1,2,4-triazole-coumarin-based dual alpha-glucosidase and ptp1b inhibitors, methods of preparation and uses

Abstract

This invention discloses a 1,2,4-triazole-coumarin dual-target inhibitor of α-glucosidase and protein tyrosine phosphatase 1B (PTP1B), its preparation method, and its applications. The chemical structure of this dual-target inhibitor of α-glucosidase and PTP1B is shown in formula (I). The 1,2,4-triazole-coumarin compounds disclosed in this invention have significant inhibitory effects on the activity of α-glucosidase and PTP1B and can reduce postprandial blood glucose levels, making them potential lead compounds for novel antidiabetic research. The preparation conditions of the 1,2,4-triazole-coumarin compounds disclosed in this invention are mild, the operation is simple, the price is low, and it is easy to promote.

Description

Technical Field

[0001] This invention belongs to the field of drug synthesis technology, specifically relating to a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B, its preparation method, and its application in the preparation of antidiabetic drugs. Background Technology

[0002] Diabetes mellitus (DM) is a chronic metabolic disease characterized by long-term hyperglycemia, caused by defects in insulin secretion or action. Hyperglycemia can damage the kidneys, eyes, blood vessels, and nerves, leading to diabetic nephropathy, retinopathy, cardiovascular and cerebrovascular diseases, and neuropathy. It severely impacts patients' quality of life and has become one of the leading causes of death worldwide, placing a heavy burden on society and families.

[0003] In the pathological process of diabetes, alpha-glucosidase plays a crucial role. It resides in the epithelial cells of the small intestinal villi and is responsible for breaking down carbohydrates into glucose. Excessive activity of alpha-glucosidase can cause a sharp rise in postprandial blood glucose in diabetic patients. Currently, while alpha-glucosidase inhibitors such as miglitol, acarbose, and voglibose can inhibit its activity and lower postprandial blood glucose, they have side effects such as gastrointestinal discomfort, and their efficacy is poor in some patients. Therefore, the development of better drugs is essential.

[0004] Meanwhile, protein tyrosine phosphatase 1B (PTP1B) negatively regulates the insulin signaling pathway by dephosphorylating tyrosine residues of insulin receptors or substrates, leading to insulin resistance, a crucial pathogenesis of type 2 diabetes. Although researchers have discovered numerous natural and synthetic compounds with PTP1B inhibitory activity, none have yet been approved for clinical use due to low bioavailability, metabolic instability, and significant toxic side effects. Given the current lack of highly effective and well-tolerated diabetes treatments in clinical practice, researchers need to conduct in-depth research into the pathogenesis and develop novel hypoglycemic drugs using advanced technologies. Summary of the Invention

[0005] In view of this, the object of the present invention is to provide a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B, its preparation method, and its application in the preparation of antidiabetic drugs. This inhibitor can be used to develop a highly effective, safe, and well-tolerated diabetes treatment drug, which is a novel hypoglycemic agent.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] The first technical objective of this invention is to provide a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B, having the structure shown in formula (I): ; In general formula I, R is at least one of 3-hydroxy, 4-hydroxy, 4-hydroxy-3,5-dimethyl, 4-methyl, 3-chloro-4-hydroxy, 4-fluoro, 2-fluoro-4-hydroxy, 4-hydroxy-2,6-dimethyl, 3-nitro-4-hydroxy, 4-chloro, 4-bromo, 3-bromo-4-hydroxy-5-nitro, 3-chloro, and 4-biphenyl.

[0008] It should be noted that, in this invention, triazole-containing heterocyclic compounds, due to their unique chemical structures, exhibit broad pharmacological activities. In the treatment of diabetes, they can inhibit α-glucosidase and affect the insulin signaling pathway. They also show significant activity in anticancer and antiviral fields, providing new insights for diabetes treatment. Similarly, coumarin derivatives have been synthesized in large quantities as α-glucosidase inhibitors, which are not only highly active and selective with low side effects, but also inhibit PTP1B, showing promise as dual-target antidiabetic drugs.

[0009] Exemplary, the structural formula of the dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B includes:

[0010]

[0011] .

[0012] The second technical objective of this invention is to request protection of a method for preparing a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B as described above. The synthesis of this type of compound uses 4-hydroxycoumarin as the starting material. The entire synthetic route is ingeniously designed, with not only simplified steps, but also mild reaction conditions at each step, making it easy to operate and possessing good prospects for industrial application.

[0013] Furthermore, the method specifically includes the following steps: Step 1: Add 4-hydroxycoumarin and epichlorohydrin solution to sodium hydroxide, then add anhydrous ethanol as solvent, and reflux at 80°C for 4-10 hours; after the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain the product 4-(epoxyethylene-2-ylmethoxy)-2H-benzopyran-2-one.

