Acidian polypeptide and preparation method thereof

A technology of sea squirt polypeptide and sea squirt, which is applied in the field of sea squirt polypeptide and its preparation, and can solve problems such as applications that have not been reported yet

Inactive Publication Date: 2006-07-19
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the patent application of sea squirts has been reported on sea squirts and fatty acids, but the preparation method of sea squirt polypeptides extracted from sea squirts and its application in the treatment of hepatitis B have not yet been reported.

Method used

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  • Acidian polypeptide and preparation method thereof
  • Acidian polypeptide and preparation method thereof
  • Acidian polypeptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 50Kg of fresh sea squirts, eviscerated, washed, soaked with 50kg of 75% ethanol at room temperature for 2 weeks, recovered alcohol with a rotary evaporator to obtain 1000g of extract. Dissolve 200g of the extract in 500ml of distilled water, extract with the same volume of dichloromethane, dry up the solvent to obtain 825g of the water-soluble part; then dissolve as much as possible in 500ml of distilled water, extract with the same volume of n-butanol, dry up the solvent to get the water-soluble part 712g; then take 100g of the water-soluble part, put it on D-101 macroporous resin column chromatography, elute with 500ml of 50% ethanol, the flow rate is 3ml / min, and dry the solvent to obtain 14g of product; carry out silica gel column chromatography with CHCl 3 :MeOH:H 2 O ratio is 7:3:0.5 eluent gradient elution, flow rate is 3ml / min, collect thin-layer chromatography ultraviolet light to detect R f The eluent is 0.2-0.3, and the solvent is dried to obtain the crude e...

Embodiment 2

[0047] Take 50 kg of fresh Ascidian rugosa, remove the viscera, wash, soak in 50 kg of 75% ethanol at room temperature for 2 weeks, recover the alcohol with a rotary evaporator at 60 ° C until it is odorless, and obtain 1000 g of extract. Take 200g of the extract and dissolve it with 500ml of distilled water as much as possible, extract with the same volume of chloroform, dry up the solvent to obtain 805g of the water-soluble part; then dissolve it with 500ml of distilled water as much as possible, extract with the same volume of n-butanol, and dry up the solvent to get 683g of the water-soluble part Then take 100g of the water-soluble part, put it on AB-8 macroporous resin column chromatography, elute with 500ml of 50% ethanol, the flow rate is 2.5ml / min, and dry the solvent to obtain 10g of product; carry out silica gel column chromatography with CHCl 3 :MeOH:H 2 O ratio is 7:3:0.5 eluent gradient elution, flow rate is 2.5ml / min, collect thin-layer chromatography ultraviolet...

Embodiment 3

[0049] 50 kg of fresh Ascidian rugosa, eviscerated, washed, soaked in 50 kg of 90% ethanol at room temperature for 2 weeks, recovered the alcohol with a rotary evaporator at 60 ° C until it was odorless, and obtained 1300 g of extract. Take 200g of the extract and dissolve it in 600ml of distilled water, extract with the same volume of chloroform, dry up the solvent to obtain 830g of water-soluble part; repeat the above extraction method 3 times, dry up the solvent to get 730g of water-soluble part; then take 100g of water-soluble part, put on HDP- 100 macroporous resin column chromatography, elute with 500ml of 50% ethanol, the flow rate is 2ml / min, and the solvent is collected to obtain 15g of product; carry out silica gel column chromatography with CHCl 3 :MeOH:H 2 O ratio is 7: 3: 0.5 eluent gradient elution, flow rate is 3ml / min, collects the eluent that thin-layer chromatography ultraviolet light detection Rf is 0.2-0.3, collects dry solvent and obtains ascidian polypept...

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Abstract

The low-cost preparation method for polypeptide sea squirts with sequence as formula (I), 666.3 molecular weight and anti-HBV function comprises: dipping with ethanol at normal temperature to prepare extract after ethanol recovery; extracting the extract with organic solvent to condense, separate and purify by in turn the chromatography with macroporous resin, silica gel with gradient elution with CHCl3, MeOH and H2O by ratio as 7:3:0.5, and Sephadex LH20; collecting and condensing the eluent with Rf as 0.2-0.3 for ultraviolet detection label in TLC.

Description

field of invention [0001] The invention belongs to the field of biochemistry and biomedicine, and specifically relates to ascidian polypeptide and a preparation method thereof. Background technique [0002] Ascidia is a Uronordata subphylum Uronordata, and a few of the anticancer components extracted from Ascidia have entered the clinical or preclinical stage. Studies have found that sea squirts contain dozens of compounds such as alkaloids, peptides, indoles, heavy metal chelating agents, polysulfides, macrolides, and terpenes, which have strong antitumor, antiviral, and antibacterial properties. , Inducing sarcoplasmic reticulum to release calcium, inhibiting calmodulin activity and other physiological activities, especially anti-tumor research reports (Li Zhijun, Xue Changhu, Wang Changhai. Anti-tumor bioactive substances in sea squirts. Ocean Bulletin, 2004, 23(5):82-86; Ji Yubin, Chi Wenjie. Research on anti-tumor active substances in sea squirts. Journal of Harbin Uni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/14
Inventor 万新祥胡文军曾凡林王瑞
Owner SOUTHERN MEDICAL UNIVERSITY
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