CLEC12a binding polypeptides and uses thereof
Provided herein are VHH-containing polypeptides that bind CLEC12a. Uses of the VHH-containing polypeptides are also provided.
1. A polypeptide comprising at least one VHH domain that binds CLEC12a, wherein each VHH domain comprises a CDR1, a FR2, a CDR2, a FR3, and a CDR3, in that order, and wherein at least one VHH domain that binds CLEC12a comprises:
a CDR1 comprising the amino acid sequence of SEQ ID NO: 32; a CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 34 or 5.
2. The polypeptide of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 32; a CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 34.
3. The polypeptide of claim 1 , wherein at least one VHH domain is humanized.
4. The polypeptide of claim 1 , wherein at least one VHH domain that binds CLEC12a comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 26 or 94.
5. The polypeptide of claim 1 , wherein at least one VHH domain that binds CLEC12a comprises the amino acid sequence of SEQ ID NO: 26 or 94.
6. The polypeptide of claim 1 , comprising one, two, or three VHH domains, wherein each VHH domain binds CLEC12a.
7. The polypeptide of claim 6 , wherein at least one VHH domain that binds CLEC12a comprises the amino acid sequence of SEQ ID NO: 26 or 94.
8. The polypeptide of claim 1 , wherein the polypeptide comprises at least one binding domain that binds an antigen other than CLEC12a, wherein the at least one binding domain binds CD3, T-cell receptor (TCR)α, TCRβ, CD28, CD16, CD32A, CD64, CD89, NKp46, or NKG2D.
9. The polypeptide of claim 8 , wherein at least one VHH domain that binds CLEC12a comprises the amino acid sequence of SEQ ID NO: 26 or 94.
10. The polypeptide of claim 1 , wherein the polypeptide comprises an Fc region.
11. The polypeptide of claim 10 , wherein the Fc region comprises an amino acid sequence selected from SEQ ID NOs: 50 to 85.
12. The polypeptide of claim 10 , which is a homodimer under physiological conditions.
13. An immunoconjugate comprising the polypeptide of claim 1 linked to a cytotoxic agent.
14. The immunoconjugate of claim 13 , 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.
15. The immunoconjugate of claim 13 , wherein at least one VHH domain that binds CLEC12a comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 26 or 94.
16. A method of inhibiting or slowing the progression of cancer comprising administering to a subject with cancer a pharmaceutically effective amount of the immunoconjugate of claim 15 , wherein the cancer is selected from chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); acute myeloid leukemia (AML); Hairy cell leukemia; and chronic myeloblastic leukemia.
17. The method of claim 16 , 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.
18. The method of claim 17 , wherein the polypeptide comprises at least one VHH domain that binds CLEC12a comprising the amino acid sequence of SEQ ID NO: 26 or 94.
19. The method of claim 16 , wherein the polypeptide comprises at least one VHH domain that binds CLEC12a comprising the amino acid sequence of SEQ ID NO: 26 or 94.
20. The immunoconjugate of claim 13 , wherein at least one VHH domain that binds CLEC12a comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 26 or 94.
21. A pharmaceutical composition comprising the polypeptide of claim 1 , and a pharmaceutically acceptable carrier.
22. The polypeptide of claim 1 , wherein at least one VHH domain that binds CLEC12a comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 26 or 94.
23. A method of inhibiting or slowing the progression of cancer comprising administering to a subject with cancer a pharmaceutically effective amount of a polypeptide comprising at least one VHH domain that binds CLEC12a, and wherein at least one VHH domain that binds CLEC12a comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 32; a CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 34 or 5, and is at least 95% identical to the amino acid sequence of SEQ ID NO: 26 or 94, wherein the polypeptide comprises at least one binding domain that binds an antigen other than CLEC12a, wherein the at least one binding domain binds CD3, T-cell receptor (TCR)α, TCRβ, CD28, CD16, CD32A, CD64, CD89, NKp46, or NKG2D; and wherein the cancer is selected from chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); acute myeloid leukemia (AML); Hairy cell leukemia; and chronic myeloblastic leukemia.
24. The method of claim 23 , further comprising administering an additional therapy or therapeutic agent.
25. The method of claim 24 , 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.
26. The method of claim 23 , wherein at least one VHH domain that binds CLEC12a comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 32; a CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 34.
27. The method of claim 23 , wherein at least one VHH domain that binds CLEC12a comprises the amino acid sequence of SEQ ID NO: 26 or 94.