Triterpenoid phenolic acid ester derivative as well as preparation method and application thereof

Abstract

The invention relates to a triterpenoid phenolic ester derivative as well as a preparation method and application thereof, and belongs to the field of medicinal chemistry. The main structure R of the triterpene phenolic acid ester derivative is cucurbitane type triterpene compound residues, at least one # imgabs 0 # in the cucurbitane type triterpene compound residues is connected with a phenolic acid group, and the other # imgabs 1 # are independently connected with hydroxyl, methyl or methoxyl; in the phenolic acid group, X is 0, a C1-C3 saturated alkyl group or a C1-C3 unsaturated alkyl group, and R1, R2 and R3 are respectively and independently selected from hydrogen, a hydroxyl group or a methoxyl group. The triterpenoid phenolic acid ester derivative disclosed by the invention has a remarkable glycolipid metabolism regulation effect and can be used for preventing or treating occurrence and development of glycolipid metabolism disorder diseases.

Description

Technical Field

[0001] The invention relates to a triterpenoid phenolic acid ester derivative and a preparation method and application thereof, and belongs to the field of pharmaceutical chemistry. Background Art

[0002] Large-scale epidemiological surveys, clinical data analysis, molecular genetic research, and large animal experimental evidence close to human disease models show that metabolic syndrome, with glucose and lipid metabolism disorders as the core manifestation, is the biggest risk factor for diseases such as hyperlipidemia, fatty liver, diabetes, hypertension, atherosclerotic cardiovascular and cerebrovascular diseases, etc. Glucose and lipid metabolism disorders are widespread, pathogenic, and prone to occur.

[0003] The balance of sugar and lipid metabolism plays a vital role in maintaining the basic life activities of cells or organisms. The disruption of sugar and lipid homeostasis will lead to the occurrence of various metabolic diseases, such as obesity, diabetes, fatty liver, and cardiovascular disease. Studies have shown that polyphenols regulate sugar metabolism by regulating enzyme activity, stimulating insulin secretion, regulating insulin sensitivity, enhancing glucose uptake in tissues, and secretion of intestinal hormones. In addition, some polyphenols have been shown to regulate the expression of lipid metabolism-related genes, reduce blood total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) levels, and inhibit intestinal fat absorption.

[0004] Triterpenoids have a wide range of physiological activities, and some triterpenoids have been found to have significant effects on regulating glucose and lipid metabolism. More and more studies and data show that polyphenols can significantly enhance the antioxidant activity of drug molecules and increase drug efficacy after being combined with certain drug molecules by chemical means. Therefore, it is necessary to develop a conjugate of polyphenols and triterpenoids and their derivatives as a lead compound for developing glucose and lipid metabolism. Summary of the invention

[0005] In view of the technical problems existing in the prior art in preparing drugs for treating glucose and lipid metabolism diseases, the present invention provides a triterpenoid phenolic acid ester derivative and a preparation method and application thereof. The triterpenoid phenolic acid ester derivative provided by the present invention can be used as a lead compound for developing new, highly efficient and low-toxic glucose and lipid metabolism regulating compounds, and provides a synthetic template for developing glucose and lipid metabolism regulating drugs.

[0006] A triterpene phenolic acid ester derivative, characterized in that: the main structure R of the triterpene phenolic acid ester derivative is a cucurbitane-type triterpene compound residue, and the specific structural formula is as follows:

[0007]

[0008]

[0009] Among them, at least one Connecting phenolic acid groups, other Each independently connected to a hydroxyl group, a methyl group or a methoxy group; the phenolic acid group has the following structure:

[0010]

[0011] in,

[0012] X is 0, a C1-C3 saturated alkyl group or a C1-C3 unsaturated alkyl group;

[0013] R1, R2, and R3 are each independently selected from hydrogen, hydroxyl, or methoxy.

[0014] Furthermore, at least one of the cucurbitane-type triterpene compound residues Linking phenolic acid groups means that the phenolic acid groups may be monosubstituted, non-positionally polysubstituted or fully substituted.

[0015] Furthermore, other Each independently connected hydroxyl, methyl or methoxy means other When no phenolic acid group is attached, each can be independently selected from hydroxyl, methyl or methoxy.

[0016] Furthermore, the phenolic acid group and cucurbitane-type triterpenoid residues connected.

