Application of Glucosinolates in the Preparation of Hypoglycemic Drugs and Health-Related Products

Abstract

This invention discloses the application of aromatic thioglycosides in the preparation of hypoglycemic drugs and health-related products. Aromatic thioglycosides exhibit good inhibitory activity against α-glucosidase. The thioglycoside compounds provided by this invention show promise for the prevention or treatment of diabetes.

Description

Technical Field

[0001] This invention belongs to the technical field of hypoglycemic drugs or health-related products, specifically relating to the application of sulforaphane in the preparation of hypoglycemic drugs and health-related products. Background Technology

[0002] Diabetes is a metabolic disorder caused by a variety of factors, mainly manifested as a syndrome of lipid and glucose metabolism disorders. To date, there is no effective drug or method to cure it; the only approach is to control blood sugar within a certain range to delay the onset of complications. Therefore, the development of new therapeutic drugs is urgently needed.

[0003] Glucosides are important secondary metabolites in plants. Due to their significant impact on both plants and humans, this field has become a research hotspot in biology. Glucoside compounds have wide applications in various fields, primarily including energy storage and signal transduction in organisms, anticancer effects, and antibacterial effects. Glucoside compounds also have important applications in drug development. We synthesized a series of glutoside compounds using a simple and efficient synthetic method, and then tested the in vitro alpha-glycosidase activity of all synthesized compounds, hoping to provide new insights for the development of hypoglycemic drugs and health-related products.

[0004] The applicant conducted research on the synthesis of thioglycosides. During the research, the applicant experimentally determined that thioglycosides 1-5 had an inhibitory effect on alpha-glycosidase. Summary of the Invention

[0005] The purpose of this invention is to provide the application of sulfoglycosides in the preparation of hypoglycemic drugs and health-related products, characterized in that the drugs contain sulfoglycoside compounds.

[0006] α-Glucosidase is α-glucosidase.

[0007] The structure of the aromatic thioglycoside compound is as follows:

[0008]

[0009] Advantages of this invention:

[0010] This invention provides the application of aromatic thioglycosides in the preparation of hypoglycemic drugs and health-related products. Thioglycoside compounds exhibit good inhibitory activity against α-glucosidase. The method for preparing thioglycoside compounds provided by this invention is simple and holds promise for further addressing the problem of diabetes. Attached Figure Description

[0011] Figure 1 The structural formula is that of an aromatic thioglycoside compound;

[0012] Figure 2The synthetic routes for compounds 1 and 2 are shown below.

[0013] Figure 3 The synthetic routes for compounds 3-5 are shown below. Detailed Implementation

[0014] The present invention will be further described below, but this is not intended to limit the invention in any way. Any modifications made based on the present invention are within the scope of protection of the present invention.

[0015] This provides information on the application of glucosinolates in the preparation of hypoglycemic drugs and health-related products, which contain glucosinolate compounds.

[0016] α-Glucosidase is α-glucosidase.

[0017] The structural formula of the thioglycoside compound is shown in [reference needed]. Figure 1 .

[0018] Preparation of thioglycosides:

[0019] (1) Synthesis of p-Tolyl-2,3,4,6-Tetra- O -Acetyl-1-thio- β -D-Gluconopyranoside 1: Starting from D-glucose, after full acetylation, compound 1 was synthesized by reacting it with p-toluene in the presence of Lewis acid boron trifluoride diethyl ether. The synthetic route is shown below. Figure 2 .

[0020] (2) Synthesis of p-Tolyl-2,3,4,6-Tetra- O -benzyl-1-thio- β -D-Glucopyranoside 2: Starting from compound 1, its hydrolysis was followed by BnBr protection of the hydroxyl groups to yield compound 2. The synthetic route is shown below. Figure 2 .

[0021] (3) Synthesis of p-Tolyl-2- O -Acetyl-3,4,6-tri- O -Benzyl-1-thioglycoside derivatives 3-5: Sugars with different configurations were fully acetylated and brominated at the 1-position. The brominated sugars were reacted with dry acetonitrile, anhydrous ethanol, TBAI, and TEA under reflux at 60°C to obtain orthoesterified ester products. The acetyl groups at positions 3, 4, and 6 were then reacted with BnBr, KOH, and reflux at 70°C to generate orthoesterified ester products protected by benzyl groups at positions 3, 4, and 6. p-Toluenethiophenol was added, dissolved in DCM, and the mixture was reacted dropwise with boron trifluoride diethyl ether under N2 protection to obtain compounds 3-5. For detailed synthetic routes, see [link to synthetic route]. Figure 3 .

