WO2022223004A1 - Anticorps anti-siglec-15 et son utilisation - Google Patents

Anticorps anti-siglec-15 et son utilisation Download PDF

Info

Publication number
WO2022223004A1
WO2022223004A1 PCT/CN2022/088241 CN2022088241W WO2022223004A1 WO 2022223004 A1 WO2022223004 A1 WO 2022223004A1 CN 2022088241 W CN2022088241 W CN 2022088241W WO 2022223004 A1 WO2022223004 A1 WO 2022223004A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
amino acid
acid sequence
binding protein
antibody
Prior art date
Application number
PCT/CN2022/088241
Other languages
English (en)
Chinese (zh)
Inventor
廖雪梅
栾珊珊
杨松霖
陈娜
施伟军
王瓅
刘颖颖
李贵祥
刘婷婷
王小迪
夏广新
柯樱
Original Assignee
上海医药集团股份有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 上海医药集团股份有限公司 filed Critical 上海医药集团股份有限公司
Priority to CN202280029885.0A priority Critical patent/CN117203241A/zh
Publication of WO2022223004A1 publication Critical patent/WO2022223004A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • 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
    • A61P19/00—Drugs for skeletal disorders
    • 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
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor

Definitions

  • the application provides an isolated antigen-binding protein, which has one or more of the following properties: 1) in Biacore detection, specifically binds to human Siglec15 protein with a KD value of about 4E-09M or less; 2) In flow assay, it specifically binds to human Siglec15 expressed on the surface of CHOK1 cells with an EC50 value of about 0.1 ⁇ g/ml or less; 3) Relieves the proliferation inhibition of PBMC in blood by Siglec15; 4) In ELISA assay , specifically binds to human Siglec15 with an EC50 value of about 0.2 ⁇ g/ml or less; 5) specifically binds to monkey Siglec15 with an EC50 value of about 0.1 ⁇ g/ml or less in an ELISA assay; and 6) in an ELISA assay , specifically binds to murine Siglec15 with an EC50 value of about 0.1 ⁇ g/ml or less.
  • the isolated antigen binding protein comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:4.
  • the H-FR1, H-FR2, H-FR3 and H-FR4 in the isolated antigen binding protein comprise an amino acid sequence selected from any of the following groups:
  • the isolated antigen binding protein comprises a heavy chain variable region VH comprising the amino acid sequence set forth in SEQ ID NO:72.
  • the VH in the isolated antigen binding protein comprises the amino acid sequence set forth in any one of SEQ ID NOs: 29-32.
  • the isolated antigen binding protein comprises L-FR1, the C-terminus of L-FR1 is directly or indirectly linked to the N-terminus of LCDR1, and the L-FR1 comprises SEQ ID NO : amino acid sequence shown in 68.
  • the L-FR1 in the isolated antigen binding protein comprises the amino acid sequence set forth in any one of SEQ ID NOs: 16-18.
  • the isolated antigen-binding protein comprises L-FR2, the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 comprises SEQ ID NO: 69 amino acid sequence.
  • the L-FR3 in the isolated antigen binding protein comprises the amino acid sequence set forth in any one of SEQ ID NOs: 23-25.
  • the L-FR4 in the isolated antigen binding protein comprises the amino acid sequence set forth in any one of SEQ ID NOs: 27 and 28.
  • the isolated antigen binding protein comprises L-FR1, L-FR2, L-FR3 and L-FR4, the L-FR1 comprising the amino acid sequence set forth in SEQ ID NO: 68; the L-FR2 comprises the amino acid sequence shown in SEQ ID NO:69; the L-FR3 comprises the amino acid sequence shown in SEQ ID NO:70; and the L-FR4 comprises the amino acid sequence shown in SEQ ID NO:71.
  • the L-FR1, L-FR2, L-FR3 and L-FR4 in the isolated antigen binding protein comprise an amino acid sequence selected from any of the following groups:
  • L-FR1 SEQ ID NO: 16
  • L-FR2 SEQ ID NO: 20
  • L-FR3 SEQ ID NO: 23
  • L-FR4 SEQ ID NO: 27;
  • L-FR1 SEQ ID NO: 17, L-FR2: SEQ ID NO: 21, L-FR3: SEQ ID NO: 24 and L-FR4: SEQ ID NO: 28;
  • L-FR1 SEQ ID NO: 18, L-FR2: SEQ ID NO: 21, L-FR3: SEQ ID NO: 25 and L-FR4: SEQ ID NO: 28;
  • L-FR1 SEQ ID NO: 18, L-FR2: SEQ ID NO: 20, L-FR3: SEQ ID NO: 25 and L-FR4: SEQ ID NO: 28.
  • the isolated antigen binding protein comprises a light chain variable region VL comprising the amino acid sequence set forth in SEQ ID NO:73.
  • the VL in the isolated antigen binding protein comprises the amino acid sequence set forth in any one of SEQ ID NOs: 33-36.
  • the VH and VL in the isolated antigen binding protein comprise amino acid sequences selected from any of the following groups:
  • VH SEQ ID NO: 29 and VL: SEQ ID NO: 33;
  • VH SEQ ID NO:30 and VL: SEQ ID NO:36;
  • VH SEQ ID NO:31 and VL: SEQ ID NO:35;
  • VH SEQ ID NO: 31 and VL: SEQ ID NO: 36;
  • VH SEQ ID NO: 32 and VL: SEQ ID NO: 34;
  • VH SEQ ID NO:32 and VL: SEQ ID NO:35;
  • VH SEQ ID NO:32 and VL: SEQ ID NO:36.
  • the isolated antigen binding protein comprises a heavy chain constant region, and the heavy chain constant region comprises an IgG-derived constant region or an IgY-derived constant region.
  • the heavy chain constant region in the isolated antibody binding protein comprises a constant region derived from human IgG.
  • the light chain constant region in the isolated antigen binding protein comprises a constant region derived from human Ig ⁇ .
  • the light chain constant region in the isolated antigen binding protein comprises the amino acid sequence set forth in any one of SEQ ID NO:43.
  • the isolated antigen-binding protein comprises an antibody or antigen-binding fragment thereof.
  • the antigen-binding fragment in the isolated antigen-binding protein is selected from the group consisting of Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di- scFv, VHH and/or dAb.
  • the antibody in the isolated antigen binding protein is selected from the group consisting of monoclonal antibodies, single chain antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.
  • the application provides a polypeptide comprising the isolated antigen binding protein.
  • the application provides an immunoconjugate comprising the isolated antigen binding protein or the polypeptide.
  • the application provides an isolated nucleic acid molecule encoding the isolated antigen binding protein, or the polypeptide.
  • the application provides a cell comprising the isolated antigen binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule and/or the carrier.
  • the present application provides a pharmaceutical composition
  • a pharmaceutical composition comprising the isolated antigen binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell, and/or pharmaceutically acceptable adjuvants and/or excipients.
  • the application provides a pharmaceutical combination comprising the isolated antigen binding protein and an immune checkpoint inhibitor.
  • the immune checkpoint inhibitor includes a substance that inhibits the PD-1/PD-L1 interaction.
  • the immune checkpoint inhibitor is selected from the group consisting of PD-1/PD-L1 blockers, PD-1 antagonists, PD-L1 antagonists, PD-1 inhibitors and PD-1 L1 inhibitor.
  • the immune checkpoint inhibitor comprises an anti-PD-1 antibody.
  • the anti-PD-1 antibody comprises a HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:46.
  • the anti-PD-1 antibody comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:45.
  • the anti-PD-1 antibody comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:44.
  • the anti-PD-1 antibody comprises a heavy chain variable region VH
  • the VH comprises HCDR1, HCDR2 and HCDR3
  • the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 46
  • the HCDR2 comprise the amino acid sequence shown in SEQ ID NO:45
  • the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:44.
  • the anti-PD-1 antibody comprises a heavy chain variable region VH comprising the amino acid sequence set forth in SEQ ID NO:50.
  • the anti-PD-1 antibody comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:49.
  • the anti-PD-1 antibody comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:48.
  • the anti-PD-1 antibody comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:47.
  • the anti-PD-1 antibody comprises a light chain variable region VL
  • the VL comprises LCDR1, LCDR2 and LCDR3
  • the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 49
  • the LCDR2 comprise the amino acid sequence shown in SEQ ID NO:48
  • the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:47.
  • the anti-PD-1 antibody comprises a light chain variable region VL comprising the amino acid sequence set forth in SEQ ID NO:51.
  • the anti-PD-1 antibody comprises pembrolizumab.
  • the immune checkpoint inhibitor comprises an anti-PD-L1 antibody.
  • the anti-PD-L1 antibody comprises HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:60.
  • the anti-PD-L1 antibody comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:59.
  • the anti-PD-L1 antibody comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:58.
  • the anti-PD-L1 antibody comprises a heavy chain variable region VH
  • the VH comprises HCDR1, HCDR2 and HCDR3
  • the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 60
  • the HCDR2 comprise the amino acid sequence shown in SEQ ID NO:59
  • the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:58.
  • the anti-PD-L1 antibody comprises a heavy chain variable region VH comprising the amino acid sequence set forth in SEQ ID NO:54.
  • the anti-PD-L1 antibody comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:63.
  • the anti-PD-L1 antibody comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:62.
  • the anti-PD-L1 antibody comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:61.
  • the anti-PD-L1 antibody comprises a light chain variable region VL
  • the VL comprises LCDR1, LCDR2 and LCDR3
  • the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 63
  • the LCDR2 comprise the amino acid sequence shown in SEQ ID NO:62
  • the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:61.
  • the anti-PD-L1 antibody comprises atezolizumab.
  • the pharmaceutical combination may be a pharmaceutical composition.
  • the present application provides a method for detecting or assaying Siglec15, the method comprising using the isolated antigen binding protein or the polypeptide.
  • the present application provides a Siglec15 detection kit, which comprises the isolated antigen-binding protein or the polypeptide.
  • the present application provides a use of the isolated antigen-binding protein or the polypeptide in the preparation of a kit.
  • the application provides a method of modulating an immune response, comprising administering to a subject in need thereof an effective amount of the isolated antigen binding protein, the polypeptide, the immunoconjugate, the The isolated nucleic acid molecule, the vector, the cell and/or the pharmaceutical composition, and/or the pharmaceutically acceptable therapeutic agent.
  • the application provides a method of modulating an immune response comprising administering to a subject in need thereof an effective amount of the pharmaceutical combination, and/or a pharmaceutically acceptable therapeutic agent.
  • the application provides the isolated antigen binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell and/or the pharmaceutical composition, It is used to prevent, alleviate and/or treat a disease or disorder.
  • the application provides the pharmaceutical combination for the prevention, alleviation and/or treatment of a disease or disorder.
  • the disease or disorder comprises abnormal bone metabolism.
  • the abnormal bone metabolism comprises osteoporosis, bone destruction associated with rheumatoid arthritis, cancerous hypercalcemia, bone destruction associated with multiple myeloma or bone metastases from cancer, bone destruction Loss of tooth mass due to periodontitis, osteolysis around artificial joints, bone destruction in chronic osteomyelitis, Paget's disease of bone, renal osteodystrophy, and osteogenesis imperfecta.
  • the abnormal bone metabolism comprises osteoporosis.
  • the disease or disorder comprises a tumor.
  • the tumor comprises a tumor associated with the expression of Siglec15.
  • the tumor comprises a solid tumor.
  • the tumor comprises colon cancer.
  • the present application provides the isolated antigen binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell and/or the drug combination Use of the substance in the preparation of a medicament for the prevention and/or treatment of a disease or disorder.
  • the present application provides the use of the pharmaceutical combination in the manufacture of a medicament for preventing and/or treating a disease or disorder.
  • the disease or disorder comprises abnormal bone metabolism.
  • the abnormal bone metabolism comprises osteoporosis, bone destruction associated with rheumatoid arthritis, cancerous hypercalcemia, bone destruction associated with multiple myeloma or bone metastases from cancer, bone destruction Loss of tooth mass due to periodontitis, osteolysis around artificial joints, bone destruction in chronic osteomyelitis, Paget's disease of bone, renal osteodystrophy, and osteogenesis imperfecta.
  • the abnormal bone metabolism comprises osteoporosis.
  • the disease or disorder comprises a tumor.
  • the tumor comprises a tumor associated with the expression of Siglec15.
  • the tumor comprises a solid tumor.
  • the tumor comprises colon cancer.
  • the present application provides a method of preventing and/or treating a disease or disorder, comprising administering to a subject in need thereof an effective amount of the isolated antigen binding protein, the polypeptide, the immunoconjugate compound, the isolated nucleic acid molecule, the vector, and/or the cell.
  • the present application provides a method of preventing and/or treating a disease or disorder comprising administering to a subject in need thereof an effective amount of the pharmaceutical combination.
  • the disease or disorder comprises abnormal bone metabolism.
  • the abnormal bone metabolism comprises osteoporosis, bone destruction associated with rheumatoid arthritis, cancerous hypercalcemia, bone destruction associated with multiple myeloma or bone metastases from cancer, bone destruction Decreased quality of life, tooth loss due to periodontitis, osteolysis around artificial joints, bone destruction in chronic osteomyelitis, Paget's disease of bone, renal osteodystrophy, and osteogenesis imperfecta.
