WO2023193767A1 - Protéine de liaison à l'antigène et son application - Google Patents

Protéine de liaison à l'antigène et son application Download PDF

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Publication number
WO2023193767A1
WO2023193767A1 PCT/CN2023/086611 CN2023086611W WO2023193767A1 WO 2023193767 A1 WO2023193767 A1 WO 2023193767A1 CN 2023086611 W CN2023086611 W CN 2023086611W WO 2023193767 A1 WO2023193767 A1 WO 2023193767A1
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Prior art keywords
antigen
binding protein
seq
amino acid
acid sequence
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PCT/CN2023/086611
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English (en)
Chinese (zh)
Inventor
邓俗俊
刘培培
顾春银
王宗达
曹晓丹
杨欣秀
Original Assignee
上海济煜医药科技有限公司
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Publication of WO2023193767A1 publication Critical patent/WO2023193767A1/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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes

Definitions

  • This application relates to the field of biomedicine, specifically to an antigen-binding protein and its application.
  • Programmed death 1 (programmed death 1), referred to as PD-1, is widely expressed in immune cells and is an important immunosuppressive molecule.
  • the main ligand of PD-1 is PD-L1, and PD-L1 is mainly expressed on the surface of tumor cells. After the ligand PD-L1 binds to the receptor PD-1, it inhibits T cell activation in the tumor microenvironment, causing the immune system such as T cells to be unable to kill tumor cells normally, thereby achieving immune escape.
  • the present application provides an antigen-binding protein, which may have one or more of the following properties: (1) capable of binding to primate-derived PD- L1 protein; and (2) can stimulate immune cells to secrete cytokines, for example, stimulate lymphocytes to secrete IL-2.
  • the present application provides an antigen-binding protein comprising an antigen-binding fragment capable of binding PD-L1.
  • the antigen-binding protein includes an antibody heavy chain variable region VH
  • the VH includes HCDR1, HCDR2 and HCDR3
  • the HCDR3 includes the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 includes SEQ ID NO: 5
  • the amino acid sequence shown is, and the HCDR1 includes the amino acid sequence shown in SEQ ID NO: 1.
  • the antigen-binding protein includes an antibody light chain variable region VL
  • the VL includes LCDR1, LCDR2 and LCDR3
  • the LCDR3 includes the amino acid sequence shown in SEQ ID NO: 25
  • the LCDR2 includes SEQ ID NO: 24
  • the amino acid sequence shown is, and the LCDR1 includes the amino acid sequence shown in SEQ ID NO: 23.
  • the present application provides a polypeptide, the antigen-binding protein comprising a first targeting moiety capable of binding to PD-L1 and a second targeting moiety capable of binding to PD-1.
  • the first targeting moiety includes the PD-L1 antigen-binding protein described in any one of the present applications.
  • the second targeting portion includes the antibody heavy chain variable region VH
  • the VH of the second targeting portion includes HCDR1, HCDR2 and HCDR3
  • the HCDR3 of the second targeting portion includes SEQ ID NO: 14
  • HCDR2 of the second targeting part includes the amino acid sequence shown in SEQ ID NO: 13 acid sequence
  • the HCDR1 of the second targeting portion includes the amino acid sequence shown in SEQ ID NO: 12.
  • the second targeting portion includes an antibody light chain variable region VL
  • the VL of the second targeting portion includes LCDR1, LCDR2, and LCDR3, and the LCDR3 of the second targeting portion includes SEQ ID NO: 25.
  • the LCDR2 of the second targeting portion includes the amino acid sequence shown in SEQ ID NO: 24, and the LCDR1 of the second targeting portion includes the amino acid sequence shown in SEQ ID NO: 23.
  • the polypeptide is a multispecific antibody with a common light chain, such as a bispecific antibody.
  • the present application provides a nucleic acid encoding the antigen-binding protein of the present application and/or the polypeptide of the present application.
  • the present application provides a vector comprising the nucleic acid of the present application.
  • the present application provides a cell comprising the antigen-binding protein of the present application, the polypeptide of the present application, and/or the nucleic acid of the present application.
  • the present application provides a method for preparing the antigen-binding protein of the present application and/or the polypeptide of the present application, which method includes culturing under conditions that allow the expression of the antigen-binding protein and/or the polypeptide. cells according to the present application.
  • the present application provides a conjugate comprising an antigen-binding protein of the present application and/or a polypeptide of the present application.
  • the present application provides a pharmaceutical composition comprising the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cell of the present application, and/or The conjugates of the present application, and optionally a pharmaceutically acceptable carrier.
  • the present application provides a kit, which includes the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cells of the present application, and the conjugation of the present application. substances, and/or pharmaceutical compositions of the present application.
  • the present application provides a method for influencing cells to produce cytokines, the method comprising administering the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cells of the present application, The conjugate of the present application, the pharmaceutical composition of the present application, and/or the kit of the present application.
  • the present application provides a method for promoting cell bridging, the method comprising administering the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cell of the present application, the conjugate of the present application, the The pharmaceutical composition, and/or the kit of the present application.
  • the present application provides the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cell of the present application, the conjugate of the present application, the pharmaceutical composition of the present application, and /or the kit of this application Use in the preparation of medicaments for preventing, alleviating and/or treating tumors.
  • the present application provides the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cell of the present application, the conjugate of the present application, the pharmaceutical composition of the present application, and / Or the kit of the present application, which is used to prevent, alleviate and / or treat tumors.
  • the present application provides a method for preventing, alleviating and/or treating tumors, comprising administering the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cells of the present application, The conjugate of the present application, the pharmaceutical composition of the present application, and/or the kit of the present application.
  • Figure 1 shows the activation results of the antibody of the present application on the release of IL-2 from lymphocytes.
  • Figure 2 shows the activation results of the antibody of the present application on the release of IL-2 from lymphocytes.
  • Figure 3 shows the activation results of another antibody of the present application on the release of IL-2 from lymphocytes.
  • Figure 4 shows an exemplary bispecific antibody construction method of the present application.
  • Figure 5 shows the activation results of the bispecific antibody of the present application on the release of IL-2 from lymphocytes.
  • Figure 6 shows the results of the cell bridging function of the bispecific antibody of the present application.
  • Figure 7 shows the inhibitory effect of the bispecific antibody of the present application on tumor growth.
  • PD-L1 generally refers to the programmed death ligand 1 protein.
  • PD-L1 is also known as cluster of differentiation 274 (CD274) or B7 homolog 1 (B7-H1), and is the protein encoded by the CD274 gene (in humans).
  • CD274 cluster of differentiation 274
  • B7-H1 B7 homolog 1
  • PD-L1 can bind to its receptors, such as programmed death 1 (PD-1).
  • PD-1 and PD-1 exerts an immunosuppressive effect by inhibiting T cell proliferation and producing the cytokines IL-2 and IFN- ⁇ .
  • PD-L1 encompasses any native PD-L1 or modified PD-L1 from any vertebrate source, including mammals, such as primates (e.g., humans or monkeys) and rodents ( e.g. mouse or rat).
  • mammals such as primates (e.g., humans or monkeys) and rodents (e.g. mouse or rat).
  • the term encompasses "full-length", unprocessed PD-L1 as well as any form of PD-L1 resulting from processing in the cell.
  • PD-L1 can exist as a transmembrane protein or as a soluble protein.
  • the term also encompasses naturally occurring variants of PD-L1, such as splice variants or allelic variants.
  • the basic structure of PD-L1 includes four domains: extracellular Ig-like V-type domain and Ig-like C2-type domain, transmembrane domain and cytoplasmic domain.
  • PD-L1 sequences are known in the art. Information on the human PD-L1 gene, including the genomic DNA sequence, can be found, for example, under NCBI Gene ID No. 29126.
  • the amino acid sequence of an exemplary full-length human PD-L1 protein can be found under UniProt accession number Q9NZQ7.
  • PD-1 generally refers to the programmed death 1 receptor, which may also be referred to as “programmed death 1", “CD279”, “cluster of differentiation 279", “PD1", “PDCD1” or “CD297”.
  • PD-1 proteins usually include an extracellular IgV domain, a transmembrane region and an intracellular tail.
  • PD-1 is commonly expressed on T cells, B cells, natural killer T cells, activated monocytes, and dendritic cells (DC).
  • DC dendritic cells
  • PD-1 can bind to its ligands PD-L1 and PD-L2.
  • PD-1 encompasses any native PD-1 or modified PD-1 from any vertebrate source, including mammals, such as primates (e.g., humans or monkeys) and rodents ( e.g. mouse or rat).
