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JP-7856638-B2 - Improved anti-oxMIF antibody with reduced aggregation ability and reduced hydrophobicity

JP7856638B2JP 7856638 B2JP7856638 B2JP 7856638B2JP-7856638-B2

Inventors

  • シナグル，アレクサンデル
  • ミルキナ，イリナ
  • ケルスバウメル，ランドルフ
  • ティーレ，ロベルト・ミヒャエル

Assignees

  • オンコワン・リサーチ・アンド・ディベロップメント・ゲゼルシャフト・ミト・ベシュレンクテル・ハフツング

Dates

Publication Date
20260511
Application Date
20211001
Priority Date
20201002

Claims (20)

  1. A recombinant anti-oxMIF antibody or its antigen-binding fragment having reduced agglutination ability and reduced hydrophobicity, The following variable domains: (a) a light chain variable domain containing the sequence of Sequence ID No. 11 ; and Heavy chain variable domains containing the sequence of sequence number 6, 7, or 8; or (b) Light chain variable domain containing the sequence of Sequence ID No. 10; and A heavy chain variable domain containing the sequence of sequence number 7 or 8; or (c) Light chain variable domain containing the sequence of Sequence ID No. 13; and A heavy chain variable domain containing the sequence of Sequence ID No. 8; or ( d ) Light chain variable domain containing the sequence of Sequence ID No. 9; and Heavy chain variable domain containing the sequence of sequence number 8 , Includes, A recombinant anti-oxMIF antibody or its antigen-binding fragment, which exhibits reduced agglutination ability and hydrophobicity compared to an antibody or its antigen-binding fragment containing the sequences of SEQ ID NO: 6 and SEQ ID NO: 9.
  2. (a) The sequence of SEQ ID NO: 2 or a heavy chain constant region having at least 95% sequence identity with the sequence of SEQ ID NO: 2, and containing amino acids 239D and 332E; or (b) The sequence of SEQ ID NO: 3 or a heavy chain constant region having at least 95% sequence identity with the sequence of SEQ ID NO: 3, and containing amino acids 239D, 268F, 324T and 332E; or (c) The sequence of SEQ ID NO: 4 or a heavy chain constant region having at least 95% sequence identity with the sequence of SEQ ID NO: 4, and containing amino acids 239D and 332E; or (d) The sequence of SEQ ID NO: 5 or a heavy chain constant region having at least 95% sequence identity with the sequence of SEQ ID NO: 5, and containing amino acids 235V, 243L, 292P, 300L and 396L, and including a heavy chain constant region, The amino acid positions are numbered according to the EU numbering index. The recombinant anti-oxMIF antibody according to claim 1, having enhanced effector function.
  3. The anti-oxMIF antibody according to claim 1, comprising any one sequence of SEQ ID NOs: 6, 11, and SEQ ID NOs: 1, 2, 3, 4, or 5.
  4. The anti-oxMIF antibody according to claim 1, comprising any one sequence of SEQ ID NOs: 7, 10, and SEQ ID NOs: 1, 2, 3, 4, or 5.
  5. The anti-oxMIF antibody according to claim 1, comprising any one sequence of SEQ ID NOs: 7, 11, and SEQ ID NOs: 1, 2, 3, 4, or 5.
  6. The anti-oxMIF antibody according to claim 1, comprising any one sequence of SEQ ID NOs: 8, 11, and SEQ ID NOs: 1, 2, 3, 4, or 5.
  7. The anti-oxMIF antibody according to claim 1, comprising any one sequence of SEQ ID NOs: 8, 13, and SEQ ID NOs: 1, 2, 3, 4, or 5.
  8. The anti-oxMIF antibody according to claim 1, comprising any one sequence of SEQ ID NOs: 8, 10, and SEQ ID NOs: 1, 2, 3, 4, or 5.
  9. The anti-oxMIF antibody according to claim 1, comprising the sequences of SEQ ID NOs: 8, 11, 4, and 38, and optionally the sequence of SEQ ID NOs: 1, 2, 3, 4, or 5.
  10. The anti-oxMIF antibody according to claim 1, comprising the sequences of sequence numbers 8, 13, 4, and 38, and optionally sequence number 1, 2, 3, 4, or 5.
  11. An anti-oxMIF antibody according to any one of claims 1 to 10, selected from the group consisting of bispecific antibodies, scFv, (scFv)2, scFvFc, Fab, Fab' , F (ab')2, Fab'-SH, Fab-scFv fusion, Fab-(scFv)2-fusion, Fab-scFv-Fc, Fab-(scFv) 2 -Fc, a fusion protein of two different types of single-chain antibodies (BiTE), and a minibody.
  12. An antibody according to any one of claims 1 to 11 , for use in the preparation of pharmaceuticals.
  13. A pharmaceutical composition comprising the antibody described in any one of claims 1 to 11 , optionally together with a pharmaceutical carrier or adjuvant.
  14. The pharmaceutical composition according to claim 13 , comprising an antibody according to any one of claims 1 to 11 in an amount of 10 to 250 mg/ml.
  15. The pharmaceutical composition according to claim 14, comprising an antibody according to any one of claims 1 to 11 in a concentration of 50 mg/ml .
  16. A pharmaceutical composition according to any one of claims 13 to 15, formulated for subcutaneous administration.
  17. The pharmaceutical composition according to any one of claims 13 to 16, for administration as a single substance or, preferably, in combination with a further pharmaceutical composition comprising one or more active substances selected from the group consisting of antiviral substances, anti-inflammatory substances, anticancer substances, anti - angiogenic substances, and antibiotics.
  18. A pharmaceutical composition according to any one of claims 13 to 17, for use in the treatment of patients suffering from inflammatory diseases, hyperproliferative disorders, infectious diseases, or cancer.
  19. The pharmaceutical composition according to claim 18 for use in the treatment of asthma, vasculitis, arthritis, sepsis, septic shock, endotoxin shock, toxic shock syndrome, acquired respiratory distress syndrome, glomerulonephritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, peritonitis, nephritis and psoriasis, colorectal cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, or lung cancer.
  20. An isolated nucleic acid encoding an antibody according to any one of claims 1 to 11 .