  • the abnormal bone metabolism comprises osteoporosis.
  • the disease or disorder comprises a tumor.
  • the tumor comprises a tumor associated with the expression of Siglec15.
  • the tumor comprises a solid tumor.
  • the tumor comprises colon cancer.
  • Figure 1 shows the husiglec-15-hFc antigenic activity measured by the ELISA described herein.
  • Figure 2A shows the cyno-siglec-15-his antigenic activity as measured by the ELISA described herein.
  • Figure 2B shows the husiglec-15-his antigenic activity measured by the ELISA described herein.
  • Figure 3 shows serum titers from multiple immunized mice.
  • Figure 4 shows the binding curve of an exemplary Siglec15 antigen binding protein to human Siglec15 expressed on the surface of CHOK1 cells.
  • Figure 5A shows the effect of the exemplary Siglec15 antigen binding protein described herein on CD8+ T cell proliferation.
  • Figure 5B shows the effect of the exemplary Siglec15 antigen binding protein described herein on CD4+ T cell proliferation.
  • Figure 5C shows the effect of the exemplary Siglec15 antigen binding protein described herein on IFN- ⁇ expression.
  • Figure 6A shows the binding curve of an exemplary humanized Siglec15 binding protein described herein to human Siglec15 expressed on the surface of CHOK1 cells.
  • Figure 6B shows the binding curve of the exemplary humanized Siglec15 binding protein described herein to human Siglec15 expressed on the surface of CHOK1 cells.
  • Figure 7A shows the effect of the exemplary humanized Siglec15 antigen binding protein described herein on CD8+ T cell proliferation.
  • Figure 7B shows the effect of the exemplary humanized Siglec15 antigen binding protein described herein on CD4+ T cell proliferation.
  • Figure 7C shows the effect of the exemplary humanized Siglec15 antigen binding protein described herein on IFN- ⁇ expression.
  • Figure 8A shows the binding curve of an exemplary humanized Siglec15 antigen binding protein described herein to human Siglec15 antigen.
  • Figure 8B shows the binding curve of the exemplary humanized Siglec15 antigen binding protein described herein to the monkey Siglec15 antigen.
  • Figure 8C shows the binding curve of an exemplary humanized Siglec15 antigen binding protein described herein to murine Siglec15 antigen.
  • Figure 9A shows the results of the anti-tumor efficacy test of the exemplary anti-Siglec15 antibody Ab0 described in the present application in wild-type C57BL/6N mice.
  • Figure 9B shows body weight changes in wild-type C57BL/6N mice following administration of an exemplary anti-Siglec15 antibody Ab0.
  • Figure 10A shows the results of the anti-tumor efficacy test of the exemplary anti-Siglec15 antibodies Abl, Ab6 and Ab7 described in this application in wild-type C57BL/6N mice.
  • Figure 10B shows body weight changes in wild-type C57BL/6N mice following administration of exemplary anti-Siglec15 antibodies Ab1, Ab6 and Ab7.
  • Figure 11A shows the results of the anti-tumor efficacy test of the exemplary anti-Siglec15 antibodies Ab0, Ab1, Ab6 and Ab7 described in the present application in human Siglec15 transgenic mice.
  • Figure 11B shows body weight changes in human Siglec15 transgenic mice following administration of exemplary anti-Siglec15 antibodies AbO, Ab1, Ab6 and Ab7.
  • Figure 12 shows that humanized Siglec15 antigen binding protein inhibits osteoclast formation.
  • Figure 13A shows the anti-tumor efficacy of the exemplary Siglec15 antigen-binding proteins Ab1, Ab7 and PD-1 antibodies in combination in human PD-1/PD-L1 transgenic mice.
  • Figure 13B shows the body weight change of human PD-1/PD-L1 transgenic mice after the exemplary Siglec15 antigen-binding protein Ab1, Ab7 and PD-1 antibody are used in combination.
  • Figure 14A shows the anti-tumor efficacy of the exemplary Siglec15 antigen-binding proteins Ab1, Ab7 and PD-L1 antibodies in combination in human PD-1/PD-L1 transgenic mice.
  • Figure 14B shows the body weight change of human PD-1/PD-L1 transgenic mice after the combination of the exemplary Siglec15 antigen binding proteins Ab1, Ab7 and PD-L1 antibody described in the present application.
  • isolated generally refers to artificial means obtained from the natural state. If an "isolated" substance or component occurs in nature, it may be due to a change in its natural environment, or separation of the substance from its natural environment, or both. For example, a certain unisolated polynucleotide or polypeptide naturally exists in a living animal, and the same polynucleotide or polypeptide with high purity isolated from this natural state is called isolated of.
  • isolated does not exclude the admixture of artificial or synthetic materials, nor does it exclude the presence of other impurities that do not affect the activity of the material.
  • the term “antigen binding protein” generally refers to a polypeptide molecule capable of specifically recognizing and/or neutralizing a specific antigen.
  • the term “antigen-binding protein” may include “antibody” or "antigen-binding fragment”.
  • the antibody may comprise an immunoglobulin consisting of at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, and may comprise any molecule comprising an antigen-binding portion thereof.
  • antibody may include monoclonal antibodies, antibody fragments or antibody derivatives including, but not limited to, murine antibodies, human antibodies (fully human antibodies), humanized antibodies, chimeric antibodies, single chain antibodies (eg, scFv ), as well as antigen-binding antibody fragments (eg, Fab, Fab', VHH and (Fab)2 fragments).
  • antibody may also include all recombinant forms of antibodies, such as antibodies expressed in prokaryotic cells, unglycosylated antibodies, and any antigen-binding antibody fragments and derivatives thereof described herein.
  • Each heavy chain can be composed of a heavy chain variable region (VH) and a heavy chain constant region.
  • Each light chain can be composed of a light chain variable region (VL) and a light chain constant region.
  • VH and VL regions can be further distinguished into hypervariable regions called complementarity determining regions (CDRs) interspersed in more conserved regions called framework regions (FRs).
  • CDRs complementarity determining regions
  • FRs framework regions
  • Each VH and VL can consist of three CDRs and four FR regions, which can be arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.
  • the variable regions of the heavy and light chains contain binding domains that interact with antigen (eg, human Siglec15).
  • the constant region of the antibody mediates the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (Clq) of the classical complement system.
  • the exact boundaries of the CDRs have been defined differently from system to system.
  • the system described by Kabat Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) provides not only a The residue numbering system is specified, and precise residue boundaries are provided that define the CDRs. These CDRs may be referred to as Kabat CDRs. Chothia and colleagues (Chothia and Lesk, J. Mol. Biol.
  • CDRs Boundary definitions may not strictly follow one of the above systems, but will still overlap with Kabat CDRs, although they can be shortened or lengthened according to predictions or experimental findings that specific residues or groups of residues or even entire CDRs do not significantly affect antigen binding
  • the CDRs can be defined by using the Kabat numbering system.
  • the term "antigen-binding fragment” generally refers to one or more fragments of an antibody that function to specifically bind an antigen.
  • the antigen-binding function of an antibody can be achieved by full-length fragments of the antibody.
  • Antigen binding function of an antibody can also be achieved by a heavy chain comprising a fragment of Fv, ScFv, dsFv, Fab, Fab' or F(ab')2, or alternatively, comprising Fv, scFv, dsFv, Fab, Fab' or The light chain of a fragment of F(ab')2.
  • Fab fragment usually a monovalent fragment consisting of VL, VH, CL and CH domains;
  • F(ab')2 fragment comprising two Fab fragments linked by a disulfide bond at the hinge region (3) Fd fragment composed of VH and CH domains; (4) Fv fragment composed of VL and VH domains of antibody one-arm; (5) dAb fragment composed of VH domain (Ward et al, (1989) Nature 341:544-546); (6) a combination of separate complementarity determining regions (CDRs) and (7) two or more separate CDRs optionally linked by a linker.
  • CDRs complementarity determining regions
  • a monovalent single-chain molecule Fv (scFv) formed by the pairing of VL and VH may also be included (see Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. 85 : 5879-5883).
  • a class of antibody VHHs that lack the antibody light chain and only have the heavy chain variable region can also be included (for example, see Kang Xiaozhen et al., Chinese Journal of Biological Engineering, 2018, 34(12): 1974-1984).
  • the "antigen-binding portion” may also include an immunoglobulin fusion protein comprising a binding domain selected from the group consisting of: (1) a binding domain polypeptide fused to an immunoglobulin hinge region polypeptide; (2) with an immunoglobulin heavy chain CH2 constant region fused to the hinge region; and (3) an immunoglobulin heavy chain CH3 constant region fused to the CH2 constant region.
  • an immunoglobulin fusion protein comprising a binding domain selected from the group consisting of: (1) a binding domain polypeptide fused to an immunoglobulin hinge region polypeptide; (2) with an immunoglobulin heavy chain CH2 constant region fused to the hinge region; and (3) an immunoglobulin heavy chain CH3 constant region fused to the CH2 constant region.
  • the term "monoclonal antibody” generally refers to a population of substantially homologous antibodies, ie the individual antibodies comprising the population are identical except for possible naturally occurring mutations present in minor amounts.
  • Monoclonal antibodies are highly specific, directed against a single antigenic site.
  • the monoclonal antibodies can be prepared by hybridoma technology or produced in bacterial, eukaryotic, or plant cells by using recombinant DNA methods.
  • Monoclonal antibodies can also be obtained from phage antibody libraries, using techniques such as those described by Clackson et al., Nature, 352:624-628 (1991) and Marks et al., Mol. Biol., 222:581-597 (1991). conduct.
  • chimeric antibody generally refers to an antibody in which a portion of each heavy or light chain amino acid sequence is homologous to the corresponding amino acid sequence in an antibody from a particular species, or belongs to a particular class, while The remainder of the chain is then homologous to the corresponding sequence in another species.
  • variable regions of both light and heavy chains are derived from variable regions of antibodies from one animal species (eg, mouse, rat, etc.), while the constant portions are homologous to antibody sequences from another species (eg, human) .
  • non-human B cells or hybridoma cells can be used to generate variable regions, and the constant regions combined therewith are derived from humans.
  • variable regions have the advantage of being easy to prepare and their specificity is not affected by the source of the constant regions with which they are combined.
  • the constant region of the chimeric antibody can be derived from humans, the possibility of eliciting an immune response when the chimeric antibody is injected is lower than that of using an antibody whose constant region is of non-human origin.
  • humanized antibody generally refers to a chimeric antibody that contains less sequence from non-human immunoglobulins, thereby reducing the immunogenicity of the xenogeneic antibody when introduced into humans, while at the same time The full antigen-binding affinity and specificity of the antibody is maintained.
  • CDR grafting (Jones et al., Nature 321:522 (1986)) and variants thereof can be used; including “reshaping”, (Verhoeyen, et al., 1988 Science 239:1534-1536; Riechmann, et al., 1988 Nature 332:323-337; Tempest, et al., Bio/Technol 1991 9:266-271), "hyperchimerization", (Queen, et al., 1989 Proc Natl Acad Sci USA 86 : 10029-10033; Co, et al., 1991 Proc Natl Acad Sci USA 88: 2869-2873; Co, et al., 1992 J Immunol 148: 1149-1154) and "veneering", (Mark, et al ., "Derivation of therapeutically active humanized and veneered anti-CD18antibodies.” In: Metcalf B W, Dalton B J, eds.
  • murine antibody generally refers to antibodies in which the variable region framework and CDR regions are derived from mouse germline immunoglobulin sequences. Furthermore, if the antibody contains constant regions, it is also derived from mouse germline immunoglobulin sequences.
  • the murine antibodies of the present application may comprise amino acid residues not encoded by mouse germline immunoglobulin sequences, eg, may include mutations introduced by random or point mutation in vitro or by somatic mutation in vivo.
  • Siglec15 protein Siglec-15
  • Siglec15 antigen any functionally active fragments, variants and homologues of Siglec15 that are naturally expressed by cells or are in use Siglec15 gene transfected cells expressed.
  • Siglec15 may be human Siglec15, whose accession number in UniProt/Swiss-Prot is Q6ZMC9.
  • Siglec15 can be a functionally active fragment of human Siglec15.
  • the “functionally active fragment” can include a fragment that retains the endogenous function of at least one naturally occurring protein (eg, binds to the antigen binding proteins described herein).
  • the "functionally active fragment” may include a domain that binds to the antigen-binding protein of the present application.
  • the present application may also include functionally active fragments, derivatives, analogs, homologues, and fragments thereof.
  • a functionally active fragment refers to a polypeptide having substantially the same amino acid sequence or encoded by substantially the same nucleotide sequence as the naturally-occurring sequence and capable of possessing one or more activities of the naturally-occurring sequence.
  • a functionally active fragment of any given sequence refers to one in which a particular sequence of residues (whether amino acid or nucleotide residues) has been modified such that the polypeptide or polynucleotide substantially retains at least at least A sequence of endogenous functions.
