Aminobenzenesulfonamide derivatives and uses thereof

Abstract

The application discloses an amino benzene sulfonamide derivative and application thereof. The structural general formula of the amino benzene sulfonamide derivative is shown in the following formula. The amino benzene sulfonamide derivative has good pepsin inhibitory activity, can effectively block the adhesion and reactivation of pepsin in the pharyngeal mucosa, and has super high selectivity in the internal proteinase family compared with other non-specific protease inhibitors, thereby significantly reducing the off-target risk of normal physiological proteases in the human body. In addition, the compound is particularly suitable for local spray administration in the throat, can maintain a high concentration in the lesion site, and overcomes the defect of traditional oral antacid drugs in the insufficient effect on non-acid reflux, thereby providing a new solution for the treatment of throat reflux diseases.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to an aminobenzenesulfonamide derivative and its applications. Background Technology

[0002] Laryngopharyngeal reflux (LPR) has long been considered one of the most complex and challenging problems to diagnose and treat in the field of otolaryngology. As research on LPR has shifted from a simple phenomenological study of gastric acid reflux to a mechanistic analysis of the components of the refluxate, it has become increasingly clear that its pathogenesis is not solely caused by gastric acid, but rather by the combined action of gastric acid and pepsin. In some non-acidic reflux environments, pepsin even plays a dominant damaging role. Common mechanisms include abnormal deposition of pepsin in the pharyngeal mucosa, reactivation in an acidic microenvironment, direct proteolytic action on mucosal cells, induction of inflammatory cytokine release, and disruption of epithelial barrier function.

[0003] Laryngopharyngeal reflux disease (LPR) is another serious upper gastrointestinal disorder following gastroesophageal reflux disease (GERD). Studies have shown that pepsin is not only a marker of reflux events but also a key effector molecule leading to tissue damage in the pharynx. Inhibiting pepsin activity or preventing its attachment and activation in the pharynx can directly reduce mucosal damage and thus improve symptoms in LPR patients. Although pepsin is inactive in a non-acidic environment (pH>4), it can still attach to cell surfaces and rapidly regain activity upon re-exposure to acidic stimuli, causing "secondary damage." Current treatment strategies mainly focus on proton pump inhibitors (PPIs) to suppress gastric acid secretion, aiming to inactivate pepsin by increasing pH. However, while various drugs for relieving LPR symptoms have been developed and used clinically, simple acid suppression therapy has limited efficacy for LPR caused by non-acidic or weakly acidic reflux, and symptoms in some patients cannot be fundamentally relieved.

[0004] Current clinical treatments for pepsin primarily rely on physical blocking (such as alginate) or broad-spectrum acid suppression, lacking highly selective and high-affinity small-molecule inhibitors. Traditional protease inhibitor development has largely focused on the virology or oncology fields. For example, in HIV treatment, HIV aspartic protease inhibitors (such as Amprenavir) have demonstrated excellent structural design. Amprenavir, as a highly effective HIV protease inhibitor, has a core structure that tightly fills the enzyme's binding pocket, mimicking the substrate transition state and forming a crucial hydrogen bond network with catalytic aspartic residues, thereby potently inhibiting enzyme activity. Notably, pepsin and HIV protease belong to the same aspartic protease family, sharing significant similarities in catalytic mechanisms and active site structures (both contain two key aspartic residues as catalytic centers). However, directly using Amprenavir for LPR treatment has limitations: its pharmacokinetic characteristics are designed for systemic absorption, and its selectivity for pepsin has not been optimized, potentially leading to off-target effects or insufficient local concentrations. Because its original structure was not customized for the unique substrate-specific pocket (S1-S4 subsites) of pepsin, and it lacks formulation adaptability for local throat administration, it cannot be directly used as an ideal drug for treating LPR.

[0005] Despite the significant success of ampunavir and its derivatives in the antiviral field, the application of its scaffold to the development of pepsin inhibitors remains a gap. Existing pepsin inhibition strategies lack highly selective small-molecule drugs based on rational drug design. Therefore, using the HIV protease inhibitor ampunavir as a lead compound scaffold, leveraging its core hydroxyethylamine transition state analog structural features, and targeting structural differences in the active pocket structure of pepsin for structural modification and optimization, to design a series of aminobenzenesulfonamide derivatives with high affinity, high selectivity, and suitability for local throat administration has become a highly promising research direction in this field. Developing such novel pepsin inhibitors based on the ampunavir scaffold holds the promise of overcoming the bottlenecks of existing LPR treatments, providing a new treatment strategy with a clear mechanism and definite efficacy for laryngopharyngeal reflux disease. Summary of the Invention

[0006] To address the shortcomings of the prior art, the present invention provides an aminobenzenesulfonamide derivative and its use in medicaments for the prevention and / or treatment of pepsin-related diseases (especially laryngopharyngeal reflux disease).

[0007] This invention relates to aminobenzenesulfonamide derivatives, which are rationally designed based on the homology of the aspartic protease family catalytic mechanism using the HIV protease inhibitor amprenavir as a lead backbone. This series of compounds exhibits excellent pepsin inhibitory activity, effectively blocking the attachment and reactivation of pepsin on the pharyngeal mucosa. Furthermore, compared to other non-specific protease inhibitors, they demonstrate extremely high selectivity within the protease family, significantly reducing the off-target risk of normal physiological proteases. In addition, the compounds of this invention are particularly suitable for local spray administration to the throat, maintaining high concentrations at the lesion site, overcoming the shortcomings of traditional oral acid-suppressing drugs in treating non-acid reflux, and providing a new solution for the treatment of laryngeal reflux disease.

[0008] The aminobenzenesulfonamide derivative of this invention has the following general structural formula:

[0009] ;

[0010] In the general formula:

[0011] X is O, S, or N;

[0012] R1 is selected from , , , , m can be 1, 2, 3 or 4;

[0013] R2 is selected from , , , , , , , , , , , .

[0014] R3 is selected from hydrogen, deuterium, halogen, methyl, hydroxyl, methoxy, ethoxy, hydroxymethyl, amino, nitro. , , , , , , , , , , , , , , , m can be 1, 2, 3 or 4.

[0015] Ring A is selected from one of the following: benzene ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring, 1,3,5-triazine ring, 1,2,4-triazine ring, and 1,2,3-triazine ring.

