Process for the preparation of sulfamide compounds antagonizing nod1 and uses thereof

Abstract

The application discloses a sulfonamide compound, a preparation method and application thereof. The compound can selectively antagonize a human and mouse NOD1 signal path, can inhibit secretion of inflammatory cytokines, and can be used for treating inflammatory diseases, in particular, gastritis caused by helicobacter pylori infection.

Description

TECHNICAL FIELD

[0001] The present application relates to a new type of sulfonamide small organic molecule compound, a preparation method thereof, and the compound can antagonize the activation of NOD1 signaling pathway, and can be potentially used for the treatment of inflammatory diseases or gastric cancer, colorectal cancer, and belongs to the technical field of medicine. BACKGROUND

[0002] Many diseases are closely related to the occurrence of inflammatory response and the disorder of immune system. Inflammation is an immune response of the body to the damage caused by inflammatory factors, and is a defensive natural reaction. The human immune system is composed of innate immunity and adaptive immunity, and is the main functional system of the body to perform immune response, immune surveillance and immune regulation. Innate immunity, also known as non-specific immunity, is common to 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 effect of adaptive immunity on the host, innate immunity is more rapid in recognizing and responding to invading pathogens, and is the basis of adaptive immune response. Innate immunity mainly plays its role in recognizing 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 NDA recognition receptors, which can recognize the conserved pathogen-associated molecular patterns (PAMPs) only present in pathogens or the danger-associated molecular patterns (DAMPs) released by endogenous tissue damage, such as lipopolysaccharide, peptidoglycan, nucleic acid, DNA, ATP, uric acid and other molecules. Different types of pattern recognition receptors are expressed in different locations in the cell. Most TLRs and CLRs are expressed in the cell membrane as pattern recognition receptors, while a few TLRs, NLRs, RLRs and DAN 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 related to the recognition and binding of ligand pathogen-associated molecular patterns (PAMP); 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 binding of other proteins containing the CARD domain. The N terminal of NOD1 contains one CARD domain, while the N terminal of NOD2 is composed of two tandem CARD domains. NOD1 is widely expressed in various tissues and cells, and mainly recognizes the minimum structural unit iE-DAP (D-glutamyl-meso-diaminopimelic acid) of the cell wall of gram-negative bacteria. After NOD1 is activated by iE-DAP, it will oligomerize to recruit downstream receptor-interacting protein 2 (RIP2). RIP2 belongs to serine / threonine protein kinase, and the activated RIP2 continues to undergo phosphorylation, ubiquitination and other steps, finally activates the NF-κB signaling pathway, MAPKs signaling pathway (including JNK, p38, ERK), and TRAF3 signaling pathway in the cell, and triggers the release of downstream inflammatory factors and interferons. 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, and Blau syndrome. Studies have shown that NOD1 is related to inflammatory diseases and metabolic diseases, and also plays a certain role in the development of some cancers. Chronic inflammation often accompanies the tumor microenvironment, also known as the tumor inflammatory microenvironment. There are a large number of inflammatory factors, chemokines and growth factors in this inflammatory microenvironment, and the immune inflammatory response they produce is easy to cause immune suppression and immune escape of tumors, thereby promoting the development and metastasis of tumors.

[0004] Helicobacter pylori (H. pylori) is a gram-negative bacterium, H. pylori infection is the most common chronic infectious disease in the world, affecting about 4.4 billion people worldwide. NOD1 can recognize and bind to peptidoglycan in the cell wall of H. pylori, interact with the CARD region of downstream protein RIP2 after signaling by caspase-activated recruitment domain, NOD1 / RIP2 activates transcription factors NF-κB and MAPKs, produces 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 mediator receptor. After the patient is infected with H. pylori, the NOD1-mediated NF-κB and MAPKs inflammatory signaling pathway is activated, and a large number of inflammatory cytokines and chemokines are produced. Patients will experience a development and evolution process from chronic gastritis-chronic atrophic gastritis-chronic atrophic gastritis with intestinal metaplasia (hereinafter referred to 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 only be effective for diseases in which a specific target or pathway has a causal relationship with the pathogenesis of the disease. However, most of the NOD1 small molecule antagonists reported so far have various shortcomings, including: 1. Poor selectivity. Other pattern recognition receptors are also antagonized, so the body's normal immune function is likely to be antagonized, which can cause serious side effects or adverse reactions; 2. Low antagonistic activity. It requires a large dose of medication, which can produce certain non-specific toxicity; 3. Low oral bioavailability; 4. Poor metabolic stability. It results in short drug retention time in the body and high dosing frequency; 5. Species difference. Some NOD1 antagonists only have antagonistic effect on human NOD1 and are ineffective on murine NOD1, which severely limits the non-clinical pharmacology research. 6. No molecules available for clinical trials. Therefore, designing and synthesizing NOD1 antagonists with good drug properties, oral availability, and high selectivity is expected to treat some inflammatory diseases related to NOD1, especially chronic gastritis and even gastric cancer caused by H. pylori. SUMMARY

[0006] To solve the aforementioned problems, the present application provides a sulfonamide compound, which has high activity, low toxicity, high selectivity for human NOD1 (hNOD1) and murine NOD1 (mNOD1), and can be orally administered, and solves the problem of species difference that may exist in such antagonists. The sulfonamide compound contains a sulfonamide compound as an active ingredient, and a pharmaceutical composition for preventing or treating an inflammatory disease (including gastric cancer) containing at least one pharmaceutically acceptable carrier, and a method for preparing the sulfonamide compound.

[0007] To achieve the above-mentioned object of the present application, the present application 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, methylsulfonyl, nitrogen-containing heterocycle, and substituted aryl;

[0013] R1 is selected from vinyl, aryl, aryl derivative, heterocycle, heterocycle derivative, and benzo-heterocycle;

[0014] In some embodiments, the sulfonamide compound shown in formula (I) and its isomers, diastereomers, enantiomers and pharmaceutical salts are the following compounds:

[0015]

[0016] The compound of the present application or the pharmaceutical composition containing it can be administered in unit dose form, and the administration route can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina rectum, etc.

[0017] The compound of the present application is screened for biological activity on intracellular pattern recognition receptor NOD1 (including human NOD1 and murine NOD1), and it is found that the sulfonamide compound of the present application can significantly antagonize the NOD1 signaling pathway. And this kind of compound is also screened in other pattern recognition receptor (including NOD2, TLR4, RIP2) cell models, and we find that the sulfonamide compound of the present application only has antagonistic effect on NOD1, and therefore has high selectivity. Therefore, the compound of the present application can be used for preparing a drug candidate for preventing or treating inflammation related to NOD1 and its related diseases.

[0018] The present application discloses a novel NOD1 selective antagonist by rational drug design, scaffold hopping, isosteric replacement and other medicinal chemistry strategies. The compounds of the present application can significantly antagonize NOD1 signaling pathway, and are useful for preparing drugs for preventing or treating inflammation and related diseases. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 Compound 17 antagonizes the secretion of inflammatory cytokines induced by C12-iE-DAP;

[0020] Figure 2 Compound 17 antagonizes the secretion of inflammatory cytokines induced by H. pylori;

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

[0022] Figure 4 Compound 17 inhibits the inflammatory response in mice induced by C12-iE-DAP;

[0023] Figure 5 Compound 17 inhibits the gastric inflammation in mice induced by H. pylori. DETAILED DESCRIPTION

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

[0025] Example 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. The reaction was carried out at room temperature for 2 h, and then the reaction solution was washed with 1N hydrochloric acid solution for 3 times. The organic phase was extracted with ethyl acetate, and then the organic phase was combined and concentrated under reduced pressure to obtain 8.7 g of brown solid with a yield of 94%.

[0029]

[0030] 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 reaction was carried out at 80℃ for 3 h, and then most of the ethanol and concentrated hydrochloric acid were removed by concentration under reduced pressure. The pH was adjusted to neutral with sodium bicarbonate solution, and then the solution was extracted with ethyl acetate for 2 times. The organic phase was combined and concentrated under reduced pressure to obtain 3.9 g of yellow solid with a yield of 90%.

