CN111233810A - Preparation method and application of hydroxycinnamoyl ester type catechin - Google Patents

Preparation method and application of hydroxycinnamoyl ester type catechin Download PDF

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CN111233810A
CN111233810A CN202010056390.XA CN202010056390A CN111233810A CN 111233810 A CN111233810 A CN 111233810A CN 202010056390 A CN202010056390 A CN 202010056390A CN 111233810 A CN111233810 A CN 111233810A
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epigallocatechin
epicatechin
coumarate
butyldimethylsilyl
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鲍官虎
王威
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Anhui Agricultural University AHAU
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Abstract

The invention relates to a preparation method of hydroxycinnamoyl ester type catechin and application thereof, wherein the hydroxycinnamoyl ester type catechin comprises 4 kinds of catechin, namely epicatechin trans-coumarate, epicatechin trans-caffeate, epigallocatechin trans-coumarate and epigallocatechin trans-caffeate, the 4 kinds of hydroxycinnamoyl ester type catechin are prepared by respectively esterifying and removing protective groups of acetylated caffeic acid or acyl chloride of coumaric acid after the steps of total acetylation, acetyl removal on phenolic hydroxyl, phenolic hydroxyl silanization and 3-bit acetyl removal of epicatechin and epigallocatechin, the preparation method of the 4 kinds of catechin is simple and mild in condition, and can be completed under general experimental conditions, the 4 kinds of hydroxycinnamoyl ester type catechin has a certain inhibition effect on the activity of α -glucosidase, can be used for hypoglycemic drugs, and has important significance to the fields of agriculture and medicine.

Description

Preparation method and application of hydroxycinnamoyl ester type catechin
Technical Field
The invention belongs to the technical field of natural medicinal chemistry, and particularly relates to a preparation method and application of four hydroxycinnamoyl ester type catechins named as epicatechin trans-coumarate, epicatechin trans-caffeate, epigallocatechin trans-coumarate and epigallocatechin trans-caffeate respectively.
Background
The tea leaves contain rich tea polyphenol which accounts for 18 to 36 percent of dry substances of the tea leaves. Wherein most of catechin (flavanol) accounts for 12% -24% of dry matter of tea. Researches show that catechin is an important flavor substance in tea. In addition, catechin also has antiinflammatory, antioxidant, anticancer, cardiovascular and neuroprotective effects.
Chemical synthesis experiments usually require special reaction conditions, and the pharmaceutical reagents used in the reactions are generally dangerous. The synthesis method of the catechin disclosed in the prior art is complex and harsh in reaction conditions, and the conditions of a common laboratory cannot meet the reaction requirements and cannot complete the preparation. How to screen the existing preparation method and reaction raw materials, and the design of the preparation method of the four hydroxycinnamoyl ester type catechins with simple method and mild conditions has important significance.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects in the prior art and provide a preparation method of hydroxyl cinnamoyl ester catechin.
The invention also aims to solve the technical problem of providing the inhibition effect of the hydroxycinnamoyl ester type catechin prepared by the method on α -glucosidase, and the inhibition effect can be applied to the preparation of hypoglycemic drugs.
In order to solve the technical problem of the invention, the technical scheme is as follows:
a preparation method of hydroxycinnamoyl ester type catechin comprises 4 kinds of catechin, wherein the 4 kinds of catechin are epicatechin trans coumarate namely (-) -epicatechin 3-O-p-coumarate, epicatechin trans caffeate namely (-) -epicatechin 3-O-caffeate, epigallocatechin trans coumarate namely (-) -epigallocatechin 3-O-p-coumarate, epigallocatechin trans caffeate namely (-) -epigallocatechin 3-O-caffeate, the structural formulas of which are respectively shown as formula I, formula II, formula III and formula IV,
Figure BDA0002373026030000021
the preparation method of the hydroxycinnamoyl ester type catechin comprises the following steps:
s1, preparing 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin by using Epicatechin (EC) as a raw material through four reactions of total acetylation, acetyl removal on phenolic hydroxyl, phenolic hydroxyl silanization and 3-acetyl removal;
using Epigallocatechin (EGC) as a raw material, and preparing 5,7,3 ', 4 ', 5 ' -O-penta-tert-butyl dimethyl silicon-based-epigallocatechin through four reactions of full acetylation, acetyl removal on phenolic hydroxyl, phenolic hydroxyl silanization and 3-acetyl removal;
s2, preparing 3 ', 4' -O-diacetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate;
weighing 92.0mg of 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin obtained in step S1 in a 25mL round-bottomed flask, adding 0.2mL of dried pyridine and 2.0mL of dried dichloromethane, slowly adding dropwise 3.0mL of dried dichloromethane containing 0.18mmol of 3, 4-O-diacetyl caffeoyl chloride under ice bath conditions, reacting overnight at room temperature, adding an appropriate amount of water and an equal volume of dichloromethane for extraction three times, drying the dichloromethane layer with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography with petroleum ether: eluting with ethyl acetate ═ 30:1 to give 3 ", 4" -O-diacetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate;
s3, preparing 4 ' -O-acetyl-5, 7,3 ', 4 ' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-coumarate;
395.0mg of 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin obtained in step S1 was weighed into a 25mL round-bottomed flask, 1.5mL of dried pyridine and 3.0mL of dried dichloromethane were added, 5.0mL of dried dichloromethane containing 0.80mmol of 4-O-acetylated coumaroyl chloride was slowly added dropwise under ice bath conditions, the mixture was reacted overnight at room temperature, an appropriate amount of water and an equal volume of dichloromethane were added thereto to extract three times, the dichloromethane layer was dried over anhydrous sodium sulfate and then concentrated, and the resulting product was subjected to silica gel column chromatography with petroleum ether: eluting with ethyl acetate 50:1 to obtain 4 "-O-acetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-coumarate;
s4, preparing 3 ', 4 ' -O-diacetyl-5, 7,3 ', 4 ', 5 ' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-caffeate;
