the synthesis method of 1-b in Example 1, the intermediate 37-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 37-b.

[0274]

[0275] According to the synthesis method of 1-c in Example 1, intermediate 37-b is reacted with thiophosgene to obtain intermediate 37-c.

[0276]

[0277] According to the synthesis method of 1-d in Example 1, intermediate 37-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 37-d.

[0278] The target compound 37 was prepared by reacting the intermediate 37-d with thiophosgene according to the synthesis method of the target compound 1 in Example 1, with a yield of 38%. 1H NMR (400MHz, DMSO) δ10.39(s,1H),8.94(s,1H),8.03(s,1H),7.99(d,J=7.5Hz,1H),7.91(d,J=7. 7Hz, 1H), 7.83 (dt, J=20.6, 7.2Hz, 2H), 7.63 (dd, J=13.3, 5.4Hz, 2H), 7.43 (dd, J=8.5, 2.3Hz, 1H). 13 C NMR (101MHz, DMSO) δ159.6,149.9,146.9,145.9,140.5,138.2,136.1,133.8,133.6,133.0, 131.3,131.3,130.7,129.3,128.8,128.4,128.3,128.2,126.5,126.2,125.9,125.5,119.2.

[0279] Embodiment 38:

[0280]

[0281] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 3-pyridinesulfonyl chloride to obtain intermediate 38-a.

[0282]

[0283] According to the synthesis method of 1-b in Example 1, the intermediate 38-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 38-b.

[0284]

[0285] According to the synthesis method of 1-c in Example 1, intermediate 38-b is reacted with thiophosgene to obtain intermediate 38-c.

[0286]

[0287] According to the synthesis method of 1-d in Example 1, intermediate 38-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 38-d.

[0288] According to the synthesis method of target compound 1 in Example 1, intermediate 38-d was reacted with thiophosgene to obtain target compound 38 with a yield of 26%. 1H NMR (400MHz, DMSO) δ10.54(s,1H),8.96(d,J=0.6Hz,1H),8.86–8.79(m,2H),8.10–8.03(m,2H),7.68–7.58(m,3H),7.41(dd,J=8.5,2.4Hz,1H). 13 CNMR(101MHz,DMSO)δ159.6,153.5,149.9,147.0,146.9,145.9,140.5,136 .2,136.1,134.6,134.0,130.9,130.5,129.3,127.9,127.8,124.2,119.2.

[0289] Embodiment 39:

[0290]

[0291] According to the synthesis method of 1-d in Example 1, intermediate 18-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 39-d.

[0292] According to the synthesis method of target compound 1 in Example 1, intermediate 39-d was reacted with thiophosgene to obtain target compound 39 in a yield of 38%. 1 H NMR (400MHz, DMSO) δ10.84(s,1H),8.95(d,J=0.5Hz,1H),8.13–8.08(m,1H),8.03(d,J=0.5Hz,1 H),7.87–7.83(m,3H),7.66(d,J=8.5Hz,1H),7.57(d,J=2.3Hz,1H),7.44(dd,J=8.5,2.4Hz,1H). 13 C NMR(101MHz,DMSO)δ159.6,149.9,146.9,146.0,141.0,140.5,136.2,136.0,133 .6,133.6,133.4,131.7,130.8,129.5,129.2,129.1,128.6,119.2,115.6,109.3.

[0293] Embodiment 40:

[0294]

[0295] According to the synthesis method of 1-d in Example 1, intermediate 19-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 40-d.