[0014] Step 2: Phenol and chloroacetic acid are reacted in an ethanol solution of sodium hydroxide at 80°C for 4-10 hours. After the reaction is completed, hydrochloric acid is added to adjust the pH to 2-3. The mixture is allowed to stand at 4°C to crystallize and then filtered to obtain the product 2-phenoxyacetic acid.

[0015] Step 3: Reflux the product 2-phenoxyacetic acid with aminothiourea at 160℃ for 1-3 h, wash the reaction mixture with H2O, filter and dry to obtain the filter residue, which is the product 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol.

[0016] Step 4: The product 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol is reacted with benzaldehyde with different substituents in glacial acetic acid solvent at 120℃ for 2-8 hours. After standing, the precipitate is collected by filtration to obtain the residue, which is (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol with different substituents.

[0017] Step 5: The obtained (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol, 4-(ethylene oxide-2-ylmethoxy)-2H-benzopyran-2-one, and anhydrous ethanol were refluxed at 80°C for 4-10 h. After the reaction was completed, the target compound I was obtained by column chromatography with petroleum ether:ethyl acetate = 3:1, i.e., the 1,2,4-triazol-coumarin α-glucosidase and PTP1B dual-target inhibitor.

[0018]

[0019] Preferably, the ratio of 4-hydroxycoumarin, epichlorohydrin, sodium hydroxide and anhydrous ethanol is 18 mmol: 235 mmol: 20 mmol: 150 mL.

[0020] Preferably, the ratio of phenol, chloroacetic acid, sodium hydroxide and anhydrous ethanol is 21 mmol: 38 mmol: 127 mmol: 15 mL.

[0021] Preferably, the molar ratio of 2-phenoxyacetic acid to aminothiourea is 1 mmol: 1 mmol.

[0022] Preferably, the ratio of benzaldehyde with different substituents, 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol and glacial acetic acid is 1 mmol:1.2 mmol:6 mL.

[0023] Preferably, the ratio of (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol, 4-(ethylene oxide-2-ylmethoxy)-2H-benzopyran-2-one and anhydrous ethanol is 1 mmol:1 mmol:35 mL.

[0024] The third technical objective of this invention is to request protection for the use of the above-mentioned 1,2,4-triazole-coumarin α-glucosidase and PTP1B dual-target inhibitors in the preparation of antidiabetic drugs.

[0025] Compared with the prior art, the present invention discloses a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B, its preparation method, and its application, which has the following advantages: 1. The 1,2,4-triazole-coumarin dual-target inhibitors of α-glucosidase and PTP1B disclosed in this invention have good inhibitory effects on the activity of α-glucosidase and PTP1B and can reduce postprandial blood glucose. They can be used as lead compounds for novel anti-diabetic research. 2. The 1,2,4-triazole-coumarin compounds disclosed in this invention have mild preparation conditions, are simple to operate, are inexpensive, and are easy to prepare in large quantities. Attached Figure Description

[0026] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0027] Figure 1 This is a preparation route diagram for the dual-target inhibitors of 1,2,4-triazole-coumarin α-glucosidase and PTP1B.

[0028] Figure 2 The effect of compound 10n on postprandial blood glucose in Kunming mice was investigated. (A) shows the blood glucose inhibition effect of compound 10n on Kunming mice after sucrose administration; (B) shows the AUC of the mice after sucrose administration. 0-150 min The increment, "" indicates a significant difference, P<0.05.

[0029] Figure 3 The image shows the proton NMR spectrum of compound 10h.

[0030] Figure 4 The image shows the carbon NMR spectrum of compound 10h.

[0031] Figure 5 This is a high-resolution mass spectrum of compound 10h.

[0032] Figure 6 The image shows the proton NMR spectrum of compound 10n.

[0033] Figure 7 This is the carbon NMR spectrum of compound 10n.

[0034] Figure 8 This is a high-resolution mass spectrum of compound 10n. Detailed Implementation

[0035] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] To better understand the present invention, the following embodiments are provided for further detailed description of the present invention, but they should not be construed as limiting the present invention. Any non-essential improvements and adjustments made by those skilled in the art based on the above-described invention are also considered to fall within the protection scope of the present invention.

[0037] This invention discloses a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B, and its preparation method.