weighing 500.0mg of 5,7,3 ', 4 ', 5 ' -O-pentatert-butyldimethylsilyl-epigallocatechin obtained in step S1 in a 100mL round-bottomed flask, adding 2.0mL of dried pyridine and 5.0mL of dried dichloromethane, slowly adding 8.0mL of dried dichloromethane containing 1.9mmol of 3, 4-O-diacetyl caffeoyl chloride dropwise under ice bath conditions, reacting overnight at room temperature, adding an appropriate amount of water and an equal volume of dichloromethane for extraction three times, drying the dichloromethane layer with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography with petroleum ether: eluting with ethyl acetate (30: 1) to obtain 3 ', 4 ' -O-diacetyl-5, 7,3 ', 4 ', 5 ' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-caffeate;
s5, preparing 4 '-O-acetyl-5, 7, 3', 4 ', 5' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-coumarate;
weighing 370.0mg of 5,7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin obtained in step S1 in a 100mL round-bottom flask, adding 2.0mL of dried pyridine and 5.0mL of dried dichloromethane, slowly adding 7.0mL of dried dichloromethane containing 1.5mmol of 4-O-acetylated coumaroyl chloride dropwise under ice bath conditions, reacting overnight at room temperature, adding an appropriate amount of water and an equal volume of dichloromethane for extraction three times, drying the dichloromethane layer with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography with petroleum ether: eluting with ethyl acetate 50:1 to obtain 4 '-O-acetyl-5, 7, 3', 4 ', 5' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-coumarate;
s6, preparing epicatechin trans-caffeate;
weighing 164.0mg of 3 ' -O-diacetyl-5, 7,3 ', 4 ' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate prepared in the step S2 into a 25mL round-bottom flask, adding 28.0mg of potassium bifluoride and 6.0mL of dried methanol, reacting for 6 hours at 50 ℃, and performing LH-20 gel column chromatography on a reaction product to obtain epicatechin trans-caffeate after eluting with methanol;
s7, preparing epigallocatechin trans-caffeate;
weighing 60.0mg of 3 '-O-diacetyl-5, 7, 3', 4 ', 5' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-caffeate prepared in the step S4 into a 25mL round-bottom flask, adding 200.0mg of potassium bifluoride and 5.0mL of dried methanol, reacting at 50 ℃ for 6 hours, and performing LH-20 gel column chromatography on a reaction product to obtain epigallocatechin trans-caffeate after eluting with methanol;
s8, preparing epicatechin trans-coumarate;
163.0mg of 4 ' -O-acetyl-5, 7,3 ', 4 ' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-coumarate prepared in the step S3 was weighed into a 25mL round-bottomed flask, 136.0mg of potassium bifluoride was added, 6.0mL of dried methanol was added, reaction was performed at room temperature for 3.5 hours, an appropriate amount of water was added after the reaction was completed, extraction was performed three times with an equal volume of ethyl acetate, the ethyl acetate layer was dried over anhydrous sodium sulfate and then concentrated, and the mixture was subjected to silica gel column chromatography with petroleum ether: eluting with ethyl acetate 1:1 to obtain 4 "-O-acetyl-epicatechin trans-coumarate;
weighing 50.0mg of 4' -O-acetyl-epicatechin trans-coumarate in a 10mL round-bottom flask, adding 450.0mg of ammonium acetate, adding 2.0mL of 80% methanol aqueous solution by volume fraction, reacting at room temperature for 5.5 hours, performing LH-20 gel column chromatography on a reaction product, and eluting with methanol to obtain epicatechin trans-coumarate;
s9, preparing epigallocatechin trans-coumarate;
weighing 130.0mg of 4 ' -O-acetyl-5, 7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-coumarate prepared in the step S5 into a 25mL round-bottom flask, adding 476.0mg of potassium bifluoride, adding 8.0mL of dried methanol, reacting for 6 hours at 50 ℃, concentrating a reaction product, performing LH-20 gel column chromatography, and eluting with methanol to obtain 4 ' -O-acetyl-epigallocatechin trans-coumarate;
weighing 36.0mg of 4' -O-acetyl-epigallocatechin trans-coumarate in a 10mL round-bottom flask, adding 460.0mg of ammonium acetate, adding 2.0mL of 80% methanol aqueous solution by volume fraction, reacting at room temperature for 5 hours, carrying out LH-20 gel column chromatography on a reaction product, and eluting with methanol to obtain epigallocatechin trans-coumarate;
wherein, the steps S2, S3, S4 and S5 are not in sequence, and the steps S6, S7, S8 and S9 are not in sequence.
As a further improvement of the preparation method of the hydroxycinnamoyl ester type catechin,
preferably, the 3, 4-O-diacetyl caffeoyl chloride described in step S2 is prepared by:
180.1mg caffeic acid was weighed into a 25mL round bottom flask, 0.3mL dry pyridine, 0.5mL acetic anhydride was added and stirred overnight at room temperature under dry tube protection. After the reaction is finished, adding water to stop the reaction, and rotationally evaporating to dryness to obtain acetylated caffeic acid, namely 3, 4-O-diacetyl caffeic acid; transferring the acetylated caffeic acid after reaction to a 50mL round-bottom flask, adding 10mL thionyl chloride, heating and refluxing at 90 ℃ for 3 hours to obtain a crude product of 3, 4-O-diacetyl caffeoyl chloride, and storing for later use.
Preferably, the 4-O-acetylated coumaroyl chloride described in step S3 is prepared by:
656.0mg coumaric acid was weighed into a 25mL round bottom flask, 0.8mL dry pyridine, 1.0mL acetic anhydride was added, and stirred overnight at room temperature under dry tube protection. After the reaction is finished, adding water to stop the reaction, and obtaining acetylated coumaric acid after rotary evaporation to dryness; transferring the acetylated coumaric acid after reaction into a 25mL round-bottom flask, adding 3.0mL thionyl chloride, heating and refluxing at 90 ℃ for 3 hours to obtain a crude product of 4-O-acetylated coumaric acid chloride, and storing for later use.
In order to solve another technical problem, the invention provides another technical scheme that the hydroxycinnamoyl ester type catechin prepared by the preparation method is used for preparing the hypoglycemic medicament.
The application of the hydroxycinnamoyl ester type catechin in preparing the hypoglycemic medicament is further improved, the hypoglycemic medicament comprises an oral administration type and an injection type, the oral administration type comprises tablets, capsules, granules and dropping pills, and the injection type comprises injection and mixed rotary fluid.
Compared with the prior art, the invention has the beneficial effects that:
1) the invention provides a synthesis method of four hydroxycinnamoyl ester type catechins, which is different from the synthesis method in the prior art, the hydroxyl protecting groups, the esterification reaction conditions, the hydrolysis conditions of the protecting groups and the like adopted by the synthesis experiment have obvious advantages, the preparation method is simple, the conditions are mild, the implementation is easy, the cost is lower, and the invention has very good application prospect
2) The four hydroxycinnamoyl ester type catechins prepared by the invention have medical activity, have certain inhibition effect on α -glucosidase activity, can be used for preparing oral and injection type hypoglycemic drugs, and have important significance in the fields of agriculture and medicine.
