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Inoscavin A as a monomeric component in phellinus as well as prepearation method and application thereof

A technology of inoscavin and Phellinus, applied in the field of inoscavin A (inoscavin A) and its preparation, monomer components, can solve the problem of insufficient monomer components, save solvent processing time, and simplify the separation process , high separation efficiency

Active Publication Date: 2013-03-20
ZHEJIANG ACAD OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the technical problem that there is no legal and commercial standard substance for Phellinus at present, and the insufficient technical problems of monomer component preparation research, the first object of the present invention is to provide the monomer component in Phellinus radix-celloporin A compound, the compound Through in vitro experiments, it has the activity of inhibiting the proliferation of lung cancer tumor cells and liver cancer tumor cells; in vivo activity experiments have verified that it has anti-inflammatory, immune regulation and tumor chemotherapy drug synergistic effects, and provides materials for further pharmacodynamic research and quality evaluation and control of Phellinus Phellinus foundation, and ultimately provide new health food or drug options for health care and even disease treatment

Method used

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  • Inoscavin A as a monomeric component in phellinus as well as prepearation method and application thereof
  • Inoscavin A as a monomeric component in phellinus as well as prepearation method and application thereof
  • Inoscavin A as a monomeric component in phellinus as well as prepearation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A method for preparing and purifying a cellulosin A compound, the method comprising the following steps:

[0045] 1) Take Phellinus herb, crush it into large broad bean granules, weigh 1000g precisely, add 8000mL of 75% ethanol aqueous solution, soak for 3h, soak in a decoction machine at 80°C for 2h, filter; add 6000mL of quality to the filter residue Concentration of 75% ethanol aqueous solution, 80 ℃ warm immersion extraction for 1 hour, filter; add 4000mL mass concentration of 75% ethanol aqueous solution to the filter residue again, 80 ℃ warm immersion extraction for 0.5h, filter; combine 3 filtrates, concentrate under reduced pressure To a thick liquid with a volume of 210mL, the relative density measured at 40°C is 1.17, and it is set aside;

[0046] 2) Take 60mL of the thick liquid in step 1), add 600mL of hot water to dissolve, transfer to a separatory funnel, let cool; add 600mL of petroleum ether, shake and extract; let stand to separate layers; separate the ...

Embodiment 2

[0053] Example 2 Application of Phellinus Phellinus Medicinal Material Quality Evaluation

[0054] 1) Take the monomer component prepared by the present invention-celloporin A, add methanol to prepare a solution with a mass concentration of 0.1 mg / mL, and set aside;

[0055] 2) Take the coarse powder of Phellinus Phellinus medicinal material sample from each main production area of ​​the country, accurately weigh it, add ethanol solution with a mass concentration of 95%, soak for 1 hour, ultrasonically extract for 0.5 hour, filter the extract with a 0.45μ microporous membrane, and the filtrate As the test solution, standby;

[0056] 3) Use analytical high-performance liquid chromatography with a C18 column (4.6mm×250mm, 5μm) at a column temperature of 30°C, using methanol (phase A) and 0.2% phosphoric acid aqueous solution (phase B) as mobile phases. Gradient elution program: 0-20min, A phase 30-40%; 20-30min, A phase 30-40%; 30-45min, A phase 45%; 45-60min, A phase 45-55%, f...

Embodiment 3

[0058] Example 3 Study on the anti-lung adenocarcinoma (A549) cell effect of the monomer component fibroporin A in vitro

[0059] 1) Take celloporin A, a monomer component, and add dimethyl sulfoxide to prepare solutions with mass concentrations of 12.5, 25, 50, 100 and 200 μg / mL, respectively, and set aside;

[0060] 2) Take the human lung adenocarcinoma cell line (A549) in the logarithmic growth phase, use 1640 culture medium containing 10% fetal bovine serum to make 100uL single-cell suspension, and add it to a 96-well flat-bottomed culture plate Medium, 37℃, 5%CO 2 Cultivate in the incubator for 1 day and set aside;

[0061] 3) Take 100uL of solutions with different mass concentrations in step 1) and add them to the cell culture plate in step 2) respectively. Make 2 replicate wells for each drug concentration, and make 10% fetal bovine serum culture solution and The 5-fluorouracil (5-Fu) control with a mass concentration of 25ug / mL was cultured in a 37°C, 5% CO2 incubato...

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Abstract

The invention belongs to the field of medicaments and particularly relates to inoscavin A as a monomeric component in the phellinus as well as a preparation method and an application of the inoscavin A. The preparation method of the inoscavin A comprises the following steps of: (1) grinding the phellinus serving as a herbal medicine, and carrying out reflux extraction by ethanol; (2) extracting the ethanol extract by petroleum ether, and removing the petroleum ether; (3) extracting the extract by methylene dichloride, taking the ethylene dichloride layer, and drying to obtain the methylene dichloride extract; and (4) dissolving the extract into a flowing phase, carrying out high-speed counter-current chromatography, receiving and merging the flows, and recovering the solvent to obtain dried powder. Due to the adoption of the technical scheme, the compound of the inoscavin A as a monomeric component in the phellinus can be prepared in batch. The experiments in vitro prove that the compound has lung cancer tumor cell inhibition activity and liver cancer tumor cell proliferation activity. The experiments in vivo prove that the compound has the efficacy of diminishing inflammation and regulating immunity and can take effect together with the medicament for tumor chemotherapy so that the medicinal effect of the compound in vitro and vivo can be studied further.

Description

technical field [0001] The invention relates to a kind of monomer component extracted from Phellinus Phellinus, which belongs to the field of medicine: celloporin A (inoscavin A) And its preparation method and application. Background technique [0002] Cancer is the second leading cause of death in humans after cardiovascular disease, posing a serious threat to human health. It is estimated that by 2030, there will be about 21 million new cancer patients worldwide, and about 13.2 million deaths. There are about 2.2 million new cancer cases and 1.6 million deaths in my country every year. Therefore, the prevention and research of malignant tumors has become a research field that medical workers around the world pay close attention to. [0003] Chinese medicine treatment of cancer, because of its multi-target, multi-link and multi-effect characteristics, can improve the body's immunity and play an important role in prolonging the survival period and improving the quality of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D493/20A61P35/00A61P29/00A61P37/02A23L1/29G01N30/02A23L33/00
Inventor 董宇寿旦李洪玉俞忠明陈平华王绪平陈莉君黄孝闻章建民张扬吴人杰
Owner ZHEJIANG ACAD OF TRADITIONAL CHINESE MEDICINE
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