[0017] Preferably, X in the phenolic acid group is 0, or X is -C=C-.

[0018] Furthermore, the phenolic acid group structure in which X is 0 is

[0019]

[0020] Preferably, the triterpene phenolic acid ester derivative has the structural formula as shown below:

[0021]

[0022]

[0023] Another object of the present invention is to provide a method for preparing the above triterpenoid phenolic acid ester derivatives.

[0024] The triterpene phenolic acid ester derivative of the present invention can be prepared by the following method: at a temperature of 10 to 150° C., phenolic acid (A) and triterpene compound (B) are subjected to an esterification reaction in a solvent containing an esterification catalyst to obtain a total product, which is then filtered, the filtrate is concentrated and then dissolved in an organic solvent, extracted with water, and the organic phase is subjected to silica gel column chromatography or preparative liquid chromatography to obtain a triterpene phenolic acid ester derivative (Ia).

[0025]

[0026] In the above technical solution, the molar ratio of the phenolic acid to the triterpene compound is 1:1-3.

[0027] In the above technical solution, the organic solvent is one or more of dichloromethane, ethyl acetate or acetone.

[0028] In the above technical solution, the solvent system of the silica gel column chromatography is a combination of any two or three of dichloromethane, ethyl acetate, acetone, methanol and water.

[0029] In the above technical solution, the preparative liquid chromatography separation is performed by using methanol and 0.5% formic acid aqueous solution in a volume ratio of 60:40, or by using acetonitrile and 0.5% formic acid aqueous solution in a volume ratio of 70:30.

[0030] Furthermore, the triterpene phenolic acid ester derivative of the present invention can be prepared by the following method: at a temperature of 10 to 150° C., phenolic acid and acetic anhydride are reacted in a solvent containing an acid or an esterification catalyst to generate acetylated phenolic acid substituted with a hydroxyl group, and then the acetylated phenolic acid and a triterpene compound are subjected to an esterification reaction in a solvent containing an esterification catalyst to obtain a triterpene phenolic acid ester substituted with an acetoxy group, and hydrazine hydrate is added to carry out a deprotection reaction to obtain a triterpene phenolic acid ester derivative.

[0031] In the above technical solution, the molar ratio of the phenolic acid to the acetic anhydride is 1:1-3.

[0032] In the above technical solution, the molar ratio of the acetylated phenolic acid to the triterpene compound is 1:3-5.

[0033] In the above technical scheme, the methods for esterification reaction include: direct esterification method using N,N'-dicyclohexylcarbodiimide (DCC) or 4-dimethylaminopyridine (DMAP), acetylation protection-acyl chloride-esterification-deacetylation method, benzyl protection-acyl chloride or DCC esterification-hydrogenation debenzylation method.

[0034] Another object of the present invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of triterpenoid phenolic acid ester derivatives.

[0035] Another object of the present invention is to provide the use of the triterpenoid phenolic acid ester derivative or the pharmaceutical composition obtained above in the preparation of a drug having the effect of regulating glucose and lipid metabolism.

[0036] Beneficial effects of the invention: Based on the natural regulating glucose and lipid metabolism effects of polyphenols and triterpenoids, the invention combines the two through an ester bond, performs a structure-activity relationship analysis of medicinal chemistry, and fully explores potential target compounds. The triterpenoid phenolic acid ester derivatives provided by the invention have the advantages of high efficiency and low toxicity in regulating glucose and lipid metabolism, and at the same time provide a development template for studying candidate drugs with the effect of regulating glucose and lipid metabolism. DETAILED DESCRIPTION

[0037] The following non-limiting embodiments may enable a person skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way.

[0038] The test methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials described are commercially available unless otherwise specified.

[0039] Example 1

[0040] The preparation method of compound I-1 (3-O-galloyl-23E-5β, 19-epoxy-cucurbita-6, 23-dien-25-ol) comprises the following steps:

[0041] (1) Preparation of 3,4,5-triacetyl gallic acid

[0042] Add 26 mol of acetic anhydride and concentrated sulfuric acid (2% catalytic amount) to a 1000 mL three-necked flask equipped with electric stirring, water bath cooling, a reflux condenser protected by a drying tube, and a thermometer, and add 0.5 mol of gallic acid in batches under stirring. The temperature of the reaction system does not exceed 60°C. After the addition is completed, continue stirring for 10 to 20 minutes. The reaction system is heated at 90°C for 10 hours, and samples are taken for thin layer chromatography to track the progress of the reaction. Cool, filter, and vacuum dry to obtain 0.46 mol of 3,4,5-triacetyl gallic acid, with a yield of 92%, as a white solid.

