1, 2, 4-triazole-Schiff base urease inhibitor as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medicines, in particular to a 1, 2, 4-triazole-Schiff base urease inhibitor as well as a preparation method and application thereof. The invention discloses a 1, 2, 4-triazole-Schiff base urease inhibitor for the first time, and the 1, 2, 4-triazole-Schiff base urease inhibitor has a good urease inhibition effect, can be used as a novel lead compound for helicobacter pylori resistance research, and is relatively low in toxicity to human normal cells and relatively high in safety. Meanwhile, the 1, 2, 4-triazole-Schiff base compound disclosed by the invention is simple in preparation method, relatively short in synthetic route, easy to prepare on a large scale and low in price.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, in particular to a 1,2,4-triazole-Schiff base urease inhibitor and a preparation method and application thereof. Background Art

[0002] Helicobacter pylori is considered the primary pathogen of chronic gastritis and gastric ulcers. It colonizes the gastric mucosa, triggering mucosal lesions, tissue damage, and an inflammatory response. The pathogenicity of this bacterium is centered on its production of urease, a recognized virulence factor. Urease is a nickel-containing polymerase that rapidly hydrolyzes urea into ammonium and carbon dioxide. This process alkalinizes the local pH environment, promoting bacterial colonization in the stomach and leading to the development of gastritis, gastric ulcers, duodenal ulcers, and even gastric cancer. Furthermore, urease has been associated with conditions such as urinary tract stones, pyelonephritis, hepatic encephalopathy, hepatic coma, urinary tract stones, and catheter obstruction. Despite advances in therapeutic interventions for these conditions, the clinical treatment of gastrointestinal ulcers remains challenging due to emerging antibiotic resistance and the inherent toxicity or side effects of existing drugs.

[0003] Growing evidence indicates that 1,2,4-triazole derivatives possess diverse biological activities, including antibacterial, insecticidal, antifungal, antitumor, antiproliferative, and antibiotic properties, and therefore hold great potential for pharmaceutical and agrochemical applications. Schiff bases, on the other hand, are organic molecules synthesized via amine-carbonyl condensation reactions and characterized by an azomethyl (-C=N-) group. Notably, these structural features appear to play a key role in both metal chelation and biological activity. In recent years, several 1,2,4-triazole or Schiff base derivatives have been shown to possess potent urease inhibitory activity. This discovery has greatly stimulated interest in the synthesis of novel 1,2,4-triazole-Schiff base analogs.

[0004] Therefore, how to develop a new drug that can effectively and safely treat gastric ulcer disease is still a technical problem that needs to be solved urgently by those skilled in the art. Summary of the Invention

[0005] In order to solve the above technical problems existing in the prior art, the present invention provides a 1,2,4-triazole-Schiff base urease inhibitor and a preparation method and application thereof.

[0006] A 1,2,4-triazole-Schiff base urease inhibitor, whose specific structural formula is as follows:

[0007]

[0008] R in the general formula I is at least one of 4-chloro, 4-bromo, 4-hydroxy, 2-hydroxy, 4-biphenyl, 3-fluoro, 4-methoxy, 4-fluoro, 2-fluoro, 2-chloro, 2,4-dihydroxy, 2-hydroxy, 5-bromo, 4-methyl, 2-hydroxy, 5-methyl, 2-hydroxy, 5-bromo, 2-hydroxy, 5-nitro, 2-nitro, 5-hydroxy, 2-hydroxy, and 3,5-dichloro.

[0009] Preferably, the specific structural formula of the 1,2,4-triazole-Schiff base urease inhibitor is as follows:

[0010]

[0011] The preparation method of the 1,2,4-triazole-Schiff base urease inhibitor comprises the following specific steps:

[0012] Step 1: A mixture of phenol and 32% sodium hydroxide solution is heated to 85°C; simultaneously, chloroacetic acid is slowly added dropwise, followed by continuous stirring at 85°C for 3 hours; after the reaction is complete, the mixture is poured into ice water, and the pH value of the solution is adjusted to 2-3 with hydrochloric acid; finally, the intermediate 2-phenoxyacetic acid is obtained by filtration and drying.

[0013]

[0014] Step 2: Reflux a mixture of 2-phenoxyacetic acid and thiocarbohydrazide at 160° C. for 1 hour. After the reaction is complete, crush the solid product, dilute with sufficient water, filter, and dry to obtain the intermediate 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol.