  • Sequences encoding functionally active fragments can be obtained by addition, deletion, substitution, modification, substitution and/or variation of at least one amino acid residue and/or nucleotide residue present in a naturally occurring protein and/or polynucleotide , as long as the original functional activity is maintained.
  • the term "derivative" generally refers to the polypeptide or polynucleotide of the present application including any substitution, variation, modification, substitution, deletion and /or addition, so long as the resulting polypeptide or polynucleotide substantially retains at least one of its endogenous functions.
  • analog generally refers to a polypeptide or polynucleotide and includes any mimetic of the polypeptide or polynucleotide, ie possessing at least one endogenous function of the polypeptide or polynucleotide that the mimetic mimics chemical compounds.
  • amino acid substitutions such as at least 1 (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) amino acid substitutions, can be made, so long as the modified sequence remains substantially as desired activity or ability.
  • Amino acid substitutions can include the use of non-naturally occurring analogs.
  • homologue generally refers to an amino acid sequence or nucleotide sequence that has some homology to a naturally occurring sequence.
  • the term “homology” may be equivalent to sequence "identity”.
  • homologous sequences can include amino acid sequences that can 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 .
  • a homologue will contain the same active site, etc., as the subject amino acid sequence.
  • Homology can be considered in terms of similarity (ie, amino acid residues with similar chemical properties/functions), or it can be expressed in terms of sequence identity.
  • a reference to a sequence having a percent identity to any one of the SEQ ID NOs of an amino acid sequence or a nucleotide sequence refers to that percent identity over the entire length of the referenced SEQ ID NO. the sequence of.
  • sequence alignments can be performed by various means known to those skilled in the art, eg, using BLAST, BLAST-2, ALIGN, NEEDLE or Megalign (DNASTAR) software and the like. Those skilled in the art can determine appropriate parameters for alignment, including any algorithms needed to achieve optimal alignment among the full-length sequences being compared.
  • proteins or polypeptides used in the present application may also have deletions, insertions or substitutions of amino acid residues that produce silent changes and result in functionally equivalent proteins.
  • Deliberate amino acid substitutions can be made based on similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or amphiphilic properties of the residues, so long as endogenous function is preserved.
  • negatively charged amino acids include aspartic acid and glutamic acid
  • positively charged amino acids include lysine and arginine
  • amino acids containing uncharged polar headgroups with similar hydrophilicity values include amino acids Paraparagine, Glutamine, Serine, Threonine and Tyrosine.
  • the term "tumor” generally refers to a neoplasm formed by the proliferation of local tissue cells.
  • the tumor can include a solid tumor.
  • the tumor can include a tumor associated with the expression of Siglec15.
  • the term "tumor associated with expression of Siglec15” generally refers to a tumor in which Siglec15 expression is present.
  • the tumor associated with protein expression of Siglec15 may be a Siglec15 positive tumor.
  • the protein expression of Siglec15 on the tumor cell surface or in the tumor microenvironment was approximately 1%, 5%, 10%, 15%, 20%, 25%, 30% higher than normal cells, 35%, 40%, 50%, 60%, 70%, 80% or higher.
  • solid tumor generally refers to a tangible mass that can be detected clinically (eg, X-ray, CT scan, ultrasound, or palpation).
  • the solid tumor can include colon cancer.
  • the term “immunoconjugate” generally refers to a conjugate formed by conjugation (eg, covalently via a linker molecule) of the other therapeutic agent to the isolated antigen-binding protein, the conjugation
  • the other therapeutic agent can be delivered to target cells (eg, tumor cells) by specific binding of the isolated antigen-binding protein to an antigen on the target cell.
  • the antigen may also be secreted by the target cell and located in the space outside the target cell.
  • the term "subject” generally refers to a human or non-human animal, including but not limited to cats, dogs, horses, pigs, cows, sheep, rabbits, mice, rats or monkeys.
  • nucleic acid molecule generally refers to an isolated form of nucleotides, deoxyribonucleotides or ribonucleotides of any length, isolated from their natural environment or artificially synthesized, or analogs thereof.
  • the term "vector” generally refers to a nucleic acid molecule capable of transporting another nucleic acid to which it is linked.
  • the vector can transfer the inserted nucleic acid molecule into and/or between cells.
  • the vectors may include vectors primarily for the insertion of DNA or RNA into cells, vectors primarily for replication of DNA or RNA, and vectors primarily for expression of transcription and/or translation of DNA or RNA.
  • the vector may be a polynucleotide capable of being transcribed and translated into a polypeptide when introduced into a suitable cell.
  • the vector can produce the desired expression product by culturing suitable cells containing the vector.
  • the vector may comprise a lentiviral vector.
  • the term "cell” generally refers to a plasmid or vector that can or has contained a nucleic acid molecule described herein, or an individual cell capable of expressing a polypeptide described herein or an antigen binding protein described herein , cell lines or cell cultures.
  • the cells may include progeny of a single cell. Due to natural, accidental or deliberate mutations, the progeny cells may not necessarily be morphologically or genomically identical to the original parental cells, but are capable of expressing the polypeptides or antigen-binding proteins described herein.
  • the cells can be obtained by transfecting cells in vitro using the vectors described herein.
  • the cells may be prokaryotic cells (eg E.
  • the cells can be immune cells.
  • the immune cells can be selected from the group consisting of T cells, B cells, natural killer cells (NK cells), macrophages, NKT cells, monocytes, dendritic cells, granulocytes, lymphocytes, leukocytes and /or peripheral blood mononuclear cells.
  • treating generally refers to: (i) preventing the occurrence of a disease, disorder or condition in a patient who may be susceptible to a disease, disorder and/or condition but has not been diagnosed with the disease; (ii) inhibiting the disease , disease or condition, i.e. arresting its development; and (iii) alleviating the disease, disorder or condition, i.e. causing the disease, disorder and/or condition and/or symptoms associated with the disease, disorder and/or condition subsided.
  • polypeptide polypeptide
  • peptide protein
  • protein protein
  • proteins are used interchangeably and generally refer to polymers of amino acids of any length.
  • the polymer may be linear or branched, it may contain modified amino acids, and it may be interrupted by non-amino acids. These terms also encompass amino acid polymers that have been modified. These modifications may include: disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation (eg, binding to labeling components).
  • amino acid includes natural and/or non-natural or synthetic amino acids, including glycine and D and L optical isomers, as well as amino acid analogs and peptidomimetics.
  • polynucleotide used interchangeably and generally refer to nucleosides of any length Polymeric forms of acids, such as deoxyribonucleotides or ribonucleotides, or analogs thereof.
  • a polynucleotide can have any three-dimensional structure and can perform any function, known or unknown.
  • polynucleotides coding or non-coding regions of genes or gene fragments, multiple loci (one locus) defined by ligation analysis, exons, introns, messenger RNA (mRNA), Transfer RNA, ribosomal RNA, short interfering RNA (siRNA), short hairpin RNA (shRNA), micro-RNA (miRNA), ribozyme, cDNA, recombinant polynucleotide, branched polynucleotide, plasmid, vector, any sequence of isolated DNA, isolated RNA of any sequence, nucleic acid probes, and primers.
  • mRNA messenger RNA
  • Transfer RNA Transfer RNA
  • ribosomal RNA short interfering RNA
  • shRNA short hairpin RNA
  • miRNA micro-RNA
  • ribozyme ribozyme
  • cDNA recombinant polynucleotide
  • branched polynucleotide plasmid
  • vector any sequence
  • a polynucleotide may contain one or more modified nucleotides, such as methylated nucleotides and nucleotide analogs. If present, modification of the nucleotide structure can be performed before or after polymer assembly. The sequence of nucleotides can be interrupted by non-nucleotide components. Polynucleotides can be further modified after polymerization, such as by conjugation to labeled components.
  • K D (likewise, “K D " or “K D ”) generally refers to "affinity constant” or “equilibrium dissociation constant” and refers to in titration measurements at equilibrium, or Value obtained by dividing the dissociation rate constant (kd) by the association rate constant (ka).
  • Association rate constants (ka), dissociation rate constants (kd), and equilibrium dissociation constants (KD) are used to represent binding of a binding protein (eg, the isolated antigen-binding proteins described herein) to antigen (eg, Siglec15 protein) Affinity. Methods for determining association and dissociation rate constants are well known in the art.
  • the KD value can be determined by Biacore (Biomolecular Interaction Analysis) (eg, an instrument available from BIAcore International AB, aGE Healthcare company, Uppsala, Sweden), or can be detected using other experimental approaches and instruments such as Octet.
  • the KD value can also be determined using KinExA (Kinetic Exclusion Assay) available from Sapidyne Instruments (Boise, Idaho), or using a surface plasmon resonance (SPR) instrument.
  • the K D value can also be determined by an amine coupling kit.
  • the term “comprising” generally means including the expressly specified features, but not excluding other elements. In certain instances, “comprising” also encompasses including only the specified components. For example, to include is also expressed as also to mean “consisting of.”
  • the application provides an isolated antigen-binding protein that can have a KD value of about 4E- 09M or less in a Biacore assay (eg, the KD is not higher than about 4E- 09M , not higher than About 3.5E-09M, not higher than about 3E-09M, not higher than about 2.5E-09M, not higher than about 2E-09M, not higher than about 1.5E-09M, not higher than about 1E-09M, not higher at about 9E-10M, not higher than about 5E-10M, not higher than about 1E-10M, not higher than about 5E-11M, not higher than about 1E-11M or not higher than 5E-12M or less) with human Siglec15 protein specific binding.
  • a Biacore assay eg, the KD is not higher than about 4E- 09M , not higher than About 3.5E-09M, not higher than about 3E-09M, not higher than about 2.5E-09M, not higher than about 2E-09M, not higher than about
  • the isolated antigen-binding protein can have an EC 50 value of about 0.1 ⁇ g/ml or less (eg, the EC50 value is not higher than about 0.1 ⁇ g/ml, not higher than About 99ng/ml, no more than about 95ng/ml, no more than about 90ng/ml, no more than about 85ng/ml, no more than about 80ng/ml, no more than about 75ng/ml, no more than about 70ng /ml, not higher than about 65 ng/ml, not higher than about 60 ng/ml, not higher than about 55 ng/ml, or not higher than about 50 ng/ml or less) specifically binds to human Siglec15 expressed on CHOK1 cells.
  • the EC50 value is not higher than about 0.1 ⁇ g/ml, not higher than About 99ng/ml, no more than about 95ng/ml, no more than about 90ng/ml, no more than about 85ng/ml, no more than about 80ng/m
  • the isolated antigen-binding protein can have an EC 50 value of about 0.1 ⁇ g/ml or less in an ELISA assay (eg, the EC 50 value is not higher than about 0.1 ⁇ g/ml, not higher than About 99ng/ml, no more than about 95ng/ml, no more than about 90ng/ml, no more than about 85ng/ml, no more than about 80ng/ml, no more than about 75ng/ml, no more than about 70ng /ml, no more than about 65 ng/ml, no more than about 60 ng/ml, no more than about 55 ng/ml, or no more than about 50 ng/ml or less) specifically binds to monkey Siglec15.
  • the EC 50 value is not higher than about 0.1 ⁇ g/ml, not higher than About 99ng/ml, no more than about 95ng/ml, no more than about 90ng/ml, no more than about 85ng/ml, no more than about 80ng/
  • the isolated antigen binding protein can relieve the proliferation inhibition of PBMC in blood by Siglec15.
  • the isolated antigen binding protein can promote the proliferation of CD8 and CD4 T cells.
  • the isolated antigen binding protein can promote the expression of IFN- ⁇ .
  • the application provides an isolated antigen binding protein, which can comprise at least one CDR in the VH of the variable region of the antibody heavy chain, and the VH can comprise the amino acid sequence shown in SEQ ID NO:72.
  • the VH may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 29-32.
  • the HCDR of the isolated antigen-binding protein can be divided in any form, as long as the VH is the same as the amino acid sequence shown in any one of SEQ ID NOs: 29-32, the HCDR obtained by dividing in any form can be fall within the scope of protection of this application.
  • the CDRs of antibodies are part of the variable region. Amino acid residues in this region can make contact with the antigen or antigenic epitope.
  • Antibody CDRs can be determined by a variety of coding systems, such as CCG, Kabat, Chothia, IMGT, AbM, Kabat/Chothia, etc. in combination. These coding systems are known in the art, see eg http://www.bioinf.org.uk/abs/index.html#kabatnum. Those skilled in the art can use different coding systems to determine the CDR regions according to the sequence and structure of the antibody. Using different coding systems, there may be differences in the CDR regions.