[0038] To further illustrate the technical solution disclosed in this invention, the inventors have also provided the following embodiments: Example 1: Preparation of (E)-4-(2-hydroxy-3-((4-((3-hydroxybenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)propoxy)-2H-benzopyran-2-one (compound 10a) The structural formula of compound 10a is shown below:

[0039] The specific preparation steps are as follows: Step 1: Add 4-hydroxycoumarin and epichlorohydrin solution to sodium hydroxide, then add anhydrous ethanol as solvent, and reflux at 80°C for 6 hours; after the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain the product 4-(epoxyethylene-2-ylmethoxy)-2H-benzopyran-2-one. Step 2: Phenol and chloroacetic acid were reacted in an ethanol solution of sodium hydroxide at 80°C for 6 hours. After the reaction was completed, hydrochloric acid was added to adjust the pH to 3, and the mixture was allowed to stand at 4°C to crystallize. The product 2-phenoxyacetic acid was obtained by filtration. Step 3: Reflux the product 2-phenoxyacetic acid and aminothiourea at 160℃ for 3 hours, take the mixture and H2O, filter and dry to obtain the filter residue, which is the product 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol. Step 4: The product 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol was reacted with 3-hydroxybenzaldehyde in glacial acetic acid solvent at 120°C for 6 hours. After standing, the precipitate was collected by filtration, and the residue was obtained, which is 3-hydroxy-substituted (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazole-3-thiol. Step 5: The 3-hydroxy-substituted (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol, 4-(ethylene oxide-2-ylmethoxy)-2H-benzopyran-2-one, and anhydrous ethanol obtained above were refluxed at 80 °C for 6 h; after the reaction was completed, the target compound 10a was obtained by column chromatography with petroleum ether:ethyl acetate = 3:1.

[0040] Compound 10a was a white solid with a yield of 47% and a melting point of 187.0–190.0 °C. The 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10a are shown below: 1 H NMR (DMSO, 400 MHz) d 3.47 (d, J = 5.2 Hz, 1H), 4.16-4.24 (m, 3H), 5.18 (s, 2H), 5.75 (d, J = 5.2 Hz, 1H), 5.84 (s, 1H), 6.83-6.85 (m, 2H), 6.90-6.94 (m, 1H), 6.97-6.99 (m, 2H), 7.22-7.37 (m, 4H), 7.60-7.65 (m, 3H), 7.87(dd, 1H, J = 8.0 Hz, 1.6 Hz), 8.69 (s, 1H), 10.45 (s, 1H); 13 C NMR (100 Hz, DMSO) d: 35.7, 60.0, 67.6, 72.5, 91.2, 115.5, 115.7, 116.6, 116.9, 122.1, 123.0, 123.8, 124.7, 130.1, 131.9, 133.3, 148.6, 149.1, 153.3, 157.9, 162.2, 162.8, 165.5, 167.3; HRMS (ESI) calcd for [M+H] + C 28 H 25 N4O6S + : 545.1489 found 545.1486. The preparation methods of the following examples are similar to those of Example 1, and the proportions of the raw materials used are the same as those of Example 1. The only difference is that the 3-hydroxybenzaldehyde in Example 1 is replaced with other corresponding substituted benzaldehydes.

[0041] Example 2: Preparation of (E)-4-(2-hydroxy-3-((4-((4-hydroxybenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)propoxy)-2H-benzopyran-2-one (compound 10b) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 4-hydroxybenzaldehyde.

[0042] The structural formula of compound 10b is shown below:

[0043] Compound 10b is a white powder with a yield of 55% and a melting point of 178.6-181.3 °C. The 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10b are shown below: 1 H NMR (DMSO, 400 MHz) d 3.48 (d, J = 8.4 Hz, 1H), 4.16-4.24 (m, 3H), 5.18 (s, 2H), 5.75 (d, J = 5.2 Hz, 1H), 5.84 (s, 1H), 6.82-6.85 (m, 2H), 6.90-6.99 (m, 3H), 7.22-7.37 (m, 4H), 7.60-7.66 (m, 3H), 7.88 (d, J= 7.6 Hz, 1H), 8.69 (s, 1H), 10.45 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 36.0, 60.2, 67.8, 72.5, 91.2, 115.6, 115.7, 115.8, 116.6, 116.71, 122.0, 123.1, 123.8, 124.6, 130.0, 131.8, 133.2, 148.7, 149.0, 153.3, 158.1, 162.1, 162.8, 165.5, 167.2, 167.2; HRMS (ESI) calcd for [M+H] + C 28 H 25 N4O6S + : 545.1489 found 545.1489. Example 3: Preparation of (E)-4-(2-hydroxy-3-((4-((4-hydroxy-3,5-dimethylbenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)propoxy)-2H-benzopyran-2-one (compound 10c) Compared to Example 1, the only difference is that the 3-hydroxyl group is replaced with 4-hydroxy-3,5-dimethyl.

