Benzene sulfonamide aryl amide derivatives and uses thereof

Abstract

The application discloses a benzene sulfonamide aryl amide derivative and application thereof. The series of compounds are verified by pharmacological tests to have no direct antibacterial activity, can be used as a bacterial quorum sensing inhibitor to block a bacterial quorum sensing path, inhibit biofilm formation and secretion of a virulence factor, and reduce pathogenicity of bacteria, and can be used as an antibacterial sensitizer to improve a bactericidal effect of an antibiotic on multi-drug resistant bacteria, thereby providing a novel lead compound and technical thought for combined anti-infection treatment of the multi-drug resistant bacteria in clinic.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of drug synthesis, and particularly relates to a benzene sulfonamide aryl amide derivative and application thereof. BACKGROUND

[0002] The long-term and extensive use of antibiotics has greatly accelerated the occurrence and spread of bacterial drug resistance, making multi-drug resistant bacterial infections a major problem threatening global public health security. The World Health Organization (WHO) has listed antibiotic resistance as one of the top ten global public health threats. Traditional anti-infective drugs are mostly bactericidal or bacteriostatic antibiotics, which mainly interfere with the basic life processes necessary for bacterial survival and proliferation, such as bacterial cell wall synthesis, protein expression, DNA replication and transcription. Such mechanisms of action will exert strong selective pressure on bacteria, rapidly selecting and enriching drug-resistant mutant strains in the drug environment, and thus inducing widespread and severe drug resistance. Even though new antibiotics are continuously developed, their clinical effective use period is generally significantly shortened due to the rapid spread of drug-resistant strains, so it is urgent to develop new anti-infective strategies that are different from traditional bactericidal mechanisms.

[0003] Anti-virulence strategies have emerged as the core is not to directly kill or inhibit bacterial growth, but to target the virulence-related pathways of pathogenic bacteria to weaken the pathogenic ability of pathogens, thereby reducing infection symptoms and creating favorable conditions for host immune clearance. This strategy can not only reduce the generation of drug resistance, but also effectively curb the spread and diffusion of drug-resistant pathogenic bacteria, so it is considered as a promising research direction in the post-antibiotic era.

[0004] Quorum sensing (QS) is a core communication mechanism by which bacteria monitor population density by secreting and sensing autoinducer signal molecules, and coordinate group behavior accordingly. The quorum sensing system widely regulates various virulence-related phenotypes, including biofilm formation, virulence factor secretion, bacterial swarm motility, exopolysaccharide formation, and cell differentiation. Therefore, small molecule inhibitors targeting the quorum sensing system, by interfering with the synthesis, release, perception or downstream signal transduction of signal molecules, are expected to effectively "disarm" the virulence of pathogenic bacteria without directly killing bacteria, thereby reducing the severity of infection and reducing the selection pressure of drug resistance, providing a promising path for the development of new anti-infective drugs.

[0005] Currently, some natural products and synthetic small molecules have been shown to possess quorum sensing inhibition (QSI) activity, such as bromofuranones, flavonoids, and acyl homoserine lactone (AHL) structural analogs derived from marine fungi. However, no QSI drugs have yet been approved for clinical treatment of bacterial infections, especially for refractory pathogens such as Klebsiella pneumoniae and Pseudomonas aeruginosa, which are prevalent, highly virulent, and highly drug-resistant. There is a severe lack of compounds with novel structures, high safety profiles, and the ability to combine quorum sensing inhibition with antibiotic sensitization. Summary of the Invention

[0006] To address the problems of the prior art, the present invention aims to provide a novel class of benzenesulfonamide-based aryl amide derivatives and their applications. These derivatives exhibit no significant direct antibacterial activity but possess the dual application value of quorum sensing inhibition and broad-spectrum antibacterial sensitization.

[0007] This invention is achieved through the following technical solution:

[0008] In a first aspect, the present invention provides a benzenesulfonamide-based arylamide derivative or a pharmaceutically acceptable salt or hydrate thereof, said derivative having a structure as shown in general formula I or II:

[0009]

[0010] Wherein, R is selected from C1-C8 straight-chain or branched alkyl, C3-C6 cycloalkyl, phenyl with different substituents, aryl with different substituents, C3-C6 arylcyclomethyl, and adamantyl.

[0011] R1 is selected from hydrogen, C1-C8 straight-chain or branched alkyl groups, halogens, and C1-C8 straight-chain or branched alkoxy groups.

[0012] R2 and R3 may be the same or different, and are independently selected from hydrogen, C1-C8 straight-chain or branched alkyl groups.

[0013] In a preferred embodiment of the present invention, R is selected from C3-C6 straight-chain or branched alkyl, C3-C6 cycloalkyl, phenyl with alkyl or trifluoromethyl substituents, aryl with alkoxycarbonyl substituents, C3-C6 arylcyclomethyl, and adamantyl.

[0014] R1 is selected from hydrogen, C1-C8 straight-chain alkyl, halogen, and C1-C8 straight-chain alkoxy.

[0015] R2 and R3 may be the same or different, and are independently selected from straight-chain alkyl groups of C1-C8.

[0016] In a preferred embodiment of the present invention, R1 is selected from methoxy, fluorine, and methyl; R2 is methyl; and R3 is methyl.

[0017] In a further preferred embodiment of the present invention, R is selected from isopropyl, cyclohexyl, 4-ethylphenyl, 4-trifluoromethylphenyl, 3-trifluoromethylphenyl, 2-methylphenyl, 2-ethylphenyl, 4-ethoxycarbonylphenyl, 4-diethylaminophenyl, 5-ethoxycarbonyl-2-thiazolyl, 4-methoxycarbonyl-2-thiazolyl, 2-furanmethyl, and adamantyl.

[0018] R1 is selected from methoxy, fluorine, and methyl; R2 is methyl; R3 is methyl.

[0019] In a preferred embodiment of the present invention, the benzenesulfonamide-arylamide derivative is selected from any of the following compounds:

[0020]

[0021]

[0022]

[0023]

[0024]

[0025]

[0026]

[0027]

[0028]

[0029]

[0030]

[0031]

[0032]

[0033]

[0034]

[0035]

[0036] .

[0037] Secondly, the benzenesulfonamide-aryl amide derivatives of general formula I or II of the present invention are prepared according to the following two reaction routes, specifically:

[0038] Route 1: Benzenesulfonyl chloride (A) with different substituents is sulfonated with ethyl p-aminobenzoate (B) to obtain intermediate M1 and intermediate M2. Intermediate M1 is hydrolyzed to obtain intermediate M2. Intermediate M2 undergoes a condensation reaction with different substituted amino groups to obtain the compound shown in Formula I.

[0039]

[0040] Route 2: The intermediate M3 is obtained by sulfonation of p-methoxybenzenesulfonyl chloride (C) and ethyl 2-aminothiazolyl carboxylate (D). Intermediate M3 is hydrolyzed to obtain intermediate M4. Intermediate M4 undergoes condensation reaction with different substituted amino groups to obtain the compound shown in Formula II.

[0041]

[0042] Wherein, R is selected from C1-C8 straight-chain or branched alkyl, C3-C6 cycloalkyl, phenyl with different substituents, aryl with different substituents, C3-C6 arylcyclomethyl, and adamantyl.

[0043] R1 is selected from hydrogen, C1-C8 straight-chain or branched alkyl groups, halogens, and C1-C8 straight-chain or branched alkoxy groups.

[0044] R2 and R3 may be the same or different, and are independently selected from hydrogen, C1-C8 straight-chain or branched alkyl groups.

[0045] Thirdly, the present invention provides a pharmaceutical composition comprising the above-mentioned benzenesulfonamide aryl amide derivatives or their pharmaceutically acceptable salts or hydrates, and a pharmaceutically acceptable carrier or excipient.

[0046] In a preferred embodiment of the present invention, the pharmaceutical composition further includes at least one antibacterial active ingredient, wherein the antibacterial active ingredient is kanamycin, streptomycin, azithromycin, chloramphenicol, ciprofloxacin, clarithromycin, tobramycin, norfloxacin, or levofloxacin.

[0047] The pharmaceutically acceptable salts described in this invention include one or more of inorganic and organic salts. The inorganic salts refer to salts formed with inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, and hydrobromic acid. The organic salts refer to salts formed with organic acids, such as acetic acid, citric acid, maleic acid, oxalic acid, succinic acid, lactic acid, malic acid, propionic acid, tartaric acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid.

[0048] The pharmaceutically acceptable carriers or excipients described in this invention refer to excipients that are pharmacologically and / or physiologically compatible with the subject and the active ingredient. In some embodiments, the pharmaceutically acceptable excipients include at least one of solvents, disintegrants, flavoring agents, preservatives, colorants, binders, fillers, stabilizers, antioxidants, and lubricants.

[0049] The pharmaceutical compositions of the present invention can be prepared into any pharmaceutically acceptable dosage form by methods known in the art. In some embodiments, the dosage form is a tablet, pill, capsule, suspension, emulsion, powder, lyophilized powder for injection, aerosol, injection, inclusion compound, implant, patch, or liniment.