Drawings
FIG. 1 shows the synthesis route of four hydroxycinnamoyl type catechins;
the designations in the drawings have the following meanings:
1a, 3,5,7,3 ', 4' -O-pentaacetyl-epicatechin
1b, 3,5,7,3 ', 4 ', 5 ' -O-hexaacetyl-epigallocatechin
2a, 3-O-acetyl-epicatechin
2b, 3-O-acetyl-epigallocatechin
3a, 3-O-acetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin
3b, 3-O-acetyl-5, 7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin
4a, 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin
4b, 5,7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin
5a, 3, 4-O-diacetyl caffeoyl chloride
5b, 4-O-acetylated coumaroyl chloride
6a, 3 ', 4' -O-diacetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate
6b, 4 "-O-acetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-coumarate
6c, 3 ', 4 ' -O-diacetyl-5, 7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-caffeate
6d, 4 '-O-acetyl-5, 7, 3', 4 ', 5' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-coumarate
7a, 4 "-O-acetyl-epicatechin trans-coumarate
7b, 4 "-O-acetyl-epigallocatechin trans-coumarate
8a, epicatechin trans-caffeate
8b epicatechin trans coumarate
8c Epigallocatechin Trans-caffeate
8d Epigallocatechin Trans coumarate
Detailed Description
The invention is further described below with reference to the following examples:
this section generally describes the materials used in the experiments of the present invention, as well as the experimental methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is described herein in detail as much as possible. It will be apparent to those skilled in the art that the materials, equipment, and methods of operation used in the present invention are well known in the art to which the invention pertains, unless otherwise specified.
Example 1
Preparing hydroxycinnamoyl catechin of epicatechin trans-coumarate, epicatechin trans-caffeate, epigallocatechin trans-coumarate and epigallocatechin trans-caffeate;
1.1 instruments and reagents
1The H nuclear magnetic resonance spectrum adopts an Agilent DD2600 MHz nuclear magnetic resonance instrument; the mass spectrum was obtained by Agilent6465 UPLC-Q-TOF-MS.
Epicatechin (EC) and Epigallocatechin (EGC) were purchased from Chengdu biopharmaceutical corporation and Hubei Jusheng technology corporation, respectively; caffeic acid and p-coumaric acid were purchased from annaiji chemistry; other reagents are all domestic analytical purifiers.
1.2 the synthetic route of hydroxycinnamic ester type catechin of epicatechin trans-coumarate, epicatechin trans-caffeate, epigallocatechin trans-coumarate and epicatechin trans-caffeate is shown in figure 1;
1.3 the specific implementation method comprises the following steps:
synthesis of compound 1 a:
1.16g (4.0mmol) of Epicatechin (EC) was weighed into a 50mL round bottom flask, 6.0mL of dried pyridine and 8.0mL of acetic anhydride (84.6mmol) were added, and the mixture was stirred at room temperature overnight under protection of a dry tube. After the reaction was completed, water was added to terminate the reaction, and the EC peracetylated product 3,5,7,3 ', 4' -O-pentaacetyl-epicatechin (1a,1.9g, white powder, yield 95%) was obtained after rotary evaporation to dryness.1H NMR(600MHz,CDCl3):7.33(1H,s),7.25(1H,d,J＝8.4Hz),7.18(1H,d,J＝8.4Hz),6.65(1H,br s),6.55(1H,br s),5.37(1H,br s),5.09(1H,s),2.95(1H,dd,J＝18.0,4.2Hz),2.87(1H,br d,J＝18.0Hz),2.27(9H,s),2.26(3H,s),1.90(3H,s)。
Synthesis of compound 1 b:
1.2g (3.9mmol) of Epigallocatechin (EGC) was weighed into a 50mL round-bottomed flask, 6.0mL of dried pyridine and 10.0mL of acetic anhydride (105.7mmol) were added, and the mixture was stirred at room temperature overnight under protection of a drying tube. After the reaction is finished, adding water to stop the reaction, and rotationally evaporating to dryness to obtain an EGC (epigallocatechin gallate) full acetylation product: 3,5,7,3 ', 4 ', 5 ' -O-hexaacetyl-epigallocatechin (1b, 2.1g, white powder, yield 94%).1H NMR(600MHz,CDCl3):7.20(2H,s),6.65(1H,brs),6.55(1H,br s),5.36(1H,br s),5.07(1H,s),2.94(1H,dd,J＝18.0,4.2Hz),2.88(1H,br d,J＝17.4Hz),2.25-2.28(15H,m),1.92(3H,s)。
Synthesis of compound 2 a:
500.0mg (1.0mmol) of Compound 1a was weighed into a 100mL round-bottomed flask, 50mL of 80% aqueous methanol solution and 7.8g (101.2mmol) of ammonium acetate were added, and the mixture was stirred at room temperatureStirring overnight. After the reaction is finished, adding a proper amount of water, removing methanol through rotary concentration, extracting for three times by using ethyl acetate with the same volume, concentrating an ethyl acetate layer, and performing silica gel column chromatography to obtain a product with ethyl acetate: the compound 3-O-acetyl-epicatechin (2a, 259.0mg, white powder, yield 78%) was obtained after elution with methanol ═ 20: 1.1HNMR(600MHz,DMSO-d6):9.29(1H,s),9.02(1H,s),8.92(1H,s),8.83(1H,s),6.83(1H,s),6.69(1H,d,J＝8.4Hz),6.65(1H,d,J＝7.8Hz),5.93(1H,s),5.76(1H,s),5.23(1H,br s),4.93(1H,s),2.87(1H,dd,J＝17.4,4.2Hz),2.58(1H,br d,J＝16.8Hz),1.87(3H,s)。
Synthesis of compound 2 b:
