Granted Patent B2

US 12,617,829 · App. 18/601,474 · Granted May 5, 2026

Inventors: William Winston (West Newton, MA); Heather Brodkin (West Newton, MA); Cynthia Seidel-Dugan (Belmont, MA); Daniel Hicklin (Boston, MA); Jose Andres Salmeron-Garcia (Acton, MA)

Assignee: Werewolf Therapeutics, Inc.

C07K14/555C07K14/7156C07K14/5434C07K14/55C07K16/2818C07K16/2827C07K2319/31

Abstract

Claims (21)

1 . A conditionally active IL-18 comprising a fusion polypeptide comprising

at least one of each of:

a) A human IL-18 polypeptide [A];

b) an IL-18 blocking moiety [D], wherein the IL-18 blocking moiety [D] comprises an antibody or antigen-binding fragment that binds the IL-18 polypeptide, a ligand-binding domain or fragment of a cognate receptor for the IL-18 polypeptide;

c) a half-life extension element [H], wherein the half-life extension element [H] is a human serum albumin, an antigen-binding polypeptide that binds serum albumin, or an immunoglobulin Fc; and

d) a protease cleavable polypeptide linker [L];

wherein the conditionally active cytokine has attenuated IL-18 receptor activating activity that is at least about 10-fold less than the IL-18 receptor activating activity of the polypeptide that contains the IL-18 polypeptide that is produced by cleavage of the protease cleavable linker.

2 . The conditionally active cytokine of claim 1 , wherein the IL-18 blocking moiety [D] is an antibody fragment that binds to the IL-18 polypeptide and the antibody fragment is a single domain antibody, a Fab or scFv that binds the IL-18 polypeptide.

3 . The conditionally active cytokine of claim 1 , wherein the IL-18 blocking moiety inhibits the cytokine polypeptide from activating its cognate receptor.

4 . The conditionally active cytokine comprising of claim 1 , wherein the protease cleavable linker comprises a sequence that is capable of being cleaved by a protease selected from the group consisting of a kallikrein, thrombin, chymase, carboxypeptidase A, an elastase, PR-3, granzyme M, a calpain, a matrix metalloproteinase (MMP), an ADAM, a FAP, a plasminogen activator, a cathepsin, a caspase, a tryptase, and a tumor cell surface protease.

5 . The conditionally active cytokine of claim 1 , wherein the protease cleavable polypeptide linker independently comprises two or more cleavage sites for the same protease, or two or more cleavage sites that are cleaved by different proteases or at least one of the protease-cleavable polypeptides comprises a cleavage site for two or more different proteases.

6 . The conditionally active cytokine of claim 1 , wherein the protease cleavable polypeptide linker comprises a sequence that is cleaved by cathepsin selected from the group consisting of cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, and cathepsin G.

7 . The conditionally active cytokine of claim 1 , wherein the protease cleavable polypeptide linker comprises a sequence that is cleaved by a matrix metalloprotease (MMP) selected from the group consisting of MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, and MMP14.

8 . The conditionally active cytokine of claim 1 , comprising a fusion polypeptide having the Formula:

[A]-[L1]-[H]-[L2]-[D] or [A]-[L1]-[D]-[L2]-[H] or [D]-[L2]-[H]-[L1]-[A] or [H]-[L2]-[D]-[L1]-[A] or [D]-[L1]-[A]-[L1]-[H] or [H]-[L1]-[A]-[L1]-[D],

wherein, L1 is a protease cleavable polypeptide linker, and L2 is a polypeptide linker that is optionally protease cleavable.

9 . The conditionally active cytokine of claim 1 , wherein the serum half-life of the IL-18 polypeptide that is produced by cleavage of the protease-cleavable linker is comparable to the half-life of naturally occurring IL-18.

10 . A nucleic acid encoding the conditionally active cytokine of claim 1 .

11 . A pharmaceutical composition comprising a non-viral delivery system comprising the nucleic acid of claim 10 .

12 . A pharmaceutical composition comprising a mammalian cell comprising the conditionally active cytokine of claim 1 .

13 . A pharmaceutical composition comprising a mammalian cell comprising the nucleic acid of claim 10 .

Assignments (1)

Continuity (9)

References Cited (379)