Activatable interleukin-2 polypeptides and methods of use thereof
The disclosure features fusion proteins that are conditionally active variants of IL-2. In one aspect, the full-length polypeptides of the invention have reduced or minimal cytokine-receptor activating activity even though they contain a functional cytokine polypeptide. Upon activation, e.g., by cleavage of a linker that joins a blocking moiety, e.g., a steric blocking polypeptide, in sequence to the active cytokine, the cytokine can bind its receptor and effect signaling.
1. A pharmaceutical composition comprising at least one of each of:
a) a human interleukin 2 (IL-2) polypeptide [A];
b) a half-life extension element [B], wherein the half-life extension element is human serum albumin, an antigen-binding polypeptide that binds human serum albumin, or an immunoglobulin Fc;
c) an IL-2 blocking moiety [D], wherein the IL-2 blocking moiety comprises a ligand-binding domain or fragment of a cognate receptor for the IL-2 polypeptide, an antibody or antigen-binding fragment of an antibody that binds the IL-2 polypeptide; and
d) a protease-cleavable polypeptide linker [L];
wherein the IL-2 polypeptide and the IL-2 blocking moiety are operably linked by the protease-cleavable polypeptide linker and the IL-2 polypeptide has attenuated IL-2-receptor activating activity at least about 10 fold less than the IL-2-receptor activating activity of the polypeptide that comprises the IL-2 polypeptide that is produced by cleavage of the protease-cleavable polypeptide linker, and wherein the IL-2 receptor activating activity is assessed using a CTLL-2 proliferation assay, a phospho STAT ELISA, or HEK Blue reporter cell assay and using equal amounts, on a mole basis, of the IL-2 polypeptide and the pharmaceutical composition.
2. The pharmaceutical composition of claim 1 , wherein the serum half-life of the IL-2 polypeptide that is produced by cleavage of the protease-cleavable polypeptide linker is comparable to the half-life of naturally occurring IL-2.
3. The pharmaceutical composition of claim 1 , wherein the IL-2 polypeptide comprises an IL-2 variant or mutein.
4. The pharmaceutical composition of claim 1 , further comprising a tumor-specific antigen binding peptide, wherein the tumor-specific antigen binding peptide is linked to any one of [A], [B], or [D] by a cleavable linker.
5. The pharmaceutical composition of claim 4 , wherein the tumor-specific antigen binding peptide is linked to the IL-2 polypeptide by a non-cleavable linker and wherein the IL-2 polypeptide is linked to the half-life extension element or the IL-2 blocking moiety by a cleavable linker.
6. The pharmaceutical composition of claim 1 , comprising a plurality of protease-cleavable polypeptide linkers, wherein each protease-cleavable polypeptide linker independently comprises at least one sequence that is capable of being cleaved by a protease selected from the group consisting of a kallikrein, thrombin, chymase, carboxypeptidase A, cathepsin G, cathepsin L, an elastase, PR-3, granzyme M, a calpain, a matrix metalloproteinase (MMP), a fibroblast activation protein (FAP), an ADAM metalloproteinase, a plasminogen activator, a cathepsin, a caspase, a tryptase, and a tumor cell surface protease.
7. The pharmaceutical composition of claim 1 , wherein IL-2 blocking moiety inhibits activation of the IL-2 receptor by the human IL-2 polypeptide.
8. The pharmaceutical composition of claim 1 , wherein the antibody fragment that binds to the IL-2 polypeptide is a single domain antibody, Fab or scFv.
9. The pharmaceutical composition of claim 1 , wherein the IL-2 polypeptide comprises a naturally-occurring IL-2 polypeptide sequence or a functional fragment thereof.
10. The pharmaceutical composition 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.
11. The pharmaceutical composition 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.
12. The pharmaceutical composition of claim 1 , wherein the IL-2 polypeptide that is produced by cleavage of the protease-cleavable polypeptide linker does not comprise the half-life extension element or IL-2 blocking moiety.
13. The pharmaceutical composition of claim 1 , wherein the cognate receptor for the IL-2 is a alpha chain of the IL-2 receptor (IL-2Rα), beta chain of the IL-2 receptor (IL-2Rβ) or the gamma chain of the IL-2 receptor (IL-2Rγ).