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EP-4735480-A1 - MULTI-SPECIFIC ANTIBODIES COMPRISING ANTIGEN-BINDING DOMAINS SPECIFICALLY RECOGNIZING C5AR1 AND GM-CSFR ALPHA

EP4735480A1EP 4735480 A1EP4735480 A1EP 4735480A1EP-4735480-A1

Abstract

Provides multi-specific (e.g., bispecific) antibodies that specifically recognize C5aR1 and GM-CSFRα, and pharmaceutical compositions comprising multi-specific (e.g., bispecific) antibodies that specifically recognize C5aR1 and GM-CSFRα. Also provided are methods of making and using these antibodies.

Inventors

  • HE, Chong
  • ZHAI, WENWU
  • ZHOU, ZHIWEN
  • STRAINIC, MICHAEL GEORGE
  • YUAN, PING
  • YI, Zuoan

Assignees

  • Staidson (Beijing) Biopharmaceuticals Co., Ltd.

Dates

Publication Date
20260506
Application Date
20240627

Claims (20)

  1. A multi-specific antibody, which comprises a first antigen-binding domain specifically recognizing C5aR1, and a second antigen-binding domain specifically recognizing GM-CSFRα.
  2. The multi-specific antibody of claim 1, wherein the first antigen-binding domain comprises: (i) a V H comprising an HC-CDR1 comprising SGYSWH (SEQ ID NO: 1) , an HC-CDR2 comprising YIHSSGDTDYNPSLKR (SEQ ID NO: 2) , and an HC-CDR3 comprising SIGDY (SEQ ID NO: 3) ; and a V L comprising an LC-CDR1 comprising RSSQSLVHSNGNTYLH (SEQ ID NO: 11) ; an LC-CDR2 comprising KVSNRFS (SEQ ID NO: 12) , and an LC-CDR3 comprising SQSTHVPWT (SEQ ID NO: 13) ; or (ii) a V H comprising an HC-CDR1 comprising SYWMN (SEQ ID NO: 4) ; an HC-CDR2 comprising RIDPX 1 DSX 2 TRYNQKFED (SEQ ID NO: 49) , wherein X 1 is Y, G or R, and X 2 is E or G; and an HC-CDR3 comprising FVX 1 PX 2 GX 3 FAY (SEQ ID NO: 50) , wherein X 1 is I or W, X 2 is S or V, and X 3 is G or A; and a V L comprising an LC-CDR1 comprising RSSX 1 SPX 2 HSNGNTYLH (SEQ ID NO: 51) , wherein X 1 is Q or R, and X 2 is V or L; an LC-CDR2 comprising KVSNRFS (SEQ ID NO: 17) , and an LC-CDR3 comprising SQSTX 1 VPPT (SEQ ID NO: 52) , wherein X 1 is H or L.
  3. The multi-specific antibody of claim 1 or 2, wherein the first antigen-binding domain comprises: (i) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 9; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 17, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 18; or (ii) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 10; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 16, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 17, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19.
  4. The multi-specific antibody of any one of claims 1-3, wherein the second antigen-binding domain comprises: (i) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42; (ii) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42; or (iii) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43.
  5. The multi-specific antibody of any one of claims 1-4, wherein: (i) the first antigen-binding domain comprises: a V H comprising an HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13, and wherein the second antigen-binding domain comprises: a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42; (ii) the first antigen-binding domain comprises: a V H comprising an HC-CDR1 comprising the amino acid sequence SEQ ID NO: 4, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 9; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 17, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 18, and wherein the second antigen-binding domain comprises: a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42; or (iii) the first antigen-binding domain comprises: a V H comprising an HC-CDR1 comprising the amino acid sequence SEQ ID NO: 4, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 10; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 16, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 17, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and wherein the second antigen-binding domain comprises: a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42.
  6. The multi-specific antibody of any one of claims 1-5, wherein the first antigen-binding domain comprises: (i) a V H comprising the amino acid sequence of SEQ ID NO: 21 or 57, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 21 or 57; and a V L comprising the amino acid sequence of SEQ ID NO: 24 or 60, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 24 or 60; (ii) a V H comprising the amino acid sequence of SEQ ID NO: 22 or 58, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 22 or 58; and a V L comprising the amino acid sequence of SEQ ID NO: 25 or 61, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 25 or 61; (iii) a V H comprising the amino acid sequence of SEQ ID NO: 28 or 64, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 28 or 64; and a V L comprising the amino acid sequence of SEQ ID NO: 32 or 68, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 32 or 68; or (iv) a V H comprising the amino acid sequence of SEQ ID NO: 29 or 65, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 29 or 65; and a V L comprising the amino acid sequence of SEQ ID NO: 33 or 69, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 33 or 69.
  7. The multi-specific antibody of any one of claims 1-6, wherein the second antigen-binding domain comprises: (i) a V H comprising the amino acid sequence of SEQ ID NO: 44 or 70, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 44 or 70; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73; (ii) a V H comprising the amino acid sequence of SEQ ID NO: 45 or 71, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 45 or 71; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73; or (iii) a V H comprising the amino acid sequence of SEQ ID NO: 46 or 72, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 46 or 72; and a V L comprising the amino acid sequence of SEQ ID NO: 48 or 74, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 48 or 74.