[0050] Fourthly, the present invention also provides the use of the above-mentioned benzenesulfonamide arylamide derivatives or their pharmaceutically acceptable salts or hydrates or the above-mentioned pharmaceutical compositions in the preparation of bacterial quorum sensing inhibitors or antibacterial sensitizers.

[0051] In some embodiments, the bacteria are Gram-positive or Gram-negative bacteria, preferably Gram-negative bacteria; the Gram-negative bacteria include, but are not limited to, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, Salmonella, Shigella, Salmonella typhi, and Proteus, preferably Klebsiella pneumoniae.

[0052] Fifthly, the present invention also provides the use of the above-described benzenesulfonamide arylamide derivatives or their pharmaceutically acceptable salts or hydrates, or the above-described pharmaceutical compositions, in the preparation of medicaments for the prevention and / or treatment of diseases caused by bacterial infections.

[0053] In some embodiments, the diseases caused by bacterial infection described in this invention include, but are not limited to, pneumonia, urinary tract infection, cystitis, diarrhea, sepsis, cholecystitis, endocarditis, and gastroenteritis.

[0054] The dosage of the drug of this invention can be varied according to the route of administration, the patient's age, weight, the type and severity of the disease being treated, etc.

[0055] Compared with the prior art, the present invention has the following beneficial effects:

[0056] This invention provides a series of novel benzenesulfonamide-based arylamide derivatives. Pharmacological experiments have verified that this series of compounds have no direct antibacterial activity. They can act as bacterial quorum sensing inhibitors, blocking bacterial quorum sensing pathways, inhibiting biofilm formation and virulence factor secretion, and reducing bacterial pathogenicity. They can also act as antibacterial sensitizers, enhancing the bactericidal effect of antibiotics against multidrug-resistant bacteria. This provides a novel lead compound and technical approach for the combined anti-infective treatment of multidrug-resistant bacteria in clinical practice. Attached Figure Description

[0057] Figure 1 The figure shows the results of investigating the inhibitory activity of compounds 1-34 on AI-2 generation of clinical multidrug-resistant Klebsiella pneumoniae strain KP31 in the embodiments of the present invention.

[0058] Figure 2 The figure shows the results of investigating the inhibitory activity of compounds 1-34 on biofilm formation of Klebsiella pneumoniae clinical multidrug-resistant strain KP31 in the embodiments of the present invention;

[0059] Figure 3 The figure shows the results of investigating the inhibitory activity of compounds 1, 6, 7, 8, 9, 10, 11, and 13 against the cluster motility of clinically multidrug-resistant Klebsiella pneumoniae strain KP31 in the embodiments of the present invention.

[0060] Figure 4 The figure shows the results of investigating the inhibitory activity of compounds 1, 6, 8, 11, and 13 against the capsular polysaccharide of Klebsiella pneumoniae clinically multidrug-resistant strain KP31 in the embodiments of the present invention.

[0061] Figure 5 The embodiments of the present invention demonstrate the synergistic effect of compound 1 with the antibacterial agents kanamycin, streptomycin, azithromycin, and chloramphenicol against the clinically multidrug-resistant Klebsiella pneumoniae strain KP31. Detailed Implementation

[0062] The present invention is further illustrated below by way of embodiments, but the implementation of the present invention is not limited to the following embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention should be considered equivalent substitutions and are included within the protection scope of the present invention. Experimental methods in the following embodiments that do not specify specific conditions are performed according to conventional methods and conditions, or according to the product instructions.

[0063] The initial raw materials and reaction reagents used in the specific embodiments of this invention are all commercially available products.

[0064] Example 1: Synthesis of N-(4-ethylphenyl)-4-((4-methoxyphenyl)sulfonamide)benzamide (Compound 1)

[0065]

[0066] (1) Synthetic intermediate 1-M1

[0067] 1 mmol of ethyl p-aminobenzoate was dissolved in dichloromethane, and 7.5 mmol of pyridine was added. The mixture was stirred and the reaction was carried out. Then, 1.2 mmol of p-methoxybenzenesulfonyl chloride dissolved in dichloromethane was slowly added dropwise to the reaction solution. The reaction was carried out at room temperature and monitored by TLC. After the reaction was completed, the reaction solution was added to a saturated sodium bicarbonate solution and extracted three times with dichloromethane. The dichloromethane layers were combined and washed with hydrochloric acid solution and water, respectively. The organic layer was concentrated to obtain intermediate 1-M1.

[0068] (2) Synthetic intermediate 1-M2

[0069] 1-M1 was dissolved in methanol, and 4 times the volume of lithium hydroxide aqueous solution was added. The reaction was stirred at 50°C. The hydrolysis reaction was monitored by TLC until it was complete. The methanol was removed by concentration, and the pH was adjusted to acidic with dilute hydrochloric acid. A large amount of precipitate was formed. The precipitate was filtered and washed with water several times to obtain intermediate 1-M2.

[0070] (3) Synthesis of N-(4-ethylphenyl)-4-((4-methoxyphenyl)sulfonamide)benzamide (compound 1)

[0071]

[0072] 1-M2 was completely dissolved in dry DMF, and 1.5 equivalents of HATU and 2 equivalents of triethylamine were added. The mixture was magnetically stirred and activated for 30 min. Then, 1.2 equivalents of 4-ethylaniline were added, and the reaction was continued under TLC monitoring. After the reaction was complete, an appropriate amount of dichloromethane was added to the reaction solution, and the mixture was washed with water, dilute hydrochloric acid, saturated sodium bicarbonate aqueous solution, and saturated brine, respectively. The organic layer was slightly concentrated, and the solid precipitated. The solid was filtered and dried to obtain the target compound 1.

[0073] 1 H NMR (300 MHz, DMSO) δ 10.57 (s, 1H), 9.99 (s, 1H), 7.79 (dd, J =14.2, 8.4 Hz, 4H), 7.61 (d, J = 8.1 Hz, 2H), 7.18 (dd, J = 14.3, 8.3 Hz, 4H), 7.09 (d, J = 8.4 Hz, 2H), 3.80 (s, 3H), 2.58 (t, J = 7.7 Hz, 2H), 1.17 (t, J= 7.5 Hz, 3H). 13C NMR (75 MHz, DMSO) δ 164.59, 162.59, 140.88, 138.96,136.79, 130.91, 129.83, 128.95, 128.95, 128.87, 128.87, 127.73, 127.73,120.39, 120.39, 118.15, 118.15, 114.49, 114.49, 55.64, 27.59, 15.66.HRMS(ESI, m / z) calculated for [C 22 H 22 N2O4S+H] + : 411.1373; found: 411.1358.

[0074] Example 2: 4-((4-fluorophenyl)sulfonamido)-N-(4-(trifluoromethyl)phenyl)benzamide (compound 2)

[0075]

[0076] Compound 2 was prepared in accordance with Example 1, except that p-methoxybenzenesulfonyl chloride and 4-ethylaniline were replaced with p-fluorobenzenesulfonyl chloride and 4-trifluoromethylaniline, respectively, in the preparation of Compound 1.

[0077] 1 H NMR (400 MHz, DMSO) δ 10.83 (s, 1H), 10.45 (s, 1H), 8.02-7.82 (m,6H), 7.71 (d, J = 8.2 Hz, 2H), 7.49-7.40 (m, 2H), 7.28-7.21 (m, 2H). 13 C NMR(126 MHz, DMSO) δ 165.45, 165.27, 163.45, 142.86, 141.04, 135.74, 135.72,129.88, 129.81, 129.51, 129.28, 125.89, 125.86, 125.83, 125.80, 125.48,123.62, 123.37, 120.08, 118.52, 116.73, 116.55. HRMS (ESI, m / z) calculatedfor [C 20 H 14 F4N2O3S+H] + : 439.0734; found: 439.0797.

[0078] Example 3: N-(4-ethylphenyl)-4-((4-fluorophenyl)sulfonamido)benzamide (compound 3)

[0079]

[0080] Compound 3 was prepared in accordance with Example 1, except that p-methoxybenzenesulfonyl chloride and 4-ethylaniline were replaced with p-fluorobenzenesulfonyl chloride and 4-ethylaniline, respectively, in the preparation of Compound 1.

[0081] 1 H NMR (400 MHz, DMSO) δ 10.76 (s, 1H), 10.04 (s, 1H), 8.01-7.72 (m,4H), 7.62 (d, J = 8.0 Hz, 2H), 7.51-7.32 (m, 2H), 7.30-7.01 (m, 4H), 2.57 (q,J = 7.5 Hz, 2H), 1.17 (t, J = 7.5 Hz, 3H). 13 C NMR (126 MHz, DMSO) δ 165.44,164.55, 163.43, 140.44, 139.01, 136.77, 135.65, 130.28, 129.84, 129.77,128.97, 128.97, 127.75, 127.75, 120.40, 120.40, 118.58, 116.72, 116.54,27.60, 15.67. HRMS (ESI, m / z) calculated for [C 21 H 19 FN2O3S+H] + : 399.1173; found: 399.1161.

[0082] Example 4: 4-((4-methoxyphenyl)sulfonamido)-N-(3-(trifluoromethyl)phenyl)benzamide (compound 4)

[0083]

[0084] Compound 4 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with 3-trifluoromethylaniline.

