WO2018215835A1 - Anti-cd47 x anti-mesothelin antibodies and methods of use thereof - Google Patents

Anti-cd47 x anti-mesothelin antibodies and methods of use thereof Download PDF

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Publication number
WO2018215835A1
WO2018215835A1 PCT/IB2018/000645 IB2018000645W WO2018215835A1 WO 2018215835 A1 WO2018215835 A1 WO 2018215835A1 IB 2018000645 W IB2018000645 W IB 2018000645W WO 2018215835 A1 WO2018215835 A1 WO 2018215835A1
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Prior art keywords
seq
acid sequence
amino acid
light chain
variable
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PCT/IB2018/000645
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English (en)
French (fr)
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Krzysztof Masternak
Nicolas Fischer
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Novimmune SA
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Novimmune SA
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Priority to AU2018272311A priority Critical patent/AU2018272311B2/en
Priority to JP2019565454A priority patent/JP7220161B2/ja
Priority to CN201880049297.7A priority patent/CN111201031B/zh
Priority to CA3065008A priority patent/CA3065008A1/en
Priority to EP18753224.7A priority patent/EP3630167A1/en
Publication of WO2018215835A1 publication Critical patent/WO2018215835A1/en
Anticipated expiration legal-status Critical
Priority to AU2025203336A priority patent/AU2025203336A1/en
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001102—Receptors, cell surface antigens or cell surface determinants
    • A61K39/001129—Molecules with a "CD" designation not provided for elsewhere
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001166—Adhesion molecules, e.g. NRCAM, EpCAM or cadherins
    • A61K39/001168—Mesothelin [MSLN]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/303—Liver or Pancreas
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins
    • C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies

