WO2022188652A1 - Ror1结合蛋白及其用途 - Google Patents

Ror1结合蛋白及其用途 Download PDF

Info

Publication number
WO2022188652A1
WO2022188652A1 PCT/CN2022/078327 CN2022078327W WO2022188652A1 WO 2022188652 A1 WO2022188652 A1 WO 2022188652A1 CN 2022078327 W CN2022078327 W CN 2022078327W WO 2022188652 A1 WO2022188652 A1 WO 2022188652A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
sequence
cdr
heavy chain
light chain
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/CN2022/078327
Other languages
English (en)
French (fr)
Chinese (zh)
Inventor
田海军
陈扬德
张盈华
朱志强
吴文慧
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.)
Sichuan Kelun Biotech Biopharmaceutical Co Ltd
Klus Pharma Inc
Original Assignee
Sichuan Kelun Biotech Biopharmaceutical Co Ltd
Klus Pharma Inc
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 Sichuan Kelun Biotech Biopharmaceutical Co Ltd, Klus Pharma Inc filed Critical Sichuan Kelun Biotech Biopharmaceutical Co Ltd
Priority to CA3207635A priority Critical patent/CA3207635A1/en
Priority to JP2023541591A priority patent/JP2024508597A/ja
Priority to US18/260,556 priority patent/US20240018237A1/en
Priority to EP22766178.2A priority patent/EP4306540A4/en
Priority to CN202280009364.9A priority patent/CN116829599A/zh
Priority to MX2023008176A priority patent/MX2023008176A/es
Priority to KR1020237024374A priority patent/KR20230156021A/ko
Publication of WO2022188652A1 publication Critical patent/WO2022188652A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
    • 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
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • 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
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • 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
    • C07K2317/56—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
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • 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
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/734—Complement-dependent cytotoxicity [CDC]
    • 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
    • C07K2317/77—Internalization into the cell
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]

Definitions

  • the present invention belongs to the field of therapeutic monoclonal antibodies, more particularly, the present invention relates to an antibody against ROR1; and also relates to the use of the antibody in the treatment of diseases.
  • Receptor tyrosine kinases are multigroup transmembrane proteins that act on receptors for cytokines, growth factors, hormones and other signaling molecules and play important roles in a variety of cellular processes, including Growth, differentiation, angiogenesis, and progression of various cancers.
  • RTKs Receptor tyrosine kinase-like orphan receptor 1
  • ROR1 Receptor tyrosine kinase-like orphan receptor 1
  • ROR1 is a novel drug target with broad-spectrum anticancer potential.
  • scFvs single-chain fragment variables
  • chimeric antibodies chimeric antibodies
  • Fab antibodies antibody drug conjugates
  • ADC antibody-drug conjugate
  • CAR chimeric antigen receptor
  • BiAb bispecific antibody
  • the anti-ROR1 monoclonal antibody with rapid research progress is cirmtuzumab from Oncternal Therapeutics. Its clinical data has a good anti-cancer effect, which can better inhibit the proliferation, migration and survival of chronic leukemia tumor cells.
  • the inventors first developed a murine antibody with excellent properties, which can specifically recognize/bind ROR1, but not ROR2. On this basis, the inventors have devoted a lot of creative work, conducted in-depth research and transformation of the mouse-derived antibody, and developed the chimeric antibody and humanized antibody of the mouse-derived antibody.
  • the humanized antibody of the present invention not only has a very high degree of humanization, but also has the same heavy chain variable region and light chain variable region as the murine antibody and human-mouse chimeric antibody (chimeric antibody has exactly the same heavy chain variable region and light chain as the mouse antibody). variable region) have substantially the same (or even better) biological function.
  • the antibodies of the present invention are extremely advantageous, which not only retain the functions and properties of the parental murine antibody, such as binding to ROR1 with high affinity and specificity, but thus have the potential to prevent and the potential to treat tumors; in addition, the antibodies of the present invention can more effectively induce ROR1-mediated endocytosis when they bind to ROR1 on the cell surface, thereby serving as carriers for targeted therapy.
  • the humanized antibodies of the present invention have an extremely high degree of humanization and thus can be safely administered to human subjects without eliciting an immunogenic response. Therefore, the antibody of the present invention has great clinical value.
  • the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to ROR1.
  • the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding the extracellular domain (ECD) of ROR1.
  • ECD extracellular domain
  • CDRs complementarity determining regions
  • CDR-L1, CDR-L2 and CDR-L3 contained in the light chain variable region (VL) shown in any of SEQ ID NO: 2 and 107-110;
  • CDR-L1, CDR-L2 and CDR-L3 contained in the light chain variable region (VL) shown in any of SEQ ID NO: 4 and 87-101;
  • CDR-L1, CDR-L2 and CDR-L3 contained in the light chain variable region (VL) shown in any of SEQ ID NOs: 6, 107, 116-118;
  • At least one CDR contains a mutation that is a substitution, deletion or addition of one or several amino acids (eg, a substitution, deletion or addition of 1, 2 or 3 amino acids); preferably , the substitutions are conservative substitutions.
  • the CDRs are defined according to the Chothia, AbM, Kabat or IMGT numbering systems; preferably, the VH and/or VL of the antibody or antigen-binding fragment thereof includes an immunoglobulin from human or murine The framework regions (FRs) of the protein, preferably, the antibody or antigen-binding fragment thereof binds human ROR1.
  • the VH and/or VL of the antibody or antigen-binding fragment thereof includes an immunoglobulin from human or murine The framework regions (FRs) of the protein, preferably, the antibody or antigen-binding fragment thereof binds human ROR1.
