IP Library › Granted Patent US 12680093
Granted Patent B2
US 12680093 · App. 17/615,633 · Granted Jul 14, 2026

Protease substrate, and polypeptide including protease cleavage sequence

Inventors: Mika Sakurai (Shizuoka, JP); Tatsuya Kawa (Shizuoka, JP); Hitoshi Katada (Shizuoka, JP); Naoka Hironiwa (Singapore, JP)
Assignee: CHUGAI SEIYAKU KABUSHIKI KAISHA
C12N9/6462C07K7/04C07K16/2866C12N15/79A61K2039/505C07K2317/569C07K2319/50C12N2800/107
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Quick Facts
Patent No.
US 12680093
App. No.
17/615,633
Granted
Jul 14, 2026
Kind
B2
Abstract

The present application relates to protease substrates, peptide sequences cleavable by a protease, polypeptides comprising a protease cleavage sequence and methods for production thereof, pharmaceutical compositions comprising a polypeptide comprising a protease cleavage sequence, and methods for releasing an antigen-binding domain or a ligand by the cleavage of a protease cleavage sequence included in a polypeptide.

Claims (22)

1 . A protease substrate comprising a sequence selected from the following:

the sequence spanning from the N-terminal fourth to fifteenth amino acids of a sequence selected from SEQ ID NOs: 430, 400, 8, 26, 107, 114, 117, 130, 133, 135, 143, 149, 155, 156, 170, 172, 179, 184, 191, 203, 225, 265, 268, 271, 339, 359, and 425;

the sequence spanning from the N-terminal fourth to thirteenth amino acids of a sequence selected from SEQ ID NOs: 430, 400, 8, 26, 107, 114, 117, 130, 133, 135, 143, 149, 155, 156, 170, 172, 179, 184, 191, 203, 225, 265, 268, 271, 339, 359, and 425; or

the sequence spanning from the N-terminal sixth to thirteenth amino acids of a sequence selected from SEQ ID NOs: 430, 400, 8, 26, 107, 114, 117, 130, 133, 135, 143, 149, 155, 156, 170, 172, 179, 184, 191, 203, 225, 265, 268, 271, 339, 359, and 425.

2 . A method for releasing a ligand bound to a ligand-binding molecule, wherein the method comprises cleaving, by protease, a portion contained in the ligand-binding molecule capable of binding to the ligand, wherein the portion has at least one sequence selected from the following:

the sequence spanning from the N-terminal fourth to fifteenth amino acids of a sequence selected from SEQ ID NOs: 430, 400, 8, 26, 107, 114, 117, 130, 133, 135, 143, 149, 155, 156, 170, 172, 179, 184, 191, 203, 225, 265, 268, 271, 339, 359, and 425;

the sequence spanning from the N-terminal fourth to thirteenth amino acids of a sequence selected from SEQ ID NOs: 430, 400, 8, 26, 107, 114, 117, 130, 133, 135, 143, 149, 155, 156, 170, 172, 179, 184, 191, 203, 225, 265, 268, 271, 339, 359, and 425;

the sequence spanning from the N-terminal sixth to thirteenth amino acids of a sequence selected from SEQ ID NOs: 430, 400, 8, 26, 107, 114, 117, 130, 133, 135, 143, 149, 155, 156, 170, 172, 179, 184, 191, 203, 225, 265, 268, 271, 339, 359, and 425.

3 . A polypeptide comprising the protease substrate of claim 1 , wherein the polypeptide comprises an antigen-binding domain and a carrying moiety, and wherein the carrying moiety has an inhibiting domain for inhibiting the antigen-binding activity of the antigen-binding domain.

4 . A polypeptide comprising the protease substrate of claim 1 , wherein the polypeptide comprising the protease cleavage sequence is a ligand-binding molecule capable of binding to a ligand, and wherein the binding of the ligand-binding molecule to the ligand in a state where the protease cleavage sequence is cleaved is attenuated compared to the binding of the ligand-binding molecule to the ligand in a state where the protease cleavage sequence is not cleaved.

5 . A polypeptide comprising the protease substrate of claim 1 , wherein the polypeptide is a ligand-binding molecule capable of binding to a ligand, wherein the ligand-binding molecule comprises a single-domain antibody, wherein the single-domain antibody is capable of binding to a ligand and is introduced with at least one of the protease cleavage sequences, wherein the binding of the ligand-binding molecule to the ligand in a state where the protease cleavage sequence is cleaved is attenuated compared to the binding of the ligand-binding molecule to the ligand in a state where the protease cleavage sequence is not cleaved.

6 . The polypeptide of claim 4 which is a fusion protein, wherein the polypeptide is fused to the ligand.

7 . A pharmaceutical composition comprising the polypeptide of claim 3 .

8 . A pharmaceutical composition comprising the polypeptide of claim 4 and the ligand.

9 . A polynucleotide encoding the polypeptide of claim 3 .

10 . A vector comprising the polynucleotide according to claim 9 .

11 . A host cell comprising the polynucleotide according to claim 9 .

12 . A method for producing a polypeptide comprising culturing the host cell according to claim 11 .

13 . A polynucleotide encoding the protease substrate of claim 1 .

14 . A vector comprising the polynucleotide according to claim 13 .

15 . A host cell comprising the polynucleotide according to claim 13 .

16 . A method for producing a protease substrate comprising culturing the host cell according to claim 15 .