in a 500mL round-bottom flask, 4.2g (1.0mol) of Compound 1b was weighed, and 80mL of 80% aqueous methanol solution and 1.0g (11.9mmol) of sodium hydrogencarbonate were added and the mixture was stirred at room temperature overnight. After the reaction is finished, adding a proper amount of water, extracting for three times by using ethyl acetate with the same volume, concentrating an ethyl acetate layer, and performing silica gel column chromatography by using ethyl acetate: the compound 3-O-acetyl-epigallocatechin (2b, 1.05mg, white powder, yield 40%) was obtained after elution with methanol ═ 1: 1.1H NMR(600MHz,DMSO-d6):9.25(1H,s),8.99(1H,s),8.78(1H,s),7.99(1H,s),6.35(2H,s),5.92(1H,d,J＝1.2Hz),5.75(1H,d,J＝1.8Hz),5.21(1H,br s),4.85(1H,s),2.87(1H,dd,J＝17.4,4.8Hz),2.57(1H,br d,J＝16.8Hz),1.88(3H,s)。
Synthesis of compound 3 a:
200.0mg (0.6mmol) of Compound 2a are weighed into a 100mL round-bottomed flask, 10mL of dry dichloromethane, 816.0mg (12.0mmol) of imidazole, 40mg (0.33mmol) of 4-dimethylaminopyridine are added and 10mL of a dry dichloromethane solution containing 723.4mg (4.8mmol) of tert-butyldimethylchlorosilane is slowly added dropwise under ice-bath conditions. Stir at room temperature overnight. After the reaction is finished, adding a proper amount of water, extracting with dichloromethane with the same volume for three times, washing a dichloromethane layer solution with saturated sodium chloride, concentrating, and performing silica gel column chromatography with petroleum ether: the compound 3-O-acetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin (3a, 450.0mg, clear oil, yield 95%) was obtained after elution with ethyl acetate ═ 100: 1.1H NMR(600MHz,CDCl3):6.87(1H,s),6.85(1H,d,J＝8.4Hz),6.79(1H,d,J＝7.8Hz),6.13(1H,brs),5.96(1H,br s),5.37(1H,br s),4.94(1H,s),2.90(1H,dd,J＝17.4,4.2Hz),2.83(1H,br d,J＝18.0Hz),1.87(3H,s),0.94-0.98(36H,m),0.15-0.21(24H,m)。
Synthesis of compound 3 b:
500.0mg (1.4mmol) of Compound 2b is weighed into a 100mL round-bottom flask, 10mL of dry dichloromethane, 2.4g (35.3mmol) of imidazole, 100mg (0.82mmol) of 4-dimethylaminopyridine are added and 10mL of a solution of dry dichloromethane containing 2.4g (16.0mmol) of tert-butyldimethylchlorosilane is slowly added dropwise under ice-bath conditions. Stir at room temperature overnight. After the reaction is finished, adding a proper amount of water, extracting with dichloromethane with the same volume for three times, washing a dichloromethane layer solution with saturated sodium chloride, concentrating, and performing silica gel column chromatography with petroleum ether: the compound 3-O-acetyl-5, 7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin (3b, 970.0mg, clear oil, yield 74%) was obtained after elution from ethyl acetate ═ 200: 1.1HNMR(600MHz,CDCl3):6.55(2H,s),6.11(1H,d,J＝2.4Hz),5.95(1H,d,J＝2.4Hz),5.36(1H,m),4.91(1H,br s),2.89(1H,dd,J＝17.4,4.8Hz),2.81(1H,dd,J＝17.4,3.0Hz),1.86(3H,s),0.90-0.97(45H,m),0.08-0.21(30H,m)。
Synthesis of compound 4 a:
100.0mg (0.13mmol) of Compound 3a was weighed into a 50mL round-bottomed flask, dissolved in 100uL of dry dichloromethane, and then 10mL of dry methanol and 50.0mg (0.36mmol) of potassium carbonate were added, followed by stirring at room temperature for 2 hours. After the reaction is finished, adding a proper amount of water, extracting for three times by using ethyl acetate with the same volume, drying and concentrating ethyl acetate layer solution by using anhydrous sodium sulfate, and performing silica gel column chromatography by using petroleum ether: the compound 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin (4a, 38.0mg, clear oil, yield 40%) was obtained after elution from ethyl acetate ═ 100: 1.1H NMR(600MHz,CDCl3):6.94(1H,d,J＝1.8Hz),6.92(1H,dd,J＝8.4,1.8Hz),6.84(1H,d,J＝7.8Hz),6.11(1H,d,J＝2.4Hz),5.96(1H,d,J＝1.8Hz),4.86(1H,s),4.20(1H,br s),2.86(1H,s),2.85(1H,s),0.99(9H,s),0.97(9H,s),0.96(9H,s),0.95(9H,s),0.16-0.22(24H,m)。
Synthesis of compound 4 b:
970.0mg (1.06mmol) of compound 3b was weighed into a 100mL round-bottom flask, dissolved in 100uL of dry dichloromethane, and then 40mL of dry methanol and 200mg (1.44mmol) of potassium carbonate were added, followed by stirring at room temperature for 2 hours. After the reaction is finished, adding a proper amount of water, extracting for three times by using ethyl acetate with the same volume, drying and concentrating ethyl acetate layer solution by using anhydrous sodium sulfate, and performing silica gel column chromatography by using petroleum ether: the compound 5,7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin (4b, 353.0mg, clear oil, yield 38%) was obtained by eluting with ethyl acetate (200: 1).1H NMR(600MHz,CDCl3):6.59(2H,s),6.09(1H,d,J＝1.8Hz),5.95(1H,d,J＝2.4Hz),4.82(1H,br s),4.16(1H,m),3.47(1H,s),2.85(1H,dd,J＝16.8,4.2Hz),2.80(1H,dd,J＝16.8,3.0Hz),0.90-0.99(45H,m),0.10-0.22(30H,m)。
Synthesis of compound 5 a:
180.1mg (1.0mmol) of caffeic acid was weighed into a 25mL round bottom flask, 0.3mL of dried pyridine, 0.5mL of acetic anhydride (5.3mmol) were added, and the mixture was stirred at room temperature overnight under the protection of a drying tube. After the reaction was completed, water was added to terminate the reaction, and the reaction was rotary-evaporated to dryness to obtain acetylated caffeic acid (3, 4-O-diacetyl caffeic acid, 250.8mg, yield 95%).1H NMR(600MHz,DMSO-d6) 12.47(1H, s)7.66(1H, s),7.63(1H, d, J ═ 8.4Hz),7.57(1H, d, J ═ 16.2Hz),7.31(1H, d, J ═ 8.4Hz),6.53(1H, d, J ═ 16.2Hz),2.29(3H, s). Transferring the acetylated caffeic acid after reaction to a 50mL round-bottom flask, adding 10mL thionyl chloride, heating and refluxing at 90 ℃ for 3 hours to obtain a crude compound 3, 4-O-diacetyl caffeoyl chloride (5a), and directly using the crude compound in the next reaction.