  • an antibody or antigen-binding fragment thereof capable of specifically binding to ROR1 comprising: a variable heavy chain (VH) and/or a variable light chain ( VL).
  • VH variable heavy chain
  • VL variable light chain
  • the antibodies or antigen-binding fragments thereof of the invention comprise the following heavy chain variable regions (VH) and/or light chain variable regions (VL), wherein the CDRs are defined according to the Chothia numbering system:
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 shown in any of SEQ ID NOs: 48-55, and the sequence is CDR-L2 shown in any of SEQ ID NOs: 56-58 , the sequence is the CDR-L3 shown in SEQ ID NO: 44;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 shown in any of SEQ ID NOs: 21-23, 77, and the sequence is CDR-L2 shown in SEQ ID NO: 24 or 25 , the sequence is the CDR-L3 shown in SEQ ID NO: 15;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is the CDR-L1 shown in any of SEQ ID NOs: 75-77, the sequence is the CDR-L2 shown in SEQ ID NO: 24 or 78, the sequence It is CDR-L3 shown in SEQ ID NO: 15.
  • the antibody or antigen-binding fragment thereof comprises the following CDRs as defined by the Chothia numbering system:
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:48, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:48, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:49, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:50, the sequence is CDR-L2 of SEQ ID NO:58, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:51, CDR-L2 of sequence SEQ ID NO:56, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:52, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:53, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:54, CDR-L2 of sequence SEQ ID NO:56, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:55, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:21, CDR-L2 of sequence SEQ ID NO:24, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:75, CDR-L2 of sequence SEQ ID NO:24, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:76, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 .
  • the antibodies or antigen-binding fragments thereof of the invention comprise the following heavy chain variable regions (VH) and/or light chain variable regions (VL), wherein the CDRs are defined by the AbM numbering system:
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 shown in any of SEQ ID NOs: 48-55, and the sequence is CDR-L2 shown in any of SEQ ID NOs: 56-58 , the sequence is the CDR-L3 shown in SEQ ID NO: 44;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 shown in any of SEQ ID NOs: 21-23, 77, and the sequence is CDR-L2 shown in SEQ ID NO: 24 or 25 , the sequence is the CDR-L3 shown in SEQ ID NO: 15;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is the CDR-L1 shown in any of SEQ ID NO: 75-77, the sequence is the CDR-L2 shown in SEQ ID NO: 24 or 78, the sequence It is CDR-L3 shown in SEQ ID NO: 15.
  • the antibody or antigen-binding fragment thereof comprises the following CDRs as defined by the AbM numbering system:
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:48, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:48, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:49, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • a light chain variable region comprising the following 3 CDRs: CDR-L1 of SEQ ID NO:50, CDR-L2 of SEQ ID NO:58, CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:51, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:52, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:53, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:54, CDR-L2 of sequence SEQ ID NO:56, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:55, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:21, CDR-L2 of sequence SEQ ID NO:24, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:75, CDR-L2 of sequence SEQ ID NO:24, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:76, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 .
  • the antibodies or antigen-binding fragments thereof of the invention comprise the following heavy chain variable regions (VH) and/or light chain variable regions (VL), wherein the CDRs are defined according to the Kabat numbering system:
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 shown in any of SEQ ID NOs: 48-55, and the sequence is CDR-L2 shown in any of SEQ ID NOs: 56-58 , the sequence is the CDR-L3 shown in SEQ ID NO: 44;
  • VH heavy chain variable region
  • the light chain variable region comprising the following 3 CDRs: the sequence is the CDR-L1 shown in any of SEQ ID NOs: 21-23, 77, and the sequence is the CDR shown in any of SEQ ID NO: 24-25 -L2, the sequence is CDR-L3 shown in SEQ ID NO: 15;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is the CDR-L1 shown in any of SEQ ID NOs: 75-77, the sequence is the CDR-L2 shown in SEQ ID NO: 24 or 78, the sequence It is CDR-L3 shown in SEQ ID NO: 15.
  • the antibody or antigen-binding fragment thereof comprises the following CDRs as defined by the Kabat numbering system:
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:48, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:48, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:48, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:49, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:50, the sequence is CDR-L2 of SEQ ID NO:58, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:51, CDR-L2 of sequence SEQ ID NO:56, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:48, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:48, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:49, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:50, the sequence is CDR-L2 of SEQ ID NO:58, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:51, CDR-L2 of sequence SEQ ID NO:56, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:48, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:49, CDR-L2 of sequence SEQ ID NO:57, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:50, the sequence is CDR-L2 of SEQ ID NO:58, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:51, CDR-L2 of sequence SEQ ID NO:56, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:52, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:53, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:54, CDR-L2 of sequence SEQ ID NO:56, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:55, the sequence is CDR-L2 of SEQ ID NO:56, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:21, CDR-L2 of sequence SEQ ID NO:24, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:22, the sequence is CDR-L2 of SEQ ID NO:25, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:23, CDR-L2 of sequence SEQ ID NO:25, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:75, CDR-L2 of sequence SEQ ID NO:24, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:76, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:77, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:76, CDR-L2 of sequence SEQ ID NO:78, CDR-L3 of sequence SEQ ID NO:15 .
