Bispecific T cell activating antigen binding molecules

Patent metadata

Field	Value
Publication number	US-9914776-B2
Application number	US-201514816252-A
Country	US
Kind code	B2
Filing date	Aug 3, 2015
Priority date	Aug 4, 2014
Publication date	Mar 13, 2018
Grant date	Mar 13, 2018

Abstract

The present invention generally relates to novel bispecific antigen binding molecules for T cell activation and re-direction to specific target cells. In addition, the present invention relates to polynucleotides encoding such bispecific antigen binding molecules, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the bispecific antigen binding molecules of the invention, and to methods of using these bispecific antigen binding molecules in the treatment of disease.

First claim

We claim:

1. A T cell activating bispecific antigen-binding molecule comprising:
(a) a first Fab molecule which specifically binds to CD20, wherein the first Fab molecule comprises a heavy chain complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 46, a heavy chain CDR 2 comprising the amino acid sequence of SEQ ID NO: 47, a heavy chain CDR 3 comprising the amino acid sequence of SEQ ID NO: 48, a light chain CDR 1 comprising the amino acid sequence of SEQ ID NO: 49, a light chain CDR 2 comprising the amino acid sequence of SEQ ID NO: 50, and a light chain CDR 3 comprising the amino acid sequence of SEQ ID NO: 51;
(b) a second Fab molecule which specifically binds to CD3, wherein the second Fab molecule comprises a heavy chain CDR 1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR 2 comprising the amino acid sequence of SEQ ID NO: 5, a heavy chain CDR 3 comprising the amino acid sequence of SEQ ID NO: 6, a light chain CDR 1 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR 2 comprising the amino acid sequence of SEQ ID NO: 9, and a light chain CDR 3 comprising the amino acid sequence of SEQ ID NO: 10, wherein the variable domains VL and VH of the Fab light chain and the Fab heavy chain of the second Fab molecule, respectively, are replaced by each other, and
(c) a third Fab molecule which specifically binds to CD20, wherein the third Fab molecule comprises a heavy chain CDR 1 comprising the amino acid sequence of SEQ ID NO: 46, a heavy chain CDR 2 comprising the amino acid sequence of SEQ ID NO: 47, a heavy chain CDR 3 comprising the amino acid sequence of SEQ ID NO: 48, a light chain CDR 1 comprising the amino acid sequence of SEQ ID NO: 49, a light chain CDR 2 comprising the amino acid sequence of SEQ ID NO: 50, and a light chain CDR 3 comprising the amino acid sequence of SEQ ID NO: 51, and wherein:
(i) in the constant domain CL of the first Fab molecule and the third Fab molecule, the amino acid at position 124, according to Kabat, is substituted by lysine (K), arginine (R), or histidine (H), and, in the constant domain CH1 of the first Fab molecule and the third Fab molecule, one or both of the amino acids at positions 147 and 213, according to the Kabat EU index, are substituted by glutamic acid (E) or aspartic acid (D); or
(ii) in the constant domain CL of the second Fab molecule, the amino acid at position 124, according to Kabat, is substituted by lysine (K), arginine (R), or histidine (H), and, in the constant domain CH1 of the second Fab molecule, one or both of the amino acids at positions 147 and 213, according to the Kabat EU index, are substituted by glutamic acid (E) or aspartic acid (D).

2. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein, in the constant domain CL of the first Fab molecule, the amino acid at position 124, according to Kabat, is substituted by lysine (K), arginine (R), or histidine (H); and, in the constant domain CH1 of the first Fab molecule, one or both of the amino acids at positions 147 and 213, according to the Kabat EU index, are substituted by glutamic acid (E) or aspartic acid (D).

3. The T cell activating bispecific antigen-binding molecule of claim 2 , wherein, in the constant domain CH1 of the first Fab molecule, the amino acid at position 147, according to the Kabat EU index, is substituted by glutamic acid (E) or aspartic acid (D).

4. The T cell activating bispecific antigen-binding molecule of claim 3 , wherein, the amino acid at position 123, according to Kabat, is substituted by lysine (K), arginine (R), or histidine (H) in the constant domain CH1 of the first Fab molecule, and the amino acid at position 213, according to the Kabat EU index, is substituted by glutamic acid (E) or aspartic acid (D).

