WO2019154315A1 - 抗b7-h4抗体、其抗原结合片段及其医药用途 - Google Patents

抗b7-h4抗体、其抗原结合片段及其医药用途 Download PDF

Info

Publication number
WO2019154315A1
WO2019154315A1 PCT/CN2019/074397 CN2019074397W WO2019154315A1 WO 2019154315 A1 WO2019154315 A1 WO 2019154315A1 CN 2019074397 W CN2019074397 W CN 2019074397W WO 2019154315 A1 WO2019154315 A1 WO 2019154315A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
antibody
variable region
chain variable
antigen
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/CN2019/074397
Other languages
English (en)
French (fr)
Chinese (zh)
Inventor
包如迪
花海清
刘素霞
张福军
王婷
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangsu Hansoh Pharmaceutical Group Co Ltd
Shanghai Hansoh Biomedical Co Ltd
Original Assignee
Jiangsu Hansoh Pharmaceutical Group Co Ltd
Shanghai Hansoh Biomedical Co Ltd
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
Priority to CA3089246A priority Critical patent/CA3089246A1/en
Priority to AU2019218319A priority patent/AU2019218319B2/en
Priority to BR112020015736-8A priority patent/BR112020015736A2/pt
Priority to RU2020124155A priority patent/RU2792748C2/ru
Priority to CN201980001361.9A priority patent/CN110366560B/zh
Priority to KR1020207026012A priority patent/KR102952441B1/ko
Priority to EP19750949.0A priority patent/EP3753951A4/en
Priority to JP2020542138A priority patent/JP7393337B2/ja
Priority to CN202310820681.5A priority patent/CN116693686A/zh
Priority to US16/967,016 priority patent/US11472882B2/en
Application filed by Jiangsu Hansoh Pharmaceutical Group Co Ltd, Shanghai Hansoh Biomedical Co Ltd filed Critical Jiangsu Hansoh Pharmaceutical Group Co Ltd
Priority to MX2020008181A priority patent/MX2020008181A/es
Publication of WO2019154315A1 publication Critical patent/WO2019154315A1/zh
Priority to ZA2020/04701A priority patent/ZA202004701B/en
Anticipated expiration legal-status Critical
Priority to JP2023197481A priority patent/JP2024026132A/ja
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2827—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against B7 molecules, e.g. CD80, CD86
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • 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
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/51—Complete heavy chain or Fd fragment, i.e. VH + CH1
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/515—Complete light chain, i.e. VL + CL
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/74—Inducing cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present invention relates to an anti-B7-H4 antibody immunoreactive to a human B7-H4 receptor, and an antigen-binding fragment thereof, a chimeric antibody, a humanized antibody comprising the CDR region of the anti-B7-H4 antibody, and A pharmaceutical composition comprising a human anti-B7-H4 antibody and an antigen-binding fragment thereof, and use thereof as an anticancer drug.
  • Tumor immunotherapy is a long-term research and development hotspot in the field of cancer treatment, and T cell tumor immunotherapy is at the core. Tumor escape is a huge obstacle to tumor immunotherapy. Most tumor expression can be recognized by the host immune system to varying degrees, but in many cases, due to inefficient activation of effector T cells, an inadequate immune response is triggered. Tumor growth is promoted by its own inhibition of the immune system. Tumor immunotherapy is to fully utilize and mobilize killer T cells and/or other immune cells in tumor patients to kill tumors.
  • B7 superfamily a class of immunoglobulins with an immunoglobulin V-like domain (IgV) and an immunoglobulin C-like domain (IgC), members of which include costimulatory factors B7.1 (CD80) and B7.2 ( CD86), inducible ligand for stimulatory factor (ICOS-L/B7-H2), programmed death-1 ligand (PD-L1/B7-H1), programmed death-2 ligand (PD- L2/B7-DC), B7-H4 and B7-H4, etc.
  • IgV immunoglobulin V-like domain
  • IgC immunoglobulin C-like domain
  • Human B7-H4 is a type I transmembrane protein consisting of 282 amino acids whose coding gene is located in the p11.1 region of chromosome 1 (Choi IH et al., J Immunol. 2003 Nov 1; 171(9): 4650-4) .
  • B7-H4 plays a negative role in the regulation of T cell immune response.
  • B7-H4 plays a wide-ranging role in the differentiation and development of CD4 + and CD8 + T cells, cell cycle progression and cytokine production (Sica GL, etc.) , Immunity. 2003 Jun; 18(6): 849-61).
  • No immune cell disorders and autoimmune phenomena were found in B7-H4 knockout mice (Zhu G et al, Blood. 2009 Feb 19; 113(8): 1759-67; Suh WK et al., Blood. Mol Cell Biol. 2006 Sep;26(17):6403-11).
  • the receptors for B7-H4 and their signal transduction pathways are still
  • B7-H4 protein is abundantly expressed in various tumor tissues, allowing tumor cells to escape the attack of the body's immune system.
  • the use of B7-H4 molecule as a tumor therapeutic target provides a new method for tumor immunotherapy.
  • B7-H4 is expressed on various cancer cells such as breast cancer, ovarian cancer, lung cancer, cervical cancer, kidney cancer, bladder cancer and liver cancer.
  • B7-H4 mRNA expression was found in the spleen, lung, thymus, liver, skeletal muscle, kidney, pancreas, testis and ovary.
  • At the protein level there were low tissues in the breast (catheter and lobular), fallopian tube epithelium, endometrial gland, etc. Horizontal B7-H4 expression.
  • TAM tumor-associated macrophages
  • macrophages constitute An important component of the tumor microenvironment and can represent up to 50% of tumor mass.
  • the present invention provides anti-B7-H4 antibodies with high affinity, high selectivity and high biological activity, monoclonal antibody immunotherapy for tumors and related applications. Drugs, compositions, and methods for the treatment of B7-H4 positive tumors.
  • the present invention provides a B7-H4 antibody or antigen-binding fragment thereof, comprising:
  • An antibody light chain variable region comprising at least one LCDR selected from the group consisting of:
  • SEQ ID NO: 14 SEQ ID NO: 15, SEQ ID NO: 16;
  • SEQ ID NO: 62 SEQ ID NO: 63, SEQ ID NO: 64;
  • An antibody heavy chain variable region comprising at least one HCDR selected from the group consisting of:
  • SEQ ID NO: 3 SEQ ID NO: 4, SEQ ID NO: 5;
  • SEQ ID NO: 51 SEQ ID NO: 52, SEQ ID NO: 53;
  • an anti-B7-H4 antibody or antigen-binding fragment thereof, as described above, wherein said antibody heavy chain variable region comprises:
  • HCDR1, HCDR2 and HCDR3 set forth in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5;
  • HCDR1, HCDR2 and HCDR3 represented by SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21;
  • HCDR1, HCDR2 and HCDR3 set forth in SEQ ID NO:27, SEQ ID NO:28 and SEQ ID NO:29;
  • HCDR1, HCDR2 and HCDR3 set forth in SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37;
  • HCDR1, HCDR2 and HCDR3 set forth in SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45;
  • HCDR1, HCDR2 and HCDR3 represented by SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53;
  • HCDR1, HCDR2 and HCDR3 represented by SEQ ID NO: 59, SEQ ID NO: 60 and SEQ ID NO: 61;
  • HCDR1, HCDR2 and HCDR3 represented by SEQ ID NO: 67, SEQ ID NO: 68 and SEQ ID NO: 69;
  • HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:73, SEQ ID NO:74 and SEQ ID NO:21.
  • an anti-B7-H4 antibody or antigen-binding fragment thereof, wherein said antibody light chain variable region comprises:
  • LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8;
  • LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16;
  • LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24;
  • LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32;
  • LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 38, SEQ ID NO: 39 and SEQ ID NO: 40;
  • LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48;
  • LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56;
  • a particularly preferred anti-B7-H4 antibody or antigen-binding fragment thereof may be selected from any one of the group consisting of one or more CDR region sequences selected from or having at least 95% sequence identity to:
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively; the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16, respectively;
  • the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13, respectively.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24, respectively; the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32, respectively; the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:27, SEQ ID NO:28 and SEQ ID NO:29.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 38, SEQ ID NO: 39 and SEQ ID NO: 40, respectively;
  • the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively; the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively;
  • the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 51, SEQ ID NO: 52 and SEQ ID NO: 53, respectively.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 62, SEQ ID NO: 63 and SEQ ID NO: 64, respectively; the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 59, SEQ ID NO: 60 and SEQ ID NO: 61, respectively.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 70, SEQ ID NO: 71 and SEQ ID NO: 72, respectively; the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 67, SEQ ID NO: 68 and SEQ ID NO: 69, respectively.
  • the antibody light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24, respectively; the antibody heavy chain variable region comprises For example, HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 73, SEQ ID NO: 74 and SEQ ID NO: 21.
  • a B7-H4 antibody or antigen-binding fragment thereof wherein the antibody or antigen-binding fragment thereof is a murine antibody or a fragment thereof.
  • a B7-H4 antibody or antigen-binding fragment thereof wherein the antibody or antigen-binding fragment thereof is a chimeric antibody or a fragment thereof.
  • the anti-B7-H4 antibody or antigen-binding fragment thereof wherein the antibody or antigen-binding fragment thereof is a human antibody or a fragment thereof.
  • a B7-H4 antibody or antigen-binding fragment thereof wherein the antibody or antigen-binding fragment thereof is a humanized antibody or a fragment thereof.
  • a B7-H4 antibody or antigen-binding fragment thereof, wherein the humanized antibody light chain variable region is selected from the group consisting of: SEQ ID NO:76, SEQ ID NO:78, SEQ ID NO:80 or SEQ ID NO:82.
  • a B7-H4 antibody or antigen-binding fragment thereof wherein said humanized antibody heavy chain variable region is selected from the group consisting of: SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79 or SEQ ID NO:81.
  • a B7-H4 antibody or antigen-binding fragment thereof wherein the humanized antibody heavy chain variable region further comprises human IgG1, IgG2, IgG3 or IgG4 or
  • the heavy chain FR region of the variant preferably comprises a human IgGl, IgG2 or IgG4 heavy chain FR region; more preferably IgGl which enhances ADCC toxicity following amino acid mutations.
  • a B7-H4 antibody or antigen-binding fragment thereof, wherein the light chain of the humanized antibody is selected from the group consisting of: SEQ ID NO: 84 SEQ ID NO:86, SEQ ID NO:88 or SEQ ID NO:90.
