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Preparation method of dammarane saponin and application of dammarane saponin in preparation of hypoglycemic and anti-inflammatory drugs and health products

A dammarane type, anti-inflammatory drug technology, applied in the field of biomedicine, to achieve the effect of good hypoglycemic and anti-inflammatory activities

Active Publication Date: 2020-08-25
GUANGXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the hypoglycemic and anti-inflammatory activities of monomeric compounds in Cyclocarya paliurus

Method used

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  • Preparation method of dammarane saponin and application of dammarane saponin in preparation of hypoglycemic and anti-inflammatory drugs and health products
  • Preparation method of dammarane saponin and application of dammarane saponin in preparation of hypoglycemic and anti-inflammatory drugs and health products
  • Preparation method of dammarane saponin and application of dammarane saponin in preparation of hypoglycemic and anti-inflammatory drugs and health products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] A method for preparing dammarane-type saponins, comprising the following steps:

[0100] S1: Take 10 kg of dried Cyclocarya paliurus leaves, add 75% ethanol aqueous solution, heat and reflux for extraction 3 times in sequence for 3 hours, 2 hours, and 2 hours, combine the filtrates, recover ethanol under reduced pressure, and obtain 2.10 kg of extract;

[0101] S2: Suspend 1.5kg of extract in 20L of water, absorb on D101 macroporous adsorption resin chromatographic column, wash with water, and then elute with 30% and 95% ethanol to obtain 250g and 625.0 g of eluate of 30% and 95% ethanol respectively g, The eluate is taken as the Cyclocarya paliurus leaf extract rich in dammarane-type triterpene saponins.

Embodiment 2

[0103] Isolation, preparation and structure identification of dammarane-type triterpene saponins 1-11.

[0104] Compounds 1-11 were isolated simultaneously and thus described in one example.

[0105] [Isolation and preparation of compound 1-11]

[0106] Get 95% ethanol eluate 150.0g obtained in the above-mentioned Example 1, through silica gel column chromatography, dichloromethane-methanol (100:0, 60:1, 20:1, 10:1, 8:1 , 4:1, 2:1, 0:100) as the eluent for gradient elution to obtain Fr.A (22g), Fr.B (16g), Fr.C (23g), Fr.D (15g) , Fr.E (27g), Fr.F (19g), Fr.G (13g) and Fr.H (6g). Component Fr.F (19g) was subjected to reverse-phase C18 silica gel column chromatography, using water-methanol (100:0-0:100) as the eluent for gradient elution, and 6 components were obtained as Fr.F. 1(0.5g), Fr.F.2(3.5g), Fr.F.3(4.2g), Fr.F.4(2.5g), Fr.F.5(1.5g) and Fr.F. 6 (1.1 g). Component Fr.F.3 (4.2g) was subjected to reverse phase C18 silica gel column chromatography (MeOH-H2O, 60:40→100:...

Embodiment 3

[0133]Determination of biological activity of dammarane-type triterpene saponins 1-11

[0134] 3.1 Compound 1-11 inhibits α-glucosidase activity and the determination of compound 1 and 2 on enzyme activity and reaction time

[0135] 3.1.1 Determination of inhibition rate of α-glucosidase activity

[0136] In the test, acarbose was used as the positive control drug, and all the compounds to be tested were diluted with phosphate buffer (pH=6.9) to different concentrations (0.001, 0.01, 0.05, 0.1, 0.2, 0.5 mg·mL －1 ) sample solution. In a 96-well plate, add 50 μL of compound solutions of various concentrations, 50 μL of phosphate buffer, and then add 100 μL of 0.2 U·mL －1 α-glucosidase solution, incubated at 37°C for 10min, then added 50μL 2mmol·L －1 The p-nitrophenyl-β-D-galactopyranoside (PNPG) was further incubated at 37°C for 5 minutes, and finally the absorbance (A) value was measured at a wavelength of 405nm. Inhibition rate (%) = [A 1 -(A 2 -A 3 )] / A 1 *100%, where...

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Abstract

The invention discloses a preparation method of dammarane saponin and application of the dammarane saponin in preparation of hypoglycemic and anti-inflammatory drugs and health products. The dammaranesaponin is prepared by the following operation steps of: S1, taking 10kg of dried cyclocarya paliurus leaves, adding a 75% ethanol aqueous solution, sequentially heating, refluxing and extracting for3 times for 3 hours, 2 hours and 2 hours respectively, combining filtrates, and recovering ethanol under reduced pressure to obtain 2.10kg of extract; and S2, taking and suspending 1.5kg of the extract in 20L of water, performing adsorption by using a D101 macroporous adsorption resin chromatographic column, performing washing by using water, and performing eluting by using 30% and 95% ethanol torespectively obtain 250g of 30% ethanol eluate and 625.0g of 95% ethanol eluate which are used as cyclocarya paliurus leaf extracts rich in dammarane triterpenoid saponin. Most of dammarane triterpenoid saponin compounds provided by the invention can inhibit alpha-glycosidase and inhibit release of nitric oxide in lipopolysaccharide-induced RAW264.7 cells, show good hypoglycemic and anti-inflammatory activity, have concentration dependence, and can be used for developing hypoglycemic and anti-inflammatory related drugs and health products.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a preparation method of dammarane-type saponins and its application in the preparation of hypoglycemic and anti-inflammatory drugs and health care products. Background technique [0002] Studies have shown that type 2 diabetes is related to cytokine-mediated inflammation, which is an innate immune disease. Inflammation plays a mediating role in the pathogenesis of type 2 diabetes. This view is called the "inflammation theory" of diabetes. At the American Diabetes Association meeting in 2005, this theory was widely recognized, and the relationship between "inflammation and type 2 diabetes" has become a common focus of global endocrinologists. [0003] During the onset of type 2 diabetes, the endogenous mRNA levels of NO, iNOS, NF-κB, COX-2, IL-6, IL-1β, and TNF-α in the patient's body increased significantly. It also inhibits the expression of NO, TNF-α, IL-6 and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07J17/00A61K31/58A61P3/10A61P29/00A23L33/105
CPCC07J17/005A61P3/10A61P29/00A23L33/105A23V2002/00A23V2200/30A23V2200/328A23V2250/21
Inventor 李俊李陈国黄锡山梁宏陈振锋邓胜平黄艳
Owner GUANGXI NORMAL UNIV
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