WO2024074145A1 - Anticorps bispécifique se liant à baffr et cd3 et utilisation associée - Google Patents

Anticorps bispécifique se liant à baffr et cd3 et utilisation associée Download PDF

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WO2024074145A1
WO2024074145A1 PCT/CN2023/123288 CN2023123288W WO2024074145A1 WO 2024074145 A1 WO2024074145 A1 WO 2024074145A1 CN 2023123288 W CN2023123288 W CN 2023123288W WO 2024074145 A1 WO2024074145 A1 WO 2024074145A1
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seq
antigen binding
bispecific antibody
cancer
sequence
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PCT/CN2023/123288
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English (en)
Chinese (zh)
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周冲
殷刘松
姜晓玲
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盛禾(中国)生物制药有限公司
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Publication of WO2024074145A1 publication Critical patent/WO2024074145A1/fr

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • 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
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins

Definitions

  • the present invention belongs to the field of biomedicine, and specifically relates to a bispecific antibody combining BAFFR and CD3 and an application thereof.
  • CD3 T cell surface glycoprotein CD3, signal transduction co-receptor of T cell receptor, which contains subunits ⁇ , ⁇ , ⁇ and ⁇
  • CD3 is a differentiation antigen expressed on the surface of all T lymphocytes, which mainly mediates the transduction of T cell activation signals. It plays an important role in the anti-infection immunity of the body's immune system.
  • the CD3 molecule forms a stable TCR-CD3 complex with the T cell antigen receptor.
  • the extracellular region recognizes and binds to the major histocompatibility complex class II molecules, enhancing the stability of the binding between the T cell antigen receptor and the MHC molecule; the intracellular region enhances the activation signal transduced by leukocyte CD3, thereby participating in and regulating the activation of the immune system.
  • the number of CD3-positive lymphocytes is an important indicator for measuring the cellular immunity of the body.
  • BAFFR The only ligand of BAFFR is BAFF.
  • BAFFR and BAFF work together to activate the NF-kB signaling pathway in B cells, promoting the proliferation and activation of B cells.
  • the BAFF-BAFFR signaling pathway and the BCR pathway are key pathways for the maturation and activation of Immature B cells. Blocking the dual pathways can completely eliminate peripheral B lymphocytes and inhibit the development and proliferation of B lymphocytes.
  • BAFFR-targeted antibody drugs C55 and C90 are as effective as Rituximab, and can completely eliminate lymphoma cells in the blood of mice and prolong the tumor-free survival of mice.
  • C90 can still completely eliminate lymphoma cells in the mouse blood and extend the tumor-free survival of mice to 100 days (rituximab survival time is within 20 days).
  • BAFFR-targeted antibodies can effectively solve the problems of ineffectiveness and recurrent drug resistance of ibrutinib.
  • the anti-tumor activity of the ADCC activity of monoclonal antibody drugs is limited.
  • T cells have been proven to be an effective strategy as effective anti-tumor cells. Therefore, the development of T cell engager therapy that can target both T cell receptors and tumor-associated antigens (TAA) is widely used.
  • TAA tumor-associated antigens
  • Anti-CD3 and anti-BAFFR bispecific antibodies can activate endogenous T cells by bispecifically binding to BAFFR on the surface of tumor cells and CD3 on the surface of T cells, leading to the directed lysis of BAFFR-positive tumor cells, thereby achieving the purpose of treating tumors.
  • the inventors have developed a bispecific antibody with good performance that can bind to BAFFR and CD3.
  • the bispecific antibody of the present invention maximizes the tumor killing effect through multivalent binding of tumor-associated antigens while controlling the toxicity of CD3: BAFFR antibody multivalently binds to BAFFR-positive tumors, and the CD3 antibody at the other end binds to T cells.
  • the bispecific antibody acts as a connector to bring T cells and tumor cells closer to form an immune synapse, allowing T cells to kill tumors; the monovalent CD3 antibody reduces toxicity and can Better form immune synapses to achieve more effective, specific and safer anti-tumor responses.
  • the present invention provides a bispecific antibody, which comprises: (a) a first antigen-binding portion that specifically binds to a first antigen, wherein the first antigen is BAFFR; (b) a second antigen-binding portion that specifically binds to a second antigen, wherein the second antigen is BAFFR; and (c) a third antigen-binding portion that specifically binds to a third antigen, wherein the third antigen is CD3.
  • the first antigen binding moiety is a full length antibody consisting of two heavy chains and two light chains.
  • the second antigen binding moiety is an antibody fragment comprising a heavy chain variable domain (VH) and/or a light chain variable domain (VL).
  • the third antigen binding moiety is an antibody fragment comprising a heavy chain variable domain (VH) and/or a light chain variable domain (VL).
  • the second antigen binding moiety is Fab, Fab', scFab, F(ab')2, Fv, dsFv or scFv.
  • the second antigen binding moiety is Fab.
  • the third antigen binding moiety is Fab, Fab', scFab, F(ab')2, Fv, dsFv or scFv.
  • the third antigen binding moiety is scFv.
  • the second antigen binding moiety is fused to the N-terminus of one heavy chain of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the C-terminus of one heavy chain of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-terminus and C-terminus of one heavy chain of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-termini of the two heavy chains of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the C-termini of the two heavy chains of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-terminus and C-terminus of the two heavy chains of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-terminus of one light chain of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the C-terminus of one light chain of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-terminus and C-terminus of one light chain of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-termini of the two light chains of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the C-termini of the two light chains of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-terminus and C-terminus of the two light chains of the first antigen binding moiety.
  • the third antigen binding moiety replaces one or two Fab regions of the first antigen binding moiety.
  • the third antigen binding moiety replaces the Fab region of the first antigen binding moiety fused to the second antigen binding moiety.
  • the third antigen binding moiety replaces one or two Fv regions of the first antigen binding moiety.
  • the third antigen binding moiety replaces the Fv region fused to the second antigen binding moiety.