[0031]

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

[0033]

[0034] 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, then the reaction was washed with saturated brine, and the organic phase was extracted with ethyl acetate and concentrated to dryness under reduced pressure, then 1 mL of dichloromethane was added, and the solid was dispersed by ultrasonic, then filtered, and the solid was collected to obtain 95 mg of white solid as 1-d, with a yield of 66%.

[0035] Synthesis of target compound 1

[0036] Intermediate 1-d (50 mg, 0.1 mmol) was added to 1.5 mL of 1,4-dioxane at room temperature, then sulfur trioxide (20 μL, 0.25 mmol) was slowly added dropwise, then heated to 107°C for 1 h, then the reaction was completed, and the reaction was washed with saturated brine, and the organic phase was extracted twice with ethyl acetate, and the organic phase was combined and concentrated to dryness, and column chromatography was performed to obtain 24 mg of target compound, with a yield of 48%. 1 HNMR (400 MHz, DMSO) δ 10.61 (s, 1H), 8.05 (d, J = 2.4 Hz, 1H), 7.94 (dd, J = 8.7, 2.5 Hz, 1H), 7.71 (m, 2H), 7.69 - 7.58 (m, 3H), 7.47 - 7.40 (m, 2H), 7.32 (t, J = 7.6 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.4, 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] Example 2:

[0038]

[0039]

[0040] Following the synthetic procedure of 1-d in Example 1, intermediate 1-c was reacted with 2-amino-5-bromobenzoic acid to give intermediate 2-d.

[0041] Target compound 2 was obtained following the synthetic procedure of target compound 1 in Example 1, reacting intermediate 2-d with thiophosgene to give target compound 2 in 46% yield. 1 HNMR (400 MHz, DMSO) δ 10.57 (s, 1H), 8.18 (s, 1H), 8.05 (d, J = 8.2 Hz, 1H), 7.84 - 7.53 (m, 5H), 7.43 (t, J = 9.1 Hz, 2H), 7.32 (t, J = 7.3 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.2, 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] Example 3:

[0043]

[0044] Following the synthetic procedure of 1-d in Example 1, intermediate 1-c was reacted with 2-amino-4,5-difluorobenzoic acid to give intermediate 3-d.

[0045] Target compound 3 was obtained following the synthetic procedure of target compound 1 in Example 1, reacting intermediate 3-d with thiophosgene to give target compound 3 in 52% yield. 1 HNMR (400 MHz, DMSO) δ 10.60 (s, 1H), 8.08 (dd, J = 9.9, 8.7 Hz, 1H), 7.86 (dt, J = 14.6, 7.3 Hz, 1H), 7.76 - 7.52 (m, 4H), 7.48 - 7.38 (m, 2H), 7.36 - 7.27 (m, 1H). 13C NMR (101 MHz, DMSO) δ 160.01, 159.9, 159.6, 157.0, 155.3, 155.2, 152.8, 152.7, 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] Example 4:

[0047]

[0048]

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

[0052] According to the synthesis method of compound 1 in Example 1, intermediate 4-d was reacted with thiophosgene to obtain target compound 4 with a yield of 59%. 1 HNMR (400MHz, DMSO) δ 10.58 (s, 1H), 8.30 (d, J = 1.8 Hz, 1H), 8.23 (dd, J = 8.5, 1.9 Hz, 1H), 7.82 (s, 1H), 7.79-7.66 (m, 4H), 7.63 (dd, J = 8.6, 5.4 Hz, 2H), 7.57-7.40 (m, 4H), 7.33 (t, J = 7.6 Hz, 1H). 13C NMR (101 MHz, DMSO) δ 161.1, 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] Example 5:

[0054]

[0055] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 2-amino-5-nitrobenzoic acid to give intermediate 5-d.

[0056] Target compound 5 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 5-d with thiophosgene to give target compound 5 in 40% yield. 1 HNMR (400 MHz, DMSO) δ 10.61 (s, 1H), 8.76 (d, J = 2.6 Hz, 1H), 8.63 (dd, J = 8.9, 2.7 Hz, 1H), 7.90 (d, J = 8.9 Hz, 1H), 7.76 - 7.61 (m, 4H), 7.48 - 7.39 (m, 2H), 7.33 (t, J = 7.6 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.5, 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] Example 6:

[0058]

[0059] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 2-amino-4,5-dichlorobenzoic acid to give intermediate 6-d.

[0060] Target compound 6 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 6-d with thiophosgene to give target compound 6 in 66% yield. 1HNMR (400 MHz, DMSO) δ 10.60 (s, 1H), 8.22 (s, 1H), 8.03 (d, J = 8.1 Hz, 1H), 7.69 (dt, J = 15.2, 7.6 Hz, 2H), 7.60 (dd, J = 12.6, 5.4 Hz, 2H), 7.46 - 7.35 (m, 2H), 7.31 (t, J = 7.6 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 159.8, 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] Example 7:

[0062]

[0063] Following the procedure for the synthesis of 1-d in Example 1, intermediate 1-c was reacted with 2-amino-5-trifluoromethylbenzoic acid to give intermediate 7-d.

[0064] Target compound 7 was obtained following the procedure for the synthesis of target compound 1 in Example 1, reacting intermediate 7-d with thiophosgene to give target compound 7 in 54% yield. 1 HNMR (400 MHz, DMSO) δ 10.59 (s, 1H), 8.34 (d, J = 1.3 Hz, 1H), 8.22 (dd, J = 8.6, 2.1 Hz, 1H), 7.91 (t, J = 8.3 Hz, 1H), 7.75 - 7.60 (m, 4H), 7.44 (dt, J = 6.4, 3.7 Hz, 2H), 7.32 (dt, J = 13.2, 2.9 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.7, 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] Example 8:

[0066]

[0067] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 2-amino-4-nitrobenzoic acid to give intermediate 8-d.

[0068] Target compound 8 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 8-d with thiophosgene to give target compound 8 in 43% yield. 1 HNMR (400 MHz, DMSO) δ 10.58 (s, 1H), 8.41 (d, J = 1.8 Hz, 1H), 8.37 - 8.26 (m, 2H), 7.76 - 7.61 (m, 4H), 7.48 - 7.39 (m, 2H), 7.32 (t, J = 7.6 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.3, 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] Example 9:

[0070]

[0071] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 2-amino-4-nitrobenzoic acid to give intermediate 8-d.

[0072] Target compound 9 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 9-d with thiophosgene to give target compound 9 in 70% yield. 1 H NMR (400 MHz, DMSO) δ 10.58 (s, 1H), 8.41 (d, J = 1.8 Hz, 1H), 8.37 - 8.26 (m, 2H), 7.76 - 7.61 (m, 4H), 7.48 - 7.39 (m, 2H), 7.32 (t, J = 7.6 Hz, 1H). 13C NMR (101 MHz, DMSO) δ 161.7, 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] Example 10:

[0074]

[0075]

[0076] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 2-amino-5-cyanobenzoic acid to give intermediate 10-d.

[0077] Target compound 10 was obtained following the synthetic procedure of Example 1 for compound 1, reacting intermediate 10-d with thiophosgene to give target compound 10 in 65% yield. 1 H NMR (400 MHz, DMSO) δ 10.59 (s, 1H), 8.53 (d, J = 1.8 Hz, 1H), 8.27 (dd, J = 8.5, 1.9 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.75 - 7.61 (m, 4H), 7.46 - 7.41 (m, 2H), 7.32 (dd, J = 9.2, 6.0 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.2, 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] Example 11:

[0079]

[0080] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 2-amino-5-methoxybenzoic acid to give intermediate 11-d.

[0081] Target compound 9 was obtained in 77% yield by reacting intermediate 11-d with thiophosgene according to the synthetic method of target compound 1 in Example 1. 1 H NMR (400 MHz, DMSO) δ 10.58 (s, 1H), 7.74 - 7.60 (m, 4H), 7.56 (d, J = 2.3 Hz, 1H), 7.52 - 7.39 (m, 4H), 7.32 (t, J = 7.6 Hz, 1H), 3.88 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 161.0, 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] Example 12:

[0083]

[0084]

[0085] Intermediate 12-d was obtained by reacting intermediate 1-c with 2-amino-5- methylsulfonylbenzoic acid according to the synthetic method of 1-d in Example 1.