  • the term encompasses "full-length", unprocessed PD-1 as well as any form of PD-1 resulting from processing in the cell.
  • PD-1 can exist as a transmembrane protein or as a soluble protein.
  • PD-1 includes complete PD-1 and its fragments, and also includes functional variants, isoforms, species homologs, derivatives, analogs of PD-1, and those that have at least one property in common with PD-1 Analogues of epitopes. PD-1 sequences are known in the art.
  • an exemplary full-length human PD-1 protein sequence can be found under NCBI accession number NP_005009.2, and an exemplary full-length cynomolgus monkey PD-1 protein sequence can be found under NCBI accession number NP_001271065 or UniProt accession number BOLAJ3 turn up.
  • antigen-binding protein generally refers to a protein comprising an antigen-binding portion, and optionally a scaffold or backbone portion that allows the antigen-binding portion to adopt a conformation that promotes binding of the antigen-binding protein to the antigen.
  • Antigen binding proteins may typically comprise an antibody light chain variable region (VL), an antibody heavy chain variable region (VH), or both, and functional fragments thereof.
  • VL antibody light chain variable region
  • VH antibody heavy chain variable region
  • the variable regions of the heavy and light chains contain binding domains that interact with the antigen.
  • antigen-binding proteins include, but are not limited to, antibodies, antigen-binding fragments, immunoconjugates, multispecific antibodies (e.g., bispecific antibodies), antibody fragments, antibody derivatives, antibody analogs, or fusion proteins, etc., as long as they display The required antigen-binding activity can be obtained.
  • the term "antibody” generally refers to an immunoglobulin reactive to a specified protein or peptide or fragment thereof.
  • the antibodies may be from any class, including, but not limited to, IgG, IgA, IgM, IgD, and IgE, and from any subclass (eg, IgGl, IgG2, IgG3, and IgG4).
  • the antibody may have a heavy chain constant region selected from, for example, IgGl, IgG2, IgG3, or IgG4.
  • the antibody may also have a light chain selected from, for example, kappa ( ⁇ ) or lambda ( ⁇ ).
  • Antibodies of the present application can be derived from any species.
  • antigen-binding fragment generally refers to a portion of an antibody molecule that contains the amino acid residues that interact with the antigen and confer specificity and affinity to the antibody for the antigen.
  • antigen-binding fragments may include, but are not limited to, Fab, Fab', F(ab) 2 , Fv fragment, F(ab') 2 , scFv, di-scFv and/or dAb.
  • Fab generally refers to a fragment containing the variable domain of the heavy chain and the variable domain of the light chain, and also containing the constant domain of the light chain and the first constant domain (CH1) of the heavy chain.
  • Fab' generally refers to a fragment that is different from Fab by adding a small number of residues (including one or more cysteines from the antibody hinge region) to the carboxyl terminus of the heavy chain CH1 domain
  • F(ab ') 2 usually refers to a dimer of Fab', an antibody fragment containing two Fab fragments connected by a disulfide bridge on the hinge region.
  • Fv generally refers to the smallest antibody fragment containing intact antigen recognition and binding sites.
  • the fragment may consist of a heavy chain variable domain and a light chain variable domain as a dimer in tight non-covalent association;
  • dsFv generally refers to a disulfide-stabilized Fv fragment, The bond between its single light chain variable domain and its single heavy chain variable domain is a disulfide bond.
  • dAb fragment generally refers to an antibody fragment consisting of a VH domain.
  • scFv generally refers to a monovalent molecule formed by covalently connecting a heavy chain variable domain and a light chain variable domain of an antibody through a flexible peptide linker; such scFv molecules may have general Structure: NH 2 -VL-linker-VH-COOH or NH 2 -VH-linker-VL-COOH.
  • variable region or “variable domain” generally refers to the domain of an antibody heavy or light chain that is involved in binding of the antibody to an antigen.
  • variable generally means that certain portions of the sequence of the variable domain of an antibody vary strongly, resulting in the binding and specificity of various specific antibodies for their specific antigens. Variability is not evenly distributed throughout the variable regions of an antibody. It is concentrated in three segments in the light chain variable region and heavy chain variable region, known as the complementarity determining region (CDR) or hypervariable region (HVR), which are LCDR1, LCDR2, LCDR3, HCDR1, HCDR2 and HCDR3. The more highly conserved portions of the variable domains are called framework regions (FR).
  • CDR complementarity determining region
  • HVR hypervariable region
  • variable domains of the native heavy and light chains each contain four FR regions (H-FR1, H-FR2, H-FR3, H-FR4, L-FR1, L-FR2, L-FR3, L-FR4) , most adopt ⁇ -sheet configuration and are connected through three CDR structural loop regions.
  • the CDRs in each chain are held closely together by the FR region and, together with the CDRs from the other chain, form the antigen-binding site of the antibody.
  • variable regions of an antibody can be encoded or the CDRs of an antibody can be delineated by a variety of methods, such as the Kabat numbering scheme and definition rules based on sequence variability (see, Kabat et al., Protein Sequences in Immunology, 5 ed., National Institutes of Health, Bethesda, MD (1991)), based on the location of structural loop regions.
  • the term "monoclonal antibody” generally refers to an antibody obtained from a population of antibodies that are essentially homogeneous, i.e., the individual antibodies making up the population are identical except for possible naturally occurring mutations that may be present in minimal amounts and/or In addition to post-translational modifications (such as isomerization, amidation). Monoclonal antibodies are highly specific and target a single antigenic site.
  • chimeric antibody generally refers to an antibody in which the variable regions are derived from one species and the constant regions are derived from another species.
  • the variable regions are derived from an antibody from a laboratory animal, such as a rodent (a "parent antibody”), and the constant regions are derived from a human antibody, such that the resulting chimeric antibody is more effective in a human individual than the parent (eg, mouse-derived) antibody. Less likely to trigger an adverse immune response.
  • humanized antibody generally refers to an antibody in which some or all of the amino acids outside the CDR region of a non-human antibody (eg, a mouse antibody) are replaced with corresponding amino acids derived from human immunoglobulins. Additions, deletions, insertions, substitutions or modifications of amino acids in the CDR regions may also be allowed as long as they retain the ability of the antibody to bind a specific antigen.
  • the humanized antibody may optionally comprise at least a portion of a human immunoglobulin constant region. "Humanized antibodies” retain antigen specificity similar to the original antibody.
  • “Humanized” forms of non-human (eg, murine) antibodies may minimally comprise chimeric antibodies derived from sequences derived from non-human immunoglobulins.
  • CDR region residues in a human immunoglobulin can be used with a non-human species (donor antibody) having the desired properties, affinity, and/or ability (such as mouse, rat , rabbit or non-human primate) CDR region residue substitution.
  • donor antibody non-human species having the desired properties, affinity, and/or ability (such as mouse, rat , rabbit or non-human primate) CDR region residue substitution.
  • FR region residues of a human immunoglobulin can be replaced with corresponding non-human residues.
  • humanized antibodies may contain amino acid modifications that are not present in the recipient antibody or in the donor antibody.
  • the term "fully human antibody” generally refers to an antibody in which all parts (including the variable and constant regions of the antibody) are encoded by genes of human origin.
  • Methods for obtaining fully human antibodies in this field include phage display technology, transgenic mouse technology, ribosome display technology, and RNA-polypeptide technology.
  • binding generally refer to a measurable and reproducible interaction, such as binding between an antigen and an antibody, which can be determined in the presence of a molecule
  • a target in a heterogeneous population (including biological molecules).
  • an antibody binds to an epitope through its antigen-binding domain, and this binding requires some complementarity between the antigen-binding domain and the epitope.
  • an antibody that specifically binds a target is An antibody that binds this target with greater affinity, avidity, more readily, and/or for a greater duration than it binds other targets.
  • An antibody is said to "specifically bind" an antigen when it binds to an epitope more readily through its antigen-binding domain than it would to a random, unrelated epitope.
  • KD KD
  • KD KD
  • KD is the dissociation rate constant (kdis, also known as “off-rate”. )(koff)” or “kd”) to the binding rate constant (kon, also known as “binding rate (kon)” or “ka”).
  • the binding affinity of an antigen-binding protein (eg, an antibody) for an antigen can be expressed using the association rate constant (kon), the dissociation rate constant (kdis), and the equilibrium dissociation constant (KD).
  • association and dissociation rate constants are well known in the art, including but not limited to biofilm interference technique (BLI), radioimmunoassay (RIA), equilibrium dialysis, surface plasmon resonance (SPR), fluorescence resonance energy transfer (FRET) , co-immunoprecipitation (Co-IP) and protein chip technology.