[0044] The structural formula of compound 10c is shown below:

[0045] Compound 10c is a pink powder with a yield of 51% and a melting point of 213.2-215.4 °C. The 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10c are shown below: 1 H NMR (DMSO, 400 MHz) d : 2.12 (s, 6H), 3.46 (d, J = 8.8 Hz, 1H), 4.16-4.23 (m, 3H), 5.17 (s, 2H), 5.73 (d, J = 4.8 Hz, 1H), 5.82 (s, 1H), 6.90-6.99(m, 3H), 7.22-7.36 (m, 6H), 7.60-7.63 (m, 1H), 7.86 (d,J = 8.0 Hz, 1H), 8.60(s, 1H), 9.26 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 17.0, 17.1, 35.9, 60.0, 67.6, 72.5, 91.2, 115.4, 115.5, 115.7, 116.9, 122.1, 122.8, 123.8, 124.6, 125.5, 129.6, 129.8, 130.1, 130.3, 133.3, 148.7, 149.0, 153.3, 158.0, 158.2, 158.9, 162.2, 165.5, 167.5; HRMS (ESI) calcd for [MH] - C 30 H 27 N4O6S - : 571.1657 found 571.1661. Example 4: Preparation of (E)-4-(2-hydroxy-3-((4-((4-methylbenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)propoxy)-2H-benzopyran-2-one (compound 10d) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 4-methylbenzaldehyde.

[0046] The structural formula of compound 10d is shown below:

[0047] Compound 10d was a white powder with a yield of 45%; melting point: 138.5-141.9℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10d are shown below: 1 H NMR (DMSO, 400 MHz) d : 2.32 (s, 3H), 3.48 (d, J = 4.4 Hz, 1H), 4.15-4.23 (m, 3H), 5.24 (s, 2H), 5.76 (d, J= 4.0 Hz, 1H), 5.82 (s, 1H), 6.90-6.99(m, 3H), 7.22-7.37 (m, 6H), 7.60- 7.68 (m, 3H), 7.88 (d, J = 8.0 Hz, 1H), 8.82 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 21.8, 35.9, 60.1, 67.6, 72.5, 91.1, 115.5, 115.7, 116.9, 122.1, 123.8, 124.6, 129.5, 129.5, 130.1, 130.3, 133.3, 144.3, 148.6, 149.5, 153.3, 157.9, 162.2, 165.4, 166.7; HRMS (ESI) calcd for [M+H] + C 29 H 27 N4O5S + : 543.1697 found 543.1690. Example 5: Preparation of ((E)-4-(3-((4-((3-chloro-4-hydroxybenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)-2-hydroxypropoxy)-2H-benzopyran-2-one (compound 10e) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 3-chloro-4-hydroxybenzaldehyde.

[0048] The structural formula of compound 10e is shown below:

[0049] Compound 10e is a white powder with a yield of 48%; melting point: 152.3-153.9 ℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10e are shown below: 1 H NMR (DMSO, 400 MHz) d 3.47 (d, J= 4.8 Hz, 2H), 4.15-4.23 (m, 3H), 5.22 (s, 2H), 5.76 (m, 2H), 6.90-7.05 (m, 4H), 7.22-7.37 (m, 4H), 7.60-7.63(m, 2H), 7.75 (s, 1H), 7.87 (d, J = 8.0 Hz, 1H), 8.71 (s, 1H), 11.29 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 35.9, 60.1, 67.6, 72.5, 91.2, 115.5, 115.7, 116.9, 117.6, 121.1, 122.1, 123.8, 124.1, 124.6, 129.9, 130.1, 131.3, 133.3, 149.3, 153.3, 157.9, 158.2, 162.2, 165.5, 165.6; HRMS (ESI) calcd for [M+H] + C 28 H 24 ClN4O6S + : 579.1100 found 579.1097. Example 6: Preparation of (E)-4-(3-((4-((4-fluorobenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)-2-hydroxypropoxy)-2H-benzopyran-2-one (compound 10f) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 4-fluorobenzaldehyde.

[0050] The structural formula of compound 10f is shown below:

[0051] Compound 10f is a white powder with a yield of 48%; melting point: 133.6-135.1℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10f are shown below: 1 H NMR (DMSO, 400 MHz) d 3.52 (d, J= 4.4 Hz, 2H), 4.15-4.23 (m, 3H), 5.26 (s, 2H), 5.72-5.84 (m, 2H), 6.90-6.99 (m, 3H), 7.22-7.37 (m, 6H), 7.60-7.64 (m, 1H), 7.85-7.89 (m, 3H), 8.88 (s, 1H); 13 C NMR (100 Hz, DMSO) d :35.9, 60.1, 67.5, 72.5, 91.2, 115.5, 115.7,116.9, 117.0, 117.2, 122.2, 123.8, 124.7, 128.8, 130.1, 132.0 (d, J = 10.0 Hz,2C), 133.4, 148.6, 149.6, 153.3, 157.9, 162.2, 164.2, 165.3 (d, J = 11.0 Hz, 2C), 166.7; HRMS (ESI) calcd for [M+H] + C 28 H 24 FN4O5S + : 547.1446 found 547.1439. Example 7: Preparation of (E)-4-(3-((4-((2-fluoro-4-hydroxybenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)-2-hydroxypropoxy)-2H-benzopyran-2-one (compound 10 g) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 2-fluoro-4-hydroxybenzaldehyde.