[0043] (2) Preparation of Compound I-1

[0044] Take 0.01 mol of the above-obtained 3,4,5-triacetyl gallic acid in a 250 mL conical flask, add 50 mL of acetonitrile to dissolve it, add 0.01 mol of 23E-5β,19-epoxy-cucurbita-6,23-dien-25-ol (structure shown below) and 0.01 mol of DCC, and reflux for 24 hours; after the reaction, cool and filter the insoluble N,N'-dicyclohexylurea (DCU) white by-product, add 0.03 mol of hydrazine hydrate to the filtrate, react at room temperature for half an hour, add 0.03 mol of acetic acid and 100 mL of water, extract three times with 100 mL of ethyl acetate, combine the organic phases, wash with water until neutral, and then dry with anhydrous magnesium sulfate, and evaporate the solvent under reduced pressure to obtain the total product. The total product was separated on 200-300 mesh silica gel and gradient eluted with petroleum ether-ethyl acetate (volume ratio of 100:0, 50:1, 30:1, 10:1, 8:1, 5:1, 3:1, 2:1, 1:1, 0:1), each gradient elution was 5 column volumes, and the silica gel thin layer was used for qualitative analysis. The same components were combined and collected by elution with petroleum ether-ethyl acetate in a volume ratio of 2:1. Compound I-1 was obtained by recrystallization and vacuum drying to obtain 0.0029 mol, with a yield of 29%.

[0045]

[0046] 23E-5β,19-epoxy-cucurbita-6,23-dien-25-ol

[0047] Example 2

[0048] The preparation method of compound I-2 (3-O-dicaffeoyl-23E-5β,19-epoxy-cucurbita-6,23,25-triene-3β-ol) comprises the following steps:

[0049] (1) Preparation of 3,4-diacetylcaffeic acid

[0050] Add 26 mol of acetic anhydride and concentrated sulfuric acid (2% catalytic amount) to a 1000 mL three-necked flask equipped with electric stirring, water bath cooling, a reflux condenser protected by a drying tube, and a thermometer, and add 0.5 mol of caffeic acid in batches under stirring. The temperature of the reaction system does not exceed 60°C. After the addition is completed, continue stirring for 10 to 20 minutes. The reaction system is heated at 90°C for 10 hours, and samples are taken for thin layer chromatography to track the progress of the reaction. Cool, filter, and vacuum dry to obtain 0.44 mol of 3,4-diacetyl caffeic acid, with a yield of 88%, as a white solid.

[0051] (2) Preparation of Compound I-2

[0052] Take 0.01 mol of 3,4-diacetyl caffeic acid in a 250 mL conical flask, add 50 mL of acetonitrile to dissolve it, add 0.01 mol of 23E-5β,19-epoxy-cucurbita-6,23,25-triene-3β-ol (structure shown below) and 0.01 mol of DCC, and reflux for 24 hours; after the reaction, cool and filter the insoluble N,N'-dicyclohexylurea (DCU) white by-product, add 0.03 mol of hydrazine hydrate to the filtrate, react at room temperature for half an hour, add 0.03 mol of acetic acid and 100 mL of water, extract three times with 100 mL of ethyl acetate, combine the organic phases, wash with water until neutral, and then dry with anhydrous magnesium sulfate, and evaporate the solvent under reduced pressure to obtain the total product. The total product was separated on 200-300 mesh silica gel, and gradient eluted with petroleum ether-ethyl acetate (volume ratio of 100:1, 50:1, 10:1, 8:1, 5:1, 1:1, 0:1), each gradient elution was 5 column volumes, and the silica gel thin layer was qualitatively analyzed, and the same components were combined, and the ethyl acetate elution was collected and recrystallized to obtain compound I-2. 0.0017 mol was obtained by vacuum drying, and the yield was 17%.