[0296] According to the synthesis method of the target compound 1 in Example 1, the intermediate 40-d was reacted with thiophosgene to obtain the target compound 40 with a yield of 30%. 1 HNMR (400MHz, DMSO) δ10.33(s,1H),9.62(s,1H),8.95(d,J=0.7Hz,1H),8.61(d,J=1.6Hz,1H),8.26–8.22(m,1H),8.02( d,J=0.7Hz,1H),7.87(dd,J=8.6,1.9Hz,1H),7.62(d,J=8.5Hz,1H),7.55(d,J=2.3Hz,1H),7.37(dd,J=8.5,2.4Hz,1H). 13 CNMR(101MHz,DMSO)δ161.0,159.6,155.3,149.9,146.9,145.9,140.5,136.2,136 .1,134.3,134.0,130.8,130.5,129.3,127.6,127.3,124.3,123.7,122.6,119.2.

[0297] Embodiment 41:

[0298]

[0299] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 6-chloropyridine-3-sulfonyl chloride to obtain intermediate 41-a.

[0300]

[0301] According to the synthesis method of 1-b in Example 1, the intermediate 41-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 41-b.

[0302]

[0303] According to the synthesis method of 1-c in Example 1, intermediate 41-b is reacted with thiophosgene to obtain intermediate 41-c.

[0304]

[0305] According to the synthesis method of 1-d in Example 1, intermediate 41-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 41-d.

[0306] According to the synthesis method of the target compound 1 in Example 1, the intermediate 41-d was reacted with thiophosgene to obtain the target compound 41 with a yield of 20%. 1H NMR (400MHz, DMSO) δ10.66(s,1H),8.97(d,J=0.6Hz,1H),8.68–8.66(m,1H),8.09(dd,J=8.4,2.6Hz,1H),8.05(d ,J=0.6Hz,1H),7.76(d,J=8.4Hz,1H),7.70(d,J=8.5Hz,1H),7.58(d,J=2.3Hz,1H),7.45(dd,J=8.5,2.4Hz,1H). 13 CNMR(101MHz,DMSO)δ159.6,154.2,149.9,147.7,146.9,145.9,140.5,138 .0,136.3,135.5,133.6,131.0,130.9,129.2,128.4,128.2,125.2,119.2.

[0307] Embodiment 42:

[0308]

[0309] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with (E)-2-phenylene-1-sulfonyl chloride to obtain intermediate 42-a.

[0310]

[0311] According to the synthesis method of 1-b in Example 1, the intermediate 42-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 42-b.

[0312]

[0313] According to the synthesis method of 1-c in Example 1, intermediate 42-b is reacted with thiophosgene to obtain intermediate 42-c.

[0314]

[0315] According to the synthesis method of 1-d in Example 1, intermediate 42-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 42-d.

[0316] According to the synthesis method of target compound 1 in Example 1, intermediate 42-d was reacted with thiophosgene to obtain target compound 42 in a yield of 38%. 1HNMR(400MHz,DMSO)δ9.92(s,1H),8.96(s,1H),8.01(s,1H),7.73–7.68(m,2H), 7.67–7.62(m,2H),7.45–7.38(m,4H),7.38–7.34(m,1H),7.25(d,J=15.4Hz,1H). 13 C NMR(101MHz,DMSO)δ159.6,149.9,146.9,146.0,141.5,140.5,136.1,134.8, 132.4,130.7,129.6,129.2,128.8,128.6,128.6,127.0,126.5,125.7,119.2.

[0317] Embodiment 43:

[0318]

[0319] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with N,N-dimethylsulfonyl chloride to obtain intermediate 43-a.

[0320]

[0321] According to the synthesis method of 1-b in Example 1, the intermediate 43-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 43-b.

[0322]

[0323] According to the synthesis method of 1-c in Example 1, intermediate 43-b is reacted with thiophosgene to obtain intermediate 43-c.

[0324]

[0325] According to the synthesis method of 1-d in Example 1, intermediate 43-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to obtain intermediate 43-d.