  • a B7-H4 antibody or antigen-binding fragment thereof, wherein the heavy chain of the humanized antibody is selected from the group consisting of: SEQ ID NO: 83 SEQ ID NO:85, SEQ ID NO:87 or SEQ ID NO:89.
  • a B7-H4 antibody or antigen-binding fragment thereof, wherein the humanized antibody light chain variable region is selected from the group consisting of: SEQ ID NO: 76 or SEQ ID NO: 80.
  • a B7-H4 antibody or antigen-binding fragment thereof, wherein the light chain of the humanized antibody is selected from the group consisting of: SEQ ID NO: 84 Or SEQ ID NO:88.
  • a B7-H4 antibody or antigen-binding fragment thereof, wherein the heavy chain of the humanized antibody is selected from the group consisting of: SEQ ID NO: 83 Or SEQ ID NO:87.
  • the humanized antibody is selected from any of the following:
  • the humanized antibody is selected from any one of the following:
  • An anti-B7-H4 antibody or antigen-binding fragment thereof having at least one of the following characteristics: (1) binding to an epitope of amino acids 41-60 of SEQ ID NO: 92 in B7-H4; (2) binding to B7 The epitope of amino acids 53-59 of SEQ ID NO: 92 is contained in -H4.
  • An anti-B7-H4 antibody or antigen-binding fragment thereof having at least one of the following characteristics: (1) binding to an epitope of amino acid 53 of SEQ ID NO: 92 in B7-H4; (2) binding to B7-H4 An epitope comprising amino acid 54 of SEQ ID NO: 92; (3) binding to an epitope of amino acid 56 of SEQ ID NO: 92 in B7-H4; (4) binding to B7-H4 containing SEQ ID NO: 92 Epitope of amino acid 57; (5) binding to an epitope of amino acid 58 of SEQ ID NO: 92 in B7-H4; (2) binding to an epitope of amino acid 59 of SEQ ID NO: 92 in B7-H4.
  • a B7-H4 antibody or antigen-binding fragment thereof wherein the antigen-binding fragment is Fab, Fv, sFv, F(ab') 2 , a linear antibody, Single-chain antibodies, Nanobodies, domain antibodies, and multispecific antibodies.
  • the present invention further provides a DNA sequence encoding the B7-H4 antibody or antigen-binding fragment thereof as described above.
  • the invention further provides an expression vector comprising the DNA sequence as described above.
  • the invention further provides a host cell introduced or containing an expression vector as described above.
  • a host cell as described above characterized in that said host cell is a bacterium, preferably Escherichia coli.
  • a host cell as described above is a yeast, preferably Pichia pastoris.
  • a host cell as described above is a mammalian cell, preferably a Chinese hamster ovary (CHO) cell or a human embryonic kidney (HEK) 293 cell.
  • CHO Chinese hamster ovary
  • HEK human embryonic kidney
  • the invention also provides a method of producing a B7-H4 antibody comprising: culturing the host cell as described above, isolating the antibody from the culture, and purifying the antibody.
  • the invention also provides a multispecific antibody comprising a light chain variable region and a heavy chain variable region as previously described.
  • the invention also provides a single chain antibody comprising a light chain variable region and a heavy chain variable region as previously described.
  • the invention also provides a detection reagent or diagnostic agent comprising a B7-H4 antibody or antigen-binding fragment thereof as described above.
  • the present invention also provides a method for immunodetection or assay of B7-H4, which comprises using the B7-H4 antibody of the present invention or an antigen-binding fragment thereof.
  • the present invention also provides a method for diagnosing a disease associated with a B7-H4 positive cell, the method comprising detecting or measuring B7-H4 or B7-H4 using the B7-H4 antibody or antigen-binding fragment thereof of the present invention. Positive cells.
  • the invention further provides a pharmaceutical composition
  • a pharmaceutical composition comprising a B7-H4 antibody or antigen-binding fragment thereof as described above and a pharmaceutically acceptable excipient, dilution or carrier.
  • the invention further provides the use of an anti-B7-H4 antibody or antigen-binding fragment thereof as described above, in the manufacture of a medicament for the treatment of a B7-H4 mediated disease or condition; wherein the disease is preferably cancer;
  • the cancer is B7-H4; the cancer is most preferably breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, lung cancer, liver cancer, stomach cancer, colon cancer, bladder cancer, esophageal cancer, cervical cancer, gallbladder Cancer, glioblastoma and melanoma.
  • the invention further provides a method of treating and preventing a B7-H4 mediated disease or condition, the method comprising administering to a subject in need thereof a therapeutically effective amount of an anti-B7-H4 antibody or antigen-binding fragment thereof, or a pharmaceutical combination thereof
  • the disease wherein the disease is preferably cancer; more preferably a cancer expressing B7-H4; the cancer is most preferably breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, lung cancer, liver cancer, stomach cancer, colon Cancer, bladder cancer, esophageal cancer, gallbladder cancer, cervical cancer, glioblastoma and melanoma.
  • Figure 1 In vitro ELISA binding experiments antibodies, chimeric antibodies Showing 7 binding activity of human B7-H4 purified antigen, wherein the chimeric antibody 2F7 and 2F8 EC 50 of approximately 0.1nM.
  • Figure 2 Indirect ELISA binding experiments showing antigenic epitopes of anti-B7-H4 antibody hu2F7 binding.
  • Figure 4 Immunological function experiments showing the effect of enhanced immunity (T cell proliferation) of anti-B7-H4 antibodies hu1C9 and hu2G6.
  • antibody refers to an immunoglobulin which is a tetrapeptide chain structure in which two identical heavy chains and two identical light chains are linked by interchain disulfide bonds.
  • the immunoglobulin heavy chain constant region has different amino acid composition and arrangement order, so its antigenicity is also different. Accordingly, immunoglobulins can be classified into five classes, or isoforms of immunoglobulins, namely IgM, IgD, IgG, IgA and IgE, and the corresponding heavy chains are ⁇ chain, ⁇ chain, ⁇ chain, respectively. , ⁇ chain and ⁇ chain.
  • IgG can be classified into IgG1, IgG2, IgG3, and IgG4.
  • Light chains are classified as either a kappa chain or a lambda chain by the constant region.
  • Each of the five types of Ig may have a kappa chain or a lambda chain.
  • the antibody light chain variable region of the present invention may further comprise a light chain constant region comprising a human or murine kappa, lambda chain or a variant thereof.
  • the antibody heavy chain variable region of the present invention may further comprise a heavy chain constant region comprising human or murine IgG1, 2, 3, 4 or a variant thereof.
  • variable region The sequence of about 110 amino acids near the N-terminus of the antibody heavy and light chains varies greatly, being the variable region (V region); the remaining amino acid sequence near the C-terminus is relatively stable and is a constant region (C region).
  • the variable region includes three hypervariable regions (HVR) and four relatively conserved framework regions (FR). The three hypervariable regions determine the specificity of the antibody, also known as the complementarity determining region (CDR).
  • Each of the light chain variable region (VL) and the heavy chain variable region (VH) consists of three CDR regions and four FR regions, and the order from the amino terminus to the carboxy terminus is: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • the three CDR regions of the light chain refer to LCDR1, LCDR2, and LCDR3; the three CDR regions of the heavy chain refer to HCDR1, HCDR2, and HCDR3.
  • the CDR amino acid residues of the VL and VH regions of the antibodies or antigen-binding fragments of the invention conform to known Kabat numbering rules and Kabat or AbM definition rules in number and position (http://bioinf.org.uk/abs/ ).
  • APC antigen presenting cell
  • T cells recognize this complex using the T cell receptor (TCR).
  • APCs include, but are not limited to, dendritic cells (DC), peripheral blood mononuclear cells (PBMC), monocytes, B lymphoblasts, and monocyte-derived dendritic cells (DC).
  • DC dendritic cells
  • PBMC peripheral blood mononuclear cells
  • monocytes B lymphoblasts
  • DC monocyte-derived dendritic cells
  • the term “antigen presentation” refers to the process by which APCs capture antigens and enable them to be recognized by T cells, for example as a component of MHC-I/MHC-II conjugates.
  • B7-H4 refers to a member of the human B7 protein family, also known as CD276, which is a type I transmembrane protein with four Ig-like extracellular domains.
  • B7-H4 is one of the immunological checkpoint proteins expressed on the surface of antigen-presenting cells or cancer cells, and has an inhibitory effect on the functional activation of T cells.
  • B7-H4 includes any variant or isoform of B7-H4 naturally expressed by a cell.
  • the antibodies of the invention can be cross-reactive with B7-H4 from non-human species. Alternatively, the antibody may also be specific for human B7-H4 and may not exhibit cross-reactivity with other species.
  • B7-H4 or any variant or isoform thereof, can be isolated from cells or tissues in which they are naturally expressed, or produced by recombinant techniques using techniques common in the art and described herein.
  • the anti-B7-H4 antibody targets human B7-H4 with a normal glycosylation pattern.
  • recombinant human antibody includes human antibodies which are prepared, expressed, created or isolated by recombinant methods, and the techniques and methods involved are well known in the art, such as (1) transgenes from human immunoglobulin genes, transgenic chromosomes.
  • Such recombinant human antibodies comprise variable regions and constant regions that utilize specific human germline immunoglobulin sequences encoded by germline genes, but also include subsequent rearrangements and mutations such as occur during antibody maturation.
  • murine antibody is in the present invention a monoclonal antibody to human B7-H4 prepared according to the knowledge and skill in the art.
  • the test subject is injected with the B7-H4 antigen at the time of preparation, and then the hybridoma expressing the antibody having the desired sequence or functional properties is isolated.
  • the murine B7-H4 antibody or antigen-binding fragment thereof may further comprise a light chain constant region of a murine kappa, a lambda chain or a variant thereof, or further comprising a murine IgG1 , heavy chain constant region of IgG2, IgG3 or IgG4 or a variant thereof.
  • human antibody includes antibodies having variable and constant regions of human germline immunoglobulin sequences.
  • Human antibodies of the invention can include amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo).
  • human antibody does not include an antibody in which a CDR sequence derived from a germline of another mammalian species, such as a mouse, has been grafted onto a human framework sequence (ie, "humanized antibody”).