[0015]

[0016] Step 3: The intermediate 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol obtained above is mixed with differently substituted benzaldehydes in acetic acid and stirred at 120°C; after the reaction is completed, the mixture is cooled to room temperature until a large amount of solid precipitate is generated, and then the precipitate is filtered, washed with sufficient water and dried to obtain the target compound I, i.e., the 1,2,4-triazole-Schiff base urease inhibitor.

[0017]

[0018] Furthermore, in step 1, the ratio of phenol, chloroacetic acid and 32% sodium hydroxide is 25 mmol:45 mmol:8 mL.

[0019] Furthermore, in step 2, the ratio of 2-phenoxyacetic acid to thiocarbohydrazide is 1 mmol:1 mmol.

[0020] Furthermore, the ratio of the 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol, the different substituted benzaldehydes and the acetic acid is 1 mmol:1 mmol:10 mL.

[0021] Application of the 1,2,4-triazole-Schiff base urease inhibitor in the preparation of medicines.

[0022] Furthermore, the drug is an anti-Helicobacter pylori drug.

[0023] Furthermore, the drug is a gastric ulcer drug.

[0024] Furthermore, the drug is a drug that inhibits urease.

[0025] Compared with the prior art, the technical effects created by the present invention are embodied in:

[0026] 1. The present invention discloses for the first time a 1,2,4-triazole-Schiff base urease inhibitor, the specific structure of which is as follows:

[0027]

[0028] R in the general formula I is one of 4-chloro, 4-bromo, 4-hydroxy, 2-hydroxy, 4-biphenyl, 3-fluoro, 4-methoxy, 4-fluoro, 2-fluoro, 2-chloro, 2,4-dihydroxy, 2-hydroxy, 5-bromo, 4-methyl, 2-hydroxy, 5-methyl, 2-hydroxy, 5-bromo, 2-hydroxy, 5-nitro, 2-nitro, 5-hydroxy, 2-hydroxy, and 3,5-dichloro.

[0029] 2. The 1,2,4-triazole-Schiff base urease inhibitor provided by the present invention has good urease inhibition effect and can be used as a lead compound for novel anti-Helicobacter pylori research.

[0030] 3. The preparation method of the 1,2,4-triazole-Schiff base compound disclosed in the present invention is simple, the synthetic route is short, it is easy to prepare in large quantities, and the price is low.

[0031] 4. The 1,2,4-triazole-Schiff base urease inhibitors provided by the present invention have low toxicity to normal human cells and high safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The present invention is a preparation route of the 1,2,4-triazole-Schiff base urease inhibitor.

[0033] Figure 2 Cytotoxicity evaluation of compound 6 on HEK-293 cells. (ac: indicates significant difference, P < 0.05)

[0034] Figure 3 is the H NMR spectrum of compound 6.

[0035] Figure 4 is the carbon NMR spectrum of compound 6.

[0036] Figure 5 This is the high-resolution mass spectrum of compound 6. DETAILED DESCRIPTION

[0037] The technical solution of the present invention is further defined below in conjunction with specific implementation methods, but the scope of protection required is not limited to the description.

[0038] Example 1:

[0039] Preparation of (E)-4-((4-chlorobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 1)

[0040] The structural formula of compound 1 is shown below:

[0041]

[0042] The specific preparation steps are as follows:

[0043] Step 1: Heat a mixture of phenol and 32% sodium hydroxide solution to 85°C. Simultaneously, slowly add chloroacetic acid dropwise and continue stirring at 85°C for 3 hours. After the reaction is complete, pour the mixture into 100 mL of ice water and adjust the pH to 2-3 with hydrochloric acid. Finally, filter and dry to obtain the intermediate 2-phenoxyacetic acid.

[0044] Step 2: Reflux a mixture of 2-phenoxyacetic acid and thiocarbohydrazide at 160°C for 1 hour. After the reaction is complete, the solid product is crushed, diluted with sufficient water, filtered, and dried to obtain the intermediate 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol.

[0045] Step 3: The intermediate 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol obtained above was mixed with 4-chlorobenzaldehyde in acetic acid and stirred at 120°C for 4 hours. After the reaction was complete, the mixture was cooled to room temperature until a large amount of solid precipitate was formed. The precipitate was then filtered, washed with plenty of water, and dried to obtain the target compound 1.

[0046] Compound 1 is a white solid with a yield of 60.58% and a melting point of 201.4–205.9°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 1 are shown below:

[0047] 1 H NMR(400MHz, DMSO-d6)δ:14.16(s,1H),10.05(s,1H),7.87(d,J=7.6Hz,2H), 7.60(d,J=7.6Hz,2H),7.29(t,J=7.2Hz,2H),7.06-6.95(m,3H),5.26(s,2H).

