Method for preparing active polypeptide from Arca subcrenata lischke

A technology of active peptides and cockles, applied in the preparation method of peptides, chemical instruments and methods, peptides, etc.

Inactive Publication Date: 2018-06-26
刘明明
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  • Abstract
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  • Application Information

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Problems solved by technology

The above patent documents describe in detail the extraction process and pharmacological activity of active substances in cockles, but more in-depth research on active substances, such as structural analysis and research on physical and chemical properties, has not been reported.

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  • Method for preparing active polypeptide from Arca subcrenata lischke

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[0008] The present invention will be further described below in conjunction with the examples.

[0009] Shell the cockles, take the cockle meat, wash with water at 4°C, homogenate, add distilled water at a low temperature (4°C) for extraction for 2 hours at a weight ratio of 1:1, flocculate, centrifuge to remove slag, take the supernatant, and After the supernatant was lyophilized, it was subjected to gel column chromatography with Sephadex G25 (0.03mol / 1NH 4 HCO 3 elution buffer), after collecting the peaks respectively, take the peaks with biological activity and then use anionic Sephadex DEAE-Sephadex A25 ion exchange column chromatography (PH8.5 Tris-HC1 buffer, 0 ~ 0.03mol / NaCl Gradient elution), the active peaks were collected, and after freeze-drying, desalted with Sephadex G15 gel column to obtain natural small molecule active peptides with high purity. The active peptide is a naturally extracted substance without any chemical modification, and the extraction process...

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Abstract

The invention relates to a method for extracting active polypeptide from Arca subcrenata lischke. The method comprises: removing shell from Arca subcrenata lischke, washing, homogenizing, adding distilled water, carrying out low temperature extraction, carrying out flocculation, carrying out centrifugation and residue removing, taking the supernatant, carrying out freeze drying on the supernatant,carrying out gel column chromatography, collecting each peak, carrying out ion exchange column chromatography to separate and purify, collecting the active peaks, carrying out freeze drying, desalting with a gel column to obtain natural active polypeptide with high purity, and continuously purifying the obtained small molecule peptide by using a C18 reverse phase column to give the active peptidewith a purity of more than 95%. According to the present invention, the molecular weight of the small molecule peptides obtained from Arca subcrenata lischke through the method is 500-3000 Da, and the biological activity is effectively protected during the extraction process; and the obtained active peptide has significant antitumor activity, and the in vitro antitumor experiment results show that the obtained active peptide has the tumor inhibition rate of 52%, and can be used for the development and application of novel antitumor drugs.

Description

Technical field: [0001] The invention belongs to the technical field of marine natural product extraction and medicine, and relates to a method for extracting small-molecule active peptides from cockles, in particular to a method for separating and purifying active peptides from cockles by column chromatography. Background technique: [0002] Scapharca subcrenata (Arca subcrenata Lischke; Scapharca subcrenata) belongs to the phylum Molluscs, Class Phyllobranches, Order Dendonta, and Scapharca family. It is one of the abundant marine organisms in my country and has a long history of medicinal use. Its shell is named corrugated fruit, which has been recorded in past materia medica. It has the functions of reducing phlegm, softening hardness, reducing acid and relieving pain. The cockle meat is a soft part, which is mostly contained in dietotherapy herbal medicines. The clam as a whole is used as medicine, which has the function of shell and meat. It can nourish the blood and w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/20C07K1/18C07K1/16A61P35/00
CPCC07K1/20C07K1/16C07K1/18
Inventor 尤世元孙亮刘明明
Owner 刘明明
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