  • humanized antibody also referred to as a CDR-grafted antibody, refers to an antibody produced by grafting a CDR sequence of a mouse into a human antibody variable region framework. It is possible to overcome the strong immune response induced by chimeric antibodies by carrying a large amount of mouse protein components. To avoid a decrease in activity while reducing immunogenicity, the human antibody variable region can be subjected to minimal reverse mutation to maintain activity.
  • chimeric antibody is an antibody obtained by fusing a variable region of a murine antibody with a constant region of a human antibody, and can alleviate an immune response induced by a murine antibody.
  • a hybridoma that secretes a murine-specific monoclonal antibody is selected, and then the variable region gene is cloned from the mouse hybridoma cell, and the constant region gene of the human antibody is cloned as needed to change the mouse.
  • the region gene and the human constant region gene are ligated into a chimeric gene and inserted into a human vector, and finally the chimeric antibody molecule is expressed in a eukaryotic industrial system or a prokaryotic industrial system.
  • the constant region of a human antibody may be selected from the heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4 or variants thereof, preferably comprising a human IgG2 or IgG4 heavy chain constant region, or enhanced ADCC using an amino acid mutation (antibody-dependent) Cell-mediated cytotoxicity, antibody-dependent cell-mediated cytotoxicity) toxic IgG1.
  • antigen-binding fragment refers to an antigen-binding fragment of an antibody and an antibody analog, which typically includes at least a portion of an antigen binding or variable region (eg, one or more CDRs) of a parental antibody.
  • the antibody fragment retains at least some of the binding specificity of the parent antibody. Generally, when the activity is expressed on a molar basis, the antibody fragment retains at least 10% of the parental binding activity. Preferably, the antibody fragment retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the binding affinity of the parent antibody to the target.
  • antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, linear antibodies, single-chain antibodies, Nanobodies, domain antibodies, and multispecific antibodies.
  • Engineered antibody variants are reviewed in Holliger and Hudson (2005) Nat. Biotechnol. 23: 1126-1136.
  • a "Fab fragment” consists of a light chain and a heavy chain CH1 and a variable region.
  • the heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule.
  • the "Fc" region contains two heavy chain fragments comprising the CH1 and CH2 domains of the antibody.
  • the two heavy chain fragments are held together by two or more disulfide bonds and by the hydrophobic action of the CH3 domain.
  • a "Fab' fragment” contains a light chain and a portion of a heavy chain comprising a VH domain and a CH1 domain and a region between the CH1 and CH2 domains, thereby being between the two heavy chains of the two Fab' fragments An interchain disulfide bond is formed to form a F(ab')2 molecule.
  • the "F(ab')2 fragment” contains two light chains and two heavy chains comprising a portion of the constant region between the CH1 and CH2 domains, thereby forming an interchain disulfide bond between the two heavy chains.
  • the F(ab')2 fragment consists of two Fab' fragments held together by a disulfide bond between the two heavy chains.
  • the "Fv region” contains variable regions from both heavy and light chains, but lacks a constant region.
  • multispecific antibody is used in its broadest sense to encompass antibodies having multi-epitope specificity.
  • These multispecific antibodies include, but are not limited to, antibodies comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH-VL unit has multi-epitope specificity; having two or more Antibodies of the VL and VH regions, each VH-VL unit binding to a different target or a different epitope of the same target; antibodies having two or more single variable regions, each single variable region Different targets or different epitopes of the same target; full-length antibodies, antibody fragments, diabodies, bispecific diabodies and triabodies, covalently or non-covalently linked Along with antibody fragments and the like.
  • single-chain antibody is a single-stranded recombinant protein joined by a heavy chain variable region (VH) and a light chain variable region (VL) of a antibody via a ligation peptide, which is the smallest with complete antigen binding sites.
  • VH heavy chain variable region
  • VL light chain variable region
  • domain antibody fragment is an immunologically functional immunoglobulin fragment containing only a heavy chain variable region or a light chain variable region chain.
  • two or more VH regions are covalently joined to a peptide linker to form a bivalent domain antibody fragment.
  • the two VH regions of a bivalent domain antibody fragment can target the same or different antigens.
  • binding to B7-H4" of the present invention means that it is capable of interacting with human B7-H4.
  • antigen binding site refers to a three-dimensional spatial site that is discrete on an antigen and is recognized by an antibody or antigen-binding fragment of the present invention.
  • epitope refers to a site on an antigen that specifically binds to an immunoglobulin or antibody.
  • An epitope can be formed by an adjacent amino acid, or a non-adjacent amino acid juxtaposed by a tertiary folding of a protein. Epitopes formed from adjacent amino acids are typically retained after exposure to denaturing solvents, while epitopes formed by tertiary folding are typically lost after treatment with denaturing solvents. Epitopes typically include at least 3-15 amino acids in a unique spatial conformation. Methods for determining what epitope is bound by a given antibody are well known in the art, including immunoblotting and immunoprecipitation assays, and the like. Methods for determining the spatial conformation of an epitope include techniques in the art and techniques described herein, such as X-ray crystallography and two-dimensional nuclear magnetic resonance.
  • the terms “specifically bind” and “selectively bind” refer to the binding of an antibody to an epitope on a predetermined antigen.
  • the antibody has an equilibrium solution of less than about 10 -7 M or even less when measured by surface plasmon resonance (SPR) techniques in an instrument.
  • the isolating constant (K D ) binds to a predetermined antigen, and its affinity for binding to a predetermined antigen is at least twice its affinity for binding to a non-specific antigen other than the predetermined antigen or a closely related antigen (such as BSA, etc.).
  • the term "antibody recognizing an antigen” can be used interchangeably herein with the term “specifically bound antibody”.
  • cross-reactive refers to the ability of an antibody of the invention to bind to B7-H4 from a different species.
  • an antibody of the invention that binds to human B7-H4 can also bind to B7-H4 of another species.
  • Cross-reactivity is measured by detecting specific reactivity with purified antigens in binding assays (eg, SPR and ELISA), or binding or functional interactions with cells that physiologically express B7-H4.
  • binding assays eg, SPR and ELISA
  • Methods for determining cross-reactivity include standard binding assays as described herein, such as surface plasmon resonance (SPR) analysis, or flow cytometry.
  • SPR surface plasmon resonance
  • inhibiting or “blocking” are used interchangeably and encompass both partial and complete inhibition/blocking.
  • the inhibition/blocking of the ligand preferably reduces or alters the normal level or type of activity that occurs when ligand binding occurs without inhibition or blockade.
  • Inhibition and blockade are also intended to include a decrease in any measurable ligand binding affinity when contacted with an anti-B7-H4 antibody, compared to a ligand not contacted with an anti-B7-H4 antibody.
  • inhibiting growth eg, involving cells
  • inhibiting growth is intended to include any measurable reduction in cell growth.
  • inducing an immune response and “enhancing an immune response” are used interchangeably and refer to the stimulation (ie, passive or adaptive) of an immune response to a particular antigen.
  • inducing for inducing CDC or ADCC refers to stimulating a specific direct cell killing mechanism.
  • the "ADCC” described in the present invention is an antibody-dependent cell-mediated cytotoxicity, which means that a cell expressing an Fc receptor directly kills an antibody by Fc segment of the recognition antibody.
  • Target cells The ADCC effector function of the antibody can be reduced or eliminated by modification of the Fc fragment on IgG. Said modification refers to mutations in the heavy chain constant region of the antibody.
  • a mouse can be immunized with human B7-H4 or a fragment thereof, and the obtained antibody can be renatured, purified, and subjected to amino acid sequencing by a conventional method.
  • the antigen-binding fragment can also be prepared by a conventional method.
  • the antibodies or antigen-binding fragments of the invention are genetically engineered to add one or more human FR regions in a non-human CDR region. Human FR germline sequences are available on the website of ImMunoGeneTics (IMGT) http://imgt.cines.fr or from the Journal of Immunoglobulins, 2001 ISBN 014441351.
  • the engineered antibodies or antigen-binding fragments of the invention can be prepared and purified by conventional methods.
  • the cDNA sequence of the corresponding antibody can be cloned and recombined into a GS expression vector.
  • the recombinant immunoglobulin expression vector can stably transfect CHO cells.
  • mammalian expression systems result in glycosylation of antibodies, particularly at the highly conserved N-terminus of the FC region.
  • Stable clones are obtained by expressing antibodies that specifically bind to human antigens. Positive clones were expanded in serum-free medium in a bioreactor to produce antibodies.
  • the culture medium from which the antibody is secreted can be purified and collected by a conventional technique.
  • the antibody can be concentrated by filtration in a conventional manner. Soluble mixtures and multimers can also be removed by conventional methods such as molecular sieves, ion exchange. The resulting product needs to be frozen immediately, such as -70 ° C, or lyophilized.
  • the antibody of the present invention refers to a monoclonal antibody.
  • the monoclonal antibody (mAb) of the present invention refers to an antibody obtained from a single clonal cell strain, and the cell strain is not limited to a eukaryotic, prokaryotic or phage clonal cell strain.
  • Monoclonal antibodies or antigen-binding fragments can be obtained recombinantly using, for example, hybridoma technology, recombinant techniques, phage display technology, synthetic techniques (e.g., CDR-grafting), or other prior art techniques.
  • administering when applied to an animal, human, experimental subject, cell, tissue, organ or biological fluid, refers to an exogenous drug, therapeutic agent, diagnostic agent or composition and animal, human, subject Contact of the test subject, cell, tissue, organ or biological fluid.