[0048] 13 C NMR (100MHz, DMSO-d6) δ: 162.15, 161.92, 157.62, 147.05, 137.31, 130.89, 130.14, 129.47, 129.19, 121.56, 115.09, 59.62.

[0049] ESI-HRMS calculated for [MH] - C16H12ClN4OS - :343.0414,found:343.0422.

[0050] The preparation methods of the following examples are similar to those of Example 1, and the ratio of raw materials used is the same as that of Example 1, except that the 4-chlorobenzaldehyde in Example 1 is replaced by other corresponding benzaldehydes.

[0051] Example 2:

[0052] Preparation of (E)-4-((4-bromobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 2)

[0053] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 4-bromobenzaldehyde.

[0054] The structural formula of compound 2 is shown below:

[0055]

[0056] Compound 2 is a white solid with a yield of 61.20% and a melting point of 207.0–208.1°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 2 are shown below:

[0057] 1H NMR (400MHz, DMSO-d6) δ: 14.09 (s, 1H), 10.06 (s, 1H), 7.77 (d, J = 8.4Hz, 2H), 7.71(d,J=8.4Hz,2H)7.29(t,J=7.6Hz,2H),7.06-6.95(m,3H),5.26(s,2H).

[0058] 13 C NMR (100MHz, DMSO-d6) δ: 162.40, 162.11, 157.70, 147.18, 132.33, 131.32, 130.49, 129.67, 126.54, 121.69, 115.12, 59.61.

[0059] ESI-HRMS calculated for [MH] - C16H12BrN4OS - :386.9909,found:386.9919.

[0060] Example 3:

[0061] Preparation of (E)-4-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)phenol (Compound 3)

[0062] The difference compared with Example 1 is that only 4-chlorobenzaldehyde is replaced by 4-hydroxybenzaldehyde.

[0063] The structural formula of compound 3 is shown below:

[0064]

[0065] Compound 3 is a white solid with a yield of 57.10% and a melting point of 232.0–234.4°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 3 are shown below:

[0066] 1 H NMR(400MHz,DMSO-d6)δ:14.06(s,1H),10.39(s,1H),9.59(s,1H),7.70(d,J=7.2Hz ,2H),7.28(t,J=8.8Hz,2H),7.05-6.94(m,3H),6.89(d,J=7.6Hz,2H),5.21(s,2H).

[0067] 13C NMR (100MHz, DMSO-d6) δ: 165.19, 162.14, 161.97, 157.70, 146.90, 131.10, 129.68, 122.86, 121.67, 116.11, 115.11, 59.61.

[0068] ESI-HRMS calculated for [MH] - C 16 H 13 N4O2S - :325.0753,found:325.0763.

[0069] Example 4:

[0070] Preparation of (E)-2-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)phenol (Compound 4)

[0071] The difference compared with Example 1 is that only 4-chlorobenzaldehyde is replaced by 2-hydroxybenzaldehyde.

[0072] The structural formula of compound 4 is shown below:

[0073]

[0074] Compound 4 is a yellow solid with a yield of 36.92% and a melting point of 163.7–167.5°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 4 are shown below:

[0075] 1 H NMR (400MHz, DMSO-d6) δ: 14.10 (s, 1H), 10.45 (s, 1H), 10.16 (s, 1H), 7.77 (d, J = 7.6Hz, 1H), 7.41 (t, J = 8.0Hz, 1H), 7.29 (t, J = 7.2Hz, 2H), 7.04 (d, J = 7.6Hz, 2H), 6.98 (t, J = 7.2Hz, 2H), 6.89 (t, J = 7.6Hz, 1H), 5.25 (s, 2H).

[0076] 13 C NMR (100MHz, DMSO-d6) δ: 162.08, 160.78, 158.56, 157.66, 147.07, 134.49, 129.67, 127.48, 121.67, 119.68, 118.21, 116.72, 115.07, 59.63.

[0077] ESI-HRMS calculated for [MH] - C 16 H 13 N4O2S - :325.0753,found:325.0761.

[0078] Example 5:

[0079] Preparation of (E)-4-(([1,1'-biphenyl]-4-ylmethylene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 5)

[0080] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 4-biphenylbenzaldehyde.

[0081] The structural formula of compound 5 is shown below:

[0082]

[0083] Compound 5 is a white solid with a yield of 59.04% and a melting point of 156.0–158.9°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 5 are shown below:

[0084] 1 H NMR (400MHz, DMSO-d6) δ: 14.15 (s, 1H), 10.03 (s, 1H), 7.94 (d, J = 8.4Hz, 2H), 7.84 (d, J = 8.0Hz, 2H), 7.76 (d, J = 7.6Hz, 2H), 7. 50(t,J=7.6Hz,2H),7.42(t,J=7.2Hz,1H),7.30(t,J=7.6Hz,2H),7.05(d,J=8.4Hz,2H),6.98(t,J=7.2Hz,1H),5.28(s,2H).