[0052] The structural formula of 10g of compound is shown below:

[0053] The compound 10g was a light yellow powder with a yield of 57%; melting point: 183.9-185.4℃; and the data of the 1H NMR, 1C NMR, and high-resolution mass spectrometry of the compound 10g are shown below: 1 H NMR (DMSO, 400 MHz) d 3.49 (d, J= 13.2 Hz, 2H), 4.17-4.23 (m, 3H), 5.20 (s, 2H), 5.76 (s, 1H), 5.83 (d, J = 3.2 Hz, 2H), 6.61 (d, J = 12.4 Hz, 1H), 6.70 (d, J = 8.8 Hz, 1H), 6.90-6.99 (m, 3H), 7.22-7.36 (m, 4H), 7.59-7.63 (m,1H), 7.76-7.88 (m, 2H), 8.84 (s, 1H), 10.97 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 36.0, 60.2, 67.7, 72.5, 91.2, 103.6, 110.9, 113.8, 115.6, 115.8, 116.9, 122.2, 123.8, 124.6, 129.9, 130.1, 133.3, 148.6, 149.4, 153.3, 157.9, 159.8, 162.1, 162.3, 164.6, 164.7, 165.2, 165.5; HRMS (ESI) calcd for [M+H] + C 28 H 24 FN4O6S + : 563.1395 found 563.1396. Example 8: Preparation of (E)-4-(2-hydroxy-3-((4-((4-hydroxy-2,6-dimethylbenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)propoxy)-2H-benzopyran-2-one (compound 10h) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 4-hydroxy-2,6-dimethylbenzaldehyde.

[0054] The structural formula of compound 10h is shown below:

[0055] Compound 10h was a white powder with a yield of 53%; melting point: 184.5-186.7℃; and the data of compound 10h's 1H NMR, 1C NMR, and high-resolution mass spectrometry are shown below: 1 H NMR (DMSO, 400 MHz) d :2.34 (s, 6H), 3.49 (d, J = 13.2 Hz, 2H), 4.18-4.24 (m, 3H), 5.19 (s, 2H), 5.75 (s, 1H), 5.83 (s, 1H), 6.51 (s, 2H), 6.91-6.99 (m, 3H), 7.22-7.37 (m, 4H), 7.60-7.64 (m, 1H), 7.87 (d, J = 8.4 Hz, 1H), 8.97 (s, 1H), 10.13 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 22.3, 35.7, 59.9, 67.6, 72.5, 91.1, 115.2, 115.7, 116.7, 116.9, 120.1, 122.0, 123.8, 124.6, 130.1, 133.3, 143.2, 148.4, 149.2, 153.3, 158.0, 160.9, 162.2, 165.5, 165.9; HRMS (ESI) calcd for [M+H] + C 30 H 29 N4O6S + : 573.1802 found 573.1800. Example 9: Preparation of (E)-4-(2-hydroxy-3-((4-((4-hydroxy-3-nitrobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)propoxy)-2H-benzopyran-2-one (compound 10i) Compared to Example 1, the difference is that only 3-nitro-4-hydroxybenzaldehyde is replaced with 3-hydroxybenzaldehyde.

[0056] The structural formula of compound 10i is shown below:

[0057] Compound 10i is a yellow powder with a yield of 51%; melting point: 100.3-102.1 ℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10i are shown below: 1 H NMR (DMSO, 400 MHz) d 3.48 (d, J = 4.4 Hz, 2H), 4.15-4.23 (m, 3H), 5.26 (s, 2H), 5.75 (d, J = 5.2 Hz, 1H), 5.83 (s, 1H), 6.89-6.99 (m, 3H), 7.16-7.25 (m, 3H), 7.29-7.36 (m, 2H), 7.59-7.63 (m, 1H), 7.87 (d, J = 8.4 Hz, 1H), 7.94 (d, J = 8.8 Hz, 1H), 8.26 (s, 1H), 8.82 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 35.5, 59.8, 67.2, 72.1, 90.8, 115.2, 115.3, 116.5, 120.5, 121.7, 122.6, 123.4, 124.2, 127.5, 129.7, 132.9, 134.0, 137.3, 148.3, 149.0, 152.9, 156.4, 157.5, 161.7, 164.2, 165.1; HRMS (ESI) calcd for [M+H] + C 28 H 24 N5O8S + : 590.1340 found 590.1340. Example 10: Preparation of (E)-4-(3-((4-((4-chlorobenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)-2-hydroxypropoxy)-2H-benzopyran-2-one (compound 10j) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 4-chlorobenzaldehyde.