Description

This invention relates to anti-oxMIF antibodies having improved properties, such as reduced aggregation ability and reduced hydrophobicity due to selected amino acid substitutions in the light chain and heavy chain variable domains, and optionally, increased effector function due to further substitutions in the heavy chain constant region, as well as their use in the treatment of oxMIF-related diseases. The cytokine macrophage migration inhibitory factor (MIF) was described as early as 1966 (David, J.R., 1966; Bloom B.R. and Bennett, B., 1966). However, MIF differs significantly from other cytokines and chemokines because it is constitutively expressed, stored in the cytoplasm, and present in the circulation of healthy subjects. Due to the ubiquity of this protein, MIF may be considered an unsuitable target for therapeutic intervention. However, MIF exists in two immunologically distinct isoforms called reduced MIF (redMIF) and oxidized MIF (oxMIF) (Thiele M. et al., 2015). RedMIF has been found to be a highly expressed isoform of MIF that can be found in the cytoplasm and in the circulation of any subject. redMIF appears to represent a latent, inactive storage form (Schinagl. A. et al., 2018). In contrast, oxMIF can be detected in tumor tissue, specifically in tumor tissue from patients with colorectal cancer, pancreatic cancer, ovarian cancer, and lung cancer, which exhibit high tumor specificity for oxMIF (Schinagl. A. et al., 2016). However, it also appears to be a physiologically related disease-associated isoform that can be detected in the circulation of patients with inflammatory diseases (Thiele et al., 2015). The number of successful drug targets for treating cancers like the oxMIF-positive cancer described above is limited. For example, while over 300 potential immuno-oncological targets have been listed, many clinical studies focus on anti-PD1 and anti-PDL1 antibodies (Tang J., et al. 2018). Therefore, the scientific and medical communities are keen to discover tumor-specific antigen-targeted drugs to increase treatment options for cancer patients with poor prognoses. A monoclonal anti-MIF antibody is described in International Publication No. 2009/086920 A1. Antibodies targeting oxMIF demonstrated efficacy in in vitro and in vivo models of inflammation and cancer (Hussain F. et al., 2013; Schinagl. A. et al., 2016; Thiele et al., 2015). The oxMIF-specific antibody (imarumab) demonstrated an acceptable safety profile, excellent tissue penetration, and indications for antitumor activity in Phase 1 clinical trials (Mahalingam D. et al., 2015; Mahalingam D. et al., 2020). A bispecific anti-oxMIF/anti-CD3 antibody is disclosed in International Publication No. 2019/234241 A1. Protein aggregation, specifically antibody aggregation, is frequently observed at several stages of bioprocesses, including protein expression, purification, and storage. Antibody aggregation can affect the overall yield of therapeutic protein production processes and may contribute to the stability and immunogenicity of therapeutic antibodies. Therefore, antibody protein aggregation remains a significant problem in their occurrence and a major concern in antibody production. Antibody aggregation can be induced by partial unfolding of its domains after monomer-monomer association, leading to nucleation and aggregate proliferation. While the aggregation tendencies of antibodies and antibody-based proteins can be influenced by external experimental conditions, they are strongly dependent on their potential antibody properties, which are determined by their sequence and structure. For example, resistance to aggregation can be achieved by stabilizing the native state (i.e., resistance to unfolding) or by reducing the unfolded or partially folded properties of the protein towards aggregation. A drawback of stabilizing the native state is that the protein may be exposed to environments that cause unfolding. Generally, when a protein denatures or unfolds, amino acid residues that mediate intramolecular contacts within the protein are exposed. Such exposure often makes the protein more susceptible to intermolecular contact and aggregation. In contrast to unfolding-resistant proteins, proteins with reduced aggregation properties when unfolded simply refold to a physiologically active non-aggregated state after exposure to such environments. The agglutination resistance or tendency of antibodies and proteins containing their antigen-binding domains is typically limited by the numerous agglutination-prone domains (maybe multiple domains) present and the strength of their interactions with (if present) surrounding domains. This is because, once the domain unfolds, if it cannot refold, it can interact with other domains within the same protein or other proteins, potentially forming aggregates. The constant domains of antibodies generally do not aggregate and do not change significantly. Therefore, the weakest domains of antibodies in terms o