Synthesis of compound 5 b:
656.0mg (4.0mmol) of caffeic acid was weighed into a 25mL round bottom flask, 0.8mL of dried pyridine, 1.0mL of acetic anhydride (10.6mmol) were added, and the mixture was stirred at room temperature overnight under protection of a drying tube. After the reaction was completed, water was added to terminate the reaction, and the reaction was rotary-evaporated to dryness to obtain acetylated coumaric acid (4-O-acetylated coumaric acid, 774.6mg, yield 94%).1H NMR(600MHz,DMSO-d6):12.36(1H,s)7.73(2H, d, J ═ 9.0Hz),7.60(1H, d, J ═ 14.4Hz),7.18(2H, d, J ═ 8.4Hz),6.50(1H, d, J ═ 16.2Hz),2.28(3H, s). The acetylated coumaric acid (251.0mg, 1.2mmol) after the reaction was transferred to a 25mL round bottom flask, 3.0mL thionyl chloride was added, and the mixture was heated under reflux at 90 ℃ for 3 hours to obtain the crude compound 4-O-acetylated coumaroyl chloride (5b), which was used directly in the next reaction.
Synthesis of compound 6 a:
compound 4a (92.0mg, 0.12mmol) was weighed into a 25mL round bottom flask, 0.2mL dry pyridine, 2.0mL dry dichloromethane were added, and 3.0mL dry dichloromethane containing compound 5a (0.18mmol) was slowly added dropwise under ice bath conditions. Reacting at room temperature overnight, adding a proper amount of water and dichloromethane with the same volume, extracting for three times, drying a dichloromethane layer by using anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography by using petroleum ether: the compound 3 ", 4" -O-diacetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate (6a, 95.5mg, oil, 78% yield) was obtained after elution with ethyl acetate ═ 30: 1.1H NMR(600MHz,CDCl3):7.45(1H,d,J＝15.6Hz),7.32(1H,dd,J＝8.4,1.2Hz),7.26(1H,br s),7.16(1H,d,J＝8.4Hz),6.89(1H,d,J＝1.8Hz),6.87(1H,br d,J＝8.4Hz),6.77(1H,d,J＝8.4Hz),6.26(1H,d,J＝16.2Hz),6.15(1H,d,J＝2.4Hz),5.96(1H,d,J＝1.8Hz),5.50(1H,br s),5.00(1H,s),2.96(1H,dd,J＝18,4.8Hz),2.90(1H,br d,J＝17.4Hz),2.27(6H,s),0.92-0.97(36H,m),0.12-0.21(24H,m)。
Synthesis of compound 6 b:
compound 4a (395.0mg, 0.53mmol) was weighed into a 25mL round bottom flask, 1.5mL of dried pyridine, 3.0mL of dried dichloromethane were added, and 5.0mL of dried dichloromethane containing compound 5b (0.80mmol) was slowly added dropwise under ice bath conditions. Reacting at room temperature overnight, adding appropriate amount of water and dichloromethane with equal volume, extracting for three times, drying dichloromethane layer with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography with petroleum ether: the compound 4 "-O-acetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans coumarate (6b, 197.8mg, oil, yield 40%) was obtained after elution with ethyl acetate ═ 50: 1.1H NMR(600MHz,CDCl3):7.49(1H,d,J＝15.6Hz),7.44(2H,d,J＝9.0Hz),7.07(1H,d,J＝9.0Hz),6.91(1H,d,J＝1.8Hz),6.86(1H,dd,J＝8.4,2.4Hz),6.77(1H,d,J＝8.4Hz),6.26(1H,d,J＝16.2Hz),6.16(1H,d,J＝1.8Hz),5.97(1H,d,J＝2.4Hz),5.52(1H,br s),5.01(1H,s),2.97(1H,dd,J＝17.4,4.2Hz),2.90(1H,dd,J＝16.8,3.0Hz),2.28(3H,s),0.92-0.97(36H,m),0.12-0.21(24H,m)。
Synthesis of compound 6 c:
compound 4b (500.0mg, 0.57mmol) was weighed into a 100mL round-bottomed flask, 2.0mL of dried pyridine and 5.0mL of dried dichloromethane were added, and 8.0mL of dried dichloromethane containing compound 5a (1.9mmol) was slowly added dropwise under ice-bath conditions. Reacting at room temperature overnight, adding a proper amount of water and dichloromethane with the same volume, extracting for three times, drying a dichloromethane layer by using anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography by using petroleum ether: the compound 3 ", 4" -O-diacetyl-5, 7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-caffeate (6c, 172.1mg, oil, yield 70%) was obtained after elution with ethyl acetate ═ 30: 1.1H NMR(600MHz,CDCl3):7.44(1H,d,J＝16.2Hz),7.29(1H,dd,J＝8.4,1.8Hz),7.24(1H,s),7.16(1H,d,J＝8.4Hz),6.56(2H,s),6.23(1H,d,J＝16.2Hz),6.14(1H,d,J＝2.4Hz),5.95(1H,d,J＝2.4Hz),5.49(1H,m),4.97(1H,s),2.95(1H,dd,J＝17.4,4.8Hz),2.87(1H,dd,J＝17.4,3.6Hz),2.26(6H,s),0.86-0.97(45H,m),0.04-0.21(30H,m)。
Synthesis of compound 6 d:
compound 4b (370.0mg, 0.42mmol) was weighed into a 100mL round bottom flask, 2.0mL dry pyridine and 5.0mL dry dichloromethane were added, and 7.0mL dry dichloromethane containing compound 5b (1.5mmol) was slowly added dropwise under ice bath conditions. Reacting at room temperature overnight, adding a proper amount of water and dichloromethane with the same volume, extracting for three times, drying a dichloromethane layer by using anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography by using petroleum ether: the compound 4 "-O-acetyl-5, 7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-coumarate (6d, 280.0mg, oil, yield 62%) was obtained after elution with ethyl acetate ═ 50: 1.1H NMR(600MHz,CDCl3):7.48(1H,d,J＝16.2Hz),7.43(2H,d,J＝9.0Hz),7.07(2H,d,J＝8.4Hz),6.57(2H,s),6.24(1H,d,J＝15.6Hz),6.15(1H,d,J＝1.8Hz),5.96(1H,d,J＝2.4Hz),5.50(1H,m),4.97(1H,s),2.95(1H,dd,J＝17.4,4.8Hz),2.88(1H,dd,J＝17.4,3.0Hz),2.28(3H,s),0.86-0.97(45H,m),0.04-0.20(30H,m)。