[0016] Furthermore, the aminobenzenesulfonamide derivative is selected from compounds with the following structures:

[0017] ;

[0018] ;

[0019] ;

[0020] ;

[0021] ;

[0022] .

[0023] The present invention relates to the application of aminobenzenesulfonamide derivatives in the preparation of pharmaceutical formulations.

[0024] The pharmaceutical preparation is a pharmaceutical preparation for the prevention and / or treatment of diseases related to pepsin.

[0025] The pepsin-related diseases include, but are not limited to, laryngopharyngeal reflux diseases (LPR), such as reflux laryngitis, laryngopharyngeal reflux disease, chronic cough caused by reflux, vocal cord nodules, vocal cord polyps, foreign body sensation in the throat, and related upper respiratory tract mucosal damage.

[0026] The present invention also provides a pharmaceutical composition comprising the aminobenzenesulfonamide derivative or a pharmaceutically acceptable salt, cocrystal, or solvate thereof.

[0027] The pharmaceutical composition also includes pharmaceutically acceptable excipients and carriers.

[0028] Preferably, the pharmaceutical composition is formulated as a spray (including solution spray, suspension spray, or powder spray) to meet the local drug delivery needs of the throat. Of course, depending on the treatment requirements, the pharmaceutical composition may also be prepared as tablets, capsules, powders, granules, syrups, solutions, oral liquids, mouthwashes, gels, or patches, etc.

[0029] The pharmaceutical composition is administered via throat spray, oral administration, sublingual administration, or topical application.

[0030] Specifically, the aminobenzenesulfonamide derivative can also exist in the drug as its solvate or as a pharmaceutically acceptable salt thereof. The term "pharmaceutically acceptable salt" refers to a salt formed by the compound with an acid or base that is suitable for use as a drug, including inorganic and organic salts. A preferred type of salt in this invention is a salt formed by the aminobenzenesulfonamide derivative and an acid. Suitable acids for salt formation include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, citric acid, tartaric acid, fumaric acid, maleic acid, methanesulfonic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

[0031] Specifically, the drug also includes a pharmaceutically acceptable carrier. A "pharmaceutically acceptable" component is a substance suitable for use in humans or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio. A "pharmaceutically acceptable carrier" is a pharmaceutically or food-grade solvent, suspending agent, or excipient used to deliver the compounds of the present invention to animals or humans. The carrier can be liquid or solid.

[0032] Specifically for spray formulations, pharmaceutically acceptable carriers include, but are not limited to: solvents or co-solvents such as purified water, ethanol, propylene glycol, glycerin, and polyethylene glycol (PEG); buffer salts (such as phosphate buffer and citrate buffer) to adjust the pH to a suitable range for the throat mucosa; surfactants (such as polysorbate and poloxamer) to increase the solubility or stability of poorly soluble drugs; and flavoring agents (such as menthol and sweeteners) and humectants.

[0033] For other dosage forms, the carrier may also include sugars (such as lactose, glucose, and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose, and methyl cellulose), tragacanth powder, malt, gelatin, talc, solid lubricants (such as stearic acid and magnesium stearate), vegetable oils, emulsifiers, wetting agents, colorants, stabilizers, antioxidants, preservatives, pyrogen-free water, isotonic salt solutions, etc.; the carrier can improve the stability, activity, and bioavailability of the formulation as needed, especially enhancing the retention time and permeability of the drug in the pharyngeal mucosa.

[0034] The beneficial effects of this invention are reflected in:

[0035] This invention is based on the structural framework of the HIV protease inhibitor Amprenavir. Taking advantage of the fact that pepsin and HIV protease belong to the same aspartic protease family and have similar catalytic mechanisms (both contain a bis-aspartic catalytic center), and based on the differences in the amino acid residues in their active pockets, the core structure of Amprenavir was modified and optimized using a structure-based drug design (SBDD) method. A series of aminobenzenesulfonamide derivatives that are expected to have good inhibitory activity and selectivity against pepsin were designed.

[0036] Chemical synthesis and further bioactivity tests (enzymatic inhibition experiments) showed that the aminobenzenesulfonamide derivatives retained the key sulfonamide group and the hydroxyethylamine structural features that mimic the transition state, while introducing side chain groups adapted to the pepsin-specific substrate binding pocket (S1-S4 subsites), which made them exhibit excellent inhibitory activity against pepsin (low nanomolar IC50), and within the aspartic protease family (such as against human renin, cathepsin D, etc.).

[0037] Further biological experiments confirmed that some compounds can effectively inhibit the proteolytic damage of pepsin to pharyngeal epithelial cells, reduce the release of inflammatory factors, and significantly improve laryngeal mucosal edema and inflammatory infiltration in animal models of laryngopharyngeal reflux disease (LPR). Furthermore, by formulating them into a throat spray, high-concentration targeted delivery to the lesion site can be achieved, rapidly neutralizing or inhibiting the activity of pepsin refluxed to the pharynx. This overcomes the limitations of traditional oral proton pump inhibitors (PPIs) in treating non-acidic reflux and systemic administration, demonstrating the great potential of this series of compounds in treating pepsin-mediated laryngopharyngeal reflux disease. Detailed Implementation

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0039] In all embodiments of the present invention 1 The H NMR spectra were obtained using a 600M superconducting nuclear magnetic resonance spectrometer (AVANCE NEO600), and chemical shifts were expressed in ppm using a tetramethylsilane internal standard (0.00 ppm). 1 H NMR representation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If the coupling constant is provided, the unit is Hz.

[0040] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15~0.2 mm, and the diameter of the thin-layer chromatography separation and purification products is 0.4~0.5 mm.

[0041] Column chromatography used Yantai Huanghai silica gel 200~300 mesh as the carrier.

[0042] Example 1:

[0043]

[0044] The synthesis route is shown below:

[0045]

[0046] Step 1: Synthesis of (2S,3R)-tert-butyl(1-(isobutylamino)-3-phenylpropane-2-yl)carbamate

[0047] (2S,3S)-2-benzyl-3-((tert-Butoxycarbonyl)aminomethyl)ethylene oxide (6.2 g, 1 equiv) was placed in a round-bottom flask, and isobutylamine (8.6 g, 5 equiv) was added to it using 80-120 mL of ethanol as solvent. The mixture was stirred at 60 °C for 6 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solvent was evaporated, and the product was extracted and subjected to column chromatography to obtain the target compound (6.8 g, white solid, yield 86.07%).