  8. The multi-specific antibody of any one of claims 1-7, wherein: (i) the first antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 21 or 57, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 21 or 57; and a V L comprising the amino acid sequence of SEQ ID NO: 24 or 60, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 24 or 60; and wherein the second antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 44 or 70, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 44 or 70; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73; (ii) the first antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 22 or 58, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 22 or 58; and a V L comprising the amino acid sequence of SEQ ID NO: 25 or 61, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 25 or 61; and wherein the second antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 44 or 70, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 44 or 70; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73; (iii) the first antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 28 or 64, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 28 or 64; and a V L comprising the amino acid sequence of SEQ ID NO: 32 or 68, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 32 or 68; and wherein the second antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 44 or 70, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 44 or 70; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73; or (iv) the first antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 29 or 65, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 29 or 65; and a V L comprising the amino acid sequence of SEQ ID NO: 33 or 69, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 33 or 69; and wherein the second antigen-binding domain comprises: a V H comprising the amino acid sequence of SEQ ID NO: 44 or 70, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 44 or 70; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73.
  9. The multi-specific antibody of claim 1, wherein the second antigen-binding domain comprises: (i) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42; (ii) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42; or (iii) a V H comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39; and a V L comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 40, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 41, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43.
  10. The multi-specific antibody of claim 9, wherein the second antigen-binding domain comprises: (i) a V H comprising the amino acid sequence of SEQ ID NO: 44 or 70, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 44 or 70; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73; (ii) a V H comprising the amino acid sequence of SEQ ID NO: 45 or 71, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 45 or 71; and a V L comprising the amino acid sequence of SEQ ID NO: 47 or 73, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 47 or 73; or (iii) a V H comprising the amino acid sequence of SEQ ID NO: 46 or 72, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 46 or 72; and a V L comprising the amino acid sequence of SEQ ID NO: 48 or 74, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 48 or 74.
  11. The multi-specific antibody of any one of claims 1-10, wherein the format of which is selected form the group consisting of DVD-Ig, IgG- (scFv) 2 and Hetero H, CrossMab.
  12. The multi-specific antibody of any one of claims 1-11, comprising four polypeptide chains: wherein: the two polypeptide chains comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V H 1-L-V H 2-C H 1, wherein V H 1 is a heavy-chain variable region that specifically binds to the C5aR1; V H 2 is a heavy chain variable region that specifically binds to GM-CSFRα; L is a linker; C H 1 is a heavy chain constant region C H 1 domain; optionally, wherein the polypeptide chain may further comprise Fc region comprising C H 2 and C H 3 domains; and the other two polypeptide chains comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V L 1-L-V L 2-C L , wherein V L 1 is a light chain variable region that specifically binds to C5aR1; V L 2 is a light chain variable region that specifically binds to GM-CSFRα; L is a linker; and C L is a light chain constant region; wherein V H 1 and V L 1 constitute the first antigen-binding domain specifically recognizing C5aR1 and V H 2 and V L 2 constitute the second antigen-binding domain specifically recognizing GM-CSFRα.
  13. The multi-specific antibody of any one of claims 1-11, comprising four polypeptide chains: wherein: the two polypeptide chains each comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V H 1-L-V H 2-C H 1, wherein V H 1 is a heavy-chain variable region that specifically binds to the GM-CSFRα; V H 2 is a heavy chain variable region that specifically binds to C5aR1; L is a linker; C H 1 is a heavy chain constant region C H 1 domain; optionally, wherein the polypeptide chain may further comprise Fc region comprising C H 2 and C H 3 domains; and the other two polypeptide chains each comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V L 1-L-V L 2-C L , wherein V L 1 is a light chain variable region that specifically binds to GM-CSFRα; V L 2 is a light chain variable region that specifically binds to C5aR1; L is a linker; and C L is a light chain constant region; wherein V H 1 and V L 1 constitute the first antigen-binding domain specifically recognizing GM-CSFRα and V H 2 and V L 2 constitute the second antigen-binding domain specifically recognizing C5aR1.