[0326] According to the synthesis method of target compound 1 in Example 1, intermediate 43-d was reacted with thiophosgene to obtain target compound 43 in a yield of 25%. 1 H NMR (400MHz, DMSO) δ9.61 (s, 1H), 8.96 (s, 1H), 8.05 (s, 1H), 7.75 (d, J = 2.3Hz, 1H), 7.72 (d, J = 8.5Hz, 1H), 7.38 (dd, J = 8.5, 2.4Hz, 1H), 2.73 (s, 6H). 13CNMR(101MHz,DMSO)δ159.7,149.9,146.9,146.1,140.6,136.2,135.8,130.7,129.4,129.0,126.7,126.5,119.2,37.8.

[0327] Embodiment 44:

[0328]

[0329] According to the synthesis method of 1-d in Example 1, intermediate 14-c was reacted with 3-aminoisonicotinic acid to obtain intermediate 44-d. Target compound 44 According to the synthesis method of target compound 1 in Example 1, intermediate 44-d was reacted with thiophosgene to obtain target compound 44 with a yield of 40%. 1 HNMR (400MHz, DMSO) δ10.92(s,1H),9.07(s,1H),8.74(d,J=5.1Hz,1H),7.97(d,J=5.1Hz,1H) ,7.78–7.66(m,2H),7.64(d,J=2.3Hz,1H),7.48(dd,J=8.5,2.4Hz,1H),7.26(t,J=9.1Hz,2H). 13 C NMR (101MHz, DMSO) δ160.6,160.0,160.0,157.5,157.4,149.4,147.0,145.5,141.1,136.4,136.0,135.9, 135.8,133.4,131.4,130.7,129.3,128.7,126.3,118.9,117.8,117.6,117.5,113.5,113.5,113.3,113.3.

[0330] Embodiment 45:

[0331]

[0332] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2,4-difluorobenzenesulfonyl chloride to obtain intermediate 45-a.

[0333]

[0334] According to the synthesis method of 1-b in Example 1, the intermediate 45-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 45-b.

[0335]

[0336] According to the synthesis method of 1-c in Example 1, intermediate 45-b is reacted with thiophosgene to obtain intermediate 45-c.

[0337]

[0338] According to the synthesis method of 1-d in Example 1, intermediate 45-c was reacted with 3-aminoisonicotinic acid to obtain intermediate 45-d. Target compound 45 According to the synthesis method of target compound 1 in Example 1, intermediate 45-d was reacted with thiophosgene to obtain target compound 45 with a yield of 42%. 1 H NMR(400MHz, CDCl3)δ9.13(s,1H),8.77(d,J=5.2Hz,1H),8.03–7.98(m,1H),7.83( td,J=8.7,6.1Hz,1H),7.68–7.61(m,2H),7.49(d,J=8.5Hz,1H),7.03–6.91(m,3H). 13 C NMR (101MHz, CDCl3) δ167.9,167.8,165.3,165.2,161.2,161.1,160.6,158.6,158.5,150.5,147.8,145.3,141.2,136.1,134.4,132 .9,132.8,130.9,126.5,125.7,125.7,123.0,122.9,122.8,122.8,121.8,119.3,112.3,112.3,112.1,112.1,106.4,106.1,105.9.

[0339] Embodiment 46:

[0340]

[0341] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2,5-difluorobenzenesulfonyl chloride to obtain intermediate 46-a.

[0342]

[0343] According to the synthesis method of 1-b in Example 1, the intermediate 46-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 46-b.

[0344]

[0345] According to the synthesis method of 1-c in Example 1, intermediate 46-b is reacted with thiophosgene to obtain intermediate 46-c.

[0346]

[0347] According to the synthesis method of 1-d in Example 1, intermediate 46-c was reacted with 3-aminoisocarboxylic acid to obtain intermediate 46-d. Target compound 46 According to the synthesis method of target compound 1 in Example 1, intermediate 46-d was reacted with thiophosgene to obtain target compound 46 in a yield of 39%. 1 HNMR (400MHz, DMSO) δ10.80(s,1H),9.08(s,1H),8.75(d,J=5.1Hz,1H),7.98(d,J=5.1Hz,1H ),7.68(d,J=8.5Hz,1H),7.65–7.58(m,2H),7.53(td,J=9.2,4.1Hz,1H),7.47-7.41(m,2H). 13 C NMR (101MHz, DMSO) δ160.6,158.2,155.8,153.3,149.4,147.0,145.5,141.1,136.4,133.5,131.3,129.0,128.9, 128.8,128.7,128.7,128.6,126.3,122.7,122.6,122.5,122.4,119.6,119.5,119.4,119.3,118.9,116.5,116.2.