  • the Fv region of the first antigen binding moiety is not limited to one or two Fv regions of the first antigen binding moiety.
  • the second antigen binding moiety is fused to the N-terminus of one heavy chain of the first antigen binding moiety and the third antigen binding moiety replaces the Fab region of the first antigen binding moiety fused to the second antigen binding moiety.
  • the second antigen binding moiety is fused to the N-terminus of one heavy chain of the first antigen binding moiety and the third antigen binding moiety replaces the Fv region of the first antigen binding moiety fused to the second antigen binding moiety.
  • the bispecific antibody comprises a first Fc region and a second Fc region.
  • the first Fc region and the second Fc region are the same or different.
  • the Fc region is selected from IgG, IgA, IgD, IgE, IgM and variants thereof.
  • the Fc region is selected from IgG1, IgG2, IgG3, IgG4 and variants thereof.
  • the Fc region comprises one or more amino acid mutations, preferably amino acid substitutions, insertions or deletions.
  • the first Fc region is knob-Fc and the second Fc region is hole-Fc.
  • the first Fc region is hole-Fc and the second Fc region is knob-Fc.
  • the VH and VL of the third antigen binding moiety are swapped.
  • the second antigen binding moiety is fused to the N-terminus of one heavy chain of the first antigen binding moiety
  • the third antigen binding moiety replaces the Fab region of the first antigen binding moiety fused to the second antigen binding moiety
  • the VH and VL of the third antigen binding moiety are interchanged.
  • the second antigen binding moiety is fused to the N-terminus of one heavy chain of the first antigen binding moiety
  • the third antigen binding moiety replaces the Fv region of the first antigen binding moiety fused to the second antigen binding moiety
  • the VH and VL of the third antigen binding moiety are interchanged.
  • the second and third antigen binding moieties are fused to the first antigen binding moiety via a linker.
  • the linker is a peptide linker.
  • the peptide linker is a GS linker or a mutant human IgG hinge.
  • the peptide linker has an amino acid sequence as shown in (G4S) x , where x is an integer selected from 1-6; preferably, the peptide linker is (G4S) 2 , (G4S) 3 or (G4S) 4. More preferably, the peptide linker is (G4S) 2 .
  • the first antigen binding portion specifically binds to BAFFR, wherein HCDR1 of the first antigen binding portion is as shown in SEQ ID NO:3, or a sequence having at least 80% identity with SEQ ID NO:3; HCDR2 is as shown in SEQ ID NO:4, or a sequence having at least 80% identity with SEQ ID NO:4; HCDR3 is as shown in SEQ ID NO:5, or a sequence having at least 80% identity with SEQ ID NO:5; LCDR1 is as shown in SEQ ID NO:6, or a sequence having at least 80% identity with SEQ ID NO:6; LCDR2 is as shown in SEQ ID NO:7, or a sequence having at least 80% identity with SEQ ID NO:7; LCDR3 is as shown in SEQ ID NO:8, or a sequence having at least 80% identity with SEQ ID NO:8.
  • the second antigen binding portion specifically binds BAFFR, wherein HCDR1 of the second antigen binding portion is as shown in SEQ ID NO:3, or a sequence having at least 80% identity with SEQ ID NO:3; HCDR2 is as shown in SEQ ID NO:4, or a sequence having at least 80% identity with SEQ ID NO:4; HCDR3 is as shown in SEQ ID NO:5, or a sequence having at least 80% identity with SEQ ID NO:5; LCDR1 is as shown in SEQ ID NO:6, or a sequence having at least 80% identity with SEQ ID NO:6; LCDR2 is as shown in SEQ ID NO:7, or a sequence having at least 80% identity with SEQ ID NO:7; LCDR3 is as shown in SEQ ID NO:8, or a sequence having at least 80% identity with SEQ ID NO:8.
  • the third antigen binding portion specifically binds to CD3, wherein HCDR1 of the third antigen binding portion is as shown in SEQ ID NO:11, or a sequence having at least 80% identity with SEQ ID NO:11; HCDR2 is as shown in SEQ ID NO:12, or a sequence having at least 80% identity with SEQ ID NO:12; HCDR3 is as shown in SEQ ID NO:13, or a sequence having at least 80% identity with SEQ ID NO:13; and LCDR1 is as shown in SEQ ID NO:14, or a sequence having at least 80% identity with SEQ ID NO:14; LCDR2 is as shown in SEQ ID NO:15, or a sequence having at least 80% identity with SEQ ID NO:15; LCDR3 is as shown in SEQ ID NO:16, or a sequence having at least 80% identity with SEQ ID NO:16.
  • HCDR1 of the third antigen binding portion is as shown in SEQ ID NO:11, or a sequence having at least 80% identity with S
  • the first antigen binding portion specifically binds to BAFFR, wherein the heavy chain variable region VH of the first antigen binding portion is as shown in SEQ ID NO:1, or a sequence with at least 80% identity to SEQ ID NO:1; the light chain variable region VL is as shown in SEQ ID NO:2, or a sequence with at least 80% identity to SEQ ID NO:2.
  • the second antigen binding portion specifically binds to BAFFR, wherein the heavy chain variable region VH of the second antigen binding portion is as shown in SEQ ID NO:1, or a sequence having at least 80% identity with SEQ ID NO:1; the light chain variable region VL is as shown in SEQ ID NO:2, or a sequence having at least 80% identity with SEQ ID NO:2.
  • the third antigen binding portion specifically binds to CD3, wherein the heavy chain variable region VH of the third antigen binding portion is as shown in SEQ ID NO:9, or a sequence with at least 80% identity with SEQ ID NO:9; the light chain variable region VL is as shown in SEQ ID NO:10, or a sequence with at least 80% identity with SEQ ID NO:10.
  • the present invention also provides an isolated nucleic acid molecule comprising a nucleotide sequence encoding any of the above-mentioned bispecific antibodies.