  • administering can refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods.
  • Treatment of the cells includes contact of the reagents with the cells, and contact of the reagents with the fluid, wherein the fluids are in contact with the cells.
  • administeristering and “treating” also means treating, for example, cells in vitro and ex vivo by reagents, diagnostics, binding compositions, or by another cell.
  • Treatment when applied to a human, veterinary or research subject, refers to therapeutic treatment, prophylactic or preventive measures, research and diagnostic applications.
  • Treatment means administering to a patient a therapeutic agent for internal or external use, such as a composition comprising any of the binding compounds of the present invention, the patient having one or more symptoms of the disease, and the therapeutic agent is known to have Therapeutic effect.
  • a therapeutic agent is administered in a subject or population to be treated to effectively alleviate the symptoms of one or more diseases, whether by inducing such symptoms to degenerate or inhibiting the progression of such symptoms to any degree of clinical right measurement.
  • the amount of therapeutic agent also referred to as "therapeutically effective amount” effective to alleviate the symptoms of any particular disease can vary depending on a variety of factors, such as the patient's disease state, age and weight, and the ability of the drug to produce a desired effect in the patient.
  • Whether the symptoms of the disease have been alleviated can be assessed by any clinical test method commonly used by a physician or other professional health care provider to assess the severity or progression of the condition.
  • Embodiments of the invention e.g., methods of treatment or preparations
  • a binding compound consisting essentially of the amino acid sequence recited may also include one or more amino acids that do not significantly affect the properties of the binding compound.
  • naturally occurring refers to the fact that the object can be found in nature.
  • a polypeptide sequence or polynucleotide sequence that is present in an organism (including viruses) that can be isolated from a natural source and that has not been intentionally modified in the laboratory by humans is naturally occurring.
  • an "effective amount” includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical condition.
  • An effective amount also means an amount sufficient to allow or facilitate the diagnosis.
  • An effective amount for a particular patient or veterinary subject can vary depending on factors such as the condition to be treated, the overall health of the patient, the route and dosage of the method of administration, and the severity of the side effects.
  • An effective amount can be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.
  • Exogenous refers to a substance that is produced outside of a living organism, cell, or human body depending on the background.
  • Endogenous refers to a substance produced in a cell, organism or human body depending on the background.
  • “Homology” refers to sequence similarity between two polynucleotide sequences or between two polypeptides. When positions in both comparison sequences are occupied by the same base or amino acid monomer subunit, for example if each position of two DNA molecules is occupied by adenine, then the molecule is homologous at that position .
  • the percent homology between the two sequences is a function of the number of matches or homology positions shared by the two sequences divided by the number of positions compared ⁇ 100%. For example, in the optimal alignment of sequences, if there are 6 matches or homologs in 10 positions in the two sequences, then the two sequences are 60% homologous. In general, comparisons are made when the maximum sequence of homology is obtained by aligning the two sequences.
  • “Pharmaceutical composition” means a mixture comprising one or more of the compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiological/pharmaceutically acceptable carriers. And excipients.
  • the purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient and thereby exerts biological activity.
  • the human B7-H4 (huB7-H4-HF) sequence encoding the HisFlag tag, and the human B7-H4 (h-B7-H4-Fc) sequence encoding the huFc tag were synthesized by CRO Integrated DNA Technology (IDT) (above B7)
  • the -H4 recombinant proteins were each designed from the template sequence of the present invention and cloned into the pTT5 vector (Biovector), respectively.
  • the recombinant B7-H4 protein was purified by Example 2 after expression in 293T cells.
  • the purified protein can be used in the experiments of the following examples.
  • the cell expression supernatant sample was centrifuged at high speed to remove impurities, and the buffer was exchanged for PBS, and imidazole was added to a final concentration of 5 mM.
  • the nickel column was equilibrated with PBS solution containing 5 mM imidazole and rinsed 2-5 column volumes. The displaced supernatant sample was applied to the column.
  • the column was washed with PBS containing 5 mM imidazole until the A280 reading dropped to baseline.
  • the column was washed with PBS + 10 mM imidazole, the non-specifically bound heteroprotein was removed, and the effluent was collected.
  • the protein of interest was eluted with PBS containing 300 mM imidazole, and the eluted peak was collected.
  • the collected eluate was further purified by ion exchange (SP column).
  • Configure solution A 0.01 M PB, pH 8.0.
  • Configure liquid B liquid A + 1 M NaCl.
  • the imidazole in PBS solution was eluted to the A solution, and the SP column was equilibrated with the A solution.
  • the concentration of the B solution was 0-100%, and 10 column volumes were eluted to collect the elution peaks.
  • the obtained protein was electrophoresed and identified as correct and then used.
  • the human B7-H4 (hu-B7-H4 his) with the HisFlag tag was obtained.
  • the supernatant sample expressed by HEK293 cells was centrifuged at high speed to remove impurities, and the buffer was exchanged for PBS.
  • the Protein A affinity column was equilibrated with 10 mM phosphate buffer and rinsed 2-5 column volumes. The displaced supernatant sample was applied to the column. Rinse the column with 25 column volumes of buffer until the A 280 reading drops to baseline.
  • the target protein was eluted with 0.8% acetate buffer at pH 3.5, and the elution peak was collected. Immediately after the addition, the mixture was neutralized with 1 M Tris-Cl pH 8.0 buffer, and then the solution was replaced with Millipore's Amico-15 filter column. .
  • the obtained protein was electrophoresed, peptide map, LC-MS identified and used.
  • the full-length sequence encoding the human or cynomolgus B7-H4 protein was synthesized by Integrated DNA Technology (IDT) (the above B7-H3 recombinant proteins were designed by the template of the present invention) and cloned into On the engineered pcDNA3.1 vector, pcDNA3.1/puro (Invitrogen #V79020).
  • CHO-S (ATCC) cells were cultured in the CD-CHO medium (Life Technologies, # 10743029) to 0.5x10 6 / ml.
  • the indirect ELISA, capture ELISA method as described in Example 3 was used to evaluate serum titer and the ability to bind cell surface antigens, and the control titer assay (greater than 100,000 fold dilution) was used to initiate cell fusion.
  • the mice were sacrificed after a final immunization with serum titer, affinity and FACS binding.
  • the spleen cells and SP2/0 myeloma cells were fused and plated to obtain hybridomas.
  • the target hybridization was screened by indirect ELISA and capture ELISA. Tumors were constructed and cloned into monoclonal cell lines by limiting dilution.
  • the obtained positive antibody strain was further subjected to CHO-S cells stably expressing B7-H4, and the blank CHO-S cells were compared to exclude non-specific antibody-binding hybridoma strains, and the cells were selected by flow sorting, thereby selecting 8 strains of binding.
  • Hybridoma cells in logarithmic growth phase were collected, RNA was extracted with Trizol (Invitrogen, 15596-018) and reverse transcription (PrimeScript TM Reverse Transcriptase, Takara # 2680A).
  • the cDNA obtained by reverse transcription was subjected to PCR amplification using a mouse Ig-Primer Set (Novagen, TB326 Rev. B 0503) and sequenced, and finally a sequence of 8 murine antibody was obtained.
  • the heavy and light chain variable region sequences of murine mAb 2F7 are as follows:
  • the heavy and light chain variable region sequences of M1 are as follows:
  • the heavy and light chain variable region sequences of murine mAb 2F8 are as follows:
  • the heavy and light chain variable region sequences of murine mAb 2F4 are as follows:
  • the heavy and light chain variable region sequences of murine mAb 2A10 are as follows:
  • the heavy and light chain variable region sequences of murine mAb 2E4 are as follows:
  • the heavy and light chain variable region sequences of murine mAb 1E4 are as follows:
  • the heavy and light chain variable region sequences of murine mAb 2G6 are as follows:
  • the heavy and light chain variable region sequences of murine mAb 1C9 are as follows:
  • the heavy and light chain variable regions of each mouse antiserum were separately cloned into the human IgG1 heavy chain constant region and the kappa light chain constant region, purified, identified, and tested for activity as described in Example 4.
  • the huB7-H4 His protein (Sino Biological Inc., cat#10738-H08H) was diluted to a concentration of 1 ⁇ g/ml with PBS of pH 7.4, and added to a 96-well high-affinity microtiter plate at a volume of 100 ⁇ l/well at 4 °C. Incubate overnight in the refrigerator (16-20 hours). After washing the plate 4 times with PBST (pH 7.4 PBS containing 0.05% Tween-20), 150 ⁇ l/well of a 3% bovine serum albumin (BSA) blocking solution diluted with PBST was added, and the mixture was incubated at room temperature for 1 hour to block. After the end of the blocking, the blocking solution was discarded and the plate was washed 4 times with PBST buffer.
  • BSA bovine serum albumin
  • the antibody to be tested was diluted with PBS containing 3% BSA, starting at 1 ⁇ M, 10 fold gradient, 10 doses, added to the plate at 100 ⁇ l/well, and incubated for 1 hour at room temperature. After the end of the incubation, the plate was washed 4 times with PBST, and 100 ⁇ l/well of HRP-labeled goat anti-human secondary antibody (Abeam, cat#ab97225) diluted with 3% BSA in PBST was added, and incubated for 1 hour at room temperature.
  • the huB7-H4His protein (Sino Biological Inc., cat#10738-H08H) was diluted to a concentration of 1 ⁇ g/ml with PBS of pH 7.4, and added to a 96-well high-affinity microtiter plate at a volume of 100 ⁇ l/well in a refrigerator at 4 ° C. Incubate overnight (16-20 hours), wash the plate 4 times with PBST (pH 7.4 PBS containing 0.05% Tween-20), add 150 ⁇ l/well of 3% bovine serum albumin (BSA) blocking solution diluted with PBST, incubate at room temperature. Closed for 1 hour. After the end of the blocking, the blocking solution was discarded and the plate was washed 4 times with PBST buffer.
