Synthesis method and hypoglycemic activity of eupatorin glycal thioglycoside

Abstract

The invention provides a synthesis method of eupatorin glycal thioglycoside compounds. The structural formula of the compounds is # imgabs0 # eupatorin glycal thioglycoside. The eupatorin glycal thioglycoside takes eupatorin mercaptan and carbonic ester sugar as raw materials, tetrakis (triphenylphosphine) palladium as a catalyst, R-BINAP as a ligand, triethylamine as an additive and methylbenzene as a solvent to react under the protection of nitrogen. The reaction product is single, high in yield and easy to separate, and has good functional group tolerance. The product prepared by the invention has good hypoglycemic activity and is expected to be used for treating diabetes mellitus.

Description

Technical Field

[0001] The present invention provides a method for synthesizing eupatilinene glycosylthioglycosides and their hypoglycemic activity, belonging to the field of pharmaceutical technology. Background Art

[0002] Eupatilin is a benzofuran compound isolated from plants of the genus Eupatorium, which has a wide range of biological activities such as anti-inflammatory, anti-tumor, antibacterial, and hypoglycemic. Literature research found that the structural modification of eupatilin may enhance its hypoglycemic activity. In recent years, most of the structural modifications of eupatilin are to add some substituents to the structural core of eupatilin to make some functional group transformations. For example, in the previous research of this research group, a chalcone structure was connected to the core structure of eupatilin, and it was found that it has good hypoglycemic activity.

[0003] Sulfur is an essential element for life and is widely present in biomedical materials, fine chemicals, and drugs. Most natural products are mainly oxygen glycosides. Glycosides formed by replacing oxygen atoms with sulfur atoms are usually used as glycosyl donors to construct various glycosidic bonds. On the other hand, thioglycosides are more resistant to enzymatic cleavage and chemical degradation and have higher metabolic stability, and have extensive applications in biological research and drug development. For example, the anti-rheumatic drug Auranofin, the antibacterial drugs Lincomycin and Clindamycin, and the drug Sotagliflozin for treating heart failure, type 2 diabetes, and chronic kidney disease all contain thioglycoside structures. Therefore, replacing oxygen glycosides with thioglycosyl analogs has become an important means to improve pharmacological activity. However, due to the strong nucleophilicity of sulfur-containing functional groups and the toxicity of catalysts, the stereoselective control of this method is often difficult. Based on the previous development of hypoglycemic drugs from eupatilin derivatives in this research group (open-chain polymers of eupatilin, preparation methods and uses ZL202211519521.9, a class of eupatilin chalcone compounds, preparation methods and their applications ZL202310054740.2), the synthesis method of eupatilinene glycosylthioglycosides and their hypoglycemic activity uses have been further studied in order to provide new candidate drugs for the treatment of diabetes. Summary of the Invention

[0004] This patent aims to combine eupatilin with sugar through a thioglycoside bond to enhance the hypoglycemic activity of eupatilin through the pharmacological properties of thioglycosides. The present invention establishes a synthetic route with easily available raw materials, simple operation, mild conditions, short route, and high yield, providing a raw material basis for the research on eupatilinene glycosylthioglycoside compounds.

[0005] In view of the above technical problems, the present invention provides a class of eupatilinene glycosylthioglycoside compounds, and the structural formula of the compound is , the R includes siloxy groups (such as triisopropylsiloxy, tert-butyldimethylsiloxy, tert-butyldiphenylsiloxy, etc.), alkoxy groups (such as methoxy, ethoxy, n-propoxy, isopropoxy, benzyloxy, etc.), and ester groups (such as formyloxy, acetoxy, tert-butoxycarbonyloxy, benzoyloxy, etc.).

[0006] In some preferred embodiments, the eupatilin glycoside thio compounds are selected from any one of the following formulas: .

[0007] The present invention also provides a method for preparing eupatilin glycoside thio compounds, which includes the following steps: Under nitrogen protection, mix eupatilin thiol and enose carbonate, add a catalyst, then add a ligand, evacuate the air, add triethylamine and toluene thereto, stir at room temperature for reaction, monitor the reaction progress by TLC. When the eupatilin thiol reacts completely, terminate the reaction, quickly quench, extract, dehydrate, concentrate under reduced pressure, and then purify by column chromatography to obtain eupatilin glycoside thio compounds. The reaction formula is as follows: .

