Application of aromatic thioglycoside in preparation of hypoglycemic drugs and health-care related products

Abstract

The invention discloses application of aromatic thioglycoside in preparation of hypoglycemic drugs and health-care related products. The aromatic thioglycoside compound has a good inhibitory activity effect on alpha-glycosidase. The thioglycoside compound provided by the invention is expected to be used for preventing or treating diabetes mellitus.

Description

Technical Field

[0001] The invention belongs to the technical field of hypoglycemic drugs or health-related products, and specifically relates to the application of aromatic glucosinolates in the preparation of hypoglycemic drugs and health-related products. Background Art

[0002] Diabetes is a metabolic disorder caused by multiple reasons, mainly manifested as lipid metabolism and sugar metabolism disorder syndrome. So far, there is no effective drug or method to cure it. It can only control blood sugar within a certain range to delay the occurrence of complications. Therefore, the development of new therapeutic drugs is urgent.

[0003] Glucoside is an important secondary metabolite in plants. Since it has a great impact on both plants and humans, this field has become a research hotspot in the biological world. Glucoside compounds have a wide range of applications in many fields, mainly including energy storage and signal transduction in organisms, anti-cancer effects, antibacterial effects, etc. Glucoside compounds also have important applications in the field of drug research and development. We use a simple and efficient synthesis method to synthesize a series of glucosidase compounds, and test all synthesized compounds for in vitro alpha-glycosidase activity, hoping to provide new ideas for the research and development of hypoglycemic drugs and health-related products.

[0004] The applicant has carried out research on the synthesis of glucosinolate compounds. During the research process, the applicant has experimentally concluded that glucosinolate compounds 1 to 5 have an inhibitory effect on alpha-glycosidase. Summary of the invention

[0005] The object of the present invention is to provide an application of aromatic glucosinolates in the preparation of hypoglycemic drugs and health-related products, characterized in that the drugs contain aromatic glucosinolate compounds.

[0006] α-Glycosidase is α-glucosidase.

[0007] The structure of aromatic glucosinolates is: Advantages of the present invention: The present invention provides the use of aromatic glucosinolates in the preparation of hypoglycemic drugs and health-related products, and the glucosinolate compounds have a good inhibitory effect on α-glucosidase. The preparation method of the glucosinolate compounds provided by the present invention is simple and is expected to further solve the problem of diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 It is the structural formula of aromatic glucosinolate compound; Figure 2 This is the synthetic route of compounds 1~2; Figure 3 This is the synthetic route of compounds 3~5. DETAILED DESCRIPTION

[0009] The present invention is further described below, but the present invention is not limited in any way. Any changes made based on the present invention belong to the protection scope of the present invention.

[0010] Provided is the use of glucosinolates in the preparation of hypoglycemic drugs and health-care related products, wherein the drugs contain glucosinolate compounds.

[0011] α-Glycosidase is α-glucosidase.

[0012] The structural formula of thioglycoside compounds is shown in Figure 1 .

[0013] Preparation of glucosinolate compounds: (1) Synthesis of p-tolyl-2, 3, 4, 6-tetra- O -Acetyl-1-thio- β -D-pyranoglucoside 1: D-glucose is used as the starting material, and after full acetylation, it is reacted with p-toluene thiophenol under the action of Lewis acid boron trifluoride ether to synthesize compound 1. The synthetic route is shown in Figure 2 .

[0014] (2) Synthesis of p-tolyl-2, 3, 4, 6-tetra- O -Benzyl-1-thio- β -D-pyranoglucoside 2: Compound 1 is used as the starting material, and the hydroxyl group is protected with BnBr to obtain compound 2. The synthetic route is shown in Figure 2 .

[0015] (3) Synthesis of p-tolyl-2- O -acetyl-3, 4, 6-tri- O -Benzyl-1-thioglycoside derivatives 3~5: Sugars of different configurations are fully acetylated and brominated at position 1. The brominated sugar is reacted in dry acetonitrile, anhydrous ethanol, TBAI and TEA under heating reflux at 60°C to obtain an orthoesterified product. The acetyl groups at positions 3, 4, and 6 are treated with BnBr, KOH, and heated under reflux at 70°C to generate an orthoesterified product protected by benzyl groups at positions 3, 4, and 6. Then p-toluene thiophenol is added and dissolved in DCM. Under the protection of N2, boron trifluoride ether is added dropwise to react to obtain compounds 3~5. The specific synthetic route is shown in Figure 3 .