[0085] 13 C NMR(100MHz,DMSO-d6)δ:163.15,162.17,157.63,147.00,144.09,138.88,130 .96,129.48,129.17,128.99,128.24,127.21,126.80,121.55,115.08,59.62.

[0086] ESI-HRMS calculated for [MH] - C 22 H 17 N4OS -:385.1117,found:385.1124.

[0087] Example 6:

[0088] Preparation of (E)-4-((3-fluorobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 6)

[0089] The difference compared with Example 1 is that only 4-chlorobenzaldehyde is replaced by 3-fluorobenzaldehyde.

[0090] The structural formula of compound 6 is shown below:

[0091]

[0092] Compound 6 is a white solid with a yield of 74.48% and a melting point of 172.5–175.9°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 6 are shown below:

[0093] 1 H NMR(400MHz,DMSO-d6)δ:14.18(s,1H),10.13(s,1H),7.71-7.63(m,2H),7.61-7.53(m,1H),7.4 5(t,J=8.8Hz,1H),7.33-7.25(m,2H),7.05(d,J=8.0Hz,2H),6.98(t,J=7.2Hz,1H),5.29(s,2H).

[0094] 13 C NMR (100MHz, DMSO-d6) δ: 163.61, 162.14, 161.72, 161.70 (d, 1C, J = 2.8Hz, 4 J CF ),161.17(d,1C,J=244.1Hz, 1 J CF ),157.75,147.33,134.58,134.50(d,1C,J=8.1Hz, 3 J CF ),131.49,131.41(d,1C,J=8.1Hz, 3 J CF ),129.67,125.50,121.71,119.79,119.58(d,1C,J=21.0Hz, 2 J CF ),115.15,114.43,114.20(d,1C,J=22.9Hz, 2J CF ),59.64.

[0095] ESI-HRMS calculated for [MH] - C 16 H 12 FN4OS - :327.0710,found:327.0719.

[0096] Example 7:

[0097] Preparation of (E)-4-((4-methoxybenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 7)

[0098] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 4-methoxybenzaldehyde.

[0099] The structural formula of compound 7 is shown below:

[0100]

[0101] Compound 7 is a white solid with a yield of 65.35% and a melting point of 150.4–152.5°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 7 are shown below:

[0102] 1 H NMR(400MHz, DMSO-d6)δ:13.99(s,1H),9.77(s,1H),7.81(d,J=8.8Hz,2H),7.28(t,J=6.8Hz,2H ),7.08(d,J=8.4Hz,2H),7.03(d,J=7.6Hz,2H),6.97(t,J=7.2Hz,1H),5.23(s,2H),3.85(s,3H).

[0103] 13 C NMR (100MHz, DMSO-d6) δ: 164.47, 163.03, 162.12, 157.69, 146.95, 130.80, 129.66, 124.43, 121.66, 115.08, 114.72, 59.60, 55.62.

[0104] ESI-HRMS calculated for [MH] - C 17 H 15 N4O2S - :339.0910,found:339.0919.

[0105] Example 8:

[0106] Preparation of (E)-4-((4-fluorobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 8)

[0107] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 4-fluorobenzaldehyde.

[0108] The structural formula of compound 8 is shown below:

[0109]

[0110] Compound 8 is a white solid with a yield of 47.24% and a melting point of 184.9–189.0°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 8 are shown below:

[0111] 1 H NMR (400MHz, DMSO-d6) δ: 14.08 (s, 1H), 9.99 (s, 1H), 7.92 (t, J = 6.0 Hz, 2H)), 7.36 (t, J = 8. 8Hz,2H),7.29(t,J=6.8Hz,2H),7.04(d,J=7.6Hz,2H),6.98(t,J=7.2Hz,1H),5.26(s,2H).

[0112] 13 C NMR (100MHz, DMSO-d6) δ: 166.06, 163.56 (d, 1C, J = 250.3Hz, 1 J CF ),162.97,162.13,157.71,147.13,131.39,131.30(d,1C,J=9.1Hz, 3 J CF ),129.69,128.72,128.69(d,1C,J=3.1Hz, 4 J CF ),121.71,116.60,116.38(d,1C,J=22.1Hz, 2 J CF ),115.13,59.63.

[0113] ESI-HRMS calculated for [MH] - C 16 H 12 FN4OS -:327.0710,found:327.0719.