Compound 7a synthesis:
compound 6b (163.0mg, 0.17mmol) was weighed into a 25mL round-bottomed flask, potassium bifluoride (136.0mg, 1.7mmol) was added, 6.0mL of dried methanol was further added, and the reaction was carried out at room temperature for 3.5 hours. After the reaction is finished, adding a proper amount of water, and extracting for three times by using ethyl acetate with the same volume. The ethyl acetate layer was dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography with petroleum ether: the compound 4 "-O-acetyl-epicatechin trans-coumarate (7a, 70.9mg, white powder, yield 85%) was obtained after elution with ethyl acetate ═ 1: 1.1H NMR(600MHz,DMSO-d6):9.27(1H,s),9.01(1H,s),8.82(1H,s),8.77(1H,s),7.73(2H,d,J＝8.4Hz),7.51(1H,dd,J＝16.2Hz),7.15(2H,d,J＝8.4Hz),6.89(1H,d,J＝1.8Hz),6.71(1H,dd,J＝8.4,1.8Hz),6.67(1H,d,J＝8.4Hz),6.48(1H,d,J＝16.2Hz),5.94(1H,d,J＝1.8Hz),5.81(1H,d,J＝1.8Hz),5.36(1H,br s),5.02(1H,s),2.95(1H,dd,J＝17.4,4.8Hz),2.68(1H,br d,J＝16.8Hz),2.27(3H,s)。
Compound 7b synthesis:
compound 6d (130.0mg, 0.12mmol) was weighed into a 25mL round-bottomed flask, potassium bifluoride (476.0mg, 6.1mmol) was added, 8.0mL of dried methanol was further added, and the reaction was carried out at 50 ℃ for 6 hours. The reaction product was concentrated and subjected to LH-20 gel column chromatography to give 4 "-O-acetyl-epigallocatechin trans-coumarate (7b, 36.0mg, white powder, yield 60%) as a compound after elution with methanol.1H NMR(600MHz,DMSO-d6):9.29(1H,s),9.03(1H,s),8.76(1H,s),8.00(1H,s),7.72(2H,d,J＝8.4Hz),7.51(1H,dd,J＝16.2Hz),7.15(2H,d,J＝9.0Hz),6.46(1H,d,J＝16.2Hz),6.40(2H,s),5.94(1H,d,J＝2.4Hz),5.79(1H,d,J＝2.4Hz),5.34(1H,brs),4.95(1H,s),2.94(1H,dd,J＝17.4,4.8Hz),2.67(1H,br d,J＝16.2Hz),2.26(3H,s)。
Compound 8a synthesis:
6a (164.0mg, 0.16mmol) was weighed into a 25mL round bottom flask and addedPotassium bifluoride (28.0mg, 1.64mmol) was added to 6.0mL of dry methanol, and the mixture was reacted at 50 ℃ for 6 hours. The reaction product was subjected to LH-20 gel column chromatography and eluted with methanol to give the compound epicatechin trans-caffeate (8a, 52.0mg, white powder, yield 70%).1H NMR(600MHz,DMSO-d6) 7.32(1H, d, J ═ 16.2Hz),6.98(1H, s),6.95(1H, dd, J ═ 6.6,1.8Hz),6.88(1H, s),6.73(1H, d, J ═ 7.8Hz),6.70(1H, dd, J ═ 8.4,1.2Hz),6.67(1H, dd, J ═ 7.8Hz),6.12(1H, d, J ═ 15.6Hz),5.94(1H, d, J ═ 1.8Hz),5.81(1H, d, J ═ 1.8Hz),5.31(1H, br), 5.01(1H, s),2.92(1H, dd, J ═ 17.4,4.8Hz), 2.92(1H, dd, J ═ 2.67, 16H, br). HR-ESI-MS (negative ion) M/z451.1040[ M-H]-。
Synthesis of compound 8 b:
in a 10mL round-bottom flask, 7a (50.0mg, 0.1mmol) was weighed, ammonium acetate (450.0mg, 5.8mmol) was added, and 2.0mL of an 80% aqueous methanol solution was added and reacted at room temperature for 5.5 hours. The reaction product was subjected to LH-20 gel column chromatography and eluted with methanol to give the compound epicatechin trans coumarate (8b, 45.6mg, white powder, yield 88%). 1H NMR (600MHz, DMSO-d)6) 7.50(2H, d, J ═ 8.4Hz),7.41(1H, d, J ═ 15.6Hz),6.88(1H, d, J ═ 1.8Hz),6.75(2H, d, J ═ 9.0Hz),6.70(1H, dd, J ═ 8.4,1.2Hz),6.67(1H, d, J ═ 7.8Hz),6.24(1H, d, J ═ 15.6Hz),5.93(1H, d, J ═ 1.8Hz),5.80(1H, d, J ═ 1.8Hz),5.32(1H, br), 5.01(1H, s),2.92(1H, dd, J ═ 17.4,4.8Hz),2.66(1H, br, 16.16 Hz). HR-ESI-MS (negative ion) M/z435.1088[ M-H]-。
Synthesis of compound 8 c:
6c (60.0mg, 0.05mmol) was weighed into a 25mL round-bottomed flask, potassium bifluoride (200.0mg, 2.56mmol) was added, 5.0mL of dried methanol was further added, and the reaction was carried out at 50 ℃ for 6 hours. The reaction product was subjected to LH-20 gel column chromatography and eluted with methanol to give the compound epigallocatechin trans-caffeate (8c, 15.0mg, white powder, yield 60%).1H NMR(600MHz,DMSO-d6):7.33(1H,d,J＝16.8Hz),6.98(1H,d,J＝1.8Hz),6.96(1H,dd,J＝8.4,1.8Hz),6.72(1H,d,J＝8.4Hz),6.39(2H,s),6.09(1H,d,J＝16.2Hz),5.93(1H,d,J＝1.8Hz),5.79(1H,d,J＝2.4Hz),5.28(1H,br s),4.93(1H,s),2.90(1H,dd,J＝17.4,4.8Hz),2.64(1H,br d,J＝16.2Hz)。HR-ESI-MS (negative ion) M/z 451.1033[ M-H]-。
Synthesis of compound 8 d:
in a 10mL round-bottom flask, 7b (36.0mg, 0.07mmol) was weighed, ammonium acetate (460.0mg, 6.0mmol) was added, and 2.0mL of an 80% aqueous methanol solution was added and reacted at room temperature for 5 hours. The reaction product was subjected to LH-20 gel column chromatography and eluted with methanol to give the compound epigallocatechin trans-coumarate (8d, 18.1mg, white powder, yield 55%).1H NMR(600MHz,DMSO-d6) 7.49(2H, d, J ═ 8.4Hz),7.41(1H, d, J ═ 15.6Hz),6.75(2H, d, J ═ 8.4Hz),6.39(2H, s),6.22(1H, d, J ═ 15.6Hz),5.93(1H, d, J ═ 2.4Hz),5.79(1H, d, J ═ 1.8Hz),5.30(1H, br), 4.93(1H, s),2.91(1H, dd, J ═ 17.4,4.8Hz),2.65(1H, br, J ═ 16.2 Hz). HR-ESI-MS (negative ion) M/z467.0979[ M-H]-。
Example 2
The four hydroxycinnamoyl-type catechins, epicatechin trans-coumarate, epigallocatechin trans-coumarate and epigallocatechin trans-caffeate, prepared in example 1 were subjected to α -glucosidase activity inhibition experiments in vitro.