  • the CDRs encompass CDR sequences that are divided according to any CDR division; variants thereof are also encompassed, the variants comprising substitution, deletion and/or addition of one or more amino acids to the amino acid sequence of the CDR .
  • variants thereof are also encompassed, the variants comprising substitution, deletion and/or addition of one or more amino acids to the amino acid sequence of the CDR .
  • homologues thereof are also encompassed, which may be at least about 85% (e.g., at least about 85%, about 90%, about 91%, about 92%, about 92%, amino acid sequences of about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence homology.
  • the isolated antigen binding proteins described herein are defined by the Kabat coding system.
  • the antigen binding protein may comprise a heavy chain variable region VH, and the VH may comprise at least one, two or three of HCDR1, HCDR2 and HCDR3.
  • the HCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 13.
  • the HCDR3 sequence of the antigen binding protein can be defined according to the Kabat coding system.
  • the HCDR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:9.
  • the HCDR2 sequence of the antigen binding protein can be defined according to the Kabat coding system.
  • the HCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:4.
  • the HCDRl sequence of the antigen binding protein can be defined according to the Kabat coding system.
  • the HCDR1 of the antigen binding protein can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; and the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13 amino acid sequence shown.
  • the antigen binding protein can include antibodies AbO to Ab9 or antigen binding fragments having the same HCDR3 therewith (eg, having the same HCDR1-3).
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:64.
  • H-FR1 of the antigen binding protein has an amino acid substitution (eg, conservative amino acid substitution, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 64: X 2 , X 9 , X 16 , X 17 and X 20 .
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 1-3.
  • the H-FR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:65.
  • H-FR2 of the antigen-binding protein has an amino acid substitution (eg, conservative amino acid substitution, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 65: X 3 , X 8 and X 11 .
  • WV X 3 QAPG X 8 GL X 11 WMG (SEQ ID NO: 65), wherein X 3 can be K or R, X 8 can be K or Q, and X 11 can be E or K.
  • the H-FR2 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 5-8.
  • the H-FR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:66.
  • H-FR3 of the antigen binding protein has amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 66: X 3 , X 7 , X 10 , X 12 , X 18 , X 19 , X 22 , X 27 and X 29 .
  • RF X 3 FSL X 7 TS X 10 S X 12 AYLQI X 18 X 19 LK X 22 EDTA X 27 Y X 29 CAR (SEQ ID NO: 66), wherein X 3 can be A or V and X 7 can be D or E , X 10 can be A or V, X 12 can be M or T, X 18 can be N or S, X 19 can be N or S, X 22 can be A or N, X 27 can be T or V and X 29 can be F or Y.
  • the H-FR3 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 10-12.
  • the H-FR4 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:67.
  • H-FR4 of the antigen binding protein has an amino acid substitution (eg, conservative amino acid substitution, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 67: X 6 and X 7 .
  • WGQGT X 6 X 7 TVSS (SEQ ID NO: 67), wherein X 6 can be L or T and X 7 can be L or V.
  • the H-FR4 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 14 and 15.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 64; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 65; the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO:66; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:67.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 1-3; the H-FR2 may comprise any one of SEQ ID NOs: 5-8 The amino acid sequence shown in item; the H-FR3 may comprise the amino acid sequence shown in any one of SEQ ID NOs: 10-12; and the H-FR4 may comprise any one of SEQ ID NOs: 14 and 15 amino acid sequence shown.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 1; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 5; the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 10; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 14.
  • the antigen-binding protein may comprise antibody Ab0 or an antigen-binding fragment thereof having the same H-FR1-4.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 2; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 6; the H-FR3 can comprise the amino acid sequence shown in SEQ ID NO: 11; and the H-FR4 can comprise the amino acid sequence shown in SEQ ID NO: 15.
  • the antigen-binding protein may include antibodies Ab1, Ab2, Ab3, or antigen-binding fragments thereof having the same H-FR1-4.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 2; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 7; the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 12; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 15.
  • the antigen binding protein may comprise antibodies Ab4, Ab5, Ab6 or antigen binding fragments thereof having the same H-FR1-4.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 3; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 8; the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 12; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 15.
  • the antigen binding protein may comprise antibodies Ab7, Ab8, Ab9 or antigen binding fragments thereof having the same H-FR1-4.
  • the antigen binding protein may comprise a heavy chain variable region, and the heavy chain variable region may comprise the amino acid sequence shown in SEQ ID NO:72.
  • the antigen-binding protein comprises a VH having amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group consisting of: X2 , X9 , X16 , X17 , X20 , X38 , X43 , X46 , X69 , X73 , X76 , X78 , X84 , X85 , X88 , X93 , X95 , X 115 and X 116 .
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 29-32.
  • the antigen binding protein may comprise a heavy chain constant region, which may include an IgG-derived constant region or an IgY-derived constant region.
  • the heavy chain constant region may include a constant region derived from IgGl, IgG2, IgG3 or IgG4.
  • the heavy chain constant region of the antigen binding protein may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 39-42.
  • the antigen binding protein may comprise at least one CDR in the variable region VL of the antibody light chain, and the VL may comprise the amino acid sequence shown in SEQ ID NO:73.
  • the VL may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 33-36.
  • the LCDR of the isolated antigen-binding protein can be divided in any form, as long as the VL is the same as the amino acid sequence shown in any one of SEQ ID NOs: 33-36, the LCDR obtained by dividing in any form can be fall within the scope of protection of this application.
  • the antigen binding protein may comprise a light chain variable region VL, and the VL may comprise at least one, at least two or at least three of LCDR1, LCDR2 and LCDR3.
  • the LCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the LCDR3 of the antigen binding protein can be defined according to the Kabat numbering system.
  • the LCDR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 22.
  • the LCDR2 of the antigen binding protein can be defined according to the Kabat numbering system.
  • the LCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 19.
  • the LCDR1 of the antigen binding protein can be defined according to the Kabat numbering system.
  • the antigen binding protein can include antibodies Ab0 to Ab9 or antigen binding fragments that have the same LCDR3 (eg, have the same LCDR1-3).
  • the VL of the antigen binding protein may comprise the framework regions L-FR1, L-FR2, L-FR3 and L-FR4.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:68.
  • L-FR1 of the antigen binding protein has amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 68: X 5 , X 15 , X 18 and X 19 .
  • DIQM X 5 QSPSSLSAS X 15 GD X 18 X 19 TITC (SEQ ID NO: 68), wherein X 5 can be N or T, X 15 can be L or V, X 18 can be R or T and X 19 can be I or V.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 16-18.
  • the L-FR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:69.
  • the L-FR2 of the antigen-binding protein has amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 69: X 8 and X 9 .
  • WYQQKPG X 8 X 9 PKLLIY (SEQ ID NO: 69), wherein X 8 can be K or N and X 9 can be A or I.
  • the L-FR2 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 20 and 21.
  • the L-FR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:70.
  • L-FR3 of the antigen binding protein has amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 70: X 12 , X 14 and X 27 .
  • the L-FR3 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 23-25.
  • the L-FR4 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:71.
  • L-FR4 of the antigen binding protein has an amino acid substitution (eg, conservative amino acid substitution, etc.) at one or more amino acids selected from the group compared to the sequence shown in SEQ ID NO: 71: X 7 .
  • FGGGTK X7EIK (SEQ ID NO: 71 ), wherein X7 can be L or V.
  • the L-FR4 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 27 and 28.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 68; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 69; the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 70; and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 71.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 16-18; the L-FR2 may comprise any one of SEQ ID NOs: 20 and 21 The amino acid sequence shown in item; the L-FR3 may comprise the amino acid sequence shown in any one of SEQ ID NOs: 23-25; and the L-FR4 may comprise any one of SEQ ID NOs: 27-28 amino acid sequence shown.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 16; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 20; the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 23; and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 27.
  • the antigen binding protein may comprise antibody Ab0 or an antibody having the same L-FR1-4 therewith.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 17; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 21; the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 24; and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 28.
  • the antigen binding protein may include antibodies Ab1, Ab4, Ab7 or antibodies having the same H-FR1-4 therewith.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 18; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 21; the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 25; and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 28.
  • the antigen binding protein may include antibodies Ab2, Ab5, Ab8 or antibodies having the same L-FR1-4 therewith.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 18; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 20; the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 25; and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 28.
  • the antigen binding protein may comprise antibodies Ab3, Ab6, Ab9 or antibodies having the same H-FR1-4 therewith.
  • the antigen binding protein may comprise a light chain variable region VL, and the VL may comprise the amino acid sequence shown in SEQ ID NO:73.
  • the VL of the antigen binding protein has an amino acid substitution (eg, conservative amino acid substitution, etc.) at one or more amino acids selected from the group consisting of: X 5 , X compared to the sequence shown in SEQ ID NO: 73 15 , X 18 , X 19 , X 42 , X 43 , X 68 , X 70 , X 83 , X 104 .
  • X 5 can be N or T
  • X 15 can be L or V
  • X 18 can be R or T
  • X 19 can be I or V
  • X 42 can be K or N
  • X 43 can be A or I
  • X 68 can be G or R
  • X 70 can be D or G
  • X 83 can be F or I
  • X 104 can be L or V.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 33-36.
  • the antigen binding protein may comprise a light chain constant region, which may include an Ig ⁇ -derived constant region or an Ig ⁇ -derived constant region.
  • the light chain constant region can include a constant region derived from IgK.
  • the light chain constant region of the antigen binding protein comprises the amino acid sequence set forth in any one of SEQ ID NO:43.
  • the antigen binding protein may comprise HCDR1-3 and LCDR1-3.
  • the HCDR1 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:4; the HCDR2 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:9; the HCDR3 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:9; comprise the amino acid sequence shown in SEQ ID NO: 13; the LCDR1 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 of the antigen-binding protein may comprise the amino acid shown in SEQ ID NO: 22 sequence; LCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:26.
  • the antigen-binding protein can include antibodies Ab0-Ab9 or antigen-binding fragments thereof having the same HCDR3 (e.g., having the same HCDR1-3) and LCDR3 (e.g., having the same LCDR1-3).
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the antigen binding protein may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region of the antigen binding protein may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:1; the H-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:5; the H-FR3 may comprise the amino acid sequence set forth in SEQ ID NO:10
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:29.
  • the antigen binding protein may comprise antibody AbO or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:33.
  • the antigen binding protein may comprise antibody AbO or an antigen binding protein having the same light chain variable region.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:2; the H-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:6; the H-FR3 may comprise the amino acid sequence set forth in SEQ ID NO:11
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:30.
  • the antigen binding protein may include antibody Ab1 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:34.