  • BBI biofilm interference technique
  • RIA radioimmunoassay
  • SPR surface plasmon resonance
  • FRET fluorescence resonance energy transfer
  • Co-IP co-immunoprecipitation
  • the measured affinity for a particular protein-protein interaction can differ if measured under different conditions (eg, salt concentration, pH).
  • the term "primate” generally refers to aye-aye and ape species, and includes monkey species, such as those from the genus Macaca (e.g., Macaca fascicularis and/or rhesus monkeys (Macaca mulatta)) monkeys and baboons (Papio ursinus), as well as marmosets (species from the genus Callithrix), squirrel monkeys (species from the genus Saimiri) and tamarins (from the genus Tamarinus) Saguinus), as well as ape species such as chimpanzees (Pan troglodytes), and also includes Homo sapiens.
  • monkey species such as those from the genus Macaca (e.g., Macaca fascicularis and/or rhesus monkeys (Macaca mulatta)) monkeys and baboons (Papio ursinus), as well as marmosets (species from
  • polypeptide or "protein” are used interchangeably and generally refer to a polymer of amino acid residues.
  • the term also applies to amino acid polymers in which one or more amino acid residues are analogs or mimetics of the corresponding naturally occurring amino acids, as well as naturally occurring amino acid polymers.
  • the term may also include modified amino acid polymers, for example, by the addition of sugar residues to form glycoproteins or by phosphorylation.
  • Polypeptides and proteins may be produced from naturally occurring and non-recombinant cells or from genetically engineered or recombinant cells, and may comprise molecules having the amino acid sequence of the native protein, or having the deletion, addition, or deletion of one or more amino acids of the native sequence. and/or substituted molecules.
  • polypeptide and protein particularly include sequences in which one or more amino acids of the antigen-binding proteins described herein are deleted, added and/or substituted.
  • a polypeptide of the present application may comprise an antigen-binding protein.
  • a polypeptide of the present application may comprise a multispecific antigen-binding protein, such as a bispecific antibody.
  • isolated generally refers to biological material (eg, viruses, nucleic acids, or proteins) that is substantially free of components that normally accompany or interact with it in its naturally occurring environment.
  • the isolated biological material optionally contains additional materials that the biological material is not found to have in its natural environment (eg, nucleic acids or proteins).
  • isolated when referring to a protein, “isolated” generally refers to the removal of the molecule from the entire organism in which it is found naturally occurring. Isolation and separation, or the substantial absence of other biological macromolecules of the same type. When it comes to a nucleic acid molecule, it is completely or partially separated from the sequence to which it is naturally associated, or the nucleic acid has heterologous sequences to which it is associated, or the nucleic acid is separated from the chromosome.
  • immunoconjugate generally refers to a substance formed by connecting an antigen-binding protein to other active agents.
  • the other active agents can be small molecule active agents, such as chemotherapeutic agents, toxins, immunotherapeutic agents, and imaging probes. or spectroscopic probes.
  • nucleic acid generally refers to an isolated form of nucleotides, deoxyribonucleotides or ribonucleotides or analogs thereof of any length, isolated from their natural environment or artificially synthesized.
  • the term "vector” generally refers to a nucleic acid molecule capable of self-replication in a suitable host, which transfers the inserted nucleic acid molecule into and/or between host cells.
  • the vectors may include vectors primarily used for insertion of DNA or RNA into cells, vectors primarily used for replication of DNA or RNA, and vectors primarily used for expression of transcription and/or translation of DNA or RNA.
  • the vectors also include vectors having a variety of the above-mentioned functions.
  • the vector may be a polynucleotide capable of being transcribed and translated into a polypeptide when introduced into a suitable host cell.
  • the vector can produce the desired expression product by culturing a suitable host cell containing the vector.
  • the term "cell” generally refers to an individual cell, cell line or cell culture that can contain or has contained a plasmid or vector including a nucleic acid molecule described herein, or is capable of expressing an antigen-binding protein described herein. things.
  • the cells may include progeny of a single host cell. Due to natural, accidental or intentional mutations, the progeny cells may not necessarily be identical in morphology or genome to the original parent cells, but they may be able to express the antibodies or antigen-binding fragments thereof described in this application.
  • the cells can be obtained by transfecting cells in vitro using the vectors described in this application.
  • the cells may be prokaryotic cells (e.g.
  • Escherichia coli or eukaryotic cells
  • eukaryotic cells e.g. yeast cells, e.g. COS cells, Chinese hamster ovary (CHO) cells, HeLa cells, HEK293 cells, COS-1 cells, NSO cells or myeloma cells.
  • the cells may be mammalian cells.
  • the mammalian cells may be CHO-K1 cells.
  • the term "pharmaceutical composition” generally refers to a formulation that allows the biological activity of the active ingredient to be present in a form that is effective and does not contain additional ingredients that would have unacceptable toxicity to the subject to whom the composition is to be administered.
  • treatment generally refers to a clinical intervention intended to alter the natural course of the disease in the individual being treated, and may be to achieve prevention or treatment during the clinical course of the disease.
  • Desirable therapeutic effects include, but are not limited to, preventing the onset or recurrence of disease, alleviating symptoms, attenuating any direct or indirect pathological consequences of disease, preventing metastasis, reducing the rate of disease progression, ameliorating or relieving disease status, and alleviating or improving prognosis.
  • antibodies can be used to delay disease development or slow disease progression.
  • administering generally refers to administering to a subject (eg, a patient) a dose of a compound (eg, an anti-cancer therapeutic agent) or a pharmaceutical composition (eg, a pharmaceutical composition comprising an anti-cancer therapeutic agent) Methods. Apply Administration may be by any suitable means, including parenteral, intrapulmonary and intranasal, and, if required for local treatment, intralesional administration. Parenteral infusion includes, for example, intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
  • tumor generally refers to all neoplastic cell growth and proliferation (whether malignant or benign) and all precancerous and cancerous cells and tissues.
  • the tumor may be a tumor with high expression of PD-1 or PD-L1 in cells and tissues.
  • Tumors may include solid tumors and/or non-solid tumors.
  • homologue generally refers to an amino acid sequence or a nucleotide sequence that has certain homology to a wild-type amino acid sequence and a wild-type nucleotide sequence.
  • the term “homology” may be equated with sequence "identity.”
  • homologous sequences may include amino acid sequences that may be at least 80%, 85%, 90%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% identical to the subject sequence. .
  • homologs will contain active sites, etc. that are identical to the subject amino acid sequence.
  • Homology can be considered in terms of similarity (ie, amino acid residues with similar chemical properties/functions), or homology can be expressed in terms of sequence identity.
  • reference to a sequence having a percent identity with any one of the SEQ ID NOs of an amino acid sequence or a nucleotide sequence means that the sequence has said percent identity over the entire length of the SEQ ID NO mentioned. the sequence of.
  • the term "between” usually means that the C-terminus of a certain amino acid fragment is directly or indirectly connected to the N-terminus of the first amino acid fragment, and its N-terminus is directly or indirectly connected to the C-terminus of the second amino acid fragment.
  • indirect connection In the light chain, for example, the N-terminus of the L-FR2 is directly or indirectly connected to the C-terminus of the LCDR1, and the C-terminus of the L-FR2 is directly or indirectly connected to the N-terminus of the LCDR2.
  • the N terminus of the L-FR3 is directly or indirectly connected to the C terminus of the LCDR2, and the C terminus of the L-FR3 is directly or indirectly connected to the N terminus of the LCDR3.
  • the N-terminus of the H-FR2 is directly or indirectly connected to the C-terminus of the HCDR1
  • the C-terminus of the H-FR2 is directly or indirectly connected to the N-terminus of the HCDR2.
  • the N terminus of the H-FR3 is directly or indirectly connected to the C terminus of the HCDR2
  • the C terminus of the H-FR3 is directly or indirectly connected to the N terminus of the HCDR3.
  • the "first amino acid fragment" and the "second amino acid fragment” can be any amino acid fragments that are the same or different.
  • the term “comprises” generally means including, encompassing, containing or encompassing. In some cases, it also means “for” or “composed of”.
  • the term "about” generally refers to a variation within the range of 0.5% to 10% above or below the specified value, such as 0.5%, 1%, 1.5%, 2%, 2.5%, above or below the specified value. 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%.
  • an antigen-binding protein which may have one or more of the following properties: (1) Capable of binding to primate-derived PD-L1 protein with a KD value of 3.56E-09M or lower; and (2) capable of stimulating immune cells to secrete cytokines, for example, stimulating lymphocytes to secrete IL-2.