[0086] Target compound 12 was obtained in 63% yield by reacting intermediate 12-d with thiophosgene according to the synthetic method of target compound 1 in Example 1. 1 H NMR (400 MHz, DMSO) δ 10.58 (s, 1H), 7.74 - 7.60 (m, 4H), 7.56 (d, J = 2.3 Hz, 1H), 7.52 - 7.39 (m, 4H), 7.32 (t, J = 7.6 Hz, 1H), 3.88 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 161.0, 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.

[0087] Example 13:

[0088]

[0089] Following the synthetic procedure of Example 1 for 1-d, intermediate 13-c was reacted with 2-amino-4-methoxy-5-chlorobenzoic acid to give intermediate 13-d.

[0090] Target compound 13 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 13-d with thiophosgene to give target compound 13 in 75% yield. 1 HNMR (400 MHz, DMSO) δ 10.57 (s, 1H), 8.03 (s, 1H), 7.75 - 7.65 (m, 2H), 7.62 (d, J = 8.5 Hz, 1H), 7.57 (d, J = 2.4 Hz, 1H), 7.47 - 7.40 (m, 2H), 7.37 (s, 1H), 7.36 - 7.29 (m, 1H), 4.03 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 159.8, 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] Example 14:

[0092]

[0093] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with 2,5-difluorobenzenesulfonyl chloride to give intermediate 14-a.

[0094]

[0095] Following the synthetic procedure of Example 1 for 1-b, intermediate 14-a was deacetylated with concentrated hydrochloric acid to give intermediate 14-b.

[0096]

[0097] Following the synthetic procedure of Example 1 for 1-c, intermediate 14-b was reacted with thiophosgene to give intermediate 14-c.

[0098]

[0099] Following the synthetic procedure of Example 1 for 1-d, intermediate 14-c was reacted with 2-amino-5-trifluoromethoxybenzoic acid to give intermediate 14-d.

[0100] Target compound 14 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 14-d with thiophosgene to give target compound 14 in 56% yield. 1 HNMR (400 MHz, DMSO) δ 10.89 (s, 1H), 7.95 (s, 1H), 7.90 (dd, J = 8.9, 2.2 Hz, 1H), 7.84 (d, J = 8.9 Hz, 1H), 7.76 - 7.70 (m, 1H), 7.67 (d, J = 8.6 Hz, 1H), 7.63 (d, J = 2.3 Hz, 1H), 7.48 (dd, J = 8.5, 2.4 Hz, 1H), 7.25 (t, J = 9.1 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 160.5, 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] Example 15:

[0102]

[0103] Following the synthetic procedure of Example 1 for 1-d, intermediate 14-c was reacted with 2-amino-5-trifluoromethylbenzoic acid to give intermediate 15-d.

[0104] Target compound 15 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 15-d with thiophosgene to give target compound 15 in 35% yield. 1 HNMR (400 MHz, DMSO) δ 10.90 (s, 1H), 8.33 (d, J = 0.9 Hz, 1H), 8.21 (dd, J = 8.6, 2.0 Hz, 1H), 7.89 (d, J = 8.5 Hz, 1H), 7.77 - 7.67 (m, 2H), 7.65 (d, J = 2.4 Hz, 1H), 7.51 - 7.46 (m, 1H), 7.25 (t, J = 9.0 Hz, 2H). 13C NMR (101 MHz, DMSO) δ 160.6, 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] Example 16:

[0106]

[0107] Following the synthetic procedure of Example 1 for 1-d, intermediate 14-c was reacted with 2-amino-5-cyanobenzoic acid to give intermediate 16-d.

[0108] Following the synthetic procedure of Example 1 for Compound 1, intermediate 16-d was reacted with thiophosgene to give Compound 16 in 32% yield. 1 H NMR (400 MHz, DMSO) δ 10.92 (s, 1H), 8.53 (d, J = 1.8 Hz, 1H), 8.27 (dd, J = 8.5, 1.9 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.78 - 7.62 (m, 3H), 7.48 (dd, J = 8.5, 2.4 Hz, 1H), 7.26 (t, J = 9.1 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 160.6, 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.

[0109] Example 17:

[0110]

[0111] Following the synthetic procedure of Example 1 for 1-d, intermediate 14-c was reacted with 2-amino-5-cyanobenzoic acid to give intermediate 16-d.

[0112] Target compound 17 was obtained by reacting intermediate 17-d with thiophosgene according to the synthetic method of target compound 1 in example 1 in 32% yield. 1 H NMR (400 MHz, DMSO) δ 10.93 (s, 1H), 8.56 (d, J = 2.1 Hz, 1H), 8.36 (dd, J = 8.6, 2.2 Hz, 1H), 7.92 (d, J = 8.6 Hz, 1H), 7.78 - 7.63 (m, 3H), 7.48 (dd, J = 8.5, 2.3 Hz, 1H), 7.26 (t, J = 9.1 Hz, 2H), 3.34 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 160.6, 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] Example 18:

[0114]

[0115] Intermediate 18-a was obtained by reacting 3-amino-4-chloroacetanilide with 2-cyanobenzenesulfonyl chloride according to the synthetic method of 1-a in example 1.

[0116]

[0117] Intermediate 18-b was obtained by removing acetyl group of intermediate 18-a with concentrated hydrochloric acid according to the synthetic method of 1-b in example 1.

[0118]

[0119] Intermediate 18-c was obtained by reacting intermediate 18-b with thiophosgene according to the synthetic method of 1-c in example 1.

[0120]

[0121] Intermediate 18-d was obtained by reacting intermediate 18-c with 2-amino-5-methylsulfonylbenzoic acid according to the synthetic method of 1-d in example 1.

[0122] Target compound 18 was obtained by reacting intermediate 18-d with thiophosgene according to the synthetic method of target compound 1 in example 1 in 35% yield. 1H NMR (400 MHz, DMSO) δ 10.83 (s, 1H), 8.56 (d, J = 2.1 Hz, 1H), 8.37 - 8.34 (m, 1H), 8.13 - 8.07 (m, 1H), 7.92 (d, J = 8.5 Hz, 1H), 7.88 - 7.81 (m, 3H), 7.66 (d, J = 8.5 Hz, 1H), 7.61 (d, J = 2.3 Hz, 1H), 7.47 (dd, J = 8.5, 2.4 Hz, 1H), 3.34 (s, 3H). 13 CNMR (101 MHz, DMSO) δ 160.6, 149.3, 146.7, 141.0, 139.1, 136.4, 136.0, 133.6, 133.5, 133.4, 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] Example 19:

[0124]

[0125] Following the synthetic procedure for 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 1,3-benzothiazole-6-sulfonyl chloride to give intermediate 19-a.

[0126]

[0127] Following the synthetic procedure for 1-b in Example 1, intermediate 19-a was deacetylated with concentrated hydrochloric acid to give intermediate 19-b.

[0128]

[0129] Following the synthetic procedure for 1-c in Example 1, intermediate 19-b was reacted with thiophosgene to give intermediate 19-c.

[0130]

[0131] Following the synthetic procedure for 1-d in Example 1, intermediate 19-c was reacted with 2-amino-5-methylsulfonebenzoic acid to give intermediate 19-d.

[0132] The target compound 19 was obtained following the synthetic procedure for the target compound 1 in Example 1, reacting intermediate 19-d with thiophosgene to give the target compound 19 in 29% yield. 1HNMR (400 MHz, DMSO) δ 10.34 (s, 1H), 9.62 (s, 1H), 8.62 (d, J = 1.6 Hz, 1H), 8.56 (d, J = 2.1 Hz, 1H), 8.36 (dd, J = 8.6, 2.1 Hz, 1H), 8.24 (d, J = 8.6 Hz, 1H), 7.95 - 7.84 (m, 2H), 7.62 (d, J = 8.5 Hz, 1H), 7.57 (d, J = 2.3 Hz, 1H), 7.41 (dd, J = 8.5, 2.3 Hz, 1H), 3.34 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 161.0, 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] Example 20:

[0134]

[0135] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with 4-carboxymethyl phenylsulfonyl chloride to give intermediate 20-a.