  • the cancer is selected from human brain astrocytoma, human pharyngeal cancer, adrenal tumor, AIDS-related cancer, alveolar soft tissue sarcoma, astrocytoma, bladder cancer, bone cancer, brain and spinal cord cancer, metastatic brain tumor, breast cancer, carotid body tumor, cervical cancer, chondrosarcoma, chordoma, renal chromophobe cell carcinoma, clear cell carcinoma, colon cancer, colorectal cancer, desmoplastic small round cell tumor, ependymoma, Ewing tumor, extraskeletal myxoid chondrosarcoma, fibrous dysplasia, fibrous dysplasia, gallbladder or bile duct cancer, gastric cancer, gestational trophoblastic disease, germ cell tumor, head and neck cancer, hepatocellular carcinoma, islet cell tumor, Kaposi's
  • the present invention also provides the use of any of the above-described bispecific antibodies in the preparation of a medicament for treating an autoimmune disease.
  • the autoimmune disease is selected from graft-versus-host disease, rheumatoid arthritis, Crohn's disease, multiple sclerosis, colitis, psoriasis, autoimmune uveitis, pemphigus, epidermolysis bullosa, or type I diabetes.
  • the use is achieved by one or more of tumor immunotherapy, cell therapy, or gene therapy.
  • the present invention also provides a pharmaceutical composition, comprising any of the bispecific antibodies described above and a pharmaceutically acceptable carrier, diluent or excipient.
  • the present invention also provides an antibody-drug conjugate, comprising any of the above-mentioned bispecific antibodies.
  • the conjugated drug is selected from cytotoxins, small molecule chemical drugs or immunotoxins.
  • VH antibody heavy chain variable region
  • VL antibody light chain variable region
  • CDR complementarity determining region in an immunoglobulin variable region
  • IgG immunoglobulin G.
  • antibody refers to a natural immunoglobulin or an immunoglobulin prepared by partial or complete synthesis. Antibodies can be reconstructed and separated from natural resources such as plasma or serum in which the antibody is naturally present, or from the culture supernatant of hybridoma cells that produce the antibody, from animal immune serum, or from phage library screening. Alternatively, it can be partially or completely synthesized using techniques such as gene recombination. Preferred antibodies include, for example, antibodies of isotypes of immunoglobulins or subclasses of these isotypes.
  • Known human immunoglobulins include 9 categories (isotypes) of IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD, IgE, and IgM.
  • antibodies of the present invention can include IgG1, IgG2, IgG3, and/or IgG4.
  • the partial antibodies used herein are immunoglobulin molecules consisting of two pairs of polypeptide chains, each pair having one light chain (LC) and one heavy chain (HC).
  • Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH).
  • the heavy chain constant region consists of three domains (CH1, CH2, and CH3).
  • Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL), or only a light chain constant region (CL).
  • the light chain constant region consists of one domain CL.
  • the constant domain does not directly participate in the binding of the antibody to the antigen, but exhibits a variety of effector functions, such as mediating the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.
  • the VH and VL regions can also be subdivided into regions with high variability (called complementarity determining regions (CDRs)), interspersed with more conservative regions called framework regions (FRs).
  • CDRs complementarity determining regions
  • FRs framework regions
  • Each VH and VL consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • the variable regions (VH and VL) of each heavy chain/light chain pair form an antigen binding site, respectively.
  • antigen binding portion or “antigen binding fragment” refers to one or more portions of an antibody that retain the ability to bind to the antigen to which the antibody binds.
  • antigen binding fragments include (1) Fab fragments; (2) F(ab')2 fragments; (3) Fd fragments, consisting of VH and CH1 domains; (4) Fv fragments; (5) dAb fragments, consisting of VH domains; (6) CDRs, separated complementarity determining regions.
  • the two domains VL and VH of the Fv fragment are encoded by separate genes, recombinant methods can be used to connect them through synthetic linkers so that they can be produced as a single protein chain (called single-chain Fv (scFv)) in which the VL and VH regions are paired to form a monovalent molecule.
  • single-chain antibodies are also intended to be included in the "antigen-binding fragment" of the term antibody.
  • Such antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for functionality in the same manner as for intact antibodies.
  • Antigen-binding portions can be produced by recombinant DNA technology or by enzymatic or chemical cleavage of intact immunoglobulins.
  • Antigen-binding fragments can also be incorporated into single-chain molecules comprising a pair of tandem Fv fragments (VH-CH1-VH-CH1), which form a pair of antigen-binding regions together with complementary light chain polypeptides.
  • Fab fragment consists of a complete L chain and the variable region domain (VH) of the H chain and the first constant domain (CH1) of one heavy chain.
  • VH variable region domain
  • CH1 first constant domain
  • Fab fragments can be produced recombinantly or by papain digestion of full-length antibodies.
  • Fab' fragment differs from the Fab fragment in that a few additional residues are added at the carboxyl terminus of the CH1 domain, including
  • Fab' can be produced by treating F(ab')2 that specifically recognizes and binds to an antigen with a reducing agent such as dithiothreitol.
  • F(ab')2 fragment was originally produced as a pair of Fab' fragments with a hinge cysteine between them.
  • F(ab')2 fragments can be produced recombinantly or by pepsin digestion of intact antibodies (which removes most of the Fc region while leaving part of the hinge region intact).
  • F(ab')2 fragments can be dissociated (into two Fab' molecules) by treatment with a reducing agent such as ⁇ -mercaptoethanol.
  • scFab refers to a single-chain Fab fragment, in which a polypeptide linker is introduced between the heavy chain variable domain (VH) and the light chain (CL) to form a single-chain Fab fragment (scFab).
  • Fv is the smallest antibody fragment containing a complete antigen recognition and binding site.
  • the fragment is composed of a dimer formed by a heavy chain variable region domain and a light chain variable region domain through tight non-covalent binding.
  • the folding of these two domains produces six hypervariable loops (3 loops from the H chain and 3 loops from the L chain), which contribute to the amino acid residues for antigen binding and give the antibody antigen binding specificity.
