Conjugate comprising an active MMP-9-binding peptide

Patent metadata

Field	Value
Publication number	US-12043658-B2
Application number	US-202218057480-A
Country	US
Kind code	B2
Filing date	Nov 21, 2022
Priority date	Jul 18, 2017
Publication date	Jul 23, 2024
Grant date	Jul 23, 2024

Abstract

The present invention provides a novel peptide that has an amino acid sequence represented by SEQ ID NO: 18, and binds to an active protease but does not bind to a pro-protease.

First claim

The invention claimed is:

1. A conjugate comprising a SPINK2 mutant peptide and a moiety bound thereto, wherein the SPINK2 mutant peptide binds to an active human MMP-9 but does not bind to a pro human MMP-9, and wherein the SPINK2 mutant peptide comprises the amino acid sequence as set forth in any one of SEQ ID NO: 2 to 17.

2. The conjugate according to claim 1 , wherein the SPINK2 mutant peptide binds to the catalytic domain of the active human MMP-9.

3. The conjugate according to claim 1 , wherein the SPINK2 mutant peptide inhibits the protease activity of the active human MMP-9.

4. The conjugate according to claim 3 , wherein the inhibition is specific to human MMP-9.

5. The conjugate according to claim 1 , wherein the SPINK2 mutant peptide further comprises one of (a) to (c):
(a) an additional 1 to 3 amino acid residues;
(b) the amino acid sequence as set forth in SEQ ID NO: 19; or
(c) the amino acid sequence as set forth in SEQ ID NO: 20; and
wherein any one of (a) to (c) is linked to the N-terminal amino acid of any one of the sequences as set forth in SEQ ID NO: 2 to 17.

6. The conjugate according to claim 1 , wherein the SPINK2 mutant peptide further comprises an additional 1 to 6 amino acid residues, wherein the additional 1 to 6 amino acid residues are linked to the C-terminal amino acid of any one of the sequences as set forth in SEQ ID NO: 2 to 17.

7. The conjugate according to claim 1 , wherein the SPINK2 mutant peptide further comprises one of (a) to (c):
(a) the amino acid sequence as set forth in SEQ ID NO: 21;
(b) a three amino acid sequence consisting of Gly-Gly-Gly; or
(c) a two amino acid sequence consisting of Gly-Gly; and
wherein any one of (a) to (c) is linked to the C-terminal amino acid of any one of the sequences as set forth in SEQ ID NO: 2 to 17.

8. The conjugate according to claim 1 , wherein the SPINK2 mutant peptide comprises three disulfide bonds, a loop structure, an α-helix, and a β-sheet.

9. The conjugate according to claim 1 , wherein the conjugate is a peptide.

10. The conjugate according to claim 1 , wherein the conjugate comprises an immunoglobulin Fc region.

11. The conjugate according to claim 1 , wherein the moiety comprises at least one drug.

12. The conjugate according to claim 1 , wherein the moiety comprises at least one labeling molecule.

13. The conjugate according to claim 12 , wherein the labeling molecule comprises an enzyme label, a radioactive label, a color label, a fluorescent label, a chromogenic label, a luminescent label, a hapten molecule, a biotin molecule, a metal complex label, a metal molecule, or a colloidal gold molecule.

14. A composition comprising the conjugate of claim 1 .

15. A pharmaceutical composition comprising the conjugate according to claim 1 .

16. A method for producing a SPINK2 mutant peptide that comprises the amino acid sequence as set forth in any one of SEQ ID NO: 2 to 17, wherein the method comprises:
(i) culturing a cell containing a polynucleotide having a nucleotide sequence encoding a SPINK2 mutant peptide that comprises the amino acid sequence as set forth in any one of SEQ ID NO: 2 to 17 or a vector into which the polynucleotide has been inserted; and
(ii) recovering the SPINK2 mutant peptide from the culture.

17. A method for producing the conjugate according to claim 1 , the method comprising a step of preparing the conjugate by chemical synthesis or in vitro translation.

18. The method according to claim 16 , wherein step (ii) comprises recovering the SPINK2 mutant peptide by affinity purification.

Assignees

• Daiichi Sankyo Co Ltd

Inventors

• YANO Hidenori
• Nishimiya Daisuke
• Hashimoto Ryuji
• Niitsu Yoichi

Classifications

• G01N33/68

  involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)} · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C07K14/001

  by chemical synthesis · 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

• A61K47/68

  the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment · CPC title