[0008] The R includes siloxy groups, alkoxy groups, and ester groups; the siloxy groups are selected from triisopropylsiloxy, tert-butyldimethylsiloxy or tert-butyldiphenylsiloxy, the alkoxy groups are selected from methoxy, ethoxy, n-propoxy, isopropoxy or benzyloxy; the ester groups are selected from formyloxy, acetoxy, tert-butoxycarbonyloxy or benzoyloxy.

[0009] The catalyst is selected from any one of Pd 2 (dba) 3 , Pd(OAc) 2 , Pd(MeCN) 2 Cl 2 , Pd(PPh 3 ) 4 . The dosage of the catalyst is 5 - 10 mol% of the molar amount of eupatilin thiol.

[0010] The ligand includes any one of ([[]] R )-BINAP, ([[]] R )-Tol-BINAP, ([[]] R )-MeO-BIPHEP, ([[]] R )-SegPhos, ([[]] R )-Josiphos and ([[]] R , R )-Me-DuPhos, etc. The dosage of the ligand is 10 - 15 mol% of the molar amount of eupatilin thiol.

[0011] The ( R)-configured chiral ligands are conducive to coordinating with palladium to form a π-allyl complex intermediate, and are more conducive to obtaining β-configured thioglycosides in high yield.

[0012] The solvent described above is selected from one or a combination of two of toluene, acetonitrile, dichloromethane, tetrahydrofuran, and 1,4-dioxane.

[0013] The reaction temperature is 25~60 °C, and the reaction time is 4~6 hours.

[0014] The present invention also provides a drug against α-glucosidase, and the drug is the eupatilinene sugar thioglycoside compound described above.

[0015] The β-configured eupatilinene sugar thioglycoside has better hypoglycemic activity.

[0016] A drug composition against α-glucosidase, the drug composition includes the eupatilinene sugar thioglycoside compound described above. The composition includes excipients that can be predicted by those skilled in the art, such as carboxymethyl cellulose, microcrystalline cellulose, mannitol, lactose, etc.

[0017] In the above, α-glucosidase is inhibited by hydrolyzing to produce glucose and PNP under the action of α-glucosidase.

[0018] The present invention also provides a method for preparing a drug for treating diabetes, including the drug or the drug composition described above.

[0019] The reaction product obtained by adopting the technical solution of the present invention is single, has a high yield, is easy to separate, and has good functional group tolerance. The product prepared by the present invention has good hypoglycemic activity and is expected to be used for the treatment of diabetes. Description of the Drawings

[0020] Figure 1 It is the hydrogen spectrum diagram of compound 1.

[0021] Figure 2 It is the carbon spectrum diagram of compound 1. Detailed Embodiments

[0022] Experimental reagents: Palladium tris(triphenylphosphine) (analytical pure), R-BINAP (analytical pure), dichloromethane (analytical pure), petroleum ether (boiling range 60-90 °C, analytical pure), ethyl acetate (analytical pure), toluene (analytical pure), anhydrous sodium sulfate (analytical pure), deuterated chloroform (deuterium atom content 99.8%, TMS content 0.03% V / V); NMR tube (5mm 100 / pk 2 ST500-8).

[0023] Experimental instruments: ZXZ-4 Rotary Vane Vacuum Pump (Taishi Vacuum Equipment Co., Ltd., Linhai City), DZF-6020 Vacuum Drying Oven (Shanghai Xinmiao Medical Instrument Manufacturing Co., Ltd.), SHB-IIIA Circulating Water Multi-Purpose Vacuum Pump (Shanghai Yukang Scientific and Educational Instrument Equipment Co., Ltd.), CL-4 Flat Magnetic Stirrer (Zhengzhou Great Wall Science and Industry & Trade Co., Ltd.), EYELA SB-1100 Rotary Evaporator (Shanghai Ailang Instrument Co., Ltd.), FA2104B Analytical Balance (Shanghai Yueping Scientific Instrument Co., Ltd.), DF-101S Thermostatic Heating Magnetic Stirrer with Heat Collection (Yuhua Instrument Factory, Yingyu, Gongyi City), ZF-6 Three-Purpose Ultraviolet Analyzer (Shanghai Jiapeng Technology Co., Ltd.), Ultrashied 400 MHz Plus Nuclear Magnetic Resonance Instrument (Bruker, Switzerland).