[0085] 1H NMR (400 MHz, DMSO) δ 10.65 (s, 1H), 10.39 (s, 1H), 8.20 (d, J =2.1 Hz, 1H), 8.03-7.96 (m, 1H), 7.89-7.82 (m, 2H), 7.82-7.74 (m, 2H), 7.58(t, J = 8.0 Hz, 1H), 7.47- 7.40 (m, 1H), 7.27-7.19 (m, 2H), 7.14-7.05 (m, 2H), 3.81 (s, 3H). 13 C NMR (75 MHz, CDCl3) δ 165.14, 162.59, 141.30, 139.94,130.84, 129.77, 129.77, 129.09, 129.04, 129.04, 128.93, 128.93, 123.61,119.76, 118.04, 118.04, 116.21, 116.15, 114.49, 114.49, 55.63. HRMS (ESI, m / z) calculated for [C 21 H 17 F3N2O4S+H] + : 451.0934; found: 451.0910.

[0086] Example 5: 4-((4-fluorophenyl)sulfonamido)-N-(o-tolyl)benzamide (Compound 5)

[0087]

[0088] Compound 5 was prepared in accordance with Example 1, except that p-methoxybenzenesulfonyl chloride and 4-ethylaniline were replaced with p-fluorobenzenesulfonyl chloride and 2-methylaniline, respectively, in the preparation of Compound 1.

[0089] 1 H NMR (400 MHz, DMSO) δ 10.80 (s, 1H), 9.76 (s, 1H), 8.02-7.77 (m,4H), 7.53-7.37 (m, 2H), 7.35-7.08 (m, 6H), 2.20 (s, 3H). 13C NMR (126 MHz, Acetone) δ 165.39, 164.49, 163.38, 140.55, 136.39, 135.74, 133.63, 130.20,129.82, 129.74, 129.72, 128.94, 126.49, 125.88, 125.83, 118.62, 118.62,116.65, 116.47, 17.84. HRMS (ESI, m / z) calculated for [C 20 H 17 FN2O3S+H] + :385.1017; found: 385.0997.

[0090] Example 6: N-cyclohexyl-4-((4-methoxyphenyl)sulfonamide)benzamide (compound 6)

[0091]

[0092] Compound 6 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with cyclohexylamine.

[0093] 1 H NMR (400 MHz, DMSO) δ 10.49 (s, 1H), 8.05 (d, J = 8.0 Hz, 1H), 7.82 – 7.53 (m, 4H), 7.17-6.98 (m, 4H), 3.79 (s, 3H), 3.69 (s, 1H), 1.87-1.53(m, 5H), 1.27-1.23 (m, 5H). 13 C NMR (75 MHz, CDCl3) δ 164.59, 162.51, 140.32,130.85, 129.88, 128.88, 128.88, 128.40, 128.40, 118.18, 118.18, 114.41,114.41, 55.60, 48.19, 32.36, 32.36, 25.21, 24.87, 24.87. HRMS (ESI, m / z)calculated for [C 20 H 24 N2O4S+H] + : 389.1530; found: 389.1513.

[0094] Example 7: 4-((4-methoxyphenyl)sulfonamido)-N-(o-tolyl)benzamide (compound 7)

[0095]

[0096] Compound 7 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with 2-methylaniline.

[0097] 1 H NMR (400 MHz, DMSO) δ 10.60 (s, 1H), 9.71 (s, 1H), 7.85 (d, J =8.7 Hz, 2H), 7.81-7.73 (m, 2H), 7.32-7.06 (m, 8H), 3.81 (s, 3H), 2.19 (s,3H). 13 C NMR (75 MHz, CDCl3) δ 164.47, 162.56, 140.90, 136.37, 133.56, 130.87,130.20, 129.30, 128.91, 128.91, 128.86, 128.86, 126.43, 125.89, 125.81,118.10, 118.10, 114.48, 114.48, 55.61, 17.83. HRMS (ESI, m / z) calculated for[C 21 H 20 N2O4S+H] + : 397.1217; found: 397.1190.

[0098] Example 8: N-(2-ethylphenyl)-4-((4-methoxyphenyl)sulfonamide)benzamide (compound 8)

[0099]

[0100] Compound 8 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with 2-ethylaniline.

[0101] 1H NMR (300 MHz, DMSO) δ 10.58 (s, 1H), 9.71 (s, 1H), 7.81 (dd, J =19.8, 8.4 Hz, 4H), 7.32-7.04 (m, 8H), 3.81 (s, 3H), 2.71- 2.55 (m, 2H), 1.10(t, J = 7.4 Hz, 3H). 13 C NMR (75 MHz, CDCl3) δ 164.86, 162.57, 140.90, 139.71,135.74, 130.89, 129.30, 128.92, 128.92, 128.83, 128.83, 128.37, 127.43,126.34, 125.91, 118.12, 118.12, 114.50, 114.50, 55.62, 23.86, 14.04. HRMS(ESI, m / z) calculated for [C 22 H 22 N2O4S+H] + : 411.1373; found: 411.1354.

[0102] Example 9: N-cyclohexyl-4-((4-fluorophenyl)sulfonamide)benzamide (compound 9)

[0103]

[0104] Compound 9 was prepared in accordance with Example 1, except that p-methoxybenzenesulfonyl chloride and 4-ethylaniline were replaced with p-fluorobenzenesulfonyl chloride and cyclohexylamine, respectively, in the preparation of Compound 1.

[0105] 1 H NMR (300 MHz, DMSO) δ 10.60 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.91-7.78 (m, 2H), 7.71 (d, J = 7.4 Hz, 2H), 7.51-7.34 (m, 2H), 7.14 (d, J =7.6 Hz, 2H), 3.70 (s, 1H), 1.88-1.50 (m, 5H), 1.33-1.08 (m, 5H). 13C NMR (75MHz, CDCl3) δ 166.01, 164.53, 162.67, 139.87, 135.58, 130.32, 129.78, 129.66,128.48, 118.64, 118.64, 116.67, 116.37, 48.20, 32.35, 32.35, 25.20, 24.86,24.86. HRMS (ESI, m / z) calculated for [C 19 H 21 FN2O3S+H] + : 377.1330; found:377.1305.

[0106] Example 10: Ethyl 4-(4-(4-methoxyphenyl)sulfonamido)benzamido)benzoate (Compound 10)

[0107]

[0108] Compound 10 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with ethyl 4-aminobenzoate.

[0109] 1 H NMR (300 MHz, DMSO) δ 10.63 (s, 1H), 10.39 (s, 1H), 8.04-7.71 (m,8H), 7.23 (d, J = 8.0 Hz, 2H), 7.09 (d, J = 8.4 Hz, 2H), 4.40- 4.21 (m, 2H),3.81 (s, 3H), 1.32 (t, J = 6.9 Hz, 3H). 13 C NMR (75 MHz, CDCl3) δ 165.30,165.18, 162.59, 143.60, 141.28, 130.84, 129.98, 129.98, 129.23, 129.13,129.13, 128.93, 128.93, 124.41, 119.40, 119.40, 118.01, 118.01, 114.50,114.50, 60.38, 55.63, 14.17. HRMS (ESI, m / z) calculated for [C 23 H 22 N2O6S+H] +:455.1271; found: 455.1251

[0110] Example 11: N-(furan-2-ylmethyl)-4-((4-methoxyphenyl)sulfonamide)benzoyl (compound 11)

[0111]

[0112] Compound 11 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with 2-furanmethylamine.

[0113] 1 H NMR (400 MHz, DMSO) δ 10.51 (s, 1H), 8.80 (t, J = 5.7 Hz, 1H), 7.79-7.71 (m, 4H), 7.56 (dd, J = 1.8, 0.9 Hz, 1H), 7.20-7.13 (m, 2H), 7.12-7.04 (m, 2H), 6.39 (dd, J = 3.3, 1.9 Hz, 1H), 6.25 (d, J = 3.2 Hz, 1H), 4.43(d, J = 5.6 Hz, 2H), 3.81 (s, 3H). 13 C NMR (75 MHz, DMSO) δ 165.30, 162.51,152.38, 141.83, 140.65, 130.86, 129.03, 128.83, 128.83, 128.43, 128.43,118.17, 118.17, 114.40, 114.40, 110.34, 106.68, 55.56, 35.88. HRMS (ESI, m / z)calculated for [C 19 H 18 N2O5S+H] + : 387.1010; found: 387.1031.

[0114] Example 12: N-(4-(diethylamino)phenyl)-4-((4-methoxyphenyl)sulfonamide)benzamide (compound 12)

[0115]

[0116] Compound 12 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with 4-diethylaminoaniline.

[0117] 1 H NMR (300 MHz, DMSO) δ 10.61 (s, 1H), 9.82 (s, 1H), 7.89- 7.71 (m,4H), 7.49 (d, J = 8.9 Hz, 2H), 7.21 (d, J = 8.4 Hz, 2H), 7.11 (d, J = 8.7 Hz, 2H), 6.65 (d, J = 8.7 Hz, 2H), 3.82 (s, 3H), 3.35- 3.21 (m, 4H), 1.09 (t, J =6.9 Hz, 6H). 13 C NMR (75 MHz, DMSO) δ 163.89, 162.52, 144.21, 140.58, 130.90,130.08, 128.90 (2C), 128.62 (2C), 127.73, 122.18 (2C), 118.16 (2C), 114.43(2C), 111.71 (2C), 55.60, 43.71 (2C), 12.35 (2C). HRMS (ESI, m / z) calculated for [C 24 H 27 N3O4S+H] + : 454.1795; found: 454.1806.

