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Separation and purification method of active starfish peptide, active starfish peptide and application

A sea star, active technology, applied in the field of sea star peptide separation and purification, can solve the problems of large toxic and side reactions, poor selectivity, toxic and side effects, and achieve the effects of easy operation, strong targeting and good biocompatibility

Active Publication Date: 2021-12-24
渤海水产股份有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The main method of clinical treatment of breast cancer is surgery combined with radiotherapy and chemotherapy. However, surgical treatment is prone to recurrence, and radiotherapy and chemotherapy have strong toxic and side effects. Although chemotherapy drugs have certain curative effects on most tumors, they have poor selectivity and toxic side effects. Problems such as large scale and drug resistance are still very obvious

Method used

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  • Separation and purification method of active starfish peptide, active starfish peptide and application
  • Separation and purification method of active starfish peptide, active starfish peptide and application
  • Separation and purification method of active starfish peptide, active starfish peptide and application

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Embodiment approach

[0050] As a preferred embodiment of the present invention, the separation and purification method of active starfish peptide comprises the following steps:

[0051] (a) Dissolve the crude starfish peptide in pure water, filter to obtain a starfish peptide sample, and refrigerate at -4°C for later use;

[0052](b) Put the swollen chromatography filler (including absolute ethanol in the preservation liquid), after standing and settling, discard the upper layer liquid and the floating chromatography filler, add an appropriate amount of ultrapure water, and repeat washing until the ethanol is completely remove;

[0053] Connect the AKTA instrument, column and column packer, shake the chromatography filler evenly and pour it into the column packer quickly, after natural settlement for 24 hours, press the column with a mobile phase at a certain flow rate until the interface of the filler in the column no longer moves down , use BSA to detect column efficiency to check the packing e...

Embodiment 1

[0075] This embodiment provides a method for separation and purification of active starfish peptide, comprising the following steps:

[0076] (a) Wash the starfish with clean water, remove the viscera, cut it into pieces, discard as much water as possible, and weigh it. Add ultrapure water twice its mass to the shredded starfish body wall. Homogenize with a tissue homogenizer (22000rpm, 10min) to obtain starfish homogenate. Add 2 times the volume of PBS of the homogenate and a compound enzyme of 1% starfish mass (the compound enzyme is alkaline protease, papain, and trypsin prepared at an activity unit ratio of 3:4:4), and put the above homogenate at 50°C Enzymolysis was carried out in a constant temperature water bath for 10 hours, stirring continuously, and keeping the pH between 6.8 and 7.2.

[0077] After the enzymatic hydrolysis, three times the volume of 95% ethanol was added to the enzymatic hydrolysis solution, and it was placed in a refrigerator at 4°C for 6 hours. ...

experiment example 1

[0087] Experimental Example 1 Inhibition of proliferation of breast cancer cell MCF-7

[0088] (1) Crude starfish peptide

[0089] Accurately weigh 5 mg of the crude starfish peptide of Example 1, and dissolve it in 5 mL of DMEM medium containing 10% FBS (fetal bovine serum), wherein the final concentration of the starfish peptide crude product is 1 mg / mL. After being sterilized by filtration through a 0.22μm microporous membrane, it was used as the mother solution for anti-tumor activity test, and the other concentrations used were diluted from this concentration. Breast cancer cells MCF-7 were plated and cultured for 24 hours until they adhered to the wall. The starfish peptide anti-tumor activity test mother solution was diluted to 100 μg / mL, 200 μg / mL, 300 μg / mL, 400 μg / mL, and 500 μg / mL respectively. Six replicate wells were set up for each concentration. Aspirate the medium in the wells before adding the drug, and add 100 μL of the starfish peptide crude product soluti...

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Abstract

The invention provides a method for separating and purifying an active starfish peptide, the active starfish peptide and its application, and relates to the technical field of separation and purification of starfish peptides. In the separation and purification method, the crude starfish peptide is purified and separated by gel chromatography column chromatography to obtain the active starfish peptide; the separation and purification method is simple and easy to operate, and the preparation process is safe and environmentally friendly, which enriches the knowledge of active substances in starfish. The research provides a theoretical basis and experimental basis for promoting the application of starfish resources. The present invention also provides the active starfish peptide prepared by the above separation and purification method. The active starfish peptide has the advantages of high purity, high activity, good biocompatibility, safety and no side effects, and has good application prospects. The present invention also provides the application of the above-mentioned active starfish peptide in the preparation of a drug for inhibiting the proliferation of breast cancer cell MCF-7 or in the preparation of an anti-breast cancer drug.

Description

technical field [0001] The invention relates to the technical field of separation and purification of starfish peptides, in particular to a method for separation and purification of active starfish peptides, active starfish peptides and applications. Background technique [0002] Asterias rollestoni (Asterias rollestoni), also known as starfish, belongs to the family Echinodermata, Asteria, and Acanthidae. my country's Weihai area is rich in aquatic resources, and the sea star production in coastal areas is large, but there is no effective development and utilization of starfish resources. The main active substances in starfish include starfish polypeptide, starfish polysaccharide and starfish saponin, etc., which are rich in medicinal value. Some polysaccharides in starfish have strong antibacterial and antiviral effects, and starfish total saponins have hemolytic effect. Bread starfish saponins-1 (astemsaponins-1) can significantly inhibit the growth of malignant glioma ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K1/16C07K1/34C07K1/36A61K38/01A61P35/00A61P15/14
CPCC12P21/06C07K1/16C07K1/34C07K1/36A61P35/00A61P15/14A61K38/00
Inventor 陈锚宋淑亮魏强丁茂昌关浩武心华
Owner 渤海水产股份有限公司
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