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Three alpha-conus polypeptides and use thereof

A sequence and amino acid technology, applied in the field of α-type conus polypeptide, can solve problems such as tolerance of toxic side effects and poor therapeutic effect

Inactive Publication Date: 2013-06-19
INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically commonly used drugs mainly include the combination of anticonvulsants (such as leticitide and oxcarbazepine), opioids (such as morphine) and various antidepressants. Receptivity, addiction and other problems, the treatment effect is not good (Dray et al, Br J Anaesth.2008, 101(1), 48-58; Gilron et al, Expert Opin Emerg Drugs.2007, 12(1), 113 -26.)

Method used

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  • Three alpha-conus polypeptides and use thereof
  • Three alpha-conus polypeptides and use thereof
  • Three alpha-conus polypeptides and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Embodiment 1, the discovery of α-type conus polypeptide

[0082] 1. Extraction of cone snail total RNA

[0083] Six kinds of live snails were collected from the Xisha Islands and Sanya, Hainan as sample materials, and the total RNA of the six kinds of snails was extracted by Trizol method. 6 kinds of cone snails: marble cone (Conus marmoreus), stately cone (Conus imperialis), signal cone (Conus litteratus), black heart cone (Conus emaciatus), maiden cone (Conus eberneus), calf cone (Conus Conus vitulinus).

[0084] The specific steps of Trizol method to extract total RNA:

[0085] ①Take the cone venom tube on ice first, freeze it in liquid nitrogen, grind it into powder with a grinder, and pour it into a micro homogenizer that has been ice-bathed in advance.

[0086] ②Pipe 1.2ml Trizol into the homogenizer and grind further to lyse the cone snail tissue to release RNA.

[0087] ③ Pour the ground slurry into a 1.5ml centrifuge tube and let stand at room temperature for...

Embodiment 2

[0115] The synthesis of embodiment 2, alpha type conus polypeptide

[0116] The seven polypeptides shown in Table 3 were synthesized respectively.

[0117] Amino acid sequence of table 3α type conus polypeptide

[0118] name

sequence

T-1

NH 2 -GCCSNPACMLKNPNLC*

T-2

NH 2 -NCCTRSFCKRIYPDLC*

T-3

NH 2 -GCCDIPDCYNKNREQC*

T-4

NH 2 -NCCMFHTCPIDYSRFNC*

T-5

NH 2 -GNCCMFHTCPIDYSRFNC*

T-6

NH 2 -GNCCMFHTCPIDYSRFYC*

T-7

NH 2 -RKCCSNPACNRYNKLC

[0119] D-aspartic acid; E-glutamic acid; I-isoleucine; K-lysine; L-leucine; M-methionine; N-asparagine; Q-glutamine; R- Arginine; S-serine; T-threonine; G-glycine; F-phenylalanine; P-proline; H-histidine; A-alanine; Y-tyrosine. For T-1 and T-7, the disulfide linkages in each polypeptide are C1-C3, C2-C4. * indicates amidation (CONH 2 ).

[0120] 1. Peptides

[0121] (1) Synthesis of T-1 (artificially synthesized by Fmoc ...

Embodiment 3

[0172] Embodiment 3, the analgesic activity assay of α-type conus polypeptide

[0173] 1. Preparation of α-cono polypeptide Vc1.1

[0174] α-cono polypeptide Vc1.1: GCCSDPRCNYDHPEICONH2.

[0175] 1. Synthesis of peptide resin

[0176] using solid-phase synthesis techniques. It was carried out on a 433A polypeptide synthesizer (ABI Applied Systems Biological Instruments). The amino acid used in the synthesis is Fmoc protected amino acid (product of Advanced Chemtech Company in the United States), the resin used is Rink resin (product of Advanced Chemtech Company in the United States), and the reagents used include DCC (Acros Company), HOBt (product of Advanced Chemtech Company in the United States), NMP (PE company), piperidine (Shanghai Gil Biochemical), methanol (domestic analytical grade) and dichloromethane (domestic analytical grade).

[0177] The side chain protecting groups of Fmoc amino acids are as follows: the side chain protecting groups of His and Cys are trityl...

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Abstract

The invention discloses alpha-conus polypeptides and use thereof. The invention provides seven polypeptides: a polypeptide I of which the amino acid sequence is represented by a sequence 7 in a sequence table and in which the last amino acid residue is amidated; a polypeptide II of which the amino acid sequence is represented by a sequence 8 in the sequence table and in which the last amino acid residue is amidated; a polypeptide III of which the amino acid sequence is represented by a sequence 9 in the sequence table and in which the last amino acid residue is amidated; a polypeptide IV of which the amino acid sequence is represented by a sequence 10 in the sequence table and in which the last amino acid residue is amidated; a polypeptide V of which the amino acid sequence is representedby a sequence 11 in the sequence table and in which the last amino acid residue is amidated; a polypeptide vI of which the amino acid sequence is represented by a sequence 12in the sequence table andin which the last amino acid residue is amidated; and a polypeptide VII of which the amino acid sequence is represented by a sequence 13 in the sequence table and in which the last amino acid residueis amidated. The result of activity tests, the seven polypeptides of the general formula (I) have pain relieving effect, alpha-conus polypeptides T-1, T-2, T-3 and T-4 have obvious pain-relieving activity, and the alpha-conus polypeptides T-5, T-6 and T-7 also have obvious pain-relieving effect, and the alpha-conus polypeptides have application values in pain-reliving aspect.

Description

[0001] This application is a divisional application of the patent application with the application number 201010121879.7, the application date is March 10, 2010, and the invention title is "α-type conus polypeptide and its application". technical field [0002] The invention relates to an alpha-type conus polypeptide and its application. Background technique [0003] Cono snails belong to gastropod molluscs, and there are more than 500 species in the world, all over the world's warm sea areas. There are more than 100 species of cone snails in China, mainly distributed in my country's Xisha Islands, Hainan Island and Taiwan waters. Cono snail polypeptides are secreted by the venom glands of the cone snail venom tube and the inner wall of the poison sac. The venom of each cone snail contains 50-200 active peptides. Different species of cone snails contain different active peptides. Even the same species of taro Cone venom-from accidental stings to deliberate injection. Toxicon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C12N15/12A61K38/10A61P29/00
Inventor 刘珠果戴秋云孙婷吴巧玲胡洁于正
Owner INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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