[0118] Example 13: Ethyl 2-(4-(4-methoxyphenyl)sulfonamido)benzoamide)thiazole-5-carboxylate (Compound 13)

[0119]

[0120] Compound 13 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with ethyl 2-aminothiazole-5-carboxylate.

[0121] 1H NMR (400 MHz, DMSO) δ 12.89 (s, 1H), 10.76 (s, 1H), 8.21 (s, 1H), 8.00 (d, J = 8.8 Hz, 2H), 7.87- 7.76 (m, 2H), 7.24 (d, J = 8.8 Hz, 2H), 7.11(dd, J = 11.4, 8.7 Hz, 2H), 4.30 (q, J = 7.1 Hz, 2H), 3.81 (s, 3H), 1.31 (t,J = 7.1 Hz, 3H). 13 C NMR (75 MHz, DMSO) δ 164.88, 162.97, 162.66, 161.42,144.95, 142.33, 130.79, 129.80, 128.93, 127.32, 125.89, 121.33, 118.94,117.76, 114.54, 114.37, 60.80, 55.62, 14.13. HRMS (ESI, m / z) calculated for[C 20 H 19 N3O6S2+H] + : 462.0789; found: 462.0809.

[0122] Example 14: N-Isopropyl-4-((4-methoxyphenyl)sulfonamide)benzamide (Compound 14)

[0123]

[0124] Compound 14 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with isopropylamine.

[0125] 1 H NMR (400 MHz, DMSO) δ 10.43 (s, 1H), 8.01 (d, J = 7.7 Hz, 1H), 7.79-7.58 (m, 4H), 7.17-6.98 (m, 4H), 4.04 (d, J = 6.7 Hz, 1H), 3.85- 3.72(m, 3H), 1.12 (d, J = 6.6 Hz, 6H). 13C NMR (126 MHz, DMSO) δ 169.46, 164.67,162.54, 162.52, 140.41, 138.64, 132.31, 130.97, 130.88, 129.84, 128.92,128.88, 128.39, 127.36, 118.95, 118.21, 114.43, 114.41, 55.61, 40.85, 22.29.HRMS (ESI, m / z) calculated for [C 17 H 20 N2O4S+H] + : 349.1217; found: 349.1216.

[0126] Example 15: Methyl 2-(4-(4-methoxyphenyl)sulfonamido)benzamido)thiazole-4-carboxylate (Compound 15)

[0127]

[0128] Compound 15 was prepared in accordance with Example 1, except that 4-ethylaniline in the preparation of Compound 1 was replaced with methyl 2-aminothiazole-4-carboxylate.

[0129] 1 H NMR (400 MHz, DMSO) δ 12.83 (s, 1H), 10.74 (s, 1H), 8.10 (s, 1H), 8.02 (d, J = 8.8 Hz, 2H), 7.82-7.74 (m, 2H), 7.23 (dd, J = 8.6, 3.9 Hz, 2H),7.13-7.07 (m, 2H), 3.82 (d, J = 10.2 Hz, 6H). 13 C NMR (75 MHz, DMSO) δ 164.60,162.65, 161.41, 158.74, 142.17, 140.73, 130.81, 129.61, 128.90, 128.83,127.31, 125.97, 123.04, 118.93, 117.77, 114.55, 114.37, 55.61, 51.80. HRMS(ESI, m / z) calculated for [C 19 H 17 N3O6S2+H] +: 448.0632; found: 448.0650.

[0130] Example 16: N-cyclohexyl-4-(2,4,6-trimethylphenyl)sulfonamide (compound 16)

[0131]

[0132] Compound 16 was prepared in accordance with Example 1, except that p-methoxybenzenesulfonyl chloride and 4-ethylaniline were replaced with 2,4,6-trimethylbenzenesulfonyl chloride and cyclohexylamine, respectively.

[0133] 1 H NMR (400 MHz, DMSO) δ 10.56 (s, 1H), 7.99 (d, J = 7.5 Hz, 1H), 7.68 (d, J = 8.7 Hz, 2H), 7.07-6.90 (m, 4H), 3.75-3.61 (m, 1H), 2.59 (s, 6H),2.22 (s, 3H), 1.84-1.52 (m, 5H), 1.30-1.02 (m, 5H). 13 C NMR (126 MHz, Acetone)δ 164.62, 142.29, 140.20, 138.69, 138.69, 133.50, 131.84, 131.84, 129.40,128.46, 128.64, 117.08, 117.08, 48.18, 32.38, 32.38, 25.22, 24.87, 24.87,22.35, 22.35, 20.33. HRMS (ESI, m / z) calculated for [C 22 H 28 N2O3S+H] + : 401.1893; found: 401.1878.

[0134] Example 17: N-(4-(diethylamino)phenyl)-4-((2,4,6-trimethylphenyl)sulfonamido)benzamide (compound 17)

[0135]

[0136] Compound 17 was prepared in accordance with Example 1, except that p-methoxybenzenesulfonyl chloride and 4-ethylaniline in the preparation of Compound 1 were replaced with 2,4,6-trimethylbenzenesulfonyl chloride and 4-diethylaminoaniline, respectively.

[0137] 1 H NMR (400 MHz, DMSO) δ 10.63 (s, 1H), 9.74 (s, 1H), 7.78 (s, 2H), 7.46 (s, 2H), 7.04 (s, 4H), 6.64 (s, 2H), 3.34 (d, J = 2.0 Hz, 4H), 2.61 (s,6H), 2.23 (s, 3H), 1.19- 1.00 (m, 6H). 13 C NMR (75 MHz, DMSO) δ 163.90,144.22, 142.31, 140.41, 138.69 (2C), 133.56, 131.86 (2C), 129.66, 128.68(2C), 127.80, 122.12 (2C), 117.08 (2C), 111.77 (2C), 43.73 (2C), 22.35 (2C),20.33, 12.36 (2C). HRMS (ESI, m / z) calculated for [C 26 H 31 N3O3S+H] + : 466.2159; found: 466.2155.

[0138] Example 18: N-(furan-2-ylmethyl)-4-((2,4,6-trimethylphenyl)sulfonamide)benzamide (compound 18)

[0139]

[0140] Compound 18 was prepared in accordance with Example 1, except that p-methoxybenzenesulfonyl chloride and 4-ethylaniline were replaced with 2,4,6-trimethylbenzenesulfonyl chloride and 2-furanmethylamine, respectively.

[0141] 1H NMR (400 MHz, DMSO) δ 10.60 (s, 1H), 8.76 (t, J = 5.8 Hz, 1H), 7.71 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 1.9 Hz, 1H), 7.01 (d, J = 7.8 Hz, 4H), 6.37 (dd, J = 3.2, 1.9 Hz, 1H), 6.23 (d, J = 3.2 Hz, 1H), 4.40 (d, J = 5.7Hz, 2H), 2.59 (s, 6H), 2.22 (s, 3H). 13 C NMR (75 MHz, DMSO) δ 166.35, 162.78,153.43, 141.43, 140.85, 131.69, 129.95, 129.03, 129.03, 128.26, 128.26,119.03, 119.03, 115.13, 115.13, 111.63, 106.98, 36.89, 22.41, 22.41, 20.97.HRMS (ESI, m / z) calculated for [C 21 H 22 N2O4S+H] + : 399.1373; found: 399.1383.

[0142] Example 19: 2-((4-methoxyphenyl)sulfonamido)-N-(4-(trifluoromethyl)phenyl)thiazole-4-carboxamide (Compound 19)

[0143]

[0144] (1) Synthetic intermediate 19-M1

[0145] 2 mmol of methyl 2-aminothiazolyl-4-carboxylate was dissolved in dichloromethane, and 7.5 equivalents of pyridine were added. The mixture was stirred and the reaction was carried out. 2.4 mmol of p-methoxysulfonyl chloride dissolved in dichloromethane was slowly added dropwise to the reaction solution. The reaction was carried out at 50 °C and monitored by TLC. After the reaction was completed, the reaction solution was added to a saturated sodium bicarbonate solution and extracted three times with dichloromethane. The dichloromethane layers were combined and washed with hydrochloric acid solution and water, respectively. The organic layer was concentrated to obtain intermediate 19-M1.

[0146] (2) Synthetic intermediate 19-M2

[0147] 19-M1 was dissolved in methanol, and 4 times the volume of lithium hydroxide aqueous solution was added. The reaction was stirred at 50°C. The hydrolysis reaction was monitored by TLC until it was complete. The methanol was removed by concentration, and the pH was adjusted to acidic with dilute hydrochloric acid. A large amount of precipitate was formed. The precipitate was filtered, washed with water, and dried to obtain intermediate 19-M2.