  14. The multi-specific antibody of claim 12 or 13, which comprises: the amino acid sequence of any one of SEQ ID NOs: 101-102, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of any one of SEQ ID NOs: 101-102; and/or the amino acid sequence of any one of SEQ ID NOs: 79-80, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of any one of SEQ ID NOs: 79-80.
  15. The multi-specific antibody of claim 12 or 13, which comprises: the amino acid sequence of any one of SEQ ID NOs: 103-104, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of any one of SEQ ID NOs: 103-104; and/or the amino acid sequence of any one of SEQ ID NOs: 81-82, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of any one of SEQ ID NOs: 81-82.
  16. The multi-specific antibody of claim 12 or 13, which comprises: (i) the amino acid sequence of SEQ ID NO: 75, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 75; and the amino acid sequence of SEQ ID NO: 79, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 79; (ii) the amino acid sequence of SEQ ID NO: 75, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 75; and the amino acid sequence of SEQ ID NO: 80, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 80; (iii) the amino acid sequence of SEQ ID NOs: 76, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 76; and the amino acid sequence of SEQ ID NO: 79, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 79; (iv) the amino acid sequence of SEQ ID NO: 76, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 76; and the amino acid sequence of SEQ ID NOs: 80, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 80; (v) the amino acid sequence of SEQ ID NO: 77, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 77; and the amino acid sequence of SEQ ID NO: 81, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 81; (vi) the amino acid sequence of SEQ ID NO: 77, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 77; and the amino acid sequence of SEQ ID NO: 82, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 82; (vii) the amino acid sequence of SEQ ID NO: 78, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 78; and the amino acid sequence of SEQ ID NO: 81, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 81; or (viii) the amino acid sequence of SEQ ID NO: 78, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 78; and the amino acid sequence of SEQ ID NO: 82, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 82.
  17. The multi-specific antibody of any one of claims 1-11, comprising four polypeptide chains, wherein: two polypeptide chains each comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V H 1-C H 1-Fc-L1-V H 2-L2-V L 2, or V H 1-C H 1-Fc-L1-V L 2-L2-V H 2, wherein V H 1 is a heavy chain variable region that specifically binds to C5aR1; V H 2 is a heavy chain variable region that specifically binds to GM-CSFRα; V L 2 is a light chain variable region that specifically binds to GM-CSFRα; L1 and L2 are linkers; C H 1 is a heavy chain constant region C H 1 domain; Fc region comprises C H 2 and C H 3 domains; and the other two polypeptide chains each comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V L 1-C L , wherein V L 1 is a light chain variable region that specifically binds to C5aR1, and C L is a light chain constant region; wherein V H 1 and V L 1 constitute the first antigen-binding domain specifically recognizing C5aR1 and V H 2 and V L 2 constitute the second antigen-binding domain specifically recognizing GM-CSFRα.
  18. The multi-specific antibody of any one of claims 1-11, comprising four polypeptide chains, wherein: two polypeptide chains comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V H 1-C H 1-Fc-L1-V H 2-L2-V L 2, or V H 1-C H 1-Fc-L1-V L 2-L2-V H 2, wherein V H 1 is a heavy chain variable region that specifically binds to GM-CSFRα; V H 2 is a heavy chain variable region that specifically binds to C5aR1; V L 2 is a light chain variable region that specifically binds to C5aR1; L1 and L2 are linkers; C H 1 is a heavy chain constant region C H 1 domain; Fc region comprising C H 2 and C H 3 domains; and the other two polypeptide chains comprise polypeptide sequences in the orientation from N-terminus to the C-terminus: V L 1-C L , wherein V L 1 is a light chain variable region that specifically binds to GM-CSFRα, and C L is a light chain constant region; wherein V H 1 and V L 1 constitute the first antigen-binding domain specifically recognizing GM-CSFRα and V H 2 and V L 2 constitute the second antigen-binding domain specifically recognizing C5aR1.
  19. The multi-specific antibody of claim 17 or 18, wherein the V H 2 and V L 2 further comprise cysteines substitution.
  20. The multi-specific antibody of any one of claims 17-19, which comprises: (i) the amino acid sequence of SEQ ID NO: 83, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 83; and/or the amino acid sequence of SEQ ID NO: 87, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 87; (ii) the amino acid sequence of SEQ ID NO: 84, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 84; and/or the amino acid sequence of SEQ ID NO: 88, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 88; (iii) the amino acid sequence of SEQ ID NO: 85, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 85; and/or the amino acid sequence of SEQ ID NO: 87, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 87; or (iv) the amino acid sequence of SEQ ID NO: 86, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 86; and/or the amino acid sequence of SEQ ID NO: 88, or a variant thereof having at least about 90%sequence identity to the amino acid sequence of SEQ ID NO: 88.