[0136]

[0137] Following the synthetic procedure of Example 1 for 1-b, intermediate 20-a was acetylated with thionyl chloride and methanol to give intermediate 20-b.

[0138]

[0139] Following the synthetic procedure of Example 1 for 1-c, intermediate 20-b was reacted with thiophosgene to give intermediate 20-c.

[0140]

[0141] Following the synthetic procedure of Example 1 for 1-d, intermediate 20-c was reacted with 2-amino-5-methylsulfonophenylboronic acid to give intermediate 20-d.

[0142] Following the synthetic procedure of Example 1 for 1, intermediate 20-d was reacted with thiophosgene to give target compound 20 in 37% yield. 1H NMR (400 MHz, DMSO) δ 10.44 (s, 1H), 8.57 (d, J = 2.0 Hz, 1H), 8.36 (dd, J = 8.6, 2.1 Hz, 1H), 8.11 (d, J = 8.4 Hz, 2H), 7.92 (d, J = 8.6 Hz, 1H), 7.85 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 8.5 Hz, 1H), 7.57 (d, J = 2.3 Hz, 1H), 7.44 (dd, J = 8.5, 2.3 Hz, 1H), 3.88 (s, 3H), 3.34 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 165.0, 160.6, 149.3, 146.7, 143.5, 139.0, 136.3, 133.8, 133.2, 132.7, 130.7, 130.7, 129.9, 128.1, 127.9, 127.1, 126.4, 121.0, 52.5, 43.3.

[0143] Example 21:

[0144]

[0145] Intermediate 21-a (150 mg, 0.27 mmol) was dissolved in 2 mL DMF, 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 reaction was allowed to proceed at room temperature for h, then washed with saturated brine 3 times, extracted with ethyl acetate, the organic phases were combined, concentrated to dryness, and column chromatography was performed to obtain intermediate 21-b as a white solid 128 mg, 57% yield.

[0146]

[0147] Intermediate 21-a (150 mg, 0.27 mmol) was dissolved in 2 mL DMF, 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 reaction was allowed to proceed at room temperature for h, then washed with saturated brine 3 times, extracted with ethyl acetate, the organic phases were combined, concentrated to dryness, and column chromatography was performed to obtain intermediate 21-b as a white solid 128 mg, 57% yield. 1H NMR (800 MHz, DMSO) δ 13.40 (s, 1H), 10.25 (s, 1H), 8.70 (s, 1H), 8.40 (s, 1H), 8.27 (d, J = 8.3 Hz, 1H), 7.98 - 7.89 (m, 3H), 7.78 (d, J = 7.3 Hz, 2H), 7.61 (d, J = 8.3 Hz, 1H), 7.53 (d, J = 8.0 Hz, 1H), 7.44 (s, 1H), 7.21 (d, J = 7.9 Hz, 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 = 11.4 Hz, 1H), 2.58 (d, J = 12.4 Hz, 1H), 2.07 (d, J = 7.6 Hz, 2H), 1.60 (s, 1H), 1.55 - 1.42 (m, 3H), 1.30 (d, J = 18.5 Hz, 2H). 13 C NMR (201 MHz, DMSO) δ 176.7, 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] Target compound 21 was obtained by reacting intermediate 21-b with thiophosgene according to the synthetic method of target compound 1 in Example 1, in a yield of 27%. 1H NMR (800 MHz, DMSO) δ 10.38 (s, 1H), 8.70 (s, 1H), 8.58 (s, 1H), 8.37 (d, J = 8.1 Hz, 1H), 7.98 (d, J = 7.6 Hz, 2H), 7.96 - 7.90 (m, 2H), 7.81 (d, J = 7.6 Hz, 2H), 7.67 (s, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.44 (d, J = 8.1 Hz, 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 = 12.0 Hz, 1H), 2.58 (d, J = 12.4 Hz, 1H), 2.07 (s, 2H), 1.60 (s, 1H), 1.56 - 1.42 (m, 3H), 1.30 (d, J = 18.1 Hz, 2H). 13 C NMR (201 MHz, DMSO) δ 172.8, 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] Example 22:

[0151]

[0152] Intermediate 22-b

[0153]

[0154] Intermediate 22-b was obtained by reacting intermediate 21-a with (3AS,4S,6AR)-N-[2-[2-(2- aminoethoxy)ethoxy]ethyl]hexahydro-2-oxo-1H-thieno[3,4-D]imidazole-4-pentanamide according to the synthetic procedure of Example 21 for 21-b. 1H NMR (800 MHz, DMSO) δ 13.39 (s, 1H), 10.22 (s, 1H), 8.75 (s, 1H), 8.40 (s, 1H), 8.27 (d, J = 8.3 Hz, 1H), 7.96 (d, J = 7.5 Hz, 2H), 7.82 (s, 1H), 7.78 (d, J = 7.5 Hz, 2H), 7.61 (d, J = 8.4 Hz, 1H), 7.53 (d, J = 8.1 Hz, 1H), 7.43 (s, 1H), 7.22 (d, J = 8.1 Hz, 1H), 6.39 (d, J = 43.9 Hz, 2H), 4.31 (s, 1H), 4.13 (s, 1H), 3.53 (d, J = 18.9 Hz, 6H), 3.41 (d, J = 24.3 Hz, 4H), 3.30 (s, 3H), 3.18 (s, 2H), 3.09 (s, 1H), 2.82 (d, J = 11.5 Hz, 1H), 2.58 (d, J = 12.4 Hz, 1H), 2.06 (s, 2H), 1.61 (s, 1H), 1.55 - 1.42 (m, 3H), 1.35 - 1.25 (m, 2H). 13 C NMR (201 MHz, DMSO) δ 177.2, 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] Target compound 22 was obtained by reacting intermediate 22-b with thiophosgene according to the synthetic method of target compound 1 in Example 1, in 23% yield. 1H NMR (400 MHz, DMSO) δ 10.37 (s, 1H), 8.74 (s, 1H), 8.57 (s, 1H), 8.36 (d, J = 8.3 Hz, 1H), 7.98 (d, J = 7.4 Hz, 2H), 7.93 (d, J = 8.2 Hz, 1H), 7.83 - 7.77 (m, 3H), 7.64 (d, J = 14.1 Hz, 2H), 7.43 (d, J = 8.2 Hz, 1H), 6.38 (d, J = 44.1 Hz, 2H), 4.30 (s, 1H), 4.12 (s, 1H), 3.52 (d, J = 19.2 Hz, 6H), 3.40 (d, J = 28.0 Hz, 4H), 3.35 (s, 3H), 3.17 (d, J = 3.1 Hz, 2H), 3.08 (s, 1H), 2.81 (d, J = 11.9 Hz, 1H), 2.57 (d, J = 12.4 Hz, 1H), 2.06 (t, J = 10.8 Hz, 2H), 1.60 (s, 1H), 1.55 - 1.41 (m, 3H), 1.35 - 1.24 (m, 2H). 13 C NMR (201 MHz, DMSO) δ 172.1, 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] Example 23:

[0158]

[0159] Following the procedure for the synthesis of 1-d in Example 1, intermediate 23-d was reacted with thiophosgene to give target compound 23 in 36% yield.

[0160] Target compound 23 was obtained following the procedure for the synthesis of target compound 1 in Example 1, reacting intermediate 23-d with thiophosgene to give target compound 23 in 36% yield. 1 H NMR (400 MHz, DMSO) δ 10.59 (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 = 7.6 Hz, 1H). 13C NMR (101 MHz, DMSO) δ 159.6, 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] Example 24:

[0162]

[0163]

[0164] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 3-amino-6-chloropyridine-2-carboxylic acid to give intermediate 24-d.

[0165] Target compound 24 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 24-d with thiophosgene to give target compound 24 in 32% yield. 1 H NMR (400 MHz, DMSO) δ 10.58 (s, 1H), 8.20 (d, J = 8.6 Hz, 1H), 7.98 (d, J = 8.6 Hz, 1H), 7.73 - 7.64 (m, 3H), 7.60 (d, J = 2.3 Hz, 1H), 7.47 - 7.38 (m, 2H), 7.32 (t, J = 7.6 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 159.6, 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] Example 25:

[0167]

[0168] Following the synthetic procedure of Example 1 for 1-d, intermediate 1-c was reacted with 2-amino-5-chloropyridine-3-carboxylic acid to give intermediate 25-d.