[0148] (3) Synthesis of 2-((4-methoxyphenyl)sulfonamido)-N-(4-(trifluoromethyl)phenyl)thiazole-4-carboxamide (compound 19)

[0149]

[0150] 19-M2 was completely dissolved in dry DMF, and 1.5 equivalents of HATU and 2 equivalents of triethylamine were added. The mixture was magnetically stirred and activated for 30 min. Then, 1.2 equivalents of 4-trifluoromethylaniline were added, and the reaction was continued under TLC monitoring. After the reaction was complete, an appropriate amount of dichloromethane was added to the reaction solution, and the mixture was washed with water, dilute hydrochloric acid, saturated sodium bicarbonate aqueous solution, and saturated brine, respectively. The organic layer was slightly concentrated, and the solid precipitated. The solid was filtered and dried to obtain the target compound 19.

[0151] 1 H NMR (300 MHz, DMSO) δ 12.97 (s, 1H), 10.53 (s, 1H), 8.15- 7.58 (m,7H), 7.07 (d, J = 8.8 Hz, 2H), 3.80 (s, 3H). 13 C NMR (75 MHz, DMSO) δ 162.15,156.43, 141.76, 133.58, 128.00, 128.00, 126.12, 126.07, 126.02, 125.97,124.25, 123.83, 122.44, 120.00, 120.00, 114.17, 114.17, 55.58, 54.85. HRMS(ESI, m / z) calculated for [C 18 H 14 F3N3O4S2+H] + : 458.0451; found: 458.0484.

[0152] Example 20: 2-((4-methoxyphenyl)sulfonamido)-N-(o-tolyl)thiazole-4-carboxamide (Compound 20)

[0153]

[0154] Compound 20 was prepared according to Example 19, except that 4-trifluoromethylaniline was replaced with 2-methylaniline in the preparation of Compound 19.

[0155] 1 H NMR (300 MHz, DMSO) δ 12.99 (s, 1H), 9.81 (s, 1H), 7.92- 7.56 (m,3H), 7.42- 7.00 (m, 5H), 3.80 (s, 3H), 2.18 (s, 3H). 13 C NMR (75 MHz, DMSO) δ166.29, 162.13, 156.39, 135.12, 133.60, 132.76, 130.40, 128.06(2C), 126.13(2C), 126.10, 125.55, 114.15(2C), 113.41, 55.58, 17.64. HRMS (ESI, m / z)calculated for [C 18 H 17 N3O4S2+H] + : 404.0733; found: 404.0759.

[0156] Example 21: N-(4-ethylphenyl)-2-((4-methoxyphenyl)sulfonamido)thiazole-5-carboxamide (Compound 21)

[0157]

[0158] Compound 21 was prepared in accordance with Example 19, except that methyl 2-aminothiazole-4-carboxylate and 4-trifluoromethylaniline were replaced with ethyl 2-aminothiazole-5-carboxylate and 4-ethylaniline, respectively. 1 HNMR (300 MHz, DMSO) δ 12.95 (s, 1H), 10.05 (s, 1H), 8.09 (s, 1H), 7.75 (d, J= 8.8 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 7.11 (dd, J = 30.1, 8.6 Hz, 4H), 3.80 (s, 3H), 2.55 (q, J = 7.6 Hz, 2H), 1.15 (t, J = 7.6 Hz, 3H). 13C NMR (75MHz, DMSO) δ 167.78, 162.17, 157.75, 139.38, 135.87, 133.44, 127.99, 127.99,127.94, 127.94, 127.63, 120.40, 120.16, 120.16, 114.18, 114.18, 55.57, 27.55,15.56. HRMS (ESI, m / z) calculated for [C 19 H 19 N3O4S2+H] + : 418.0890; found:418.0915.

[0159] Example 22: Ethyl 4-(2-((4-methoxyphenyl)sulfonamido)thiazole-5-carbamate)benzoate (Compound 22)

[0160]

[0161] Compound 22 was prepared in accordance with Example 19, except that methyl 2-aminothiazole-4-carboxylate and 4-trifluoromethylaniline were replaced with ethyl 2-aminothiazole-5-carboxylate and ethyl 4-aminobenzoate, respectively.

[0162] 1 H NMR (300 MHz, DMSO) δ 13.10 (s, 1H), 10.65 (s, 1H), 8.41 (s, 1H), 8.09- 7.58 (m, 6H), 7.06 (d, J = 8.8 Hz, 2H), 4.28 (q, J = 7.0 Hz, 2H), 3.80(s, 3H), 1.30 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, DMSO) δ 166.18, 165.94,163.82, 161.81, 142.25, 138.08, 132.01, 130.16, 130.16, 126.34, 126.34,125.76, 121.16, 118.69, 118.69, 114.62, 114.62, 59.92, 55.84, 14.06. HRMS(ESI, m / z) calculated for [C 20 H 19 N3O6S2+H] +: 462.0789; found: 462.0832

[0163] Example 23: 2-((4-methoxyphenyl)sulfonamido)-N-(4-(trifluoromethyl)phenyl)thiazole-5-carboxamide (Compound 23)

[0164]

[0165] Compound 23 was prepared in accordance with Example 19, except that methyl 2-aminothiazole-4-carboxylate was replaced with ethyl 2-aminothiazole-5-carboxylate in the preparation of Compound 19.

[0166] 1 H NMR (300 MHz, DMSO) δ 13.06 (s, 1H), 10.40 (s, 1H), 8.17 (s, 1H), 7.85 (d, J = 8.5 Hz, 2H), 7.73 (dd, J = 16.0, 8.7 Hz, 4H), 7.06 (d, J = 8.8Hz, 2H), 3.80 (s, 3H). 13 C NMR (126 MHz, DMSO) δ 168.09, 162.16, 158.52,142.03, 133.55, 129.59, 128.02, 127.53, 126.12, 126.09, 126.05, 126.02,125.37, 123.86, 123.61, 123.36, 123.22, 119.78, 119.69, 114.17, 55.57. HRMS(ESI, m / z) calculated for [C 18 H 14 F3N3O4S2+H] + : 458.0451; found: 458.0488.

[0167] Example 24: N-isopropyl-2-((4-methoxyphenyl)sulfonamido)thiazole-5-carboxamide (Compound 24)

[0168]

[0169] The preparation of compound 24 is based on Example 19, except that methyl 2-aminothiazole-4-carboxylate and 4-trifluoromethylaniline in the preparation of compound 19 are replaced with ethyl 2-aminothiazole-5-carboxylate and isopropylamine, respectively.

[0170] 1 H NMR (400 MHz, DMSO) δ 12.88 (s, 1H), 8.29 (s, 1H), 8.06-7.89 (m,1H), 7.84-7.68 (m, 2H), 7.16-6.97 (m, 2H), 3.97 (dd, J = 13.3, 6.8 Hz, 1H), 3.82 (s, 3H), 1.34-1.05 (m, 6H). 13 C NMR (101 MHz, DMSO) δ 168.44, 162.59,158.93, 134.21, 128.49, 128.49, 127.39, 121.23, 114.65, 114.65, 56.09, 41.68,22.75,22.75. HRMS (ESI, m / z) calculated for [C 14 H 17 N3O4S2+H] + : 356.0733; found:356.0684.

[0171] Example 25: 2-((4-methoxyphenyl)sulfonamido)-N-(3-(trifluoromethyl)phenyl)thiazole-4-carboxamide (Compound 25)

[0172]

[0173] Compound 25 was prepared according to Example 19, except that 4-trifluoromethylaniline was replaced with 3-trifluoromethylaniline in the preparation of Compound 19.

[0174] 1 H NMR (300 MHz, DMSO) δ 13.11(s,1H), 10.61 (s, 1H), 8.15 (s, 1H),7.96 (d, J = 8.3 Hz, 1H), 7.91- 7.72 (m, 3H), 7.64 (t, J = 7.9 Hz, 1H), 7.51(d, J = 7.7 Hz, 1H), 7.12 (d, J = 8.5 Hz, 2H), 3.85 (s, 3H). 13C NMR (76 MHz, DMSO) δ 162.13, 156.36, 138.88, 130.01, 129.62, 129.20, 127.98 (2C), 123.62,120.40, 120.35, 116.19, 116.14, 114.14, 114.06 (2C), 55.55. HRMS (ESI, m / z)calculated for [C 18 H 14 F3N3O4S2+H] + : 458.0451; found: 458.0475.

[0175] Example 26: Ethyl 4-(2-((4-methoxyphenyl)sulfonamido)thiazole-4-carbamate)benzoate (Compound 26)

[0176]

[0177] Compound 26 was prepared according to Example 19, except that 4-trifluoromethylaniline in the preparation of Compound 19 was replaced with ethyl 4-aminobenzoate.

[0178] 1 H NMR (400 MHz, DMSO) δ 13.03 (s, 1H), 10.52 (s, 1H), 8.06-7.69 (m,7H), 7.16- 7.00 (m, 2H), 4.31 (q, J = 7.1 Hz, 2H), 3.83 (s, 3H), 1.30 (t, J =7.1 Hz, 3H). 13 C NMR (126 MHz, DMSO) δ 165.25, 162.14, 156.41, 142.52, 133.69,130.17 (3C), 128.03 (2C), 125.04, 119.47 (3C), 114.18 (3C), 60.52, 55.60,14.17. HRMS (ESI, m / z) calculated for [C 20 H 19 N3O6S2+H] + : 462.0788; found:462.0727.