[0053]

[0054] 23E-5β,19-epoxy-cucurbita-6,23,25-triene-3β-ol

[0055] Example 3

[0056] The preparation method of compound I-3 (3-O-galloyl-3β,7β,25-trihydroxy-cucurbita-5,(23E)-diene-19-ol) comprises the following steps:

[0057] 0.01 mol of 3,4,5-triacetyl gallic acid obtained in Example 1 was taken into a 250 mL conical flask, 50 mL of acetonitrile was added to dissolve it, 0.01 mol of 3β,7β,25-trihydroxy-cucurbita-5,(23E)-diene-19-ol (structure shown below) and 0.01 mol of DCC were added, and refluxed for 24 hours; after the reaction, the insoluble N,N'-dicyclohexylurea (DCU) white by-product was cooled and filtered, 0.03 mol of hydrazine hydrate was added to the filtrate, reacted at room temperature for half an hour, 0.03 mol of acetic acid and 100 mL of water were added, and extracted three times with 100 mL of ethyl acetate, the organic phases were combined, washed with water until neutral, and then dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure to obtain the total product. The total product was separated on 200-300 mesh silica gel, and gradient eluted with petroleum ether-acetone (volume ratio of 100:1, 50:1, 10:1, 8:1, 5:1, 3:1, 1:1, 0:1), each gradient elution was 5 column volumes, and the silica gel thin layer was qualitative, and the same components were combined, and the collected parts were eluted with petroleum ether-acetone 3:1, and recrystallized to obtain compound I-3. 0.0016 mol was obtained by vacuum drying, and the yield was 16%.

[0058]

[0059] 3β,7β,25-trihydroxy-cucurbita-5,(23E)-diene-19-ol

[0060] Example 4

[0061] The preparation method of compound I-4 (3-O-feruloyl-charantadiol A) comprises the following steps:

[0062] (1) Preparation of 4-acetylferulic acid

[0063] Add 26 mol of acetic anhydride and concentrated sulfuric acid (2% catalytic amount) to a 1000 mL three-necked flask equipped with electric stirring, water bath cooling, a reflux condenser protected by a drying tube, and a thermometer, and add 0.5 mol of ferulic acid in batches under stirring. The temperature of the reaction system does not exceed 60°C. After the addition is completed, continue stirring for 10 to 20 minutes. The reaction system is placed at 90°C for heating and reaction for 10 hours. Samples are taken and the reaction progress is tracked by thin layer detection. Cool, filter, and vacuum dry to obtain 0.43 mol of 4-acetylferulic acid, with a yield of 86%, as a white solid.

[0064] (2) Preparation of Compound I-4

[0065] Take 0.01 mol of 4-acetylferulic acid in a 250 mL conical flask, add 50 mL of acetonitrile to dissolve it, add 0.01 mol of charantadiol A (structure shown below) and 0.01 mol of DCC, and reflux for 24 hours; after the reaction, cool and filter the insoluble N, N'-dicyclohexylurea (DCU) white by-product, add 0.03 mol of hydrazine hydrate to the filtrate, react at room temperature for half an hour, add 0.03 mol of acetic acid and 100 mL of water, extract three times with 100 mL of ethyl acetate, combine the organic phases, wash with water until neutral, and then dry with anhydrous magnesium sulfate, evaporate the solvent under reduced pressure to obtain the total product. The total product was separated on 200-300 mesh silica gel, and eluted with chloroform-methanol (volume ratio of 100:0, 50:1, 30:1, 10:1, 5:1, 3:1, 1:1, 0:1) gradient, each gradient elution was 5 column volumes, and the silica gel thin layer was qualitative, and the same components were combined, and the collected parts were eluted with chloroform-methanol volume ratio of 10:1, and recrystallized to obtain compound I-4. 0.0019 mol was obtained by vacuum drying, and the yield was 19%.

