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US-20260125469-A1 - ANTIBODIES BINDING TO CD3

US20260125469A1US 20260125469 A1US20260125469 A1US 20260125469A1US-20260125469-A1

Abstract

The present invention generally relates to antibodies that bind to CD3, including multispecific antibodies e.g. for activating T cells. In addition, the present invention relates to polynucleotides encoding such antibodies, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the antibodies, and to methods of using them in the treatment of disease.

Inventors

  • Alejandro Carpy Gutierrez Cirlos
  • Anne Freimoser-Grundschober
  • Thomas Hofer
  • Christian Klein
  • Ekkehard Moessner
  • Christiane Neumann
  • Pablo Umaña

Assignees

  • HOFFMANN-LA ROCHE INC.

Dates

Publication Date
20260507
Application Date
20250807
Priority Date
20200619

Claims (20)

  1. 1 . An antibody that binds to CD3, wherein the antibody comprises a first antigen binding domain, comprising (i) a heavy chain variable region (VH) selected from the group consisting of (a) a VH comprising a heavy chain complementary determining region (HCDR) 1 of SEQ ID NO: 2, a HCDR 2 of SEQ ID NO: 4, and a HCDR 3 of SEQ ID NO: 10, (b) a VH comprising a HCDR 1 of SEQ ID NO: 2, a HCDR 2 of SEQ ID NO: 4, and a HCDR 3 of SEQ ID NO: 12, (c) a VH comprising a HCDR 1 of SEQ ID NO: 2, a HCDR 2 of SEQ ID NO: 5, and a HCDR 3 of SEQ ID NO: 9, (d) a VH comprising a HCDR 1 of SEQ ID NO: 3, a HCDR 2 of SEQ ID NO: 6, and a HCDR 3 of SEQ ID NO: 11, or (e) a VH comprising a HCDR 1 of SEQ ID NO: 3, a HCDR 2 of SEQ ID NO: 7, and a HCDR 3 of SEQ ID NO: 13, and (ii) a light chain variable region (VL) comprising a light chain complementarity determining region (LCDR) 1 of SEQ ID NO: 20, a LCDR 2 of SEQ ID NO: 21 and a LCDR 3 of SEQ ID NO: 22.
  2. 2 . The antibody of claim 1 , wherein the VH comprises an amino acid sequence that is at least about 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 15, SEQ ID NO: 17 and SEQ ID NO: 19, and/or the VL comprises an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 23.
  3. 3 . (canceled)
  4. 4 . The antibody of claim 1 , wherein; (a) the first antigen binding domain is a Fab molecule; (b) the antibody comprises an Fc domain composed of a first and a second subunit; and/or (c) the antibody comprises a second antigen binding domain that binds to a second antigen.
  5. 5 . (canceled)
  6. 6 . The antibody of claim 4 , wherein: (a) the antibody comprises a second antigen binding domain that binds to a second antigen, and wherein: (i) the antibody comprises a third antigen binding domain that binds to the second antigen; and/or (ii) the second antigen binding domain is a Fab molecule; and/or (b) the first antigen binding domain is a Fab molecule, and wherein the variable domains VL and VH or the constant domains CL and CH1 are replaced by each other.
  7. 7 . The antibody of claim 6 , wherein: (a) the third antigen binding domain is a Fab molecule; and/or (b) the second antigen binding domain is a conventional Fab molecule.
  8. 8 . (canceled)
  9. 9 . The antibody of claim 7 , wherein the third antigen binding domain is a conventional Fab molecule.
  10. 10 . The antibody of claim 6 , wherein: (a) the second is a Fab molecule wherein in the constant domain CL the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat) and the amino acid at position 123 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the constant domain CH1 the amino acid at position 147 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index) and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index); (b) the first and the second antigen binding domain are fused to each other; (c) the first and the second antigen binding domain are each a Fab molecule and either (i) the second antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding domain, or (ii) the first antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second antigen binding domain; (d) the first and the second antigen binding domain are each a Fab molecule and the antibody comprises an Fc domain composed of a first and a second subunit; and wherein either (i) the second antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding domain and the first antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain, or (ii) the first antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second antigen binding domain and the second antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain; and/or (e) wherein the second antigen is a target cell antigen.
  11. 11 . The antibody of claim 10 , wherein the first and the second antigen binding domain are fused to each other via a peptide linker.
  12. 12 . (canceled)
  13. 13 . The antibody of claim 6 , wherein the antibody comprises a third antigen binding domain that binds to the second antigen, and wherein: (a) the first, the second, and the third antigen binding domain are each a Fab molecule and the antibody comprises an Fc domain composed of a first and a second subunit; and wherein either (i) the second antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding domain and the first antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain, or (ii) the first antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second antigen binding domain and the second antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain; and the third antigen binding domain is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain; (b) the second and the third antigen binding domain is a Fab molecule wherein in the constant domain CL the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat) and the amino acid at position 123 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the constant domain CH1 the amino acid at position 147 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index) and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index); and/or (c) wherein the second antigen is a target cell antigen.