  • BSA bovine serum albumin
  • 0.1 nM of the reference chimeric antibody was prepared with PBS containing 3% BSA, and the murine antibody to be tested was diluted to 100 nM with the antibody as a dilution, 10 fold gradient, 10 doses, and the diluted antibody was 100 ⁇ l/ The wells were added to the plate and incubated for 1 hour at room temperature. After the end of the incubation, the plate was washed 4 times with PBST, and 100 ⁇ l/well of HRP-labeled goat anti-human secondary antibody (Abeam, cat#ab97225) diluted with 3% BSA in PBST was added, and incubated for 1 hour at room temperature.
  • HRP-labeled goat anti-human secondary antibody Abeam, cat#ab97225
  • the goat anti-mouse IgG secondary antibody (Jackson Immuno Research, cat #115-006-071) was diluted to a concentration of 2 ug/ml with PBS buffer pH 7.4, and added to the 96-well microtiter plate at a volume of 100 ul/well. Place in the incubator at 37 ° C for 2 hours. After washing the plate once with PBST, a 5% skim milk (bright skim milk powder) blocking solution diluted with PBST was added at 200 ul/well, and incubated at 37 ° C for 2 hours or at 4 ° C overnight (16-18 hours) for blocking. After the end of the closure, the blocking solution was discarded and the plate was washed 4 times with PBST.
  • the plate was washed 4 times with PBST, and 100 ul/well of HRP-labeled streptavidin diluted with PBST (Jackson Immuno Research, cat #016-030-084) was added, and incubated at 37 ° C for 40 minutes. After washing the plate 4 times with PBST, add 100 ⁇ l/well TMB chromogenic substrate (Huzhou Yingchuang Biotechnology Co., Ltd.), incubate at room temperature for 10-15 min in the dark, add 50 ⁇ l/well 1M H 2 SO 4 to stop the reaction, and use the enzyme label.
  • the instrument (Beijing Pulang New Technology Co., Ltd., model DNM-9602) reads the absorption value at 450 nm and analyzes the data.
  • the cultured CHO-huB7-H4 stably transfected cells or SK-BR3 cells were collected, adjusted for cell density, and then plated in a 96-well U-bottom plate, 1 to 2 x 105 cells per well. Centrifuge at 1200g, 5min, remove the supernatant, add 100ul of the diluted antibody solution or mouse immune serum, incubate at 4 °C for 60min; 1200g, centrifuge for 5min, remove the supernatant, wash the cells twice with PBS, add fluorescent labeled secondary antibody (PE-GAM or PE-GAH) 100 ul per well, incubate for 60 min at 4 °C. 1200 g, centrifuged to remove the supernatant at 5 min. After washing the cells twice with PBS, they were resuspended in PBS, and the signals were detected using a flow cytometer, and the results of concentration curve analysis were performed.
  • PE-GAM or PE-GAH fluorescent labeled secondary antibody
  • the murine antibody heavy chain variable region (VH) and the light chain variable region (VL) of the present invention are respectively designated as site-directed amino acid mutations in the FR (framework region) region, and different people are designed according to different amino acid mutation combinations.
  • Humanized antibodies can be produced by transfecting the antibody heavy and light chains and transfecting different light and heavy chain combination plasmids into cells.
  • the heavy chain vector was designed as follows: signal peptide + mutated heavy chain variable region sequence + human IgGl constant region sequence.
  • the light chain vector is designed as follows: signal peptide + mutated light chain variable region sequence + human Kappa constant region sequence.
  • the above sequences were inserted into the pCEP4 vector, respectively.
  • the expression vector was synthesized according to the above design, and after obtaining the vector plasmid, the plasmid was greatly extracted, and the plasmid was subjected to sequencing verification.
  • the validated plasmids were transfected into human 293F cells with PEI, cultured continuously, and 293F cells were cultured in serum-free medium (Shanghai Aupmaxi, OPM-293CD03) to logarithmic growth phase for cell transfection.
  • the centrifuged cell culture solution was applied to an antibody purification affinity column, and washed with a phosphate buffer column, glycine hydrochloride buffer (pH 2.7 0.1 M Gly-HCl), 1 M Tris hydrochloric acid pH 9.0, and The phosphate buffer was dialyzed to finally obtain purified chimeric antibodies.
  • the Biacore method is recognized as a method for the objective detection of the affinity and kinetics of proteins.
  • the affinities and binding kinetics of the B7-H4 antibodies of the invention to be tested were analyzed by Biacore T200 (GE).
  • the anti-B7-H4 antibody of the present invention to be tested was covalently linked to a CM5 (GE) chip by a NHS standard amino coupling method using a kit supplied by Biacore. then:
  • Humanization of murine anti-human B7-H4 monoclonal antibodies was performed as disclosed in many literatures in the art. Briefly, the human constant domain is used in place of the parental (murine antibody) constant domain, and the human antibody sequence is selected based on the homology of the murine antibody and the human antibody.
  • the present invention provides the murine candidate molecules 2F7, 2F8, 2G6 and 1C9 is humanized.
  • the heavy and light chain variable region sequences were compared with the human antibody germline database to obtain a human germline template with high homology.
  • the CDR regions of the murine antibodies 2F7, 2F8, 2G6 and 1C9 were grafted onto the selected corresponding humanized template.
  • the HFR1 region of 2F8 (GYTFTNSWMN, SEQ ID NO: 19) and the HCDR2 region (GIYPNSGNIEYNEKFKG, SEQ ID NO: 20) were mutated to GYTFTSSWMN (SEQ ID NO: 73) and GIYPNRGNIEYNEKFKG (SEQ ID NO: 74), respectively, to remove potential Deacetylation unstable sites.
  • the humanized variable region is replaced and recombined with an IgG constant region (preferably the heavy chain is IgG1 and the light chain is ⁇ ).
  • the residues which have an important interaction with the CDRs and the residues of the CDRs, and the residues which have an important influence on the conformation of VL and VH are subjected to back mutation and the CDR regions are Optimization of chemically labile amino acid residues, designing and detecting antibodies resulting from the combination of humanized light and heavy chain variable region sequences.
  • the final humanized hu2F7, hu2F8, hu2G6 and hu1C9 antibody molecules were selected by expression test and the number of back mutations, and the respective heavy and light chain variable region sequences are shown in SEQ ID NOs: 75-82, each of which The heavy and light chain sequences are set forth in SEQ ID NOs: 83-90.
  • a cDNA fragment was synthesized based on the amino acid sequences of the above humanized antibody light and heavy chains, and inserted into a pcDNA3.1 expression vector (Life Technologies Cat. No. V790-20).
  • the expression vector and the transfection reagent PEI (Polysciences, Inc. Cat. No. 23966) were transfected into HEK293 cells (Life Technologies Cat. No. 11625019) at a ratio of 1:2 and placed in a CO 2 incubator for incubation 4- 5 days. After the expressed antibody was recovered by centrifugation, antibody purification was carried out in accordance with the method of Example 4 to obtain humanized antibody proteins hu2F7 and hu2F8 of the present invention.
  • the cultured MX-1 cells were collected, adjusted for cell density with PBS of pH 7.4, and then plated on a 96-well V-shaped bottom plate with 1 x 105 cells per well. Centrifuge at 2000 rpm for 5 minutes, remove the supernatant, add 100 ⁇ l of the serially diluted chimeric antibody solution (diluted with 0.5% BSA in PBS, 1 ⁇ M starting, 3 fold gradient, 10 doses) per well, mix, shake at 4 °C The bed was incubated for 1 hour; centrifuged at 2000 rpm for 5 minutes, the supernatant was removed, and the cells were washed twice with PBS, and 100 ⁇ l of FITC-labeled goat anti-human secondary antibody (Abeam, cat#ab97224) diluted with 0.5% BSA in PBS was added to each well.
  • FITC-labeled goat anti-human secondary antibody Abeam, cat#ab97224
  • the method steps were the same as the implementation of the 3(1) in vitro indirect ELISA binding assay.
  • the B7-H4 of SEQ ID NO: 92 was decomposed into an antigen fragment of 20 amino acids in length, and the antigen fragment P12 (amino acid sequence: TVASAGNIGEDGILSCTFEP) was found to be specific to the humanized anti-B7-H4 antibody hu2F7 by indirect indirect ELISA. Combined, the results are shown in Figure 2.
  • an alanine scan was performed in the antigen fragment P12, that is, a single amino acid in P12 was separately mutated to alanine.
  • TVASAGNIGEDGILSCTFEP 101 117.9
  • TVASAGNIGEDGALSCTFEP 102 770.6
  • TVASAGNIGEDGIASCTFEP 103 1476
  • TVASAGNIGEDGILSATFEP 104 2006
  • TVASAGNIGEDGILSCTAEP 106 986.5 TVASAGNIGEDGILSCTFAP 107 662.5
  • the antigen fragment P12 (amino acid sequence: TVASAGNIGEDGILSCTFEP) was specifically bound to the humanized anti-B7-H4 antibody hu1C9 by an indirect indirect ELISA binding assay, and the results are shown in the following table.
  • an alanine scan was performed in the antigen fragment P12, that is, a single amino acid in P12 was separately mutated to alanine.
  • Antigenic fragment sequence SEQ ID NO: Affinity EC50(nM) TVASAGNIGEDGILSCTFEP 101 271.7 TVASAGNIGEDGIASCTFEP 103 466.9 TVASAGNIGEDGILSATFEP 104 292.3 TVASAGNIGEDGILSCTAEP 106 487.9
  • the antigen fragment P12 (amino acid sequence: TVASAGNIGEDGILSCTFEP) was specifically bound to the humanized anti-B7-H4 antibody hu2G6 by an indirect indirect ELISA binding assay, and the results are shown in the following table.
  • an alanine scan was performed in the antigen fragment P12, that is, a single amino acid in P12 was separately mutated to alanine.
  • Antigenic fragment sequence SEQ ID NO: Affinity EC50(nM) TVASAGNIGEDGILSCTFEP 101 792.6 TVASAGNIGEDGALSCTFEP 102 2626 TVASAGNIGEDGIASCTFEP 103 3158 TVASAGNIGEDGILSATFEP 104 3480 TVASAGNIGEDGILSCAFEP 105 2289 TVASAGNIGEDGILSCTAEP 106 2600 TVASAGNIGEDGILSCTFEA 108 2372
  • MC38 tumor cells were implanted in mice into which human B7-H4 gene was introduced by gene editing technology, and after the tumor grew to an average of 100 mm 3 , the mice were injected with control or humanized B7-H4 antibody hu1C9 every 3 days ( 10 mg/kg or 30 mg/kg). By observing the size of the tumor, it was found that hu1C9 has a remarkable function of inhibiting tumor growth and has an anti-tumor effect. The specific results are shown in Figure 3 and the following table:
  • Example 10 Effect of humanized antibody on enhancing immunity
  • the anti-tumor effect of anti-B7-H4 may be mediated by enhancing tumor immune effects.
  • different concentrations of anti-B7-H4 antibodies including the humanized antibody hu2G6 and the humanized antibody hu1C9, were added to the cultured T cells.
  • the humanized antibody hu2G6 and the humanized antibody hu1C9 have an effect of increasing T cell proliferation, and can achieve an antitumor effect by enhancing tumor immunity.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • General Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Food Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Cell Biology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
PCT/CN2019/074397 2018-02-11 2019-02-01 抗b7-h4抗体、其抗原结合片段及其医药用途 Ceased WO2019154315A1 (zh)