  • the antibodies or antigen-binding fragments thereof of the invention comprise the following heavy chain variable regions (VH) and/or light chain variable regions (VL), wherein the CDRs are defined by the IMGT numbering system:
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: sequence is CDR-L1 shown in any of SEQ ID NO:38-42, sequence is CDR-L2 shown in SEQ ID NO:43, sequence is SEQ ID NO:43 CDR-L3 shown in ID NO: 44;
  • VH heavy chain variable region
  • the light chain variable region comprising the following 3 CDRs: sequence is CDR-L1 shown in SEQ ID NO: 12 or 13, sequence is CDR-L2 shown in SEQ ID NO: 14, sequence is SEQ ID NO : CDR-L3 shown in 15;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: sequence is CDR-L1 shown in SEQ ID NO: 69 or 70, sequence is CDR-L2 shown in SEQ ID NO: 71, sequence is SEQ ID NO : CDR-L3 shown in 15.
  • the antibody or antigen-binding fragment thereof comprises the following CDRs as defined by the IMGT numbering system:
  • VH heavy chain variable region comprising the following 3 CDRs: CDR-H1 of sequence SEQ ID NO:35, CDR-H2 of sequence SEQ ID NO:36, sequence of CDR-H2 of SEQ ID NO:37 CDR-H3; and/or,
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:38, the sequence is CDR-L2 of SEQ ID NO:43, the sequence is CDR-L3 of SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:39, CDR-L2 of sequence SEQ ID NO:43, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:40, CDR-L2 of sequence SEQ ID NO:43, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:41, CDR-L2 of sequence SEQ ID NO:43, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • VL The light chain variable region (VL) comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:42, CDR-L2 of sequence SEQ ID NO:43, CDR-L3 of sequence SEQ ID NO:44 ;
  • VH heavy chain variable region
  • a light chain variable region comprising the following 3 CDRs: CDR-L1 of SEQ ID NO: 12, CDR-L2 of SEQ ID NO: 14, CDR-L3 of SEQ ID NO: 15 ;
  • VH heavy chain variable region
  • a light chain variable region comprising the following 3 CDRs: CDR-L1 of SEQ ID NO: 13, CDR-L2 of SEQ ID NO: 14, CDR-L3 of SEQ ID NO: 15 ;
  • VH heavy chain variable region
  • a light chain variable region comprising the following 3 CDRs: CDR-L1 of SEQ ID NO: 12, CDR-L2 of SEQ ID NO: 14, CDR-L3 of SEQ ID NO: 15 ;
  • VH heavy chain variable region
  • a light chain variable region comprising the following 3 CDRs: CDR-L1 of SEQ ID NO: 13, CDR-L2 of SEQ ID NO: 14, CDR-L3 of SEQ ID NO: 15 ;
  • VH heavy chain variable region
  • a light chain variable region comprising the following 3 CDRs: CDR-L1 of SEQ ID NO: 12, CDR-L2 of SEQ ID NO: 14, CDR-L3 of SEQ ID NO: 15 ;
  • VH heavy chain variable region
  • a light chain variable region comprising the following 3 CDRs: CDR-L1 of SEQ ID NO: 13, CDR-L2 of SEQ ID NO: 14, CDR-L3 of SEQ ID NO: 15 ;
  • VH heavy chain variable region
  • the light chain variable region (VL) comprising the following 3 CDRs: the sequence is CDR-L1 of SEQ ID NO:69, the sequence is CDR-L2 of SEQ ID NO:71, the sequence is CDR-L3 of SEQ ID NO:15 ;
  • VH heavy chain variable region
  • the light chain variable region comprising the following 3 CDRs: CDR-L1 of sequence SEQ ID NO:70, CDR-L2 of sequence SEQ ID NO:71, CDR-L3 of sequence SEQ ID NO:15 .
  • the antibodies or antigen-binding fragments thereof of the invention comprise a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein the heavy chain variable region (VH) ) and/or light chain variable region (VL) compared to the heavy chain variable region and/or light chain variable region described in any one of the aforementioned Chothia, AbM, Kabat or IMGT definitions, at least one CDR Mutations are included which are substitutions, deletions or additions of one or several amino acids (eg, substitutions, deletions or additions of 1, 2 or 3 amino acids); preferably, the substitutions are conservative substitutions.
  • the VH of an antibody or antigen-binding fragment thereof of the invention comprises a heavy chain variable region (VH) framework region (FR) derived from a murine immunoglobulin, and/or the antibody or its
  • the VL of the antigen-binding fragment comprises a light chain variable region (VL) framework region (FR) derived from a murine immunoglobulin.
  • the antibodies or antigen-binding fragments thereof of the invention are of murine origin.
  • the VH of an antibody or antigen-binding fragment thereof of the invention comprises a heavy chain variable region (VH) framework region (FR) derived from a human immunoglobulin, and/or the antibody or its
  • the VL of the antigen-binding fragment comprises a light chain variable region (VL) framework region (FR) derived from a human immunoglobulin. Accordingly, in certain preferred embodiments, the antibodies or antigen-binding fragments thereof of the invention are humanized.
  • the heavy chain variable region FR and/or light chain variable region FR of the antibody or antigen-binding fragment thereof of the invention may comprise one or more non-human (eg, murine) amino acid residues
  • the heavy chain framework region FR and/or the light chain framework region FR may comprise one or more amino acid back-mutations in which there are corresponding murine amino acid residues.
  • the antibody or antigen-binding fragment thereof of the invention comprises:
  • a heavy chain framework region of a human immunoglobulin or a variant thereof that has conservative substitutions of up to 20 amino acids (eg, up to 15, up to 10) compared to the germline antibody gene sequence from which it is derived , or conservative substitutions of up to 5 amino acids; e.g., conservative substitutions of 1, 2, 3, 4 or 5 amino acids); and/or
  • a light chain framework region of a human immunoglobulin or a variant thereof that has conservative substitutions of up to 20 amino acids (eg, up to 15, up to 10) compared to the sequence of the germline antibody gene from which it is derived , or conservative substitutions of up to 5 amino acids; eg, conservative substitutions of 1, 2, 3, 4 or 5 amino acids).