[0066]

[0067] charantadiol A

[0068] Test on hypoglycemic activity of triterpenoid phenolic acid ester derivatives obtained in Examples 1 to 4:

[0069] (I) Experimental study on the inhibition of α-glucosidase activity of the triterpenoid phenolic acid ester derivatives obtained in Examples 1 to 4, the prototype triterpenoid compounds and the prototype phenolic acid

[0070] 1. Experimental Materials and Reagents

[0071] MultiskanMK3 (microplate reader, Thermo Electron, USA); α-glucosidase (Sigma, USA); p-nitrophenyl-α-D-pyranoglucopyranoside (PNPG, purity 99%) was purchased from Sigma, USA; acarbose (trade name: Baytangping) was purchased from Bayer, Germany; dimethyl sulfoxide (DMSO) was purchased from Laiyang Chemical Experimental Plant; other chemical reagents used were commercially available analytical grade and purchased from Sinopharm Group.

[0072] 2. Reagent preparation

[0073] a. Preparation of phosphate buffer: Na2HPO4·3H2O (35.82 g) + NaH2PO4 (15.61 g) as solute, dissolved in double distilled water, and fixed to volume in a 500 mL volumetric flask. At this time, the solution is slightly acidic, and NaOH (2 mol / L) solution is used to adjust the pH to 6.8. Store away from light.

[0074] b. Preparation of α-glucosidase solution: Weigh an appropriate amount and dissolve it in double distilled water to ensure that the activity unit of the enzyme solution is 0.02U / μL. After preparation, freeze it at -4℃ and store it away from light as a mother solution.

[0075] c. Preparation of substrate solution: Weigh 30 mg of solid 4-nitrophenyl-β-D-pyranoglucoside (PNPG) and dissolve it in 5 mL of phosphate buffer (6 μg / μL). The substrate is difficult to dissolve and requires ultrasonic treatment for about 20 minutes. After dissolution, wrap it in tin foil and store it away from light.

[0076] d. Preparation of Na2CO3 stop solution: Dissolve 21.2g Na2CO3 in distilled water to make a 0.2mol / L solution.

[0077] e. Preparation and preparation of test solution samples: Triterpenoid phenolic acid ester derivatives are all white powders, which are dissolved in DMSO to prepare mother solutions and stored in a -20℃ refrigerator. Dilute them with the corresponding culture medium before use for the experiment. The final concentration of DMSO is ≤1‰.

[0078] 3. Experimental methods

[0079] Add 30 μL of α-glucosidase solution and 20 μL of prepared triterpenoid phenolic acid ester derivatives to a test tube, incubate at 37°C for 5 minutes, add 150 μL of PNPG and 800 μL of phosphate buffered saline. Seal and incubate at 37°C for 30 minutes. Add 2 mL of 2M Na2CO3 to terminate the reaction. Measure the OD value at a wavelength of 405 nm using an ELISA reader. Use acarbose as a positive control. The inhibition rate is calculated according to the following formula, where A 空白 is the absorbance value after reaction without adding sample, A 样品 is the absorbance value after adding sample.

[0080]

[0081] The inhibition rate corresponding to each concentration is the ordinate Y, and the logarithm of the drug concentration is the abscissa X. Linear regression is performed, and the drug concentration when the inhibition rate is 50% is calculated according to the obtained equation, that is, the IC of the drug is obtained. 50 .

[0082] The concentrations of compound I-1 were 10, 3, 0.3, and 0.03 μM, respectively; the concentrations of 23E-5β,19-epoxy-cucurbita-6,23-dien-25-ol were 50, 10, 1, and 0.1 μM, respectively; the concentrations of gallic acid were 100, 50, 10, and 1 μM, respectively; the concentrations of positive drug acarbose were 30, 10, 3, and 0.3 μM. The activity experiment of inhibiting α-glucosidase was carried out according to the above method. The activity experiment of compounds I-2 to I-4 to inhibit α-glucosidase was similar to the above method, except that the types of triterpenoid compounds and phenolic acids were different. The results are shown in Table 1.

[0083] Table 1. Study on the inhibition of α-glucosidase activity by compounds I-1 to I-4

[0084]

[0085]

[0086] The study on α-glucosidase activity showed that triterpenoid phenolic acid ester derivatives had extremely strong inhibitory activity on α-glucosidase, which was stronger than the prototype triterpenoid compounds, phenolic acid and the positive control drug acarbose.

