Bispecific T cell activating antigen binding molecules

Patent metadata

Field	Value
Publication number	US-11117965-B2
Application number	US-202016799662-A
Country	US
Kind code	B2
Filing date	Feb 24, 2020
Priority date	Aug 4, 2014
Publication date	Sep 14, 2021
Grant date	Sep 14, 2021

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 first polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 18, a second polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 19, a third polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 20, and a fourth polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 21, wherein the T cell activating bispecific antigen-binding molecule comprises:
(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 123 and 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. A T cell activating bispecific antigen-binding molecule comprising a first polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 18, a second polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 19, a third polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 20, and a fourth polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 21, wherein the T cell activating bispecific antigen-binding molecule comprises:
(a) a first Fab molecule which specifically binds to CD20, wherein the first Fab molecule comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 30 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 31;
(b) a second Fab molecule which specifically binds to CD3, wherein the second Fab molecule comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, 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 a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 30 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 31, 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).

3. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein the first polypeptide sequence is at least 96% identical to the amino acid sequence of SEQ ID NO: 18, the second polypeptide sequence is at least 96% identical to the amino acid sequence of SEQ ID NO: 19, the third polypeptide sequence is at least 96% identical to the amino acid sequence of SEQ ID NO: 20, and/or the fourth polypeptide sequence is at least 96% identical to the amino acid sequence of SEQ ID NO: 21.

4. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein the first polypeptide sequence is at least 97% identical to the amino acid sequence of SEQ ID NO: 18, the second polypeptide sequence is at least 97% identical to the amino acid sequence of SEQ ID NO: 19, the third polypeptide sequence is at least 97% identical to the amino acid sequence of SEQ ID NO: 20, and/or the fourth polypeptide sequence is at least 97% identical to the amino acid sequence of SEQ ID NO: 21.

5. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein the first polypeptide sequence is at least 98% identical to the amino acid sequence of SEQ ID NO: 18, the second polypeptide sequence is at least 98% identical to the amino acid sequence of SEQ ID NO: 19, the third polypeptide sequence is at least 98% identical to the amino acid sequence of SEQ ID NO: 20, and/or the fourth polypeptide sequence is at least 98% identical to the amino acid sequence of SEQ ID NO: 21.

6. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein the first polypeptide sequence is at least 99% identical to the amino acid sequence of SEQ ID NO: 18, the second polypeptide sequence is at least 99% identical to the amino acid sequence of SEQ ID NO: 19, the third polypeptide sequence is at least 99% identical to the amino acid sequence of SEQ ID NO: 20, and/or the fourth polypeptide sequence is at least 99% identical to the amino acid sequence of SEQ ID NO: 21.

7. The T cell activating bispecific antigen-binding molecule of claim 1 , wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 18, the second polypeptide comprises the amino acid sequence of SEQ ID NO: 19, the third

Assignees

• Hoffmann La Roche

Inventors

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

Classifications

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/732

  Antibody-dependent cellular cytotoxicity [ADCC] · CPC title

• C07K2317/71

  Decreased effector function due to an Fc-modification · CPC title

• C07K2317/66

  comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1 · CPC title

• C07K2317/55

  Fab or Fab' · CPC title