Antagonists targeting the TGF-β pathway
Antitumor antagonists that bind specifically to immune checkpoint regulators, angiogenesis pathway regulators and/or TGF pathway regulators are disclosed. Also disclosed are methods for treating proliferative disorders, infections, and immunological disorders with the antitumor antagonists described herein.
1. A protein comprising:
a first targeting domain comprising a mutated TGF-β1 RII extracellular domain (mutated ECD), wherein the mutated ECD comprises SEQ ID NO:89 with one or more substitutions and/or deletions that reduce proteolytic degradation or clipping; and
an immunoglobulin scaffold having an amino terminus and a carboxy terminus, and
wherein the mutated ECD comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 114-123 and 251-270.
2. The protein of claim 1 , wherein the first targeting domain is linked to the immunoglobulin scaffold via a peptide linker.
3. The protein of claim 1 , further comprising a second targeting domain that comprises an immune checkpoint regulator antagonist, a VEGF pathway antagonist, or a Tie2 tyrosine kinase receptor antagonist.
4. The protein of claim 3 , wherein the second targeting domain comprises an immune checkpoint regulator antagonist.
5. The protein claim 4 , wherein the second targeting domain specifically binds to PD-1.
6. The protein claim 4 , wherein the second targeting domain specifically binds to PD-L1.
7. The protein claim 4 , wherein the second targeting domain specifically binds to TIGIT.
8. The protein claim 4 , wherein the second targeting domain specifically binds to LAG-3.
9. The protein of claim 3 , wherein the second targeting domain comprises a VEGF pathway antagonist.
10. The protein of claim 9 , wherein the VEGF pathway antagonist specifically binds to VEGF-A.
11. The protein of claim 9 , wherein the VEGF pathway antagonist comprises VEGFR ECD.
12. The protein of claim 3 , wherein the second targeting domain comprises a Tie2 tyrosine kinase receptor antagonist.
13. The protein of claim 2 , wherein the second targeting domain comprises a SEQ ID NO: 157, 158, 167, 168 or 169.
14. The protein of claim 1 , wherein the immunoglobulin scaffold is an IgG1, IgG2 or IgG4 scaffold.
15. The protein of claim 14 , wherein the immunoglobulin scaffold containing an N297A mutation, a K447A mutation, or both.
16. The protein of claim 14 , wherein the immunoglobulin scaffold comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 100, 159-166 and 173-175.
17. The protein of claim 1 , wherein the mutated ECD comprises the amino acid sequence of SEQ ID NO: 263.
18. The protein of claim 1 , wherein the mutated ECD comprises the amino acid sequence of SEQ ID NO: 116.
19. A pharmaceutical composition comprising:
the protein of claim 1 ; and
a pharmaceutically acceptable carrier.
20. An expression vector comprising a nucleotide sequence encoding the protein of claim 1 .
21. A cell line that expressed the protein of claim 1 .
22. A method of treating a cell proliferative disorder in a subject, comprising:
administering to a subject in need thereof an effective amount of the protein of claim 1 .