[0016] Example 1 Synthesis method of compound 2

[0017] Add NaOAc (1 g, 12.2 mmol) and Ac2O (9.4 mL, 99.4 mmol) to the reaction flask, heat to reflux, add D-glucose (1.8 g, 10 mmol) in batches, and reflux and stir to react (TLC monitoring). Cool the reaction solution to room temperature, add crushed ice, and sonicate to precipitate a white solid. Filter, wash with water until there is no sour taste, dissolve the filter cake with hot EtOH, add PE, let stand and cool to precipitate a solid, filter, wash the filter cake with PE, and obtain 3.8 g of a white solid with a yield of 97%. Add a white solid (2 g, 5.1 mmol), 2 g of 4Å molecular sieves, and p-toluene thiophenol (1.3 g, 10.5 mmol) to a dry round-bottom flask, and add dry DCM (20 mL) to dissolve. Cool to 0 ℃, slowly add BF3·OEt2 (2.2 mL, 17.5mmol) under N2 protection, and after the addition is complete, warm to room temperature and stir to react overnight (TLC monitoring). The mixture was washed with saturated Na2CO3 solution, extracted with DCM, the organic layers were combined, washed with saturated NaCl solution, dried with anhydrous Na2SO4, filtered, and the filtrate was concentrated and chromatographed on a silica gel column (eluent: PE / EA=4 / 1, V / V ) was purified to obtain a white solid 1 (p-tolyl-2, 3, 4, 6-tetra- O -Acetyl-1-thio- β -D-pyranoglucoside) 1.8 g, yield 78%. mp 113.9~117.6℃.

[0018] 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 completion by TLC. Evaporate 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 at room temperature overnight, and monitor the reaction completion by TLC. Add water (5 mL) to quench the reaction, dilute the reaction solution with EA, wash with water and saturated NaCl solution, dry over anhydrous Na2SO4, filter, concentrate the filtrate, and chromatograph the residue on a silica gel column (eluent: PE / EA=35 / 1~PE / EA=14 / 1, V / V ), to obtain a white solid 2 (p-tolyl-2, 3, 4, 6-tetra- O -Benzyl-1-thio- β -D-pyranoglucoside) 1.99 g, yield 70.1%. mp 95.6~97.7 °C.

[0019] The method for synthesizing compounds 3~5 is illustrated by taking the synthesis of 3 as an example.

[0020] Add D-galactose (4 g, 22.2 mmol), Ac2O (20 mL, 211.6 mmol), I2 (180 mg, 0.71 mmol) to the reaction flask and stir at room temperature until the reaction system turns brown and transparent. After the reaction is complete, add dry DCM (100 mL), cool the reaction solution to 0 °C, drop 33% HBr-AcOH solution (42 mL), stir the reaction, remove the ice-water bath, and stir the reaction at room temperature for 1 h (TLC monitors 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, and concentrate the filtrate to obtain 7.5 g of colorless oily liquid crude product. Take the crude product (5 g, 12.2 mmol), 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), and react at 60 °C for 1 h (TLC monitoring reaction). Dilute the reaction solution with EA, wash with water and saturated NaCl solution in turn, dry with anhydrous Na2SO4, filter, concentrate the filtrate under reduced pressure, and chromatograph on a silica gel column (eluent: PE / EA=4 / 1, V / V ), 4.1 g of colorless oily liquid was obtained. A portion of it (3.6 g, 9.9 mmol) was dissolved in MeOH (36 mL), and MeONa (113 mg, 2.1 mmol) was added. The reaction was stirred at room temperature for 1 h. After the reaction was complete, the concentrated reaction solution was pumped on a diaphragm vacuum pump for 0.5 h. DMF (80 mL) was added to dissolve, and NaH (3.96 g, 99 mmol, 60%) was added at 0°C. After stirring the reaction until no bubbles were generated, BnBr (11.8 mL, 99 mmol) was added and stirred at room temperature overnight (TLC detection reaction). After the reaction was complete, MeOH (50 mL) was added to quench the reaction, and the residue was distilled under reduced pressure. It was diluted with Et2O (300 mL), 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, V / V ), a yellow oily liquid (1, 2-methyl orthoacetate-3, 4, 6-tri- O -Benzyl- α -D-galactopyranoside) 3.4 g, yield 51%.

