Multivalent and multispecific OX40-binding fusion proteins
This invention relates generally to molecules that specifically engage OX40, a member of the TNF receptor superfamily (TNFRSF). More specifically this invention relates to multivalent and multispecific molecules that bind at least OX40.
1. An isolated polypeptide that binds human OX40 and comprises at least one VHH that binds to OX40, wherein the at least one VHH that binds to OX40 comprises a complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO: 36, a complementarity determining region 2 (CDR2) comprising the amino acid sequence of SEQ ID NO: 37, and a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 42.
2. The isolated polypeptide of claim 1 , wherein the isolated polypeptide is monospecific.
3. The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises at least two VHHs, and wherein at least one VHH of the isolated polypeptide binds DR5, GITR, or CD137.
4. The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises at least two VHHs, and wherein at least one VHH of the isolated polypeptide binds PDL1.
5. The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises at least two VHHs, and wherein at least two VHHs of the isolated polypeptide are operably linked via a linker polypeptide.
6. The isolated polypeptide of claim 5 , wherein the linker polypeptide consists of 5-20 amino acids.
7. The isolated polypeptide of claim 6 , wherein the linker polypeptide is composed predominantly of glycine and serine.
8. The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises an immunoglobulin Fc region polypeptide.
9. The isolated polypeptide of claim 8 , wherein the immunoglobulin Fc region polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-6.
10. The isolated polypeptide of claim 1 , wherein the at least one VHH that binds to OX40 is a humanized VHH.
11. The isolated polypeptide of claim 1 , wherein the at least one VHH that binds to OX40 comprises an amino acid sequence selected from SEQ ID NOs: 25 and 377-385.
12. The isolated polypeptide of claim 1 , wherein the isolated polypeptide is tetravalent.
13. The isolated polypeptide of claim 12 , wherein the isolated polypeptide is dimeric and each molecule of the dimer comprises the structure: VHH-Linker-VHH-Linker-Hinge-Fc, wherein each VHH is a humanized VHH.
14. The isolated polypeptide of claim 1 , wherein the isolated polypeptide is hexavalent.
15. The isolated polypeptide of claim 14 , wherein the isolated polypeptide is dimeric and each molecule of the dimer comprises the structure: VHH-Linker-VHH-Linker-VHH-Linker-Hinge-Fc, wherein each VHH is a humanized VHH.
16. The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 389-394.
17. An isolated nucleic acid that encodes the isolated polypeptide of claim 1 .
18. An isolated vector comprising the nucleic acid of claim 17 .
19. An isolated host cell comprising the nucleic acid of claim 17 .
20. An isolated host cell that expresses the polypeptide of claim 1 .