  • the antibody or antigen-binding fragment thereof of the invention is at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93% humanized %, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%.
  • the antibody or antigen-binding fragment thereof of the invention comprises:
  • VH heavy chain variable region
  • (iii) have at least 75%, at least 80%, at least 85%, at least 90%, at least Sequences of 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;
  • VL light chain variable region
  • the antibodies or antigen-binding fragments thereof of the invention have the following heavy chain variable regions (VH) and/or light chain variable regions (VL): SEQ ID NOs: 1, 3, 5, 84 - the VH sequence shown in any one of 86, 102-106, 111-115, and/or, the VL shown in any one of SEQ ID NOs: 2, 4, 6, 87-101, 107-110, 116-118 the sequence of.
  • VH heavy chain variable regions
  • VL light chain variable regions
  • the antibodies or antigen-binding fragments thereof of the invention have the following heavy chain variable regions (VH) and/or light chain variable regions (VL): any of SEQ ID NOs: 86, 102, 114
  • VH heavy chain variable regions
  • VL light chain variable regions
  • the VH and/or VL of the antibody or antigen-binding fragment thereof has the VH sequence set forth in any one of SEQ ID NOs: 86, 102, 114 and/or SEQ ID NOs: 92, 107 , at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96% compared to the VL sequences shown in any one of 117 %, at least 97%, at least 98%, at least 99%, or 100% sequence identity.
  • the VH and/or VL of the antibody or antigen-binding fragment thereof is the same as the VH sequence set forth in any one of SEQ ID NOs: 86, 102, 114 and/or SEQ ID NOs: 92, 107, 117 has one or more amino acid substitutions, deletions or additions (eg, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) compared to the VL sequences shown in any one of 117.
  • the substitutions are conservative substitutions.
  • the antibody or antigen-binding fragment thereof of the invention comprises:
  • the heavy chain variable region (VH) and light chain variable region (VL) are compared to any one of groups (1) to (68), the heavy chain variable region (VH) ) has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity; and/or, the light chain variable region (VL) has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity.
  • the heavy chain variable region (VH) and light chain variable region (VL) are compared to any one of groups (1) to (68), the heavy chain variable region (VH) ) with one or several amino acid substitutions, deletions or additions (e.g. 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions); and/or, the light chain variable region (VL) There are substitutions, deletions or additions of one or several amino acids (eg substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids); preferably, the substitutions are conservative substitutions.
  • the antibodies or antigen-binding fragments thereof of the invention may further comprise constant region sequences derived from mammalian (eg, murine or human) immunoglobulins or variants thereof.
  • the heavy chain of an antibody or antigen-binding fragment thereof of the invention comprises a heavy chain constant region (CH) of a human or murine immunoglobulin or a variant thereof that is the same as the wild-type from which it was derived
  • the sequence has one or more amino acid substitutions, deletions, or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions; e.g., 1, 2, 3 and/or
  • the light chain of the antibody or antigen-binding fragment thereof of the invention comprises the light chain constant region (CL) of a human or murine immunoglobulin or a variant thereof
  • a variant that has one or more amino acid substitutions, deletions, or additions e.g., substitutions of up to 20, up to 15, up to
  • the heavy chain of an antibody or antigen-binding fragment thereof of the invention comprises the heavy chain constant region (CH) of a human immunoglobulin or a variant thereof, the variant and the wild-type sequence from which it is derived compared to conservative substitutions of up to 20 amino acids (e.g., conservative substitutions of up to 15, up to 10, or up to 5 amino acids; e.g., conservative substitutions of 1, 2, 3, 4, or 5 amino acids);
  • the light chain of the antibody or antigen-binding fragment thereof of the invention comprises the light chain constant region (CL) of a human immunoglobulin or a variant thereof having at most Conservative substitutions of 20 amino acids (eg, conservative substitutions of up to 15, up to 10, or up to 5 amino acids; eg, conservative substitutions of 1, 2, 3, 4, or 5 amino acids).
  • the heavy chain of an antibody or antigen-binding fragment thereof of the invention comprises the heavy chain constant region (CH) of a murine immunoglobulin or a variant thereof that is identical to the wild-type sequence from which it was derived than conservative substitutions of up to 20 amino acids (e.g., conservative substitutions of up to 15, up to 10, or up to 5 amino acids; e.g., conservative substitutions of 1, 2, 3, 4, or 5 amino acids); and /or, the light chain of the antibody or antigen-binding fragment thereof of the invention comprises the light chain constant region (CL) of a murine immunoglobulin or a variant thereof having at most 20 ⁇ compared to the wild-type sequence from which it is derived. Conservative substitutions of amino acids (eg, conservative substitutions of up to 15, up to 10, or up to 5 amino acids; eg, conservative substitutions of 1, 2, 3, 4, or 5 amino acids).
  • CH heavy chain constant region
  • the light chain of the antibody or antigen-binding fragment thereof of the invention comprises the light chain constant region (CL) of
  • the constant region is altered, eg, mutated, to modify properties of the anti-ROR1 antibody molecule (eg, to alter one or more of the following: Fc receptor binding, antibody glycosylation, cysteine number of residues, effector cell function or complement function).