[0087] (II) Experimental study on the inhibitory activity of triterpenoid phenolic acid ester derivatives, prototype triterpenoid compounds and prototype phenolic acid obtained in Examples 1 to 4 on PTP1B

[0088] 1. Experimental materials, reagents and instruments

[0089] Nichipet EX pipette (Nichiryo); HH-B11 electric constant temperature incubator (Shanghai Yuejin Medical Instrument Co., Ltd.); HC-3518 low-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); BS-124S ​​precision electronic balance (Sartorius, Germany); 450 enzyme label quantitative tester (Bio-Rad, USA); EDTA, Tris, HCl, β-mercaptoethanol, NaOH, Na3VO4·12H2O (Sinopharm Chemical Reagent Co., Ltd.); PTP1B enzyme, DTT, pNPP (Sigma, USA); dimethyl sulfoxide (DMSO) Laiyang Chemical Experiment Plant.

[0090] 2. Reagent preparation

[0091] a. Storage solution: Take 0.6055g Tris and 0.0584g EDTA respectively, dilute to 20mL with distilled water, and adjust pH to 7.5 with concentrated hydrochloric acid. The final concentration of Tris-HCl is 250mM, and EDTA is 10mM.

[0092] b. Buffer: Take 1 mL of the stock solution, dilute it to 10 mL with distilled water, and add 0.0015 g of dithiothreitol (DTT) and 0.718 μL of β-mercaptoethanol. The buffer now contains 25 mM Tris-HCl, 1 mM EDTA, 1 mM DTT, and 2 mM β-mercaptoethanol. This solution must be prepared before use.

[0093] c. Substrate solution: Weigh 0.1856 g of 4-nitrophenyl phosphate disodium salt hexahydrate (pNPP) and dilute with 1 mL of buffer solution, dispense into 1.5 mL EP tubes, and store in a freezer at -20°C away from light.

[0094] d. Sodium vanadate solution: Take 0.816g of sodium vanadate dodecahydrate, add 100mL of distilled water, heat the solution to boil until it is translucent, and adjust the pH to about 10. Use a pipette to take 1mL of the solution into a 100mL beaker, stir with a glass rod, and heat again until it boils until it is translucent, ensuring that the sodium vanadate is in monomeric form. After cooling, divide it into 1.5mL EP tubes and store it in a freezer at -20℃.

[0095] e. Stop solution: 2M NaOH solution.

[0096] 3. Experimental methods

[0097] The triterpenoid phenolic acid ester derivatives were prepared into a 10 mM stock solution with DMSO and stored in the dark. Before use, they were diluted with PBS for experiments. The final concentration of DMSO in the highest dose group was 1‰.

[0098] Positive control group: The preparation method of sodium vanadate solution is the same as above.

[0099] The experiment of triterpenoid phenolic acid ester derivatives inhibiting PTP1B enzyme activity was carried out in a 96-well plate. The experimental group added 83 μL of enzyme buffer solution (0.4 μL enzyme), 10 μL of derivative solution and 4 μL of substrate solution in sequence; the positive control group added 83 μL of enzyme buffer solution (0.4 μL enzyme), 10 μL of sodium vanadate solution and 4 μL of substrate solution in sequence; the blank control group added 93 μL of enzyme buffer solution (0.4 μL enzyme) and 4 μL of substrate solution in sequence; the reagent control group added 83 μL of enzyme buffer solution (0.4 μL enzyme), 10 μL of DMSO solution and 4 μL of substrate solution, reacted in a constant temperature incubator at 37 ° C for 30 minutes, and then terminated the reaction with 5 μL of NaOH solution. The absorption intensity of the product at a wavelength of 405 nm was measured by an ELISA instrument, and the relative inhibition rate of the derivatives on PTP1B at different concentrations was calculated. The inhibition rate was calculated according to the following formula, where A 空白 is the absorbance value after reaction without adding sample, A 样品 is the absorbance value after adding sample.

[0100]

[0101] The inhibition rate corresponding to each concentration is the ordinate Y, and the logarithm of the drug concentration is the abscissa X. Linear regression is performed, and the drug concentration when the inhibition rate is 50% is calculated according to the obtained equation, that is, the IC of the drug is obtained. 50 .