  • the antigen binding protein may comprise antibody Ab1 or an antigen binding protein having the same light chain variable region therewith.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:2; the H-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:6; the H-FR3 may comprise the amino acid sequence set forth in SEQ ID NO:11
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:30.
  • the antigen binding protein may include antibody Ab2 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:35.
  • the antigen binding protein may include antibody Ab2 or an antigen binding protein having the same light chain variable region therewith.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:2; the H-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:6; the H-FR3 may comprise the amino acid sequence set forth in SEQ ID NO:11
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:30.
  • the antigen binding protein may comprise the antigen binding fragment Ab3 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:36.
  • the antigen binding protein may comprise antibody Ab3 or an antigen binding protein having the same light chain variable region.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 can comprise the amino acid sequence set forth in SEQ ID NO:2; the H-FR2 can comprise the amino acid sequence set forth in SEQ ID NO:7; the H-FR3 can comprise the amino acid sequence set forth in SEQ ID NO:12
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:31.
  • the antigen binding protein may include antibody Ab4 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:34.
  • the antigen binding protein may include antibody Ab4 or an antigen binding protein having the same light chain variable region.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the antigen binding protein may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region of the antigen binding protein may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 can comprise the amino acid sequence set forth in SEQ ID NO:2; the H-FR2 can comprise the amino acid sequence set forth in SEQ ID NO:7; the H-FR3 can comprise the amino acid sequence set forth in SEQ ID NO:12
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:31.
  • the antigen binding protein may comprise antibody Ab5 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:35.
  • the antigen binding protein may comprise antibody Ab5 or an antigen binding protein having the same light chain variable region therewith.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 can comprise the amino acid sequence set forth in SEQ ID NO:2; the H-FR2 can comprise the amino acid sequence set forth in SEQ ID NO:7; the H-FR3 can comprise the amino acid sequence set forth in SEQ ID NO:12
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:31.
  • the antigen binding protein may comprise antibody Ab6 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:36.
  • the antigen binding protein may comprise antibody Ab6 or an antigen binding protein having the same light chain variable region therewith.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 can comprise the amino acid sequence set forth in SEQ ID NO:3; the H-FR2 can comprise the amino acid sequence set forth in SEQ ID NO:8; the H-FR3 can comprise the amino acid sequence set forth in SEQ ID NO:12
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:32.
  • the antigen binding protein may comprise antibody Ab7 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:34.
  • the antigen binding protein may comprise antibody Ab7 or an antigen binding protein having the same light chain variable region.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 can comprise the amino acid sequence set forth in SEQ ID NO:3; the H-FR2 can comprise the amino acid sequence set forth in SEQ ID NO:8; the H-FR3 can comprise the amino acid sequence set forth in SEQ ID NO:12
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:32.
  • the antigen binding protein may comprise antibody Ab8 or an antigen binding protein having the same heavy chain variable region therewith.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:35.
  • the antigen binding protein may comprise antibody Ab8 or an antigen binding protein having the same light chain variable region therewith.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the H-FR1 can comprise the amino acid sequence set forth in SEQ ID NO:3; the H-FR2 can comprise the amino acid sequence set forth in SEQ ID NO:8; the H-FR3 can comprise the amino acid sequence set forth in SEQ ID NO:12
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:32.
  • the antigen binding protein may comprise antibody Ab9 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence set forth in SEQ ID NO:36.
  • the antigen binding protein can include antibody Ab9 or an antigen binding protein having the same light chain variable region.
  • the isolated antigen binding protein may also compete for binding to the human Siglec15 protein with a reference antibody, which may comprise a heavy chain variable region VH, which may comprise HCDR1, HCDR2 and HCDR3 at least one, two, or three of them.
  • a reference antibody which may comprise a heavy chain variable region VH, which may comprise HCDR1, HCDR2 and HCDR3 at least one, two, or three of them.
  • the HCDR3 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 13.
  • the sequence of the HCDR3 of the reference antibody can be defined according to the Kabat coding system.
  • the HCDR2 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO:9.
  • the sequence of the HCDR2 of the reference antibody can be defined according to the Kabat coding system.
  • the HCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO:4.
  • the sequence of the HCDR1 of the reference antibody can be defined according to the Kabat coding system.
  • HCDR1 of the reference antibody can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; and the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13 amino acid sequence shown.
  • the reference antibody can include antibodies Ab0 to Ab9 or antigen binding proteins that have the same HCDR3 (eg, have the same HCDR1-3).
  • the reference antibody may comprise a heavy chain variable region, and the heavy chain variable region may comprise the amino acid sequence shown in SEQ ID NO:72.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence shown in any one of SEQ ID NOs: 29-32.
  • the reference antibody may comprise a heavy chain constant region, which may include an IgG-derived constant region or an IgY-derived constant region.
  • the heavy chain constant region of the reference antibody may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 39-42.
  • the reference antibody may comprise a light chain variable region VL, which may comprise LCDR1, LCDR2 and LCDR3.
  • the LCDR3 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO:26.
  • the sequence of LCDR3 of the reference antibody can be defined according to the Kabat coding system.
  • the LCDR2 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 22.
  • the sequence of LCDR2 of the reference antibody can be defined according to the Kabat coding system.
  • the LCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 19.
  • the sequence of LCDR1 of the reference antibody can be defined according to the Kabat coding system.
  • the reference antibody can include antibodies AbO to Ab9 or antigen binding proteins that have the same LCDR3 (eg, have the same LCDR1-3).
  • the reference antibody may comprise a light chain variable region, and the light chain variable region may comprise the amino acid sequence shown in SEQ ID NO:73.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence shown in any one of SEQ ID NOs: 33-36.
  • the reference antibody may comprise a light chain constant region comprising a constant region derived from Ig ⁇ or a constant region derived from Ig ⁇ .
  • the light chain constant region of the reference antibody comprises the amino acid sequence set forth in any one of SEQ ID NO:43.
  • the reference antibody may comprise HCDR1-3 and LCDR1-3.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:4; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:9; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:13;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 19; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 22; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 26.
  • the reference antibody can include antibodies Ab0-Ab9 or antigen binding proteins that have the same HCDR3 (eg, have the same HCDR1-3) and LCDR3 (eg, have the same LCDR1-3).
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:29.
  • the reference antibody may comprise antibody AbO or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:33.
  • the reference antibody may comprise antibody AbO or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab0 or an antigen binding protein having the same heavy chain variable region and light chain variable region.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:30.
  • the reference antibody may comprise antibody Ab1 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:34.
  • the reference antibody may comprise antibody Ab1 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab1 or an antigen binding protein having the same heavy chain variable region and light chain variable region.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:30.
  • the reference antibody may comprise antibody Ab2 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:35.
  • the reference antibody may comprise antibody Ab2 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab2 or an antigen binding protein having the same heavy chain variable region and light chain variable region.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:30.
  • the reference antibody may comprise antibody Ab3 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:36.
  • the reference antibody may comprise antibody Ab3 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab3 or an antigen binding protein having the same heavy chain variable region and light chain variable region.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:31.
  • the reference antibody may comprise antibody Ab4 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:34.
  • the reference antibody may comprise antibody Ab4 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab4 or an antigen binding protein having the same heavy and light chain variable regions.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:31.
  • the reference antibody may comprise antibody Ab5 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:35.
  • the reference antibody may comprise antibody Ab5 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab5 or an antigen binding protein having the same heavy chain variable region and light chain variable region.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:31.
  • the reference antibody may comprise antibody Ab6 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:36.
  • the reference antibody may comprise antibody Ab6 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab6 or an antigen binding protein having the same heavy and light chain variable regions.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:32.
  • the reference antibody may comprise an antigen binding protein with which antibody Ab7 has the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:34.
  • the reference antibody may comprise antibody Ab7 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may include antibody Ab7 or an antigen binding protein having the same heavy and light chain variable regions.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:32.
  • the reference antibody may comprise antibody Ab8 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:35.
  • the reference antibody may comprise antibody Ab8 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may include antibody Ab8 or an antigen binding protein having the same heavy and light chain variable regions.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:32.
  • the reference antibody may comprise antibody Ab9 or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:36.
  • the reference antibody may comprise antibody Ab9 or an antigen binding protein having the same light chain variable region.
  • the reference antibody may comprise antibody Ab9 or an antigen binding protein having the same heavy and light chain variable regions.
  • the application provides one or more polypeptides that may comprise the isolated antigen binding proteins of the application.
  • the polypeptide may comprise a fusion protein.
  • the polypeptides can include multispecific antibodies (eg, bispecific antibodies).
  • the application provides one or more immunoconjugates that can comprise the isolated antigen binding proteins of the application.
  • the immunoconjugate may further comprise a pharmaceutically acceptable therapeutic agent, marker and/or detection agent.
  • the present application also provides isolated one or more nucleic acid molecules that encode the isolated antigen-binding proteins described herein.
  • each of the one or more nucleic acid molecules may encode the entire antigen binding protein, or may encode a portion thereof (eg, HCDR1-3, one of the heavy chain variable regions, or variety).
  • the products encoded by the nucleic acid molecules together can form a functional (eg, can bind Siglec15) isolated antigen-binding protein of the present application.
  • the nucleic acid molecules described herein can be isolated. For example, it may be produced or synthesized by: (i) amplified in vitro, for example by polymerase chain reaction (PCR) amplification, (ii) recombinantly produced by cloning, (iii) purified either (iv) synthetic, eg by chemical synthesis.
  • the isolated nucleic acid can be a nucleic acid molecule prepared by recombinant DNA techniques.
  • nucleic acids encoding the isolated antigen-binding proteins can be prepared by various methods known in the art, including but not limited to, using reverse transcription PCR and PCR to obtain the isolated antigen-binding proteins described herein protein nucleic acid molecules.
  • the application provides one or more vectors comprising one or more nucleic acid molecules described herein.
  • One or more of the nucleic acid molecules may be included in each vector.
  • 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 also contain expression control elements that allow the correct expression of the coding region in an appropriate host.