[0058] The structural formula of compound 10j is shown below:

[0059] Compound 10j was a white powder with a yield of 62%; melting point: 168.4-170.2 ℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10j are shown below: 1 H NMR (DMSO, 400 MHz) d : 3.94-3.98 (m, 1H), 4.24-4.41 (m, 1H), 4.85-4.67 (m, 3H), 5.71 (s, 2H), 6.21-6.24 (m, 2H), 7.31-7.42 (m, 3H), 7.63-7.75(m, 4H), 7.94-8.03 (m, 3H), 8.19-8.29 (m, 3H), 9.30 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 35.5, 46.2, 46.4, 51.2, 59.8, 67.1, 72.0, 90.7, 115.2, 116.4, 121.7, 123.3, 124.1, 129.4, 129.6, 130.5, 137.9, 148.1, 149.3, 152.8, 157.4, 161.7, 164.2, 164.9; HRMS (ESI) calcd for [M+H] + C 28 H 24 ClN4O5S + : 563.1150 found 563.1174. Example 11: Preparation of (E)-4-(3-((4-((4-bromobenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)-2-hydroxypropoxy)-2H-benzopyran-2-one (compound 10k) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 4-bromobenzaldehyde.

[0060] The structural formula of compound 10k is shown below:

[0061] Compound 10k is a yellowish-brown powder with a yield of 62%; melting point: 139.2-142.7 ℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10k are shown below: 1 H NMR (DMSO, 400 MHz) d : 3.97-3.99 (m, 2H), 4.64-4.69 (m, 3H), 5.75(s, 2H), 6.24-6.28 (m, 2H), 7.37-7.45 (m, 3H), 7.68-7.80 (m, 4H), 8.05-8.33(m, 6H), 9.32 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 35.5, 46.2, 46.4, 51.2, 59.8, 67.1, 72.1, 90.7, 115.1, 115.2, 116.4, 121.7, 123.3, 124.1, 127.0, 129.7, 130.7, 132.4, 132.8, 148.1, 149.3, 152.8, 157.4, 161.7, 164.4, 165.0; HRMS (ESI) calcd for [M+H] + C 28 H 24 BrN4O5S + : 607.0645 found 607.0660. Example 12: Preparation of (E)-4-(2-hydroxy-3-((4-((4-hydroxy-3-methoxy-5-nitrobenzylidene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)propoxy)-2H-benzopyran-2-one (compound 10l) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 3-bromo-4-hydroxy-5-nitrobenzaldehyde.

[0062] The structural formula of compound 10l is shown below:

[0063] Compound 10l is a yellow oily substance with a yield of 47%; melting point: 158.6-159.2 ℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10l are shown below: 1 H NMR (DMSO, 400 MHz) d: 3.49-3.54 (m, 2H), 5.27 (s, 2H), 5.76-5.83(m, 2H), 6.91 (t, J = 3.2 Hz, 1H), 6.98 (d, J = 7.2 Hz, 2H), 7.21-7.36 (m, 5H), 7.54-7.63 (m, 2H), 7.86-7.89 (m, 2H), 8.80 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 36.1, 57.2, 60.2, 67.6, 72.5, 91.2, 112.7, 115.6, 116.9, 120.4, 121.9, 122.1, 123.8, 124.6, 130.1, 133.3, 137.5, 148.0, 148.8, 149.2, 150.8, 153.3, 158.0, 162.1, 164.9, 165.5; HRMS (ESI) calcd for [MH] - C 29 H 24 N5O9S - : 618.1300 found 618.1298. Example 13: Preparation of (E)-4-(3-((4-((3-chlorobenzyl)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)-2-hydroxypropoxy)-2H-benzopyran-2-one (compound 10m) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 3-chlorobenzaldehyde.

[0064] The structural formula of compound 10m is shown below:

[0065] Compound 10m is a white powder with a yield of 62%; melting point: 168.5-171.1 ℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10m are shown below: 1 H NMR (DMSO, 400 MHz) d :3.34-3.54 (m, 2H), 4.16-4.24 (m, 3H), 5.31(s, 2H), 5.76 (d,J = 5.2 Hz, 1H), 5.84 (s, 1H), 6.91-7.01 (m, 3H), 7.22-7.37(m, 4H), 7.50-7.54 (m, 1H), 7.62-7.66 (m, 2H), 7.74-7.78 (m, 2H), 7.87 (d, J =9.2 Hz, 1H), 8.88 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 36.2, 60.4, 67.7, 72.5, 91.2, 115.6, 115.7, 115.8, 116.5, 116.9, 122.2, 122.6, 123.8, 124.6, 128.0, 128.7, 130.1, 131.7, 133.3, 134.3, 134.5, 148.8, 149.7, 153.3, 158.0, 162.1, 164.4, 165.5; HRMS (ESI) calcd for [M+H] + C 28 H 23 ClN4O5S + : 563.1150 found 563.1151. Example 14: Preparation of (E)-4-(3-((4-(([1,1'-biphenyl]-4-ylmethylene)amino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-yl)thio)-2-hydroxypropoxy)-2H-benzopyran-2-one (compound 10n) Compared to Example 1, the only difference is that 3-hydroxybenzaldehyde is replaced with 4-biphenylbenzaldehyde.

