CD123-binding polypeptides and uses thereof
Provided herein are VHH-containing polypeptides that bind CD123. Uses of the VHH-containing polypeptides are also provided.
1. An immunoconjugate comprising a polypeptide comprising at least one VHH domain that binds CD123 and a cytotoxic agent, wherein at least one VHH domain that binds CD123 comprises:
a) a CDR1 comprising the amino acid sequence of SEQ ID NO: 3; a CDR2 comprising the amino acid sequence of SEQ ID NO: 4; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 5, and wherein the VHH domain comprises an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 26;
b) a CDR1 comprising the amino acid sequence of SEQ ID NO: 42; a CDR2 comprising the amino acid sequence of SEQ ID NO: 43; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 44, and wherein the VHH domain comprises an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 29 or 30;
c) 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: 9or 38, and wherein the VHH domain comprises an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 27;
d) a CDR1 comprising the amino acid sequence of SEQ ID NO: 15; a CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 17, and wherein the VHH domain comprises an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 28; or
e) a CDR1 comprising the amino acid sequence of SEQ ID NO: 23; a CDR2 comprising the amino acid sequence of SEQ ID NO: 24or 94; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 25, and wherein the VHH domain comprises an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 31, 32 or 92.
2. The immunoconjugate of claim 1 , wherein at least one VHH domain is humanized.
3. The immunoconjugate of claim 1 , wherein at least one VHH domain comprises an amino acid sequence at least 90%, at least 95%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 32, 29, 26, 27, 28, 30, 31, or 92.
4. The immunoconjugate of claim 1 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 32, 29, 26, 27, 28, 30, 31, or 92.
5. The immunoconjugate of claim 1 , comprising one, two, or three VHH domains.
6. The immunoconjugate of claim 5 , wherein each VHH domain binds CD123.
7. The immunoconjugate of claim 1 , comprising one VHH domain.
8. The immunoconjugate of claim 1 , wherein the polypeptide comprises an Fc region.
9. The immunoconjugate of claim 8 , wherein the Fc region comprises an amino acid sequence selected from SEQ ID NOs: 54-89.
10. The immunoconjugate of claim 8 , wherein the polypeptide forms a dimer under physiological conditions.
11. The immunoconjugate of claim 1 , wherein the CD123 is human CD123 comprising the amino acid sequence of SEQ ID NO: 1.
12. The immunoconjugate of claim 1 , 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.
13. A pharmaceutical composition comprising the immunoconjugate of claim 1 , and a pharmaceutically acceptable carrier.
14. A method of treating cancer comprising administering to a subject with cancer a pharmaceutically effective amount of the immunoconjugate of claim 1 .
15. The method of claim 14 , 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.
16. The method of claim 14 , further comprising administering an additional therapy or therapeutic agent.
17. The method of claim 16 , 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.
18. An isolated T cell comprising a polypeptide comprising at least one VHH domain that binds CD123, wherein at least one VHH domain comprises a CDR1, a CDR2, and a CDR3, respectively comprising the amino acid sequences of SEQ ID NOs: 42, 43, and 44; 3, 4, and 5; 7, 8, and 9; 7, 8, and 38; 11, 12, and 13; 15, 16, and 17; 19, 20, and 21; 23, 24, and 25; or 23, 94, and 25.
19. The isolated T cell of claim 18 , wherein the polypeptide comprising at least one VHH domain that binds CD123 is a chimeric antigen receptor (CAR).
20. The isolated T cell of claim 19 , wherein the T cell is engineered to express the CAR.
21. The isolated T cell of claim 18 , wherein the T cell is a CD4+ or CD8+ T cell.
22. The isolated T cell of claim 18 , wherein the polypeptide comprises one, two, or three VHH domains.
23. A pharmaceutical composition comprising the isolated T cell of claim 18 , and a pharmaceutically acceptable carrier.
24. A method of treating cancer comprising administering to a subject with cancer a pharmaceutically effective amount of the isolated T cell of claim 18 .
25. The method of claim 24 , wherein the isolated T cell is autologous to the subject.
26. The method of claim 24 , 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.
27. The method of claim 24 , further comprising administering an additional therapy or therapeutic agent.
28. The method of claim 27 , 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, and an oncolytic virus.