[0022] Example 1

[0023] Synthesis method of compound 2

[0024]

[0025] In a reaction flask, NaOAc (1 g, 12.2 mmol) and Ac₂O (9.4 mL, 99.4 mmol) were added, and the mixture was heated to reflux. D-glucose (1.8 g, 10 mmol) was added in portions, and the mixture was stirred under reflux (TLC monitoring). The reaction solution was cooled to room temperature, crushed ice was added, and the mixture was sonicated to precipitate a white solid. The solid was filtered, washed with water until no acidic odor remained, and the filter cake was dissolved with hot EtOH. PE was then added, and the mixture was allowed to stand and cool to precipitate a solid. The solid was filtered, and the filter cake was washed with PE to obtain 3.8 g of white solid, with a yield of 97%. In a dry round-bottom flask, the white solid (2 g, 5.1 mmol), 2 g of 4Å molecular sieve, and p-toluenethiophenol (1.3 g, 10.5 mmol) were added and dissolved in dry DCM (20 mL). The mixture was cooled to 0 °C, and BF₃·OEt₂ (2.2 mL, 17.5 mmol) was slowly added dropwise under N₂ protection. After the addition was complete, the mixture was brought to room temperature and stirred overnight (TLC monitoring). Wash with saturated Na₂CO₃ solution, extract with DCM, combine the organic layers, wash with saturated NaCl solution, dry with anhydrous Na₂SO₄, filter, concentrate the filtrate, and perform silica gel column chromatography (eluent: PE / EA = 4 / 1). V / V Purification yielded a white solid 1 (p-Tolyl-2,3,4,6-tetra- O -Acetyl-1-thio- β 1.8 g of β-D-glucopyranoside (78% yield). mp 113.9~117.6℃.

[0026] Dissolve 1 (2 g, 4.4 mmol) in MeOH (25 mL), add NaOMe (113 mg, 2.1 mmol), stir at room temperature, and monitor the reaction until complete by TLC. Remove the solvent under reduced pressure. Add DMF (25 mL), cool to 0 °C, add NaH (1.8 g, 44 mmol, 60%) in small amounts several times, stir for 0.5 h, add BnBr (5.2 mL, 44 mmol), react overnight at room temperature, and monitor the reaction until complete by TLC. Quench the reaction with water (5 mL), dilute the reaction solution with EA, wash with water and saturated NaCl solution, dry with anhydrous Na2SO4, filter, concentrate the filtrate, and precipitate the residue by silica gel column chromatography (eluent: PE / EA = 35 / 1 ~ PE / EA = 14 / 1). V / V ), yielding a white solid 2 (p-tolyl-2,3,4,6-tetra- O -benzyl-1-thio- β1.99 g of β-D-glucopyranoside (70.1% yield) was obtained. mp 95.6–97.7 °C.

[0027] The synthesis methods of compounds 3-5 are explained, taking the synthesis of compound 3 as an example.

[0028]