[0169] Target compound 25 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 25-d with thiophosgene to give target compound 25 in 32% yield. 1H NMR (400 MHz, DMSO) δ 10.60 (s, 1H), 9.04 (d, J = 2.7 Hz, 1H), 8.55 (d, J = 2.7 Hz, 1H), 7.75 - 7.58 (m, 4H), 7.47 - 7.39 (m, 2H), 7.32 (dd, J = 11.3, 4.0 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.9, 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] Example 26:

[0171]

[0172]

[0173] Following the procedure for the synthesis of 1-d in Example 1, intermediate 26-d was obtained by reacting intermediate 1-c with 3-aminoisonicotinic acid. Target compound 26 was obtained by reacting intermediate 26-d with thiophosgene following the procedure for the synthesis of target compound 1 in Example 1 in 43% yield. 1 H NMR (400 MHz, DMSO) δ 10.60 (s, 1H), 9.04 (d, J = 2.7 Hz, 1H), 8.55 (d, J = 2.7 Hz, 1H), 7.75 - 7.58 (m, 4H), 7.47 - 7.39 (m, 2H), 7.32 (dd, J = 11.3, 4.0 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.9, 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.

[0174] Example 27:

[0175]

[0176] Following the synthetic procedure of Example 1 for 1-d, intermediate 27-c was reacted with 3-amino-6-chloropyrazine-2-carboxylic acid to give intermediate 27-d.

[0177] Following the synthetic procedure of Example 1 for Compound 1, intermediate 27-d was reacted with thiophosgene to give Compound 27 in 26% yield. 1 H NMR (400 MHz, DMSO) δ 10.60 (s, 1H), 9.18 (s, 1H), 7.75 - 7.65 (m, 3H), 7.60 (d, J = 2.4 Hz, 1H), 7.48 - 7.37 (m, 2H), 7.34 (dd, J = 11.2, 4.0 Hz, 1H). 13 CNMR (101 MHz, DMSO) δ 159.5, 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] Example 28:

[0179]

[0180]

[0181] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with vinylsulfonyl chloride to give intermediate 28-a.

[0182]

[0183] Following the synthetic procedure of Example 1 for 1-b, intermediate 28-a was deacetylated with concentrated hydrochloric acid to give intermediate 28-b.

[0184]

[0185] Following the synthetic procedure of Example 1 for 1-c, intermediate 28-b was reacted with thiophosgene to give intermediate 28-c.

[0186]

[0187] Following the synthetic procedure of Example 1 for 1-d, intermediate 28-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 28-d.

[0188] Target compound 28 was obtained by reacting intermediate 28-d with thiophosgene according to the method described in Example 1 for the synthesis of target compound 1 in 33% yield. 1 HNMR (400 MHz, DMSO) δ 9.92 (s, 1H), 8.96 (s, 1H), 8.06 (d, J = 6.2 Hz, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.64 (d, J = 2.3 Hz, 1H), 7.41 (dd, J = 8.5, 2.3 Hz, 1H), 6.84 (dd, J = 16.4, 9.9 Hz, 1H), 6.07 (dd, J = 13.0, 11.5 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 159.6, 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] Example 29:

[0190]

[0191]

[0192] Intermediate 29-a was obtained by reacting 3-amino-4-chloroacetanilide with 5-chlorothiophene-2-sulfonyl chloride according to the method described in Example 1 for the synthesis of 1-a.

[0193]

[0194] Intermediate 29-b was obtained by removing the acetyl group of intermediate 29-a with concentrated hydrochloric acid according to the method described in Example 1 for the synthesis of 1-b.

[0195]

[0196] Intermediate 29-c was obtained by reacting intermediate 29-b with thiophosgene according to the method described in Example 1 for the synthesis of 1-c.

[0197]

[0198] Intermediate 29-d was obtained by reacting intermediate 29-c with 5-amino-2-chloropyridine-4-carboxylic acid according to the method described in Example 1 for the synthesis of 1-d.

[0199] Target compound 29 was obtained by reacting intermediate 29-d with thiophosgene according to the method described in Example 1 for the synthesis of target compound 1 in 33% yield. 1HNMR (400 MHz, DMSO) δ 10.63 (s, 1H), 8.96 (s, 1H), 8.05 (s, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 2.3 Hz, 1H), 7.46 (dd, J = 8.5, 2.4 Hz, 1H), 7.36 (d, J = 4.1 Hz, 1H), 7.23 (d, J = 4.1 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 159.6, 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] Example 30:

[0201]

[0202]

[0203] Following the synthetic procedure for 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2,3-dihydrobenzofuran-5-sulfonyl chloride to give intermediate 30-a.

[0204]

[0205] Following the synthetic procedure for 1-b in Example 1, intermediate 30-a was acetylated with concentrated hydrochloric acid to give intermediate 30-b.

[0206]

[0207] Following the synthetic procedure for 1-c in Example 1, intermediate 30-b was reacted with thiophosgene to give intermediate 30-c.

[0208]

[0209] Following the synthetic procedure for 1-d in Example 1, intermediate 30-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 30-d.

[0210] The target compound 30 was obtained following the synthetic procedure for target compound 1 in Example 1, reacting intermediate 30-d with thiophosgene to give target compound 30 in 37% yield. 1H NMR (400 MHz, DMSO) δ 9.94 (s, 1H), 8.96 (s, 1H), 8.05 (s, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.59 (d, J = 2.1 Hz, 2H), 7.50 (dd, J = 8.5, 1.8 Hz, 1H), 7.34 (dd, J = 8.5, 2.4 Hz, 1H), 6.86 (d, J = 8.5 Hz, 1H), 4.62 (t, J = 8.9 Hz, 2H), 3.19 (t, J = 8.8 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 163.4, 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] Example 31:

[0212]

[0213]

[0214] Following the synthetic procedure for 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 1-methyl-1H-pyrazole-4-sulfonyl chloride to give intermediate 31-a.

[0215]

[0216] Following the synthetic procedure for 1-b in Example 1, intermediate 31-a was deacetylated with concentrated hydrochloric acid to give intermediate 31-b.

[0217]

[0218] Following the synthetic procedure for 1-c in Example 1, intermediate 31-b was reacted with thiophosgene to give intermediate 31-c.

[0219]

[0220] Following the synthetic procedure for 1-d in Example 1, intermediate 31-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 31-d.

[0221] The target compound 31 was obtained following the synthetic procedure for the target compound 1 in Example 1, reacting intermediate 31-d with thiophosgene to give the target compound 31 in 25% yield. 1H NMR (400 MHz, DMSO) δ 9.96 (s, 1H), 8.97 (s, 1H), 8.22 (s, 1H), 8.06 (s, 1H), 7.73 - 7.63 (m, 2H), 7.59 (d, J = 2.3 Hz, 1H), 7.36 (dd, J = 8.5, 2.3 Hz, 1H), 3.85 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 159.7, 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] Example 32:

[0223]

[0224]

[0225] Following the synthetic procedure for 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 3,5-dimethylisoxazole-4-sulfonyl chloride to give intermediate 32-a.

[0226]

[0227] Following the synthetic procedure for 1-b in Example 1, intermediate 32-a was deacetylated with concentrated hydrochloric acid to give intermediate 32-b.

[0228]

[0229] Following the synthetic procedure for 1-c in Example 1, intermediate 32-b was reacted with thiophosgene to give intermediate 32-c.

[0230]

[0231] Following the synthetic procedure for 1-d in Example 1, intermediate 32-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 32-d.

[0232] The target compound 32 was obtained following the synthetic procedure for the target compound 1 in Example 1, reacting intermediate 32-d with thiophosgene to give the target compound 32 in 28% yield. 1 H NMR (400 MHz, DMSO) δ 9.96 (s, 1H), 8.97 (s, 1H), 8.22 (s, 1H), 8.06 (s, 1H), 7.73 - 7.63 (m, 2H), 7.59 (d, J = 2.3 Hz, 1H), 7.36 (dd, J = 8.5, 2.3 Hz, 1H), 3.85 (s, 3H).13 C NMR (101 MHz, DMSO) δ 173.1, 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] Example 33:

[0234]

[0235]

[0236] Following the synthetic procedure for 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 2-thiophenesulfonyl chloride to give intermediate 33-a.

