OX40-binding polypeptides and uses thereof
Provided herein are VHH-containing polypeptides that bind OX40. In some embodiments, VHH-containing polypeptides that bind and agonize OX40 are provided. Uses of the VHH-containing polypeptides are also provided.
1. A method of treating cancer comprising administering to a subject with cancer a pharmaceutically effective amount of a polypeptide comprising at least three VHH domains that bind OX40 and an Fc region, wherein each VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 12, and wherein the VHH domain comprises a FR2 comprising the amino acid sequence of SEQ ID NO: 22, wherein the cancer is selected from solid cancer, hematological cancer, or lymphatic cancer.
2. The method of claim 1 , wherein the cancer is selected from basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and central nervous system cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer; gastrointestinal cancer; glioblastoma; hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or renal cancer; larynx cancer; liver cancer; lung cancer; small-cell lung cancer; non-small cell lung cancer; adenocarcinoma of the lung; squamous carcinoma of the lung; melanoma; myeloma; neuroblastoma; oral cavity cancer; ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary gland carcinoma; sarcoma; skin cancer; squamous cell cancer; stomach cancer; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary system; vulval cancer; lymphoma; Hodgkin's lymphoma; non-Hodgkin's lymphoma; B-cell lymphoma; low grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; Waldenstrom's macroglobulinemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; and chronic myeloblastic leukemia.
3. The method of claim 1 , further comprising administering an additional therapeutic agent.
4. The method of claim 3 , wherein the additional therapeutic agent is an anti-cancer agent.
5. The method of claim 4 , wherein the anti-cancer agent is selected from a chemotherapeutic agent, an anti-cancer biologic, radiation therapy, CAR-T therapy, and an oncolytic virus.
6. The method of claim 3 , wherein the additional therapeutic agent is an anti-cancer biologic.
7. The method of claim 6 , wherein the anti-cancer biologic is an agent that inhibits PD-1 and/or PD-L1.
8. The method of claim 7 , wherein the anti-cancer biologic is selected from nivolumab, pidilizumab, pembrolizumab, durvalumab, atezolizumab, avelumab, AMP-224, BMS-936559, AMP-514, MDX-1105, TSR-042, STI-A1010, and STI-A1110.
9. The method of claim 7 wherein the agent that inhibits PD-1 and/or PDL1 is an antibody.
10. The method of claim 9 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 14 and an Fc domain.
11. The method of claim 9 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 15.
12. The method of claim 6 , wherein the anti-cancer biologic is an agent that inhibits VISTA, gpNMB, B7H3, B7H4, HHLA2, CD73, CTLA4, or TIGIT.
13. The method of claim 12 , wherein the anti-cancer agent is an antibody.
14. The method of claim 6 , wherein the anti-cancer biologic is a cytokine.
15. The method of claim 6 , wherein the anti-cancer agent is CAR-T therapy or an oncolytic virus.
16. The method of claim 1 , further comprising tumor resection and/or radiation therapy.
17. The method of claim 1 , wherein the cancer is a solid tumor.
18. The method of claim 1 , wherein the cancer is selected from non-small cell lung cancer, melanoma, head and neck cancer, gastric cancer, renal cell carcinoma, squamous cell cancer, and urothelial carcinoma.
19. The method of claim 18 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 14.
20. The method of claim 18 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 15.
21. The method of claim 1 , wherein each VHH domain comprises a FR3 comprising the amino acid sequence of SEQ ID NO: 23.
22. The method of claim 1 , wherein each VHH domain comprises the amino acid sequence of SEQ ID NO: 9.
23. The method of claim 1 , wherein the polypeptide is an agonist of human OX40 biological activity.
24. The method of claim 1 , wherein the Fc domain comprises an amino acid sequence selected from SEQ ID NOs: 25 and 26.
25. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 14.
26. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 15.
27. A method of:
(a) increasing CD4 + and/or CD8 + cell proliferation;
(b) increasing CD25 expression on CD4+ and/or CD8+ T cells;
(c) increasing CD71 expression on CD4 + and/or CD8 + cells;
(d) increasing NFκB signaling in CD4 + and/or CD8 + cells; and/or
(e) increasing IFNγ expression in CD4 + and/or CD8 + cells,
comprising contacting T cells with a polypeptide comprising at least one VHH domain that binds OX40, wherein the VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 12, and wherein the VHH domain comprises a FR2 comprising the amino acid sequence of SEQ ID NO: 22.
28. The method of claim 27 , wherein the CD4 + and/or CD8 + cells are in vitro.
29. The method of claim 27 , wherein the CD4 + and/or CD8 + cells are in vivo.
30. The method of claim 27 , wherein the CD4 + and/or CD8 + cells are in the presence of Treg cells.
31. The method of claim 27 , wherein the increase is at least 1.5-fold, at least 2-fold, at least 3-fold, or by at least 5-fold.
32. The method of claim 27 , wherein the VHH domain comprises a FR3 comprising the amino acid sequence of SEQ ID NO: 23.
33. The method of claim 27 , wherein the VHH domain comprises the amino acid sequence of SEQ ID NO: 9.
34. The method of claim 27 , wherein the polypeptide comprises two, three, or four VHH domains.
35. The method of claim 27 , wherein each VHH domain binds OX40.
36. The method of claim 35 , wherein each VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 12, and wherein each VHH domain comprises a FR2 comprising the amino acid sequence of SEQ ID NO: 22.
37. The method of claim 35 , wherein each VHH domain comprises the amino acid sequence of SEQ ID NO: 9.
38. The method of claim 27 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 14 and an Fc domain.
39. The method of claim 27 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 15.