  • six hypervariable loops (3 loops from the H chain and 3 loops from the L chain), which contribute to the amino acid residues for antigen binding and give the antibody antigen binding specificity.
  • a single variable domain has the ability to recognize and bind antigens, its affinity is lower than that of the complete binding site.
  • the term "scFv" fragment refers to an antibody fragment comprising the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain.
  • the Fv polypeptide may further comprise a polypeptide linker between the VH and VL domains, which enables the scFv to form a desired structure for antigen binding.
  • the "scFv-Fc” fragment comprises an scFv connected to an Fc domain.
  • the Fc domain can be connected to the C-terminus of the scFv.
  • the Fc domain may be after VH or VL.
  • the Fc domain may be any suitable Fc domain known in the art or described herein. In some cases, the Fc domain is an IgG1 Fc domain.
  • dsFv refers to a disulfide-stabilized Fv fragment.
  • dsFv a polypeptide in which one amino acid residue in each VH and VL is replaced by a cysteine residue is connected via a disulfide bond between the cysteine residues.
  • each amino acid in the framework region of VH and VL is mutated to cysteine, which in turn forms a stable interchain disulfide bond.
  • position 44 in VH and position 100 in VL are mutated to cysteine.
  • dsFv encompasses both dsFv (molecules in which VH and VL are connected by interchain disulfide bonds rather than linker peptides) or scdsFv (molecules in which VH and VL are connected by linkers and interchain disulfide bonds) known in the art.
  • epitope refers to an antigenic determinant in an antigen, and refers to an antigenic site bound by a domain of an antigen binding molecule comprising an antibody variable region disclosed in this specification. Therefore, an epitope can be defined based on its structure. In addition, the epitope can also be defined based on the antigen binding activity in an antigen binding molecule that recognizes the epitope. When the antigen is a peptide or polypeptide, the epitope can be specified by the amino acid residues that form the epitope; when the epitope is a sugar chain, the epitope can be determined by its specific sugar chain structure.
  • the term "specificity" means that one of the molecules involved in specific binding does not show any significant binding to molecules other than one or more of the binding partner molecules.
  • the term is also used when the domain containing the antibody variable region is specific for a particular epitope among multiple epitopes in an antigen.
  • the antigen-binding molecule containing the domain containing the antibody variable region can bind to various antigens having the epitope.
  • bispecific antibody refers to a protein molecule that can specifically bind to two target antigens or target antigen epitopes.
  • bispecific antigen-binding protein comprising an antibody or antigen-binding fragment (e.g., Fab, scFv, etc.) and "bispecific antibody” and “bi-antibody” can be used interchangeably.
  • knock-Fc refers to replacing an amino acid residue in the CH3 domain of the first subunit of the Fc domain with an amino acid residue having a larger side chain volume, thereby generating a protrusion in the CH3 domain of the first subunit that can be positioned in the recess in the CH3 domain of the second subunit. For example, by mutating serine T at position 366 of CH3 of a heavy chain to tryptophan W, a protruding "knob"-like protrusion is formed.
  • hole-Fc refers to replacing the amino acid residues in the CH3 domain of the second subunit of the Fc domain with amino acid residues having a smaller side chain volume, thereby generating a depression in the CH3 domain of the second subunit in which the protrusion in the CH3 domain of the first subunit can be positioned.
  • hole-Fc refers to replacing the amino acid residues in the CH3 domain of the second subunit of the Fc domain with amino acid residues having a smaller side chain volume, thereby generating a depression in the CH3 domain of the second subunit in which the protrusion in the CH3 domain of the first subunit can be positioned.
  • fusion refers to the connection of two amino acid sequences into a new sequence through a linker or other technical means, thereby forming a new artificial protein or antibody.
  • linker or "L1" used to connect two protein domains refers to a connecting polypeptide sequence that is used to connect protein domains and has a certain degree of flexibility. The use of the linker will not cause the loss of the original function of the protein domain.
  • variable region or “variable domain” of an antibody refers to the variable region (VL) of an antibody light chain or the variable region (VH) of an antibody heavy chain, either alone or in combination.
  • VL variable region
  • VH variable region
  • the variable regions of the heavy and light chains are each composed of four framework regions (FRs) connected by three complementary determining regions (CDRs) (also referred to as hypervariable regions).
  • FRs framework regions
  • CDRs complementary determining regions
  • the CDRs in each chain are held together closely by the FRs and contribute to the formation of the antigen-binding site of the antibody together with the CDRs from the other chain.
  • variable refers to the fact that certain segments of the variable domain are widely different in sequence between antibodies.
  • the V domain mediates antigen binding and defines the specificity of a particular antibody for its specific antigen.
  • variability is not evenly distributed over the entire variable domain range. Instead, it is concentrated in three segments called hypervariable regions (HVRs) within the light chain and heavy chain variable domains.
  • HVRs hypervariable regions
  • FRs framework regions
  • the variable domains of the native heavy and light chains each contain four FR regions, most of which adopt a ⁇ -folded configuration, connected by three HVRs, which form a loop connection, and in some cases form a part of the ⁇ -folded structure.
  • the HVR in each chain is closely held together by the FR region, and together with the HVRs of other chains, contribute to the formation of the antigen binding site of the antibody.
  • the constant domain is not directly involved in the combination of the antibody and the antigen, but exhibits various effector functions, such as participating in the antibody-dependent cellular toxicity of the antibody.
  • antibody drug conjugate refers to a binding protein (such as an antibody or an antigen-binding fragment thereof) connected to one or more conjugated drugs (which may optionally be a therapeutic agent or a cytotoxic agent), and its structure generally consists of three parts: an antibody or antibody-like ligand, a drug portion, and a linker that couples the antibody or antibody-like ligand and the drug.
  • ADCs generally have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 drugs conjugated to the antibody.
