Synthesis method of thioglycosyl eupatorin derivative and application of hypoglycemic drug

Abstract

The invention provides a synthesis method of a thioglycosyl eupatorin derivative. The structural formula of the compound is # imgabs0, wherein R comprises silyl ether protecting groups, ester protecting groups and ether protecting groups. The thioglycosyl eupatorin is obtained by taking eupatorin mercaptan and carbonic ester sugar as raw materials, adding a catalyst and a ligand, adding an additive and taking acetonitrile as a solvent to react under the protection of nitrogen, and directly adding potassium osmate to oxidize without separating an intermediate. The reaction product is single, high in yield and easy to separate, and has good functional group tolerance. The prepared sulfo-glycosyl eupatorin derivative has good hypoglycemic activity and is expected to be used for treating diabetes mellitus.

Description

Technical Field

[0001] The present invention provides a synthesis method of a class of thioglycosyl eupatilin derivatives and hypoglycemic drugs, belonging to the technical field of pharmaceuticals. Background Art

[0002] Carbohydrates are important components indispensable in life activities. Among them, O-glycosides are the main components of carbohydrates. Since thioglycosides are more stable than O-glycosides to chemical degradation and enzymatic hydrolysis, thioglycosides, as analogs of natural O-glycosides with biological activities, have received extensive attention and research in recent years. In addition, thioglycopeptides / proteins also show important biological activities in the life system. For example, thioglycosides such as auranofin, pirlimycin, lincomycin, and clindamycin have been developed as anti-rheumatic and antibacterial drugs. However, due to the toxicity of sulfur-containing functional groups to noble metal catalysts, the stereoselective synthesis of thioglycoside compounds is still extremely challenging at present.

[0003] Eupatilin is a benzofuran active ingredient derived from plants of the genus Eupatorium, and has various biological activities such as anti-tumor, anti-inflammatory, bactericidal, hypoglycemic, antidepressant, and antioxidant. In recent years, eupatilin and its derivatives have received extensive attention in clinical applications, new drug research and development, etc. However, at present, the acquisition of eupatilin mostly comes from chemical extraction of plants of the genus Eupatorium, and the extraction rate is not high (Research progress on the synthesis of the active compound eupatilin, Chemical Reagents, 2023, 45(6): 131-138.). In order to deeply develop and utilize the active molecule eupatilin, on the basis of the previous total synthesis of eupatilin by this research group (Open-chain polymers of eupatilin, preparation method and use ZL202211519521.9, a class of eupatilin chalcone compounds, preparation method and its application ZL202310054740.2), the synthesis method of thioglycosyl eupatilin derivatives and their hypoglycemic activities were further studied, in order to provide new candidate drugs for the treatment of diabetes. Summary of the Invention

[0004] The present patent aims to combine eupatilin with different types of monosaccharides through thioglycosidic bonds, and enhance the hypoglycemic activity of eupatilin through the pharmacological properties of thioglycosides. The present invention establishes a synthesis route with easily available raw materials, simple operation, mild conditions, short route and high yield, providing a raw material basis for the research related to thioglycosyl eupatilin derivatives.

[0005] In view of the above technical problems, the present invention provides a thioglycosyl eupatilin derivative, 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 sulfosugar eupatilin derivatives are selected from any one of the following formulas: .

[0007] The present invention also provides a method for preparing a sulfosugar eupatilin derivative, comprising the following steps: (1) Under nitrogen protection, eupatilin thiol and allyl glycosyl carbonate are mixed, then catalyst 1 is added, followed by a ligand. After evacuating the air, triethylamine and acetonitrile are added thereto, and the mixture is stirred at 60 °C for reaction. The reaction progress is monitored by TLC until the eupatilin thiol is completely consumed; without separating the intermediate, an oxidant is directly added for the subsequent reaction. After the reaction is completed as monitored by TLC, it is rapidly quenched, extracted, dehydrated, concentrated under reduced pressure, and then purified by column chromatography to obtain the sulfosugar eupatilin derivative. The reaction formula is as follows: ; 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.