[0179] Example 27: Methyl 2-(2-((4-methoxyphenyl)sulfonamide)thiazole-4-carboxamido)thiazole-4-carboxylic acid (Compound 27)

[0180]

[0181] Compound 27 was prepared in accordance with Example 19, except that 4-trifluoromethylaniline in the preparation of Compound 19 was replaced with methyl 2-aminothiazole 4-carboxylate.

[0182] 1 H NMR (400 MHz, DMSO) δ 8.11 (s, 1H), 7.76-7.69 (m, 2H), 7.59 (s,1H), 7.01-6.94 (m, 2H), 3.84 (s, 3H), 3.78 (s, 3H). 13 C NMR (101 MHz, DMSO) δ169.87, 162.04, 161.19, 160.28, 158.42, 141.25, 137.78, 128.34, 128.34,123.76, 116.69, 113.82, 113.82, 55.84, 52.47. HRMS (ESI, m / z) calculated for[C 16 H 14 N4O6S3+H] + : 455.0149; found: 455.0161.

[0183] Example 28: N-((1s,3s)-adamantane-1-yl)-2-((4-methoxyphenyl)sulfonamido)thiazole-4-carboxamide (Compound 28)

[0184]

[0185] Compound 28 was prepared according to Example 19, except that 4-trifluoromethylaniline was replaced with 1-adamantaneamine in the preparation of Compound 19.

[0186] 1 H NMR (400 MHz, DMSO) δ 12.79 (s, 1H), 7.74 (d, J = 8.7 Hz, 2H), 7.65 (s, 1H), 7.53 (s, 1H), 7.07 (d, J = 8.8 Hz, 2H), 3.82 (s, 3H), 2.04-1.99(m, 9H), 1.63(s, 6H). 13C NMR (75 MHz, DMSO) δ 162.09, 156.44, 135.14, 133.64,127.97, 127.97, 114.12, 114.14, 114.14, 112.12, 55.57, 51.89, 40.64, 40.64,40.64, 35.86, 35.86, 28.76, 28.76, 28.76. HRMS (ESI, m / z) calculated for[C 21 H 25 N3O4S2+H] + : 448.1359; found: 448.1296.

[0187] Example 29: N-(furan-2-ylmethyl)-2-((4-methoxyphenyl)sulfonamido)thiazole-4-carboxamide (Compound 29)

[0188]

[0189] Compound 29 was prepared in accordance with Example 19, except that 4-trifluoromethylaniline was replaced with 2-furanmethylamine in the preparation of Compound 19.

[0190] 1 H NMR (400 MHz, DMSO) δ 12.86 (s, 1H), 8.92 (s, 1H), 7.75 (d, J =8.8 Hz, 2H), 7.61- 7.50 (m, 2H), 7.07 (d, J = 8.9 Hz, 2H), 6.40 (dd, J = 3.1,1.9 Hz, 1H), 6.29 (d, J = 3.0 Hz, 1H), 4.40 (d, J = 5.6 Hz, 2H), 3.82 (s,3H). 13 C NMR (75 MHz, DMSO) δ 165.14, 162.79, 158.04, 143.83, 142.91, 140.13,131.07, 125.13, 125.13, 112.63, 112.63, 111.46, 109.37, 109.56, 55.78, 36.44.HRMS (ESI, m / z) calculated for [C 16 H 15 N3O5S2+H] + : 394.0526; found: 394.0552.

[0191] Example 30: N-isopropyl-2-((4-methoxyphenyl)sulfonamide)thiazole-4-carboxamide (Compound 30)

[0192]

[0193] The preparation of compound 30 is based on Example 19, except that 4-trifluoromethylaniline in the preparation of compound 19 is replaced with isopropylamine.

[0194] 1 H NMR (400 MHz, DMSO) δ 12.87 (s, 1H), 8.26 (s, 1H), 7.87- 7.66 (m,2H), 7.50 (s, 1H), 7.07 (dd, J = 8.8, 1.6 Hz, 2H), 3.96 (dq, J = 13.1, 6.7Hz, 1H), 3.82 (d, J = 1.7 Hz, 3H), 1.12 (dd, J = 6.6, 1.5 Hz, 6H). 13 C NMR (75MHz, DMSO) δ 162.07, 156.41, 133.77, 127.95, 127.95, 114.11, 114.11, 114.11,114.11, 111.56, 55.58, 41.10, 22.05, 22.05. HRMS (ESI, m / z) calculated for[C 14 H 17 N3O4S2+H] + : 356.0733; found: 356.0682.

[0195] Example 31: N-(4-(diethylamino)phenyl)-2-((4-methoxyphenyl)sulfonamido)thiazole-4-carboxamide (Compound 31)

[0196]

[0197] The preparation of compound 31 is based on Example 19, except that 4-trifluoromethylaniline in the preparation of compound 19 is replaced with 4-diethylaminoaniline.

[0198] 1H NMR (400 MHz, DMSO) δ 12.93 (s, 1H), 9.79 (s, 1H), 7.75 (d, J =8.8 Hz, 2H), 7.54 (s, 1H), 7.45 (d, J = 8.9 Hz, 2H), 7.05 (d, J = 8.7 Hz,2H), 6.65 (d, J = 8.9 Hz, 2H), 3.81 (s, 3H), 3.33 (d, J = 7.0 Hz, 4H), 1.07(t, J = 7.0 Hz, 6H). 13 C NMR (75 MHz, DMSO) δ 166.93, 162.00, 155.41, 144.16,133.95, 127.94, 127.94, 127.94, 126.92, 121.89, 121.89, 114.08, 114.08,114.08, 112.08, 112.08, 55.55, 44.00, 44.00, 12.27, 12.27. HRMS (ESI, m / z)calculated for [C 21 H 24 N4O4S2+H] + : 461.1312; found: 461.1321.

[0199] Example 32: N-(2-ethylphenyl)-2-((4-methoxyphenyl)sulfonamido)thiazole-4-carboxamide (Compound 32)

[0200]

[0201] Compound 32 was prepared according to Example 19, except that 4-trifluoromethylaniline was replaced with 2-ethylaniline in the preparation of Compound 19.

[0202] 1 H NMR (400 MHz, DMSO) δ 13.02 (s, 1H), 9.89 (s, 1H), 7.86-7.67 (m,3H), 7.39-7.19 (m, 4H), 7.10 (d, J = 8.9 Hz, 2H), 3.83 (s, 3H), 2.58 (q, J =7.5 Hz, 2H), 1.12 (t, J = 7.5 Hz, 3H). 13C NMR (75 MHz, DMSO) δ 167.81,162.20, 157.80, 139.43, 135.89, 133.47, 128.02, 128.02, 127.96, 127.96,127.67, 120.46, 120.21, 120.21, 114.21, 114.21, 55.59, 27.58, 15.58. HRMS(ESI, m / z) calculated for [C 19 H 19 N3O4S2+H] + : 418.0890; found: 418.0895.

[0203] Example 33: N-cyclohexyl-2-((4-methoxyphenyl)sulfonamido)thiazole-5-carboxamide (Compound 33)

[0204]

[0205] The preparation of compound 33 is based on Example 19, except that methyl 2-aminothiazole-4-carboxylate and 4-trifluoromethylaniline in the preparation of compound 19 are replaced with ethyl 2-aminothiazole-5-carboxylate and cyclohexylamine, respectively.

[0206] 1 H NMR (400 MHz, DMSO) δ 12.85 (s, 1H), 8.20 (d, J = 7.8 Hz, 1H), 7.94 (s, 1H), 7.82-7.67 (m, 2H), 7.22-7.00 (m, 2H), 3.82 (s, 3H), 3.64 (ddd,J = 11.1, 7.5, 3.5 Hz, 1H), 1.75 (dd,J = 29.6, 11.2 Hz, 5H), 1.25 (q, J =12.9 Hz, 5H). 13 C NMR (75 MHz, DMSO) δ 167.80, 162.09, 158.26, 133.58, 127.94, 127.94, 126.39, 120.68, 114.13, 114.13, 55.55, 48.36, 32.31, 32.31, 25.11,24.71, 24.71. HRMS (ESI, m / z) calculated for [C 17 H 21 N3O4S2+H]+ : 396.1046; found:396.1045.

[0207] Example 34: 2-((4-methoxyphenyl)sulfonamido)-N-(o-tolyl)thiazole-5-carboxamide (Compound 34)

[0208]

[0209] The preparation of compound 34 is based on Example 19, except that methyl 2-aminothiazole-4-carboxylate and 4-trifluoromethylaniline in the preparation of compound 19 are replaced with ethyl 2-aminothiazole-5-carboxylate and 2-methylaniline, respectively.