[0114] Example 9:

[0115] Preparation of (E)-4-((2-fluorobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 9)

[0116] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 2-fluorobenzaldehyde.

[0117] The structural formula of compound 9 is shown below:

[0118]

[0119] Compound 9 is a white solid with a yield of 61.06% and a melting point of 172.6–179.9°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 9 are shown below:

[0120] 1 H NMR (400MHz, DMSO-d6) δ: 14.11 (s, 1H), 10.52 (s, 1H), 7.92 (t, J = 7.6Hz, 1H), 7.70-7.6 1(m,1H),7.43-7.25(m,4H),7.05(d,J=8.0Hz,2H),6.98(d,J=7.2Hz,1H),5.29(s,2H).

[0121] 13 C NMR (100MHz, DMSO-d6) δ: 163.09, 162.00, 160.56 (d, 1C, J = 252.6Hz, 1 J CF ),157.76,154.98,154.92(d,1C,J=5.1Hz, 3 J CF )147.44,135.13,135.04(d,1C,J=8.8Hz, 3 J CF ),129.67,127.28,125.35,125.32(d,1C,J=3.2Hz, 4 J CF ),121.67,119.93,119.83(d,1C,J=9.7Hz, 3 J CF ),116.61,116.41(d,1C,J=20.5Hz, 2 J CF),115.13,59.61.

[0122] ESI-HRMS calculated for [MH] - C 16 H 12 FN4OS - :327.0710,found:327.0720.

[0123] Example 10:

[0124] Preparation of (E)-4-((2-chlorobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 10)

[0125] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 2-chlorobenzaldehyde.

[0126] The structural formula of compound 10 is shown below:

[0127]

[0128] Compound 10 is a white solid with a yield of 60.42% and a melting point of 158.4–160.5°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 10 are shown below:

[0129] 1 H NMR (400MHz, DMSO-d6) δ: 14.14 (s, 1H), 10.82 (s, 1H), 7.99 (d, J = 8.4Hz, 1H), 7.65-7.56 (m, 2H), 7.4 3(t,J=8.0Hz,1H),7.30(t,J=8.0Hz,2H),7.06(d,J=8.0Hz,2H),6.99(t,J=7.2Hz,1H),5.31(s,2H).

[0130] 13 C NMR (100MHz, DMSO-d6) δ: 161.98, 157.79, 156.79, 147.60, 135.21, 134.14, 130.36, 129.87, 129.68, 127.96, 127.69, 121.67, 115.14, 59.63.

[0131] ESI-HRMS calculated for [MH] - C 16 H 12 ClN4OS -:343.0414,found:343.0424.

[0132] Example 11:

[0133] Preparation of (E)-4-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)benzene-1,3-diol (Compound 11)

[0134] The difference compared with Example 1 is that only 4-chlorobenzaldehyde is replaced by 2,4-dihydroxybenzaldehyde.

[0135] The structural formula of compound 11 is shown below:

[0136]

[0137] Compound 11 is a light yellow solid with a yield of 49.34% and a melting point of 237.2–240.7°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 11 are shown below:

[0138] 1 H NMR(400MHz, DMSO-d6)δ:14.02(s,1H),10.40(s,1H),10.30(s,1H),9.71(s,1H),7.61(d,J=8.4Hz,1H),7 .29(dd,J=8.8,7.2Hz,2H),7.02(d,J=8.4Hz,2H),6.97(t,J=7.2Hz,1H),6.37-6.32(m,2H),5.20(s,2H).

[0139] 13 C NMR (100MHz, DMSO-d6) δ: 163.52, 162.90, 162.22, 160.75, 157.62, 146.76, 129.98, 129.69, 121.68, 115.08, 109.76, 108.70, 102.45, 59.64.

[0140] ESI-HRMS calculated for [MH] - C 16 H 13 N4O3S - :341.0702,found:341.0709.

[0141] Example 12:

[0142] Preparation of (E)-4-bromo-2-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)phenol (Compound 12)

[0143] The difference compared with Example 1 is that only 4-chlorobenzaldehyde is replaced by 2-hydroxy-5-bromobenzaldehyde.

[0144] The structural formula of compound 12 is shown below:

[0145]

[0146] Compound 12 is a light yellow solid with a yield of 59.94% and a melting point of 200.5–202.2°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 12 are shown below:

[0147] 1 H NMR(400MHz, DMSO-d6)δ:14.12(s,1H),10.79(s,1H),10.30(s,1H),7.86(s,1H),7.55(dt,J= 8.8, 2.8Hz, 1H), 7.30 (t, J = 8.0Hz, 2H), 7.05 (d, J = 7.6Hz, 2H), 7.02-6.90 (m, 2H), 5.28 (s, 2H).

