Optimized cross-species specific bispecific single chain antibody contructs

Patent metadata

Field	Value
Publication number	US-11661462-B2
Application number	US-201515329672-A
Country	US
Kind code	B2
Filing date	Jul 31, 2015
Priority date	Jul 31, 2014
Publication date	May 30, 2023
Grant date	May 30, 2023

Abstract

The present invention provides to a bispecific single chain antibody construct binding to a target cell surface antigen via a first binding domain and to the T cell surface antigen CD3 via a second binding domain, wherein serum albumin is fused to the C-terminus of the antibody construct. Moreover, the invention provides a polynucleotide encoding the antibody construct, a vector comprising said polynucleotide and a host cell transformed or transfected with said vector. Furthermore, the invention provides a process for the production of the antibody construct of the invention, a medical use of said antibody construct and a kit comprising said antibody construct.

First claim

The invention claimed is:

1. A bispecific single chain antibody construct comprising a first binding domain which binds a viral antigen on the surface of an infected host cell and a second binding domain which binds to a T cell surface antigen CD3, wherein the second binding domain comprises a VL region having CDR-L1-L3 and a VH region having CDR-H1-H3 selected from the group consisting of:
(a) CDR-L1-L3 as depicted in SEQ ID NOs: 14-16 and CDR-H1-H3 as depicted in SEQ ID NOs: 17-19;
(b) CDR-L1-L3 as depicted in SEQ ID NOs: 26-28 and CDR-H1-H3 as depicted in SEQ ID NOs: 29-31;
(c) CDR-L1-L3 as depicted in SEQ ID NOs: 38-40 and CDR-H1-H3 as depicted in SEQ ID NOs: 41-43;
(d) CDR-L1-L3 as depicted in SEQ ID NOs: 50-52 and CDR-H1-H3 as depicted in SEQ ID NOs: 53-55;
(e) CDR-L1-L3 as depicted in SEQ ID NOs: 62-64 and CDR-H1-H3 as depicted in SEQ ID NOs: 65-67;
(f) CDR-L1-L3 as depicted in SEQ ID NOs: 74-76 and CDR-H1-H3 as depicted in SEQ ID NOs: 77-79;
(g) CDR-L1-L3 as depicted in SEQ ID NOs: 86-88 and CDR-H1-H3 as depicted in SEQ ID NOs: 89-91;
(h) CDR-L1-L3 as depicted in SEQ ID NOs: 98-100 and CDR-H1-H3 as depicted in SEQ ID NOs: 101-103;
(i) CDR-L1-L3 as depicted in SEQ ID NOs: 110-112 and CDR-H1-H3 as depicted in SEQ ID NOs: 113-115; and
(j) CDR-L1-L3 as depicted in SEQ ID NOs: 122-124 and CDR-H1-H3 as depicted in SEQ ID NOs: 125-127;

and
wherein a serum albumin is fused to the C-terminus of the construct to sterically impair dimerization or multimerization of the antibody construct via the second binding domain to prevent activation of T cells in the absence of target cells up to a concentration of 2 mg/ml of the bispecific single chain antibody construct.

2. The antibody construct according to claim 1 , wherein the serum albumin is a human serum albumin or an FcRn binding optimized variant thereof.

3. The antibody construct according to claim 1 , wherein the serum albumin comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 12 and 608 to 628.

4. The antibody construct according to claim 1 , wherein the serum albumin is linked to the antibody construct via a peptide linker.

5. The antibody construct according to claim 4 , wherein the peptide linker has the amino acid sequence (GGGGS) n (SEQ ID NO: 13) n wherein n is an integer in the range of 1 to 5.

6. The antibody construct according to claim 1 , wherein the second binding domain comprises pairs of VH and VL chains selected from the group consisting of:
(a) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 20 and a VL-chain comprising the amino acid sequence set forth in-SEQ ID NO: 22;
(b) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 32 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 34;
(c) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 44 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 46;
(d) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 56 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 58;
(e) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 68 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 70;
(f) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 80 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 82;
(g) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 92 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 94;
(h) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 104 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 106;
(i) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 116 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 118; and
(j) a VH-chain comprising the amino acid sequence set forth in SEQ ID NO: 128 and a VL-chain comprising the amino acid sequence set forth in SEQ ID NO: 130.

7. The antibody construct according to claim 1 , wherein the second binding domain comprises the amino acid sequence set forth in SEQ ID NO: 24, SEQ ID NO: 36, SEQ ID NO: 48, SEQ ID NO: 60, SEQ ID NO: 72, SEQ ID NO: 84, SEQ ID NO: 96, SEQ ID NO: 108, SEQ ID NO: 120 or SEQ ID NO: 132.

8. The antibody construct according to claim 1 , wherein the antibody construct comprises the following elements starting from the N-terminus:
(a) an scFv binding to the viral antigen comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 2654, 2664, 2674, 2684, 2694, and 2704;
(b) a peptide linker comprising the amino acid sequence selected from the group consisting of: SEQ ID NOs: 13 and 2707-2709;
(c) an scFv binding to the T cell surface antigen CD3 comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 36, 48, 60, 72, 84, 96, 108,120,132 and 2706;
(d) a peptide linker comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 13 and 2707-2709;
(e) a serum albumin comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 12 and 608 to 628; and
(f) optionally a His-tag.

9. A pharmaceutical composition comprising the antibody construct according to claim 1 .

10. A kit comprising the antibody construct according to claim 1 and a recipient and, optionally, directions for use.

11. The antibody construct according to claim 1 , wherein the serum albumin is fused to prevent activation of T cells in the absence of target cells at a concentration up to 1 mg/ml of the bispecific single chain antibody construct.

12. The antibody construct according to claim 1 , wherein the serum albumin is fused to prevent activation of T cells in the absence of target cells at a concentration up to 500 ng/ml of the bispecific single chain antibody construct.

Assignees

• Amgen Res Munich Gmbh

Inventors

• Rattel Benno
• Friedrich Matthias
• Kufer Peter
• Hoffmann Patrick
• Raum Tobias
• Münz Markus
• Herrmann Ines
• LUTTERBÜSE Ralf
• Nahrwold Elisabeth

Classifications

• A61K2039/505

  comprising antibodies · CPC title

• C07K16/2809

  against the T-cell receptor (TcR)-CD3 complex · CPC title

• C07K2317/73

  Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• C07K2317/31

  multispecific · CPC title