[0348] Embodiment 47:

[0349]

[0350] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2,4,5-trifluorobenzenesulfonyl chloride to obtain intermediate 47-a.

[0351]

[0352] According to the synthesis method of 1-b in Example 1, the intermediate 47-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 47-b.

[0353]

[0354] According to the synthesis method of 1-c in Example 1, intermediate 47-b is reacted with thiophosgene to obtain intermediate 47-c.

[0355]

[0356] According to the synthesis method of 1-d in Example 1, intermediate 47-c was reacted with 3-aminoisonicotinic acid to obtain intermediate 47-d. Target compound 47 According to the synthesis method of target compound 1 in Example 1, intermediate 47-d was reacted with thiophosgene to obtain target compound 47 with a yield of 35%. 1 HNMR (400MHz, DMSO) δ10.83(s,1H),9.08(s,1H),8.75(d,J=5.1Hz,1H),7.97(d,J=5.1Hz,1H),7. 87(td,J=10.0,6.5Hz,1H),7.76–7.65(m,2H),7.55(d,J=2.1Hz,1H),7.48(dd,J=8.5,2.1Hz,1H). 13 C NMR (101MHz, DMSO) δ160.6,155.7,155.6,154.0,153.9,153.8,153.2,153.1,151.5,151.3,151.2,149.4,147.0,145.5,1 41.1,136.4,133.4,131.7,130.8,129.1,128.8,126.2,124.5,124.4,118.9,118.5,118.2,108.8,108.5,108.52,108.3.

[0357] Embodiment 48:

[0358]

[0359] According to the synthesis method of 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2-fluoro-4-nitrobenzenesulfonyl chloride to obtain intermediate 48-a.

[0360]

[0361] According to the synthesis method of 1-b in Example 1, the intermediate 48-a is deacetylated with concentrated hydrochloric acid to obtain the intermediate 48-b.

[0362]

[0363] According to the synthesis method of 1-c in Example 1, intermediate 48-b is reacted with thiophosgene to obtain intermediate 48-c.

[0364]

[0365] According to the synthesis method of 1-d in Example 1, intermediate 48-c is reacted with 3-aminoisonicotinic acid to obtain intermediate 48-d.

[0366] According to the synthesis method of target compound 1 in Example 1, intermediate 48-d was reacted with thiophosgene to obtain target compound 48 in a yield of 46%. 1 H NMR (400MHz, CDCl3) δ9.16 (s, 1H), 8.80 (d, J = 5.2Hz, 1H), 8.14–8.01 (m, 4H), 7.67 (d,J=2.4Hz,1H),7.56(s,1H),7.52(d,J=8.5Hz,1H),7.04(dd,J=8.5,2.4Hz,1H). 13 C NMR (101MHz, CDCl3) δ160.7,160.1,157.5,152.0,152.0,150.6,147.9,145.0,141.1,136.1,133 .7,132.4,132.3,132.3,131.1,127.0,126.4,125.7,122.5,119.6,119.6,119.3,113.5,113.2.

[0367] Embodiment 49:

[0368] Antagonistic effect of sulfonamide compounds on NOD1 in vitro

[0369] The NOD1 in vitro screening system used in the present invention is based on the HEK-Blue hNOD1 (human NOD1) cell line and the HEK-Blue mNOD1 (mouse NOD1) cell line in which cells highly express NOD1, and iE-DAP is used as an agonist of NOD1 to screen and obtain antagonists. In such screening cells, the SEAP reporter gene is integrated into the plasmid vector of the NF-κB promoter, so the activation or antagonism of the NOD1 signaling pathway by the compound can be detected by evaluating the secretion level of alkaline phosphatase.