[0237]

[0238] Following the synthetic procedure for 1-b in Example 1, intermediate 33-a was deacetylated with concentrated hydrochloric acid to give intermediate 33-b.

[0239]

[0240] Following the synthetic procedure for 1-c in Example 1, intermediate 33-b was reacted with thiophosgene to give intermediate 33-c.

[0241]

[0242] Following the synthetic procedure for 1-d in Example 1, intermediate 33-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 33-d.

[0243] The target compound 33 was obtained following the synthetic procedure for target compound 1 in Example 1, reacting intermediate 33-d with thiophosgene to give target compound 33 in 41% yield. 1 H NMR (400 MHz, DMSO) δ 10.42 (s, 1H), 8.96 (s, 1H), 8.05 (s, 1H), 7.94 (d, J = 4.8 Hz, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.60 (s, 1H), 7.47 (d, J = 3.6 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.13 (t, J = 3.9 Hz, 1H). 13C NMR (101 MHz, DMSO) δ 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.

[0244] Example 34:

[0245]

[0246]

[0247] Following the synthetic procedure of Example 1 for 1-d, intermediate 34-d was reacted with thiophosgene to give target compound 34 in 40% yield.

[0248] Target compound 34 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 34-d with thiophosgene to give target compound 34 in 40% yield. 1 H NMR (400 MHz, DMSO) δ 10.91 (s, 1 H), 8.95 (s, 1 H), 8.03 (s, 1 H), 7.78 - 7.66 (m, 2H), 7.62 (d, J = 2.3 Hz, 1 H), 7.45 (dd, J = 8.5, 2.3 Hz, 1 H), 7.25 (t, J = 9.1 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 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] Example 35:

[0250]

[0251] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with 2-chlorobenzenesulfonyl chloride to give intermediate 35-a.

[0252]

[0253] Following the synthetic procedure of Example 1 for 1-b, intermediate 35-a was deacetylated with concentrated hydrochloric acid to give intermediate 35-b.

[0254]

[0255] Following the synthetic procedure of Example 1 for 1-c, intermediate 35-b was reacted with thiophosgene to give intermediate 35-c.

[0256]

[0257] Following the synthetic procedure of Example 1 for 1-d, intermediate 35-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 35-d.

[0258] The target compound 35 was obtained following the synthetic procedure of Example 1 for compound 1, reacting intermediate 35-d with thiophosgene to give target compound 35 in 46% yield. 1 H NMR (400 MHz, DMSO) δ 10.46 (s, 1H), 8.94 (s, 1H), 8.03 (s, 1H), 7.87 (d, J = 7.7 Hz, 1H), 7.69 - 7.60 (m, 3H), 7.58 (d, J = 2.0 Hz, 1H), 7.48 (dd, J = 10.2, 4.2 Hz, 1H), 7.38 (dd, J = 8.5, 2.1 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 159.6, 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] Example 36:

[0260]

[0261] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with piperidine sulfonyl chloride to give intermediate 36-a.

[0262]

[0263] Following the synthetic procedure of Example 1 for 1-b, intermediate 36-a was deacetylated with concentrated hydrochloric acid to give intermediate 36-b.

[0264]

[0265] Following the synthetic procedure of Example 1 for 1-c, intermediate 36-b was reacted with thiophosgene to give intermediate 36-c.

[0266]

[0267] Following the synthetic procedure of Example 1 for 1-d, intermediate 36-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 36-d.

[0268] Target compound 36 was obtained following the synthetic procedure of Example 1 for target compound 1 by reacting intermediate 36-d with thiophosgene to give target compound 36 in 24% yield. 1 HNMR (400 MHz, DMSO) δ 9.55 (s, 1H), 8.95 (d, J = 0.6 Hz, 1H), 8.04 (d, J = 0.7 Hz, 1H), 7.72 (dd, J = 6.7, 5.5 Hz, 2H), 7.35 (dd, J = 8.5, 2.4 Hz, 1H), 3.23 - 3.02 (m, 4H), 1.54 - 1.34 (m, 6H). 13 C NMR (101 MHz, DMSO) δ 159.6, 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] Example 37:

[0270]

[0271] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with 2-trifluoromethylbenzenesulfonyl chloride to give intermediate 37-a.

[0272]

[0273] Following the synthetic procedure of Example 1 for 1-b, intermediate 37-a was deacetylated with concentrated hydrochloric acid to give intermediate 37-b.

[0274]

[0275] Following the synthetic procedure of Example 1 for 1-c, intermediate 37-b was reacted with thiophosgene to give intermediate 37-c.

[0276]

[0277] Following the synthetic procedure of Example 1 for 1-d, intermediate 37-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 37-d.

[0278] Target compound 37 was obtained following the synthetic procedure of Example 1 for target compound 1 by reacting intermediate 37-d with thiophosgene to give target compound 37 in 38%. 1H NMR (400 MHz, DMSO) δ 10.39 (s, 1H), 8.94 (s, 1H), 8.03 (s, 1H), 7.99 (d, J = 7.5 Hz, 1H), 7.91 (d, J = 7.7 Hz, 1H), 7.83 (dt, J = 20.6, 7.2 Hz, 2H), 7.63 (dd, J = 13.3, 5.4 Hz, 2H), 7.43 (dd, J = 8.5, 2.3 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 159.6, 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] Example 38:

[0280]

[0281] Following the synthetic procedure for 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with 3-pyridinesulfonyl chloride to give intermediate 38-a.

[0282]

[0283] Following the synthetic procedure for 1-b in Example 1, intermediate 38-a was deacetylated with concentrated hydrochloric acid to give intermediate 38-b.

[0284]

[0285] Following the synthetic procedure for 1-c in Example 1, intermediate 38-b was reacted with thiophosgene to give intermediate 38-c.

[0286]

[0287] Following the synthetic procedure for 1-d in Example 1, intermediate 38-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 38-d.

[0288] Target compound 38 was obtained following the synthetic procedure for target compound 1 in Example 1, reacting intermediate 38-d with thiophosgene to give target compound 38 in 26% yield. 1H NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 8.96 (d, J = 0.6 Hz, 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, 2.4 Hz, 1H). 13 CNMR (101 MHz, DMSO) δ 159.6, 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] Example 39:

[0290]

[0291] Following the synthetic procedure of Example 1 for 1-d, intermediate 19-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 40-d.

[0292] The target compound 39 was obtained following the synthetic procedure of Example 1 for Compound 1, reacting intermediate 39-d with thiophosgene to give target compound 39 in 38% yield. 1 H NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 8.96 (d, J = 0.6 Hz, 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, 2.4 Hz, 1H). 13 CNMR (101 MHz, DMSO) δ 159.6, 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] Example 40:

[0294]

[0295] Following the synthetic procedure of Example 1 for 1-d, intermediate 19-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 40-d.

[0296] Target compound 40 was obtained by reacting intermediate 40-d with thiophosgene according to the method described in Example 1 for the synthesis of target compound 1 in 30% yield. 1 HNMR (400 MHz, DMSO) δ 10.33 (s, 1H), 9.62 (s, 1H), 8.95 (d, J = 0.7 Hz, 1H), 8.61 (d, J = 1.6 Hz, 1H), 8.26 - 8.22 (m, 1H), 8.02 (d, J = 0.7 Hz, 1H), 7.87 (dd, J = 8.6, 1.9 Hz, 1H), 7.62 (d, J = 8.5 Hz, 1H), 7.55 (d, J = 2.3 Hz, 1H), 7.37 (dd, J = 8.5, 2.4 Hz, 1H). 13 CNMR (101 MHz, DMSO) δ 161.0, 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] Example 41:

[0298]

[0299] Intermediate 41-a was obtained by reacting 3-amino-4-chloroacetanilide with 6-chloropyridine-3-sulfonyl chloride according to the method described in Example 1 for the synthesis of 1-a.

