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US-12617854-B2 - Humanized or chimeric CD3 antibodies

US12617854B2US 12617854 B2US12617854 B2US 12617854B2US-12617854-B2

Abstract

The present invention relates to humanized or chimeric antibodies binding CD3. It furthermore relates to bispecific antibodies, compositions, pharmaceutical compositions, use of said antibodies in the treatment of a disease, and method of treatment.

Inventors

  • Rik RADEMAKER
  • Isil Altintas
  • Patrick Engelberts
  • Janine Schuurman
  • Paul Parren

Assignees

  • GENMAB A/S

Dates

Publication Date
20260505
Application Date
20220509
Priority Date
20150715

Claims (20)

  1. 1 . A nucleic acid, or set of nucleic acids, encoding a humanized or chimeric antibody which binds to human CD3, wherein said antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein: a) the VH region comprises the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 1, 2, and 176, respectively, and the VL region comprises the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NO: 6, the sequence GTN, and SEQ ID NO: 7, respectively; b) the VH region comprises the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 1, 2, and 184, respectively, and the VL region comprises the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NO: 6, the sequence GTN, and SEQ ID NO: 7, respectively; c) the VH region comprises the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 1, 2, and 220, respectively, and the VL region comprises the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NO: 6, the sequence GTN, and SEQ ID NO: 7, respectively; d) the VH region comprises the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 1, 2, and 236, respectively, and the VL region comprises the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NO: 6, the sequence GTN, and SEQ ID NO: 7, respectively; or e) the VH region comprises the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 1, 2, and 244, respectively, and the VL region comprises the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NO: 6, the sequence GTN, and SEQ ID NO: 7, respectively.
  2. 2 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody comprises a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 1, 2, and 176, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NO: 6, the sequence GTN, and SEQ ID NO: 7, respectively.
  3. 3 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody is a full-length antibody.
  4. 4 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody is chimeric or humanized.
  5. 5 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody is of an isotype selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.
  6. 6 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody comprises: a) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 177 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 8, b) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 185 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 8, c) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 221 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 8, d) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 237 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 8, or e) a VH region comprising the amino acid sequence set forth in SEQ ID NO: 245 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 8.
  7. 7 . The nucleic acid, or set of nucleic acids, according to claim 6 , wherein the antibody comprises a VH region comprising the amino acid sequence set forth in SEQ ID NO: 177 and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 8.
  8. 8 . An expression vector, or set of expression vectors, comprising the nucleic acid, or set of nucleic acids, according to claim 6 .
  9. 9 . A host cell comprising the expression vector, or set of expression vectors, according to claim 8 .
  10. 10 . A host cell comprising the nucleic acid, or set of nucleic acids, according to claim 6 .
  11. 11 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody is monovalent, bivalent or multivalent.
  12. 12 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody comprises a first heavy chain and a second heavy chain.
  13. 13 . The nucleic acid, or set of nucleic acids, according to claim 1 , wherein the antibody comprises a first heavy chain and a second heavy chain, wherein in at least one of said first and second heavy chains, one or more amino acids in the positions corresponding to positions L234, L235, D265, N297, and P331 in a human IgG1 heavy chain according to EU numbering, are not L, L, D, N, and P, respectively.
  14. 14 . The nucleic acid, or set of nucleic acids, according to claim 12 , wherein in at least one of said first and second heavy chains of the antibody, (a) the amino acid in the position corresponding to position D265 in a human IgG1 heavy chain is not D; (b) the amino acid in the position corresponding to position N297 in a human IgG1 heavy chain is not N; (c) the amino acids in the positions corresponding to positions L234 and L235 in a human IgG1 heavy chain are not L and L, respectively; (d) the amino acids in the positions corresponding to positions L234 and L235 in a human IgG1 heavy chain are F and E; or A and A, respectively; (e) the amino acids in the positions corresponding to positions L234 and L235 in a human IgG1 heavy chain are F and E, respectively; (f) the amino acids in the positions corresponding to positions L234 and L235 in a human IgG1 heavy chain are A and A, respectively; (g) the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain are not L, L, and D, respectively; (h) the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain are F, E, and A; or A, A, and A, respectively; (i) the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain are F, E, and A, respectively; (j) the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain, are A, A, and A, respectively; or (k) the amino acids in the positions corresponding to positions L234, L235, D265, N297, and P331 in a human IgG1 heavy chain are F, E, A, Q, and S, respectively.
