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Active material of polypeptide proteins in bloody clam, preparation method and usage

A technology of active substances and peptide proteins, applied in the field of cockles, can solve the problem that the extract cannot be used directly

Inactive Publication Date: 2005-03-30
于荣敏 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The resulting extract cannot be used directly in applications requiring high purity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Take cockles, remove their shells, take their contents, wash them with normal water at 4°C for 3 times, and then wash with distilled water at 4°C twice; add distilled water at a weight ratio of 1:3, homogenate at 5000r / min for 3min, Remove the slag on the centrifuge, get the supernatant after centrifuging for 10min; carry out ion exchange column chromatography to the supernatant (using DEAE anion exchanger, adopting phosphate buffer solution to carry out 0.8-1.5M salt concentration gradient elution), by this step, the The extract is separated and purified according to the difference in the adsorption capacity of the ion exchange medium. After each peak was collected separately, it was desalted by dialysis. After desalting, it is separated by gel column chromatography (the filler is Sephadex G-50, and eluted with phosphate buffer), and the extract with the same adsorption force is further separated according to different molecular weights, and the obtained molecular weig...

Embodiment 2

[0014] The polypeptide and protein active substances of the cockles above were used for in vivo tumor experiment on tumor-bearing mice. Male Kunming mice weighing 19-21 g were randomly divided into groups of 10 mice. Each mouse was inoculated with 0.2ml S in the left armpit 180 Tumor fluid of Ehrlich ascites carcinoma. The next day, normal saline was used as a negative control and 5-fluorouracil was used as a positive control for intraperitoneal injection. Eight days after the injection, the animals were weighed, and the subcutaneous tumor mass was dissected and weighed. The curative effect of solid tumors is expressed by the percentage of inhibition of tumor weight, and the calculation formula is as follows:

[0015] Tumor weight inhibition rate%=(1-T / C)×100%

[0016] T: average tumor weight in the treatment group

[0017] C: average tumor weight in the negative control group

[0018] drug on mouse S 180 Inhibition of tumor

[0019] Group Ad...

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Abstract

The invention relates to multipiptide protein active substance in a kind of bloody clam (Arca subcrenata Lischke; Scapharca subcrenata), the prepn. method and the uses. The prepn. method is that bloody clam is homogenated to obtain superantant which is treated by chromatography in ion exchange column, desalt, chromatograph separation in gel colum to be combined to several peaks with molecular weight 3-15 KDa which is concentrated and dried to obtain multipeptide protein active substance. Said active substance has 73.72 percent of cancer-inhibiting ratio and can be made as anti-cancer medicine.

Description

Technical field: [0001] The invention discloses a polypeptide protein active substance in clams (Arca subcrenata Lischke; Scapharca subcrenata;), a preparation method and application thereof. Background technique: [0002] Hairy clams (Arca subcrenata Lischke; Scapharca subcrenata;) belong to the phylum Molluscs, Phyllobranches, Order Dendontia, and Scapharca family. Its shell is Chinese medicine corrugated seed. There are 31 to 34 radial ribs. The surface of the shell is hairy and brown in color. Produced in the northern coast of my country. [0003] "China Marine Medicine" magazine, 2003, No. 2 P37, introduced the separation method of oyster protein. Fresh oyster soft body, homogenate with 2 times the amount of normal saline at 4°C, centrifuge at 3000r / min for 15min, take the supernatant, add (NH) 2 SO 4 , so that the solution's (NH) 2 SO 4 The saturations reached 50%, 75% and 100%, respectively, and stood at 4°C for 12h. Then centrifuge at 3000r / min for 15min, ta...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P35/00C07K1/22C07K14/435
Inventor 于荣敏赵昱
Owner 于荣敏
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