  • the binding affinity of the primate PD-L1 antigen binding protein to PD-L1 can be determined by any method known in the art. In some cases, binding affinity can be determined by surface plasmon resonance (SPR), enzyme-linked immunoassay (ELISA), binding antigen precipitation, equilibrium dialysis, or biofilm interference (BLI).
  • the binding affinity and K value of the PD-L1 antigen-binding protein to PD-L1 can be determined by biofilm interference (BLI).
  • BLI biofilm interference
  • the ForteBio Octet Molecular Interaction Analyzer can be used to analyze the binding kinetics between antigens and antibodies.
  • the antigen-binding protein of the present application may comprise a multispecific antigen-binding protein, such as a bispecific antibody.
  • the antigen-binding protein of the present application can bind to primate-derived PD-L1 with a KD value of about 3.56E-09M or lower.
  • the value of K D can be about 1E-08M or less, about 9E-09M or less, about 8E-09M or less, about 7E-09M or less, about 6E-09M or less, About 5E-09M or less, about 4E-09M or less, about 3E-09M or less, about 2E-09M or less, about 1E-09M or less, about 5E-10M or less, about 1E - A value of 10 M or less binding to human-derived PD-L1, for example, as detected using the FortieBio Octet Molecular Interaction Analyzer.
  • the antigen-binding protein described in the present application can block the binding of PD-1 to PD-L1.
  • the antigen-binding protein described in the present application may comprise the antibody heavy chain variable region VH, and the antigen-binding protein in the present application may comprise HCDR1, HCDR2 and HCDR3 of the VH shown in any one of SEQ ID NOs: 7 to 11.
  • the CDRs of this application can be divided according to any dividing rules in this field.
  • the antigen-binding protein described in the present application may comprise the antibody heavy chain variable region VH, and the antigen-binding protein in the present application may comprise HFR1, HFR2, HFR3 and HFR4 of the VH shown in any one of SEQ ID NOs: 7 to 11.
  • the antigen-binding protein described in the present application can comprise the antibody heavy chain variable region VH, and the antigen-binding protein in the present application can comprise HCDR1, HCDR2 and HCDR3 of the VH shown in any one of SEQ ID NOs: 7 to 8.
  • the antigen-binding protein of the present application may comprise HCDR1, HCDR2 and HCDR3 of EVHLQQSGAALVKPGASVKMSCKASGYTFTDFWVNWVKQSHGNSLEWIGEIWPNSGATNFNENFKGKATLTVDRSTSTAYLDLTRLTSEDSAIYYCTRELRRPPFTYWGQGASVTVSS (SEQ ID NO: 7).
  • the antigen-binding protein of the present application may comprise the HCDR1, HCDR2 and HCDR3 of QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFWVNWVRQAPGQGLEWIGEIWPNSGATNFNENFKGRATLTVDRSISTAYMELSRLRSDDTAVYYCTRELRRPPFTYWGQGTLVTVSS (SEQ ID NO: 8).
  • the antigen-binding protein described in the present application may comprise an antibody heavy chain variable region VH.
  • the antigen-binding protein of the present application may HCDR1, HCDR2 and HCDR3 of the VH shown in any one of SEQ ID NOs: 9 to 10 may be included.
  • the antigen-binding protein of the present application may comprise HCDR1, HCDR2 and HCDR3 of EVQLVESGSALVKPGASVKMSCKASGYTFTDFWVNWVKQSHGKSLEWIGEIWPNSGTTNFNEKFRGKATLTVDKSTSTAYMELSRLTSEDSAIYYCTRELRRPPFTYWGQGTLVTVSS (SEQ ID NO: 9).
  • the antigen-binding protein of the present application may comprise the HCDR1, HCDR2 and HCDR3 of QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFWVNWVRQAPGQGLEWMGEIWPNSGTTNFAQKFQGRVTMTVDKSISTAYMELSRLRSDDTAVYYCTRELRRPPFTYWGQGTLVTVSS (SEQ ID NO: 10).
  • the antigen-binding protein described in the present application may comprise the antibody heavy chain variable region VH, and the antigen-binding protein described in the present application may comprise HCDR1, HCDR2 and HCDR3 of the VH shown in SEQ ID NO: 11.
  • the antigen-binding protein of the present application may comprise the HCDR1, HCDR2 and HCDR3 of QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFWVNWVRQAPGQGLEWMGEIWPNYGTTNFAQKFQGRVTMTVDKSISTAYMELSRLRSDDTAVYYCTRELRRPPFTYWGQGTLVTVSS (SEQ ID NO: 11).
  • the antigen-binding protein described in the present application may comprise the antibody heavy chain variable region VH, the VH may comprise HCDR1, HCDR2 and HCDR3, and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6.
  • the CDR of this application can be divided according to Chothia division rules.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 5.
  • the HCDR2 may include WPNX 1 GX 2 , where X 1 may be S or Y and X 2 may be A or T.
  • the HCDR2 may comprise at least amino acid substitutions at positions selected from the group consisting of: X 1 and X 2 .
  • the HCDR2 may comprise the amino acid sequence shown in any one of SEQ ID NOs: 2 to 4.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR2 may comprise an amino acid sequence selected from the group consisting of: SEQ ID NO: 2 to 4.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 1.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 5
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 1 Amino acid sequence.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise selected From the amino acid sequence shown in the following group: SEQ ID NO: 2 to 4
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 1.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise SEQ The amino acid sequence shown in ID NO: 2
  • the HCDR1 may include the amino acid sequence shown in SEQ ID NO: 1.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise SEQ The amino acid sequence shown in ID NO: 3
  • the HCDR1 may include the amino acid sequence shown in SEQ ID NO: 1.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise SEQ The amino acid sequence shown in ID NO: 4
  • the HCDR1 may include the amino acid sequence shown in SEQ ID NO: 1.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH, and the VH may comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 7 to 11.
  • the antigen binding protein may comprise an antibody heavy chain constant region CH, and the CH is derived from an IgG constant region.
  • the antigen binding protein may comprise an antibody heavy chain constant region CH, and the CH is derived from an IgGl constant region.
  • the antigen-binding protein may comprise an antibody heavy chain constant region CH, and the CH may comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 16 to 18.
  • the antigen-binding protein may comprise an antibody heavy chain, and the heavy chain may comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 19 to 21.
  • the antigen-binding protein described in the present application may comprise the antibody light chain variable region VL, and the antigen-binding protein in the present application may comprise LCDR1, LCDR2 and LCDR3 of the VL shown in SEQ ID NO: 26.
  • the CDRs of this application can be divided according to any dividing rules in this field.
  • the antigen-binding protein described in the present application can comprise the light chain variable region VL, and the antigen-binding protein in the present application can comprise LFR1, LFR2, LFR3 and LFR4 of the VL shown in SEQ ID NO: 26.
  • the antigen-binding protein described in the present application can comprise the light chain variable region VL
  • the antigen-binding protein in the present application can comprise LCDR1, LCDR2 and LCDR3 of DIQLTQSPSFLSASVGDRVTITCKASQDVGTAVAWYQQKPGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPWTFGGGTKVEIK (SEQ ID NO: 26).
  • the antigen-binding protein may comprise an antibody light chain variable region VL
  • the VL may comprise LCDR1, LCDR2 and LCDR3
  • the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 25.
  • the CDR of this application can be divided according to Chothia division rules.
  • the antigen-binding protein may comprise an antibody light chain variable region VL
  • the VL may comprise LCDR1, LCDR2 and LCDR3
  • the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 24.
  • the antigen-binding protein may comprise an antibody light chain variable region VL
  • the VL may comprise LCDR1, LCDR2 and LCDR3
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 23.
  • the antigen-binding protein may comprise an antibody light chain variable region VL
  • the VL may comprise LCDR1, LCDR2 and LCDR3
  • the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 25
  • the LCDR2 may comprise SEQ The amino acid sequence shown in ID NO: 24
  • the LCDR1 may include the amino acid sequence shown in SEQ ID NO: 23.
  • the antigen-binding protein may comprise an antibody light chain variable region VL, and the VL may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the antigen-binding protein may comprise an antibody light chain constant region CL, and the CL may comprise the amino acid sequence shown in SEQ ID NO: 27.
  • the antigen-binding protein may comprise an antibody light chain, and the light chain may comprise the amino acid sequence shown in SEQ ID NO: 28.
  • the antigen-binding protein described in the present application may comprise the antibody heavy chain variable region VH.
  • the antigen-binding protein in the present application may comprise HCDR1, HCDR2 and HCDR3 of the VH shown in any one of SEQ ID NO: 7 to 11, and the present invention
  • the antigen-binding protein described in the application can include the antibody light chain variable region VL.