2.1 Experimental materials and reagents
Acarbose, potassium phosphate buffer (PPBS, 10mM, pH 6.9), α -glucosidase (0.1U. mL)-1) 4-Nitrophenyl- α -D-glucopyranoside (pNPG, 2.5mM), dimethyl sulfoxide (DMSO)
2.2 Experimental methods and results
Adding α -glucosidase solution of 60 μ L into a 96-well plate, adding four hydroxycinnamoyl ester type catechins of 60 μ L with different concentration gradients respectively, after pre-incubation for 10min at 37 ℃, adding pNPG solution of 60 μ L respectively, after further incubation for 20min at 37 ℃, placing the mixture in an enzyme-linked immunosorbent assay (25 ℃), recording the A value of the reaction solution under visible light with the wavelength of 405nm, repeating all the samples for 3 times, marking as A sample, replacing α -glucosidase with PPBS of 60 μ L, marking as A blank (repeating for 3 times), performing blank zero adjustment, replacing the sample solution with PPBS of 60 μ L, marking as A test, replacing the sample solution and α -glucosidase with PPBS of 120 μ L, marking as A control (repeating for 3 times).
The enzyme inhibition ratio of acetylcholinesterase was calculated according to the following formula:
the inhibition ratio (%) × (1- (sample a-a blank)/(test a-a control)) × 100%.
Plotting the relative activity of enzyme against the concentration of the inhibitor, and calculating the IC of the action of the four hydroxycinnamoyl ester catechins and α -glucosidase according to the inhibition curve50The value is obtained. The results obtained are shown in the following table:
Figure BDA0002373026030000161
note: positive control drug acarbose
The results in the table show that the four hydroxycinnamic acid ester type catechins prepared by the invention, namely epicatechin trans-coumarate, epicatechin trans-caffeate, epigallocatechin trans-coumarate and epigallocatechin trans-caffeate, have strong inhibition effect on α -glucosidase, can be applied to preparation of hypoglycemic drugs and have wide application prospect.
It should be understood that the examples and embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the present disclosure, and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this disclosure.

Claims (5)

1. A preparation method of hydroxycinnamoyl ester type catechin comprises 4 kinds of catechin, wherein the 4 kinds of catechin are epicatechin trans coumarate namely (-) -epicatechin 3-O-p-coumarate, epicatechin trans caffeate namely (-) -epicatechin 3-O-caffeate, epigallocatechin trans coumarate namely (-) -epigallocatechin 3-O-p-coumarate, epigallocatechin trans caffeate namely (-) -epigallocatechin 3-O-p-caffeate, and the structural formulas of the epigallocatechin trans caffeate are respectively shown as formula I, formula II, formula III and formula IV,
Figure FDA0002373026020000011
the preparation method of the hydroxycinnamoyl ester type catechin is characterized by comprising the following steps:
s1, preparing 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin by using Epicatechin (EC) as a raw material through four reactions of total acetylation, acetyl removal on phenolic hydroxyl, phenolic hydroxyl silanization and 3-acetyl removal;
using Epigallocatechin (EGC) as a raw material, and preparing 5,7,3 ', 4 ', 5 ' -O-penta-tert-butyl dimethyl silicon-based-epigallocatechin through four reactions of full acetylation, acetyl removal on phenolic hydroxyl, phenolic hydroxyl silanization and 3-acetyl removal;
s2, preparing 3 ', 4' -O-diacetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate;
weighing 92.0mg of 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin obtained in step S1 in a 25mL round-bottomed flask, adding 0.2mL of dried pyridine and 2.0mL of dried dichloromethane, slowly adding dropwise 3.0mL of dried dichloromethane containing 0.18mmol of 3, 4-O-diacetyl caffeoyl chloride under ice bath conditions, reacting overnight at room temperature, adding an appropriate amount of water and an equal volume of dichloromethane for extraction three times, drying the dichloromethane layer with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography with petroleum ether: eluting with ethyl acetate ═ 30:1 to give 3 ", 4" -O-diacetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate;
s3, preparing 4 ' -O-acetyl-5, 7,3 ', 4 ' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-coumarate;
395.0mg of 5,7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin obtained in step S1 was weighed into a 25mL round-bottomed flask, 1.5mL of dried pyridine and 3.0mL of dried dichloromethane were added, 5.0mL of dried dichloromethane containing 0.80mmol of 4-O-acetylated coumaroyl chloride was slowly added dropwise under ice bath conditions, the mixture was reacted overnight at room temperature, an appropriate amount of water and an equal volume of dichloromethane were added thereto to extract three times, the dichloromethane layer was dried over anhydrous sodium sulfate and then concentrated, and the resulting product was subjected to silica gel column chromatography with petroleum ether: eluting with ethyl acetate 50:1 to obtain 4 "-O-acetyl-5, 7,3 ', 4' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-coumarate;
s4, preparing 3 ', 4 ' -O-diacetyl-5, 7,3 ', 4 ', 5 ' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-caffeate;
weighing 500.0mg of 5,7,3 ', 4 ', 5 ' -O-pentatert-butyldimethylsilyl-epigallocatechin obtained in step S1 in a 100mL round-bottomed flask, adding 2.0mL of dried pyridine and 5.0mL of dried dichloromethane, slowly adding 8.0mL of dried dichloromethane containing 1.9mmol of 3, 4-O-diacetyl caffeoyl chloride dropwise under ice bath conditions, reacting overnight at room temperature, adding an appropriate amount of water and an equal volume of dichloromethane for extraction three times, drying the dichloromethane layer with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography with petroleum ether: eluting with ethyl acetate (30: 1) to obtain 3 ', 4 ' -O-diacetyl-5, 7,3 ', 4 ', 5 ' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-caffeate;