[0024] Example 1 The technical route is as follows: Weigh 0.10 mmol of eupatilin thiol and 0.10 mmol of allyl carbonate (1,5-anhydro-6- O -( tert -butyldiphenylsilyl)-3,4- O -carbonate-2-deoxy-D-lyxo-hex-1-enopyranose, CAS No. 151265-18-8) into a Schlenk flask. Add 10 mol% of tetrakis(triphenylphosphine)palladium and 10% mol of ( R )-BINAP. Evacuate for 30 min, replace with nitrogen, then add 2 mL of anhydrous toluene to dissolve. Add 2.2 mmol of anhydrous triethylamine, and stir the reaction at 60 °C for 5 hours. Monitor the reaction progress by thin-layer chromatography (TLC) until the eupatilin thiol is completely reacted. Add 15 mL of saturated sodium bicarbonate solution to quickly quench the reaction. Extract twice with 15 mL of ethyl acetate. Dry the separated organic phase with anhydrous sodium sulfate, filter by suction, and concentrate the filtrate under reduced pressure to obtain the crude product. Then, use a petroleum ether / ethyl acetate solution as the eluent for column chromatography to obtain the pure product of Compound 1, with a yield of 86.5% and an HPLC purity of 98.6%. The specific reaction formula is as follows: .

[0025] Example 2 Prepare using the method of Example 1, and replace the ligand with ( R )-Tol-BINAP, with a yield of 78.5% and an HPLC purity of 97.2%.

[0026] Example 3 Prepare using the method of Example 1, and only replace the ligand with ( R)-MeO-BIPHEP, yield 79.6%, HPLC purity is 96.9%.

[0027] Example 4 Prepared by the method of Example 1, only the ligand was replaced with ( R )-SegPhos, yield 75.3%, HPLC purity is 97.1%.

[0028] Example 5 Prepared by the method of Example 1, only the ligand was replaced with ( R )-Josiphos, yield 73.1%, HPLC purity is 95.9%.

[0029] Example 6 Prepared by the method of Example 1, only the ligand was replaced with ( R , R )-Me-DuPhos, yield 82.7%, HPLC purity is 97.9%.

[0030] Example 7 Prepared by the method of Example 1, only the glycocarbonate was replaced with fucosyl glycocarbonate 2,6-anhydro-3,4- O -carbonate-1,5-dideoxy-L-arabino-hex-5-enitol, the yield of product 10 was 89.4%, HPLC purity is 98.3%.

[0031] Example 8 Prepared by the method of Example 1, only the glycocarbonate was replaced with arabinosyl glycocarbonate 1,5-anhydro-3,4- O -carbonate-2-deoxy-L-erythro-pent-1-enitol, the yield of product 11 was 94.3%, HPLC purity is 97.7%.

[0032] Prepared by the method of Example 1, only replacing the glycose, the product with the following structural formula can be obtained: Structure of the target compound

[0033] Note: For compound 12, glucose has a locked ring at the 4,6 positions of p-methoxybenzylidene (PMP), and the rest have a carbonate locked ring at the 3,4 positions.

[0034] Spectral data 1-(2-(2-((( 2S , 5R , 6R)-6-((( tert -Butyldiphenylsilyl)oxy)methyl)-5-hydroxy-5,6-dihydro-2H-pyran-2-yl)thio)propan-2-yl)-6-hydroxybenzofuran-5-yl)ethan-1-one (Compound 1): 1 H NMR (400 MHz, Chloroform- d ) δ 12.43 (s, 1H), 7.80 (s,1H), 7.67 (dt, J = 7.9, 1.7 Hz, 4H), 7.48 – 7.33 (m, 6H), 6.98 (s, 1H), 6.51(s, 1H), 6.05 (ddd, J = 10.0, 5.6, 2.3 Hz, 1H), 5.77 (dd, J = 10.0, 1.6 Hz, 1H),5.10 – 4.99 (m, 1H), 3.90 – 3.83 (m, 1H), 3.79 (dd, J = 10.5, 6.1 Hz, 1H), 3.67(dd, J = 10.5, 6.4 Hz, 1H), 3.58 (td, J = 6.2, 1.9 Hz, 1H), 2.61 (s, 3H), 1.86(s, 3H), 1.75 (s, 3H), 1.73 (s, 1H), 1.07 (s, 9H). 13 C NMR (100 MHz, CDCl 3 ) δ 203.8, 161.9, 161.2, 159.6, 135.6, 135.5, 133.5, 133.3, 130.5, 129.80,129.75, 129.7, 127.74, 127.73, 123.3, 120.9, 116.9, 102.8, 99.8, 79.8, 78.8,63.3, 61.7, 45.9, 29.1, 28.0, 26.9, 26.8, 19.3. Evaluation of the α-glucosidase inhibitory activity of eupatorin glycoside sulfur compounds: Prepare the reaction solution: PBS solution (0.1 mM, pH 6.8), solution of the target compound (4 mM), α-glucosidase solution (0.2 U / mL), Na 2 CO 3 solution (0.2 mol / L).

