Rationally designed protein compositions
Novel compositions and methods relating to or derived from a rationally designed fusion protein composition combines a therapeutic antibody with an IL2 mutant that can simultaneously enhance anti-tumor immunity or derepress tumor-associated immunosuppression along with direct activation of effector cells by IL2 without activating T reg are provided. The fusion protein can be used to prevent or therapeutically treat cancer.
1. A fusion protein, comprising a therapeutic protein connected to a mutant Interleukin-2 (IL2) polypeptide, wherein the therapeutic protein is an entire anti-PDL1 antibody, and wherein the mutant IL2 polypeptide comprises an amino acid sequence identical to SEQ ID NO: 25 or SEQ ID NO: 27.
2. The fusion protein of claim 1 , wherein the entire anti-PDL1 antibody comprises a pair of heavy chains and a pair of light chains, and wherein the fusion protein comprises two mutant IL2 polypeptides, each mutant IL2 polypeptide connected to one of the C-terminals of the heavy chains.
3. The fusion protein of claim 2 , wherein each mutant IL2 polypeptide is connected to the corresponding C-terminal of the heavy chains through a linker sequence.
4. The fusion protein of claim 3 , wherein the linker sequence is a serine-glycine linker.
5. The fusion protein of claim 4 , wherein the linker sequence is SEQ ID NO: 19.
6. The fusion protein of claim 1 , wherein the anti-PDL1 antibody has one or more of the following features: (a) it binds PDL1 with Kd<100 nM to both purified protein as well as protein expressed on cell surface; (b) it blocks PDL1-PD1 interaction in vitro; and/or (c) it de-represses PDL1-mediated repression of effector T cells in activated PBMC cultures.
7. The fusion protein of claim 1 , wherein the anti-PDL1 antibody is an antibody of any available isotype or any mutant forms of Fc region that mediates or enhances ADCC and/or CDC function.
8. The fusion protein of claim 1 , wherein the anti-PDL1 antibody is an antibody of IgG1 isotype.
9. The fusion protein of claim 1 , wherein the anti-PDL1 antibody comprises a heavy chain variable region and a light chain variable region where their respective sequences consist of pairings selected from the group consisting of: (a) SEQ ID NO: 3 and SEQ ID NO: 6; (b) SEQ ID NO: 9 and SEQ ID NO: 12; and (c) SEQ ID NO: 15 and SEQ ID NO: 18.
10. The fusion protein of claim 9 , wherein the anti-PDL1 antibody comprises a heavy chain and a light chain where their respective sequences are at least 95% identical to the pairings selected from the group consisting of: (a) SEQ ID NO: 2 and SEQ ID NO: 5; (b) SEQ ID NO: 8 and SEQ ID NO: 11; and (c) SEQ ID NO: 14 and SEQ ID NO: 17.
11. A pharmaceutical composition comprising the fusion protein of claim 1 , and a pharmaceutically acceptable excipient, carrier or diluent.
12. A nucleic acid molecule that encodes the fusion protein of claim 1 , wherein the nucleic acid molecule is a DNA molecule or RNA molecule.
13. A mammalian expression system that produces the fusion protein of claim 1 .
14. A mutant IL2 polypeptide that is less selective towards a high-affinity receptor of a wild type IL2, wherein the mutant IL2 polypeptide comprises an amino acid sequence identical to a sequence selected from the group consisting of SEQ ID NO: 25, and SEQ ID NO: 27.