[0008] In the step (1), 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 amount of the catalyst used is 5-10 mol% of the molar amount of eupatilin thiol.

[0009] In the step (1), the ligand includes any one of ([[]] R )-BINAP, ([[]] S )-BINAP, MeO-BIPHEP, (S)-SegPhos, ([[]] R )-JosiPhos, and ([[]] S )-JosiPhos, etc. The amount of the ligand used is 10-15 mol% of the molar amount of eupatilin thiol.

[0010] 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 yields.

[0011] In the step (1), the solvent is selected from one or a combination of two of acetonitrile, toluene, dichloromethane, tetrahydrofuran, and 1,4-dioxane.

[0012] In the step (1), the reaction temperature is 25~60 °C, and the reaction time is 4~6 hours.

[0013] In the step (2), the oxidant is selected from potassium osmate dihydrate, osmium tetroxide, hydrogen peroxide, tert-butyl hydroperoxide, m-chloroperbenzoic acid, etc., and the amount of the oxidant is 5~10 mol% of the molar amount of eupatilin thiol.

[0014] In the step (2), the reaction temperature is 25~60 °C, and the reaction time is 4~6 hours.

[0015] In the step (1), the solvent is selected from one or a combination of two of acetonitrile, toluene, dichloromethane, tetrahydrofuran, and 1,4-dioxane.

[0016] The present invention also provides a drug against α-glucosidase, and the drug is the thioglycosyl eupatilin derivative compound.

[0017] A drug composition against α-glucosidase, the drug composition includes the thioglycosyl eupatilin derivative compound. The composition includes excipients predictable by those skilled in the art, such as carboxymethyl cellulose, microcrystalline cellulose, mannitol, lactose, etc.

[0018] The β-configured thioglycosyl eupatilin has better hypoglycemic activity.

[0019] In the above, α-glucosidase is inhibited by hydrolyzing to produce glucose and PNP under the action of α-glucosidase.

[0020] The present invention also provides a preparation of a drug for treating diabetes, including the drug or the drug composition.

[0021] The reaction product obtained by adopting the technical scheme of the present invention is single, has a high yield, is easy to separate, and has good functional group tolerance. The thioglycosyl eupatilin derivative prepared by the present invention has good hypoglycemic activity and is expected to be used for the treatment of diabetes. Description of the Drawings

[0022] Figure 1 1H NMR spectrum of Compound 1.

[0023] Figure 2 13C NMR spectrum of Compound 1. Detailed implementation mode

[0024] Experimental reagents: Tetrakis(triphenylphosphine)palladium (analytical pure), R-BINAP (analytical pure), dichloromethane (analytical pure), petroleum ether (boiling range 60 - 90 °C, analytical pure), ethyl acetate (analytical pure), acetonitrile (analytical pure), anhydrous sodium sulfate (analytical pure), potassium osmate dihydrate (analytical pure), osmium tetroxide (analytical pure), hydrogen peroxide (analytical pure), tert-butyl hydroperoxide (analytical pure), meta-chloroperoxybenzoic acid (analytical pure), N-methylmorpholine-N-oxide (NMO, analytical pure), deuterated chloroform (deuterium atom content 99.8%, TMS content 0.03% V / V); NMR tube (5mm 100 / pk 2 ST500-8).

[0025] Experimental instruments: ZXZ-4 rotary vane vacuum pump (Linhai Tanshi Vacuum Equipment Co., Ltd.), 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 Co., Ltd.), EYELA SB-1100 rotary evaporator (Shanghai Ailang Instrument Co., Ltd.), FA2104B analytical balance (Shanghai Yueping Scientific Instrument Co., Ltd.), DF-101S heating magnetic stirrer with heat collection and constant temperature (Gongyi Yingyu Yuhua Instrument Factory), ZF-6 three-purpose ultraviolet analyzer (Shanghai Jiapeng Technology Co., Ltd.), Ultrashied 400 MHz Plus nuclear magnetic resonance instrument (Bruker, Switzerland).

