PSMA and CD3 bispecific T cell engaging antibody constructs
The present invention provides bispecific antibody constructs of a specific Fc modality characterized by comprising a first domain binding to PSMA, a second domain binding to an extracellular epitope of the human and the Macaca CD3ε chain and a third domain, which is the specific Fc modality. Moreover, the invention provides a polynucleotide, encoding the antibody construct, a vector comprising this polynucleotide, host cells, expressing the construct and a pharmaceutical composition comprising the same.
1. An antibody construct comprising:
a first domain that binds prostate-specific membrane antigen (PSMA) comprising a VH region comprising a CDR-H1 comprising SEQ ID NO: 372, a CDR-H2 comprising SEQ ID NO: 373, and a CDR-H3 comprising SEQ ID NO: 374, and a VL region comprising a CDR-L1 comprising SEQ ID NO: 375, a CDR-L2 comprising SEQ ID NO: 376, and a CDR-L3 comprising SEQ ID NO: 377;
a second domain that binds to an extracellular epitope of the human and Macaca CD3 epsilon (CD3E) chain comprising a VH region comprising a CDR-H1 comprising SEQ ID NO: 544, a CDR-H2 comprising SEQ ID NO: 545, and a CDR-H3 comprising SEQ ID NO: 546, and a VL region comprising a CDR-L1 comprising SEQ ID NO: 514, a CDR-L2 comprising SEQ ID NO: 515, and a CDR-L3 comprising SEQ ID NO: 516; and
a third domain comprising two Fc monomers, each comprising a hinge, a CH2 domain and a CH3 domain, and each Fc monomer comprising the amino acid sequence of SEQ ID NO: 17, wherein the two Fc monomers are fused to each other via a peptide linker.
2. The antibody construct of claim 1 , wherein:
the first domain comprises a VH region comprising at least 90% identity to the amino acid sequence of SEQ ID NO: 378 and a VL region comprising at least 90% identity to the amino acid sequence of SEQ ID NO: 379; and
the second domain comprises a VH region comprising at least 90% identity to the amino acid sequence of SEQ ID NO: 585 and a VL region comprising at least 90% identity to the amino acid sequence of SEQ ID NO: 565.
3. The antibody construct of claim 2 , wherein:
the first domain comprises a VH region comprising the amino acid sequence of SEQ ID NO: 378 and a VL region comprising the amino acid sequence of SEQ ID NO: 379;
the second domain comprises a VH region comprising the amino acid sequence of SEQ ID NO: 585 and a VL region comprising the amino acid sequence of SEQ ID NO: 565; and
the third domain comprises the amino acid sequence of SEQ ID NO: 25.
4. The antibody construct of claim 1 , wherein:
the first domain comprises at least 90% identity to the amino acid sequence of SEQ ID NO: 380; and
the second domain comprises at least 90% identity to the amino acid sequence of SEQ ID NO: 506.
5. The antibody construct of claim 4 , wherein:
the first domain comprises the amino acid sequence of SEQ ID NO: 380;
the second domain comprises the amino acid sequence of SEQ ID NO: 506; and
the third domain comprises the amino acid sequence of SEQ ID NO: 25.
6. The antibody construct of claim 5 , wherein the antibody construct comprises the amino acid sequence of SEQ ID NO: 382.
7. The antibody construct of claim 1 , wherein the antibody construct is a single chain antibody construct.
8. The antibody construct of claim 1 , wherein the antibody construct comprises in an amino to carboxyl order:
the first domain, a first peptide linker, the second domain, a second peptide linker, and the third domain.
9. The antibody construct of claim 1 , wherein the the third domain comprises the amino acid sequence of SEQ ID NO: 25.
10. The antibody construct of claim 1 , wherein the CH2 domain in each Fc monomer comprises an intra domain cysteine disulfide bridge.
11. The antibody construct of claim 1 , wherein the first and second domains are fused to the third domain via a peptide linker.
12. The antibody construct of claim 1 , wherein the antibody construct comprises in an amino to carboxyl order:
(a) the first domain;
(b) a peptide linker having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-3;
(c) the second domain;
(d) a peptide linker having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 9, 10, 11 and 12;
(e) the first Fc monomer of the third domain;
(f) a peptide linker having an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 6, 7 and 8; and
(g) the second Fc monomer of the third domain.
13. The antibody construct of claim 1 , wherein the antibody construct comprises in an amino to carboxyl order:
(a) the first domain comprising the amino acid sequence of SEQ ID NO: 380;
(b) a peptide linker having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-3;
(c) the second domain comprising the amino acid sequence of SEQ ID NO: 506;
(d) a peptide linker having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 9, 10, 11 and 12;
(e) the first Fc monomer of the third domain comprising the amino acid sequence of SEQ ID NO: 17;
(f) a peptide linker having an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 6, 7 and 8; and
(g) the second Fc monomer of the third domain comprising the amino acid sequence of SEQ ID NO: 17.
14. The antibody construct of claim 1 , wherein the first domain and the second domain are each single-chain variable fragments (scFv).
15. The antibody construct of claim 14 , wherein the VH and VL regions in each scFv are arranged in the order VH-VL.
16. A polynucleotide comprising a nucleotide sequence encoding the antibody construct of claim 1 .
17. A vector comprising the polynucleotide of claim 16 .
18. A host cell transformed or transfected with the polynucleotide of claim 16 .
19. A composition comprising the antibody construct of claim 1 and a carrier, stabilizer, excipient, diluent, solubilizer, surfactant, emulsifier, preservative or adjuvant.
20. A method of treating or ameliorating prostate cancer in a subject having prostate cancer expressing PSMA comprising administering to the subject an effective amount of the antibody construct of claim 1 .
21. A method of treating or ameliorating cancer in a subject having a cancer expressing PSMA comprising administering to the subject an effective amount of the antibody construct of claim 1 .
22. A process for producing an antibody construct that binds to PSMA and human and Macaca CD3ε chain, said process comprising culturing a host cell transformed or transfected with the polynucleotide of claim 16 under conditions allowing the expression of the antibody construct, and recovering the antibody construct from the culture.