  15. 15 . The nucleic acid, or set of nucleic acids, according to claim 12 , wherein each of said first and second heavy chain of the antibody comprises at least a hinge region, a CH2 region, and a CH3 region, wherein in said first heavy chain, at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and in said second heavy chain, at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and wherein said first and said second heavy chains are not substituted in the same positions.
  16. 16 . The nucleic acid, or set of nucleic acids, according to claim 15 , wherein (a) the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said first heavy chain, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said second heavy chain, or (b) the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said second heavy chain, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said first heavy chain.
  17. 17 . An expression vector, or set of expression vectors, comprising the nucleic acid, or set of nucleic acids, according to claim 1 .
  18. 18 . A host cell comprising the expression vector, or set of expression vectors, according to claim 17 .
  19. 19 . A host cell comprising the nucleic acid, or set of nucleic acids, according to claim 1 .
  20. 20 . The host cell according to claim 19 , wherein the host cell is a recombinant eukaryotic, recombinant prokaryotic, or recombinant microbial host cell.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a division of application Ser. No. 15/744,317 filed Jan. 12, 2018, which is a 35 U.S.C. 371 national stage filing of International Application No. PCT/EP2016/066845, filed Jul. 14, 2016, which claims priority to International Application No. PCT/EP2016/050296, filed Jan. 8, 2016, which claims priority to Danish Patent Application Nos. PA 2015 00413 filed Jul. 15, 2015, PA 2015 00414 filed Jul. 15, 2015, and PA 2015 00416 filed Jul. 16, 2015. The entire contents of these applications are incorporated herein by reference in their entirety. SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on May 9, 2022, is named GMI-147USFBDV_SequenceListing_20220509.txt and is 298,446 bytes in size. FIELD OF INVENTION The present invention relates to a humanized or chimeric antibody binding to human CD3, compositions comprising said humanized or chimeric antibody, and use of said humanized or chimeric antibodies in treatment of a disease. BACKGROUND The Cluster of Differentiation 3 (CD3) has been known for many years and therefore has been subject of interest in many aspects. Specifically antibodies raised against CD3 or the T cell Receptor Complex, which CD3 is part of, are known. An in vitro characterization of recombinant chimeric CD3 isotype variants as well as a number of humanized OKT3 effector function variant antibodies has been described [1]. CD3 antibodies, e.g. muromonab-CD3, have been widely used in the treatment of acute allograft rejection. In addition, treatment with the anti-CD3 monoclonal antibody hOKT3 gamma1 (Ala-Ala) results in improved C-peptide responses and clinical parameters for at least 2 years after onset of type 1 diabetes in absence of continued immunosuppressive medications [2]. A promising approach to improve targeted antibody therapy is by delivering cytotoxic cells specifically to the antigen-expressing cancer cells. This concept of using T cells for efficient killing of tumor cells has been described [3]. However, initial clinical studies were rather disappointing mainly due to low efficacy, severe adverse effects (cytokine storm) and immunogenicity of the bispecific antibodies [4]. Advances in the design and application of bispecific antibodies have partially overcome the initial barrier of cytokine storm and improved clinical effectiveness without dose-limiting toxicities [5]. For example, certain bispecific antibodies targeting with one arm the antigen on the tumor cell and with the other arm for instance CD3 on T cells, and containing an active Fc fragment providing Fc receptor binding have been shown to induce tumor cell killing. Upon binding, a complex of T cells, tumor cells and effector cells that bind the antibody Fc region is potentially formed, leading to killing of the tumor cells [4]. Catumaxomab consists of a mouseIgG2a/ratIgG2b heavy chain heterodimer and has been found successful for the treatment of cancer-associated ascites after intraperitoneal application [6]. However, the mouse/rat hybrid is immunogenic [7] and cannot be applied for long-term treatment in humans. Frequent treatment-related adverse events attributed to catumaxomab included cytokine-release-related symptoms (i.e. pyrexia, nausea, vomiting, chills, tachycardia and hypotension) [8]-[9], which relate the potent polyclonal T cell activation by catumaxomab due to its active Fc fragment. Another antibody is ertumaxomab (HER2×CD3), which induces cytotoxicity in cell lines with HER2 expression. Ertumaxomab has been in Phase II clinical development for metastatic breast cancer [10]-[11]. Efficacy of CD3 bispecific antibodies and other CD3 bispecific antibody-based formats is dependent on several properties of bispecific antibodies, such as the affinity of the CD3 arm and/or the affinity to the target of the second arm and the target copy number on target cells. Some CD3 bispecific antibodies show high efficacy when the CD3 affinity is low (EpCamxCD3—Bortoletto 2002 PMID 12385030, MT103/Blinatumomab vs TandAb-Molhoj 2007 PMID 17083975), while other CD3 bispecifics demonstrate high efficacy using a high CD3 affinity (Reusch 2015, Mabs, PMID 25875246). In some cases high CD3 affinity is required, for example when arming ex vivo expanded activated T cells from patients with a bispecific antibody comprised of an anti-CD3 targeting arm and a second arm directed at a selected tumor-associated antigen. In the latter case, CD3 affinity should be high to retain the interaction with the expanded T cell when the product is infused back into the patient to mediate cytolysis of tumor cells (Reusch 2006 Clin Cancer Res PMID 16397041). However, high affinity antibodies to CD3, in contrast to low affinity ligands, are much less effective in TCR triggering at low copy number, since they display a stoichiometry of ˜1:1