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A kind of enzymatic hydrolysis product of epimedium total flavonoids and its preparation method and application

An enzymatic hydrolysis product, the technology of Epimedium, applied in the field of medicine, can solve the problems of unfavorable traditional Chinese medicine resources rational application, complicated separation and purification steps, insufficient utilization of effective components, etc., and achieves suitable industrial production, reduces production costs, and facilitates recovery and repetition. The effect of reuse

Active Publication Date: 2019-04-30
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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  • A kind of enzymatic hydrolysis product of epimedium total flavonoids and its preparation method and application
  • A kind of enzymatic hydrolysis product of epimedium total flavonoids and its preparation method and application
  • A kind of enzymatic hydrolysis product of epimedium total flavonoids and its preparation method and application

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 an enzymatic hydrolysis product of total flavonoids of Epimedium, which is produced by enzymatic hydrolysis of total flavonoids of Epimedium through immobilized snail enzymes, and the enzymatic hydrolysis product has higher Better anti-tumor activity and better oral absorption. The invention also discloses a preparation method of the enzymatic hydrolysis product. The method uses immobilized enzymes to enzymolyze the total flavonoids of Epimedium herba, and the selected helicase has high binding efficiency, good stability, and high enzymatic hydrolysis conversion rate with the immobilized carrier. Moreover, the immobilized enzyme can also be recycled, which can save resources and realize green production. The present invention also relates to the application of the enzymatic hydrolyzate in anti-tumor. The total flavonoid enzymatic hydrolyzate of Epimedium herba obtained according to the technical scheme provided by the present invention is effective against human lung cancer A549 cells, liver cancer HepG2 cells, breast cancer MCF‑7 cells, cervical cancer Hela cells have significant proliferation inhibitory effect and can be used for the treatment of 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 Patents(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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