[0026] Example 1 Weigh 0.1 mmol of eupatilin thiol and 0.15 mmol of glycofuranose carbonate (1,5-anhydro-6- O -( tert -butyldiphenylsilyl)-3,4- O -carbonate-2-deoxy-D-lyxo-hex-1-enopyranose, CAS No. 151265-18-8) sugar into a Schlenk flask, add 10 mol% of tetrakis(triphenylphosphine)palladium and 10% mol of R-BINAP, evacuate for 30 min, after purging with nitrogen, add 2 mL of anhydrous acetonitrile and 2.2 mmol of anhydrous triethylamine, and stir and react at 60 °C for 5 hours. Monitor the reaction process by thin-layer chromatography (TLC) until eupatilin thiol has reacted completely (monitor the reaction process by TLC).

[0027] Add potassium osmate dihydrate (0.12 mmol) and N-methylmorpholine-N-oxide (0.12 mmol) to the reaction flask, continue stirring the reaction for 3 hours. After monitoring the completion of the reaction by TLC, quickly quench the reaction with 15 mL of saturated sodium bicarbonate solution, extract twice with 15 mL of ethyl acetate. Separate the organic phase obtained, dry it with anhydrous sodium sulfate, concentrate the filtrate obtained by suction filtration under reduced pressure to obtain the crude product, and then perform column chromatography using ethyl acetate as the eluent to obtain the pure product of compound 1 with a yield of 88.8% and an HPLC purity of 99.2%. The specific reaction formula is as follows: .

[0028] Example 2 Prepared by the method of Example 1, with the ligand replaced by ( R )-Tol-BINAP, the yield is 80.2% and the HPLC purity is 98.1%.

[0029] Example 3 Prepared by the method of Example 1, with only the ligand replaced by ( R )-MeO-BIPHEP, the yield is 78.6.6% and the HPLC purity is 97.3%.

[0030] Example 4 Prepared by the method of Example 1, with only the ligand replaced by ( R )-SegPhos, the yield is 79.6% and the HPLC purity is 98.5%.

[0031] Example 5 Prepared by the method of Example 1, with only the ligand replaced by ( R )-Josiphos, the yield is 77.7% and the HPLC purity is 97.2%.

[0032] Example 6 Prepared by the method of Example 1, with only the ligand replaced by ( R , R )-Me-DuPhos, the yield is 85.5% and the HPLC purity is 98.1%.

[0033] Example 7 Prepared by the method of Example 1, with only the oxidant replaced by osmium tetroxide, the yield is 83.9% and the HPLC purity is 97.8%.

[0034] Example 8 Prepared by the method of Example 1, with only the oxidant replaced by hydrogen peroxide, the yield is 63.2% and the HPLC purity is 85.5%.

[0035] Example 9 Prepared by the method of Example 1, with only the oxidant replaced by tert-butyl hydroperoxide, the yield was 74.6% and the HPLC purity was 89.2%.

[0036] Example 8 Prepared by the method of Example 1, with only the silicon-based protected glycocarbonate replaced by ethoxy glycocarbonate (3aR,4R,7aR)-4-(ethoxymethyl)-4,7a-dihydro-3aH-[1,3]dioxolo[4, 5-c]pyran-2-one, the yield of product 4 was 90.3% and the HPLC purity was 98.6%.

[0037] Example 9 Prepared by the method of Example 1, with only the silicon-based protected glycocarbonate replaced by acetoxy glycocarbonate ((3aR,4R,7aR)-2-oxo-4,7a-dihydro-3aH-[1,3]dioxolo[4, 5-c]pyran-4-yl)methyl acetate, the yield of product 6 was 88.7% and the HPLC purity was 99.3%.

[0038] Prepared by the method of Example 1, with only the glycocarbonate replaced, the structure of the target compound was obtained: .