  • the antigen-binding protein of the application can include LCDR1, LCDR2 and LCDR3 of the VL shown in SEQ ID NO: 26.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6, the HCDR2 It may comprise an amino acid sequence selected from the following group: SEQ ID NO: 2 to 4, and the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 1
  • the antigen-binding protein may comprise an antibody light chain variable region VL
  • the VL may include LCDR1, LCDR2 and LCDR3
  • the LCDR3 may include the amino acid sequence shown in SEQ ID NO: 25
  • the LCDR2 may include the amino acid sequence shown in SEQ ID NO: 24
  • the LCDR1 may Contains the amino acid sequence shown in SEQ ID NO: 23.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise SEQ The amino acid sequence shown in ID NO: 2
  • the HCDR1 can include the amino acid sequence shown in SEQ ID NO: 1
  • the antigen-binding protein can include the antibody light chain variable region VL
  • the VL can include LCDR1, LCDR2 and LCDR3
  • the LCDR3 may include the amino acid sequence shown in SEQ ID NO:25
  • the LCDR2 may include the amino acid sequence shown in SEQ ID NO:24
  • the LCDR1 may include the amino acid sequence shown in SEQ ID NO:23 sequence.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise SEQ The amino acid sequence shown in ID NO: 3
  • the HCDR1 can include the amino acid sequence shown in SEQ ID NO: 1
  • the antigen-binding protein can include the antibody light chain variable region VL
  • the VL can include LCDR1, LCDR2 and LCDR3
  • the LCDR3 may include the amino acid sequence shown in SEQ ID NO:25
  • the LCDR2 may include the amino acid sequence shown in SEQ ID NO:24
  • the LCDR1 may include the amino acid sequence shown in SEQ ID NO:23 sequence.
  • the antigen-binding protein may comprise an antibody heavy chain variable region VH
  • the VH may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise SEQ The amino acid sequence shown in ID NO: 4
  • the HCDR1 can include the amino acid sequence shown in SEQ ID NO: 1
  • the antigen-binding protein can include the antibody light chain variable region VL
  • the VL can include LCDR1, LCDR2 and LCDR3
  • the LCDR3 may include the amino acid sequence shown in SEQ ID NO:25
  • the LCDR2 may include the amino acid sequence shown in SEQ ID NO:24
  • the LCDR1 may include the amino acid sequence shown in SEQ ID NO:23 sequence.
  • the antigen-binding protein of the present application may comprise HCDR1, HCDR2 and HCDR3, the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6, the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 2, and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6.
  • the HCDR1 may include the amino acid sequence shown in SEQ ID NO: 1; the antigen-binding protein may include LCDR1, LCDR2 and LCDR3, and the LCDR3 may include the amino acid sequence shown in SEQ ID NO: 25 column, the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 24, and the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 23.
  • the antigen-binding protein of the present application may include HCDR1, HCDR2 and HCDR3.
  • the HCDR3 may include the amino acid sequence shown in SEQ ID NO: 6, the HCDR2 may include the amino acid sequence shown in SEQ ID NO: 3, and the HCDR3 may include the amino acid sequence shown in SEQ ID NO: 3.
  • the HCDR1 may include the amino acid sequence shown in SEQ ID NO: 1; the antigen-binding protein may include LCDR1, LCDR2 and LCDR3, the LCDR3 may include the amino acid sequence shown in SEQ ID NO: 25, and the LCDR2 may include SEQ The amino acid sequence shown in ID NO: 24, and the LCDR1 may include the amino acid sequence shown in SEQ ID NO: 23.
  • the antigen-binding protein of the present application may include HCDR1, HCDR2 and HCDR3.
  • the HCDR3 may include the amino acid sequence shown in SEQ ID NO: 6, the HCDR2 may include the amino acid sequence shown in SEQ ID NO: 4, and the HCDR3 may include the amino acid sequence shown in SEQ ID NO: 4.
  • the HCDR1 may include the amino acid sequence shown in SEQ ID NO: 1; the antigen-binding protein may include LCDR1, LCDR2 and LCDR3, the LCDR3 may include the amino acid sequence shown in SEQ ID NO: 25, and the LCDR2 may include SEQ The amino acid sequence shown in ID NO: 24, and the LCDR1 may include the amino acid sequence shown in SEQ ID NO: 23.
  • the antigen-binding protein described in the present application may include the antibody heavy chain variable region VH.
  • the antigen-binding protein described in the present application may include the VH shown in any one of SEQ ID NO: 7 to 11, and the antigen-binding protein described in the present application may include Antibody light chain variable region VL, the antigen-binding protein of the present application can include the VL shown in SEQ ID NO: 26.
  • the antigen-binding protein may bind PD-L1 or a functionally active fragment thereof.
  • the PD-L1 may comprise PD-L1 derived from humans and/or monkeys.
  • the antigen-binding protein may comprise an antibody or antigen-binding fragment thereof.
  • the antibody may be selected from the group consisting of murine antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.
  • the antigen-binding fragment may be selected from the group consisting of Fab, Fab', F(ab) 2 , Fv fragment, F(ab') 2 , scFv, di-scFv, VHH and dAb.
  • the present application provides a polypeptide, which may comprise a first targeting moiety, and the first targeting moiety may comprise an antigen-binding protein of the present application.
  • polypeptide may further comprise a second targeting moiety capable of binding PD-1 or a functionally active fragment thereof.
  • the PD-1 may comprise PD-1 derived from humans and/or monkeys.
  • the second targeting moiety may comprise an antibody or antigen-binding fragment thereof.
  • the antibody may be selected from the group consisting of murine antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.
  • the antigen-binding fragment may be selected from the group consisting of Fab, Fab', F(ab) 2 , Fv fragment, F(ab') 2 , scFv, di-scFv, VHH and dAb.
  • the second targeting portion described in the present application can comprise the antibody heavy chain variable region VH, and the VH of the second targeting portion can comprise HCDR1, HCDR2 and HCDR3 of the VH shown in SEQ ID NO: 15.
  • the CDRs of this application can be divided according to any dividing rules in this field.
  • the second targeting part described in the present application includes the antibody heavy chain variable region VH, and the VH of the second targeting part may include HFR1, HFR2, HFR3 and HFR4 of the VH shown in SEQ ID NO: 15.
  • the antigen-binding protein of the present application may comprise HCDR1, HCDR2 and HCDR3 of EVQLVESGGGLVKPGGSLRLSCAASGFTFSNYDMSWVRQAPGKGLEWVSTISGGGSYTYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCVSPYYGMEYWGQGTLVTVSS (SEQ ID NO: 15).
  • the second targeting portion may comprise an antibody heavy chain variable region VH
  • the VH of the second targeting portion may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 of the second targeting portion may comprise SEQ ID The amino acid sequence shown in NO:14.
  • the CDR of this application can be divided according to Chothia division rules.
  • the second targeting portion may comprise an antibody heavy chain variable region VH
  • the VH of the second targeting portion may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR2 of the second targeting portion may comprise SEQ ID The amino acid sequence shown in NO:13.
  • the second targeting portion may comprise an antibody heavy chain variable region VH
  • the VH of the second targeting portion may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR1 of the second targeting portion may comprise SEQ ID The amino acid sequence shown in NO:12.
  • the second targeting portion may comprise an antibody heavy chain variable region VH, and the VH of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 15.
  • the second targeting moiety may comprise an antibody heavy chain constant region CH, and the CH of the second targeting moiety is derived from an IgG constant region.
  • the second targeting moiety may comprise an antibody heavy chain constant region CH, and the CH of the second targeting moiety is derived from an IgG1 constant region.
  • the second targeting portion may comprise an antibody heavy chain constant region CH, and the CH of the second targeting portion may comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 16 to 18.
  • the second targeting moiety may comprise an antibody heavy chain, and the heavy chain of the second targeting moiety may comprise the amino acid sequence shown in SEQ ID NO: 22.
  • the second targeting part described in the present application may comprise the antibody light chain variable region VL, and the second targeting part in the present application may comprise LCDR1, LCDR2 and LCDR3 of the VL shown in SEQ ID NO: 26.
  • the CDRs of this application can be divided according to any dividing rules in this field.
  • the second targeting part described in the present application may comprise the light chain variable region VL, and the second targeting part in the present application may comprise LFR1, LFR2, LFR3 and LFR4 of the VL shown in SEQ ID NO: 26.