[0148] 13 C NMR (100MHz, DMSO-d6) δ: 162.02, 157.74, 157.68, 157.53, 147.34, 136.57, 129.67, 128.73, 121.74, 120.57, 119.04, 115.17, 110.95, 59.74.

[0149] ESI-HRMS calculated for [MH] - C 16 H 12 BrN4O2S - :402.9858,found:402.9870.

[0150] Example 13:

[0151] Preparation of (E)-4-((4-methylbenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol (Compound 13)

[0152] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 4-methylbenzaldehyde.

[0153] The structural formula of compound 13 is shown below:

[0154]

[0155] Compound 13 is a white solid with a yield of 65.98% and a melting point of 175.8–179.4°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 13 are shown below:

[0156] 1 H NMR(400MHz, DMSO-d6)δ:14.14(s,1H),9.87(s,1H),7.74(d,J=8.0Hz,2H),7.33(d,J=8.0Hz,2H ),7.28(t,J=8.0Hz,2H),7.03(d,J=8.4Hz,2H),6.97(t,J=7.2Hz,1H),5.24(s,2H),2.37(s,3H).

[0157] 13 C NMR (100MHz, DMSO-d6) δ: 164.15, 162.11, 157.69, 147.05, 143.25, 129.80, 129.64, 129.34, 128.75, 121.65, 115.08, 59.60, 21.33.

[0158] ESI-HRMS calculated for [MH] - C 17 H 15 N4OS - :323.0961,found:323.0970.

[0159] Example 14:

[0160] Preparation of (E)-2-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)-4-methylphenol (Compound 14)

[0161] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 2-hydroxy-5-methylbenzaldehyde.

[0162] The structural formula of compound 14 is shown below:

[0163]

[0164] Compound 14 is a light yellow solid with a yield of 62.04% and a melting point of 175.3–178.5°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 14 are shown below:

[0165] 1 H NMR(400MHz, DMSO-d6)δ:13.99(s,1H),10.11(s,1H),10.10(s,1H),7.53(s,1H),7.29(t,J=8.0,2H),7.22(dd,J =8.4, 2.4Hz, 1H), 7.05 (d, J = 8.4Hz, 2H), 6.98 (t, J = 7.2Hz, 1H), 6.88 (d, J = 8.4Hz, 1H), 5.25 (s, 2H), 2.20 (s, 3H).

[0166] 13 C NMR(100MHz,DMSO-d6)δ:162.17,161.09,157.60,156.36,146.85,135.02 ,129.43,128.17,127.34,121.55,117.68,116.52,115.08,59.71,19.81.

[0167] ESI-HRMS calculated for [MH] - C 17 H 15 N4O2S - :339.0910,found:339.0915.

[0168] Example 15:

[0169] Preparation of (E)-4-chloro-2-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)phenol (Compound 15)

[0170] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 2-hydroxy-5-chlorobenzaldehyde.

[0171] The structural formula of compound 15 is shown below:

[0172]

[0173] Compound 15 is a white solid with a yield of 61.80% and a melting point of 206.5–211.4°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 15 are shown below:

[0174] 1H NMR (400MHz, DMSO-d6) δ: 14.14 (s, 1H), 10.81 (s, 1H), 10.33 (s, 1H), 7.72 (d, J = 2.8Hz, 1H), 7.4 3(dd,J=8.8,2.8Hz,1H),7.32-7.26(m,2H),7.06-7.02(m,2H),7.01-6.95(m,2H),5.28(s,2H).

[0175] 13 C NMR (100MHz, DMSO-d6) δ: 162.07, 157.67, 157.63, 157.10, 147.15, 133.52, 129.45, 125.78, 123.36, 121.59, 119.91, 118.50, 115.10, 59.71.

[0176] ESI-HRMS calculated for [MH] - C 16 H 12 ClN4O2S - :359.0364,found:359.0371.

[0177] Example 16:

[0178] Preparation of (E)-2-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)-4-nitrophenol (Compound 16)

[0179] The difference compared with Example 1 is that only 4-chlorobenzaldehyde is replaced by 2-hydroxy-5-nitrobenzaldehyde.