[0039] Spectral data: 1-(2-(2-(((2S,3R,4R,5R,6R)-6-(((tert-butyldiphenylsilyl)oxy)methyl)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)thio)propan-2-yl)-6-hydroxybenzofuran-5-yl)ethan-1-one (Compound 1):

[0040] 1 H NMR (400 MHz, Chloroform- d ) δ 12.41 (s, 1H), 7.77 (s, 1H), 7.72 –7.65 (m, 2H), 7.65 – 7.60 (m, 2H), 7.52 – 7.35 (m, 6H), 6.79 (s, 1H), 6.49(s, 1H), 4.56 (d, J= 10.3 Hz, 1H), 4.06 (t, J = 3.5 Hz, 1H), 3.95 (t, J = 3.8 Hz,1H), 3.76 – 3.65 (m, 2H), 3.62 – 3.44 (m, 2H), 3.17 (d, J = 4.2 Hz, 1H), 2.60(s, 3H), 2.42 (s, 1H), 2.38 (s, 1H), 1.80 (s, 3H), 1.78 (s, 3H), 1.01 (s,9H). 13 C NMR (100 MHz, CDCl 3 ) δ 204.0, 162.0, 161.3, 159.6, 135.9, 135.7, 132.8,132.6, 130.1, 130.0, 128.98, 127.95, 123.5, 120.9, 117.0, 102.6, 99.7, 83.2,74.6, 70.4, 69.8, 66.6, 64.8, 45.6, 28.6, 28.3, 26.9, 19.2. Evaluation of the α-glucosidase inhibitory activity of sulfoglycosyl eupatilin derivatives: Preparation of 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).

[0041] The experiment was divided into an enzyme activity group (50 μL of α-glucosidase solution and 20 μL of PBS solution), an enzyme blank group (70 μL of PBS solution), a sample group (50 μL of α-glucosidase solution and 20 μL of sample solution), a positive group (50 μL of α-glucosidase solution and 20 μL of positive drug solution (acarbose)), a sample blank group (50 μL of PBS solution and 20 μL of sample solution), and a positive blank group (50 μL of PBS solution and 20 μL of positive drug solution (acarbose)). The drugs in each group were added to 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 3Terminate the reaction of the solution and mix it by shaking 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, measure its absorbance, and the inhibition rate and IC 50 value of α-glucosidase in each sample can be calculated according to the formula.

[0042] 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 method for synthesizing a class of sulfosylzetranin derivatives, 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 sulfosaccharosylzetranin derivative according to claim 1, characterized in that The sulfosaccharide zearalan derivative is selected from any one of the following formulas: 。 3. The method for preparing the sulfosaccharosylzeuratin derivative according to claim 1, characterized in that: The steps include: Under nitrogen protection, the zeulanin thiol and the glycosyl carbonate are mixed, and then the catalyst 1 is added, and then the ligand is added. After vacuuming, triethylamine and acetonitrile are added thereto, and the reaction is stirred at 60°C. The reaction progress is monitored by TLC. After the zeulanin thiol is completely reacted, the intermediate is not separated, and the oxidant is directly added for subsequent reaction. After the reaction is completed by TLC monitoring, the reaction is quickly quenched, extracted, dehydrated, concentrated under reduced pressure, and purified by column chromatography to obtain a sulfosylzeulanin derivative.

4. The method for preparing the sulfosaccharosylzeulantrine derivative according to claim 3, characterized in that: The catalyst 1 in step (1) is selected from any one of Pd2(dba)3, Pd(OAc)2, Pd(MeCN)2Cl2, and Pd(PPh3)4; The ligand includes ( R )-BINAP, ( R )-Tol-BINAP, ( R )-MeO-BIPHEP, ( R )-SegPhos, ( R )-Josiphos or ( R , R )-Me-DuPhos; The reaction temperature is 25~60°C and the reaction time is 4~6 hours.

5. The method for preparing the sulfosaccharosylzeulin derivative according to claim 3, characterized in that: In step (2), the oxidant is selected from potassium osmate dihydrate, osmium tetroxide, hydrogen peroxide, tert-butyl peroxide or m-chloroperbenzoic acid.

6. The method for preparing the sulfosaccharosylzeulin derivative according to claim 3, characterized in that: In step (2), 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 sulfosylzetranin derivative according to claim 1 or 2.

8. An anti-α-glucosidase pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the sulfosylzetran derivative 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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