  • the second targeting part described in the present application may comprise the light chain variable region VL
  • the second targeting part in the present application may comprise LCDR1, LCDR2 and LCDR3 of DIQLTQSPSFLSASVGDRVTITCKASQDVGTAVAWYQQKPGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPWTFGGGTKVEIK (SEQ ID NO: 26).
  • the second targeting moiety may comprise an antibody light chain variable region VL
  • the VL of the second targeting moiety may comprise LCDR1, LCDR2 and LCDR3
  • the LCDR3 of the second targeting moiety may comprise SEQ ID The amino acid sequence shown in NO:25.
  • the CDR of this application can be divided according to Chothia division rules.
  • the second targeting portion may comprise an antibody light chain variable region VL
  • the VL of the second targeting portion may comprise LCDR1, LCDR2 and LCDR3
  • the LCDR2 of the second targeting portion may comprise SEQ ID The amino acid sequence shown in NO:24.
  • the second targeting portion may comprise an antibody light chain variable region VL
  • the VL of the second targeting portion may comprise LCDR1, LCDR2 and LCDR3
  • the LCDR1 of the second targeting portion may comprise SEQ ID The amino acid sequence shown in NO:23.
  • the second targeting portion may comprise an antibody light chain variable region VL, and the VL of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the second targeting portion may comprise an antibody light chain constant region CL, and the CL of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 27.
  • the second targeting moiety may comprise an antibody light chain, and the light chain of the second targeting moiety may comprise the amino acid sequence shown in SEQ ID NO: 28.
  • the second targeting portion may comprise an antibody heavy chain variable region VH
  • the VH of the second targeting portion may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 of the second targeting portion may comprise SEQ ID NO. :
  • the amino acid sequence shown in 14, the HCDR2 of the second targeting part may comprise the amino acid sequence shown in SEQ ID NO: 13, and the HCDR1 of the second targeting part may comprise the amino acid sequence shown in SEQ ID NO: 12 Amino acid sequence;
  • the second targeting portion may include the antibody light chain variable region VL, and the VL of the second targeting portion may include LCDR1, LCDR2 and LCDR3, the LCDR3 of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 25, the LCDR2 of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 24, and the LCDR1 of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 23.
  • the polypeptide of the present application may comprise a first targeting moiety and a second targeting moiety
  • the first targeting moiety may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 of the first targeting moiety may comprise SEQ ID NO:
  • the amino acid sequence shown in 6, the HCDR2 of the first targeting part can include the amino acid sequence shown in SEQ ID NO: 2, and the HCDR1 of the first targeting part can include the amino acid shown in SEQ ID NO: 1 Sequence;
  • the first targeting portion may include LCDR1, LCDR2 and LCDR3, the LCDR3 of the first targeting portion may include the amino acid sequence shown in SEQ ID NO: 25, and the LCDR2 of the first targeting portion may include The amino acid sequence shown in SEQ ID NO: 24, and the LCDR1 of the first targeting part can include the amino acid sequence shown in SEQ ID NO: 23;
  • the second targeting part can include HCDR1, HCDR2 and HCDR3, so
  • the HCDR3 of the second targeting portion may comprise the amino acid
  • the polypeptide of the present application may comprise a first targeting moiety and a second targeting moiety
  • the first targeting moiety may comprise HCDR1, HCDR2 and HCDR3
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 3
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 1
  • the first targeting part may comprise LCDR1, LCDR2 and LCDR3, so
  • the LCDR3 may include the amino acid sequence shown in SEQ ID NO: 25
  • the LCDR2 may include the amino acid sequence shown in SEQ ID NO: 24
  • the LCDR1 may include the amino acid sequence shown in SEQ ID NO: 23
  • the second targeting portion may include HCDR1, HCDR2 and HCDR3, the HCDR3 of the second targeting portion may include the amino acid sequence shown in SEQ ID NO: 14, and the HCDR2 of the second targeting portion may include SEQ ID NO:
  • the LCDR3 of the targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 25
  • the LCDR2 of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 24
  • the LCDR1 of the second targeting portion may Contains the amino acid sequence shown in SEQ ID NO: 23.
  • the polypeptide of the present application can include a first targeting portion and a second targeting portion.
  • the first targeting portion can include HCDR1, HCDR2 and HCDR3.
  • the HCDR3 can include the amino acid sequence shown in SEQ ID NO: 6
  • the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 4
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 1
  • the first targeting part may comprise LCDR1, LCDR2 and LCDR3, so
  • the LCDR3 may include the amino acid sequence shown in SEQ ID NO: 25
  • the LCDR2 may include the amino acid sequence shown in SEQ ID NO: 24
  • the LCDR1 may include the amino acid sequence shown in SEQ ID NO: 23
  • the second targeting portion may include HCDR1, HCDR2 and HCDR3, the HCDR3 of the second targeting portion may include the amino acid sequence shown in SEQ ID NO: 14, and the HCDR2 of the second targeting portion may include SEQ ID NO:
  • the LCDR3 of the targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 25
  • the LCDR2 of the second targeting portion may comprise the amino acid sequence shown in SEQ ID NO: 24
  • the LCDR1 of the second targeting portion may Contains the amino acid sequence shown in SEQ ID NO: 23.
  • the polypeptide of the present application can include a first targeting portion and a second targeting portion, the first targeting portion can include VH and VL, and the VH of the first targeting portion can include SEQ ID NO: 7 to The sequence shown in any one of 11, the VL of the first targeting part may include the sequence shown in SEQ ID NO: 26; the second targeting part may include VH and VL, the second targeting part may Part of the VH may include the sequence shown in SEQ ID NO: 15, and the VL of the second targeting part may include the sequence shown in SEQ ID NO: 26.
  • the first targeting moiety is directly or indirectly linked to the second targeting moiety.
  • the polypeptides have a common light chain.
  • a polypeptide of the present application has multiple antibody light chain variable regions, one or more of which have the same sequence.
  • a polypeptide of the present application has multiple antibody light chains, one or more of which have the same antibody variable region sequence.
  • a polypeptide of the present application has multiple antibody light chains, one or more of which have the same sequence.
  • the polypeptide of the present application has multiple antibody light chains with identical light chain variable region sequences.
  • a polypeptide of the present application has multiple antibody light chains with identical sequences.
  • each heavy chain or light chain amino acid sequence of the antigen-binding protein is homologous to the corresponding amino acid sequence in the antibody from a specific species, or belongs to a specific class.
  • the variable and constant portions of the light and heavy chains are derived from the variable and constant regions of an antibody from one animal species (eg, human).
  • the homolog may be one that shares at least about 85% (e.g., Having at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or higher) sequence homology to a protein or polypeptide.
  • homology generally refers to similarity, similarity or association between two or more sequences. Alignment for the purpose of determining percent sequence homology can be accomplished in a variety of ways known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. One skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms required to achieve maximal alignment within the full-length sequences being compared or within the sequence region of interest. The homology can also be determined by the following methods: FASTA and BLAST. A description of the FASTA algorithm can be found in "Improved Tools for Biological Sequence Comparison" by W.R. Pearson and D.J. Lipman, Proc. Natl.
  • the present application provides a nucleic acid encoding the antigen-binding protein of the present application and/or the polypeptide of the present application.
  • each of the one or more nucleic acid molecules may encode the entire antigen-binding protein or a portion thereof (e.g., HCDR1-3, LCDR1-3, VL, VH, light chain or one or more of the heavy chains).
  • the present application provides a vector, which may comprise the nucleic acid of the present application.
  • a vector which may comprise the nucleic acid of the present application.
  • other genes may be included in the vector, such as marker genes that allow selection of the vector in appropriate host cells and under appropriate conditions.
  • the vector may contain expression control elements that allow correct expression of the coding region in an appropriate host.
  • the vector is an expression vector.
  • the present application provides a cell, which may comprise the antigen-binding protein of the present application, the polypeptide of the present application, and/or the nucleic acid of the present application.
  • each or each host cell may contain one or more nucleic acid molecules or vectors described herein.
  • each or each host cell may comprise multiple (eg, 2 or more) or multiple (eg, 2 or more) nucleic acid molecules or vectors described herein.
  • the present application provides a method for preparing the antigen-binding protein of the present application and/or the polypeptide of the present application, which method includes culturing under conditions that allow the expression of the antigen-binding protein and/or the polypeptide. cells according to the present application. For example, by using appropriate culture medium, appropriate temperature and culture time, etc., these methods are understood by those of ordinary skill in the art.
  • the present application provides a conjugate, which may comprise the antigen-binding protein of the present application and/ Or the polypeptide of the present application.
  • the present application provides a pharmaceutical composition, which may comprise the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cells of the present application, and/ or a conjugate of the present application, and optionally a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable adjuvant is non-toxic to the recipient at the dosage and concentration used.