[0102] The concentrations of compound I-1 were 10, 3, 0.3, and 0.03 μM, respectively; the concentrations of 23E-5β, 19-epoxy-cucurbita-6, 23-dien-25-ol were 50, 10, 1, and 0.1 μM, respectively; the concentrations of gallic acid were 100, 50, 10, and 1 μM, respectively; and the concentrations of positive drug sodium vanadate (Na3VO4) were 50, 25, 10, and 1 μM. The activity experiments of compounds I-2 to I-4 in inhibiting PTP1B were similar to the above method, except that the types of triterpenoid compounds and phenolic acids were different. The activity experiments of inhibiting PTP1B were carried out according to the above method, and the results are shown in Table 2.

[0103] Table 2. Study on the inhibition of PTP1B activity by compounds I-1 to I-4

[0104]

[0105] The study on PTP1B activity showed that triterpenoid phenolic acid ester compounds have extremely strong inhibitory activity on PTP1B, which is stronger than the prototype triterpenoids, phenolic acids and the positive control drug sodium vanadate.

Claims

1. A triterpenoid phenolic acid ester derivative, characterized in that: The main structure R of the triterpene phenolic acid ester derivative is a cucurbitane-type triterpene compound residue, and the specific structural formula is as follows: Among them, at least one Connecting phenolic acid groups, other Each independently connected to a hydroxyl group, a methyl group or a methoxy group; the phenolic acid group has the following structure: in, X is 0, a C1-C3 saturated alkyl group or a C1-C3 unsaturated alkyl group; R1, R2, and R3 are each independently selected from hydrogen, hydroxyl, or methoxy.

2. The triterpenoid phenolic acid ester derivative according to claim 1, characterized in that: Said X is 0, or X is -C=C-.

3. The triterpenoid phenolic acid ester derivative according to claim 1, characterized in that: The structural formula of the triterpenoid phenolic acid ester derivative is as follows:

4. The method for preparing the triterpene phenolic acid ester derivative according to any one of claims 1 to 3, characterized in that: The preparation method of the derivative comprises the following steps: at a temperature of 10 to 150° C., esterifying a phenolic acid (A) and a triterpene compound (B) in a solvent containing an esterification catalyst to obtain a total product, filtering, concentrating the filtrate and then dissolving it in an organic solvent, adding water for extraction, and subjecting the organic phase to silica gel column chromatography or preparative liquid chromatography to obtain a triterpene phenolic acid ester derivative (Ia).

5. The method for preparing the triterpene phenolic acid ester derivative according to any one of claims 1 to 3, characterized in that: The preparation method of the derivative comprises the following steps: at a temperature of 10 to 150° C., reacting phenolic acid with acetic anhydride in a solvent containing an acid or an esterification catalyst to generate acetylated phenolic acid substituted with a hydroxyl group, then performing an esterification reaction with a triterpene compound in a solvent containing an esterification catalyst to obtain triterpene phenolic acid ester substituted with an acetoxy group, and adding hydrazine hydrate to perform a deprotection reaction to obtain the triterpene phenolic acid ester derivative.

6. The preparation method according to claim 4 or 5, characterized in that: The methods for carrying out the esterification reaction include: direct esterification with N,N'-dicyclohexylcarbodiimide and 4-dimethylaminopyridine, acetylation protection-acyl chloride-esterification-deacetylation, benzylation protection-acyl chloride or N,N'-dicyclohexylcarbodiimide esterification-hydrogenation debenzylation.

7. The preparation method according to claim 4, characterized in that: The molar ratio of the phenolic acid to the triterpene compound is 1:1-3; the organic solvent is one or more of dichloromethane, ethyl acetate or acetone; the solvent system of the silica gel column chromatography is any two or a combination of three of dichloromethane, ethyl acetate, acetone, methanol and water; the preparative liquid chromatography separation is separation using methanol and 0.5% formic acid aqueous solution at a volume ratio of 60:40, or separation using acetonitrile and 0.5% formic acid aqueous solution at a volume ratio of 70:

30.

8. The preparation method according to claim 5, characterized in that: The molar ratio of the phenolic acid to acetic anhydride is 1:1-3; the molar ratio of the acetylated phenolic acid to the triterpene compound is 1:3-5.

9. A pharmaceutical composition, characterized in that: The pharmaceutical composition contains a therapeutically effective amount of the triterpene phenolic acid ester derivative according to any one of claims 1 to 5.

10. Use of the triterpenoid phenolic acid ester derivative according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 9 in the preparation of a drug having the effect of regulating glucose and lipid metabolism.

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