[0370] Experimental methods:

[0371] The cells in the logarithmic growth phase were seeded in 96-well plates by adjusting the cell concentration to about 5 × 10 4 The compound to be screened was added to each well of cells, with the initial screening concentration of 1μM or 5μM, and only DMSO was added to the blank control group. After culturing in a 37°C, 5% CO2 incubator for 3 hours, NOD1 ligand C12-iE-DAP (final concentration of 50ng / mL) was added, and the culture was continued for 20 hours under the same conditions, and the OD value was detected at 655nm.

[0372] Inhibitory Percentage % = [(CT) / C] × 100, where C is the OD value of the blank control group, and T is the OD value of the compound group.

[0373] IC 50 Determination of IC value: The compound concentration has multiple gradients of 10μM, 1μM, 0.5μM, 0.1μM, 0.05μM, 0.01μM, 0.001μM and 0.0001μM. The operation method is the same as before. The antagonism rate (InhibitoryPercentage) of each concentration = [(CT) / C] × 100, fit the antagonist and concentration curve, and calculate IC 50 .

[0374] The activity screening results are shown in Table 1.

[0375] Table 1 Activity screening results of sulfonamide compounds

[0376]

[0377]

[0378] ND: Not Detected

[0379] The above screening results show that sulfonamide compounds have a certain inhibitory effect on the NOD1 signaling pathway of two species (human and mouse).

[0380] Embodiment 50

[0381] Four compounds were selected from Example 49 to study their selectivity, and the inhibitory effects of these four compounds on NOD2, and downstream node protein RIP2 and Toll-like receptor (hTLR4) were evaluated respectively. The experimental method was similar to that of Example 49. The results are shown in Table 2.

[0382] Table 2 Selectivity of sulfonamide compounds

[0383]

[0384]

[0385] From the above results, it can be seen that the preferred compounds of the present invention have very good selectivity and only have antagonistic effects on human and mouse NOD1.

[0386] Embodiment 51

[0387] Compound 17 antagonized the secretion of inflammatory cytokines induced by C12-iE-DAP.

[0388] After the NOD1 signaling pathway is activated, it can cause the secretion of inflammatory cytokines such as IL-6 and TNF-α. In this experiment, the transcription levels (mRNA) of IL-6 and TNF-α were measured by ELISA to evaluate the effect of compound 17 on the secretion of inflammatory cytokines mediated by the NOD1 signaling pathway. Figure 1 It can be seen that the mRNA levels of IL-6 and TNF-α in THP-1 and BMDM cells increased rapidly after C12-iE-DAP stimulation, while compound 17 could inhibit the mRNA levels of IL-6 and TNF-α to a state close to the initial state at 0.1 μM, indicating that compound 17 can antagonize the secretion of inflammatory cytokines mediated by NOD1 at a low dose.

[0389] Experimental methods:

[0390] (A) THP-1 cells were pretreated with compound 17 (0.1 μM or 1 μM) for 1 h, and the group without compound 17 was used as a control. Then, the NOD1-specific agonist C12-iE-DAP (1 μg / mL) was added and incubated for 2 h to activate the NOD1 signaling pathway to produce inflammatory factors, and finally, the mRNA levels of IL-6 and TNF-α were determined by q-RT-PCR. (B) BMDM cells were pretreated with compound 17 (0.1 μM or 1 μM) for 1 h, and the group without compound 17 was used as a control. Then, the NOD1-specific agonist C12-iE-DAP (20 μg / mL) was added and incubated for 2 h to activate the NOD1 signaling pathway to produce inflammatory factors, and finally, the mRNA levels of IL-6 and TNF-α were determined by q-RT-PCR. Data are expressed as mean ± SD (n = 3). Compared with cells stimulated with C12-iE-DAP alone, (*) p<0.05, (**) p<0.01, (***) p<0.001.