5. The T cell activating bispecific antigen-binding molecule of claim 4 , wherein, in the constant domain CL of the first Fab molecule, the amino acid at position 124, according to Kabat, is substituted by lysine (K), and the amino acid at position 123, according to Kabat, is substituted by arginine (R); and in the constant domain CH1 of the first Fab molecule, the amino acid at position 147, according to the Kabat EU index, is substituted by glutamic acid (E), and the amino acid at position 213, according to the Kabat EU index, is substituted by glutamic acid (E).

6. The T cell activating bispecific antigen-binding molecule of claim 4 , wherein, in the constant domain CL of the first Fab molecule, the amino acid at position 124, according to Kabat, is substituted by lysine (K), and the amino acid at position 123, according to Kabat, is substituted by lysine (K), and in the constant domain CH1 of the first Fab molecule, the amino acid at position 147, according to the Kabat EU index, is substituted by glutamic acid (E), and the amino acid at position 213, according to the Kabat EU index, is substituted by glutamic acid (E).

7. The T cell activating bispecific antigen-binding molecule of claim 1 , further comprising:
(d) an Fc domain comprising a first and a second subunit capable of stable association.

8. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein the first and the second Fab molecule are fused to each other.

9. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein (i) the C-terminus of the Fab heavy chain of the second Fab molecule is fused to the N-terminus of the Fab heavy chain of the first Fab molecule; or (ii) the C-terminus of the Fab heavy chain of the first Fab molecule is fused to the N-terminus of the Fab heavy chain of the second Fab molecule.

10. The T cell activating bispecific antigen-binding molecule of claim 9 , wherein the Fab light chain of the first Fab molecule and the Fab light chain of the second Fab molecule are fused to each other.

11. The T cell activating bispecific antigen-binding molecule of claim 7 , wherein (i) the C-terminus of the Fab heavy chain of the second Fab molecule is fused to the N-terminus of the first or the second subunit of the Fc domain; (ii) the C-terminus of the Fab heavy chain of the first Fab molecule is fused to the N-terminus of the first or the second subunit of the Fc domain; (iii) the C-terminus of the Fab heavy chain of the first and the second Fab molecule are each fused to the N-terminus of one of the subunits of the Fc domain; and/or (iv) the C-terminus of the Fab heavy chain of the third Fab molecule is fused to the N-terminus of the first or second subunit of the Fc domain.

12. The T cell activating bispecific antigen-binding molecule of claim 7 , wherein the C-terminus of the Fab heavy chains of the second and the third Fab molecules are each fused to the N-terminus of one of the subunits of the Fc domain, and the C-terminus of the Fab heavy chain of the first Fab molecule is fused to the N-terminus of the Fab heavy chain of the second Fab molecule.

13. The T cell activating bispecific antigen-binding molecule of claim 7 , wherein the C-terminus of the Fab heavy chain of the first and the third Fab molecules are each fused to the N-terminus of one of the subunits of the Fc domain, and the C-terminus of the Fab heavy chain of the second Fab molecule is fused to the N-terminus of the Fab heavy chain of the first Fab molecule.

14. The T cell activating bispecific antigen-binding molecule of claim 13 , wherein the first and the third Fab molecules and the Fc domain are part of an immunoglobulin molecule.

15. A T cell activating bispecific antigen-binding molecule comprising:
(a) a first Fab molecule which specifically binds to CD20, wherein the first Fab molecule

Assignees

• Hoffmann La Roche

Inventors

• Ast Oliver
• Bacac Marina
• Imhof-Jung Sabine
• Jaeger Christiane
• Klein Christian
• Klostermann Stefan
• Molhoj Michael
• Regula Joerg Thomas
• Schaefer Wolfgang
• Umana Pablo

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K16/2878

  against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 · CPC title

• C07K2317/732

  Antibody-dependent cellular cytotoxicity [ADCC] · CPC title

• C07K2317/51

  Complete heavy chain or Fd fragment, i.e. VH + CH1 · CPC title

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title