[0180] The structural formula of compound 16 is shown below:

[0181]

[0182] Compound 16 is a yellow solid with a yield of 44.73% and a melting point of 213.8–216.9°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 16 are shown below:

[0183] 1H NMR(400MHz, DMSO-d6)δ:14.17(s,1H),12.10(s,1H),10.44(s,1H),8.63(d,J=3.2Hz,1H),8.28(dd,J=9.2,2.8 Hz,1H),7.32-7.25(m,2H),7.17(d,J=9.2Hz,1H),7.05(dd,J=8.8,0.8Hz,2H),6.99-6.93(m,1H),5.30(s,2H).

[0184] 13 C NMR (100MHz, DMSO-d6) δ: 163.62, 162.08, 157.69, 156.74, 147.36, 140.06, 129.65, 129.17, 122.66, 121.70, 119.04, 117.46, 115.20, 59.77.

[0185] ESI-HRMS calculated for [MH] - C 16 H 12 N5O4S - :370.0604,found:370.0612.

[0186] Example 17:

[0187] Preparation of (E)-3-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)-4-nitrophenol (Compound 17)

[0188] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 2-nitro-5-hydroxybenzaldehyde.

[0189] The structural formula of compound 17 is shown below:

[0190]

[0191] Compound 17 is a yellow solid with a yield of 47.53% and a melting point of 204.5–208.0°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 17 are shown below:

[0192] 1H NMR (400MHz, DMSO-d6) δ: 14.21 (s, 1H), 11.34 (s, 1H), 10.87 (s, 1H), 8.16 (dd, J = 9.2, 2.4Hz, 1H), 7.37 (s, 1H) ),7.32-7.26(m,2H),7.11(dt,J=9.2,2.8Hz,1H),7.04(d,J=6.8Hz,2H),6.97(t,J=7.2Hz,1H),5.28(s,2H).

[0193] 13 C NMR (100MHz, DMSO-d6) δ: 162.80, 162.31, 158.48, 157.59, 147.48, 140.09, 130.36, 129.70, 128.38, 121.75, 118.80, 115.11, 115.05, 59.49.

[0194] ESI-HRMS calculated for [MH] - C 16 H 12 N5O4S - :370.0604,found:370.0609.

[0195] Example 18:

[0196] Preparation of (E)-2,4-dichloro-6-(((3-mercapto-5-(phenoxymethyl)-4H-1,2,4-triazol-4-yl)imino)methyl)phenol (Compound 18)

[0197] Compared with Example 1, the difference is that only 4-chlorobenzaldehyde is replaced by 2-hydroxy-3,5-dichlorobenzaldehyde.

[0198] The structural formula of compound 18 is shown below:

[0199]

[0200] Compound 18 was a white solid with a yield of 64.86% and a melting point of 186.0–189.6°C. The H-NMR spectrum, C-NMR spectrum, and high-resolution mass spectrum data of compound 18 are shown below:

[0201] 1H NMR(400MHz,DMSO-d6)δ:14.20(s,1H),10.84(s,1H),10.42(s,1H),7.79(s,1H),7.73( s,1H),7.30(t,J=7.6Hz,2H),7.05(d,J=8.0Hz,2H),6.98(t,J=7.2Hz,1H),5.32(s,2H).

[0202] 13 C NMR (100MHz, DMSO-d6) δ: 162.28, 158.33, 157.52, 152.63, 147.01, 132.71, 129.44, 126.25, 123.92, 122.99, 121.74, 121.62, 115.10, 59.73.

[0203] ESI-HRMS calculated for [MH] - C 16 H 11 Cl2N4O2S - :392.9974,found:392.9986.

[0204] In order to further verify the excellent effect of the present invention, the inventors also conducted the following comparative experiments:

[0205] Experiment 1

[0206] First, add 25 μL of urease solution (0.017 mg / mL) and 25 μL of test compound to a 96-well plate, and then place the plate in a 37°C incubator for 30 minutes. Next, add 50 μL of urea solution containing 3.2 mmol / L (pH = 7.8, phosphate buffer) and incubate at 37°C for 30 minutes. Finally, add 50 μL of solution A (1% w / v phenol and 0.005% w / v sodium nitroprusside) and solution B (0.5% w / v NaOH and 0.1% w / v NaOCl) and incubate at 37°C for 20 minutes. After 20 minutes, measure the absorbance at a wavelength of 625 nm. Each sample was tested three times and the IC was calculated. 50 , the results are shown in Table 1.

[0207] Table 1. α-glucosidase inhibitory activity (IC) of coumarin-oxadiazole compounds 50 )

[0208]

[0209] al, indicates significant difference, P<0.05.

[0210] It can be seen from Table 1 that most of the compounds synthesized in the present invention have good urease inhibition activity, and compound 6 has the strongest urease activity, with an IC 50 It was 6.68±0.44μM, which was better than the positive control drug thiourea (41.56±1.26μM).

