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Two delta-conotoxin polypeptides and a preparation method thereof

A conotoxin, polypeptide sequence technology, applied in the fields of biochemistry and molecular biology, to achieve the effect of strong analgesic activity

Active Publication Date: 2020-06-23
中国人民解放军军事科学院防化研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problem of the preparation of δ-conotoxin polypeptide and its application in the preparation of analgesic drugs, and provides two kinds of δ-conotoxin polypeptides and their preparation methods

Method used

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  • Two delta-conotoxin polypeptides and a preparation method thereof
  • Two delta-conotoxin polypeptides and a preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Separation and preparation method of δ-conotoxin polypeptide SEQ ID NO.1 and SEQ ID NO.2

[0020] Two kinds of isolated delta-conotoxin polypeptide sequences of the present invention are:

[0021] SEQ ID NO. 1: DECYSOGTFCGIKOGLCCSARCFSFVCISLEF;

[0022] SEQ ID NO. 2: DECFSOGTFCGIKOGLCCSAWCYSFFCLTLNF;

[0023] O is hydroxyproline, where the first cysteine ​​at the N-terminus forms a disulfide bond with the fourth cystine, and the second cysteine ​​forms a disulfide bond with the fifth cystine, And the third cysteine ​​forms a disulfide bond with the sixth cystine.

[0024] The steps of the preparation method for separating the conotoxin polypeptide SEQ ID NO.1 and SEQ ID NO.2 of the present invention from the cono venom duct of the agate are as follows:

[0025] (1) The dissected venom tube is 7 cm long, 0.5 cm in diameter, 0.5 cm long in radula, with barbs, and the venom bubble is very large, 1.5 cm long. Use alcohol-sterilized scissors to cut the thin thread-shaped...

Embodiment 2

[0030] In vivo analgesic experiment of two kinds of δ-conotoxin polypeptides in rats

[0031] In this example, an in vivo analgesic experiment in rats was conducted for the two δ-conotoxin polypeptides described in the present invention.

[0032] Experimental animals: male SD rats, SPF grade, 6-8 weeks old, weighing 180±20g, purchased from the Experimental Animal Center of the Chinese Academy of Military Medical Sciences, certificate number: SCXK-(Army) 2012-2004. Normal adaptive feeding conditions: rearing in independent isolation cages, temperature 18-21°C, humidity > 40%, 150Pa pressurized air supply, cycle light (12h light, 12h dark), free access to water and food (standard pellet feed), separated Change the bedding and feed every day.

[0033] Hot plate method experiment grouping and treatment: put the experimental rats on a hot plate, adjust the temperature to 55±0.5°C, and measure the normal pain response of each rat. Taking rats licking their hind paws or lifting the...

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Abstract

The invention discloses two delta-conotoxin polypeptides and a preparation method thereof, and belongs to the field of biochemistry and molecular biology. The amino acid sequence of the polypeptides is a brand new sequence, and the two polypeptides are prepared by adopting a conotoxin glandular tube separation and purification method of Agate. The two delta-conotoxin polypeptides are proved to have strong analgesic activity after being subjected to animal-level hot plate method and tail scalding method experiments. The two delta-conotoxin polypeptides are proved to have strong analgesic activity after being subjected to animal-level hot plate method and tail scalding method experiments, and can be applied to analgesic drugs.

Description

technical field [0001] The invention relates to two delta-conotoxin polypeptides and a preparation method, belonging to the fields of biochemistry and molecular biology. [0002] technical background [0003] Conotoxin is a kind of neurotoxin peptide secreted by cone snails and used to anesthetize prey. Due to its novel structure, strong activity and high selectivity, it can distinguish different subtypes of various ion channels, which opens up new opportunities for the study of ion channels. ideas. Conotoxins have strong biological activity and can specifically act on receptors and ion channels on animal cell membranes. Especially for voltage-gated or ligand-gated ion channels with high selectivity. Conotoxins have been widely used in neuropharmacology, and have excellent application prospects in analgesia, smoking cessation, drug abstinence, and the treatment of major diseases such as Parkinson's disease, neurodegenerative diseases, and mental illness. As a natural produ...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/14
CPCC07K14/43504
Inventor 蒋辉刘秀洁王康姚戈刘艳丽宋天宇万秀坤熊珊珊梁雅婷
Owner 中国人民解放军军事科学院防化研究院