  • Changes in effector function e.g., enhancement of an antibody's affinity for an effector ligand (e.g., FcR or complement C1q) can be produced by substituting at least one amino acid residue in the constant region of an antibody for a different residue, resulting in a change in function, e.g. ).
  • the Fc region of an antibody mediates several important effector functions such as ADCC, phagocytosis, CDC, etc. In certain instances, these effector functions are required for therapeutic antibodies.
  • the anti-ROR1 antibody molecule has a heavy chain constant region (Fc) selected from, for example, the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD and IgE; particularly From, for example, heavy chain constant regions of IgG1, IgG2, IgG3 and IgG4, more particularly selected from heavy chain constant regions of IgG1 or IgG4 (eg human IgG1 or IgG4).
  • Fc heavy chain constant region
  • the anti-ROR1 antibody molecule has a light chain constant region selected from, eg, a kappa or lambda light chain constant region, preferably a kappa light chain constant region (eg, a human kappa light chain).
  • the antibody or antigen-binding fragment thereof comprises a heavy chain constant region selected from the group consisting of:
  • the antibody or antigen-binding fragment thereof of the invention comprises:
  • a heavy chain constant region comprising an amino acid sequence selected from the group consisting of:
  • a light chain constant region comprising an amino acid sequence selected from the group consisting of:
  • substitutions described in (ii) or (v) are conservative substitutions.
  • the antibodies or antigen-binding fragments thereof of the invention comprise a heavy chain constant region (CH) as set forth in SEQ ID NO:119 and a light chain constant region (CL) as set forth in SEQ ID NO:120 ).
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 1 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 2 and the light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 3 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 4 and the light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 5 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 6 and the light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 84 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 87 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 84 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 88 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 84 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 89 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 84 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 90 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 84 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 91 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 85 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 87 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 85 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 88 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 85 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 92 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 85 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 90 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 85 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 91 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 87 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 88 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 89 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 90 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 91 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 92 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 93 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 94 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 95 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 96 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 97 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 98 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 99 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 100 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 86 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 101 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 102 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 102 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 108 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 102 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 109 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 102 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 110 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 103 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 103 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 108 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 103 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 109 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 103 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 110 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 104 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 104 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 108 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 104 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 109 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 104 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 110 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 105 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 105 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 108 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 105 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 109 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 105 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 110 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 106 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 106 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 108 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 106 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 109 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 106 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 110 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 111 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 111 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 116 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 111 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 117 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 111 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 118 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 112 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 112 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 116 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 112 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 117 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 112 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 118 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 113 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 113 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 116 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 113 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 117 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 113 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 118 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 114 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 114 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 116 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 114 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 117 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 114 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 118 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 115 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 107 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 115 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 116 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 115 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 117 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention comprise the VH of the sequence set forth in SEQ ID NO: 115 and the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119, and, comprise the heavy chain of the heavy chain constant region (CH) set forth in SEQ ID NO: 119 : VL of the sequence shown in 118 and light chain of the light chain constant region (CL) shown in SEQ ID NO: 120.
  • the antibodies of the invention are chimeric or humanized antibodies.
  • the antibody or antigen-binding fragment thereof of the invention is selected from the group consisting of ScFv, Fab, Fab', (Fab') 2 , Fv fragment, disulfide-linked Fv (dsFv), diabody ), bispecific antibodies, and multispecific antibodies.
  • the antibody molecules or antigen-binding fragments thereof of the invention may exhibit at least one of the following properties:
  • ROR1 binds ROR1 (eg, human ROR1 ) with a K of less than about 100 nM, eg, less than about 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 1 nM, 0.1 nM, or less; eg, by biofilm interference techniques (BLI) (eg ForteBio ) measured;
  • BLI biofilm interference techniques
  • (b) binds ROR1 with an EC50 of less than about 500 nM, eg, less than about 100 nM, 10 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM or less (eg, human ROR1); eg, the EC50 is determined by flow cytometry or competitive ELISA techniques;
  • the antibody or antigen-binding fragment thereof does not bind ROR2 (eg, human ROR2).
  • the antibody or antigen-binding fragment thereof has ADCC activity.
  • the antibody or antigen-binding fragment thereof has CDC activity.
  • the antibody or antigen-binding fragment thereof has ADCC and CDC activities.
  • the antibody or antigen-binding fragment thereof has enhanced ADCC and/or CDC activity.
  • the antibodies or antigen-binding fragments thereof of the present invention possess at least one of the following biological functions:
  • the antibodies or antigen-binding fragments thereof of the invention possess any combination of the aforementioned biological functions.
  • An antibody or antigen-binding fragment thereof of the invention can be derivatized, eg, linked to another molecule (eg, another polypeptide or protein).
  • another molecule eg, another polypeptide or protein.
  • derivatization eg, labeling
  • the antibodies or antigen-binding fragments thereof of the invention are also intended to include such derivatized forms.
  • an antibody or antigen-binding fragment thereof of the invention can be functionally linked (by chemical coupling, genetic fusion, non-covalent attachment, or otherwise) to one or more other molecular moieties, such as another antibody (eg, to form a bispecific antibodies), detection reagents, pharmaceutical reagents, and/or proteins or polypeptides (eg, avidin or polyhistidine tags) capable of mediating binding of an antibody or antigen-binding fragment to another molecule.
  • another antibody eg, to form a bispecific antibodies
  • detection reagents eg, to form a bispecific antibodies
  • pharmaceutical reagents eg, to form a bispecific antibodies
  • proteins or polypeptides eg, avidin or polyhistidine tags
  • bispecific antibody is produced by cross-linking two or more antibodies (of the same or different types).