  • polypeptide refers to an amino acid chain of any length, regardless of modification (e.g., phosphorylation or glycosylation).
  • the term polypeptide includes proteins and fragments thereof.
  • Polypeptides can be "exogenous,” meaning that they are “heterologous,” i.e., foreign to the host cell utilized, such as human polypeptides produced by bacterial cells.
  • Polypeptides are disclosed herein as sequences of amino acid residues. Those sequences are written from left to right in the direction of the amino terminus to the carboxyl terminus. Amino acid residue sequences are named by three-letter or one-letter codes according to standard nomenclature.
  • amino acid refers to the twenty common naturally occurring amino acids.
  • amino acid also includes non-natural amino acids. Any suitable non-natural amino acid can be used.
  • the non-natural amino acid comprises a reactive portion for conjugating the agent to MIAC.
  • identity is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to obtain the maximum percentage sequence identity. Comparisons for purposes of determining percentage amino acid sequence identity can be performed in a variety of ways within the skill of the art, for example using publicly available computer software such as BLAST software or FASTA packages.
  • the term "at least 80% identity” refers to the amino acid residues in the candidate sequence that are identical to those in the reference polypeptide sequence.
  • the percentage of identical amino acid residues is above 80%, including 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and 100%.
  • amino acid mutation refers to the presence of amino acid mutations or changes in variant proteins or polypeptides compared to the original protein or polypeptide, including the insertion, deletion or substitution of one or more amino acids in the original protein or polypeptide.
  • nucleic acid molecule refers to DNA molecules and RNA molecules. Nucleic acid molecules can be single-stranded or double-stranded, but are preferably double-stranded DNA. A nucleic acid is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence.
  • host cell refers to a cell that has been or can be transformed with a nucleic acid sequence and thereby expresses a selected target gene.
  • the term includes the offspring of a parent cell, whether or not the offspring is identical to the original parent cell in morphology or genetic composition, as long as the offspring has the selected target gene.
  • Commonly used host cells include bacteria, yeast, mammalian cells, etc.
  • vector refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked.
  • the term includes vectors that are self-replicating nucleic acid structures as well as vectors that are incorporated into the genome of a host cell into which they are introduced.
  • Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked and are referred to herein as "expression vectors.”
  • pharmaceutically acceptable carrier includes any of the standard pharmaceutical carriers, such as phosphate-buffered saline solutions, water, and emulsions, as well as various types of wetting agents.
  • positive control refers to a natural or engineered cell or antibody that can bind to or express a target protein.
  • the positive control referred to herein refers to a single-target positive control.
  • control refers to the use of the same species, subtype, dose, immunoglobulin and subtype of immunoglobulin, and the same marker as the experimental sample in the same experiment to eliminate the experimental background effect of non-specific binding samples on the experimental values, as a control to better illustrate the experimental effect.
  • affinity refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antigen-binding module of MIAC) and its binding partner (e.g., an antigen). Within each antigenic site, the variable region of the antibody “arm” interacts with the antigen at multiple amino acid sites through weak non-covalent forces; the greater the interaction, the stronger the affinity.
  • binding affinity refers to the intrinsic binding affinity of a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen).
  • the affinity of a molecule X for its partner Y can generally be represented by a dissociation constant (Kd). Affinity can be measured by common methods known in the art, such as by using surface plasmon resonance (SPR) technology (e.g., an instrument) or biolayer interferometry (e.g., an instrument) to measure.
  • SPR surface plasmon resonance
  • biolayer interferometry e.g., an instrument
  • effector cell is a leukocyte that expresses one or more FcRs and performs effector functions.
  • the effector cell expresses at least Fc ⁇ RIII and performs ADCC effector functions.
  • human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils.
  • PBMCs peripheral blood mononuclear cells
  • NK natural killer cells
  • monocytes cytotoxic T cells
  • neutrophils effector cells can be separated from natural sources (e.g., blood). Effector cells are generally lymphocytes associated with the effector phase and are used to produce cytokines (helper T cells), cells that kill infectious pathogens (cytotoxic T cells), or secrete antibodies (differentiated B cells).
  • Figure 1 depicts an exemplary bispecific antibody, in which a full-length antibody capable of specifically recognizing a first antigen (BAFFR) is fused with a second antigen (BAFFR) binding portion and a third antigen (CD3) binding portion, wherein the second antigen binding portion is Fab, and the second antigen binding portion is fused to the N-terminus of a heavy chain of the first antigen binding portion, and the third antigen binding portion is scFv, and the third antigen binding portion replaces the Fab region of the first antigen binding portion fused with the second antigen binding portion, and the first Fc region of the bispecific antibody is knob-Fc, and the second Fc region is hole-Fc.
  • BAFFR full-length antibody capable of specifically recognizing a first antigen
  • BAFFR second antigen binding portion
  • CD3 antigen (CD3) binding portion CD3 binding portion
  • the second antigen binding portion is Fab
  • the second antigen binding portion is fused to the N-terminus of a heavy chain of
  • Figure 2 describes an exemplary bispecific antibody, in which a full-length antibody capable of specifically recognizing a first antigen (BAFFR) is fused with a second antigen (BAFFR) binding portion and a third antigen (CD3) binding portion, wherein the second antigen binding portion is Fab, and the second antigen binding portion is fused to the N-terminus of a heavy chain of the first antigen binding portion, and the third antigen binding portion is Fab, and the VH and VL of the third antigen binding portion are interchanged, and the third antigen binding portion replaces the Fab region of the first antigen binding portion fused with the second antigen binding portion, and the first Fc region of the bispecific antibody is knob-Fc, and the second Fc region is hole-Fc.
  • BAFFR full-length antibody capable of specifically recognizing a first antigen
  • BAFFR second antigen binding portion
  • CD3 antigen (CD3) binding portion CD3 binding portion
  • FIG 3 shows the structure of bispecific antibody C, in which a full-length antibody capable of specifically recognizing a first antigen (BAFFR) is fused with a second antigen (CD3) binding portion, wherein the second antigen binding portion is Fab, the VH and VL of the second antigen binding portion are interchanged, and the second antigen binding portion replaces the Fab region on one side of the first antigen binding portion, the first Fc region of the bispecific antibody is knob-Fc, and the second Fc region is hole-Fc.