[0210] 1 H NMR (400 MHz, DMSO) δ 13.01 (s, 1H), 9.85 (s, 1H), 8.10 (s, 1H), 7.78 (d, J = 8.8 Hz, 2H), 7.32- 7.04 (m, 6H), 3.83 (s, 3H), 2.21 (s, 3H). 13 CNMR (75 MHz, DMSO) δ 167.85, 162.36, 157.63, 138.83, 135.81, 133.17, 128.32, 128.32, 127.76, 127.96, 127.67, 120.46, 120.01, 120.01, 114.35, 114.35,55.79, 16.78. HRMS (ESI, m / z) calculated for [C 18 H 17 N3O4S2+H] + : 404.0733; found:404.0738.

[0211] The sources of the various strains involved in the following experimental examples:

[0212] The standard strain of Klebsiella pneumoniae (K. pneumoniae subsp.) ATCC13883, Salmonella typhimurium (S. typhimurium) CGMCC1.1174, Streptococcus agalactiae (S. agalactiae) ATCC13813, Enterococcus faecalis (E. faealis) ATCC 29212, and Vibrio harveyi reporter strain BB170 (Vibrio harveyi ATCC BAA-1117) were purchased from Beijing Microbial Culture Collection Center (Beijing, China). Pseudomonas aeruginosa (P. aeruginosa) CGMCC1.50 and Staphylococcus aureus (S. aureus) ATCC25923 were purchased from the China General Microbiological Culture Collection Center (Beijing, China); Klebsiella pneumoniae KP31, a clinically isolated bacterium exhibiting high virulence and multidrug resistance characteristics after whole-genome sequencing, was collected from bacterial isolates from the First Affiliated Hospital of Huzhou University (S. Yi et al. Microbial Pathogenesis, 2026, 213, 108332).

[0213] Experimental Example 1: Antibacterial Activity Detection

[0214] This study evaluated the direct antibacterial activity of the compounds by determining their minimum inhibitory concentrations (MICs) against seven Gram-positive and Gram-negative bacteria. The tested strains included Klebsiella pneumoniae subsp. ATCC 13883, Salmonella typhimurium CGMCC 1.1174, Streptococcus agalactiae ATCC 13813, Enterococcus faecalis ATCC 29212, Pseudomonas aeruginosa CGMCC 1.50, and Staphylococcus aureus ATCC 25923, as well as a highly virulent, multidrug-resistant clinical isolate of Klebsiella pneumoniae (KP31).

[0215] In this experiment, the MIC (micron activity) of 34 compounds was determined using the two-fold broth dilution method. Kanamycin (Kan), streptomycin (Str), azithromycin (Azm), and chloramphenicol (Cm) were used as positive control drugs. All control drugs (Kan, Streptomycin, Azm, and Chloramphenicol) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. The antibacterial activity of the compounds was determined based on the measured MIC values, and the results are shown in Table 1.

[0216] Table 1. MIC values ​​of compounds 1-34 against 7 pathogenic strains

[0217]

[0218] The experimental results in Table 1 show that all compounds of this invention have MICs ≥ 512 μg / mL against the standard strain of Klebsiella pneumoniae (K. pneumoniaesubsp ATCC 13883), highly virulent multidrug-resistant clinical isolates of Klebsiella pneumoniae (KP31), and Pseudomonas aeruginosa (P. aeruginosa CGMCC 1.50), indicating that these compounds have no direct antibacterial activity against these three strains. For the remaining tested strains, including Streptococcus agalactiae (S. agalactiae ATCC 13813), Enterococcus faecalis (E. faealis ATCC 29212), Staphylococcus aureus (S. aureus ATCC 25923), and Salmonella typhimurium (S. typhimurium CGMCC 1.1174), most compounds also have MICs ≥ 512 μg / mL. Only a very few compounds have MICs of 256 μg / mL against specific strains, still falling into the category of having no significant antibacterial activity. The overall activity data fully confirm that the compounds of this invention do not have significant bacterial growth inhibition ability and are not subject to bacterial resistance screening pressure, which fully meets the core development premise that quorum sensing inhibitors (QSIs) have no direct antibacterial activity.

[0219] Experimental Example 2: Effect of Compounds on AI-2 Formation in Klebsiella pneumoniae KP31

[0220] AI-2 signaling molecules are a core component of the quorum sensing (QS) system in Klebsiella pneumoniae, playing a pivotal role in coordinating key physiological processes such as biofilm formation, virulence factor secretion, and bacterial pathogenicity. This study characterized the QS inhibitory effect of the compounds in this invention by determining the effects of all 34 target compounds on the AI-2 signaling molecule production level of highly virulent, multidrug-resistant Klebsiella pneumoniae strain KP31. The bioluminescence assay of Vibrio harveyi reporter strain BB170 was used to evaluate the effect of each compound on the AI-2 signaling molecule.

[0221] The specific experimental method is as follows: Highly virulent, multidrug-resistant Klebsiella pneumoniae KP31 bacterial suspension was co-cultured with each target compound at a final concentration of 256 µg / mL for 8 h. Cell-free supernatants were then collected by centrifugation (1000 rpm, 15 min, 4°C) and filtered through a 0.22 μm filter for sterilization. The resulting supernatant was mixed with Vibrio harveyi BB170 bacterial suspension at a volume ratio of 1:9 and incubated for 1 h. Bioluminescence intensity was measured using a SpectraMax iD3 multi-functional microplate reader. AI-2 activity was expressed as relative luminescence value (experimental group / control group), and all experiments were performed in triplicate.

[0222] Experimental results are as follows Figure 1 As shown, compounds 1, 4, 6, 8, 9, 10, 11, 13, 16, 31, and 32 significantly inhibit the generation of AI-2 signaling molecules, with compounds 1, 6, and 13 exhibiting the strongest inhibitory effects, achieving inhibition rates of 16%-18% on AI-2 signal generation.

[0223] Experimental Example 3: Effect of Compounds on Biofilm Formation of Klebsiella pneumoniae KP31

[0224] To further verify the quorum sensing inhibitory activity of the compounds of the present invention, this experiment characterized the QS inhibitory effect of the compounds of the present invention by measuring the effect of all 34 target compounds on the formation of Klebsiella pneumoniae KP31 biofilm.

[0225] The specific experimental method is as follows: Dilute the overnight cultured Klebsiella pneumoniae KP31 bacterial suspension to OD200. 600 ≈ 0.05, then add 150 µL to each well of a 96-well plate. Next, add the target compound to each well to a final concentration of 128 μg / mL, and incubate at 37°C for 24 h. After incubation, carefully aspirate the bacterial solution, gently wash with PBS, and fix with anhydrous methanol for 15 min. After fixation, discard the methanol, dry, and stain with crystal violet for 15 min. After staining, remove the staining solution, wash with pure water, and dry thoroughly. Finally, decolorize with 33% acetic acid solution, place the 96-well plate in a multi-mode microplate reader, shake well, and measure the absorbance at 595 nm. Calculate the biofilm inhibition rate using the following formula: Biofilm inhibition rate = 1 - (Abs) 化合物 -Abs 空白对照 ) / (Abs 阴性对照 -Abs 空白对照 )×100%.

[0226] The test results are as follows Figure 2As shown, multiple target compounds effectively inhibited biofilm formation of Klebsiella pneumoniae KP31. Among them, compounds 1, 6, 7, 8, 9, 10, 11, and 32 exhibited significant biofilm inhibitory activity, with compounds 9, 10, and 11 showing the strongest inhibitory effects, achieving inhibition rates of 34%, 27%, and 23% on KP31 biofilm formation, respectively. Compared with the control group, compounds 2, 3, 4, 18, 22, 27, 28, 33, and 34 also showed certain biofilm inhibitory effects.

[0227] Experimental Example 4: Effect of Preferred Compounds on Cluster Motion of Klebsiella pneumoniae KP31

[0228] To further verify the quorum sensing inhibitory activity of the compounds of this invention, this experiment screened eight target compounds (compounds 1, 6, 7, 8, 9, 10, 11, and 13) with the best activity and systematically evaluated their intervention effect on the swarming ability of highly virulent multidrug-resistant Klebsiella pneumoniae KP31.

[0229] The specific experimental method is as follows: LB medium containing 0.3% (w / v) agar was prepared, autoclaved, and cooled to 60°C. The test compounds were added to final concentrations of 64 µg / mL, 128 µg / mL, and 256 µg / mL, respectively. After thorough mixing, the mixture was poured into 6 cm diameter petri dishes and allowed to solidify at room temperature. Subsequently, 1 μL of KP31 bacterial suspension was inoculated at the center of each plate and incubated at 37°C for 16 h. After incubation, bacterial growth was observed, bacterial plaque diameters (mm) were measured and photographed. By comparing the size and morphology of the bacterial plaques in each group, the effect of the compounds on the ability of bacteria to aggregate and move was evaluated.

[0230] Experimental results are as follows Figure 3 As shown, the untreated group of bacteria exhibited vigorous motility, with a diffusion patch diameter of 16.9 ± 0.8 mm. After intervention with the eight preferred compounds of this invention, the aggregation motility of the KP31 strain was significantly inhibited, with compound 1 showing the most prominent inhibitory effect. Moreover, the inhibitory effect of this compound showed a clear concentration dependence: at a low concentration of 32 µg / mL, the bacterial motility range was significantly limited, and the patch diameter was reduced to 11.5 ± 0.8 mm; when the concentration was increased to 64 µg / mL and 128 µg / mL, the patch diameter further decreased to 10.1 ± 0.3 mm and 9.7 ± 0.3 mm, respectively; and at the highest concentration of 256 µg / mL, the aggregation motility of KP31 was almost completely inhibited, with a patch diameter of only 7.9 ± 0.9 mm, demonstrating an extremely significant inhibitory effect.