[0029] Add D-galactose (4 g, 22.2 mmol), Ac₂O (20 mL, 211.6 mmol), and I₂ (180 mg, 0.71 mmol) to a reaction flask, and stir at room temperature until the reaction system becomes a brown transparent state. After the reaction is complete, add dry DCM (100 mL), cool the reaction solution to 0 °C, add 33% HBr-AcOH solution (42 mL) dropwise, stir the reaction, remove the ice-water bath, and stir the reaction at room temperature for 1 h (TLC monitoring of the reaction). After the reaction is complete, dilute the reaction solution with DCM, wash with ice water and sodium thiosulfate aqueous solution, dry the organic layer with anhydrous Na₂SO₄, filter, concentrate the filtrate, and obtain 7.5 g of colorless oily crude product. Take the crude product (5 g, 12.2 mmol) and add TBAI (395 mg, 1.22 mmol), dry acetonitrile (50 mL), MeOH (0.98 mL, 24.3 mmol), and TEA (3.4 mL, 24.3 mmol). React at 60 °C for 1 h (TLC monitoring of the reaction). Dilute the reaction solution with EA, wash successively with water and saturated NaCl solution, dry with anhydrous Na₂SO₄, filter, concentrate the filtrate under reduced pressure, and perform silica gel column chromatography (eluent: PE / EA = 4 / 1). V / V ), yielding 4.1 g of a colorless oily liquid. A portion of this (3.6 g, 9.9 mmol) was dissolved in MeOH (36 mL), and MeONa (113 mg, 2.1 mmol) was added. The mixture was stirred at room temperature for 1 h. After the reaction was complete, the concentrated reaction solution was evacuated under a diaphragm vacuum pump for 0.5 h. DMF (80 mL) was added to dissolve the solution, and NaH (3.96 g, 99 mmol, 60%) was added at 0 °C. The mixture was stirred until no bubbles were generated, and then BnBr (11.8 mL, 99 mmol) was added. The mixture was stirred at room temperature overnight (TLC detection of the reaction). After the reaction was complete, MeOH (50 mL) was added to quench the reaction. The mixture was distilled under reduced pressure, and the residue was diluted with Et2O (300 mL). The residue was washed with water, and the aqueous layer was extracted with EA (75 mL × 2). The organic layers were combined, washed with saturated NaCl solution (300 mL), and dried over anhydrous Na2SO4. Silica gel column chromatography (eluent: PE / EA = 10 / 1) was performed. V / V), yielding a yellow oily liquid (methyl 1,2-orthoacetate-3,4,6-tri- O -Benzyl- α 3.4 g of β-D-galactopyranoside (D-galactopyranoside), yield 51%.

[0030] A yellow oily liquid (1.9 g, 3.6 mmol) was dissolved in 60% AcOH (30 mL). The mixture was stirred at room temperature for 4 h. After the reaction was complete, the reaction solution was evaporated to dryness and then rinsed three times with toluene (10 mL). Pyr (30 mL) was added to the crude product to dissolve it, followed by the addition of DMAP (88 mg, 0.7 mmol). The mixture was cooled to 0 °C, and Ac2O (0.7 mL, 7.2 mmol) was slowly added dropwise under N2 protection. The mixture was then brought to room temperature and stirred overnight. After the reaction was complete, the reaction system was cooled to 0 °C, and anhydrous ethanol (1 mL) was added to quench the reaction. The reaction solution was evaporated to dryness, and then dissolved in EA (60 mL). It was washed with 1 M HCl (250 mL), saturated sodium bicarbonate solution (250 mL), saturated NaCl solution (250 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated to dryness. The crude product was purified to give 1.5 g of yellow oily liquid, with a yield of 81%.

[0031] A yellow oily substance (1.5 g, 2.9 mmol) and p-toluenethiophenol (0.7 g, 5.8 mmol) were dissolved in dry DCM (20 mL). The solution was placed in an ice-water bath, and 48% BF3·OEt2 (0.4 mL, 4.3 mmol) was slowly added dropwise under N2 protection. The mixture was then brought to room temperature and stirred for 4 h. The reaction was monitored by TLC. The reaction solution was diluted with dichloromethane, and the organic layer was washed with saturated sodium bicarbonate solution, water, and saturated NaCl solution, and dried over anhydrous Na2SO4. The solution was filtered, and the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography to obtain compound 3 (p-tolyl-2- O -Acetyl-3,4,6-tri- O -benzyl-1-thio- β -D-galactopyranoside), white solid, 0.7 g, yield 52%.

[0032] p-Tolyl-2- O -Acetyl-3,4,6-tri- O -benzyl-1-thio- β -D-glucopyranoside (4): white solid, 0.6 g, yield 35%. 1 H NMR (600 MHz, CDCl3) δ 7.48 – 7.00 (m, 19H), 4.98 (t,J =9.4 Hz, 1H), 4.78 (dd, J = 11.2, 8.4 Hz, 2H), 4.66 (d, J = 11.4 Hz, 1H), 4.61 –4.50 (m, 4H), 3.78 (dd, J = 11.0, 2.1 Hz, 1H), 3.73 (dd, J = 11.1, 4.9 Hz, 1H), 3.70 – 3.63 (m, 2H), 2.30 (s, 3H), 2.00 (s, 3H). 13 C NMR (150 MHz, CDCl3) δ 169.6, 138.4, 138.2, 138.2, 138.0, 133.3, 129.7, 128.9, 128.6, 128.5, 128.2, 128.0, 127.9, 127.8, 127.7, 86.2, 84.6, 79.6, 77.9, 75.4, 75.2, 73.6, 72.0, 69.1, 21.3, 21.2.