[0211] Experiment 2

[0212] HEK-293 cells (1.0×10 5 cells / mL) were seeded into 96-well plates and incubated at 37°C for 24 hours. The cells were then treated with various concentrations of compound 6 (10, 20, 40, 80, and 160 μM) and incubated for an additional 24 hours. 10 μl of MTT solution was added, and the plates were incubated for 4 hours. After removing the supernatant, 150 μl of DMSO was added to dissolve the crystals, and the absorbance at 490 nm was measured.

[0213] The results are as follows Figure 2 As shown in Figure 3, when the concentration of compound 6 was in the range of 10-160 μM, the cell viability was always maintained above 95%. 50 =6.68±0.44 μM) showed negligible cytotoxicity against HEK-293 cells at low concentrations. In conclusion, compound 6 is a promising candidate for anti-Helicobacter pylori therapy with a high safety profile.

[0214] Finally, it should be noted that the above embodiments are merely representative examples of the present invention. Obviously, the technical solutions of the present invention are not limited to the above embodiments and are subject to numerous variations. All variations that can be directly derived or conceived by a person of ordinary skill in the art from the disclosure of the present invention should be considered within the scope of protection of the present invention.

Claims

1. A 1,2,4-triazole-Schiff base urease inhibitor, characterized in that: Its specific structural formula is as follows:

2. The 1,2,4-triazole-Schiff-base urease inhibitor according to claim 1, wherein R in the general formula I is at least one of 4-chloro, 4-bromo, 4-hydroxy, 2-hydroxy, 4-biphenyl, 3-fluoro, 4-methoxy, 4-fluoro, 2-fluoro, 2-chloro, 2,4-dihydroxy, 2-hydroxy, 5-bromo, 4-methyl, 2-hydroxy, 5-methyl, 2-hydroxy, 5-bromo, 2-hydroxy, 5-nitro, 2-nitro, 5-hydroxy, 2-hydroxy, and 3,5-dichloro.

3. The 1,2,4-triazole-Schiff-base urease inhibitor according to claim 1, characterized in that The specific structural formula of the 1,2,4-triazole-Schiff base urease inhibitor is as follows:

4. The method for preparing the 1,2,4-triazole-Schiff base urease inhibitor according to claim 1, characterized in that: The specific steps are as follows: Step 1: A mixture of phenol and 32% sodium hydroxide solution is heated to 85°C; simultaneously, chloroacetic acid is slowly added dropwise, followed by continuous stirring at 85°C for 3 hours; after the reaction is complete, the mixture is poured into ice water, and the pH value of the solution is adjusted to 2-3 with hydrochloric acid; finally, the intermediate 2-phenoxyacetic acid is obtained by filtration and drying. Step 2: Reflux the mixture of 2-phenoxyacetic acid and thiocarbohydrazide at 160° C. for 1 hour. After the reaction is complete, crush the solid product, dilute with sufficient water, filter, and dry to obtain the intermediate 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol. Step 3: The intermediate 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol obtained above is mixed with differently substituted benzaldehydes in acetic acid and stirred at 120°C; after the reaction is completed, the mixture is cooled to room temperature until a large amount of solid precipitate is generated, and then the precipitate is filtered, washed with sufficient water and dried to obtain the target compound I, i.e., the 1,2,4-triazole-Schiff base urease inhibitor.

5. The method for preparing a 1,2,4-triazole-Schiff base urease inhibitor according to claim 4, wherein: In the step 1, the ratio of phenol, chloroacetic acid and 32% sodium hydroxide is 25 mmol:45 mmol:8 mL.

6. The method for preparing a 1,2,4-triazole-Schiff base urease inhibitor according to claim 4, wherein: In the step 2, the ratio of 2-phenoxyacetic acid to thiocarbohydrazide is 1 mmol:1 mmol.

7. The method for preparing a 1,2,4-triazole-Schiff base urease inhibitor according to claim 4, wherein: In step 3, the ratio of 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol, different substituted benzaldehydes and acetic acid is 1 mmol:1 mmol:10 mL.

8. Use of the 1,2,4-triazole-Schiff-base urease inhibitor according to claim 1 in the preparation of medicines.

9. Use of the 1,2,4-triazole-Schiff-base urease inhibitor according to claim 8 in the preparation of medicines, characterized in that: The drug is an anti-Helicobacter pylori drug.

10. Use of the 1,2,4-triazole-Schiff-base urease inhibitor according to claim 8 in the preparation of a drug for treating liver fibrosis, characterized in that: The medicine is a gastric ulcer medicine.

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