[0391] Embodiment 52

[0392] Compound 17 antagonized the secretion of inflammatory cytokines induced by H. Pylori.

[0393] H.Pylori colonizes in the stomach and activates the NOD1 signaling pathway of gastric cells, producing a large number of inflammatory factors, which eventually develop into gastritis or even gastric cancer. Therefore, we also evaluated whether compound 17 can antagonize the secretion of inflammatory cytokines induced by H.Pylori. The experimental results are as follows Figure 2 As shown, Figure 2A shows that after THP-1 cells were stimulated by C12-iE-DAP or H.Pylori, the mRNA levels of IL-6 and CXCL8 increased rapidly, and compound 17 at a concentration of 5 μM could inhibit the increase in IL-6 mRNA levels caused by H.Pylori stimulation to the initial level before stimulation, and also had a certain inhibitory effect on the expression level of CXCL8. Figure 2 B shows that in BMDM cells, compound 17 has an inhibitory effect of nearly 50% on the mRNA level of CXCL1 at a concentration of 5 μM, and also has a certain inhibitory effect on the mRNA level of TNF-α in a dose-dependent manner. This shows that compound 17 can also have a good antagonistic effect on the secretion of inflammatory cytokines induced by H. Pylori.

[0394] Experimental methods:

[0395] (A) THP-1 cells were pretreated with compound 17 (5 μM) for 1 h, and the group without compound 17 was used as a control. Then, the NOD1 specific agonist C12-iE-DAP (1 μg / mL) or H. Pylori (MOI=30) was added and incubated for 2 h to activate the NOD1 signaling pathway to produce inflammatory factors, and finally, the mRNA levels of IL-6 and CXCL8 were determined by q-RT-PCR. (B) BMDM cells were pretreated with compound 17 (1 μM or 5 μM) for 1 h, and the group without compound 17 was used as a control. Then, H. Pylori (MOI=30) was added and incubated for 2 h to activate the NOD1 signaling pathway to produce inflammatory factors, and finally, the mRNA levels of CXCL1 and TNF-α were determined by q-RT-PCR. Data are expressed as mean ± SD (n=3). Compared with cells stimulated with H. pylori alone, ns indicates no difference, (*) p<0.05, (**) p<0.01, (***) p<0.001.

[0396] Embodiment 53

[0397] Compound 17 inhibited the inflammatory response induced by C12-iE-DAP in mice.

[0398] In order to evaluate the anti-inflammatory effect of compound 17 in vivo, we established a mouse inflammation model in which NOD1 signaling pathway was activated by C12-iE-DAP stimulation. After intraperitoneal injection of C12-iE-DAP, the content of KC in the serum (a homolog of IL-8 in rodents) was significantly higher than that in the control group, indicating that the inflammation model was successfully established. The selective NOD1 antagonist Compound A reported in the literature was selected as the positive control drug. Its structure is shown in Figure 3 The experimental results are shown in Figure 4As shown, compound 17 inhibited the serum KC value induced by C12-iE-DAP stimulation in mice in a dose-dependent manner through oral administration, and the anti-inflammatory effect of compound 17 was equivalent to that of the positive compound Compound A at the same dose.

[0399] Experimental methods:

[0400] Compound 17 was pre-administered orally at doses of 12.5, 25 and 50 mg / kg, respectively. Compound A was used as a positive control. The negative control group and the blank control group were orally administered with the same volume of solvent. C12-iE-DAP (0.1 mg / kg) was intraperitoneally injected 15 minutes later, and the blank control group was intraperitoneally injected with an equal volume of solvent. Blood was collected 2 hours later, and the serum KC value of each group was determined by ELISF method. Data are expressed as mean ± SD (n = 5). Compared with the group given only C12-iE-DAP, ns indicates no difference, (*) p < 0.05, (**) p < 0.01, (***) p < 0.001.