[0035] The experiments were divided into an enzyme activity group (50 μL α-glucosidase solution and 20 μL PBS solution), an enzyme blank group (70 μL PBS solution), a sample group (50 μL α-glucosidase solution and 20 μL sample solution), a positive group (50 μL α-glucosidase solution and 20 μL positive drug solution (acarbose)), a sample blank group (50 μL PBS solution and 20 μL sample solution), and a positive blank group (50 μL PBS solution and 20 μL positive drug solution (acarbose)). The drugs of each group were added into a 96-well plate according to the above doses. There were three parallels in each group. After shaking and mixing for 2 min, the reaction was carried out at 37 °C for 10 min. Then 25 μL of PNPG solution was added, and after shaking and mixing for 2 min, the reaction was carried out at 37 °C for 30 min. Then 100 μL of Na 2 CO 3 solution was added to terminate the reaction, and after shaking and mixing for 5 min. Since PNPG can be hydrolyzed to produce glucose and PNP under the action of α-glucosidase, and PNP has the maximum absorption at 405 nm, its absorbance was measured. According to the formula, the inhibition rate of α-glucosidase and the IC 50 value of each sample can be calculated.

[0036] Calculation formula: Inhibition rate = (1 - △ 样 / △ 酶 ) × 100% △ 样 = Average OD value of the sample - OD value of the sample blank △ 酶 = Average OD value of the enzyme activity - OD value of the enzyme blank .

Claims

1. A zeuratin glycoside compound, characterized in that: The structural formula of the compound is , the R includes a siloxy group, an alkoxy group, and an ester group; The siloxy group is selected from triisopropylsiloxy, tert-butyldimethylsiloxy or tert-butyldiphenylsiloxy, the alkoxy group is selected from methoxy, ethoxy, n-propoxy, isopropoxy or benzyloxy; the ester group is selected from formyloxy, acetoxy, tert-butoxycarbonyloxy or benzoyloxy.

2. The zearalenol glucosinolate compound according to claim 1, characterized in that: The zearalenone glycoside compound is selected from any one of the following formulas: 。 3. The method for preparing the zearalenol glycoside compound according to claim 1, characterized in that: After the zeuraithiol and the glycoside are mixed, a catalyst is added, and then a ligand is added. After evacuation, nitrogen is filled for protection. Triethylamine and toluene are added thereto. The reaction is stirred at room temperature for reaction. The progress of the reaction is monitored by TLC. After the zeuraithiol is completely reacted, the reaction is terminated. After rapid quenching, extraction, dehydration, and reduced pressure concentration, the zeuraithiol glycoside compound is purified by column chromatography.

4. The method for preparing the zearalenol glycoside compound according to claim 3, characterized in that: The catalyst is selected from any one of Pd2(dba)3, Pd(OAc)2, Pd(MeCN)2Cl2, and Pd(PPh3)4.

5. The method for preparing the zearalenol glycoside compound according to claim 3, characterized in that: The ligand includes ( R )-BINAP, ( R )-Tol-BINAP, ( R )-MeO-BIPHEP, ( R )-SegPhos, ( R )-Josiphos or ( R , R )-Me-DuPhos.

6. The method for preparing the zearalenol glycoside compound according to claim 3, characterized in that: The reaction temperature is 25~60°C and the reaction time is 4~6 hours.

7. An anti-α-glucosidase drug, characterized in that: The drug is the zearalenol glycoside compound according to claim 1 or 2.

8. An anti-α-glucosidase pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the zearalenol glycoside compound according to claim 1 or 2.

9. The drug according to claim 7 or the pharmaceutical composition according to claim 8, characterized in that: Under the action of α-glucosidase, it is hydrolyzed to produce glucose and PNP to inhibit α-glucosidase.

10. A drug for treating diabetes, characterized in that: The invention comprises the medicine or pharmaceutical composition according to any one of claims 7 to 9.

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