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Herba epimedii total flavonoid enzymatic product and preparing method and application thereof

An enzymatic hydrolysis product, the technology of Epimedium, applied in the field of medicine, can solve the problems of cumbersome separation and purification steps, the unfavorable rational application of traditional Chinese medicine resources, and the inability to fully utilize active ingredients, so as to reduce production costs, be suitable for industrial production, and facilitate recycling and repetition. The effect of reuse

Active Publication Date: 2017-02-22
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, simply extracting icariin from Epimedium medicinal material and performing enzymatic hydrolysis not only requires cumbersome separation and purification steps, but the effective components in Epimedium medicinal material cannot be fully utilized, which is not conducive to the rational application of traditional Chinese medicine resources.

Method used

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  • Herba epimedii total flavonoid enzymatic product and preparing method and application thereof
  • Herba epimedii total flavonoid enzymatic product and preparing method and application thereof
  • Herba epimedii total flavonoid enzymatic product and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] This example provides a preferred way of enzymatically hydrolyzing total flavonoids of Epimedium using the method of the present invention, and further specifically describes the enzymatic hydrolysis method of the present invention.

[0031] 1. Preparation of total flavonoids from Epimedium

[0032] Epimedium is extracted with 6-40 (m / v) times of 40%-90% ethanol by heat reflux, concentrated, and further purified by macroporous adsorption resin method to obtain total Epimedium with a mass content of not less than 60%. flavonoids.

[0033] The main components of the total flavonoids of Epimedium are icariin, epimedin A, epimedin B, epimedin C and baohuoside I, and icariin, epimedin A, epimedin B, The total mass content of Huodin C and Baohuoside I is not less than 30%.

[0034] 2. Preparation and characterization of amino mesoporous silica support

[0035] Immerse silica in toluene dissolved with 3-aminopropyltriethoxysilane (C=50mM), and stir magnetically at 800r·min -...

Embodiment 2

[0062] This example illustrates the effect of enzymatic hydrolysis of total flavonoids of Epimedium using other immobilized carriers.

[0063] This example compares the enzymatic hydrolysis effects of total flavonoids of Epimedium herba using barium alginate, microspheres and the amino mesoporous silicon-immobilized enzyme of Example 1. The results are shown in Table 2:

[0064] Table 2 Comparison of immobilized helicase prepared by different enzyme immobilization techniques

[0065]

[0066] It can be seen that the use of amino mesoporous silicon immobilized helicase has higher binding efficiency, better stability and higher conversion rate than other immobilized carriers.

Embodiment 3

[0068] This example illustrates the antitumor activity of the enzymatic hydrolyzate of the present invention.

[0069] Precisely weigh 2.65 mg of the total flavonoids of Epimedium and the free enzyme hydrolyzate and immobilized enzyme hydrolyzate obtained by the method of Example 1, respectively dissolve in 25 μL DMSO to prepare a mother liquor. Then the mother solution was diluted 100 times with RPMI-1640 incomplete medium solution, and then diluted to different dosage concentrations: 4.0, 17.0, 66.0, 265.0, 1060.0 μg·mL -1 . The MCF-7, A549, HepG2 and Hela cells in the logarithmic growth phase were inoculated in 96-well plates, and after 24 hours of culture, 20 μL of the drug solution to be tested was added. The final concentrations of the drug solution were 0.4, 1.7, 6.6, 26.5, and 106.0 μg mL -1 . Set up 6 duplicate wells for each sample, and set up a blank group of incomplete medium without drugs. After continuing to culture for 48 hours, MTT solution (5.0 mg·mL -1 ) ...

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Abstract

The invention relates to a herba epimedii total flavonoid enzymatic product. Herba epimedii total flavonoid is generated through immobilized helicase, and the enzymatic product has higher antineoplastic activity than the herba epimedii total flavonoid archetype and free enzyme enzymatic product and has high oral absorptivity. The invention further discloses a preparing method of the enzymatic product. The immobilized enzyme is used for enzymolysis of herba epimedii total flavonoid, selected helicase is high in combining efficiency with a fixed carrier and high in stability, the enzymolysis conversion rate is large, the immobilized enzyme can be used circularly, resources can be saved, and green production is achieved. The invention further relates to application of the enzymatic product in antitumous effect, and the herba epimedii total flavonoid enzymatic product obtained through the technical scheme has a remarkable anti-proliferative effect on the human lung cancer cell A549, liver cancer HepG2 cell, breast cancer MCF-7 cell, cervical cancer Hela cell and can be used for treating the lung cancer, liver cancer, breast cancer and cervical cancer.

Description

technical field [0001] The invention relates to an enzymatic hydrolysis product of total flavonoids of Epimedium. The product is produced by hydrolyzing total flavonoids of Epimedium through immobilized helicase. The invention also relates to the preparation method of the enzymatic hydrolysis product and its application in antitumor, belonging to the technical field of medicine. Background technique [0002] Epimedium, as a traditional kidney tonifying Chinese medicine, has been used for thousands of years among the people. It has the functions of tonifying kidney yang, strengthening muscles and bones, and dispelling rheumatism. [0003] Epimedium mainly contains flavonoids, mainly including flavonoid glycosides such as icariin, epimedin A, epimedin B, epimedin C and baohuoside I (for the structure see figure 1 ). Modern pharmacological studies have shown that Epimedium flavone glycosides have good biological activity, in addition to anti-osteoporosis activity, it also has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/296A61P35/00
CPCA61K36/296A61K2236/19A61K2236/333A61K2236/51A61K2236/55
Inventor 陈彦刘聪燕彭静
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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