Definitions

  • CD47 has also been implicated in cancer.
  • CD47 is overexpressed in various hematological and solid malignancies.
  • CD47 is a documented cancer stem cell/tumor initiating cell marker. It is thought that CD47 overexpression may help tumor cells to escape immune surveillance and killing by innate immune cells.
  • High levels of CD47 are also associated with poor clinical outcome in cancers such as, for example, leukemias, lymphomas, breast cancer, colon cancer, ovarian cancer, bladder cancer, prostate cancer, and/or glioma.
  • targeting CD47 would be useful in treating, delaying the progression of, or otherwise ameliorating a symptom of cancer.
  • the antibodies are considered to partially modulate, block, inhibit, reduce, antagonize, neutralize or otherwise interfere with the CD47-SIRPa interaction when the level of CD47-SIRPa interaction in the presence of the antibody is decreased by less than 95%, e.g. , 10%, 20%, 25%, 30%, 40%, 50%, 60%, 75%, 80%, 85% or 90% as compared to the level of CD47-SIRPa interaction in the absence of binding with an antibody described herein.
  • the bispecific antibody exhibits a "balanced" affinity for each of the two targets. In other embodiments, the bispecific antibody exhibits an "unbalanced" affinity for each of the two targets.
  • the first arm amino acid sequence includes a heavy chain comprising the amino acid sequence of SEQ ID NO: 2 and a light chain comprising the amino acid sequence of SEQ ID NO: 168
  • the second arm amino acid sequence includes a heavy chain amino comprising the acid sequence of SEQ ID NO: 2 and a light chain comprising the amino acid sequence of SEQ ID NO: 110.
  • Figure 5B Tumor Growth Inhibition (TGI).
  • Ke8H5 antibody includes a common variable heavy domain (SEQ ID NO: 1
  • SEQ ID NO: 114 encoded by the nucleic acid sequence shown in SEQ ID NO: 113 and includes a kappa variable light domain (SEQ ID NO: 152) encoded by the nucleic acid sequence shown in SEQ ID NO: 151.
  • the Ke8Gll antibody includes a common heavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappa light chain (SEQ ID NO: 46) encoded by the nucleic acid sequence shown in SEQ ID NO: 45.
  • the Ka3A3 antibody includes a common heavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappa light chain (SEQ ID NO: 62) encoded by the nucleic acid sequence shown in SEQ ID NO: 61.
  • the Kc4A4 antibody includes a common variable heavy domain (SEQ ID NO: 1]
  • the Kc4G9 antibody includes a common heavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambda light chain (SEQ ID NO: 86) encoded by the nucleic acid sequence shown in SEQ ID NO: 85.
  • the bispecific antibody Ka3 x 025 includes a common heavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQ ID NO: 1, a kappa light chain (SEQ ID NO: 56) encoded by the nucleic acid sequence shown in SEQ ID NO: 55, and a lambda light chain (SEQ ID NO: 98) encoded by the nucleic acid sequence shown in SEQ ID NO: 97.
  • SEQ ID NO: 2 common heavy chain
  • SEQ ID NO: 56 encoded by the nucleic acid sequence shown in SEQ ID NO: 55
  • SEQ ID NO: 98 a lambda light chain
  • control sequence refers to polynucleotide sequences which are necessary to effect the expression and processing of coding sequences to which they are ligated. The nature of such control sequences differs depending upon the host organism in prokaryotes, such control sequences generally include promoter, ribosomal binding site, and transcription termination sequence in eukaryotes, generally, such control sequences include promoters and transcription termination sequence.
  • control sequences is intended to include, at a minimum, all components whose presence is essential for expression and processing, and can also include additional components whose presence is advantageous, for example, leader sequences and fusion partner sequences.
  • the antibodies of the invention are monoclonal antibodies.
  • Monoclonal antibodies are generated, for example, by using the procedures set forth in the Examples provided herein.
  • Antibodies are also generated, e.g. , by immunizing BALB/c mice with combinations of cell transfectants expressing high levels of a given target on their surface. Hybridomas resulting from myeloma/B cell fusions are then screened for reactivity to the selected target.
  • the transfection step requires only small quantities of DNA and cells, typically 2 ⁇ g of plasmid DNA and 2x10 5 cells per well and the transfection carried out in a 6-well plate.
  • different mammalian cell lines can be used, in the examples given below, transformed human embryo kidney monolayer epithelial cells (PEAK cells) were transfected. These cells stably express the EBNA-1 gene, further supporting the episomal replication process, are semi-adherent and can be grown under standard conditions cell culture incubator (5% CC ; 37 °C in DMEM medium supplemented with 10% fetal calf serum). After 24h, cells were placed under selective conditions by adding medium containing 0.5-1 ⁇ g/mL puromycin, as cells harboring the episomal vector are resistant to this antibiotic.
  • PAK cells transformed human embryo kidney monolayer epithelial cells
  • the purification process was composed of three affinity steps. First, the CaptureSelectTM IgG-CHl affinity matrix (Thermo Fisher Scientific, Waltham, MA) was washed with PBS and then added in the clarified supernatant. After incubation overnight at +4°C, supernatants were centrifuged at 1000 g for 10 min, flow through was stored and resin washed twice with PBS. Then, the resin was transferred on spin columns and a solution containing 50 mM glycine at pH 2.7 was used for elution.
  • CaptureSelectTM IgG-CHl affinity matrix Thermo Fisher Scientific, Waltham, MA
  • the plasmid was then transfected into mammalian cells using a liposome based transfection reagent such as Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA).
  • the transfection step requires only small quantities of DNA and cells, typically 2xl0 5 cells and 2 ⁇ g of plasmid DNA per well and the transfection carried out in a 6-well plate.
  • different mammalian cell lines can be used, in the examples given below, transformed human embryo kidney monolayer epithelial cells (PEAK cells) were transfected.
  • Concentrations were determined using the dose response 5PL unweighted standard curve equation and an initial slope binding rate equation. Following 6-7 days culture period, the supernatant was harvested for IgG purification on FcXL affinity chromatography resin (Thermo Fisher Scientific, Waltham, MA) according to manufacturer's instructions. Briefly, supernatants from transfected cells were incubated overnight at +4°C with the resin. Samples were then centrifuged and the resin was transferred on a column filter for elution. The eluted IgG fraction was then desalted against PBS and the IgG content quantified by absorption at 280 nm. Purity and IgG integrity were verified by electrophoresis.
  • FIG. 1 shows strong binding of the MSLN monoclonal and bispecific antibodies to mesothelin expressed at the surface of mesothelin-transfected CHO cells. All MSLN antibodies show a high level of species cross-reactivity, as the MFI/antibody concentration curves obtained with human and cynomolgus MSLN-expressing CHO cells look very similar.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Oncology (AREA)
  • Cell Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
PCT/IB2018/000645 2017-05-26 2018-05-29 Anti-cd47 x anti-mesothelin antibodies and methods of use thereof Ceased WO2018215835A1 (en)

Priority Applications (6)