Priority Applications (13)

Application Number Priority Date Filing Date Title
CN202310820681.5A CN116693686A (zh) 2018-02-11 2019-02-01 抗b7-h4抗体、其抗原结合片段及其医药用途
BR112020015736-8A BR112020015736A2 (pt) 2018-02-11 2019-02-01 Anticorpo anti-b7-h4, fragmento de ligação a antígeno do mesmo e uso farmacêutico do mesmo
RU2020124155A RU2792748C2 (ru) 2018-02-11 2019-02-01 Антитело к b7-h4, его антигенсвязывающий фрагмент и его фармацевтическое применение
CN201980001361.9A CN110366560B (zh) 2018-02-11 2019-02-01 抗b7-h4抗体、其抗原结合片段及其医药用途
KR1020207026012A KR102952441B1 (ko) 2018-02-11 2019-02-01 항 비7 에이치4 항체, 그의 항원 결합 단편 및 그의 약학적 용도
EP19750949.0A EP3753951A4 (en) 2018-02-11 2019-02-01 ANTI-B7-H4 ANTIBODIES, ANTIGEN-BINDING FRAGMENT THEREOF AND PHARMACEUTICAL USE THEREOF
JP2020542138A JP7393337B2 (ja) 2018-02-11 2019-02-01 抗ｂ７－ｈ４抗体、その抗原結合断片及びその医薬用途
CA3089246A CA3089246A1 (en) 2018-02-11 2019-02-01 Anti-b7-h4 antibody, antigen-binding fragment thereof and pharmaceutical use thereof
AU2019218319A AU2019218319B2 (en) 2018-02-11 2019-02-01 Anti-B7-H4 antibody, antigen-binding fragment thereof and pharmaceutical use thereof
US16/967,016 US11472882B2 (en) 2018-02-11 2019-02-01 Anti-B7-H4 antibody, antigen-binding fragment thereof and pharmaceutical use thereof
MX2020008181A MX2020008181A (es) 2018-02-11 2019-02-01 Anticuerpo anti-b7-h4, fragmento de union al antigeno y uso farmaceutico del mismo.
ZA2020/04701A ZA202004701B (en) 2018-02-11 2020-07-29 Anti-b7-h4 antibody, antigen-binding fragment thereof and pharmaceutical use thereof
JP2023197481A JP2024026132A (ja) 2018-02-11 2023-11-21 抗ｂ７－ｈ４抗体、その抗原結合断片及びその医薬用途