[0033] p-Tolyl-2- O -Acetyl-3,4,6-tri- O -benzyl-1-thio- β -D-Mannopyranoside (5): colorless oily liquid, 0.7 g, yield 52%. 1 H NMR (600 MHz, CDCl3) δ 7.40 – 7.00 (m, 19H), 5.59 (s,1H), 5.45 (s, 1H), 4.88 (d, J = 10.9 Hz, 1H), 4.72 (d, J = 11.3 Hz, 1H), 4.65 (d, J = 12.0 Hz, 1H), 4.56 (d, J = 11.4 Hz, 1H), 4.51 (d, J = 10.7 Hz, 1H), 4.46 (d, J =12.0 Hz, 1H), 4.34 (q, J = 5.2 Hz, 1H), 3.94 (d, J = 6.2 Hz, 2H), 3.85 (dd,J =10.9, 4.7 Hz, 1H), 3.72 (d, J = 11.3 Hz, 1H), 2.30 (s, 3H), 2.13 (s, 3H).

[0034] Example 2

[0035] Alpha-glycosidase activity assay

[0036] I. Materials and Methods

[0037] 1. Drugs and samples

[0038] Thioglycoside compounds 1-5 were prepared according to Example 1.

[0039] All test samples were dissolved in DMSO and stored in a 4°C refrigerator.

[0040] 2. Experimental Methods

[0041] Add 112 μL of potassium phosphate buffer (pH 6.8) and 20 μL of 0.2 U / mL solution to a 96-well plate. -1 α-Glucosidase (prepared with potassium phosphate buffer), 8 μL DMSO, 8 μL of different concentrations (220.00, 110.00, 55.00, 27.50, 13.75 μmol·L⁻¹). -1 The sample solution was incubated at 37°C for 15 min, and then 20 μL of 2.5 mmol·L⁻¹ solution was added. -1 Add PNPG (prepared with potassium phosphate buffer), shake well, and react at 37°C for 15 min. Then add 80 μL of 0.2 mol·L⁻¹. -1 The reaction was terminated with Na2CO3 solution, and the absorbance was measured at a wavelength of 405 nm. Each sample was measured in triplicate.

[0042] Sample determination: There were 4 groups in total, with 3 replicates per group and a total volume of 240 μL per well. The groups were: a: blank group (8 μL DMSO + 112 μL potassium phosphate buffer + 20 μL enzyme solution + 20 μL PNPG + 80 μL Na2CO3); b: blank control group (8 μL DMSO + 152 μL potassium phosphate buffer + 80 μL Na2CO3); c: sample determination group (8 μL sample + 112 μL potassium phosphate buffer + 20 μL enzyme solution + 20 μL PNPG + 80 μL Na2CO3); d: sample control group (8 μL sample + 152 μL potassium phosphate buffer + 80 μL Na2CO3).

[0043] 3. Calculation formula

[0044] According to I%=[1- (A c -A d ) / (A a -A b The suppression rate is calculated by multiplying the value by 100%, and the corresponding IC is obtained using Origin 8.0 software. 50 value.

[0045] II. Results

[0046]

[0047] III. Conclusion

[0048] 1. Samples that showed activity against α-glucosidase as a single drug include: 3 and 4.

[0049] In summary, glucosinolate compounds 3-4, when used alone, have an inhibitory effect on α-glucosidase, and the inhibitory effect is good.

Claims

1. Use of an aromatic thioglycoside for the preparation of an α-glucosidase inhibitor, characterized in that, The structure of the aromatic glucosinolate is: 。 2. A hypoglycemic agent, characterized by An aromatic glucosinolate compound comprising the structure of claim 1.

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