Application Number Priority Date Filing Date Title
AU2018272311A AU2018272311B2 (en) 2017-05-26 2018-05-29 Anti-CD47 x anti-mesothelin antibodies and methods of use thereof
JP2019565454A JP7220161B2 (ja) 2017-05-26 2018-05-29 抗ＣＤ４７ｘ抗メソテリン抗体およびそれを使用する方法
CN201880049297.7A CN111201031B (zh) 2017-08-25 2018-05-29 抗CD47 x抗间皮素抗体及其使用方法
CA3065008A CA3065008A1 (en) 2017-05-26 2018-05-29 Anti-cd47 x anti-mesothelin antibodies and methods of use thereof
EP18753224.7A EP3630167A1 (en) 2017-05-26 2018-05-29 Anti-cd47 x anti-mesothelin antibodies and methods of use thereof
AU2025203336A AU2025203336A1 (en) 2017-05-26 2025-05-08 Anti-CD47 x anti-mesothelin antibodies and methods of use thereof

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US201762511669P 2017-05-26 2017-05-26
US62/511,669 2017-05-26
US201762550387P 2017-08-25 2017-08-25
US62/550,387 2017-08-25

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EP (1) EP3630167A1 (enExample)
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CA (1) CA3065008A1 (enExample)
WO (1) WO2018215835A1 (enExample)

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WO2020114615A1 (en) * 2018-12-07 2020-06-11 Baxalta GmbH Bispecific antibodies binding factor ixa and factor x
WO2020115283A1 (en) * 2018-12-07 2020-06-11 Baxalta GmbH Bispecific antibodies binding factor ixa and factor x
EP3831849A1 (en) * 2019-12-02 2021-06-09 LamKap Bio beta AG Bispecific antibodies against ceacam5 and cd47
WO2022200389A1 (en) * 2021-03-22 2022-09-29 Novimmune S.A. Bispecific antibodies targeting cd47 and pd-l1 and methods of use thereof
JP2022551345A (ja) * 2019-12-17 2022-12-08 ファイザー・インク Ｃｄ４７、ｐｄ－ｌ１に特異的な抗体、およびその使用
WO2023006390A1 (en) * 2021-07-30 2023-02-02 Ludwig-Maximilians-Universität München Mesothelin antibodies and use thereof
WO2023006391A1 (en) * 2021-07-30 2023-02-02 Ludwig-Maximilians-Universität München Anti-cd47 antibodies and use thereof
WO2023139293A1 (en) 2022-01-24 2023-07-27 Novimmune Sa Composition and methods for the selective activation of cytokine signaling pathways
WO2023156625A1 (en) * 2022-02-18 2023-08-24 Adivo Gmbh Feline antibody library
WO2023170474A1 (en) * 2022-03-07 2023-09-14 Novimmune Sa Cd28 bispecific antibodies for targeted t cell activation
US12122850B2 (en) 2022-03-14 2024-10-22 LamKap Bio gamma AG Bispecific GPC3xCD28 and GPC3xCD3 antibodies and their combination for targeted killing of GPC3 positive malignant cells
WO2025114357A1 (en) * 2023-11-28 2025-06-05 Novimmune Sa Method of treating disease using anti-cd47 x anti-mesothelin antibodies as a sole agent and in combination with anti-pd-1 antibodies

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WO2017220555A1 (en) 2016-06-20 2017-12-28 F-Star Beta Limited Lag -3 binding members
GB201612520D0 (en) 2016-07-19 2016-08-31 F-Star Beta Ltd Binding molecules
ES3024433T3 (en) 2017-12-19 2025-06-04 F Star Therapeutics Ltd Fc binding fragments comprising a pd-l1 antigen-binding site
GB201811408D0 (en) 2018-07-12 2018-08-29 F Star Beta Ltd CD137 Binding Molecules
GB201811450D0 (en) 2018-07-12 2018-08-29 F Star Delta Ltd Mesothelin and CD137 binding molecules
CA3106046A1 (en) 2018-07-12 2020-01-16 F-Star Beta Limited Antibody molecules that bind pd-l1 and cd137
GB201811415D0 (en) 2018-07-12 2018-08-29 F Star Beta Ltd Anti-Mesothelin Anti bodies
GB201811403D0 (en) 2018-07-12 2018-08-29 F Star Beta Ltd Antibody molecules
ES3044118T3 (en) 2018-07-12 2025-11-26 Invox Pharma Ltd Antibody molecules that bind cd137 and ox40
GB201811410D0 (en) 2018-07-12 2018-08-29 F Star Beta Ltd OX40 Binding molecules
US20230391867A1 (en) * 2020-11-17 2023-12-07 Shenzhen Enduring Biotech, Ltd. Long acting bi-specific t cell engagers targeting cd3 and cd47
IL305828A (en) * 2021-03-22 2023-11-01 Novimmune Sa Bispecific antibodies targeting CD47 and PD-L1 and methods of using them

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