Wood frog antibiotic peptides and preparation technology and its application in antiviral drug

An antimicrobial peptide and alkaline technology, which is applied in the fields of biochemistry and biopharmaceuticals, can solve the problems of insufficient research on skin active peptides, and achieve good antiviral effect

Active Publication Date: 2009-06-17
TONGHUA KANGYUAN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

my country has a long history of the application of amphibian drugs, but the research on its active ingredients and pharmacological properties mainly focuses on small organic molecules such as alkaloids, and there are not many studies on its skin active peptides

Method used

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  • Wood frog antibiotic peptides and preparation technology and its application in antiviral drug
  • Wood frog antibiotic peptides and preparation technology and its application in antiviral drug
  • Wood frog antibiotic peptides and preparation technology and its application in antiviral drug

Examples

Experimental program
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Effect test

Embodiment 1

[0035] (1) Preparation of Rana antimicrobial peptide:

[0036] (1) Weigh 20g of dried Rana frog skin and soak it in 200ml pH4.0 0.2mol / L acetic acid-acetate buffer solution for 24h.

[0037] (2) Centrifuge the obtained extract at 5000 r.p.m for 30 min, discard the precipitate, and keep the supernatant. The supernatant was boiled for 5 minutes, cooled, centrifuged at 5000 r.p.m for 30 minutes, and the supernatant was passed through a 30KDa membrane to obtain the antimicrobial peptide filtrate. Store at 4°C for later use.

[0038] (3) According to the basic principle of determining the relative molecular weight of protein by gel chromatography, V12 (MW1.3KDa), insulin β chain (MW3.3KDa), insulin (MW5.7KDa) and cytochrome C (MW13KDa) and pancreatic Protease inhibitor (MW2.01KDa) was used as a standard protein, mixed with the purified sample and dissolved, then applied to a Sephadex G-25 column, and the elution volume of each component was measured. Take Ve / V0 as the abscissa, ...

Embodiment 2

[0055] (1) Preparation of Rana antimicrobial peptide:

[0056] (1) Weigh 10 g of dried Rana frog skin and soak it in 120 ml of acetic acid-acetate buffer solution with pH 4.0 0.2 mol / L for 30 h.

[0057] (2) Centrifuge the obtained extract at 5000 r.p.m for 30 min, discard the precipitate, and keep the supernatant. The supernatant was boiled for 5 minutes, cooled, centrifuged at 5000 r.p.m for 30 minutes, and the supernatant was passed through a 30KDa membrane to obtain the antimicrobial peptide filtrate. Store at 4°C for later use.

[0058] (3) According to the basic principle of determining the relative molecular weight of protein by gel chromatography, V12 (MW1.3KDa), insulin β chain (MW3.3KDa), insulin (MW5.7KDa) and cytochrome C (MW13KDa) and pancreatic Protease inhibitor (MW2.01KDa) was used as a standard protein, mixed with the purified sample and dissolved, then applied to a Sephadex G-25 column, and the elution volume of each component was measured. Take Ve / V0 as t...

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Abstract

The invention provides a wood frog antibiotic peptide and a preparation process thereof. The invention further discloses application of the antibiotic peptide in the preparation of antiviral medicines. The peptide is a basic polypeptide with the molecular weight range from 2 to 16 KDa and the isoelectric point from 8.9 to 9.5. The peptide consists of asparagic acid, glutamic acid, major serine, histidine, arginine, glycine, threonine, proline, alanine, Val, methionine, and the like, wherein the basic amino acid accounts for 23.99 percent of the gross amount, and the acidic amino acid accounts for 19.08 percent of the gross amount. The wood frog antiviral antibiotic peptide is a product obtained from natural resources. The wood frog antiviral antibiotic peptide has the good antiviral capability and simultaneously has the advantages of no hemolytic activity, no plasma coagulation activity, and the like. The wood frog antibiotic peptide can be used for preparing medicines for treating antiviral type flu, hepatitis B and AIDS.

Description

Technical field: [0001] The invention provides a Rana antibacterial peptide and a preparation process of the antibacterial peptide. The invention further discloses the use of the antibacterial peptide in the preparation of antiviral drugs, which belongs to the technical field of biochemistry and biopharmaceuticals. Background technique: [0002] Many amphibians are widely used as traditional important and national medicines in China, such as Chinese Bufogargarizans, Bombina maxima, Pelophylax nigromaculata, etc. The skin and viscera of these amphibians have a wide range of pharmacological activities and clinical curative effects. Pharmacological activities have been reported to include spectrum antimicrobial effects, antitumor effects, analgesia, local anesthesia, immune regulation, and cardiovascular system effects. The search for specific pharmacologically active monomeric compounds in amphibian skin has become a hot spot for new drug discovery. The application of amphibi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/14A61K38/17A61P31/16A61P31/18A61P31/20
Inventor 滕利荣孟庆繁逯家辉安金双李青山周杰田晓乐高朝辉王贞佐
Owner TONGHUA KANGYUAN BIOLOGICAL TECH
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