Multivalent and multispecific DR5-binding fusion proteins and methods of treating neoplasms
The disclosure relates generally to molecules that specifically engage death receptor 5 (DR5), a member of the TNF receptor superfamily (TNFRSF) and methods of treatment. More specifically the disclosure relates to multivalent and multispecific molecules that bind at least DR5.
1. A method of treating a neoplasm that expresses death receptor 5 (DR5) comprising administering to a subject in need thereof a therapeutically effective amount of an isolated polypeptide that binds DR5 and comprises a plurality of DR5 binding domains (DR5BDs), wherein each DR5BD is a VHH comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 128, a CDR2 comprising the amino acid sequence of SEQ ID NO: 131, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 130, and wherein adjacent DR5BDs are operably linked by an amino acid linker.
2. The method of claim 1 , wherein the plurality of DR5BDs is two DR5BDs.
3. The method of claim 1 , wherein the plurality of DR5BDs is four DR5BDs.
4. The method of claim 1 , wherein the plurality of DR5BDs is six DR5BDs.
5. The method of claim 1 , wherein each DR5BD comprises the amino acid sequence of SEQ ID NO: 87.
6. The method of claim 2 , wherein each DR5BD comprises the amino acid sequence of SEQ ID NO: 87.
7. The method of claim 3 , wherein each DR5BD comprises the amino acid sequence of SEQ ID NO: 87.
8. The method of claim 4 , wherein each DR5BD comprises the amino acid sequence of SEQ ID NO: 87.
9. The method of claim 3 , wherein the polypeptide is a homodimer of the structure: DR5BD-Linker-DR5BD-Linker-Hinge-Fc, where each DR5BD is a humanized VHH sequence.
10. The method of claim 9 , wherein each DR5BD comprises the amino acid sequence of SEQ ID NO: 87.
11. The method of claim 1 , wherein the isolated polypeptide comprises an immunoglobulin hinge region and an immunoglobulin Fc region.
12. The method of claim 11 , wherein the immunoglobulin Fc region is an IgG1 Fc region, an IgG2 Fc region, an IgG3 Fc region, or an IgG4 Fc region.
13. The method of claim 11 , wherein the immunoglobulin Fc region comprises an amino acid sequence selected from SEQ ID NOs: 1-5 or 127.
14. The method of claim 1 , wherein each VHH is a humanized VHH.
15. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 113.
16. The method of claim 15 , wherein the polypeptide is a homodimer of the amino acid sequence of SEQ ID NO: 113 fused to an Fc region polypeptide.
17. A method of treating a neoplasm that expresses death receptor 5 (DR5) comprising administering to a subject in need thereof a therapeutically effective amount of an isolated polypeptide that binds DR5, wherein the polypeptide is a homodimer of the amino acid sequence of SEQ ID NO: 113 fused to an Fc region polypeptide of SEQ ID NO: 2.
18. The method of claim 1 , wherein each amino acid linker consists of 5-20 amino acids.
19. The method of claim 18 , wherein each amino acid linker is composed predominantly of glycine and serine.
20. The method of claim 19 , wherein each amino acid linker comprises an amino acid sequence selected from GGSGGS (SEQ ID NO: 11); GGSGGSGGS (SEQ ID NO: 12); GGSGGSGGSGGS (SEQ ID NO: 13); and GGSGGSGGSGGSGGS (SEQ ID NO: 14).
21. The method of claim 11 , wherein the immunoglobulin hinge region comprises an amino acid sequence selected from EPKSSDKTHTCPPC (SEQ ID NO: 6), DKTHTCPPC (SEQ ID NO: 7), and ESKYGPPCPPC (SEQ ID NO: 8).
22. The method of claim 10 , wherein each amino acid linker consists of 5-20 amino acids.
23. The method of claim 22 , wherein each amino acid linker is composed predominantly of glycine and serine.
24. The method of claim 23 , wherein each amino acid linker comprises an amino acid sequence selected from GGSGGS (SEQ ID NO: 11); GGSGGSGGS (SEQ ID NO: 12); GGSGGSGGSGGS (SEQ ID NO: 13); and GGSGGSGGSGGSGGS (SEQ ID NO: 14).
25. The method of claim 1 , wherein the neoplasm is cancer.
26. The method of claim 3 , wherein the neoplasm is cancer.
27. The method of claim 9 , wherein the neoplasm is cancer.
28. The method of claim 15 , wherein the neoplasm is cancer.
29. The method of claim 17 , wherein the neoplasm is cancer.
30. The method of claim 25 , wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.
31. The method of claim 25 , wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.
32. The method of claim 26 , wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.
33. The method of claim 26 , wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.
34. The method of claim 27 , wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.
35. The method of claim 27 , wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.
36. The method of claim 28 , wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.
37. The method of claim 28 , wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.
38. The method of claim 29 , wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.
39. The method of claim 29 , wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.