Conotoxin peptide κ-CPTX-BTL04, preparation method therefor, and uses thereof

Patent metadata

Field	Value
Publication number	US-10179802-B2
Application number	US-201415539752-A
Country	US
Kind code	B2
Filing date	Dec 26, 2014
Priority date	Dec 26, 2014
Publication date	Jan 15, 2019
Grant date	Jan 15, 2019

Abstract

Provided are a conotoxin peptide κ-CPTx-btl04 and a derivative polypeptide. The amino acid sequence of the conotoxin peptide is indicated by SEQ ID NO: 1. Also provided are a conotoxin peptide preparation method and uses of the conotoxin peptide in the treatment of diseases related to a calcium ion channel.

First claim

The invention claimed is:

1. A purified conotoxin peptide κ-CPTx-btlo 4 , which is derived from a polypeptide having the amino acid sequence of SEQ ID NO:1 by substitution of one or more amino acids therein, wherein the substitution of the one or more amino acids is selected from the group consisting of:
(i) substitution of the serine at position 1 with threonine;
(ii) substitution of the leucine at position 2 with isoleucine or valine; and
(iii) substitution of the tryptophan at position 9 with tyrosine or phenylalanine.

2. A polynucleotide encoding the conotoxin peptide κ-CPTx-btlo 4 according to claim 1 .

3. A nucleic acid construct comprising the polynucleotide of claim 2 , and one or more control sequences operably linked thereto and being able to direct the production of the polypeptide in an expression host.

4. An expression vector comprising the nucleic acid construct of claim 3 .

5. A transformed cell into which the nucleic acid construct of claim 3 is transformed.

6. A method for inhibiting a calcium ion channel comprising administration of the conotoxin peptide κ-CPTx-btlo 4 according to claim 1 .

7. A method for producing the conotoxin peptide κ-CPTx-btlo 4 according to claim 1 , which comprises:
(1) synthesizing the linear peptide of the conotoxin peptide κ-CPTx-btlo 4 according to its amino acid sequence by solid-phase chemical synthesis;
(2) performing oxidative refolding of the linear peptide obtained in step (1) with glutathione method.

8. The method according to claim 7 , wherein the solid-phase chemical synthesis is fluorenylmethoxycarbonyl solid-phase chemical synthesis.

9. A transformed cell into which the expression vector of claim 4 is transformed.

10. The method according to claim 6 , wherein the calcium ion channel is a high-voltage activated calcium ion channel.

Assignees

• Bgi Shenzhen

Inventors

• Liu Jie
• Lin Zhilong
• Wen Bo
• Tong Ting
• Mo Fen
• Peng Chao
• Shi Qiong

Classifications

• C07K7/06 Primary

  having 5 to 11 amino acids · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• A61K38/08

  Peptides having 5 to 11 amino acids {(A61K38/043 - A61K38/046 take precedence)} · CPC title

• C07K1/04

  on carriers {(C07K1/003, C07K1/006 take precedence)} · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title