  • control elements are well known to those of skill in the art, and may include, for example, promoters, ribosome binding sites, enhancers, and other control elements that regulate gene transcription or mRNA translation, and the like.
  • the expression control sequence is a tunable element.
  • the specific structure of the expression control sequence may vary depending on species or cell type function, but typically comprises 5' untranslated and 5' and 3' untranslated sequences involved in transcription and translation initiation, respectively, such as the TATA box, plus Cap sequences, CAAT sequences, etc.
  • a 5' non-transcribed expression control sequence may comprise a promoter region, which may comprise a promoter sequence for transcriptional control of a functionally linked nucleic acid.
  • the expression control sequences may also include enhancer sequences or upstream activator sequences.
  • suitable promoters may include, for example, the promoters for SP6, T3 and T7 polymerases, the human U6 RNA promoter, the CMV promoter, and artificial hybrid promoters thereof (such as CMV), wherein the promoter's A portion may be fused to a portion of the gene promoter for other cellular proteins (eg, human GAPDH, glyceraldehyde-3-phosphate dehydrogenase), which may or may not contain additional introns.
  • One or more nucleic acid molecules described herein can be operably linked to the expression control element.
  • Lentiviral vectors are retroviral vectors capable of transducing or infecting non-dividing cells and typically producing higher viral titers. Lentiviral vectors may comprise long terminal repeats 5'LTR and truncated 3'LTRs, RREs, rev response elements (cPPT), central termination sequences (CTS) and/or post-translational regulatory elements (WPRE). The vectors described herein can be introduced into cells.
  • the application provides a cell.
  • the cells may comprise the isolated antigen binding proteins described herein, the polypeptides, the immunoconjugates, one or more nucleic acid molecules and/or one or more vectors described herein .
  • each or each cell may contain one or one nucleic acid molecule or vector described herein.
  • each or each cell can comprise a plurality (eg, 2 or more) or more (eg, 2 or more) of the nucleic acid molecules or vectors described herein.
  • the cells may include immune cells.
  • the cells can include immune cells.
  • the cells can include T cells, B cells, natural killer (NK) cells, macrophages, NKT cells, monocytes, dendritic cells, granulocytes, lymphocytes, leukocytes, and/or peripheral blood mononuclear cells cell.
  • NK natural killer
  • the application provides a pharmaceutical composition.
  • the pharmaceutical composition may comprise the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell, and/or the isolated antigen-binding protein described herein. or pharmaceutically acceptable adjuvants and/or excipients.
  • the pharmaceutically acceptable adjuvants may include buffers, antioxidants, preservatives, low molecular weight polypeptides, proteins, hydrophilic polymers, amino acids, sugars, chelating agents, counterions, metal complexes and /or nonionic surfactants. Unless incompatible with the cells described herein, any conventional medium or agent is contemplated for use in the pharmaceutical compositions of the present application.
  • the pharmaceutically acceptable excipients may include additives other than the main drug in the pharmaceutical preparation, and may also be referred to as excipients.
  • the excipients may include binders, fillers, disintegrants, lubricants in the tablet.
  • the excipients may include wine, vinegar, medicinal juice, etc. in Chinese medicine pills.
  • the excipient may comprise a base part of a semisolid formulation ointment, cream.
  • the excipients may include preservatives, antioxidants, flavors, fragrances, solubilizers, emulsifiers, solubilizers, osmo-regulators, colorants in liquid formulations.
  • the application provides a pharmaceutical combination comprising the isolated antigen binding protein and an immune checkpoint inhibitor.
  • the immune checkpoint inhibitor may include substances that inhibit the interaction of PD-1/PD-L1.
  • the immune checkpoint inhibitor can be selected from the group consisting of PD-1/PD-L1 blockers, PD-1 antagonists, PD-L1 antagonists, PD-1 inhibitors and PD-L1 inhibitors.
  • the PD-1/PD-L1 blocker may be selected from the group consisting of BMS202 (PD-1/PD-L1 inhibitor 2), BMS-1 (PD-1/PD-L1 inhibitor 1), PD-1/PD-L1 inhibitor 3, BMS-1166 and BMS-1001.
  • the PD-1 inhibitor can include an anti-PD-1 antibody.
  • the PD-L1 inhibitor can include an anti-PD-L1 antibody.
  • the anti-PD-1 antibody may be selected from the group consisting of Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalizumab) ), Sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-L1 antibody may be selected from the group consisting of Durvalumab (druvalumab), Atezolizumab (atezolizumab), and avelumab (avelumab).
  • the anti-PD-1 antibody may comprise the HCDR3 of an antibody selected from the group consisting of Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-1 antibody may comprise the HCDR2 of an antibody selected from the group consisting of: Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-1 antibody may comprise HCDR1 of an antibody selected from the group consisting of Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-1 antibody may comprise LCDR3 of an antibody selected from the group consisting of: Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-1 antibody may comprise LCDR2 of an antibody selected from the group consisting of: Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-1 antibody may comprise LCDR1 of an antibody selected from the group consisting of: Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-1 antibody may comprise the VH of an antibody selected from the group consisting of Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-1 antibody may comprise the VL of an antibody selected from the group consisting of: Nivolumab (nivolumab), Pembrolizumab (pembrolizumab), Camrelizumab (camrelizumab), Toripalimab (toripalimab), sintilimab (sintilimab) and Tislelizumab (tislelizumab).
  • the anti-PD-L1 antibody may comprise the HCDR3 of an antibody selected from the group consisting of Durvalumab (durvalumab), Atezolizumab (atezolizumab), and avelumab (avelumab).
  • the anti-PD-L1 antibody may comprise the HCDR2 of an antibody selected from the group consisting of Durvalumab (durvalumab), Atezolizumab (atezolizumab), and avelumab (avelumab).
  • the anti-PD-L1 antibody may comprise HCDR1 of an antibody selected from the group consisting of Durvalumab (druvalumab), Atezolizumab (atezolizumab), and avelumab (avelumab).
  • the anti-PD-L1 antibody may comprise LCDR3 of an antibody selected from the group consisting of Durvalumab (durvalumab), Atezolizumab (atezolizumab), and avelumab (avelumab).
  • the anti-PD-L1 antibody may comprise LCDR2 of an antibody selected from the group consisting of Durvalumab (durvalumab), Atezolizumab (atezolizumab), and avelumab (avelumab).
  • the anti-PD-L1 antibody may comprise the VH of an antibody selected from the group consisting of Durvalumab (durvalumab), Atezolizumab (atezolizumab) and avelumab (avelumab).
  • the anti-PD-L1 antibody may comprise the VL of an antibody selected from the group consisting of Durvalumab (durvalumab), Atezolizumab (atezolizumab), and avelumab (avelumab).
  • the anti-PD-1 antibody may comprise HCDR3, and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:46.
  • the anti-PD-1 antibody comprises HCDR2, and the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:45.
  • the anti-PD-1 antibody comprises HCDR1, and the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:44.
  • the anti-PD-1 antibody may comprise a heavy chain variable region VH, and the VH may comprise the amino acid sequence shown in SEQ ID NO:50.
  • the anti-PD-1 antibody may comprise LCDR3, and the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO:49.
  • the anti-PD-1 antibody may comprise LCDR2, and the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO:48.
  • the anti-PD-1 antibody may comprise LCDR1, and the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO:47.
  • the anti-PD-1 antibody may comprise a light chain variable region VL, the VL may comprise LCDR1, LCDR2 and LCDR3, and the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 49; the LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:48; and the LCDR1 may comprise the amino acid sequence set forth in SEQ ID NO:47.
  • the anti-PD-1 antibody can include pembrolizumab or an antibody that has the same LCDR3 (eg, has the same LCDR1-3).
  • the anti-PD-1 antibody may comprise a heavy chain and a light chain, the heavy chain may comprise HCDR1-3 and H-FR1-4, and the light chain may comprise LCDR1-3 and L-FR1 -4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:44; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:45; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:46;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO:47; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO:48; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO:49.
  • the anti-PD-1 antibody can include pembrolizumab or an antigen binding protein having the same HCDR3 (eg, having the same HCDR1-3) and LCDR3 (eg, having the same LCDR1-3).
  • the anti-PD-L1 antibody may comprise HCDR3, and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:60.
  • the anti-PD-L1 antibody comprises HCDR2, and the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:59.
  • the anti-PD-L1 antibody comprises HCDR1, and the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:58.
  • the anti-PD-L1 antibody comprises a heavy chain variable region VH
  • the VH comprises HCDR1, HCDR2 and HCDR3
  • the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 60
  • the HCDR2 comprises SEQ ID NO: 60
  • the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:58.
  • the anti-PD-L1 antibody can include atezolizumab or an antibody having the same HCDR3 (eg, having the same HCDR1-3).
  • the anti-PD-L1 antibody may comprise a heavy chain variable region VH, and the VH may comprise the amino acid sequence shown in SEQ ID NO:54.
  • the anti-PD-L1 antibody may comprise LCDR3, and the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO:63.
  • the anti-PD-L1 antibody may comprise LCDR2, and the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO:62.
  • the anti-PD-L1 antibody may comprise LCDR1, and the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO:61.
  • the anti-PD-L1 antibody may comprise a light chain variable region VL, the VL may comprise LCDR1, LCDR2 and LCDR3, and the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 63; the LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:62; and the LCDR1 may comprise the amino acid sequence set forth in SEQ ID NO:61.
  • the anti-PD-L1 antibody can include atezolizumab or an antibody having the same LCDR3 (eg, having the same LCDR1-3).
  • the anti-PD-L1 antibody may comprise a light chain variable region VL, and the VL may comprise the amino acid sequence shown in SEQ ID NO:55.
  • the anti-PD-L1 antibody may comprise a heavy chain and a light chain, the heavy chain may comprise HCDR1-3 and H-FR1-4, and the light chain may comprise LCDR1-3 and L-FR1 -4.
  • the HCDR1 can comprise the amino acid sequence set forth in SEQ ID NO:58; the HCDR2 can comprise the amino acid sequence set forth in SEQ ID NO:59; the HCDR3 can comprise the amino acid sequence set forth in SEQ ID NO:60;
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO:61; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO:62; the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO:63.
  • the anti-PD-L1 antibody can include atezolizumab or an antigen binding protein having the same HCDR3 (eg, having the same HCDR1-3) and LCDR3 (eg, having the same LCDR1-3).
  • the heavy chain variable region of the anti-PD-L1 antibody may comprise the amino acid sequence shown in SEQ ID NO:54.
  • the anti-PD-L1 antibody may comprise atezolizumab or an antigen binding protein having the same heavy chain variable region.
  • the light chain variable region of the anti-PD-L1 antibody may comprise the amino acid sequence shown in SEQ ID NO:55.
  • the anti-PD-L1 antibody may include atezolizumab or an antigen binding protein having the same light chain variable region.
  • the heavy chain of the anti-PD-L1 antibody may comprise the amino acid sequence set forth in SEQ ID NO:56.
  • the anti-PD-L1 antibody may include atezolizumab or an antigen binding protein having the same heavy chain.
  • the light chain of the anti-PD-L1 antibody may comprise the amino acid sequence shown in SEQ ID NO:57.
  • the anti-PD-L1 antibody may include atezolizumab or an antigen binding protein having the same light chain.
  • the methods may include in vitro methods, ex vivo methods, methods not for diagnostic or therapeutic purposes.
  • the method may include a method for detecting the presence and/or amount of Siglec15 for non-diagnostic purposes, which may include the steps of:
  • the present application provides a kit for Siglec15, which can include the use of the isolated antigen-binding protein or the polypeptide.
  • the kit may further comprise instructions for use describing the method for detecting the presence and/or content of Siglec15.
  • the methods may include in vitro methods, ex vivo methods, methods not for diagnostic or therapeutic purposes.
  • the present application provides the use of the isolated antigen-binding protein or the polypeptide in the preparation of a kit, which can be used for a method for detecting the presence and/or content of Siglec15.
  • the methods may include in vitro methods, ex vivo methods, methods not for diagnostic or therapeutic purposes.
  • the application provides a method of modulating an immune response, comprising administering to a subject in need thereof an effective amount of the isolated antigen binding protein, the polypeptide, the immunoconjugate, The isolated nucleic acid molecule, the carrier, the cell and/or the pharmaceutical composition, and/or the pharmaceutically acceptable therapeutic agent.
  • the method of modulating an immune response may include in vitro methods, ex vivo methods, methods not for diagnostic or therapeutic purposes.