s5, preparing 4 '-O-acetyl-5, 7, 3', 4 ', 5' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-coumarate;
weighing 370.0mg of 5,7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin obtained in step S1 in a 100mL round-bottom flask, adding 2.0mL of dried pyridine and 5.0mL of dried dichloromethane, slowly adding 7.0mL of dried dichloromethane containing 1.5mmol of 4-O-acetylated coumaroyl chloride dropwise under ice bath conditions, reacting overnight at room temperature, adding an appropriate amount of water and an equal volume of dichloromethane for extraction three times, drying the dichloromethane layer with anhydrous sodium sulfate, concentrating, and performing silica gel column chromatography with petroleum ether: eluting with ethyl acetate 50:1 to obtain 4 '-O-acetyl-5, 7, 3', 4 ', 5' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-coumarate;
s6, preparing epicatechin trans-caffeate;
weighing 164.0mg of 3 ' -O-diacetyl-5, 7,3 ', 4 ' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-caffeate prepared in the step S2 into a 25mL round-bottom flask, adding 28.0mg of potassium bifluoride and 6.0mL of dried methanol, reacting for 6 hours at 50 ℃, and performing LH-20 gel column chromatography on a reaction product to obtain epicatechin trans-caffeate after eluting with methanol;
s7, preparing epigallocatechin trans-caffeate;
weighing 60.0mg of 3 '-O-diacetyl-5, 7, 3', 4 ', 5' -O-pentatert-butyldimethylsilyl-epigallocatechin trans-caffeate prepared in the step S4 into a 25mL round-bottom flask, adding 200.0mg of potassium bifluoride and 5.0mL of dried methanol, reacting at 50 ℃ for 6 hours, and performing LH-20 gel column chromatography on a reaction product to obtain epigallocatechin trans-caffeate after eluting with methanol;
s8, preparing epicatechin trans-coumarate;
163.0mg of 4 ' -O-acetyl-5, 7,3 ', 4 ' -O-tetra-tert-butyldimethylsilyl-epicatechin trans-coumarate prepared in the step S3 was weighed into a 25mL round-bottomed flask, 136.0mg of potassium bifluoride was added, 6.0mL of dried methanol was added, reaction was performed at room temperature for 3.5 hours, an appropriate amount of water was added after the reaction was completed, extraction was performed three times with an equal volume of ethyl acetate, the ethyl acetate layer was dried over anhydrous sodium sulfate and then concentrated, and the mixture was subjected to silica gel column chromatography with petroleum ether: eluting with ethyl acetate 1:1 to obtain 4 "-O-acetyl-epicatechin trans-coumarate;
weighing 50.0mg of 4' -O-acetyl-epicatechin trans-coumarate in a 10mL round-bottom flask, adding 450.0mg of ammonium acetate, adding 2.0mL of 80% methanol aqueous solution by volume fraction, reacting at room temperature for 5.5 hours, performing LH-20 gel column chromatography on a reaction product, and eluting with methanol to obtain epicatechin trans-coumarate;
s9, preparing epigallocatechin trans-coumarate;
weighing 130.0mg of 4 ' -O-acetyl-5, 7,3 ', 4 ', 5 ' -O-penta-tert-butyldimethylsilyl-epigallocatechin trans-coumarate prepared in the step S5 into a 25mL round-bottom flask, adding 476.0mg of potassium bifluoride, adding 8.0mL of dried methanol, reacting for 6 hours at 50 ℃, concentrating a reaction product, performing LH-20 gel column chromatography, and eluting with methanol to obtain 4 ' -O-acetyl-epigallocatechin trans-coumarate;
weighing 36.0mg of 4' -O-acetyl-epigallocatechin trans-coumarate in a 10mL round-bottom flask, adding 460.0mg of ammonium acetate, adding 2.0mL of 80% methanol aqueous solution by volume fraction, reacting at room temperature for 5 hours, carrying out LH-20 gel column chromatography on a reaction product, and eluting with methanol to obtain epigallocatechin trans-coumarate;
wherein, the steps S2, S3, S4 and S5 are not in sequence, and the steps S6, S7, S8 and S9 are not in sequence.
2. The method for preparing hydroxycinnamoyl ester type catechins of claim 1, wherein the 3, 4-O-diacetyl caffeoyl chloride of step S2 is prepared by:
weighing 180.1mg of caffeic acid into a 25mL round-bottom flask, adding 0.3mL of dried pyridine and 0.5mL of acetic anhydride, stirring at room temperature under the protection of a drying tube overnight, adding water to stop the reaction, and rotationally evaporating to dryness to obtain acetylated caffeic acid, namely 3, 4-O-diacetyl caffeic acid; transferring the acetylated caffeic acid after reaction to a 50mL round-bottom flask, adding 10mL thionyl chloride, heating and refluxing at 90 ℃ for 3 hours to obtain 3, 4-O-diacetyl caffeoyl chloride, and storing for later use.
3. The method for preparing hydroxycinnamoyl ester type catechins according to claim 1, wherein the 4-O-acetylated coumaroyl chloride described in step S3 is prepared by:
weighing 656.0mg coumaric acid in a 25mL round-bottom flask, adding 0.8mL dry pyridine and 1.0mL acetic anhydride, stirring at room temperature under the protection of a drying tube overnight, adding water to stop the reaction, and performing rotary evaporation to obtain acetylated coumaric acid; transferring the acetylated coumaric acid after reaction into a 25mL round-bottom flask, adding 3.0mL thionyl chloride, heating and refluxing at 90 ℃ for 3 hours to obtain 4-O-acetylated coumaric acid chloride, and storing for later use.
4. The application of the hydroxycinnamoyl ester type catechin prepared by the preparation method of claim 1 in preparing hypoglycemic drugs.
5. The use of the hydroxycinnamoyl ester type catechin prepared by the preparation method of claim 1 in preparing the hypoglycemic medicament according to claim 4, wherein the hypoglycemic medicament comprises an oral type and an injection type, the oral type comprises tablets, capsules, granules and dripping pills, and the injection type comprises injections and racemic mixtures.
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