[0048] Step 2: Synthesis of (2S,3R)-tert-butyl((2-hydroxy-3-((4-nitrophenyl)sulfonyl(isobutyl)amino)-1-phenylpropyl)carbamate

[0049] (2S,3R)-tert-butyl(1-(isobutylamino)-3-phenylpropane-2-yl)carbamate (1.8 g, 1 equiv), 4-nitrobenzenesulfonyl chloride (1.78 g, 1.5 equiv), and triethylamine (1.62 g, 3 equiv) were placed in a round-bottom flask with 30-50 mL of dichloromethane as solvent. The mixture was stirred at room temperature for 6 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solvent was evaporated, and the product was extracted and subjected to column chromatography to obtain the target compound (2.1 g, yellow solid, yield 82.4%).

[0050] Step 3: Synthesis of (2S,3R)-tert-butyl((1-acetoxy-3-((4-nitrophenyl)sulfonyl(isobutyl)amino)-2-phenylpropyl)carbamate

[0051] (2S,3R)-tert-butyl((2-hydroxy-3-((4-nitrophenyl)sulfonyl(isobutyl)amino)-1-phenylpropyl)carbamate (2.3 g, 1 equiv) and 4-dimethylaminopyridine (1.57 g, 3 equiv) were placed in a round-bottom flask. 30-50 mL of dichloromethane was used as the solvent, and the mixture was stirred at 0°C for 5 min. 10-20 mL of acetic anhydride was added dropwise, and the mixture was stirred at room temperature for 1 h. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the mixture was neutralized with saturated sodium carbonate, and the target compound (2.1 g, white solid, yield 72.3%) was obtained by extraction and column chromatography.

[0052] Step 4: Synthesis of (3S)-tetrahydrofuran-3-yl-glycyl-O-acetyl-L-seryl-N-(4-nitrobenzenesulfonyl)-L-leucamide

[0053] (2S,3R)-tert-butyl((1-acetoxy-3-((4-nitrophenyl)sulfonyl(isobutyl)amino)-2-phenylpropyl)carbamate (2 g, 1 equiv) was placed in a round-bottom flask. Using 10-20 mL of dichloromethane as solvent, 2 mL of dichloropyridine and 2 mL of trifluoromethanesulfonic anhydride were added dropwise. The mixture was stirred at 0 °C for 10 min. Then, (S)-3-aminotetrahydrofuran (0.62 g, 2 equiv) and triethylamine (1.1 g, 3 equiv) were added. The mixture was stirred at room temperature for 12 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the mixture was evaporated to dryness, and the target compound (1.4 g, white solid, yield 66.6%) was obtained by extraction and column chromatography.

[0054] Step 5: Synthesis of (S)-3-aminotetrahydrofuran-glycyl-L-phenylalanyl-O-acetyl-L-seryl-N-(4-aminobenzenesulfonyl)-L-leucamide

[0055] (3S)-tetrahydrofuran-3-yl-glycyl-O-acetyl-L-seryl-N-(4-nitrobenzenesulfonyl)-L-leucamide (1 g, 1 equiv) was placed in a round-bottom flask. Palladium on carbon (18 mg, 0.1 equiv) was added as solvent in 10-20 mL of tetrahydrofuran. The mixture was stirred for 10 h under H2 conditions, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the filtrate was filtered and evaporated to dryness. The solution was then extracted and subjected to column chromatography to obtain the target compound (0.682 g, white solid, yield 72.01%).

[0056] Step 6: Synthesis of (S)-3-aminotetrahydrofuran-glycyl-L-phenylalanyl-L-seryl-N-(4-aminobenzenesulfonyl)-L-leucamide

[0057] (S)-3-aminotetrahydrofuran-glycyl-L-phenylalanyl-O-acetyl-L-seryl-N-(4-aminobenzenesulfonyl)-L-leucamide (200 mg, 1 equiv) was placed in a round-bottom flask, and potassium carbonate (164 mg, 3 equiv) was added to the flask using 5-10 mL of methanol as solvent. The mixture was stirred at room temperature for 2 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was completed, the target compound (120 mg, white solid, yield 63.4%) was obtained by extraction and column chromatography.

[0058] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.55 (d, J = 7.5 Hz, 2H), 7.26 (t,J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.58 (d,J = 7.5 Hz, 2H), 6.46 (br s, 2H), 6.06 (br s, 2H), 5.37 (s, 1H), 4.10-3.84(m, 5H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.19-1.95 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0059] Example 2:

[0060]

[0061] The preparation steps were the same as in Example 1, except that (S)-3-aminotetrahydrofuran was replaced with (S)-3-hydroxytetrahydrofuran, while other conditions remained unchanged.

[0062] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.38 (d, J = 7.5 Hz, 2H), 8.05 (d,J = 7.5 Hz, 2H), 7.67 (br s, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m,6H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.21-1.96 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0063] Example 3:

[0064]

[0065] The preparation steps are the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride is replaced with 4-methylbenzenesulfonyl chloride, and other conditions remain unchanged.

[0066] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.67 (br s, 1H), 7.65 (d, J = 7.5Hz, 2H), 7.45 (d, J = 7.5 Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m,6H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.43 (s, 3H), 2.21-1.96 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0067] Example 4:

[0068]

[0069] The preparation steps were the same as in Example 1, except that (S)-3-aminotetrahydrofuran was replaced with 3-amino-1-propanol, and other conditions remained unchanged.

[0070] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.55 (d, J = 7.5 Hz, 2H), 7.26 (t,J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.58 (d,J = 7.5 Hz, 2H), 6.46 (br s, 1H), 6.06 (br s, 2H), 6.02 (br s, 1H), 5.37 (s,1H), 4.39 (s, 1H), 4.07 (m, 1H), 3.88 (m, 1H), 3.42-3.38 (m, 6H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 1.67 (m, 1H), 1.58 (m, 2H), 0.85 (d, J = 6.8 Hz, 6H).

[0071] Example 5:

[0072]

[0073] The preparation steps are the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride is replaced with 4-hydroxybenzenesulfonyl chloride, and other conditions remain unchanged.

[0074] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 9.46 (s, 1H), 7.67 (br s, 1H), 7.53(d, J = 7.5 Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19(t, J = 7.5 Hz, 1H), 7.06 (d, J = 7.5 Hz, 2H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m, 6H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.21-1.96 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0075] Example 6:

[0076]

[0077] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with (S)-3-hydroxytetrahydrofuran, and other conditions remained unchanged.

