Cyclic GMP-chelating peptides for subcellular targeting

Patent metadata

Field	Value
Publication number	US-11512294-B2
Application number	US-201816759703-A
Country	US
Kind code	B2
Filing date	Nov 2, 2018
Priority date	Nov 2, 2017
Publication date	Nov 29, 2022
Grant date	Nov 29, 2022

Abstract

The disclosure pertains to the field of molecular means capable of binding to, and preferably of chelating, cGMP, appropriate for use in vitro or in vivo and preferably capable of targeting specific cellular compartments. The polypeptides of the disclosure comprise a chimeric construction derived from the N terminus part of PKG-Iα and PKG-Iβ, and the two cGMP binding sites of the wild type PKG.

First claim

The invention claimed is:

1. A polypeptide comprising:
a chimeric peptide comprising at least a sequence corresponding to SEQ ID NO: 1 or a fragment thereof, and at least a sequence corresponding to SEQ ID NO: 2 or a fragment thereof;
a cGMP binding domain comprising the sequence as set forth in SEQ ID NO: 6; and,
wherein said polypeptide is devoid of any catalytic domain,
and its functional variants that bind cGMP and have a sequence with at least 80% identity with the sequence of said polypeptide and which derives from the sequence of said polypeptide by conservative substitutions.

2. The polypeptide of claim 1 , wherein the chimeric peptide comprises the sequence as set forth in SEQ ID NO: 3, 4 or 5.

3. The polypeptide of claim 1 , wherein the cGMP binding domain comprises the sequence as set forth in SEQ ID NO: 7 or the sequence as set forth in SEQ ID NO: 8.

4. The polypeptide of claim 1 , wherein the cGMP binding domain comprises the sequence as set forth in SEQ ID NO: 9 or the sequence as set forth in SEQ ID NO: 10.

5. The polypeptide of claim 1 , wherein the chimeric peptide and the cGMP binding domain form a contiguous sequence.

6. The polypeptide of claim 1 , wherein the polypeptide comprises a peptide signal.

7. The polypeptide of claim 6 , wherein the peptide signal which comprises tandem repeats of the sequence as set forth in SEQ ID NO: 7.

8. The polypeptide of claim 1 , wherein the polypeptide comprises a fluorescent peptide.

9. The polypeptide of claim 2 , wherein the chimeric peptide comprises the sequence as set forth in SEQ ID NO: 3.

10. The polypeptide of claim 5 , wherein a C-terminal end of the chimeric peptide is fused to a N-terminal end of the cGMP binding domain.

11. The polypeptide of claim 8 , wherein the fluorescent peptide is selected from the group consisting of green fluorescent protein (GFP), cyan fluorescent protein (CFP), yellow fluorescent protein (YFP), red fluorescent protein (RFP), and variants thereof.

Assignees

• Univ Sorbonne
• Centre Nat Rech Scient
• Inst Nat Sante Rech Med
• Sorbaoone Univ
• Centre National De La Recharche Scient
• Institut National De La Sante Et La Rech Medicale Inserm

Inventors

• Nicol Xavier
• Ros Torres Oriol

Classifications

• C12N9/12 Primary

  transferring phosphorus containing groups, e.g. kinases (2.7) · CPC title

• C07K14/00

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof · CPC title

• C07K2319/60

  containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP] · 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

• C12Y207/01037

  Protein kinase (2.7.1.37) (C12Y207/11001, C12Y207/11008 - C12Y207/11013, C12Y207/11021, C12Y207/11022, C12Y207/11024, C12Y207/11025, C12Y207/1103, C12Y207/12001 take precedence) · CPC title