  • the method of modulating an immune response may include in vitro methods, ex vivo methods, methods not for diagnostic or therapeutic purposes.
  • the disease or disorder may include abnormal bone metabolism.
  • the abnormal bone metabolism can include osteoporosis, bone destruction associated with rheumatoid arthritis, cancerous hypercalcemia, bone destruction associated with multiple myeloma or bone metastases from cancer, osteopenia, dental Tooth loss due to periarthritis, osteolysis around artificial joints, bone destruction in chronic osteomyelitis, Paget's disease of bone, renal osteodystrophy, and osteogenesis imperfecta.
  • the abnormal bone metabolism can include osteoporosis.
  • the antigen binding proteins of the present application can be administered alone or in combination with at least one other therapeutic agent for bone-related diseases.
  • the antigen binding proteins of the present application can be administered in combination with a therapeutically effective amount of a therapeutic agent for abnormal bone metabolism.
  • Therapeutic agents that can be administered in combination with the antigen binding proteins of the present application include, but are not limited to: bisphosphonates (eg, alendronate, etidronate, ibandronate, icardronate, paclitaxel diphosphate, risedronate and zoledronate), active vitamin D3, calcitonin and its derivatives, hormones such as estradiol, SERM (selective estrogen receptor modulator), ippro Flavonoids, vitamin K2 (menadione), calcium preparations, PTH (parathyroid hormone), NSAIDs (eg, celecoxib and rofecoxib), soluble TNF receptors (eg, , etanercept), anti-TNF- ⁇ antibody or functional fragment of said antibody (eg, infliximab), anti-PTHrP (parathyroid hormone-related protein)
  • the disease or disorder can include a tumor.
  • the tumor can include a solid tumor.
  • the tumor can include a tumor associated with the expression of Siglec15.
  • the term "tumor associated with the expression of Siglec15" generally refers to tumors in which altered expression of Siglec15 leads to disease progression or evasion of immune surveillance.
  • the "tumor associated with the expression of Siglec15" may be a tumor formed by up-regulation of the expression of Siglec15 leading to disease progression or evasion of immune surveillance.
  • the tumor associated with protein expression of Siglec15 may be a Siglec15 positive tumor.
  • the protein expression of Siglec15 on the tumor cell surface or in the tumor microenvironment was approximately 1%, 5%, 10%, 15%, 20%, 25%, 30% higher than normal cells, 35%, 40%, 50%, 60%, 70%, 80% or higher.
  • the tumor can include colon cancer.
  • the present application provides the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell and /or the use of the pharmaceutical composition in the preparation of a medicament for preventing, relieving and/or treating a disease or condition.
  • the present application provides the use of a pharmaceutical combination in the preparation of a medicament for preventing, alleviating and/or treating a disease or condition.
  • the antigen binding proteins of the present application can be administered in combination with a therapeutically effective amount of a therapeutic agent for abnormal bone metabolism.
  • Therapeutic agents that can be administered in combination with the antigen binding proteins of the present application include, but are not limited to: bisphosphonates (eg, alendronate, etidronate, ibandronate, icadronate, paclitaxel diphosphate, risedronate and zoledronate), active vitamin D3, calcitonin and its derivatives, hormones such as estradiol, SERM (selective estrogen receptor modulator), ippro Flavonoids, vitamin K2 (menadione), calcium preparations, PTH (parathyroid hormone), NSAIDs (eg, celecoxib and rofecoxib), soluble TNF receptors (eg, , etanercept), anti-TNF- ⁇ antibody or functional fragment of said antibody (eg, infliximab), anti-PTHrP (parathyroid hormone-related protein) antibody
  • the disease or disorder may include abnormal bone metabolism.
  • the abnormal bone metabolism may include osteoporosis, bone destruction associated with rheumatoid arthritis, cancerous hypercalcemia, bone destruction associated with multiple myeloma or bone metastases from cancer, osteopenia, dental Tooth loss due to periarthritis, osteolysis around artificial joints, bone destruction in chronic osteomyelitis, Paget's disease of bone, renal osteodystrophy, and osteogenesis imperfecta.
  • the abnormal bone metabolism can include osteoporosis.
  • the antigen binding proteins of the present application can be administered alone or in combination with at least one other therapeutic agent for bone-related diseases.
  • the antigen binding proteins of the present application can be administered in combination with a therapeutically effective amount of a therapeutic agent for abnormal bone metabolism.
  • Therapeutic agents that can be administered in combination with the antigen binding proteins of the present application include, but are not limited to: bisphosphonates (eg, alendronate, etidronate, ibandronate, icardronate, paclitaxel diphosphate, risedronate and zoledronate), active vitamin D3, calcitonin and its derivatives, hormones such as estradiol, SERM (selective estrogen receptor modulator), ippro Flavonoids, vitamin K2 (menadione), calcium preparations, PTH (parathyroid hormone), NSAIDs (eg, celecoxib and rofecoxib), soluble TNF receptors (eg, , etanercept), anti-TNF- ⁇ antibody or functional fragment of said antibody (eg, infliximab), anti-PTHrP (parathyroid hormone-related protein)
  • the disease or disorder can include a tumor.
  • the tumor can include a solid tumor.
  • the tumor can include a tumor associated with the expression of Siglec15.
  • the term "tumor associated with the expression of Siglec15" generally refers to tumors in which altered expression of Siglec15 leads to disease progression or evasion of immune surveillance.
  • the "tumor associated with the expression of Siglec15" may be a tumor formed by up-regulation of the expression of Siglec15 leading to disease progression or evasion of immune surveillance.
  • the tumor associated with protein expression of Siglec15 may be a Siglec15 positive tumor.
  • the tumor can include colon cancer.
  • the effective amount in humans can be extrapolated from the effective amount in experimental animals.
  • Freireich et al. describe the correlation of doses (based on milligrams per square meter of body surface) in animals and humans (Freiheim et al., Cancer Chemother. Rep. 50, 219 (1966)).
  • Body surface area can be approximately determined from the patient's height and weight. See, eg, Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y., 537 (1970).
  • Antigen coating prepare a solution of husiglec-15-hFc antigen (human Siglec-15Fc, SinoBiological, Cat. No. 13976-H02H) diluted in concentration gradient with DPBS solution, and coat the 96-well plate overnight at 4°C.
  • Siglec-15 antibody mouse-human chimeric antibody (5G12, human IgG1), Acrobiosystem) was quantitatively diluted with DPBS (containing 1% BSA) solution, and coated for 1 hour at 37°C.
  • 1.6 ELISA detection Discard the reaction solution, wash it three times with PBST solution; add TMB solution and H 2 SO 4 stop solution successively, and after slight shaking turns yellow, detect the OD value at 450 nm.
  • Antigen coating prepare a concentration gradient dilution of husiglec-15-his antigen (human Siglec-15his, Acrobiosystem, product number: SG5-H52H3) solution with DPBS solution, cyno-siglec-15-hFc (cynomolgus monkey Siglec-15his, Acrobiosystem, product number: SG5-C52H6) took a 96-well plate, added 100 ⁇ l to each well, and coated overnight at 4°C.
  • mice type SJL rat (Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.) immune antigen husiglec-15-Fc antigen immunization method
  • the selected biologically active antibodies can optionally be humanized.
  • Humanization of murine monoclonal antibodies is carried out according to methods disclosed in many literatures in the art. Briefly, CDR grafting can be performed using human antibody constant domains in place of parental (murine antibody) constant domains, selecting human germline antibody sequences based on the homology of the murine and human antibodies. Then, based on the three-dimensional structure of the murine antibody, back-mutation of the amino acid residues of VL and VH can be performed to replace the constant region of the murine antibody with the human constant region to obtain the final humanized binding protein.
  • a plasmid was constructed according to the antigen-binding protein sequence of the present application, transiently expressed in Expi293 cells (Thermo Fisher, Cat. No. A14527CN), and the antibody expression plasmid was extracted using a large-scale extraction kit.
  • Solution 1 Dilute 15 ⁇ g of plasmid with 1 ml of culture medium and mix well.
  • Solution 2 Dilute 60 ⁇ l of transfection reagent with 1 ml of culture medium and mix well.
  • Dialysis suck the high-concentration protein into a dialysis bag and place it in a beaker of 1 ⁇ PBS for dialysis.
  • the purity test using high performance liquid chromatography LC-20AT and gel chromatography column is qualified, and the endotoxin test is qualified.
  • Siglec15 antigen-binding protein The binding ability of Siglec15 antigen-binding protein to human Siglec15 expressed on the surface of CHOK1 cells was determined based on a flow cytometry assay. The binding capacity of different Siglec15 antigen-binding proteins was determined by comparing their binding curves to human Siglec15 expressed on the surface of CHOK1 cells.
  • the cells were resuspended with 2% FBS in PBS, and the median fluorescence value (MFI) of the PE channel was measured by flow cytometry.
  • MFI median fluorescence value
  • Siglec15 In the presence of Siglec15, the Siglec15 antigen-binding protein was co-incubated with PBMCs extracted from human blood for a certain period of time. Finally, Siglec15 was determined by detecting the proliferation of CD4 and CD8 T cells in the co-incubated PBMCs and the expression of IFN- ⁇ in the supernatant of the medium. Whether antigen-binding protein can block the ability of Siglec15 to inhibit the proliferation of PBMCs in the blood of subjects.
  • the cells were resuspended with 2% FBS in PBS, and the median fluorescence intensity (MFI) of the PE channel was measured by flow cytometry.
  • MFI median fluorescence intensity
  • the Siglec15 antigen-binding protein can block the ability of Siglec15 to inhibit the proliferation of PBMCs in the blood of subjects.
  • Siglec15 antigen-binding protein blocks the ability of Siglec15 to inhibit the proliferation of PBMCs in the blood of subjects
  • Biacore was used to detect the binding affinity of different Siglec15 antigen-binding proteins to antigen (Siglec15 protein, human, recombinant (ECD, His Tag), source: Acro, Cat. No.: SG5-C5253) protein.
  • Running reagents containing 10 mM N-(2-hydroxyethyl)piperazine-N-2 sulfonic acid (HEPES), 150 mM sodium chloride (NaCl), 3 mM ethylenediaminetetraacetic acid (EDTA), 0.005% Tween-20 (Tween-20), pH adjusted to 7.4; Human IgG(Fc) Capture Kit (Cat. No. BR-1008-39, GE), including: mouse anti-human IgG(Fc) antibody (0.5mg/mL), immobilized Reagents (10mM Sodium Acetate, pH5.0), Regeneration Reagent (3M Magnesium Chloride); Amino Coupling Kit (Cat. No.
  • mice anti-human IgG (Fc) antibody was injected into the experimental channel (FC2) at a flow rate of 10 ⁇ L/min for about 420 s, and the fixed amount was about 9000 to 14000 RU.
  • the chip was blocked with 1 M ethanolamine at 10 ⁇ L/min for 420 s.
  • the reference channel (FC1) performs the same operation as the test channel (FC2).
  • Antigen was diluted to 200 nM with running reagent. After capturing the antibody, the diluted antigens were sequentially combined for 90 seconds and dissociated for 210 seconds at a flow rate of 30 ⁇ l/min, and then injected into the experimental channel and the reference channel, and each antigen-binding protein was set to a concentration of 0. After each antigen-binding protein analysis, the chip was regenerated with 3M magnesium chloride at a flow rate of 20 ⁇ l/min for 30 seconds to wash away ligand as well as undissociated analyte.
  • Mouse-derived MC38 cells were purchased from Heyuan Biotechnology (Shanghai) Co., Ltd. and routinely subcultured according to the instructions; SPH genetically modified MC38 cells to overexpress human Siglec15, and the cells were named MC38/ hSiglec15 cells were used for subsequent in vivo experiments. The cells were collected by centrifugation, resuspended in serum-free medium and adjusted for cell density. On day 0, the cell suspension was subcutaneously inoculated into the right axilla of wild-type C57BL/6N mice subcutaneously to establish the MC38/hSiglec15 tumor-bearing mouse model.
  • PBMC peripheral mononuclear cells
  • Mouse-derived MC38 cells were purchased from Heyuan Biotechnology (Shanghai) Co., Ltd. and routinely subcultured according to the instructions; MC38 cells were genetically modified to overexpress human-derived Siglec15, and the cell was named MC38/human Siglec15 cells for subsequent in vivo experiments. Cells were collected by centrifugation, resuspended in serum-free medium and adjusted for cell density. On day 0, the cell suspension was subcutaneously inoculated into the right axilla of human PD-1/PD-L1 transgenic mice to establish MC38/human Siglec15 cells. tumor mouse model.
  • mice with tumor volume in the range of 120 mm 3 to 230 mm 3 were selected and grouped by tumor volume (6 mice in each group).
  • Single dose PD-1 antibody (Pembrolizumab, source: Taizhou Baiying Biotechnology Co., Ltd., product number: B2014-CHO); on the 7th, 10th, 14th, 17th, 21st, 24 days of administration, negative control antibody (human IgG), positive control antibody (SPH-Sg-PC-1, source: Hangzhou Haoyang Biotechnology Co., Ltd., product number HSP067-41), antigen-binding protein of the application; during the administration period The changes of tumor volume and body weight of mice in each group were monitored, and the monitoring frequency was 2 times/week for 3 consecutive weeks.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Biochemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Urology & Nephrology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Peptides Or Proteins (AREA)