[0078] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz,1H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m, 6H), 2.92-2.47 (m, 6H), 2.21-1.96 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0079] Example 7:

[0080]

[0081] The preparation steps are the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride is replaced with pyridine-3-sulfonyl chloride, and other conditions remain unchanged.

[0082] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz, 1H), 8.43 (d, J = 7.5 Hz, 1H), 7.59 (dd, J = 7.5, 4.8 Hz, 1H), 7.26 (t, J =7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s,2H), 5.37 (s, 1H), 4.10-3.70 (m, 7H), 2.92-2.47 (m, 6H), 2.19-1.94 (m, 2H),1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0083] Example 8:

[0084]

[0085] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with 3-amino-1-propanol, and other conditions remained unchanged.

[0086] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 9.46 (s, 1H), 7.67 (br s, 1H), 7.53(d, J = 7.5 Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19(t, J = 7.5 Hz, 1H), 7.06 (d, J = 7.5 Hz, 2H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m, 6H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.21-1.96 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0087] Example 9:

[0088]

[0089] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with 1,3-propanediol, and other conditions remained unchanged.

[0090] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz, 1H), 8.43 (d, J = 7.5 Hz, 1H), 7.59 (dd, J = 7.5, 4.8 Hz, 1H), 7.26 (t, J =7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s,1H), 6.02 (br s, 1H), 5.37 (s, 1H), 4.39 (s, 1H), 4.07 (m, 1H), 3.88 (m, 1H), 3.41 (t, J = 7.1 Hz, 2H), 3.38 (t, J = 7.1 Hz, 2H), 2.92-2.47 (m, 6H), 1.67(m, 1H), 1.58 (quint, J = 7.1 Hz, 2H), 0.85 (d, J = 6.8 Hz, 6H).

[0091] Example 10:

[0092]

[0093] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with benzenesulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with (S)-3-hydroxytetrahydrofuran, and other conditions remained unchanged.

[0094] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.88 (d, J = 7.5 Hz, 2H), 7.67 (brs, 1H), 7.64 (t, J = 7.5 Hz, 2H), 7.63 (t, J = 7.5 Hz, 1H), 7.26 (t, J = 7.5Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m, 6H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68(m, 2H), 2.21-1.96 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0095] Example 11:

[0096]

[0097] The preparation steps are the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride is replaced with benzenesulfonyl chloride, and other conditions remain unchanged.

[0098] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.88 (d, J = 7.5 Hz, 2H), 7.64 (t,J = 7.5 Hz, 2H), 7.63 (t, J = 7.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t,J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s, 2H), 4.10-3.70 (m,7H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.19-1.94 (m, 2H),1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0099] Example 12:

[0100]

[0101] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with benzenesulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with 3-amino-1-propanol, and other conditions remained unchanged.

[0102] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 7.88 (d, J = 7.5 Hz, 2H), 7.64 (t,J = 7.5 Hz, 2H), 7.63 (t, J = 7.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t,J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s, 1H), 6.02 (br s, 1H), 5.37 (s, 1H), 4.39 (s, 1H), 4.07 (m, 1H), 3.88 (m, 1H), 3.42-3.36 (m, 6H),3.08 (m, 2H), 2.92-2.68 (m, 2H), 1.67 (m, 1H), 1.58 (quint, J = 7.1 Hz, 2H), 0.85 (d, J = 6.8 Hz, 6H).

[0103] Example 13:

[0104]

[0105] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with benzenesulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with 1,3-propanediol, and other conditions remained unchanged.

[0106] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 7.88 (d, J = 7.5 Hz, 2H), 7.67 (brs, 1H), 7.64 (t, J = 7.5 Hz, 2H), 7.63 (t, J = 7.5 Hz, 1H), 7.26 (t, J = 7.5Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.37 (s, 1H), 4.70 (s, 1H), 4.13 (t, J = 7.1 Hz, 2H), 4.07 (m, 1H), 3.88 (m, 1H), 3.69 (t,J = 7.1 Hz, 2H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 1.88(quint, J = 7.1 Hz, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0107] Example 14:

[0108]

[0109] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with (S)-3-hydroxytetrahydrofuran, and isobutylamine was replaced with cyclopentylmethylamine, while other conditions remained unchanged.

[0110] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz,1H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m, 6H), 2.92-2.47 (m, 6H), 2.21-1.96 (m, 2H), 1.90-1.63 (m, 8H), 1.46 (m, 1H).

[0111] Example 15:

[0112]

[0113] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, and isobutylamine was replaced with cyclopentylmethylamine, while other conditions remained unchanged.

[0114] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz, 1H), 8.43 (d, J = 7.5 Hz, 1H), 7.59 (dd, J = 7.5, 4.8 Hz, 1H), 7.26 (t, J =7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s,2H), 5.37 (s, 1H), 4.10-3.70 (m, 7H), 2.92-2.47 (m, 6H), 2.19-1.94 (m, 2H), 1.90-1.63 (m, 8H), 1.46 (m, 1H).

[0115] Example 16:

[0116]

[0117] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with 3-amino-1-propanol, and isobutylamine was replaced with cyclopentylmethylamine, while other conditions remained unchanged.

[0118] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz, 1H), 8.43 (d, J = 7.5 Hz, 1H), 7.59 (dd, J = 7.5, 4.8 Hz, 1H), 7.26 (t, J =7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s,1H), 6.02 (br s, 1H), 5.37 (s, 1H), 4.39 (s, 1H), 4.07 (m, 1H), 3.88 (m, 1H),3.41 (t, J = 7.1 Hz, 2H), 3.38 (t, J = 7.1 Hz, 2H), 2.92-2.47 (m, 6H), 1.90-1.63 (m, 8H), 1.58 (quint, J = 7.1 Hz, 2H), 1.46 (m, 1H).

[0119] Example 17:

[0120]

[0121] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, (S)-3-aminotetrahydrofuran was replaced with 1,3-propanediol, and isobutylamine was replaced with cyclopentylmethylamine, while other conditions remained unchanged.

[0122] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz,1H), 5.37 (s, 1H), 4.70 (s, 1H), 4.13 (t, J = 7.1 Hz, 2H), 4.07 (m, 1H), 3.88(m, 1H), 3.69 (t, J = 7.1 Hz, 2H), 2.92-2.47 (m, 6H), 1.90-1.63 (m, 8H), 1.88 (quint, J = 7.1 Hz, 2H), 1.46 (m, 1H).