[0231] Experimental Example 5: Effect of Preferred Compounds on Capsular Polysaccharide (CPS) of Klebsiella pneumoniae KP31

[0232] This experiment selected five target compounds (compounds 1, 6, 8, 11, and 13) with the best activity to investigate the effects of different concentration gradients of compounds on the capsular polysaccharide synthesis level of highly toxic multidrug-resistant Klebsiella pneumoniae KP31, and to further improve the evaluation system of the quorum sensing antiviral activity of the compounds of this invention.

[0233] The specific experimental method is as follows: KP31 cultures treated with 64, 128, and 256 µg / mL of the test compound for 48 h were collected. Anhydrous ethanol was added sequentially to a final concentration of 45%, and the mixture was centrifuged at 4 °C and 10,000 rpm for 10 min, retaining the supernatant. Anhydrous ethanol was then added to the supernatant to a final concentration of 90%, and the mixture was centrifuged at 4 °C and 10,000 rpm for 10 min, discarding the supernatant. The resulting precipitate was washed with acetone, dried overnight at room temperature, and dissolved in 1 mL of distilled water to obtain the test sample containing capsular polysaccharide.

[0234] Take 0.2 mL of the sample solution to be tested, add 1.2 mL of sodium tetraborate-sulfuric acid solution, heat in a 100 °C water bath for 5 min, cool in an ice-water bath, add 20 μL of 0.15% 3-phenylphenol reagent, mix gently, and measure the absorbance at 520 nm using a spectrophotometer within 5 min. Calculate the concentration of capsular polysaccharides in the sample according to the linear regression equation of the standard curve, and further calculate the inhibition rate of each compound on the production of capsular polysaccharides.

[0235] Experimental results are as follows Figure 4 As shown, compared with the blank control group, compounds 1, 8, 11, and 13 all inhibited capsular polysaccharide synthesis of Klebsiella pneumoniae KP31 in a dose-dependent manner, with significant differences in effective concentration and inhibitory activity among the different compounds. Compound 1 exhibited significant inhibitory effects at low concentrations, with CPS inhibition rates of 11%, 28%, and 32% at concentrations of 64, 128, and 256 μg / mL, respectively, all showing statistically significant differences compared to the blank control group. Compounds 8, 13, and 11 showed potent inhibitory activity at a high concentration of 256 μg / mL, with corresponding CPS inhibition rates of 41%, 42%, and 52%, respectively. Compound 6 did not show significant CPS inhibitory effects within the experimental concentration range and was not statistically significant compared to the control group.

[0236] Experimental Example 6: Synergistic effect of Compound 1 and antibacterial agent against Klebsiella pneumoniae KP31

[0237] Experimental method: Dilute the overnight culture of KP31 to OD200. 600≈ 0.05, then dispensed into test tubes. Experimental setups included a combination group of compound 1 with an antibiotic and a single-drug group: In the combination group, each tube contained compound 1 at a final concentration of 32 μg / mL, along with kanamycin (Kan, 0.4 μg / mL), streptomycin (Str, 0.5 μg / mL), azithromycin (Azm, 1.28 μg / mL), and chloramphenicol (Cm, 0.5 μg / mL); in the single-drug group, only the corresponding concentrations of the aforementioned antibiotics were added, without compound 1. After thorough mixing, the mixture was incubated at 37°C for 16 h, and the OD was measured. 600 Assess bacterial growth and calculate mortality rates for each treatment group accordingly.

[0238] Experimental results are as follows Figure 5 As shown, compound 1 exhibited significant synergistic bactericidal activity when used in combination with Kan, SPM, AZM, and CPL. When Kan, SPM, AZM, and CPL were used alone, the bacterial mortality rates were 9%, 11%, 13%, and 18%, respectively. When these antibiotics were combined with compound 1 to treat the bacterial strains, the bacterial mortality rates in all groups increased significantly. The bacterial mortality rates in the groups treated with compound 1 in combination with Kan, SPM, AZM, and CPL reached 44%, 42%, 44%, and 46%, respectively, representing increases of 35%, 31%, 31%, and 28% compared to the single-antibiotic treatment groups. These experimental results confirm that compound 1 can exert a synergistic sensitizing effect on multiple different types of antibiotics and can significantly improve the bactericidal efficiency of antibiotics against Klebsiella pneumoniae.

[0239] In summary, the series of functional verification experiments demonstrate that the compounds of this invention can effectively inhibit the synthesis of AI-2 quorum sensing signaling molecules in Klebsiella pneumoniae KP31, significantly weaken bacterial biofilm formation ability, inhibit bacterial swarming, and downregulate the expression levels of key virulence factors such as capsular polysaccharides, exhibiting excellent antiviral activity and quorum sensing inhibitory activity. Simultaneously, these compounds can broadly synergistically enhance the bactericidal effects of multiple classic antibiotics, significantly improving the killing effect of antibiotics on multidrug-resistant bacteria, and possessing excellent antibacterial sensitization capabilities. This has significant theoretical value and application prospects for solving the clinical challenges of multidrug-resistant bacterial infections and developing novel anti-infective drugs for the post-antibiotic era.

Claims

1. A benzenesulfonamide-based aryl amide derivative or a pharmaceutically acceptable salt or hydrate thereof, characterized in that, The derivative has a structure as shown in general formula I or II: ； Wherein, R is selected from C1-C8 straight-chain or branched alkyl, C3-C6 cycloalkyl, phenyl with different substituents, aryl with different substituents, C3-C6 arylcyclomethyl, and adamantyl. R1 is selected from hydrogen, C1-C8 straight-chain or branched alkyl groups, halogens, and C1-C8 straight-chain or branched alkoxy groups. R2 and R3 may be the same or different, and are independently selected from hydrogen, C1-C8 straight-chain or branched alkyl groups.

2. The benzenesulfonamide-based aryl amide derivative or its pharmaceutically acceptable salt or hydrate according to claim 1, characterized in that, R is selected from C3-C6 straight-chain or branched alkyl, C3-C6 cycloalkyl, phenyl with alkyl or trifluoromethyl substituents, aryl with alkoxycarbonyl substituents, C3-C6 arylcyclomethyl, and adamantyl. R1 is selected from hydrogen, C1-C8 straight-chain alkyl, halogen, and C1-C8 straight-chain alkoxy. R2 and R3 may be the same or different, and are independently selected from straight-chain alkyl groups of C1-C8.

3. The benzenesulfonamide-based aryl amide derivative or its pharmaceutically acceptable salt or hydrate according to claim 2, characterized in that, R1 is selected from methoxy, fluorine, and methyl; R2 is methyl; R3 is methyl.

4. The benzenesulfonamide-based aryl amide derivative or its pharmaceutically acceptable salt or hydrate according to claim 3, characterized in that, The R is selected from isopropyl, cyclohexyl, 4-ethylphenyl, 4-trifluoromethylphenyl, 3-trifluoromethylphenyl, 2-methylphenyl, 2-ethylphenyl, 4-ethoxycarbonylphenyl, 4-diethylaminophenyl, 5-ethoxycarbonyl-2-thiazolyl, 4-methoxycarbonyl-2-thiazolyl, 2-furanmethyl, and adamantyl. R1 is selected from methoxy, fluorine, and methyl; R2 is methyl; R3 is methyl.

5. The benzenesulfonamide-based aryl amide derivative according to claim 4, or its pharmaceutically acceptable salt or hydrate, characterized in that, The benzenesulfonamide-arylamide derivatives are selected from any of the following compounds:

6. A pharmaceutical composition, characterized in that, It comprises the benzenesulfonamide-aryl amide derivatives of any one of claims 1-5 or their pharmaceutically acceptable salts or hydrates, and a pharmaceutically acceptable carrier or excipient.

7. The pharmaceutical composition according to claim 6, characterized in that, It also includes at least one antibacterial active ingredient, which is kanamycin, streptomycin, azithromycin, chloramphenicol, ciprofloxacin, clarithromycin, tobramycin, norfloxacin, or levofloxacin.

8. The use of the benzenesulfonamide aryl amide derivative of any one of claims 1-5 or its pharmaceutically acceptable salt or hydrate, or the pharmaceutical composition of claim 6 or 7, in the preparation of bacterial quorum sensing inhibitors or antimicrobial sensitizers.

9. The application according to claim 8, characterized in that, The bacteria include Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, Salmonella, Shigella, Salmonella typhi, and Proteus.

10. The use of the benzenesulfonamide aryl amide derivative of any one of claims 1-5 or a pharmaceutically acceptable salt or hydrate thereof, or the pharmaceutical composition of claim 6 or 7, in the preparation of a medicament for the prevention and / or treatment of diseases caused by bacterial infections, including pneumonia, urinary tract infections, cystitis, diarrhea, sepsis, cholecystitis, endocarditis, and gastroenteritis.

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