  • BAFFR full-length antibody capable of specifically recognizing a first antigen
  • CD3 second antigen
  • FIG4 shows the binding activity of the bispecific antibody to BAFFR protein.
  • FIG5 shows the binding activity of bispecific antibodies to CD3 protein.
  • FIG6 shows the binding activity of bispecific antibodies to Jeko-1 cells.
  • FIG. 7 shows the binding activity of bispecific antibodies to Raji cells.
  • FIG8 shows the binding activity of bispecific antibodies to Su-DHL4 cells.
  • FIG. 9 shows the T cell-specific activation activity of the bispecific antibody against jukart-NFAT-luc reporter gene cells.
  • FIG. 10 shows the killing activity of the bispecific antibody against CHO-K1-BAFFR cells.
  • FIG. 11 shows the killing activity of the bispecific antibody against CHO-K1 cells.
  • FIG. 12 shows the killing activity of the bispecific antibody against Raji-luc cells.
  • the embodiment is to construct bispecific antibodies targeting BAFFR and CD3 targets according to Figures 1-3, and they are named Antibody A, Antibody B, and Antibody C respectively.
  • the Fc of the antibody amino acid sequence is adjusted to other IgG types, and the desired form of amino acid mutations is further designed in each heavy chain, thereby obtaining the amino acid sequence of the target antibody (see Table 1).
  • the constructed antibody amino acid sequence combination is shown in Table 2 and includes the theoretical molecular weight.
  • the above target amino acid sequences were converted into nucleotide sequences, and optimized for a series of parameters that may affect the expression of antibodies in mammalian cells: codon preference, GC content (i.e., the ratio of guanine G and cytosine C in the four bases of DNA), CpG island (i.e., the region with a higher density of CpG dinucleotides in the genome), secondary structure of mRNA, splicing site, pre-mature PolyA site, internal Chi site (a short DNA fragment in the genome, near which the probability of homologous recombination increases) or ribosome binding site, RNA unstable sequence, inverted repeat sequence and restriction enzyme sites that may interfere with cloning; at the same time, related sequences that may improve translation efficiency, such as Kozak sequence and SD sequence, were added.
  • codon preference i.e., the ratio of guanine G and cytosine C in the four bases of DNA
  • CpG island i.e., the
  • the heavy chain gene and light chain gene encoding the above antibodies were designed, and the nucleotide sequences encoding signal peptides optimized according to the amino acid sequence were designed at the 5' end of the heavy chain and light chain, respectively; in addition, stop codons were added to the 3' end of the light chain and heavy chain nucleotide sequences, respectively.
  • the pcDNA3.1-G418 vector is used as a plasmid vector for expressing the multifunctional antibody.
  • the pcDNA3.1-G418 vector contains the promoter CMVPromoter, the eukaryotic screening marker G418 tag and the prokaryotic screening marker Ampicilline.
  • the nucleotide sequence for constructing the antibody expression light chain and heavy chain is obtained by gene synthesis, and the vector and the target fragment are double-digested with HindIII and XhoI, and then enzymatically linked by DNA ligase after recovery, and transformed into Escherichia coli competent cells DH5 ⁇ , and the positive clones are selected and plasmid extraction and enzyme digestion verification are performed to obtain the antibody-containing plasmid.
  • the recombinant plasmids containing the above-mentioned target genes were transformed into Escherichia coli competent cells DH5 ⁇ , and the transformed bacteria were spread on LB plates containing 100 ⁇ g/mL ampicillin for culture.
  • the plasmid clones were selected and cultured in liquid LB medium, and the bacteria were shaken at 260 rpm for 14 hours.
  • the plasmids were extracted using an endotoxin-free plasmid extraction kit, dissolved in sterile water, and the concentration was measured using a nucleic acid protein quantifier.
  • Expi CHO was cultured at 37°C, 8% CO 2 , 100 rpm to a cell density of 6 ⁇ 10 6 cells/mL.
  • the constructed plasmids were transfected into the above cells according to the combination pairing using liposomes, the transfection plasmid concentration was 1 mg/mL, the liposome volume was determined with reference to the Expi CHO TM Expression System kit, and cultured at 32°C, 5% CO 2 , 100 rpm for 7-10 days. Feed was performed once 18-22 hours after transfection and between the 5th day.
  • the above culture product was centrifuged at 4000g, filtered through a 0.22 ⁇ m filter membrane and the culture supernatant was collected.
  • the antibody protein obtained was purified using Protein A and ion columns and the eluate was collected.
  • the specific operation steps of Protein A and ion column purification are as follows: after high-speed centrifugation of the cell culture fluid, the supernatant is taken and affinity chromatography is performed using GE's Protein A chromatography column.
  • the equilibrium buffer used for chromatography is 1 ⁇ PBS (pH7.4). After the cell supernatant is loaded and bound, it is washed with PBS until the ultraviolet returns to the baseline, and then the target protein is eluted with 0.1M glycine (pH3.0) elution buffer, and the pH is adjusted to neutral with Tris for storage.
  • the pH of the product obtained by affinity chromatography is adjusted to 1-2 pH units lower or higher than pI, and appropriately diluted to control the sample conductivity below 5ms/cm.
  • appropriate corresponding pH buffers such as phosphate buffer, acetate buffer and other conditions
  • conventional ion exchange chromatography methods in the field such as anion exchange or cation exchange are used to perform NaCl gradient elution under corresponding pH conditions, and the collection tubes where the target protein is located are selected according to SDS-PAGE and combined for storage.
  • the purified eluate was ultrafiltered and exchanged into a buffer solution, and the protein was detected by SDS-polyacrylamide gel electrophoresis.