[0401] Embodiment 54

[0402] Compound 17 inhibits H. Pylori-induced gastric inflammation in mice.

[0403] To further investigate whether compound 17 can also inhibit gastric inflammation in mice caused by H. Pylori, we first established an animal model of H. Pylori-infected mouse stomachs. 9 The bacterial solution with CFU / mL was gavaged at 0.2mL / 20g mouse body weight. After 3 days, the mouse stomach tissue was taken, homogenized and centrifuged, and the supernatant was taken to detect the KC content by Elisa. Compound 17 and the positive control Compound A were orally administered at five time points before and after modeling. The experimental results are shown in Figure 5 As shown, the KC value of gastric tissue increased significantly after H. Pylori infection (DMSO group), and the KC value of the positive control compound A (NOD1 antagonist reported in the literature) group was somewhat lower than that of the DMSO group. Compound 17 had a better inhibitory effect on gastric tissue KC at a dose of 12.5 mg / kg than the Compound A group. As the dosage of compound 17 increased, the inhibitory effect of KC became stronger, and the maximum inhibition rate was reached when 50 mg / kg was orally administered. The above results show that oral administration of compound 17 can inhibit gastric inflammation in mice caused by H. Pylori.

[0404] Experimental methods:

[0405] The mice in the drug-treated group were orally gavaged with compound 17 (12.5, 25, 50 mg / kg) and positive drug Compound A (25 mg / kg) 24 h and 2 h before modeling. At 0 h, the mice were gavaged with 0.2 mL of H. Pylori (HP) bacterial solution (1×10 9 CFU) / 20g to induce gastric inflammation. The same method was used for three times at 24h, 48h, and 72h after modeling. The mice were killed 2h after the last administration, and the gastric tissue was taken to determine the KC value. The negative control group (DMSO group) was modeled and given a blank solvent, while the blank control group (Ctrl group) was not modeled and only given a blank solvent. The data are expressed as mean ± SD (n = 4). Compared with the DMSO group, (*) p < 0.05, (**) p < 0.01, (***) p < 0.001.

Claims

1. A sulfonamide compound as shown in formula (I), In formula (I), X is CH or N; Y and Z are selected from hydrogen, methyl, methoxy, methylthio, trifluoromethylthio, dimethylamino, acetyl, ethylthio, amino, methylamino, ethylamino, morpholine, methanesulfonamide, halogen, nitro, trifluoromethyl, cyano, methanesulfonyl, nitrogen-containing heterocyclic ring, substituted aryl; R1 is selected from vinyl, aryl, aryl derivatives, heterocycle, heterocycle derivatives, and benzoheterocycle.

2. The sulfonamide compound and its isomers, diastereomers, enantiomers and pharmaceutically acceptable salts according to claim 1 are the following compounds:

3. A method for preparing the sulfonamide compound according to claim 1, comprising the following steps: Preparation method 1: The synthetic reagents and reaction conditions used are as follows: (i) sulfonyl chloride derivatives, pyridine, rt, 2h; (ii) concentrated hydrochloric acid, ethanol, 80℃, 3h or thionyl chloride, methanol, 70℃, 1h; (iii) thiophosgene, triethylamine, ethyl acetate, 0℃, 1h; (iv) anthranilic acid derivatives, triethylamine, 1,4-dioxane, 110℃, 3h or anthranilic acid ester derivatives, NaOH, DMSO, 130℃, 1h; (v) thiophosgene, 1,4-dioxane, 107℃, 1h.

4. Use of the sulfonamide compound according to any one of claims 1 to 2 in the preparation of a medicament for preventing or treating inflammatory diseases and related diseases.

5. According to the use of claim 4, the inflammatory-related diseases are gastric cancer and colorectal cancer.

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