Three alpha-conus polypeptides and use thereof

A linear polypeptide and sequence technology, applied in the application, peptide, peptide/protein components, etc., can solve the problems of toxicity and side effect tolerance, poor treatment effect, etc.

Inactive Publication Date: 2012-01-04
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] Example 1. Discovery of α-type Cono polypeptide

[0082] 1. Extraction of Total RNA from Cono

[0083] Six species of Cono snails were collected from the Paracel Islands, Sanya, Hainan and other places as sample materials, and the total RNA of the six species of Cono snails was extracted by the Trizol method. 6 types of Conus: Conus marmoreus (Conus marmoreus), Conus imperialis (Conus imperialis), Conus litteratus (Conus litteratus), Conus emaciatus (Conus emaciatus), Conus eberneus, Conus (conus) Conus vitulinus).

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

[0085] ① Take the cono venom gland tube on ice, freeze it in liquid nitrogen and grind it into powder with a grinder, and pour it into a micro-homogenizer in an ice bath.

[0086] ②Pick 1.2ml Trizol into a homogenizer and further grind it to lyse the cono tissue to release RNA.

[0087] ③Pour the ground slurry into a 1.5ml centrifuge tube and let it stand at room temperature for 5 minutes.

[0088...

Embodiment 2

[0115] Example 2. Synthesis of α-type Cono polypeptide

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

[0117] Table 3 Amino acid sequence of α type Cono 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 bond connection mode in each polypeptide is C1-C3, C2-C4. * Indicates amidation (CONH 2 ).

[0120] 1. Peptides

[0121] (1) Synthesis of T-1 (artificial synthesis using Fmoc method)

[0122] 1. Synthesis of peptide resin

[0123] Use solid phase synthesis technology. It was carried ...

Embodiment 3

[0172] Example 3 Determination of analgesic activity of α-type Cono polypeptide

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

[0174] α-Cono polypeptide Vc1.1: GCSDPRCNYDHPEICONH2.

[0175] 1. Synthesis of peptide resin

[0176] Use solid phase synthesis technology. It was carried out on the 433A peptide synthesizer (ABI American Applied System Biology Inc.). The amino acid used in the synthesis is Fmoc protected amino acid (product of Advanced Chemtech, USA), the resin used is Rink resin (product of Advanced Chemtech, USA), and the reagents used include DCC (Acros), HOBt (product of Advanced Chemtech, USA), NMP (PE company), piperidine (Shanghai Jier 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 (-Trt), and the side chain protecting groups of Arg are 9-phenylfluorenylmethyl (-Pbf) and the...

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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 a patent application with an application number of 201010121879.7, an application date of March 10, 2010, and an invention title of "α-type Cono polypeptide and its application". Technical field [0002] The invention relates to an α-type cono polypeptide and its application. Background technique [0003] Cono is a gastropod mollusk. There are more than 500 species in the world, all over the warm sea areas of the world. There are more than 100 types of Cono, mainly distributed in my country's Paracel Islands, Hainan Island and Taiwan waters. Cono polypeptides are secreted by the venom tube of the venom tube and the venom glands in the inner wall of the venom sac. The venom of each type of Cono contains 50-200 active peptides. Due to different sea areas, the toxin composition of snails may also vary. It is theoretically estimated that there are more than 50,000 peptides with different activities (McIntosh, JM, Jones, RM (2001)....

Claims

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Application Information

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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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