[0021] Dissolve the yellow oily liquid (1.9 g, 3.6 mmol) in 60% AcOH (30 mL) and stir at room temperature for 4 h. After the reaction is complete, spin dry the reaction solution and dry it with toluene (10 mL) three times. Add Pyr (30 mL) to the crude product to dissolve it, and then add DMAP (88 mg, 0.7 mmol). Cool to 0 °C, slowly add Ac2O (0.7 mL, 7.2 mmol) under N2 protection, and then warm to room temperature and stir to react overnight. After the reaction is completed, cool the reaction system to 0 °C and add anhydrous ethanol (1 mL) to quench the reaction. The reaction solution was spin-dried, and EA (60 mL) was added to dissolve it. The solution 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 spin-dried. The crude product was purified to obtain 1.5 g of a yellow oily liquid with a yield of 81%.

[0022] Take the yellow oil (1.5 g, 2.9 mmol) and p-toluene thiophenol (0.7 g, 5.8 mmol) and dissolve them in dry DCM (20 mL). Place it in an ice-water bath, slowly add 48% BF3·OEt2 (0.4 mL, 4.3mmol) under N2 protection, then return to room temperature, continue stirring for 4 h, and monitor the reaction by TLC. Dilute the reaction solution with dichloromethane, wash the organic layer with saturated sodium bicarbonate solution, water and saturated NaCl solution, and dry it over anhydrous Na2SO4. Filter, and the filtrate is dried to obtain a crude product. The crude product is 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%.

[0023] p-Tolyl-2- O -acetyl-3, 4, 6-tri- O -Benzyl-1-thio- β -D-pyranoglucoside (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. 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). Example 2 Alpha-glucosidase activity test 1. Materials and Methods 1. Drugs and samples Glucoside compounds 1-5 were prepared by Example 1.

[0024] All test samples were dissolved in DMSO and stored in a refrigerator at 4°C.

[0025] 2. Experimental methods Add 112 μL potassium phosphate buffer (pH 6.8) and 20 μL 0.2U·mL -1 α-glucosidase (prepared in potassium phosphate buffer), 8 μL DMSO, 8 μL different concentrations (220.00, 110.00, 55.00, 27.50, 13.75 μmol·L -1 ) sample solution, incubated at 37°C for 15 min, and then 20 μL of 2.5 mmol·L -1 PNPG (prepared with potassium phosphate buffer), shake well, react at 37℃ for 15 min, then add 80 μL 0.2 mol·L -1 The reaction was terminated by adding Na2CO3 solution and the absorbance was measured at 405 nm. Each sample was measured three times in parallel.

[0026] Sample determination: A total of 4 groups were set up, each with 3 replicates, and the total volume of each well was 240 μL, namely: 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).

[0027] 3. Calculation formula According to I%=[1- (A c -A d ) / (A a -A b )]×100% to calculate the inhibition rate, and use Origin8.0 software to calculate the corresponding IC 50 value.

[0028] 2. Results Conclusion 1. The samples that have single drug effects on α-glucosidase are: 3, 4 In summary, glucosinolate compounds 3-4 alone have an inhibitory effect on α-glycosidase, and the inhibitory effect is good.

Claims

1. The use of aromatic glucosinolates in the preparation of α-glucosidase inhibitors, characterized in that: The structure of aromatic glucosinolates is: Among them, R1 is acetyl or benzyl, R2 is hydrogen or benzyloxy, R3 and R5 are hydrogen, acetoxy or benzyloxy, and R4 is hydrogen or acetoxy.

2. The use according to claim 1, characterized in that The structure of the aromatic glucosinolate compound is: 。 3. A hypoglycemic drug characterized by Contains the aromatic glucosinolate compound according to claim 1 or 2.

4. A blood sugar lowering health-related product, characterized in that Contains the aromatic glucosinolate compound according to claim 1 or 2.

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