[0300]

[0301] Intermediate 41-b was obtained by removing the acetyl group of intermediate 41-a with concentrated hydrochloric acid according to the method described in Example 1 for the synthesis of 1-b.

[0302]

[0303] Intermediate 41-c was obtained by reacting intermediate 41-b with thiophosgene according to the method described in Example 1 for the synthesis of 1-c.

[0304]

[0305] Intermediate 41-d was obtained by reacting intermediate 41-c with 5-amino-2-chloropyridine-4-carboxylic acid according to the method described in Example 1 for the synthesis of 1-d.

[0306] Target compound 41 was obtained by reacting intermediate 41-d with thiophosgene according to the method described in Example 1 for the synthesis of target compound 1 in 20% yield. 1H NMR (400 MHz, DMSO) δ 10.66 (s, 1H), 8.97 (d, J = 0.6 Hz, 1H), 8.68 - 8.66 (m, 1H), 8.09 (dd, J = 8.4, 2.6 Hz, 1H), 8.05 (d, J = 0.6 Hz, 1H), 7.76 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 8.5 Hz, 1H), 7.58 (d, J = 2.3 Hz, 1H), 7.45 (dd, J = 8.5, 2.4 Hz, 1H). 13 CNMR (101 MHz, DMSO) δ 159.6, 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] Example 42:

[0308]

[0309] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with (E)-2-styrene-1-sulfonyl chloride to give intermediate 42-a.

[0310]

[0311] Following the synthetic procedure of Example 1 for 1-b, intermediate 42-a was deacetylated with concentrated hydrochloric acid to give intermediate 42-b.

[0312]

[0313] Following the synthetic procedure of Example 1 for 1-c, intermediate 42-b was reacted with thiophosgene to give intermediate 42-c.

[0314]

[0315] Following the synthetic procedure of Example 1 for 1-d, intermediate 42-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 42-d.

[0316] The target compound 42 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 42-d with thiophosgene to give target compound 42 in 38% yield. 1HNMR (400 MHz, DMSO) δ 9.92 (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 = 15.4 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 159.6, 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] Example 43:

[0318]

[0319] Following the synthetic procedure for 1-a in Example 1, 3-amino-4-chloroacetanilide was reacted with N,N-dimethylsulfonyl chloride to give intermediate 43-a.

[0320]

[0321] Following the synthetic procedure for 1-b in Example 1, intermediate 43-a was deacetylated with concentrated hydrochloric acid to give intermediate 43-b.

[0322]

[0323] Following the synthetic procedure for 1-c in Example 1, intermediate 43-b was reacted with thiophosgene to give intermediate 43-c.

[0324]

[0325] Following the synthetic procedure for 1-d in Example 1, intermediate 43-c was reacted with 5-amino-2-chloropyridine-4-carboxylic acid to give intermediate 43-d.

[0326] The target compound 43 was obtained following the synthetic procedure for target compound 1 in Example 1, reacting intermediate 43-d with thiophosgene to give target compound 43 in 25% yield. 1 H NMR (400 MHz, DMSO) δ 9.61 (s, 1H), 8.96 (s, 1H), 8.05 (s, 1H), 7.75 (d, J = 2.3 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.38 (dd, J = 8.5, 2.4 Hz, 1H), 2.73 (s, 6H). 13CNMR (101 MHz, DMSO) δ 159.7, 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] Example 44:

[0328]

[0329] Following the procedure of Example 1 for the synthesis of 1-d, intermediate 44-d was reacted with thiophosgene to give the target compound 44 in 40% yield. 1 HNMR (400 MHz, DMSO) δ 10.92 (s, 1H), 9.07 (s, 1H), 8.74 (d, J = 5.1 Hz, 1H), 7.97 (d, J = 5.1 Hz, 1H), 7.78 - 7.66 (m, 2H), 7.64 (d, J = 2.3 Hz, 1H), 7.48 (dd, J = 8.5, 2.4 Hz, 1H), 7.26 (t, J = 9.1 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 160.6, 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] Example 45:

[0331]

[0332] Following the procedure of Example 1 for the synthesis of 1-a, 3-amino-4- chloroacetanilide was reacted with 2,4-difluorobenzenesulfonyl chloride to give intermediate 45-a.

[0333]

[0334] Following the procedure of Example 1 for the synthesis of 1-b, intermediate 45-a was deacetylated with concentrated hydrochloric acid to give intermediate 45-b.

[0335]

[0336] Following the synthetic procedure of Example 1 for 1-c, intermediate 45-b was reacted with thiophosgene to give intermediate 45-c.

[0337]

[0338] Following the synthetic procedure of Example 1 for 1-d, intermediate 45-c was reacted with 3-aminoisonicotinic acid to give intermediate 45-d. Target compound 45 was obtained following the synthetic procedure of Example 1 for target compound 1, reacting intermediate 45-d with thiophosgene to give target compound 45 in 42% yield. 1 H NMR (400 MHz, CDC13) δ 9.13 (s, 1H), 8.77 (d, J = 5.2 Hz, 1H), 8.03 - 7.98 (m, 1H), 7.83 (td, J = 8.7, 6.1 Hz, 1H), 7.68 - 7.61 (m, 2H), 7.49 (d, J = 8.5 Hz, 1H), 7.03 - 6.91 (m, 3H). 13 C NMR (101 MHz, CDC13) δ 167.9, 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] Example 46:

[0340]

[0341] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with 2,5-difluorobenzenesulfonyl chloride to give intermediate 46-a.

[0342]

[0343] Following the synthetic procedure of Example 1 for 1-b, intermediate 46-a was deacetylated with concentrated hydrochloric acid to give intermediate 46-b.

[0344]

[0345] Following the synthetic procedure of Example 1 for 1-c, intermediate 46-b was reacted with thiophosgene to give intermediate 46-c.

[0346]

[0347] Following the synthetic method of 1-d in Example 1, intermediate 46-c was reacted with 3-aminoisocarboxylic acid to obtain intermediate 46-d. Target compound 46 was obtained by reacting intermediate 46-d with phosgene according to the synthetic method of target compound 1 in Example 1, with 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] Example 47:

[0349]

[0350] Following 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] Following the synthesis method of 1-b in Example 1, intermediate 47-a was deacetylated with concentrated hydrochloric acid to obtain intermediate 47-b.

[0353]

[0354] Following the synthesis method of 1-c in Example 1, intermediate 47-b reacts with sulfur phosgene to obtain intermediate 47-c.

[0355]

[0356] Following the synthetic procedure of Example 1 for 1-d, intermediate 47-c was reacted with 3-aminoisonicotinic acid to give intermediate 47-d. Target compound 47 was obtained following the synthetic procedure of Example 1 for target compound 1 by reacting intermediate 47-d with thiophosgene in 35% yield. 1 HNMR (400 MHz, DMSO) δ 10.83 (s, 1H), 9.08 (s, 1H), 8.75 (d, J = 5.1 Hz, 1H), 7.97 (d, J = 5.1 Hz, 1H), 7.87 (td, J = 10.0, 6.5 Hz, 1H), 7.76 - 7.65 (m, 2H), 7.55 (d, J = 2.1 Hz, 1H), 7.48 (dd, J = 8.5, 2.1 Hz, 1H). 13 C NMR (101 MHz, DMSO) δ 160.6, 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, 141.1, 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] Example 48:

[0358]

[0359] Following the synthetic procedure of Example 1 for 1-a, 3-amino-4-chloroacetanilide was reacted with 2-fluoro-4-nitrobenzenesulfonyl chloride to give intermediate 48-a.

[0360]

[0361] Following the synthetic procedure of Example 1 for 1-b, intermediate 48-a was deacetylated with concentrated hydrochloric acid to give intermediate 48-b.

[0362]

[0363] Following the synthetic procedure of Example 1 for 1-c, intermediate 48-b was reacted with thiophosgene to give intermediate 48-c.

[0364]

[0365] Following the synthetic procedure of Example 1 for 1-d, intermediate 48-c was reacted with 3-aminoisonicotinic acid to give intermediate 48-d.