  14. 14 . The antibody of claim 4 , wherein the antibody comprises an Fc domain composed of a first and a second subunit, and wherein the Fc domain: (a) is an IgG Fc domain; (b) is a human Fc domain; (c) comprises a modification promoting the association of the first and the second subunit of the Fc domain; and/or (d) comprises one or more amino acid substitution that reduces binding to an Fc receptor and/or effector function.
  15. 15 - 17 . (canceled)
  16. 18 . The antibody of claim 10 , wherein the second antigen is a tumor cell antigen.
  17. 19 - 30 . (canceled)
  18. 31 . An isolated polynucleotide encoding the antibody of claim 1 .
  19. 32 . A host cell comprising the isolated polynucleotide of claim 31 .
  20. 33 . A method of producing an antibody that binds to CD3, comprising the steps of (a) culturing the host cell of claim 32 under conditions suitable for the expression of the antibody and (b) recovering the antibody.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application is a divisional of U.S. patent application Ser. No. 18/948,718, filed Nov. 15, 2024, which is a divisional of U.S. patent application Ser. No. 18/639,140, filed Apr. 18, 2024, which is a divisional of U.S. patent application Ser. No. 18/480,324, filed Oct. 3, 2023, which is a divisional of U.S. patent application Ser. No. 17/350,457, filed Jun. 17, 2021, which claims priority to European Application No. 20180968.8, filed Jun. 19, 2020, which is incorporated herein by reference in its entirety. SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on Jul. 31, 2025, is named 51177-051005_Sequence_Listing_7_31_25.xml and is 120,166 bytes in size. FIELD OF THE INVENTION The present invention generally relates to antibodies that bind to CD3, including multispecific antibodies e.g. for activating T cells. In addition, the present invention relates to polynucleotides encoding such antibodies, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the antibodies, and to methods of using them in the treatment of disease. BACKGROUND CD3 (cluster of differentiation 3) is a protein complex composed of four subunits, the CD3γ chain, the CD3δ chain, and two CD3ε chains. CD3 associates with the T-cell receptor and the ζ chain to generate an activation signal in T lymphocytes. CD3 has been extensively explored as drug target. Monoclonal antibodies targeting CD3 have been used as immunosuppressant therapies in autoimmune diseases such as type I diabetes, or in the treatment of transplant rejection. The CD3 antibody muromonab-CD3 (OKT3) was the first monoclonal antibody ever approved for clinical use in humans, in 1985. A more recent application of CD3 antibodies is in the form of bispecific antibodies, binding CD3 on the one hand and a tumor cell antigen on the other hand. The simultaneous binding of such an antibody to both of its targets will force a temporary interaction between target cell and T cell, causing activation of any cytotoxic T cell and subsequent lysis of the target cell. For therapeutic purposes, an important requirement that antibodies have to fulfill is sufficient stability both in vitro (for storage of the drug) an in vivo (after administration to the patient). Modifications like asparagine deamidation are typical degradations for recombinant antibodies and can affect both in vitro stability and in vivo biological functions. Given the tremendous therapeutic potential of antibodies, particularly bispecific antibodies for the activation of T cells, there is a need for CD3 antibodies with optimized properties. SUMMARY OF THE INVENTION The present invention provides antibodies, including multispecific (e.g. bispecific) antibodies, that bind to CD3 with good affinity and are resistant to degradation by e.g. asparagine deamidation and thus particularly stable as required for therapeutic purposes. The (multispecific) antibodies provided combine good efficacy (e.g. target cell killing) and producibility with low toxicity (e.g. no T cell activation in the absence of target cells) and favorable pharmacokinetic properties. As is shown herein, the antibodies, including multispecific antibodies, that bind to CD3, provided by the present invention, retain more than about 95% binding activity to CD3 after 2 weeks at pH 7.4, 37° C., relative to the binding activity after 2 weeks at pH 6, −80° C., as determined by surface plasmon resonance (SPR). In one aspect, the invention provides an antibody that binds to CD3, wherein the antibody comprises a first antigen binding domain, comprising (i) a heavy chain variable region (VH) selected from the group consisting of(a) a VH comprising a heavy chain complementary determining region (HCDR) 1 of SEQ ID NO: 2, a HCDR 2 of SEQ ID NO: 4, and a HCDR 3 of SEQ ID NO: 10,(b) a VH comprising a HCDR 1 of SEQ ID NO: 2, a HCDR 2 of SEQ ID NO: 4, and a HCDR 3 of SEQ ID NO: 12,(c) a VH comprising a HCDR 1 of SEQ ID NO: 2, a HCDR 2 of SEQ ID NO: 5, and a HCDR 3 of SEQ ID NO: 9,(d) a VH comprising a HCDR 1 of SEQ ID NO: 3, a HCDR 2 of SEQ ID NO: 6, and a HCDR 3 of SEQ ID NO: 11, or(e) a VH comprising a HCDR 1 of SEQ ID NO: 3, a HCDR 2 of SEQ ID NO: 7, and a HCDR 3 of SEQ ID NO: 13, and(ii) a light chain variable region (VL) comprising a light chain complementarity determining region (LCDR) 1 of SEQ ID NO: 20, a LCDR 2 of SEQ ID NO: 21 and a LCDR 3 of SEQ ID NO: 22. In one aspect, the VH comprises an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 15, SEQ ID NO: 17 and SEQ ID NO: 19, and/or the VL comprises an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical