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Centipede extract capable of resisting tumor activity and preparation method thereof

A technology of anti-tumor activity and centipede extract, which is applied in the field of centipede extract with anti-tumor activity and its preparation, and can solve the problems of non-testing of anti-tumor activity

Inactive Publication Date: 2011-07-27
HUNAN UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Or use the biomimetic enzymatic hydrolysis method to extract the effective parts of the centipede, after the enzymolysis is completed, put it in a constant temperature water bath at 85°C for 30 minutes to inactivate it, collect the supernatant and dry it to obtain the centipede enzymatic hydrolyzate, use the APTT method and the fibrinogen plate method as the inspection indicators, and compare the published The application of the extracted effective parts in anticoagulant and thrombolytic drugs (Chinese patent: CN101862349), the centipede hydrolyzate extracted by the process has not been further purified and separated, and the anti-tumor activity has not been tested

Method used

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  • Centipede extract capable of resisting tumor activity and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0032] Extraction of centipede anti-tumor small molecule active peptide components: Weigh 4 g of centipede micropowder, add 5 times the volume of ultrapure water, mix in an oscillating mixer after initial mixing to make a suspension. Add trypsin to the suspension to make the final concentration 0.5%, adjust the pH value to 7.0, keep the temperature at 4°C and hydrolyze under low temperature conditions, extract repeatedly 5 times, each time for 60 minutes, filter, combine the filtrates, and centrifuge at 3000g for 5 minutes Then collect the supernatant. The supernatant was taken and centrifuged at 5000 rpm for 10 minutes at 4°C. Take the supernatant, add twice the volume of pre-cooled acetone (this process is carried out on ice), shake, and centrifuge at 5000g for 10 minutes at 4°C. Take the precipitate, dissolve it in water, add twice the volume of pre-cooled acetone (this process is carried out on ice), shake, and centrifuge at 5000g for 10 minutes at 4°C. Take the precipit...

Embodiment 2

[0034] Proliferation inhibitory effect of centipede anti-tumor small molecule active peptide components on human liver cancer HepG2 cells: human liver cancer HepG2 cells in the logarithmic growth phase were digested to make a single-cell suspension, and the cell concentration was adjusted to 5×10 4 cells / ml, 200 μl cell suspension per well, inoculated in 96-well culture plate. 37°C, 5% CO 2 and saturated humidity incubator for 24h to adhere to the wall, divided into experimental group and control group. Centipede anti-tumor small molecule active peptide components are set to five gradient drug concentrations of 160 μg / ml, 80 μg / ml, 40 μg / ml, 20 μg / ml, and 10 μg / ml. At the same time, a negative control group is set to only add complete medium, and the well is adjusted to zero Add corresponding concentrations of drug working solution and complete medium, and set 5 replicate wells for each concentration in the experimental group and control group. After culturing at 37°C, 5% CO...

Embodiment 3

[0039] Inhibitory effect on the proliferation of human liver cancer MHCC97-H cells by centipede anti-tumor small molecule active peptide components: Human liver cancer MHCC97-H cells in the logarithmic growth phase were digested to make a single-cell suspension, and the cell concentration was adjusted to 5×10 4 cells / ml, 200 μl cell suspension per well, inoculated in 96-well culture plate. 37°C, 5% CO 2 and saturated humidity incubator for 24h to adhere to the wall, divided into experimental group and control group. Centipede anti-tumor small molecule active peptide components are set to five gradient drug concentrations of 160 μg / ml, 80 μg / ml, 40 μg / ml, 20 μg / ml, and 10 μg / ml. At the same time, a negative control group is set to only add complete medium, and the well is adjusted to zero Add corresponding concentrations of drug working solution and complete medium, and set 5 replicate wells for each concentration in the experimental group and control group. After culturing a...

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Abstract

The invention relates to an extract capable of resisting tumor activity, which is analyzed from centipede (a traditional Chinese medicine). The extract is obtained as follows: crushing the centipede at low temperature, conducting enzymolysis on the centipede at low temperature, separating and freeze-drying the centipede, and separating the centipede by a sephadex G-25 gel column, thereby obtaining an active ingredient which mainly comprises an effective ingredient, water-solubility micromolecule active peptide. By being tested by MTT, the extract has better anti-tumor activity. The extract avoids the defects that in the clinical application of a macromolecule protein medicine, certain allergenicity exists, and the quality control is not easy in industrialization caused by using the centipede (which is adopted as a crude drug), and has the advantages of a micromolecule activity peptide medicine, including high production efficiency, work procedure saving, quality control convenience and the like.

Description

technical field [0001] The invention belongs to a centipede extract, in particular to a centipede extract with antitumor activity and a preparation method thereof. Background technique [0002] Centipede is the dry body of centipede with few spiny giant centipedes. It is mainly produced in Jingzhou, Yichang, Xiaogan, Yunyang and other cities in Hubei, as well as Laohekou, Xiangfan, Jingmen, Zaoyang and other places. Warm in nature, pungent in taste, poisonous. Return liver, spleen, lung meridian. It has the functions of relieving wind and relieving spasm, attacking poison and dispelling stagnation, dredging collaterals and relieving pain. It is used for convulsions in children, convulsions, crooked mouth and eyes due to apoplexy, hemiplegia, tetanus, rheumatism, sores, scrofula, and snake bites. "Compendium of Materia Medica" records that centipedes can be used for erysipelas tumors, children's mouth, children's panic, convulsions, tetanus, crooked mouth and eyes, numbne...

Claims

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Application Information

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IPC IPC(8): A61K35/64A61P35/00A61K35/648
Inventor 田雪飞周青廖兴华孙婧艾小佳
Owner HUNAN UNIV OF CHINESE MEDICINE
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