[0123] Example 18:

[0124]

[0125] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, isobutylamine was replaced with 2-furanmethylamine, and (S)-3-aminotetrahydrofuran was replaced with (S)-3-hydroxytetrahydrofuran, while other conditions remained unchanged.

[0126] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.55 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J =7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.39 (dd, J = 7.5, 7.5 Hz, 1H), 6.29(d, J = 7.5 Hz, 1H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.66 (m, 8H), 2.92-2.55(m, 4H), 2.21-1.96 (m, 2H).

[0127] Example 19:

[0128]

[0129] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, and isobutylamine was replaced with 2-furanmethylamine, while other conditions remained unchanged.

[0130] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz, 1H), 8.43 (d, J = 7.5 Hz, 1H), 7.59 (dd, J = 7.5, 4.8 Hz, 1H), 7.55 (dd, J =7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19(t, J = 7.5 Hz, 1H), 6.46 (br s, 2H), 6.39 (dd, J = 7.5, 7.5 Hz, 1H), 6.29(d, J = 7.5 Hz, 1H), 5.37 (s, 1H), 4.10-3.66 (m, 9H), 2.92-2.55 (m, 4H), 2.19-1.94 (m, 2H).

[0131] Example 20:

[0132]

[0133] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, isobutylamine was replaced with 2-furanmethylamine, and (S)-3-aminotetrahydrofuran was replaced with (3-amino-1-propanol), while other conditions remained unchanged.

[0134] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz, 1H), 8.43 (d, J = 7.5 Hz, 1H), 7.59 (dd, J = 7.5, 4.8 Hz, 1H), 7.55 (dd, J =7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19(t, J = 7.5 Hz, 1H), 6.46 (br s, 1H), 6.39 (dd, J = 7.5, 7.5 Hz, 1H), 6.29(d, J = 7.5 Hz, 1H), 6.02 (br s, 1H), 5.37 (s, 1H), 4.39 (s, 1H), 4.07 (m,1H), 3.88 (m, 1H), 3.66 (s, 2H), 3.41 (t, J = 7.1 Hz, 2H), 3.38 (t, J = 7.1Hz, 2H), 2.92-2.55 (m, 4H), 1.58 (quint, J = 7.1 Hz, 2H).

[0135] Example 21:

[0136]

[0137] The preparation steps were the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride was replaced with pyridine-3-sulfonyl chloride, isobutylamine was replaced with 2-furanmethylamine, and (S)-3-aminotetrahydrofuran was replaced with 3-amino-1-propanol, while other conditions remained unchanged.

[0138] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.55 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J =7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.39 (dd, J = 7.5, 7.5 Hz, 1H), 6.29(d, J = 7.5 Hz, 1H), 5.37 (s, 1H), 4.70 (s, 1H), 4.13 (t, J = 7.1 Hz, 2H), 4.07 (m, 1H), 3.88 (m, 1H), 3.69 (t, J = 7.1 Hz, 2H), 3.66 (s, 2H), 2.92-2.55(m, 4H), 1.88 (quint, J = 7.1 Hz, 2H).

[0139] Example 22:

[0140]

[0141] The preparation steps were the same as in Example 1, except that (S)-3-aminotetrahydrofuran was replaced with 1,3-propanediol, and other conditions remained unchanged.

[0142] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 7.67 (br s, 1H), 7.55 (d, J = 7.5Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5Hz, 1H), 6.58 (d, J = 7.5 Hz, 2H), 6.06 (br s, 2H), 5.37 (s, 1H), 4.70 (s,1H), 4.13 (t, J = 7.1 Hz, 2H), 4.07 (m, 1H), 3.88 (m, 1H), 3.69 (t, J = 7.1Hz, 2H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 1.88 (quint, J =7.1 Hz, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0143] Example 23:

[0144]

[0145] The synthesis route is shown below:

[0146]

[0147] Step 1: Preparation steps are as described in Example 1.

[0148] Step 2: The preparation steps are the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride is replaced with benzenesulfonyl chloride, and other conditions remain unchanged.

[0149] Step 3: Synthesis of (2R,3S)-4-phenyl-1-((isobutyl((phenyl)sulfonyl)amino)-3-aminobut-2-ol)

[0150] (2S,3R)-tert-butyl((1-hydroxy-3-((isobutyl)((phenyl)sulfonyl)amino)-4-phenylbut-2-yl)carbamate (2 g, 1 equiv) was placed in a round-bottom flask, and 10 mL of trifluoroacetic acid was added dropwise with 30–50 mL of dichloromethane as solvent. The mixture was stirred at room temperature for 2 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was completed, the solvent was evaporated, and the target compound (1.42 g, white solid, yield 90.4%) was obtained by extraction and column chromatography.

[0151] Step 4: Synthesis of (2S)-N-(benzenesulfonyl)-4-methyl-2-({(2S,3R)-2-hydroxy-3-[({[2-((3aS,6aR)-hexahydrofurano[3,2-b]furan-3-yl)oxy]-2-oxoacetyl}amino)methyl]propionyl}amino)pentanamide

[0152] (2R,3S)-4-phenyl-1-((isobutyl((phenyl)sulfonyl)amino)-3-aminobut-2-ol (1 g, 1 equiv), 2,5-dioxopyrrolidine-1-carboxylic acid (3R,3aS,6S,6aR)-hexahydrofurano[3,2-b]furan-3-ylmethyl ester (1.14 g, 1.5 equiv), and triethylamine (0.81 g, 3 equiv) were placed in a round-bottom flask with 20-30 M dichloromethane as solvent. The mixture was stirred at room temperature for 12 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was completed, the solvent was evaporated, and the product was extracted and subjected to column chromatography to obtain the target compound (1.18 g, white solid, yield 83.6%).

[0153] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 7.88 (d, J = 7.5 Hz, 2H), 7.67 (brs, 1H), 7.64 (t, J = 7.5 Hz, 2H), 7.63 (t, J = 7.5 Hz, 1H), 7.26 (t, J = 7.5Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.45 (d, J = 9.3Hz, 1H), 5.37 (s, 1H), 4.92 (m, 1H), 4.12-3.70 (m, 6H), 3.42-3.16 (m, 2H),3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.54 (m, 1H), 1.92-1.67 (m, 3H), 0.85 (d, J= 6.8 Hz, 6H).

