Anti-taa/anti-cd3 multispecific antibody

Patent metadata

Field	Value
Publication number	US-2025304706-A1
Application number	US-202318864299-A
Country	US
Kind code	A1
Filing date	May 11, 2023
Priority date	May 12, 2022
Publication date	Oct 2, 2025
Grant date	—

Abstract

An object of the present invention is to provide an anti-TAA/anti-CD3 multispecific antibody that can be used in treatment of humans. In order to produce an anti-TAA/anti-CD3 multispecific antibody targeting various tumor associated antigens (TAAs), the present inventors utilized known information on anti-CD3 scFv and their own techniques for analyzing protein structure information to obtain 16 types of anti-CD3 scFv having affinity to various CD3 and improved stability. Furthermore, a multispecific antibody combining a plurality of anti-TAA antibodies such as TSPAN8 and CD37 was produced to confirm that the obtained multispecific antibody maintained various binding activities to CD3 and stability. Thus, the obtained 16 types of anti-CD3 scFv can be used in production of an anti-TAA/anti-CD3 multispecific antibody for an arbitrary TAA.

First claim

1 . A multispecific antibody comprising: a heavy chain variable region and a light chain variable region of an anti-tumor associated antigen (TAA) antibody that binds to TAA; and an anti-CD3 scFv region comprising a heavy chain variable region and a light chain variable region of an anti-CD3 antibody that binds to CD3, wherein the anti-CD3 scFv region is any one of:
(1) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 14; (2) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 16; (3) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 18; (4) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 20; (5) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 22; (6) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 24; (7) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 26; (8) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 28; (9) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 30; (10) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 32; (11) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 34; (12) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 36; (13) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 38; (14) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 40; (15) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 42; and (16) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 44.

2 . The multispecific antibody according to claim 1 , wherein the multispecific antibody is a bispecific antibody comprising:
(a) a Fab region of an anti-TAA antibody consisting of a heavy chain fragment comprising a heavy chain variable region of the anti-TAA antibody and a light chain comprising a light chain variable region of the anti-TAA antibody; (b) an anti-CD3 scFV region comprising a heavy chain variable region and a light chain variable region of an anti-CD3 antibody; and (c) an Fc region consisting of a first Fc polypeptide linked to the heavy chain fragment of the Fab region (a) and a second Fc polypeptide linked to the anti-CD3 scFv region (b).

3 . The multispecific antibody according to claim 2 , wherein the Fc region comprises knobs-into-holes mutation or LALA mutation, L234A and L235A, where positions of the mutation are amino acid positions in human Igγ1 constant region according to EU index.

4 . The multispecific antibody according to claim 2 , wherein the Fc region comprises knobs-into-holes mutation and LALA mutation.

5 . The multispecific antibody according to claim 2 , wherein the Fc region comprises P329A mutation and/or P331G mutation, where a position of the mutation is an amino acid position in human Igγ1 constant region according to EU index.

6 . The multispecific antibody according to claim 2 , wherein the heavy chain fragment of the Fab region of the anti-TAA antibody and the first Fc polypeptide are linked via a hinge region; and the anti-CD3 scFv region and the second Fc polypeptide are linked via a hinge region.

7 . The multispecific antibody according to claim 6 , wherein the hinge region linking the anti-CD3 scFv region to the second Fc polypeptide comprises C220S mutation, where a position of the mutation is an amino acid position in human Igγ1 constant region according to EU index.

8 . The multispecific antibody according to claim 6 , wherein the Fc region comprises knobs-into-holes mutation, LALA mutation, and P329A and/or P331G mutation, wherein the hinge region linking anti-CD3 scFv region to the second Fc polypeptide comprises C220S mutation, where a position of the mutation is an amino acid position in human Igγ1 constant region according to EU index.

9 . A multispecific antibody which is an anti-tumor associated antigen (TAA)/anti-CD3 bispecific antibody, comprising: a heavy chain of an anti-TAA antibody that binds to TAA, in which a first Fc polypeptide is linked to a heavy chain fragment comprising a heavy chain variable region of the anti-TAA antibody; a light chain comprising a light chain variable region of the anti-TAA antibody; and a polypeptide in which an anti-CD3 scFv region comprising a heavy chain variable region and a light chain variable region of an anti-CD3 antibody that binds to CD3 and a second Fc polypeptide are linked, wherein the anti-CD3 scFv region is any one of:
(1) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 14; (2) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 16; (3) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 18; (4) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 20; (5) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 22; (6) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 24; (7) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 26; (8) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 28; (9) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 30; (10) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 32; (11) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 34; (12) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 36; (13) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 38; (14) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 40; (15) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 42; and (16) an anti-CD3 scFv region consisting of an amino acid sequence at amino acid positions 1 to 254 of SEQ ID NO: 44.

10 . A multispecific antibody which is a trispecific antibody obtained by further adding an anti-CD137 scFv region comprising a heavy chain variable region and a light chain variable region of an anti-CD137 antibody that binds to CD137 to the multispecific antibody according to claim 2 , wherein the anti-CD137 scFv region is any one of (a) to (e):
(a) an anti-CD137 scFv region consisting of an amino acid sequence at amino acid positions 1 to 242 of SEQ ID NO: 46; (b) an anti-CD137 scFv region consisting of an amino acid sequence at amino acid positions 1 to 244 of SEQ ID NO: 48; (c) an anti-CD137 scFv region consisting of an amino acid sequence at amino acid positions 1 to 249 of SEQ ID NO: 50; (d) an anti-CD137 scFv regio

Assignees

• Astellas Pharma Inc

Inventors

• Soga Shinji
• Yagi Shigenori
• Satake Yoshiki
• Honda Takashi
• Sato Masahito
• Chaen Takashi

Classifications

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K2317/55

  Fab or Fab' · CPC title

• C07K2317/53

  Hinge · CPC title

• C07K2317/31

  multispecific · CPC title

• C07K16/2809 Primary

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