  • Methods of obtaining bispecific antibodies are well known in the art, examples of which include, but are not limited to, chemical cross-linking methods, cell engineering methods (hybridoma methods), or genetic engineering methods.
  • Another type of derivatized antibody is a labeled antibody.
  • an antibody or antigen-binding fragment thereof of the invention can be linked to a detectable label.
  • the detectable label of the present invention can be any substance detectable by fluorescent, spectroscopic, photochemical, biochemical, immunological, electrical, optical or chemical means.
  • Such labels are well known in the art, examples of which include, but are not limited to, enzymes (eg, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, urease, glucose oxidase, etc.), radionuclides Fluorescein (eg, 3H, 125I, 35S, 14C, or 32P), fluorescent dyes (eg, fluorescein isothiocyanate (FITC), fluorescein, tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin ( PE), Texas red, rhodamine, quantum dots or cyanine derivatives (eg Cy7, Alexa 750)), acridine esters, magnetic beads (eg, ), calorimetric labels such as colloidal gold or colored glass or plastic (eg, polystyrene, polypropylene, latex, etc.) beads, and modified avidins (eg, streptavidin) for
  • Patents teaching the use of this marker include, but are not limited to, US Patents 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241 (all incorporated herein by reference).
  • Detectable labels as described above can be detected by methods known in the art. For example, radiolabels can be detected using photographic film or a scintillation calculator, and fluorescent labels can be detected using a light detector to detect the emitted light.
  • Enzyme labels are generally detected by providing a substrate to the enzyme and detecting the reaction product produced by the action of the enzyme on the substrate, and calorimetric labels are detected by simply visualizing the colored label.
  • such labels can be suitable for use in immunological detection (eg, enzyme-linked immunoassays, radioimmunoassays, fluorescent immunoassays, chemiluminescence immunoassays, etc.).
  • a detectable label as described above can be attached to an antibody or antigen-binding fragment thereof of the invention via linkers of various lengths to reduce potential steric hindrance.
  • the antibodies or antigen-binding fragments thereof of the present invention can also be derivatized with chemical groups such as polyethylene glycol (PEG), methyl or ethyl, or glycosyl groups. These groups can be used to improve the biological properties of antibodies, such as increasing serum half-life.
  • PEG polyethylene glycol
  • methyl or ethyl methyl or ethyl
  • glycosyl groups can be used to improve the biological properties of antibodies, such as increasing serum half-life.
  • the present invention provides a conjugate comprising the monoclonal antibody of the present invention or an antigen-binding fragment thereof and a coupling moiety selected from the group consisting of detectable labels, radioisotopes , Fluorescent substances, luminescent substances, colored substances, enzymes, polyethylene glycol (PEG), nuclides, nucleic acids, small molecule toxins, with binding active polypeptides, proteins, receptors, ligands, and other inhibiting tumor cell growth, promoting Active substances of tumor cell apoptosis or necrosis.
  • a coupling moiety selected from the group consisting of detectable labels, radioisotopes , Fluorescent substances, luminescent substances, colored substances, enzymes, polyethylene glycol (PEG), nuclides, nucleic acids, small molecule toxins, with binding active polypeptides, proteins, receptors, ligands, and other inhibiting tumor cell growth, promoting Active substances of tumor cell apoptosis or necrosis.
  • the present invention provides a chimeric antigen receptor comprising the monoclonal antibody or an antigen-binding fragment thereof.
  • the chimeric antigen receptor comprises a monoclonal antibody of the invention or an antigen-binding fragment thereof (eg, ScFv), a transmembrane domain, and one or more intracellular T cell signaling structures area.
  • the present invention also provides host cells containing or expressing the chimeric antigen receptor, such as immune cells (eg T lymphocytes, NK cells).
  • the present invention provides a multispecific antibody
  • the antibody is formed by coupling a first antibody or its fragment and other antibodies or its fragments or antibody analogs, each antibody or its fragment or antibody The analog retains the original binding specificity, and the first antibody or its fragment is the antibody or its antigen-binding fragment of the present invention; preferably, the multispecific antibody is a bispecific antibody or a trispecific antibody or a tetraspecific antibody. specific antibodies.
  • the antibodies of the present invention can be prepared by various methods known in the art, such as by genetic engineering recombinant techniques. For example, DNA molecules encoding the heavy and light chain genes of the antibodies of the invention are obtained by chemical synthesis or PCR amplification. The resulting DNA molecule is inserted into an expression vector and then transfected into a host cell. Then, the transfected host cells are cultured under specific conditions and express the antibodies of the present invention.
  • Antigen-binding fragments of the present invention can be obtained by hydrolysis of intact antibody molecules (see Morimoto et al., J. Biochem. Biophys. Methods 24:107-117 (1992) and Brennan et al., Science 229:81 (1985)) .
  • these antigen-binding fragments can also be produced directly by recombinant host cells (reviewed in Hudson, Curr. Opin. Immunol. 11:548-557 (1999); Little et al., Immunol. Today, 21:364-370 (2000) )).
  • Fab' fragments can be obtained directly from host cells; Fab' fragments can be chemically coupled to form F(ab') 2 fragments (Carter et al., Bio/Technology, 10:163-167 (1992)).
  • Fv, Fab or F(ab') 2 fragments can also be directly isolated from recombinant host cell culture medium. Other techniques for preparing these antigen-binding fragments are well known to those of ordinary skill in the art.
  • the present invention provides an isolated nucleic acid molecule comprising nucleotides encoding an antibody or antigen-binding fragment thereof of the present invention, or a heavy chain variable region and/or a light chain variable region thereof sequence.