Description

MULTI-SPECIFIC ANTIBODIES COMPRISING ANTIGEN-BINDING DOMAINS SPECIFICALLY RECOGNIZING C5aR1 AND GM-CSFR alpha SUBMISSION OF SEQUENCE LISTING ON ASCII TEXT FILE The content of the following submission of sequence list is incorporated herein by reference in its entirety: C5aR1 and GM-CSFRα bispecific antibody seqlist 20230627. xml, date recorded: June 27, 2023, size: 127KB) . FIELD This application relates to multi-specific (e.g., bispecific) antibodies comprising a first antigen-binding domain that specifically recognizing C5aR1 and a second antigen-binding domain that specifically recognizing GM-CSFRα. The application further relates to pharmaceutical compositions comprising antibodies specifically recognizing C5aR1 and antibodies specifically recognizing GM-CSFRα; pharmaceutical compositions comprising the multi-specific (e.g., bispecific) antibody recognizing C5aR1 and GM-CSFRα; as well as methods of manufacture and uses thereof, including methods of treating and preventing inflammatory disease or condition. BACKGROUND Granulocyte-macrophage colony stimulating factor (GM-CSF) is also known as colony-stimulating factor 2 (CSF2) . GM-CSF is a type I proinflammatory cytokine which plays a role in exacerbating inflammatory, respiratory and autoimmune diseases. The GM-CSF receptor is a member of the hematopoietic receptor superfamily. It is heterodimeric, consisting of an alpha and a beta subunit. GM-CSF is able to bind with relatively low affinity to the αsubunit alone (Kd 1-5nM) but not at all to the β subunit alone. However, the presence of both α and β subunits results in a high affinity ligand-receptor complex (Kd≈100pM) . Neutralization of GM-CSF binding to GM-CSFRα is therefore a therapeutic approach to treating diseases and conditions mediated through GM-CSFR. C5a anaphylatoxin chemotactic receptor 1 (C5aR1, also known as CD88) is a member of the rhodopsin superfamily of G-protein-coupled receptor (GPCR) (Gerard NP et al., J Biol Chem 264: 1760-1766, 1989) . C5aR1, with its high affinity ligand C5a, is an essential part of the complement system, playing important role in eliciting the broad immune responses. The structure of C5aR1 conforms to the seven transmembrane receptor family and contains 4 extracellular domains, seven membrane-spanning helices, and 4 intracellular C domains (Dixon et al. Nature 326, 73-77, 1987; Findlay et al. Biochem. J. 238, 625-642, 1986) . C5aR1 is a classical G protein-coupled receptor that signals through Gαi and Gα16 which both are G- proteins. Activated C5aR1 has been shown to associate with two of the four mammalian β-arrestins (β-arrestin 1, 2) , which have different dependencies on the phosphorylation status of the receptor. Upon agonist biding, C5aR1 undergoes rapid phosphorylation on the six serine residues present in the C-terminal region followed by desensitization and internalization. Serine residues in the C5aR1 C terminus were identified that control the intracellular trafficking of the C5aR1-arrestin complex in response to C5a. (Braun et al. J Biol Chem. 278: 4277-4285, 2003) . C5aR1 has been found to be expressed on various inflammatory cells and smooth muscle cells to promote an inflammatory reaction. It is also present on subsets of T cells and dendritic cells to regulate the adaptive immune response (Kemper et al. Nature Rev. Immunol. 7.9-18, 2007) . C5a-C5aR1 interaction is required for the optimal oxidative burst and phagocytosis, which form the fundamental machinery in innate immunity for bacterial clearance. The binding of C5a ligand and its receptor C5aR1 thereof induces an inflammatory reaction by inducing cytokine and chemokine expression (Monsinjon et al. FASEB J 17: 1003–1014, 2003.; Fukuoka et al. Clin Exp Immunol 131: 248–253, 2003) . C5a-C5aR1 interaction is also an essential step in neutrophil migration by causing adhesion molecule expression and inducing chemokine production (Riedemann et al. Immunity 19: 193–202, 2003. ) . It has been reported that C5aR1 and its ligand C5a play a key role in initiating and maintaining several inflammatory responses by recruiting and activating neutrophils and monocytes in the lungs (Julien Carvelli et al. Nature 588, 146-150, 2020) . In addition, C5a perturbs coagulation and fibrinolysis pathway, resulting in damaging intravascular coagulation (Muhlfelder et al. J Clin Invest 63: 147–150, 1979.; Laudes et al. Am J Pathol 160: 1867–1875, 2002. ) . In addition, it has been reported that activated C5aR1 and C3aR signal through the PI3K/AKT pathway in cancer cells, and silencing the PI3K or AKT gene in cancer cells eliminates the progrowth effects of C5aR1 and C3aR stimulation. In patients with ovarian or lung cancer, higher tumoral C3 or C5aR1 mRNA levels were associated with decreased overall survival (Cho Ms et al. Cell Rep. 2014) . Inhibition of C5a-C5aR1 interaction should reduce the acute inflammatory response mediated via C5a. The blockade of C5aR1 signal, thereby reducing of the inflammatory rea