[0066] The structural formula of compound 10n is shown below:

[0067] Compound 10n is a yellow powder with a yield of 58%; melting point: 125.4-127.8 ℃; and the 1H NMR, 1C NMR, and high-resolution mass spectrometry data of compound 10n are shown below: 1 H NMR (DMSO, 400 MHz) d:3.50-3.55 (m, 2H), 4.17-4.24 (m, 3H), 5.28(s, 2H), 5.83-5.86 (m, 2H), 6.90-7.01 (m, 2H), 7.22-7.40 (m, 6H), 7.44-7.48(m, 2H), 7.60 (t, J = 7.6 Hz, 1H), 7.70 (d, J = 7.6 Hz, 2H), 7.77-7.79 (m, 2H), 7.86-7.90 (m, 3H), 8.93 (s, 1H); 13 C NMR (100 Hz, DMSO) d : 36.2, 60.4, 67.7, 72.5, 91.2, 115.7, 115.8, 116.5, 116.9, 122.2, 122.6, 123.8, 124.6, 128.0, 128.7, 130.1, 131.7, 133.3, 134.3, 134.5, 148.8, 149.7, 153.3, 158.0, 162.1, 164.4, 165.5; HRMS (ESI) calcd for [M+H] + C 34 H 28 N4O5S + : 605.1853 found 605.1852. To further verify the superior effects of the present invention, the inventors also conducted the following comparative experiments: Experiment 1 Different concentrations of the compound or acarbose (10 μL) and 150 μL of α-glucosidase solution (0.1 U / mL) were added to 96-well plates, and the mixture was incubated at 37°C for 15 minutes. Then, 40 μL of p-nitrophenyl-α-D-glucopyranoside (1.25 mM) was added to the above mixture, and after incubation for another 30 minutes, the samples were detected using a microplate reader at a wavelength of 405 nm. The IC50 was calculated. 50 The results are shown in Table 1.

[0068] Table 1. Inhibitory activity of α-glucosidase by 1,2,4-triazole-coumarins (IC50) 50 )

[0069] As can be seen from Table 1, most of the compounds synthesized in this invention exhibit good α-glucosidase inhibitory activity, with compound 10n exhibiting the strongest α-glucosidase activity, its IC50 value being [missing value]. 50 The concentration was 9.71 ± 0.28 μM, which was superior to the positive control drug acarbose (309.83 ± 8.74 μM).

[0070] Experiment 2 The compound 10n, exhibiting the strongest α-glucosidase activity, was dissolved in 10% DMSO. The final concentration of DMSO in all reaction systems was controlled to be 1%. PTP1B and compound 17 were reacted at room temperature for 30 min in a 50 μL reaction system containing 25 mM MOPS (pH=7.0), 50 mM NaCl, 0.05% Tween 20, 3 mM DTT, and 10 μM DiFMUP. Fluorescence intensity was measured using a Tecan Infinite M1000 microplate reader at an excitation wavelength of 358 nm and an emission wavelength of 455 nm. Phosphatase activity assays were performed repeatedly at each concentration, and fluorescence intensity data were analyzed using a GrapHpad Prism8. The results indicate that compound 10n possesses certain PTP1B inhibitory activity, with an IC50 value of [missing value]. 50 The value was 5.76 ± 0.15 μM, and the control IC50 for ursolic acid was [value missing]. 50 The value was 5.62 ± 0.22 μM.

[0071] Experiment 3 Compound 10n was used to investigate its effect on postprandial blood glucose in normal Kunming mice.

[0072] In this experiment, Kunming mice were first acclimatized for one week and then randomly divided into four groups (blank control group, negative control group, drug group, and positive control acarbose group), with eight mice in each group. The mice were fasted but allowed free access to water for 12 hours. Before the experiment, the test sample 14 (4 mg / mL), acarbose (2 mg / mL), and sucrose (0.7 g / mL) were uniformly suspended in 0.5% CMC-Na aqueous solution under ultrasound. Each mouse in each group was numbered and weighed. The compound group and positive control group were then administered the compound via gavage at a volume of 20 mg / kg. Forty minutes later, based on the mice's body weight (2.5 g / kg), the blank control group was administered a pre-prepared sucrose solution via gavage, and the negative control group was administered a 0.5% CMC-Na aqueous solution. Blood glucose concentrations in the tail vein of the mice were monitored and recorded at 0, 15, 30, 60, 90, and 150 minutes after glucose administration using a Roche Accu-Chek Instant glucometer. The results were processed using Origin 64 software. All procedures involving animals were strictly followed and adhered to the "Guidelines for Laboratory Animal Welfare and Ethics." Furthermore, these experiments have been approved by the Laboratory Animal Management and Ethics Committee of Guizhou Medical University (Ethical Guidelines: 2001117).