  • the pharmaceutical composition in the present application may also contain more than one active compound, usually one that does not adversely affect each other's properties. Application active ingredients.
  • the present application provides a kit, which may include the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cells of the present application, and the conjugate of the present application. compounds, and/or pharmaceutical compositions of the present application.
  • the present application provides a method for influencing cells to produce cytokines.
  • the method may comprise administering the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, and the cells of the present application. , the conjugate of the present application, the pharmaceutical composition of the present application, and/or the kit of the present application.
  • the method may be an ex vivo or in vitro method.
  • the method may be a non-therapeutic method.
  • the cells may comprise lymphocytes.
  • T lymphocytes T lymphocytes.
  • the cytokine may comprise an interleukin or/and a functionally active fragment thereof.
  • the cytokine may comprise interleukin 2 (IL-2) or/and functionally active fragments thereof.
  • IL-2 interleukin 2
  • the present application provides a method for promoting cell bridging, which method may comprise administering the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, the cell of the present application, the conjugate of the present application, the The pharmaceutical composition of the application, and/or the kit of the application.
  • the method may be an ex vivo or in vitro method.
  • the method may be a non-therapeutic method.
  • the cell bridging can include reducing the distance between PD-L1 positive cells and PD-1 positive cells.
  • the cell bridging may include increasing the ratio of PD-L1 positive cells and PD-1 positive cells that are double positive, for example, the cell ratio may be detected by flow cytometry.
  • the present application provides an antigen-binding protein of the present application, a polypeptide of the present application, a nucleic acid of the present application, a vector of the present application, a cell of the present application, a conjugate of the present application, and a pharmaceutical composition of the present application. , and/or the use of the kit of the present application in the preparation of medicines for preventing, alleviating and/or treating tumors.
  • the tumor may comprise a PD-L1 high-expressing tumor and/or a PD-L1-positive tumor.
  • the tumor may comprise a solid tumor.
  • the tumor may comprise lung cancer.
  • "PD-L1 high-expressing tumors and/or PD-L1-positive tumors" generally refers to cancers or tumors that contain cancer cells with higher than normal PD-L1 levels.
  • the present application provides an antigen-binding protein of the present application, a polypeptide of the present application, a nucleic acid of the present application,
  • the vector of the present application, the cells of the present application, the conjugate of the present application, the pharmaceutical composition of the present application, and/or the kit of the present application can be used to prevent, alleviate and/or treat tumors.
  • the tumor may comprise a PD-L1 high-expressing tumor and/or a PD-L1-positive tumor.
  • the tumor may comprise a solid tumor.
  • the tumor may comprise lung cancer.
  • the present application provides a method for preventing, alleviating and/or treating tumors, which includes administering the antigen-binding protein of the present application, the polypeptide of the present application, the nucleic acid of the present application, the vector of the present application, and the cells of the present application. , the conjugate of the present application, the pharmaceutical composition of the present application, and/or the kit of the present application.
  • the tumor may comprise a PD-L1 high-expressing tumor and/or a PD-L1-positive tumor.
  • the tumor may comprise a solid tumor.
  • the tumor may comprise lung cancer.
  • sequence information for this application can look like this:
  • a phage antibody single-chain library two rounds of panning were performed against biotin-labeled human PD-L1 (hereinafter referred to as h PD-L1-Biotin, Acro, Catalog No.: PD1-H82E5), and positive enrichment was obtained.
  • the enriched phage was infected with Escherichia coli strain SS320 (purchased from Lucigen) to prepare phage plasmid.
  • Example 2 Yeast display library construction and screening
  • telomere sequence is used as a template to design primers for polymerase chain reaction (PCR) to amplify scFv or VH and VK gene fragments; the PCR-amplified scFv gene fragments are recovered and co-transfected with the yeast display plasmid into the Saccharomyces cerevisiae strain EBY100 (purchased from ATCC), the scFv is inserted into the yeast display plasmid through homologous recombination of Saccharomyces cerevisiae, thereby displaying single-chain antibodies on the surface of the yeast cell wall.
  • PCR polymerase chain reaction
  • the constructed library number is JYYDL168; the PCR amplified VH and VK gene fragments are recovered Then, it was co-transfected into Saccharomyces cerevisiae strain EBY100 with the yeast display plasmids pJYY132-Y and pJYY129-X to construct a yeast display library displaying Fab-form antibodies, library number JYYDL169. After electroporation, the libraries JYYDL168 and JYYDL169 were cultured overnight in 100 mL of SD-Trp medium (Clontech, Cat. No.: 630308) and SD-Trp-Leu (Clontech, Cat.
  • the strength of the combination of Scheme 1 with h PD-L1-Biotin is determined by the PE mean fluorescence signal intensity (MFI) It reflects that the stronger the PE fluorescence intensity, the stronger the binding force between the surface and human PD-L1; similarly, scheme 2 reflects the binding strength of the clone to Cyno PD-L1-Bio, and scheme 3 reflects the binding strength of the clone to irrelevant antigens. , proving the binding specificity of the single clone; Scheme 4 uses Tab2 as a control to perform competitive flow staining to analyze whether the epitopes of each clone and Tab2-binding antigen are the same.
  • MFI PE mean fluorescence signal intensity
  • Example 3 the VH sequences of Y82B4 and Y82F52 clones were selected to construct the IgG1 LALA subtype, and were paired with the light chain plasmid of 41D2HzL4H3 respectively for expression and purification.
  • the candidate antibody numbers were Ab1910T21 and Ab1910T22.
  • the expression results are shown in Table 3 .
  • Octet RED96e (Fortébio) was used to determine the binding of candidate antibodies to human PD-L1 (Acro, Cat. No.: PD1-H5229) and monkey PD-L1 (Acro, Catalog number: PD1-C52H4) affinity.
  • the antigen and antibody were diluted with 1 ⁇ PBST (1 ⁇ PBS: Sangon, B548117-0500; 0.02% Tween 20: sigma, P1379).
  • the concentration of human PD-L1 was 50nM
  • the concentration of monkey PDL1 was 30nM
  • the antibody was used The concentrations are all 33.3nM.
  • Each cycle includes the following steps: 1) Immerse in buffer for 60 seconds; 2) Detect whether the antigen binds non-specifically to the sensor; 3) Regenerate with 10mM glycine solution at pH 1.7; 4) Immerse in buffer for 60 seconds; 5) Antibody solidifies on On the sensor, the time is 23s; 6) The sensor is immersed in the buffer for 180s; 7) The antigen and antibody are combined, the time is 180s; 8) The dissociation of the antigen and antibody, the time is 10 minutes; 9) The sensor is regenerated. For the 1:1 binding mode of antigen-antibody, the association rate (K on ) and dissociation rate (K off ) were measured to calculate the equilibrium dissociation constant (K D ) of the antibody. The results are shown in Table 4 and Table 5 respectively. It can be seen from the table that the antibodies Ab1910T21 and Ab1910T22 of the present application have relatively strong affinity to human or monkey PD-L1.
  • the human Antibody Germline Gene (Data source: IMGT) with the highest homology was selected as the humanization design framework.
  • the heavy chain is framed by IGHV1-2*06 or IGHV1-46*01, IGHJ4*01.
  • the antibody variable regions are numbered according to Chothia rules, the antibody CDR regions are defined according to Chothia, and the amino acids in the antibody heavy chain variable region are humanized based on sequence alignment and variable region structural information.
  • VH sequences T21hzH3 and T22hzH2 were constructed into IgG1 LALA subtypes respectively, which were paired with the light chain plasmid of 41D2HzL4H3 for expression and purification.
  • the candidate antibody numbers and expression results are shown in Table 6.
  • Example 8 It can be seen from Example 8 that the humanized antibodies 2005K4T21hz3 and 2005K4T22hz2 have strong affinity with human or monkey PD-L1.
  • Example 5 we further evaluated the ability of the antibodies before and after humanization and the reference antibody Atezolizumab to activate mixed lymphocytes to release IL-2. Activity, the results are shown in Figures 2 and 3.
  • Figure 2 shows that Ab1910T22 humanized antibody 2005K4T22hz2 still has the function of enhancing T cell activation and releasing IL-2 in vitro after humanization
  • Figure 3 shows that Ab1910T21 humanized antibody 2005K4T21hz3 still has the function of enhancing T cells in vitro after humanization.
  • the function of T cell activation and release of IL-2, among which the function of 2005K4T21hz3 is better than that of the benchmark antibody Atezolizumab.