[0366] Target compound 48 was obtained by reacting intermediate 48-d with thiophosgene according to the synthetic method of target compound 1 in example 1, in a yield of 46%. 1 H NMR (400 MHz, CDC13) δ 9.16 (s, 1H), 8.80 (d, J = 5.2 Hz, 1H), 8.14 - 8.01 (m, 4H), 7.67 (d, J = 2.4 Hz, 1H), 7.56 (s, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.04 (dd, J = 8.5, 2.4 Hz, 1H). 13 C NMR (101 MHz, CDC13) δ 160.7, 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] Example 49:

[0368] In vitro antagonistic effect of sulfonamide compounds on NOD1

[0369] The in vitro screening system of NOD1 used in the present application is based on HEK-Blue hNOD1 (human NOD1) cell line and HEK-Blue mNOD1 (murine NOD1) cell line with high expression of NOD1, and uses secreted alkaline phosphatase (SEAP) as a reporter system. iE-DAP is used as an agonist of NOD1, and antagonists are screened. In such screening cells, the SEAP reporter gene is integrated into a plasmid vector of the NF-κB promoter, so that the activation or antagonism of NOD1 signaling pathway by a compound can be detected by evaluating the secretion level of alkaline phosphatase.

[0370] Experimental method:

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

[0372] Inhibitory Percentage (%) = [(C-T) / C] x 100. Wherein 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 values: the compound concentration has multiple gradients of 10 μM, 1 μM, 0.5 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.001 μM and 0.0001 μM. The operation method is the same as before. The inhibitory percentage of each concentration = [(C-T) / C] x 100, the antagonist and concentration curve are fitted, and the IC 50 .

[0374] The activity screening results of the sulfamide compounds are shown in Table 1.

[0375] Table 1 Activity screening results of sulfamide compounds

[0376]

[0377]

[0378] ND: not detected

[0379] From the above screening results, it can be seen that the sulfamide compounds have certain inhibitory effect on the NOD1 signaling pathway of two species (human and mouse).

[0380] Example 50

[0381] From the four compounds preferred in Example 49, the selectivity of the four compounds was studied, and the inhibitory effect of the four compounds on NOD2 and downstream node proteins RIP2 and Toll-like receptor (hTLR4) was evaluated. The experimental method refers to Example 49. The results are shown in Table 2.

[0382] Table 2 Selectivity of sulfamide compounds

[0383]

[0384]

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

[0386] Example 51

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

[0388] NOD1 signaling pathway is activated, it can induce the secretion of inflammatory cytokines such as IL-6, TNF-a, etc. In this experiment, we evaluated the effect of compound 17 on the secretion of inflammatory cytokines mediated by NOD1 signaling pathway by measuring the transcription level (mRNA) of IL-6 and TNF-a by ELISA. Figure 1 As can be seen from Table 7, the mRNA levels of IL-6 and TNF-a 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-a to near the initial state at 0.1 μM. This shows 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 NOD1 specific agonist C12-iE-DAP (1 μg / mL) was added to activate the NOD1 signaling pathway to produce inflammatory factors for 2 h, and finally the mRNA levels of IL-6 and TNF-a were measured 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 NOD1 specific agonist C12-iE-DAP (20 μg / mL) was added to activate the NOD1 signaling pathway to produce inflammatory factors for 2 h, and finally the mRNA levels of IL-6 and TNF-a were measured by q-RT-PCR. Data are expressed as mean ± SD (n = 3). Compared with cells stimulated only with C12-iE-DAP, (**) p < 0.01, (*** ) p < 0.001.

[0391] Example 52

[0392] Compound 17 antagonizes the secretion of inflammatory cytokines induced by H. pylori.

[0393] H. pylori colonizes the stomach, activates the NOD1 signaling pathway of the stomach cells, produces a large amount of inflammatory factors, and finally develops 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 shown in Table 8. Figure 2 Figure 2 ​A shows that the mRNA levels of IL-6 and CXCL8 are rapidly increased after THP-1 cells are stimulated by C12-iE-DAP or H. pylori, while Compound 17 at a concentration of 5 μM can inhibit the increase of IL-6 mRNA level caused by H. pylori stimulation to the initial level before stimulation, and also has a certain inhibitory effect on the expression level of CXCL8. Figure 2 B shows that Compound 17 at a concentration of 5 μM has an inhibition effect of nearly 50% on the mRNA level of CXCL1 in BMDM cells, and also has a certain inhibitory effect on the mRNA level of TNF-α in a dose-dependent manner. It can be seen that Compound 17 also has a good antagonistic effect on the secretion of inflammatory cytokines induced by H. pylori.

[0394] Experimental method:

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

[0396] Example 53

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

[0398] To evaluate the anti-inflammatory effect of Compound 17 in vivo, we established a mouse inflammatory model caused by the activation of the NOD1 signaling pathway by C12-iE-DAP stimulation. After the mice were injected intraperitoneally with C12-iE-DAP, the content of KC (the same homolog as IL-8 in rodents) in the serum was significantly higher than that of the control group, indicating that the inflammatory model was successfully established. The selective NOD1 antagonist Compound A reported in the literature was selected as a positive control drug, and its structure is shown as follows. Figure 3 Figure 4 ​As shown, compound 17 inhibited the serum KC value caused by C12-iE-DAP stimulation in mice in a dose-dependent manner by oral administration, and the anti-inflammatory effect of compound 17 at the same dose was comparable to that of the positive compound Compound A.

[0399] Experimental method:

[0400] Compound 17 was administered orally at a dose of 12.5, 25 and 50 mg / kg, respectively, and 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, and 15 min later, C12-iE-DAP (0.1 mg / kg) was injected intraperitoneally. The blank control group was injected intraperitoneally with the same volume of solvent. Blood was taken 2 h later, and the serum KC value of each group was determined by ELISF method. The data are expressed as mean ± SD (n = 5). Compared with the group administered with C12-iE-DAP only, ns indicates no difference, (*) p < 0.05, (**) p < 0.01, and (***) p < 0.001.

[0401] Example 54

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

[0403] To further investigate whether compound 17 can also inhibit H. pylori-induced gastric inflammation in mice, we first established an animal model of H. pylori-infected mouse stomach. 1 × 10 9 CFU / mL of bacteria solution was administered orally at a dose of 0.2 mL per 20 g of mouse body weight, and 3 days later, the mouse stomach tissue was taken, homogenized, and centrifuged to obtain the supernatant for KC content determination by Elisa method. Compound 17 and the positive control Compound A were administered orally at five time points before and after modeling. The experimental results are shown in Figure 5 As shown, the KC value of the stomach tissue after H. pylori infection increased significantly (DMSO group), and the KC value of the positive control Compound A (NOD1 antagonist reported in the literature) group decreased relative to the DMSO group. Compound 17 at a dose of 12.5 mg / kg had a better inhibitory effect on the KC value of the stomach tissue than Compound A, and the inhibitory effect on KC increased with the increase of the dose of compound 17, reaching the maximum inhibitory rate when administered orally at a dose of 50 mg / kg. The above results show that oral administration of compound 17 can inhibit H. pylori-induced gastric inflammation in mice.

[0404] Experimental method:

[0405] The administration group was orally administered with compound 17 (12.5, 25, 50 mg / kg) and positive drug Compound A (25 mg / kg) at 24 h and 2 h before modeling, and 0.2 mL of H. pylori (HP) bacterial solution (1 x 10 9 CFU) / 20 g was orally administered to induce gastric inflammation. The mice were administered in the same way for 3 times at 24 h, 48 h, and 72 h after modeling, and the mice were sacrificed at 2 h after the last administration, and the gastric tissue was measured for KC value. The negative control group (DMSO group) was modeled and administered with blank solvent, and the blank control group (Ctrl group) was not modeled and administered with blank solvent. The data is expressed as mean ± SD (n = 4), (*) p < 0.05, (**) p < 0.01, (***) p < 0.001 compared with the DMSO group.

Claims

1. A sulfonamide compound or its pharmaceutical salt, with the following specific structure: 。 2. The use of the sulfonamide compound as described in any one of claims 1 in the preparation of a medicament for the prevention or treatment of inflammatory diseases, wherein the inflammatory disease is gastritis caused by Helicobacter pylori infection.

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