  • Human-BAFFR-His (purchased from Acro, Cat: BAR-H52H3) was diluted to 1 ⁇ g/mL using PBS buffer at pH 7.4, and 100 ⁇ L was added to each well of a 96-well ELISA plate and coated overnight at 4°C. After blocking with 1% BSA blocking solution for 1 hour. After washing the plate 3 times with PBST, the constructed antibody was diluted to 100 nM with 0.5% BSA sample diluent, and this was used as the starting concentration. A 3-fold gradient dilution was performed, with a total of 11 gradients, 100 ⁇ L per well, and incubated at 37°C for 1 hour.
  • the plate was washed 3 times with PBST again, and HRP-labeled goat anti-human IgG-Fc (purchased from Jackson, Cat: 109-035-098) was diluted 1:20000 with sample diluent, 100 ⁇ L was added to each well, and incubated at room temperature for 1 hour. Negative control (irrelevant antibody) and positive control were set up.
  • the positive control was BAFFR monoclonal antibody (BAFFR monoclonal antibody sequence consists of SEQ ID NO: 1 and SEQ ID NO: 2, with the constant region of human IgG1 added, see SEQ ID NO: 27 and SEQ ID NO: 28).
  • 100 ⁇ L TMB substrate was added to each well. Incubate at room temperature in the dark for 10 minutes, and 100 ⁇ L 1M HCl solution was added to each well to terminate the color reaction. Read the plate on a multifunctional microplate reader.
  • the ELISA results of the antibodies are shown in FIG4 , and the bispecific antibodies can bind to the BAFFR protein at all concentrations.
  • Human-CD3-His (purchased from Acro, Cat: CDD-H52W4) was diluted to 0.2 ⁇ g/mL using pH 7.4 PBS buffer, and 100 ⁇ L was added to each well of a 96-well ELISA plate and coated overnight at 4°C. After blocking with 1% BSA blocking solution for 1 hour. After washing the plate 3 times with PBST, the constructed expression antibody was diluted to 100 nM with 0.5% BSA sample diluent, and this was used as the starting concentration for 3-fold gradient dilution, a total of 11 gradients, and negative controls (blank wells and IgG1 isotype controls) and positive controls were set up.
  • the positive control was CD3 monoclonal antibody (CD3 antibody sequence was derived from INN blinatumomab, the sequence consisted of SEQ ID NO: 9, SEQ ID NO: 10, and the constant region of human IgG1 was added.
  • the constant region of IgG1 is shown in SEQ ID NO: 27 and SEQ ID NO: 28), 100 ⁇ L per well, incubated at 37°C for 1 hour. Wash the plate three times with PBST, dilute HRP-labeled goat anti-human IgG-Fc with sample diluent at 1:20,000, add 100 ⁇ L to each well, and incubate at room temperature for 1 hour.
  • the ELISA results of the antibodies are shown in FIG5 , and the bispecific antibodies can bind to CD3 at all concentrations.
  • Example 7 Flow cytometry detection of antibody binding activity to BAFFR-positive tumor cells
  • the positive control is BAFFR monoclonal antibody (BAFFR monoclonal antibody sequence consists of SEQ ID NO: 1 and SEQ ID NO: 2, and the constant region of human IgG1 is added.
  • the constant region of human IgG1 is shown in SEQ ID NO: 27 and SEQ ID NO: 28), and add 100 ⁇ L of antibody diluent.
  • the cells were incubated at 4°C for 60 minutes and then washed twice with excess FACS buffer.
  • the cells were resuspended in 100 ⁇ L FACS buffer, and anti-human IgG Fc fluorescent secondary antibody-FITC (Biolegend, Cat: 109306) was added to the sample, incubated for 30 minutes and washed twice with excess FACS buffer.
  • the cells were resuspended in flow cytometry buffer and then detected and analyzed by flow cytometry.
  • BAFFR-positive Jeko-1 cells (from the Chinese Academy of Sciences Cell Collection Center) were plated in 96-well plates at 2 ⁇ 10 4 /well, and then the effector cells Jurkat-NFAT-Luc were added at 5 ⁇ 10 4 /well.
  • the antibody was diluted to 30 ⁇ g/mL using PBS, diluted 5 times, and a total of 9 concentration gradients were added to the wells of the corresponding cells.
  • a negative control group was set up and cultured at 37°C for 6h. Then Bio-Lite was added to the sample wells and incubated at room temperature for 10min before reading with a multifunctional microplate reader.
  • bispecific antibodies A and B specifically activate jurkart-NFAT-Luc reporter cells expressing CD3 in the presence of BARRF-positive target cells Jeko-1. However, there is no activation activity in the absence of antibodies or target cells. This shows that after bispecific antibodies A and B recognize BAFFR-positive cells, they can specifically activate CD3-positive T cells, and then activate downstream signals that induce T cells to kill tumors.
  • Bispecific antibody C has non-specific activation and poses risks.
  • BAFFR-positive CHO-K1-BAFFR cells BAFFR overexpression engineered cell line constructed by BAFFR lentiviral transduction of CHO-K1 cells
  • BAFFR-negative CHO-K1 BAFFR-negative CHO-K1
  • bispecific antibodies and irrelevant antibodies were added, starting at 10 ⁇ g/mL and diluted 10 times, with a total of 6 concentration gradients.
  • CIK CD3+CD56+ cells
  • effector cells were added at 1 ⁇ 10 5 /well, with an effector-target ratio of 5:1. Blank control (diluent), negative control (target cells + CIK cells, no antibody), and irrelevant antibody group were set.
  • negative control 1 was BAFFR monoclonal antibody (BAFFR monoclonal antibody sequence consists of SEQ ID NO: 1 and SEQ ID NO: 2, with the constant region of human IgG1 added, and the constant region of human IgG1 is shown in SEQ ID NO: 27 and SEQ ID NO: 28), and negative control 2 was anti-GPC3 and CD3 antibody (the antibody is derived from patent US11001643B2, and the amino acid sequence consists of SEQ ID NO: 29, SEQ ID NO: 30 and SEQ ID NO: 31). After incubation in a cell culture incubator for 24 h, the cells were rinsed with PBS for 3 h.