Abstract

La présente invention concerne un anticorps anti-Siglec-15 et son utilisation. L'anticorps anti-Siglec-15 se lie spécifiquement à la protéine Siglec-15 humaine avec une valeur KD inférieure ou égale à environ 4E-09M. La présente invention concerne également un immunoconjugué comprenant l'anticorps anti-Siglec-15, un procédé de préparation de l'anticorps anti-Siglec-15, et l'utilisation de l'anticorps anti-Siglec-15.
PCT/CN2022/088241 2021-04-22 2022-04-21 Anticorps anti-siglec-15 et son utilisation WO2022223004A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202280029885.0A CN117203241A (zh) 2021-04-22 2022-04-21 抗Siglec15抗体及其用途

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202110435142.0 2021-04-22
CN202110435142 2021-04-22

Publications (1)

Publication Number Publication Date
WO2022223004A1 true WO2022223004A1 (fr) 2022-10-27

Family

ID=83723565

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/088241 WO2022223004A1 (fr) 2021-04-22 2022-04-21 Anticorps anti-siglec-15 et son utilisation

Country Status (2)

Country Link
CN (1) CN117203241A (fr)
WO (1) WO2022223004A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024040195A1 (fr) 2022-08-17 2024-02-22 Capstan Therapeutics, Inc. Conditionnement pour l'ingénierie de cellules immunitaires in vivo

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102803293A (zh) * 2009-10-06 2012-11-28 阿莱斯亚生物疗法股份有限公司 治疗骨丢失相关疾病的siglec 15抗体
CN104321430A (zh) * 2012-03-30 2015-01-28 第一三共株式会社 新颖的抗-siglec-15抗体

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102803293A (zh) * 2009-10-06 2012-11-28 阿莱斯亚生物疗法股份有限公司 治疗骨丢失相关疾病的siglec 15抗体
CN104321430A (zh) * 2012-03-30 2015-01-28 第一三共株式会社 新颖的抗-siglec-15抗体

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024040195A1 (fr) 2022-08-17 2024-02-22 Capstan Therapeutics, Inc. Conditionnement pour l'ingénierie de cellules immunitaires in vivo
WO2024040194A1 (fr) 2022-08-17 2024-02-22 Capstan Therapeutics, Inc. Conditionnement pour l'ingénierie de cellules immunitaires in vivo

Also Published As

Publication number Publication date
CN117203241A (zh) 2023-12-08

Similar Documents

Publication Publication Date Title
KR102340832B1 (ko) 항 pd-1 항체 및 그의 용도
KR102365972B1 (ko) 항-pd-1 항체 및 이의 용도
US9205148B2 (en) Antibodies and other molecules that bind B7-H1 and PD-1
US11525005B2 (en) Anti-CD40 antibody, antigen binding fragment thereof and medical use thereof
CN113286634A (zh) 对gucy2c特异性的抗体及其用途
KR20200061320A (ko) 세포독성 t-림프구-관련 단백질 4 (ctla-4)에 대한 신규의 단일클론 항체
AU2013363962B2 (en) Anti-H7CR antibodies
TWI745670B (zh) 抗cd27抗體、其抗原結合片段及其醫藥用途
KR20220062500A (ko) 항-cd39 항체 조성물 및 방법
EP3892634A1 (fr) Anticorps anti-cd40, fragment de liaison à l'antigène de celui-ci et utilisation pharmaceutique associée
US20230242660A1 (en) Combination therapy involving anti-cd39 antibodies and anti-pd-1 or anti-pd-l1 antibodies
WO2021063352A1 (fr) Protéine de liaison à l'antigène anti-pd-l1 et son application
CN112424358A (zh) 与细胞粘附分子3结合的抗体
WO2022223004A1 (fr) Anticorps anti-siglec-15 et son utilisation
KR20160072268A (ko) 항-efna4 항체-약물 접합체
CN116178545A (zh) 一种抗人pd-l1人源化抗体或其抗原结合片段及其应用
WO2021000813A1 (fr) Anticorps monoclonal ciblant pd-1 et son utilisation
JP7014783B2 (ja) 抗ｃｄ２７抗体、その抗原結合性フラグメント、およびそのものの医学的使用
CN112969715A (zh) 一种抗cd47抗原结合蛋白及其应用
CN115521378B (zh) Pd-l1抗体及其用途
WO2023030311A1 (fr) Protéine de liaison à l'antigène ciblant siglec15 et son utilisation
WO2023284741A1 (fr) Anticorps anti-pd-1 et son utilisation
RU2779128C2 (ru) Антитело к cd40, его антигенсвязывающий фрагмент и его медицинское применение

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22791110

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 202280029885.0

Country of ref document: CN

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 22791110

Country of ref document: EP

Kind code of ref document: A1