Conotoxin peptide k-cptx-btl05, preparation method therefor, and uses thereof

Patent metadata

Field	Value
Publication number	US-2018201650-A1
Application number	US-201415539763-A
Country	US
Kind code	A1
Filing date	Dec 26, 2014
Priority date	Dec 26, 2014
Publication date	Jul 19, 2018
Grant date	—

Abstract

Provided are a conotoxin peptide κ-CPTx-btl05 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

1 . A conotoxin peptide κ-CPTx-btl05, which is:
(a) a polypeptide having the amino acid sequence shown in SEQ ID NO: 1; or
(b) a polypeptide derived from polypeptide (a) by substitution, addition and deletion of one or more amino acids in the amino acid sequence of SEQ ID NO: 1 of polypeptide (a) and having the function of polypeptide (a); wherein the amino acids to be substituted, added or deleted do not include cysteine;
the amino acid sequences of polypeptides (a) and (b) contain two pairs of disulfide bonds.

2 . The conotoxin peptide κ-CPTx-btl05 according to claim 1 , wherein the substitution of the one or more amino acids is selected from the group consisting of:
(i) substitution of the isoleucine at position 2 with leucine or valine;
(ii) substitution of the valine at position 5 with leucine or isoleucine;
(iii) substitution of the threonine at position 8 or 10 with serine;
(iv) substitution of the leucine at position 16 with isoleucine or valine.

3 . A polynucleotide encoding the conotoxin peptide κ-CPTx-btl05 according to claim 1 .

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

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

6 . A transformed cell into which the nucleic acid construct of claim 4 .

7 . A method for inhibiting a calcium ion channel comprising administration of the conotoxin peptide κ-CPTx-btl05 according to claim 1 .

8 . (canceled)

9 . (canceled)

10 . A method for producing the conotoxin peptide κ-CPTx-btl05 according to claim 1 , which comprises:
(1) synthesizing the linear peptide of the conotoxin peptide κ-CPTx-btl05 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.

11 . The method according to claim 10 , wherein the solid-phase chemical synthesis is fluorenylmethoxycarbonyl solid-phase chemical synthesis.

12 . A transformed cell into which the expression vector of claim 5 is transformed.

13 . The method according to claim 7 , 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

• 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

• C07K7/08 Primary

  having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) · CPC title

• C07K1/04

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

• C07K1/06

  using protecting groups or activating agents {(C07K1/003, C07K1/006 take precedence)} · CPC title

• A61K38/10

  Peptides having 12 to 20 amino acids {(A61K38/043 - A61K38/046 take precedence)} · CPC title