[0154] Example 24:

[0155]

[0156] The preparation steps are the same as in Example 4, except that p-benzenesulfonyl chloride is replaced with pyridine-3-sulfonyl chloride, and other conditions remain unchanged.

[0157] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz,1H), 5.45 (d, J = 9.3 Hz, 1H), 5.37 (s, 1H), 4.92 (m, 1H), 4.12-3.70 (m, 6H), 2.92-2.55 (m, 6H), 2.54 (m, 1H), 1.92-1.66 (m, 3H), 0.85 (d, J = 6.8 Hz, 6H).

[0158] Example 25:

[0159]

[0160] The preparation steps are the same as in Example 4, except that p-benzenesulfonyl chloride is replaced with pyridine-3-sulfonyl chloride, isobutylamine is replaced with cyclopentylmethylamine, and other conditions remain unchanged.

[0161] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz,1H), 5.45 (d, J = 9.3 Hz, 1H), 5.37 (s, 1H), 4.92 (m, 1H), 4.12-3.70 (m, 5H), 2.92-2.47 (m, 6H), 2.54 (m, 1H), 1.92-1.63 (m, 10H), 1.46 (m, 1H).

[0162] Example 26:

[0163]

[0164] The preparation steps are the same as in Example 4, except that p-benzenesulfonyl chloride is replaced with pyridine-3-sulfonyl chloride, isobutylamine is replaced with 2-furanmethylamine, and other conditions remain unchanged.

[0165] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.91 (s, 1H), 8.85 (d, J = 7.5 Hz,1H), 8.43 (d, J = 7.5 Hz, 1H), 7.67 (br s, 1H), 7.59 (dd, J = 7.5, 4.8 Hz,1H), 7.55 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J =7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.39 (dd, J = 7.5, 7.5 Hz, 1H), 6.29(d, J = 7.5 Hz, 1H), 5.45 (d, J = 9.3 Hz, 1H), 5.37 (s, 1H), 4.92 (m, 1H), 4.12-3.66 (m, 8H), 2.92-2.55 (m, 4H), 2.54 (m, 1H), 1.92-1.66 (m, 2H).

[0166] Example 27:

[0167]

[0168] The synthesis route is shown below:

[0169]

[0170] Step 1: The preparation steps are the same as in Example 1.

[0171] Step 2: The preparation steps are the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride is replaced with 4-bromobenzenesulfonyl chloride, and other conditions remain unchanged.

[0172] Step 3: The preparation steps are the same as in Example 1.

[0173] Step 4: Synthesis of N-((2R)-2-acetoxy-3-benzyl-4-((tert-butoxycarbonyl)amino)butyl)-N-isobutyl-4-(4-methylpiperazin-1-yl)benzenesulfonamide

[0174] (2S,3R)-1-(N-isobutyl-4-bromobenzenesulfonylamino)-3-acetoxy-4-phenylbut-2-ylcarbamate tert-butyl ester (1 g, 1 equiv), N-methylpiperazine (0.3 g, 1.8 equiv), tetrabutylammonium bromide (0.05 g, 0.1 equiv), and potassium tert-butoxy (0.42 g, 2 equiv) were placed in a round-bottom flask and stirred at 120 °C for 12 h under nitrogen protection with 20–30 M dimethyl sulfoxide as solvent. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solvent was evaporated, and the product was extracted and subjected to column chromatography to obtain the target compound (0.6 g, white solid, yield 61.2%).

[0175] Step 5: The preparation steps are the same as in Example 1.

[0176] Step 6: The preparation steps are the same as in Example 1.

[0177] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.52 (d, J = 7.5 Hz, 2H), 7.26 (t,J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.98 (d,J = 7.5 Hz, 2H), 6.46 (br s, 2H), 5.37 (s, 1H), 4.10-3.70 (m, 6H), 3.44 (t, J= 7.1 Hz, 4H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.35 (t, J= 7.1 Hz, 4H), 2.21 (s, 3H), 2.19-1.94 (m, 2H), 1.67 (m, 1H), 0.85 (d, J =6.8 Hz, 6H).

[0178] Example 28:

[0179]

[0180] The preparation steps were the same as in Example 27, except that (S)-3-aminotetrahydrofuran was replaced with (S)-hydroxytetrahydrofuran, and 4-bromobenzenesulfonyl chloride was replaced with 5-bromo-2-pyridinesulfonyl chloride, while other conditions remained unchanged.

[0181] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.07 (d, J = 7.5 Hz, 1H), 7.90 (brs, 1H), 7.67 (br s, 1H), 7.35 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz,2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.37 (s, 1H), 5.02(m, 1H), 4.12-3.70 (m, 6H), 3.15 (t, J = 7.1 Hz, 4H), 2.92-2.55 (m, 6H), 2.35(t, J = 7.1 Hz, 4H), 2.21 (s, 3H, overlaps with m, 2H from THF), 1.67 (m,1H), 0.85 (d, J = 6.8 Hz, 6H).

[0182] Example 29:

[0183]

[0184] The preparation steps are the same as in Example 27, except that (S)-3-aminotetrahydrofuran is replaced with (S)-hydroxytetrahydrofuran, and other conditions remain unchanged.

[0185] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.67 (br s, 1H), 7.52 (d, J = 7.5Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5Hz, 1H), 6.98 (d, J = 7.5 Hz, 2H), 5.37 (s, 1H), 5.02 (m, 1H), 4.12-3.70 (m,6H), 3.44 (t, J = 7.1 Hz, 4H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.35 (t, J = 7.1 Hz, 4H), 2.21 (s, 3H, overlaps with m, 2H), 1.67 (m,1H), 0.85 (d, J = 6.8 Hz, 6H).

[0186] Example 30:

[0187]

[0188] The preparation steps were the same as in Example 27, except that (S)-3-aminotetrahydrofuran was replaced with 3-amino-1-propanol, 4-bromobenzenesulfonyl chloride was replaced with 5-bromo-2-pyridinesulfonyl chloride, and other conditions remained unchanged.