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810142118 2018-02-11
CN201810142118.6 2018-02-11

Publications (1)

Publication Number Publication Date
WO2019154315A1 true WO2019154315A1 (zh) 2019-08-15

Family

ID=67548182

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/074397 Ceased WO2019154315A1 (zh) 2018-02-11 2019-02-01 抗b7-h4抗体、其抗原结合片段及其医药用途

Country Status (11)

Country Link
US (1) US11472882B2 (https=)
EP (1) EP3753951A4 (https=)
JP (2) JP7393337B2 (https=)
CN (2) CN116693686A (https=)
AU (1) AU2019218319B2 (https=)
BR (1) BR112020015736A2 (https=)
CA (1) CA3089246A1 (https=)
MX (1) MX2020008181A (https=)
TW (1) TWI823895B (https=)
WO (1) WO2019154315A1 (https=)
ZA (1) ZA202004701B (https=)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020244657A1 (zh) * 2019-06-06 2020-12-10 上海翰森生物医药科技有限公司 抗b7-h4抗体-药物偶联物及其医药用途
WO2022200525A1 (en) 2021-03-26 2022-09-29 Innate Pharma Multispecific proteins comprising an nkp46-binding site, a cancer antgienge binding site fused to a cytokine for nk cell engaging
WO2022258662A1 (en) 2021-06-09 2022-12-15 Innate Pharma Multispecific proteins binding to nkp46, a cytokine receptor, a tumour antigen and cd16a
WO2022258691A1 (en) 2021-06-09 2022-12-15 Innate Pharma Multispecific proteins binding to nkg2d, a cytokine receptor, a tumour antigen and cd16a
WO2022258678A1 (en) 2021-06-09 2022-12-15 Innate Pharma Multispecific proteins binding to nkp30, a cytokine receptor, a tumour antigen and cd16a
US11964024B2 (en) 2021-01-04 2024-04-23 Mersana Therapeutics, Inc. B7H4-targeted antibody-drug conjugates and methods of use thereof
WO2024208818A1 (en) 2023-04-04 2024-10-10 Innate Pharma Modular chimeric antigen receptor
WO2024213081A1 (zh) 2023-04-12 2024-10-17 上海翰森生物医药科技有限公司 抗体药物偶联物在制备预防和/或治疗癌症的药物中的用途

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3753951A4 (en) * 2018-02-11 2022-03-16 Jiangsu Hansoh Pharmaceutical Group Co., Ltd. ANTI-B7-H4 ANTIBODIES, ANTIGEN-BINDING FRAGMENT THEREOF AND PHARMACEUTICAL USE THEREOF
CN115298220B (zh) * 2020-03-24 2024-12-20 上海翰森生物医药科技有限公司 抗体-药物偶联物及其医药用途
TW202144425A (zh) * 2020-04-17 2021-12-01 大陸商上海翰森生物醫藥科技有限公司 特異性抗原結合分子，其製備方法及醫藥用途
CA3183994A1 (en) 2020-06-24 2021-12-30 Philip James Dolan Iii Antibodies recognizing sortilin
IL310780A (en) * 2020-08-18 2024-04-01 Abl Bio Inc Anti-b7-h4/anti-4-1bb bispecific antibodies and use thereof
US20240309095A1 (en) * 2021-07-07 2024-09-19 Incyte Corporation Anti-b7-h4 antibodies and uses thereof
AR127806A1 (es) * 2021-12-01 2024-02-28 Kadmon Corp Llc Anticuerpos b7-h4 y proteínas de fusión de anticuerpo b7-h4 / il-15
JP2025500457A (ja) * 2021-12-23 2025-01-09 プロシーナ バイオサイエンシーズ リミテッド ソルチリンを認識する抗体
CA3245849A1 (en) * 2022-03-30 2025-02-03 Duality Biologics (Suzhou) Co., Ltd. ANTIBODY-DRUG CONJUGATE B7H4 AND ITS USE

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009073533A2 (en) * 2007-11-30 2009-06-11 Medarex, Inc. Anti-b7h4 monoclonal antibody-drug conjugate and methods of use
WO2013025779A1 (en) 2011-08-15 2013-02-21 Amplimmune, Inc. Anti-b7-h4 antibodies and their uses
CN103981150A (zh) * 2014-03-28 2014-08-13 苏州大学 抗人b7-h4单克隆抗体及其制备和应用
US20140322129A1 (en) 2013-03-14 2014-10-30 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
CN104955475A (zh) * 2012-12-19 2015-09-30 艾普利穆恩公司 抗人b7-h4抗体及其用途
CN105189552A (zh) * 2013-03-14 2015-12-23 基因泰克公司 抗b7-h4抗体和免疫缀合物
WO2016040724A1 (en) * 2014-09-12 2016-03-17 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
CN107299085A (zh) * 2017-05-26 2017-10-27 广东医科大学 分泌抗人b7‑h4胞外单克隆抗体杂交瘤细胞株和抗人b7‑h4单克隆抗体及其应用
CN107531782A (zh) * 2015-03-27 2018-01-02 南加利福尼亚大学 用于表达b7‑h4的实体肿瘤的治疗的car t‑细胞

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030091580A1 (en) * 2001-06-18 2003-05-15 Mitcham Jennifer L. Compositions and methods for the therapy and diagnosis of ovarian cancer
US6858710B2 (en) * 1998-12-17 2005-02-22 Corixa Corporation Compositions and methods for the therapy and diagnosis of ovarian cancer
EP3058955B1 (en) * 2005-03-24 2019-05-29 Millennium Pharmaceuticals, Inc. Antibodies that bind ov064 and methods of use therefor
US8759490B2 (en) * 2005-03-24 2014-06-24 Millennium Pharamaceuticals, Inc. Antibodies that bind OV064 and methods of use therefor
CN102875681A (zh) * 2005-07-08 2013-01-16 拜奥根Idec马萨诸塞公司 抗-αvβ6抗体及其用途
PT3042917T (pt) * 2010-08-12 2018-05-02 Lilly Co Eli Anticorpos do péptido beta amilóide anti-n3pglu e seus usos
WO2013157105A1 (ja) * 2012-04-18 2013-10-24 公立大学法人大阪市立大学 ムチンサブタイプ５ａｃ特異的ヒト化抗体およびその利用
WO2018021301A1 (ja) * 2016-07-26 2018-02-01 静岡県 抗ｂ７－ｈ４抗体
EP3753951A4 (en) * 2018-02-11 2022-03-16 Jiangsu Hansoh Pharmaceutical Group Co., Ltd. ANTI-B7-H4 ANTIBODIES, ANTIGEN-BINDING FRAGMENT THEREOF AND PHARMACEUTICAL USE THEREOF
MA52416A (fr) * 2018-03-02 2021-04-21 Five Prime Therapeutics Inc Anticorps b7-h4 et leurs procédés d'utilisation
KR102227515B1 (ko) 2018-12-18 2021-03-12 주식회사 포스코 제올라이트 및 그 제조방법

