A kind of α-type conus polypeptide and its application

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

Inactive Publication Date: 2011-12-21
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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  • A kind of α-type conus polypeptide and its application
  • A kind of α-type conus polypeptide and its application
  • A kind of α-type conus polypeptide and its application

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

[0173] Example 3 Determination of analgesic activity of α-type Cono polypeptide

[0174] 1. Preparation of α-Cono polypeptide Vc1.1

[0175] α-Cono polypeptide Vc1.1: GCSDPRCNYDHPEICONH 2 .

[0176] 1. Synthesis of peptide resin

[0177] 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).

[0178] 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 t...

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Abstract

The invention discloses seven alpha type conotoxin peptides and applications thereof. The peptide I has an amino acid sequence shown as the sequence 7 in a sequence table, and the last amino acid residue is amidated; the peptide II has an amino acid sequence shown as the sequence 8 in the sequence table, and the last amino acid residue is amidated; the peptide III has an amino acid sequence shownas the sequence 9 in the sequence table, and the last amino acid residue is amidated; the peptide IV has an amino acid sequence shown as the sequence 10 in the sequence table, and the last amino acidresidue is amidated; the peptide V has an amino acid sequence shown as the sequence 11 in the sequence table, and the last amino acid residue is amidated; the peptide VI has an amino acid sequence shown as the sequence 12 in the sequence table, and the last amino acid residue is amidated; and the peptide VII has an amino acid sequence shown as the sequence 13 in the sequence table. Proved by determination of activity, the seven peptides shown as a general formula I have an analgesic effect, wherein alpha type conotoxin peptides T-1, T-2, T-3 and T-4 have remarkable analgesic activity, and thealpha type conotoxin peptides T5-T7 also have remarkable analgesic activity, and the application value on the aspect of analgesia is achieved.

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. The active peptides in different varieties of Cono are different, even if the same species Due to different sea areas, the toxin composition of snails may also vary. It is theoretically estimated that the...

Claims

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Application Information

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