[0189] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.07 (d, J = 7.5 Hz, 1H), 7.90 (brs, 1H), 7.35 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J= 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s, 1H), 6.02 (br s, 1H), 5.37 (s, 1H), 4.39 (s, 1H), 4.07 (m, 1H), 3.88 (m, 1H), 3.41 (t, J = 7.1 Hz,2H), 3.38 (t, J = 7.1 Hz, 2H), 3.15 (t, J = 7.1 Hz, 4H), 2.92-2.47 (m, 6H), 2.35 (t, J = 7.1 Hz, 4H), 2.21 (s, 3H), 1.67 (m, 1H), 1.58 (quint, J = 7.1Hz, 2H), 0.85 (d, J = 6.8 Hz, 6H).

[0190] Example 31:

[0191]

[0192] The preparation steps were the same as in Example 27, except that p-(S)-3-aminotetrahydrofuran was replaced with 1,3-propanediol, 4-bromobenzenesulfonyl chloride was replaced with 5-bromo-2-pyridinesulfonyl chloride, and other conditions remained unchanged.

[0193] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.07 (d, J = 7.5 Hz, 1H), 7.90 (brs, 1H), 7.67 (br s, 1H), 7.35 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz,2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.37 (s, 1H), 4.70(s, 1H), 4.13 (t, J = 7.1 Hz, 2H), 4.07 (m, 1H), 3.88 (m, 1H), 3.69 (t, J =7.1 Hz, 2H), 3.15 (t, J = 7.1 Hz, 4H), 2.92-2.47 (m, 6H), 2.35 (t, J = 7.1Hz, 4H), 2.21 (s, 3H), 1.88 (quint, J = 7.1 Hz, 2H), 1.67 (m, 1H), 0.85 (d, J= 6.8 Hz, 6H).

[0194] Example 32:

[0195]

[0196] The preparation steps were the same as in Example 27, except that (S)-3-aminotetrahydrofuran was replaced with 3-amino-1-propanol, and other conditions remained unchanged.

[0197] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.07 (d, J = 7.5 Hz, 1H), 7.90 (brs, 1H), 7.67 (br s, 1H), 7.35 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz,2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.37 (s, 1H), 4.70(s, 1H), 4.13 (t, J = 7.1 Hz, 2H), 4.07 (m, 1H), 3.88 (m, 1H), 3.69 (t, J =7.1 Hz, 2H), 3.15 (t, J = 7.1 Hz, 4H), 2.92-2.47 (m, 6H), 2.35 (t, J = 7.1Hz, 4H), 2.21 (s, 3H), 1.88 (quint, J = 7.1 Hz, 2H), 1.67 (m, 1H), 0.85 (d, J= 6.8 Hz, 6H).

[0198] Example 33:

[0199]

[0200] The preparation steps are the same as in Example 27, except that 4-bromobenzenesulfonyl chloride is replaced with 5-bromo-2-pyridinesulfonyl chloride, and other conditions remain unchanged.

[0201] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 8.07 (d, J = 7.5 Hz, 1H), 7.90 (brs, 1H), 7.35 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J= 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.46 (br s, 2H), 4.10-3.70 (m, 7H), 3.15 (t, J = 7.1 Hz, 4H), 2.92-2.47 (m, 6H), 2.35 (t, J = 7.1 Hz, 4H), 2.21(s, 3H), 2.19-1.94 (m, 2H), 1.67 (m, 1H), 0.85 (d, J = 6.8 Hz, 6H).

[0202] Example 34:

[0203]

[0204] The preparation steps were the same as in Example 27, except that p-(S)-3-aminotetrahydrofuran was replaced with 1,3-propanediol, while other conditions remained unchanged.

[0205] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.67 (br s, 1H), 7.52 (d, J = 7.5Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5Hz, 1H), 6.98 (d, J = 7.5 Hz, 2H), 5.37 (s, 1H), 4.70 (s, 1H), 4.13 (t, J =7.1 Hz, 2H), 4.07 (m, 1H), 3.88 (m, 1H), 3.69 (t, J = 7.1 Hz, 2H), 3.44 (t, J= 7.1 Hz, 4H), 3.42-3.16 (m, 2H), 3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.35 (t, J= 7.1 Hz, 4H), 2.21 (s, 3H), 1.88 (quint, J = 7.1 Hz, 2H), 1.67 (m, 1H), 0.85(d, J = 6.8 Hz, 6H).

[0206] Example 35:

[0207]

[0208]

[0209] Step 1: The preparation steps are the same as in Example 1.

[0210] Step 2: The preparation steps are the same as in Example 1, except that 4-nitrobenzenesulfonyl chloride is replaced with 4-bromobenzenesulfonyl chloride, and other conditions remain unchanged.

[0211] Step 3: The preparation steps are the same as in Example 1.

[0212] Step 4: The preparation steps are the same as in Example 27.

[0213] Step 5: The preparation steps are the same as in Example 23.

[0214] Step 6: The preparation steps are the same as in Example 23.

[0215] Step 6: The preparation steps are the same as in Example 27.

[0216] 1 H NMR (300 MHz, DMSO-d6) δ (ppm) 7.67 (br s, 1H), 7.52 (d, J = 7.5Hz, 2H), 7.26 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5Hz, 1H), 6.98 (d, J = 7.5 Hz, 2H), 5.45 (d, J = 9.3 Hz, 1H), 5.37 (s, 1H), 4.92 (m, 1H), 4.12-3.70 (m, 6H), 3.44 (t, J = 7.1 Hz, 4H), 3.42-3.16 (m, 2H),3.08 (m, 2H), 2.92-2.68 (m, 2H), 2.54 (m, 1H), 2.35 (t, J = 7.1 Hz, 4H), 2.21(s, 3H), 1.92-1.66 (m, 3H), 0.85 (d, J = 6.8 Hz, 6H).

[0217] Example 36:

[0218]

[0219] The preparation steps are the same as in Example 35, except that 4-bromobenzenesulfonyl chloride is replaced with 5-bromo-2-pyridinesulfonyl chloride, and other conditions remain unchanged.

[0220] 1H NMR (300 MHz, DMSO-d6) δ (ppm) 8.07 (d, J = 7.5 Hz, 1H), 7.90 (brs, 1H), 7.67 (br s, 1H), 7.35 (dd, J = 7.5, 1.5 Hz, 1H), 7.26 (t, J = 7.5 Hz,2H), 7.24 (t, J = 7.5 Hz, 2H), 7.19 (t, J = 7.5 Hz, 1H), 5.45 (d, J = 9.3 Hz,1H), 5.37 (s, 1H), 4.92 (m, 1H), 4.12-3.70 (m, 6H), 3.15 (t, J = 7.1 Hz, 4H), 2.92-2.47 (m, 6H), 2.54 (m, 1H), 2.35 (t, J = 7.1 Hz, 4H), 2.21 (s, 3H), 1.92-1.66 (m, 3H), 0.85 (d, J = 6.8 Hz, 6H).