  • the nucleotide sequences are replaceable according to codon degeneracy, according to the art of codon degeneracy.
  • the nucleotide sequence is codon-optimized.
  • the present invention provides an isolated nucleic acid molecule comprising a nucleic acid molecule encoding an antibody heavy chain variable region, and/or a nucleic acid molecule encoding an antibody light chain variable region, wherein,
  • the nucleic acid molecule encoding the variable region of the antibody heavy chain has a sequence selected from the following: (a). As shown in any of SEQ ID NOs: 121, 123, 125, 127-129, 145-149, 154-158 a nucleotide sequence, or (b).
  • nucleotide sequence of (a) eg, having at least about 85%, 90 %, 95%, 99% or higher sequence identity, or a sequence with one or more nucleotide substitutions), or (c).
  • the nucleotide sequence described in (a) does not differ by more than A sequence of 3, 6, 15, 30 or 45 nucleotides; the nucleic acid molecule encoding the variable region of the antibody light chain has a sequence selected from the group consisting of: (d).
  • the nucleic acid molecule encoding the variable region of an antibody heavy chain has a nucleus as set forth in any of SEQ ID NOs: 121, 123, 125, 127-129, 145-149, 154-158
  • the nucleotide sequence, and the nucleic acid molecule encoding the variable region of the antibody light chain have the nucleotide sequence shown in any of SEQ ID NOs: 122, 124, 126, 130-144, 150-153, 159-161.
  • the isolated nucleic acid molecules of the invention comprise a variable encoding an antibody heavy chain as set forth in any of SEQ ID NOs: 121, 123, 125, 127-129, 145-149, 154-158
  • the present invention provides an isolated nucleic acid molecule. It comprises a nucleic acid molecule encoding an antibody heavy chain constant region, and/or a nucleic acid molecule encoding an antibody light chain constant region, wherein the nucleic acid molecule encoding an antibody heavy chain constant region has a sequence selected from the group consisting of: (a). The nucleotide sequence set forth in SEQ ID NO: 59, or (b).
  • nucleotide sequences differ by a sequence of no more than 3, 6, 15, 30 or 45 nucleotides, and/or, the nucleic acid molecule encoding the antibody light chain constant region has a sequence selected from the group consisting of: (d).
  • nucleotide sequence set forth in (d) eg, as set forth in (d). compared to a sequence having at least about 85%, 90%, 95%, 99% or greater sequence identity, or a sequence having one or more nucleotide substitutions), or (f). and (d) Said nucleotide sequences differ by no more than 3, 6, 15, 30 or 45 nucleotide sequences.
  • the nucleic acid molecule encoding an antibody heavy chain constant region has the nucleotide sequence set forth in SEQ ID NO: 59, and/or the nucleic acid molecule encoding an antibody light chain constant region has The nucleotide sequence shown in SEQ ID NO:26.
  • the isolated nucleic acid molecule of the present invention comprises a nucleotide sequence having the nucleotide sequence as set forth in SEQ ID NO: 59, which encodes an antibody heavy chain constant region, and/or comprises a nucleotide sequence having the nucleotide sequence as set forth in SEQ ID NO: 59, which encodes an antibody heavy chain constant region.
  • the present invention provides a vector (eg, a cloning vector or an expression vector) comprising the isolated nucleic acid molecule of the present invention.
  • the vectors of the present invention are, for example, plasmids, cosmids, phages, lentiviruses, and the like.
  • the vector is capable of expressing an antibody or antigen-binding fragment thereof of the invention in a subject (eg, a mammal, eg, a human).
  • the antibodies or antigen-binding fragments thereof of the invention can be used to construct chimeric antigen receptors (CARs) comprising an extracellular antigen-binding domain that specifically binds ROR1 (eg, ScFv), a transmembrane domain, and one or more intracellular T cell signaling domains.
  • an isolated nucleic acid molecule of the invention may comprise a nucleotide sequence encoding a chimeric antigen receptor that further comprises an antibody of the invention or its Nucleotide sequences of antigen-binding fragments (eg, ScFvs).
  • an isolated nucleic acid molecule of the invention encodes a chimeric antigen receptor comprising an antigen-binding fragment (eg, ScFv) of an antibody of the invention.
  • the antibodies or antigen-binding fragments thereof of the invention can be used to construct chimeric antigen receptor-modified immune cells comprising a chimeric antigen receptor (CAR) and Immune cells (eg T lymphocytes, NK cells).
  • CAR chimeric antigen receptor
  • Immune cells eg T lymphocytes, NK cells.
  • the present invention provides a host cell comprising an isolated nucleic acid molecule of the present invention or a vector of the present invention.
  • Host cells can be eukaryotic cells (e.g. mammalian cells, insect cells, yeast cells) or prokaryotic cells (e.g. E. coli).
  • Suitable eukaryotic cells include, but are not limited to, NSO cells, Vero cells, Hela cells, COS cells, CHO cells, ExpiCHO cells, HEK293 cells, Expi293 cells, BHK cells, and MDCKII cells.
  • Suitable insect cells include, but are not limited to, Sf9 cells.
  • the host cells of the invention are mammalian cells, such as CHO (eg, CHO-K1, CHO-S, CHO DXB11, ExpiCHO, CHO DG44).
  • the host cells of the present invention may be chimeric antigen receptor T cells (CAR-T).
  • the isolated nucleic acid molecule contained by the host cell may comprise a nucleotide sequence encoding a chimeric antigen receptor that further comprises a nucleotide sequence encoding the present invention
  • the nucleotide sequence of the antibody or antigen-binding fragment thereof eg, ScFv.