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009073533A2 (en) * 2007-11-30 2009-06-11 Medarex, Inc. Anti-b7h4 monoclonal antibody-drug conjugate and methods of use
WO2013025779A1 (en) 2011-08-15 2013-02-21 Amplimmune, Inc. Anti-b7-h4 antibodies and their uses
CN104955475A (zh) * 2012-12-19 2015-09-30 艾普利穆恩公司 抗人b7-h4抗体及其用途
US20140322129A1 (en) 2013-03-14 2014-10-30 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
CN105189552A (zh) * 2013-03-14 2015-12-23 基因泰克公司 抗b7-h4抗体和免疫缀合物
CN103981150A (zh) * 2014-03-28 2014-08-13 苏州大学 抗人b7-h4单克隆抗体及其制备和应用
WO2016040724A1 (en) * 2014-09-12 2016-03-17 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
CN107531782A (zh) * 2015-03-27 2018-01-02 南加利福尼亚大学 用于表达b7‑h4的实体肿瘤的治疗的car t‑细胞
CN107299085A (zh) * 2017-05-26 2017-10-27 广东医科大学 分泌抗人b7‑h4胞外单克隆抗体杂交瘤细胞株和抗人b7‑h4单克隆抗体及其应用

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
CHOI IH ET AL., J IMMUNOL., vol. 171, no. 9, 1 November 2003 (2003-11-01), pages 4650 - 4
HOLLIGERHUDSON, NAT. BIOTECHNOL., vol. 23, 2005, pages 1126 - 1136
J. BIOL. CHEM, vol. 243, 1968, pages 3558
KRYCZEK, I. ET AL., J. EXP. MED., vol. 203, no. 4, 2006, pages 871 - 881
NING ZHANG , PENG FANG , ZONG-JIANG GU: "Preparation and Characterization of Monoclonal Antibody Against Human B7-H4 Molecule", MONOCLONAL ANTIBODIES IN IMMUNODIAGNOSIS AND IMMUNOTHERAPY, vol. 33, no. 4, 29 August 2014 (2014-08-29), pages 270 - 274, XP055468610, ISSN: 2167-9436, DOI: 10.1089/mab.2013.0082 *
SICA GL ET AL., IMMUNITY, vol. 18, no. 6, June 2003 (2003-06-01), pages 849 - 61
SUH WK ET AL., BLOOD. MOL CELL BIOL., vol. 26, no. 17, September 2006 (2006-09-01), pages 6403 - 11
ZHANG BIAO , FANG PENG , ZHANG NING , GU ZONG-JIANG : "The Preparation of Anti-Human B7-H4 Monoclonal Antibody and Its Function Study", CHINESE JOURNAL OF HEMORHEOLOGY, vol. 25, no. 1, 31 December 2015 (2015-12-31), pages 18 - 21, XP009523002, ISSN: 1009-881x, DOI: 10.3969./j.issn.1009-881x.2015.01.005 *
ZHU G ET AL., BLOOD, vol. 113, no. 8, 19 February 2009 (2009-02-19), pages 1759 - 67

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020244657A1 (zh) * 2019-06-06 2020-12-10 上海翰森生物医药科技有限公司 抗b7-h4抗体-药物偶联物及其医药用途
CN112351797A (zh) * 2019-06-06 2021-02-09 上海翰森生物医药科技有限公司 抗b7-h4抗体-药物偶联物及其医药用途
US20230072897A1 (en) * 2019-06-06 2023-03-09 Shanghai Hansoh Biomedical Co., Ltd. Anti-b7-h4 antibody-drug conjugate and medicinal use thereof
CN112351797B (zh) * 2019-06-06 2023-12-22 上海翰森生物医药科技有限公司 抗b7-h4抗体-药物偶联物及其医药用途
US11964024B2 (en) 2021-01-04 2024-04-23 Mersana Therapeutics, Inc. B7H4-targeted antibody-drug conjugates and methods of use thereof
WO2022200525A1 (en) 2021-03-26 2022-09-29 Innate Pharma Multispecific proteins comprising an nkp46-binding site, a cancer antgienge binding site fused to a cytokine for nk cell engaging
WO2022258662A1 (en) 2021-06-09 2022-12-15 Innate Pharma Multispecific proteins binding to nkp46, a cytokine receptor, a tumour antigen and cd16a
WO2022258691A1 (en) 2021-06-09 2022-12-15 Innate Pharma Multispecific proteins binding to nkg2d, a cytokine receptor, a tumour antigen and cd16a
WO2022258678A1 (en) 2021-06-09 2022-12-15 Innate Pharma Multispecific proteins binding to nkp30, a cytokine receptor, a tumour antigen and cd16a
WO2024208818A1 (en) 2023-04-04 2024-10-10 Innate Pharma Modular chimeric antigen receptor
WO2024213081A1 (zh) 2023-04-12 2024-10-17 上海翰森生物医药科技有限公司 抗体药物偶联物在制备预防和/或治疗癌症的药物中的用途
EP4696330A1 (en) 2023-04-12 2026-02-18 Shanghai Hansoh Biomedical Co., Ltd. Use of antibody-drug conjugate in preparation of drug for preventing and/or treating cancer

Also Published As

Publication number Publication date
BR112020015736A2 (pt) 2020-12-08
AU2019218319B2 (en) 2025-11-13
JP2024026132A (ja) 2024-02-28
US11472882B2 (en) 2022-10-18
US20210032347A1 (en) 2021-02-04
TWI823895B (zh) 2023-12-01
ZA202004701B (en) 2023-12-20
AU2019218319A1 (en) 2020-08-13
CN116693686A (zh) 2023-09-05
TW201934581A (zh) 2019-09-01
KR20200119846A (ko) 2020-10-20
JP7393337B2 (ja) 2023-12-06
EP3753951A1 (en) 2020-12-23
CN110366560B (zh) 2023-07-28
CA3089246A1 (en) 2019-08-15
JP2021513331A (ja) 2021-05-27
MX2020008181A (es) 2020-09-18
CN110366560A (zh) 2019-10-22
EP3753951A4 (en) 2022-03-16
RU2020124155A (ru) 2022-03-11

Similar Documents

Publication Publication Date Title
US11472882B2 (en) Anti-B7-H4 antibody, antigen-binding fragment thereof and pharmaceutical use thereof
TWI673287B (zh) 抗b7-h3抗體、其抗原結合片段及其醫藥用途
TWI840399B (zh) 結合人il-4r的抗體、其抗原結合片段及其醫藥用途
US11525005B2 (en) Anti-CD40 antibody, antigen binding fragment thereof and medical use thereof
WO2019141268A1 (zh) 抗4-1bb抗体、其抗原结合片段及其医药用途
CN112243443B (zh) 抗trop-2抗体、其抗原结合片段及其医药用途
CN110461874B (zh) 抗gitr抗体、其抗原结合片段及其医药用途
CN113227148A (zh) 抗gpc3抗体、其抗原结合片段及其医药用途
CN115298216A (zh) 抗体或其抗原结合片段、其制备方法及医药用途
WO2019080889A1 (zh) 抗csf-1r抗体、其抗原结合片段及其医药用途
CN115109157A (zh) 抗体或其抗原结合片段，其制备方法及医药用途
WO2021209066A1 (zh) 特异性抗原结合分子，其制备方法及医药用途
WO2022078490A1 (zh) 抗erbb3抗体或其抗原结合片段及其医药用途
KR102952441B1 (ko) 항 비7 에이치4 항체, 그의 항원 결합 단편 및 그의 약학적 용도
RU2792748C2 (ru) Антитело к b7-h4, его антигенсвязывающий фрагмент и его фармацевтическое применение
HK40012331B (zh) 抗b7-h4抗体、其抗原结合片段及其医药用途
KR20260053095A (ko) 항 비7 에이치4 항체, 그의 항원 결합 단편 및 그의 약학적 용도
HK40036076A (en) Anti-trop-2 antibody, antigen binding fragment thereof and medical use thereof
HK40036076B (en) Anti-trop-2 antibody, antigen binding fragment thereof and medical use thereof
HK40012331A (en) Anti-b7-h4 antibody, antigen-binding fragment thereof and pharmaceutical use thereof

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: 19750949

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 3089246

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2020542138

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2019218319

Country of ref document: AU

Date of ref document: 20190201

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 20207026012

Country of ref document: KR

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 2019750949

Country of ref document: EP

Effective date: 20200911

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112020015736

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 112020015736

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20200731