CD33-binding polypeptides and uses thereof
Provided herein are VHH-containing polypeptides that bind CD33. Uses of the VHH-containing polypeptides are also provided.
1. A polypeptide comprising at least one VHH domain that binds CD33, wherein at least one VHH domain that binds CD33 comprises:
a) a CDR1 comprising the amino acid sequence of SEQ ID NO: 47; a CDR2 comprising the amino acid sequence of SEQ ID NO: 48; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 49;
b) a CDR1 comprising the amino acid sequence of SEQ ID NO: 7; a CDR2 comprising the amino acid sequence of SEQ ID NO: 8; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 9;
c) a CDR1 comprising the amino acid sequence of SEQ ID NO: 50; a CDR2 comprising the amino acid sequence of SEQ ID NO: 51; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 52;
d) a CDR1 comprising the amino acid sequence of SEQ ID NO: 59; a CDR2 comprising the amino acid sequence of SEQ ID NO: 60; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 61;
e) a CDR1 comprising the amino acid sequence of SEQ ID NO: 65; a CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 67;
f) a CDR1 comprising the amino acid sequence of SEQ ID NO: 68; a CDR2 comprising the amino acid sequence of SEQ ID NO: 69; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 70; or
g) a CDR1 comprising the amino acid sequence of SEQ ID NO: 71; a CDR2 comprising the amino acid sequence of SEQ ID NO: 72; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 73.
2. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 47; a CDR2 comprising the amino acid sequence of SEQ ID NO: 48; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 49.
3. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 7; a CDR2 comprising the amino acid sequence of SEQ ID NO: 8; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 9.
4. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 50; a CDR2 comprising the amino acid sequence of SEQ ID NO: 51; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 52.
5. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 59; a CDR2 comprising the amino acid sequence of SEQ ID NO: 60; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 61.
6. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 65; a CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 67.
7. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 68; a CDR2 comprising the amino acid sequence of SEQ ID NO: 69; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 70.
8. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 71; a CDR2 comprising the amino acid sequence of SEQ ID NO: 72; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 73.
9. The polypeptide of claim 1 , wherein at least one VHH domain is humanized.
10. The polypeptide of claim 1 , wherein at least one VHH domain comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 38, 114, 39, 42, 44, 45, 46, 115, 118, 120, 121, or 122.
11. The polypeptide of claim 1 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38, 114, 39, 42, 44, 45, 46, 115, 118, 120, 121, or 122.
12. The polypeptide of claim 1 , wherein at least one VHH domain comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 2, 6, 10, 18, 26, 30, 34, 123, 124, 126, 128, 129, 130, 131.
13. The polypeptide of claim 1 , comprising one, two, or three VHH domains.
14. The polypeptide of claim 1 , wherein the polypeptide comprises at least one binding domain that binds an antigen other than CD33.
15. The polypeptide of claim 14 , wherein the polypeptide comprises at least one binding domain that binds CD3, T-cell receptor (TCR) α, TCRβ, CD28, CD16, CD32A, CD64, CD89, NKp46, or NKG2D.
16. The polypeptide of claim 13 , wherein each VHH domain binds CD33.
17. The polypeptide of claim 1 , wherein the polypeptide comprises an Fc region.
18. The polypeptide of claim 17 , wherein the Fc region comprises an amino acid sequence selected from SEQ ID NOs: 74-109.
19. The polypeptide of claim 17 , which forms a dimer under physiological conditions.
20. An immunoconjugate comprising the polypeptide of claim 1 and a cytotoxic agent.
21. The immunoconjugate of claim 20 , wherein the cytotoxic agent is selected from a calicheamicin, an auristatin, a dolastatin, a tubulicin, a maytansinoid, a cryptophycin, a duocarmycin, an esperamicin, a pyrrolobenzodiazepine, and an enediyne antibiotic.
22. A pharmaceutical composition comprising the polypeptide of claim 1 , and a pharmaceutically acceptable carrier.
23. A method of treating cancer comprising administering to a subject with cancer a pharmaceutically effective amount of the polypeptide of claim 1 .
24. The method of claim 23 , wherein the cancer is selected from lymphoma; Hodgkin's lymphoma; non-Hodgkin's lymphoma; B-cell lymphoma; low grade/follicular non-Hodgkin's lymphoma (NEIL); 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); acute myeloid leukemia (AML); Hairy cell leukemia; and chronic myeloblastic leukemia.
25. The method of claim 23 , further comprising administering an additional therapy or therapeutic agent.
26. The method of claim 25 , wherein the additional therapy or therapeutic agent is an anti-cancer therapy or agent, wherein the anti-cancer therapy or agent is selected from a chemotherapeutic agent, an anti-cancer biologic, radiation therapy, CAR-T therapy, and an oncolytic virus.
27. The polypeptide of claim 1 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
28. The polypeptide of claim 14 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
29. The polypeptide of claim 16 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
30. The method of claim 23 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 47; a CDR2 comprising the amino acid sequence of SEQ ID NO: 48; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 49.
31. The method of claim 23 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
32. A method of treating cancer comprising administering to a subject with cancer a pharmaceutically effective amount of the polypeptide of claim 14 .
33. The method of claim 32 , wherein the cancer is selected from 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); acute myeloid leukemia (AML); Hairy cell leukemia; and chronic myeloblastic leukemia.
34. A method of treating cancer comprising administering to a subject with cancer a pharmaceutically effective amount of the polypeptide of claim 15 .
35. The method of claim 34 , wherein the cancer is selected from 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); acute myeloid leukemia (AML); Hairy cell leukemia; and chronic myeloblastic leukemia.
36. A method of treating cancer comprising administering to a subject with cancer a pharmaceutically effective amount of the immunoconjugate of claim 20 .
37. The method of claim 36 , wherein the cancer is selected from 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); acute myeloid leukemia (AML); Hairy cell leukemia; and chronic myeloblastic leukemia.