  • a co-light chain double antibody was constructed using the 2005K4T21hz3 and 2005K4T22hz2 humanized antibody VH sequences and the 41D2HzL4H3 antibody sequence.
  • IgG1 LALA D265S and knob and Hole designs were selected, as shown in Figure 4; among them, the Hole of BAb2005.05
  • the CDR2 in the sequence contains the PTM site of NS, which was mutated into NY, and the antibody was named BAb2005.05.2. All antibodies were expressed and purified.
  • the antibody numbers and expression results are shown in Table 9.
  • Table 11 shows that the affinity of BAb2005.04, BAb2005.05, and BAb2005.05.2 to monkey PDL1 is equivalent to the parent PD-L1 monoclonal antibody Ab2005-H.1 and stronger than the control antibody Atezolizumab.
  • Table 12 shows that the affinity of BAb2005.04, BAb2005.05, and BAb2005.05.2 to human PD1 is equivalent to the parent PD-1 monoclonal antibody Ab2005-K and stronger than the control antibody Pembrolizumab.
  • Table 13 shows that the affinity of BAb2005.04, BAb2005.05, and BAb2005.05.2 to monkey PD1 is equivalent to the parent PD-1 monoclonal antibody Ab2005-K, but weaker than the control antibody Pembrolizumab.
  • Co-light chain bispecific antibodies can stimulate mixed lymphocytes to release IL-2
  • Example 5 further evaluate the co-light chain bispecific antibodies BAb2005.04, BAb2005.05, BAb2005.05.2 and the maternal PD-L1 monoclonal antibody Ab2005-H.1 and the maternal PD-1 monoclonal antibody Ab2005-K, And the PD-L1 control antibody Atezolizumab and PD-1 control antibody Pembrolizumab activate the activity of mixed lymphocytes to release IL-2.
  • PD-H.1 and the parent PD-1 monoclonal antibody Ab2005-K have the function of enhancing T cell activation and releasing IL-2 in vitro, and their functions are better than the PD-L1 control antibody Atezolizumab and the PD-1 control antibody Pembrolizumab.
  • Example 13 Co-light chain bispecific antibody has cell bridging function
  • the cell bridging function method was used to evaluate the co-light chain bispecific antibodies BAb2005.04, BAb2005.05, BAb2005.05.2 and the maternal PD-L1 monoclonal antibody Ab2005-H.1 and the maternal PD-1 monoclonal antibody Ab2005-K.
  • the specific steps are as follows:
  • CHOK1-PD-1 was stained with 3 ⁇ M CFSE dye (Invitrogen, Cat. No.: C34554), and CHOK1-PD-L1 cells were stained with 1 ⁇ M CellTrace TM Red dye (Invitrogen, Cat. No.: C34564).
  • the specific staining method is: The cells were centrifuged at 400 g for 10 min, and the cell pellet was resuspended in suspension (PBS+2% FBS). Then filter through a 40 ⁇ m mesh, adjust the cell density to 1x 10 7 cells/mL, add an equal volume of the corresponding 2-fold concentration dye, and mix immediately upon addition.
  • the highest final concentration of the antibody is 22.5 ⁇ g/mL (prepared concentration is 45 ⁇ g/mL), 3-fold gradient dilution (11 concentration points + 1 0 concentration point), and simultaneously dilute the parent PD- 1.
  • the final concentration of monoclonal antibody Ab2005-K and negative control antibody anti-HEL Human IgG1 is 90 ⁇ g/mL (prepared concentration is 180 ⁇ g/mL).
  • the prepared cells and antibodies in equal volumes i.e., 60 ⁇ L cells + 60 ⁇ L antibodies
  • the wells on the PD-L1 side are CHOK1-PD-L1+ serially diluted co-light chain bispecific antibodies
  • the control wells are CHOK1-PD-L1+ gradiently diluted parent monoclonal antibody combination.
  • the prepared cells and antibodies to the corresponding cell culture plate as 60 ⁇ L cells + 150 ⁇ L antibody + 90 ⁇ L medium.
  • the specific setting is CHOK1-PD-1+Ab2005-K/anti-HEL Human IgG1. Cells and antibodies were added to the cell culture plate, mixed and incubated at 4°C for 2 hours.
  • adding the maternal PD-L1 monoclonal antibody Ab2005-H.1 and the maternal PD-1 monoclonal antibody Ab2005-K at the same time does not have a cell bridging function, and adding the maternal PD-1 monoclonal antibody Ab2005.05 at the same time Anti-Ab2005-K can block the cell bridging function of the double antibody.
  • NPG mice were purchased from Beijing Weitongda Animal Breeding Co., Ltd., 5 weeks old, female, 56 in total.
  • Frozen PBMC were purchased from Miaoshun (Shanghai) Biotechnology Co., Ltd.
  • NCI-H292 human lung cancer cells were purchased from ATCC and cultured in RPMI-1640 medium supplemented with 10% FBS in a 37°C incubator containing CO2 . Before the cells were continuously cultured for ten generations, approximately 4 ⁇ 10 6 NCI-H292 cells were suspended in 50 ⁇ L PBS and an equal volume of 1 ⁇ 10 6 PBMC cells were mixed, and then mixed with an equal volume of Matrigel, and inoculated into the The inoculation volume was 200 ⁇ L per mouse.
  • mice On the day of inoculation, 56 mice were randomly divided into 7 groups according to their weight, with 8 mice in each group. The day of group administration was defined as day 0.
  • Group G1 was administered with isotype control, group G2 was administered with Ab2005-K at a dose of 5 mg/kg; group G3 was administered with Ab2005-H.1 at a dose of 5 mg/kg; group G4 was administered with Ab2005-K combined with Ab2005-H.1 at a dose of 5mg/kg+5mg/kg; G5 group was administered BAb2005.05, dose 5mg/kg; G6 group was administered BAb2005.05.2, dose 5mg/kg; G7 group was administered BAb2005.04, dose 5mg/kg. All administration methods were intraperitoneal injection, twice a week, for a total of 6 administrations.
  • the average tumor volume of group G1 was 688.30 ⁇ 50.11mm 3 and that of group G2
  • the average tumor volume of the G3 group was 6.00 ⁇ 6.00mm 3
  • the average tumor volume of the G3 group was 8.55 ⁇ 8.55mm 3
  • the average tumor volume of the G4 group was 13.65 ⁇ 6.61mm 3
  • the average tumor volume of the G5 group was 6.16 ⁇ 4.20mm 3
  • the average tumor volume of the G6 group was 6.00 ⁇ 6.00mm 3 .
  • the tumor volume was 5.80 ⁇ 5.80mm 3
  • the average tumor volume in the G7 group was 0.00 ⁇ 0.00mm 3 .
  • the G2 group has a tumor inhibition rate of 101.45%
  • the G3 group has a tumor inhibition rate of 101.45%
  • the G4 group has a tumor inhibition rate of 100.87%
  • the G5 group has a tumor inhibition rate of 101.69%
  • the G6 group has a tumor inhibition rate of 101.21%.
  • the G7 group had a tumor inhibition rate of 101.50%.
  • the G2 and G3 groups had extremely significant statistical differences compared with the G1 group, G2 vs G1, P ⁇ 0.001; G3 vs G1, P ⁇ 0.001. It was proved that both Ab2005-K and Ab2005-H.1 single drugs can significantly inhibit tumor growth. There was a very significant statistical difference between the G4 group and the G1 group, G4 vs G1, P ⁇ 0.001.

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Abstract

La présente invention concerne une protéine de liaison à l'antigène, et concerne spécifiquement une protéine de liaison à PD-L1 et/ou concerne une protéine de liaison bispécifique capable de se lier à PD-L1 et à PD-1. La présente invention concerne en outre une application de la protéine de liaison à l'antigène dans le traitement de maladies ou de troubles.
PCT/CN2023/086611 2022-04-08 2023-04-06 Protéine de liaison à l'antigène et son application WO2023193767A1 (fr)

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Publication number Priority date Publication date Assignee Title
CN113166269A (zh) * 2018-11-13 2021-07-23 指南针制药有限责任公司 对抗检查点分子的多特异性结合构建体及其用途
WO2021226984A1 (fr) * 2020-05-15 2021-11-18 三生国健药业（上海）股份有限公司 Anticorps bispécifique tétravalent contre pd-1 et pd-l1

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113166269A (zh) * 2018-11-13 2021-07-23 指南针制药有限责任公司 对抗检查点分子的多特异性结合构建体及其用途
WO2021226984A1 (fr) * 2020-05-15 2021-11-18 三生国健药业（上海）股份有限公司 Anticorps bispécifique tétravalent contre pd-1 et pd-l1

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