  • BAFFR monoclonal antibody sequence consists of SEQ ID NO: 1 and SEQ ID NO: 2, with the constant region of human IgG1 added, and the constant region of human IgG1 is shown in SEQ ID NO: 27 and SEQ ID NO: 28
  • negative control 2 was anti-GPC3 and CD3 antibody (the antibody is derived from patent US110016
  • Cell killing rate (%) (1-(OD value of sample well-OD value of blank well)/(OD value of negative well-OD value of blank well) OD value)) ⁇ 100%.
  • the bispecific antibody can kill CHO-K1-BAFFR overexpressing BAFFR, and the irrelevant antibody has no killing effect. Both the bispecific antibody and the irrelevant antibody have no killing effect on CHO-K1, indicating that the bispecific antibody can mediate the specific killing of BAFFR-positive cells by CIK cells.
  • BAFFR-positive Raji-luc (Luc lentiviral transduction of raji) was plated in 96-well plates at 1 ⁇ 10 4 /well. After culturing for 24 hours, bispecific antibodies and irrelevant antibodies were added, starting at 50 ⁇ g/mL, 10-fold dilution, a total of 6 concentration gradients, and CIK (CD3+CD56+ cells) effector cells were added at 1 ⁇ 10 5 /well, with an effector-target ratio of 10:1, and blank controls (diluent), negative controls (target cells + CIK cells, no antibodies), and irrelevant antibody groups were set.
  • negative control 1 was BAFFR monoclonal antibody (BAFFR monoclonal antibody sequence consists of SEQ ID NO:1, SEQ ID NO:2, and the constant region of human IgG1 is added, and the constant region of IgG1 is shown in SEQ ID NO:27 and SEQ ID NO:28), and negative control 2 was anti-GPC3 and CD3 bispecific antibodies (the antibody sequence is derived from patent US11001643B2, and the sequence consists of SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31).
  • Bio-Lite was added, and after incubation in the incubator for 10 minutes, the OD value was detected at 450nm on a microplate reader.
  • the bispecific antibody can kill BAFFR-positive Raji, while the irrelevant antibody has no killing effect, indicating that the bispecific antibody mediates the CIK cell-specific killing of BAFFR-positive cells.

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Abstract

L'invention concerne un anticorps bispécifique, qui comprend : (a) une première partie de liaison à l'antigène qui se lie spécifiquement à un premier antigène, le premier antigène étant BAFFR ; (b) une deuxième partie de liaison à l'antigène qui se lie spécifiquement à un deuxième antigène, le deuxième antigène étant BAFFR ; et (c) une troisième partie de liaison à l'antigène qui se lie spécifiquement à un troisième antigène, le troisième antigène étant CD3. L'anticorps bispécifique maximise l'effet de destruction tumorale par liaison multivalente d'antigènes associés à une tumeur tout en régulant la toxicité de CD3 : un anticorps BAFFR se lie de manière multivalente à une tumeur positive à BAFFR et un anticorps CD3 à l'autre extrémité se lie à un lymphocyte T ; en tant qu'adaptateur, l'anticorps bispécifique amène le lymphocyte T et une cellule tumorale plus proche à former une synapse immunitaire, de telle sorte que le lymphocyte T tue la tumeur ; l'anticorps CD3 monovalent réduit la toxicité et peut mieux former une synapse immunitaire pour obtenir une réaction antitumorale plus efficace, plus spécifique et plus sûre.
PCT/CN2023/123288 2022-10-08 2023-10-08 Anticorps bispécifique se liant à baffr et cd3 et utilisation associée WO2024074145A1 (fr)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106589129A (zh) * 2016-12-30 2017-04-26 上海近岸生物科技有限公司 一种结合cd19、cd3和cd28的三功能分子及其应用
WO2020135201A1 (fr) * 2018-12-28 2020-07-02 四川科伦博泰生物医药股份有限公司 Anticorps et son utilisation
US20200339686A1 (en) * 2018-01-16 2020-10-29 Lakepharma, Inc. Bispecific antibody that binds cd3 and another target
US20200362054A1 (en) * 2017-11-21 2020-11-19 Brian Granda Trispecific binding molecules against tumor-associated antigents and use thereof
CN113412124A (zh) * 2018-12-19 2021-09-17 希望之城 Baff-r双特异性t细胞衔接子抗体
CN114728065A (zh) * 2019-11-26 2022-07-08 上海岸迈生物科技有限公司 针对cd3和bcma的抗体和自其制备的双特异性结合蛋白

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106589129A (zh) * 2016-12-30 2017-04-26 上海近岸生物科技有限公司 一种结合cd19、cd3和cd28的三功能分子及其应用
US20200362054A1 (en) * 2017-11-21 2020-11-19 Brian Granda Trispecific binding molecules against tumor-associated antigents and use thereof
US20200339686A1 (en) * 2018-01-16 2020-10-29 Lakepharma, Inc. Bispecific antibody that binds cd3 and another target
CN113412124A (zh) * 2018-12-19 2021-09-17 希望之城 Baff-r双特异性t细胞衔接子抗体
WO2020135201A1 (fr) * 2018-12-28 2020-07-02 四川科伦博泰生物医药股份有限公司 Anticorps et son utilisation
CN114728065A (zh) * 2019-11-26 2022-07-08 上海岸迈生物科技有限公司 针对cd3和bcma的抗体和自其制备的双特异性结合蛋白

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
WU, L. J. ET AL.: "Bispecific BCMA-CD3 Antibodies Block Multiple Myeloma Tumor Growth", CANCERS, vol. 14, 20 May 2022 (2022-05-20), XP093095867, DOI: 10.3390/cancers14102518 *

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