[0073] Depend on Figure 2 (A) It can be seen that after oral administration of sucrose, the blood glucose level in the model group rose rapidly and reached a peak, while the blood glucose level in the oral acarbose group and the compound 10n group was significantly lower than that in the model group after oral administration of sucrose, and the blood glucose level decreased steadily. Figure 2 (B) describes the changes in glycemic load in mice after oral administration of sucrose. The figure shows that compound 10n can significantly reduce postprandial blood glucose levels.

[0074] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B, characterized in that, It has a general structural formula as shown in equation (I): ； R is selected from one of 3-hydroxy, 4-hydroxy, 4-hydroxy-3,5-dimethyl, 3-chloro-4-hydroxy, 4-fluoro, 2-fluoro-4-hydroxy, 4-hydroxy-2,6-dimethyl, 3-nitro-4-hydroxy, 3-bromo-4-hydroxy-5-nitro, 3-chloro, and 4-biphenyl.

2. A method for preparing a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B as described in claim 1, characterized in that, The method specifically includes the following steps: Step 1: Add 4-hydroxycoumarin, epichlorohydrin, and sodium hydroxide, followed by anhydrous ethanol as solvent, and reflux at 60-100℃ for 4-10 hours; after the reaction is complete, purify by column chromatography to obtain the product 4-(epoxyethylene-2-ylmethoxy)-2H-benzopyran-2-one. Step 2: Phenol and chloroacetic acid are reacted in an ethanol solution of sodium hydroxide at 60-100℃ for 4-10h. After the reaction is completed, hydrochloric acid is added to adjust the pH to 2-3. The mixture is allowed to stand at 4℃ to crystallize and then filtered to obtain the product 2-phenoxyacetic acid. Step 3: Reflux the product 2-phenoxyacetic acid with aminothiourea at 120-160℃ for 1-3 h, wash the reaction mixture with H2O, filter and dry to obtain the filter residue, which is the product 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol. Step 4: The product 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol is reacted with benzaldehyde with different substituents in glacial acetic acid solvent at 100-140℃ for 2-8 hours. After standing, the precipitate is collected by filtration to obtain the residue, which is (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol with different substituents. Step 5: The obtained (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol, 4-(ethylene oxide-2-ylmethoxy)-2H-benzopyran-2-one, and anhydrous ethanol with different substituents are refluxed at 60-100℃ for 4-10 h; after the reaction is completed, the target compound I is obtained by column chromatography purification, namely the 1,2,4-triazol-coumarin α-glucosidase and PTP1B dual-target inhibitor.

3. The preparation method according to claim 2, characterized in that, In step 1, the ratio of 4-hydroxycoumarin, epichlorohydrin, sodium hydroxide, and anhydrous ethanol is 18-56 mmol: 235-707 mmol: 20-62 mmol: 150-450 mL, and the eluent for silica gel column chromatography is petroleum ether: ethyl acetate = 3:

1.

4. The preparation method according to claim 2, characterized in that, In step 2, the ratio of phenol, chloroacetic acid, sodium hydroxide and anhydrous ethanol is 21-84 mmol: 38-152 mmol: 127-508 mmol: 15-60 mL.

5. The preparation method according to claim 2, characterized in that, In step 3, the molar ratio of 2-phenoxyacetic acid to aminothiourea is 1-100 mmol: 1-100 mmol.

6. The preparation method according to claim 2, characterized in that, In step 4, the ratio of benzaldehyde with different substituents, 4-amino-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol and glacial acetic acid is 1-100 mmol: 1.2-120 mmol: 6-600 mL.

7. The preparation method according to claim 2, characterized in that, In step 5, the ratio of (E)-4-(benzylideneamino)-5-(phenoxymethyl)-4H-1,2,4-triazol-3-thiol, 4-(ethylene oxide-2-ylmethoxy)-2H-benzopyran-2-one and anhydrous ethanol with different substituents is 1-100 mmol: 1-100 mmol: 35-3500 mL.

8. The use of a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B as described in claim 1, or a dual-target inhibitor of 1,2,4-triazole-coumarin α-glucosidase and PTP1B prepared by any one of claims 2 to 7, in the preparation of antidiabetic drugs.

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