[0221] The inhibitory activity of the compounds obtained in the above examples against pepsin was determined by the following method:

[0222] Dilute pepsin to a suitable activity concentration using sodium acetate-hydrochloric acid buffer (0.06 M, pH=2.0) and set aside. Dissolve approximately 5 mg of the test compound in DMSO solution and dilute with sodium acetate-hydrochloric acid buffer to the desired concentration gradient (ensuring the DMSO content in the prepared solutions does not exceed 1%). Add 200 μL of substrate solution (2% denatured hemoglobin, dissolved in pH 2.0 buffer), 80 μL of diluted pepsin solution, and 20 μL of compounds at different concentration gradients to test tubes or 96-well plates sequentially. For the blank control group, add 20 μL of sodium acetate-hydrochloric acid buffer (or add stop solution first to eliminate background). Incubate at 37°C for 10-20 mins (the exact time needs to be determined in a preliminary experiment) to allow the test compound to bind to pepsin, thereby effectively inhibiting the hydrolytic reaction between pepsin and the substrate hemoglobin. After the incubation period, 400 μL of stop solution (5% trichloroacetic acid, TCA) was added to each well to precipitate unhydrolyzed large protein molecules, bringing the total volume of the reaction system to 700 μL (according to this ratio). The reaction was terminated and allowed to stand at room temperature for 10 min. The supernatant was then centrifuged and immediately analyzed using a UV-Vis spectrophotometer or microplate reader (absorbance values ​​were read at 280 nm). This allowed for the calculation of the amount of soluble peptides produced by pepsin hydrolysis of hemoglobin. GraphPad Prism 9.5 software was used to plot the inhibition rate data at different concentrations, thereby calculating the IC50 value of the compound.

[0223] The water solubility of the compounds obtained in the above examples was determined by the following method:

[0224] Accurately weigh the analyte (approximately 5-10 mg) and place it in a clean, dry glass tube or a 96-well deep-well plate. Add an appropriate amount of general-purpose buffer (such as pH 7.4 phosphate-buffered saline (PBS), or sodium acetate-hydrochloric acid buffer (pH 2.0 / pure water) selected according to the target physiological environment) as the solvent medium. First, add a small amount of dimethyl sulfoxide (DMSO) to the sample to aid dissolution (controlling the initial DMSO volume fraction to no more than 5%), and vortex to completely dissolve the compound and form a high-concentration stock solution. Subsequently, under vigorous vortexing, add the above buffer dropwise for gradient dilution until visible turbidity or precipitation is observed in the solution, recording this as a supersaturated state; alternatively, directly prepare a series of concentration gradient solutions with higher than the expected solubility (e.g., 1 μM to 10 mM), place them in a constant-temperature shaker, and incubate at 25°C (or 37°C) at a speed of 200-300 rpm for 24 hours to ensure dissolution equilibrium is reached.

[0225] After equilibration, allow the sample tubes to stand at room temperature for 10-30 minutes to allow undissolved large particles to settle, or filter directly using a 0.45 μm or 0.22 μm microporous membrane (the material should be selected according to the compound properties, either PTFE or PVDF). Discard the initial filtrate and collect the clear filtrate as the sample to be tested. If UV-Vis spectrophotometry is used for detection, immediately take an appropriate amount of filtrate and perform absorbance measurement using a UV-Vis spectrophotometer or microplate reader (select the maximum absorption wavelength of the compound, usually 254 nm, 280 nm, or determined based on a full wavelength scan). Simultaneously, use a blank buffer (containing an equal proportion of DMSO) without the compound as a reference control to subtract the background absorbance. Calculate the actual concentration of the compound in the filtrate using a pre-plotted standard curve, which is the equilibrium solubility under these conditions. If high-performance liquid chromatography (HPLC) is used for detection (recommended for patent data to confirm purity), inject an appropriate amount of filtrate, separate it through a chromatographic column, and detect the peak area at a specific wavelength. Calculate the concentration using an external standard curve.

[0226] The results of the bioactivity experiments and water solubility of compounds 1-36 and positive controls against MAOs are shown in the table below.

[0227]

[0228] a, b Each IC 50 It is the average of three experiments, IC 50 The standard error is generally less than 10%.

[0229] NA d When the inhibitory concentration is greater than 100 μM, it indicates no activity.

Claims

1. An aminobenzenesulfonamide derivative, characterized in that... Its general structural formula is as follows: ； In the general formula: X is O, S, or N; R1 is selected from , , , , m can be 1, 2, 3 or 4; R2 is selected from , , , , , , , , , , , ; R3 is selected from hydrogen, deuterium, halogen, methyl, hydroxyl, methoxy, ethoxy, hydroxymethyl, amino, nitro. , , , , , , , , , , , , , , , m can be 1, 2, 3, or 4. Ring A is selected from one of the following: benzene ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring, 1,3,5-triazine ring, 1,2,4-triazine ring, and 1,2,3-triazine ring.

2. The aminobenzenesulfonamide derivative according to claim 1, characterized in that... Selected from compounds with the following structures: ； ； ； ； ； 。 3. The use of the aminobenzenesulfonamide derivative of claim 1 in the preparation of pharmaceutical formulations.

4. The application according to claim 3, characterized in that: The pharmaceutical preparation is a pharmaceutical preparation for the prevention and / or treatment of pepsin-related diseases.

5. The application according to claim 4, characterized in that: The pepsin-related diseases mentioned include laryngopharyngeal reflux disease.

6. The application according to claim 5, characterized in that: The laryngopharyngeal reflux disease includes one or more of the following: reflux laryngitis, laryngopharyngeal reflux disease, chronic cough caused by reflux, vocal cord nodules, vocal cord polyps, and foreign body sensation in the throat.

7. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the aminobenzenesulfonamide derivative or a pharmaceutically acceptable salt, cocrystal, or solvate thereof.

8. The pharmaceutical composition according to claim 7, characterized in that: The pharmaceutical composition is formulated as a spray.

9. The pharmaceutical composition according to claim 7, characterized in that: The pharmaceutical composition further includes pharmaceutically acceptable excipients and / or carriers.

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