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 OX40 and comprises a plurality of Tumor Necrosis Factor receptor superfamily (TNFRSF) binding domain (TBDs), wherein at least a first TBD (TBD1) binds OX40.
2 . An isolated polypeptide that binds PDL1 and comprises a plurality of polypeptide binding domains (BDs), wherein at least a first BD (BD1) binds PDL1.
3 . An isolated polypeptide that binds PDL1 and OX40 and comprises a plurality of polypeptide binding domains (BDs) and a plurality of Tumor Necrosis Factor receptor superfamily (TNFRSF) binding domain (TBDs), wherein at least a first binding domain (BD1) binds PLD1 and at least a first TNFRSF binding domain (TBD1) binds OX40.
4 . The isolated polypeptide of claim 1 , wherein the isolated polypeptide is monospecific.
5 . The isolated polypeptide of claim 1 , wherein the isolated polypeptide is multispecific.
6 . The isolated polypeptide of claim 1 , wherein the isolated polypeptide is bispecific.
7 . The isolated polypeptide of claim 1 , the polypeptide comprises at least a second TBD (TBD2) that binds a second TNFRSF member.
8 . The isolated polypeptide of claim 1 , wherein each of TBD in the plurality of TBDs binds OX40.
9 . The isolated polypeptide of claim 8 , wherein the plurality of TBDs binds the same epitope on OX40.
10 . The isolated polypeptide of claim 8 , wherein at least two TBDs in the plurality of TBDs bind a different epitope on OX40.
11 . The isolated polypeptide of claim 8 , wherein the plurality of TBDs comprises at least four TBDs.
12 . The isolated polypeptide of claim 8 , wherein the plurality of TBDs comprises at least six TBDs.
13 . The isolated polypeptide of claim 2 , wherein the polypeptide comprises at least a second BD (BD2) that binds a second antigen.
14 . The isolated polypeptide of claim 2 , wherein each BD in the plurality of BDs binds PDL1.
15 . The isolated polypeptide of claim 14 , wherein the plurality of BDs binds the same epitope on PDL1.
16 . The isolated polypeptide of claim 14 , wherein at least two BDs in the plurality of BDs bind a different epitope on PDL1.
17 . The isolated polypeptide of claim 13 , wherein the plurality of BDs comprises at least four BDs.
18 . The isolated polypeptide of claim 13 , wherein the plurality of BDs comprises at least six BDs.
19 . The isolated polypeptide of claim 1 , wherein each of the TBD in the plurality of TBDs or each of the BDs in the plurality of BDs are operably linked via a linker polypeptide.
20 . The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises at least one binding domain that binds a target, provided that the target is not a TNFRSF member.
21 . The isolated polypeptide of claim 1 , wherein at least one of the TBD in the plurality of TBDs binds a second TNFRSF member selected from the group consisting of DR5, GITR, and CD137.
22 . The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises a heterodimerization domain.
23 . The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises an immunoglobulin Fc region polypeptide.
24 . The isolated polypeptide of claim 23 , wherein the immunoglobulin Fc region polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-6.
25 . The isolated polypeptide of claim 1 , wherein at least one TBD in the plurality of TBDs or at least one BD in the plurality of BDs comprises an antibody or antigen-binding fragment thereof.
26 . The isolated polypeptide of claim 1 , wherein each TBD in the plurality of TBDs or at least one BD in the plurality of BDs comprises an antibody or antigen-binding fragment thereof.
27 . The isolated polypeptide of claim 25 , wherein the antibody or antigen-binding fragment thereof is a scFv, a Fab, a single domain antibody (sdAb), a V NAR , or a VHH.
28 . The isolated polypeptide of claim 25 , wherein the antibody or antigen-binding fragment is a sdAb.
29 . The isolated polypeptide of claim 28 , wherein the sdAb is a human or humanized sdAb.
30 . The isolated polypeptide of claim 28 , wherein the sdAb is VHH, V NAR , an engineered VH domain or an engineered VK domain.
31 . The isolated polypeptide of claim 30 , wherein the sdAb is generated from a cartilaginous fish heavy chain only antibody.
32 . The isolated polypeptide of claim 1 , wherein at least one the TNFRSF binding domains comprises a non-antibody scaffold protein.
33 . The isolated polypeptide of claim 32 , wherein the non-antibody scaffold protein is an ankyrin repeat protein, a darpin, an avimer, an anticalin/lipocalin, a centyrin, or a fynomer.
34 . The isolated polypeptide of claim 1 , wherein the polypeptide comprises an amino acid sequence that binds OX40 selected from the group consisting of SEQ ID NO: 16-29 and 377-386.
35 . The isolated polypeptide of claim 1 , wherein the polypeptide comprises an amino acid sequence that binds OX40 and comprises a complementarity determining region 1 (CDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 30, 36, and 44; a complementarity determining region 2 (CDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 31, 35, and 37; and a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 32-34, 48-43, and 45.
36 . The isolated polypeptide of claim 2 , wherein the polypeptide comprises an amino acid sequence that binds PDL1 selected from the group consisting of SEQ ID NO: 52-57.
37 . The isolated polypeptide of claim 2 , wherein the polypeptide comprises an amino acid sequence that binds PDL1 and comprises a complementarity determining region 1 (CDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 58, 61, and 64; a complementarity determining region 2 (CDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 59, 62, 65, and 69; and a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 60, 63, 66-68, and 70.
38 . The isolated polypeptide of claim 3 , wherein the polypeptide comprises a first amino acid sequence that binds OX40 selected from the group consisting of SEQ ID NO: 52-57, and a second amino acid sequence that binds PDL1 selected from the group consisting of SEQ ID NO: 52-57.
39 . The isolated polypeptide of claim 3 , wherein the polypeptide comprises an amino acid sequence that binds OX40 and comprises a complementarity determining region 1 (CDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 30, 36, and 44; a complementarity determining region 2 (CDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 31, 35, and 37; and a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 32-34, 48-43, and 45, and wherein the polypeptide comprises an amino acid sequence that binds PDL1 and comprises a CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 58, 61, and 64; a CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 59, 62, 65, and 69; and a CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 60, 63, 66-68, and 70.
40 . The isolated polypeptide of claim 1 , wherein the polypeptide is tetravalent.
41 . The isolated polypeptide of claim 40 , wherein the polypeptide comprises the structure: VHH-Linker-VHH-Linker-Hinge-Fc, where the VHH is a humanized or fully human VHH sequence.
42 . The isolated polypeptide of claim 1 , wherein the polypeptide is hexavalent.
43 . The isolated polypeptide of claim 42 , wherein the polypeptide comprises the structure: VHH-Linker-VHH-Linker-VHH-Linker-Hinge-Fc, where the VHH is a humanized or fully human VHH sequence.
44 . An isolated polypeptide that binds OX40 and PDL1, wherein the polypeptide comprises an amino acid sequence that is selected from the group consisting of SEQ ID NO: 387-394.
45 . Use of the polypeptide of claim 1 for treating neoplasms.
46 . Use of the polypeptide of claim 1 for modulating immune cells to enhance tumor destruction.
47 . Use of the polypeptide of claim 1 for treating an inflammatory disorder.