Tandem diabody for CD16A-directed NK-cell engagement
The invention relates to a multispecific antigen-binding molecule specifically binding to CD16A and consisting of two polypeptide chains, wherein each polypeptide chain comprises at least four variable domains from the N-terminus to the C-terminus in the order: VH_BCMA-VL_CD16A-VH_CD16A-VL_BCM.
1. A dimeric multispecific antigen-binding molecule specifically binding to CD16A and a target cell antigen different from CD16A consisting of two polypeptide chains, wherein each polypeptide chain comprises at least four variable domains selected from the group consisting of:
(i) a heavy chain variable domain specific for CD16A (VH_CD16A) comprising a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1; a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2; a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3,
(ii) a light chain variable domain specific for CD16A (VL_CD16A) comprising a light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 4; a light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 5; and a light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 6,
(iii) a heavy chain variable domain specific for the target cell antigen (VH_TA), and
(iv) a light chain variable domain specific for the target cell antigen (VL_TA),
wherein
these variable domains are linked one after another by peptide linkers L1, L2 and L3 consisting of 12 or less amino acid residues and positioned within each of the two polypeptide chains from the N-terminus to the C-terminus in the order:
VH_TA-L1-VL_CD16A-L2-VH_CD16A-L3-VL_TA.
2. The multispecific antigen-binding molecule of claim 1 , wherein linker L2 consists of less amino acid residues than each of linkers L1 and L3.
3. The multispecific antigen-binding molecule of claim 1 , wherein linker L2 consists of 3 to 9 amino acid residues.
4. The antigen-binding molecule of claim 3 , wherein (i) linker L2 consists of 3 amino acid residues and each of linkers L1 and L3 consists of 6 to 12 amino acid residues or (ii) linker L2 consists of 6 amino acid residues and each of linkers L1 and L3 consists of 9 to 12 amino acid residues.
5. The multispecific antigen-binding molecule of claim 4 , wherein (i) linker L1 consists of 12 amino acid residues, linker L2 consists of 3 amino acid residues and linker L3 consists of 12 amino acid residues or (ii) linker L1 consists of 9 amino acid residues, linker L2 consists of 6 amino acid residues and linker L3 consists of 9 amino acid residues.
6. The multispecific antigen-binding molecule of claim 1 , wherein the target cell antigen is selected from the group consisting of BCMA, CS-1, CD19, CD20, CD38, and CD138.
7. The multispecific antigen-binding molecule of claim 1 , wherein
(i) the heavy chain variable domain specific for CD16A comprises the amino acid sequence set forth in SEQ ID NO:8 (VH_CD16A), and
(ii) the light chain variable domain specific for CD16A comprises the amino acid sequence set forth in SEQ ID NO:9 (VL_CD16A).
8. A polynucleotide encoding a multispecific antigen-binding molecule of claim 1 .
9. A vector comprising a polynucleotide of claim 8 .
10. A host cell transfected with a vector of claim 9 .
11. A method of treating multiple myeloma, comprising administering to a subject in need thereof the multispecific antigen-binding molecule of claim 6 , wherein the target cell antigen is BCMA.