  • the isolated nucleic acid molecule contained by the host cell encodes a chimeric antigen receptor comprising an antigen-binding fragment (eg, ScFv) of an antibody of the invention.
  • a method for preparing an antibody of the present invention or an antigen-binding fragment thereof comprising, culturing a host cell of the present invention under conditions that allow expression of the antibody or antigen-binding fragment thereof, and extracting from the cultured The antibody or antigen-binding fragment thereof is recovered from the host cell culture.
  • Another aspect of the present invention discloses a pharmaceutical composition
  • a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof, carrier, host cell, conjugate, chimeric antigen receptor, or multispecific antibody of the present invention, and a pharmaceutically acceptable carrier and/or excipient.
  • compositions of the present invention comprise an antibody or antigen-binding fragment thereof of the present invention, and a pharmaceutically acceptable carrier and/or excipient.
  • the pharmaceutical compositions of the present invention comprise a vector or host cell of the present invention, and a pharmaceutically acceptable carrier and/or excipient.
  • the isolated nucleic acid molecule contained in the vector comprises a nucleotide sequence encoding a chimeric antigen receptor, the nucleotide sequence encoding a chimeric antigen receptor further comprising an antibody encoding the present invention or the nucleotide sequence of an antigen-binding fragment thereof (eg, ScFv);
  • the host cell comprises an isolated nucleic acid molecule or vector as previously described.
  • the isolated nucleic acid molecule encodes a chimeric antigen receptor comprising an antigen-binding fragment (eg, ScFv) of an antibody of the invention.
  • the host cells are T cells.
  • the host cell is a chimeric antigen receptor T cell (CAR-T).
  • the pharmaceutical composition may further comprise additional pharmaceutically active agents.
  • the additional pharmaceutically active agent is a medicament for the treatment of lymphoma, breast, colorectal, ovarian, lung and/or pancreatic cancer.
  • the additional pharmaceutically active agent is a drug with antitumor activity.
  • the additional pharmaceutically active agent is one of an epidermal growth factor receptor (EGFR) inhibitor, an immune checkpoint inhibitor, a B cell antigen inhibitor, a BTK inhibitor, a chemotherapeutic drug or more.
  • EGFR epidermal growth factor receptor
  • the antibody or antigen-binding fragment thereof of the invention and the additional pharmaceutically active agent are provided as separate components or as components of the same composition.
  • the antibody or antigen-binding fragment thereof of the invention and the additional pharmaceutically active agent may be administered simultaneously, separately or sequentially.
  • the pharmaceutical composition may further comprise additional pharmaceutically active agents.
  • the additional pharmaceutically active agent is selected from one or more of epidermal growth factor receptor (EGFR) inhibitors, immune checkpoint inhibitors, B cell antigen inhibitors, BTK inhibitors, and chemotherapeutic drugs.
  • EGFR epidermal growth factor receptor
  • the additional pharmaceutically active agent may be selected from one or more of: anti-PD1/PD-L1 antibody, anti-CD20 antibody, anti-EGFR antibody.
  • the antibody or antigen-binding fragment thereof, vector, host cell, conjugate, chimeric antigen receptor, or multispecific antibody in the pharmaceutical composition of the invention is sufficient to:
  • Another aspect of the present invention provides the use of the antibody or antigen-binding fragment thereof, carrier, host cell, conjugate, chimeric antigen receptor, or multispecific antibody of the present invention in the preparation of a medicament for:
  • the isolated nucleic acid molecule contained in the vector comprises a nucleotide sequence encoding a chimeric antigen receptor that encodes a chimeric antigen receptor.
  • the nucleotide sequence of the antigen receptor further comprises a nucleotide sequence encoding an antibody of the invention or an antigen-binding fragment thereof (eg, scFv);
  • the host cell comprises an isolated nucleic acid molecule or vector as previously described.
  • the isolated nucleic acid molecule encodes a chimeric antigen receptor comprising an antigen-binding fragment (eg, scFv) of an antibody of the invention.
  • the host cells are T cells.
  • the host cell is a chimeric antigen receptor T cell (CAR-T).
  • the medicament when a vector or host cell of the invention is used for the manufacture of a medicament, the medicament is used to treat a tumor in a subject (eg, a human).
  • a subject eg, a human
  • the antibody or antigen-binding fragment thereof, vector, host cell, conjugate, chimeric antigen receptor, or tumor involved in the multispecific antibody of the present invention is selected from lymphoma, breast cancer, colorectal cancer, ovarian cancer, lung cancer and/or pancreatic cancer.
  • the antibodies or antigen-binding fragments thereof of the present invention relate to tumors selected from lymphoma, breast cancer, colorectal cancer, ovarian cancer, lung cancer and/or pancreatic cancer.
  • the tumor involved in the antibody or antigen-binding fragment thereof of the present invention is ROR1 positive.
  • the present invention provides a method of preventing and/or treating a tumor in a subject. In another aspect, the present invention provides a method of delaying tumor progression in a subject. In another aspect, the present invention provides a method of reducing or inhibiting tumor recurrence in a subject.
  • the methods described above comprise administering to a subject in need thereof an effective amount of an antibody or antigen-binding fragment thereof, vector, host cell, conjugate, chimeric antigen receptor, or multispecific antibody of the present invention, pharmaceutical composition.
  • the invention provides a method of preventing and/or treating a tumor in a subject, comprising administering to a subject in need thereof an effective amount of an antibody or antigen-binding thereof of the invention Fragments, vectors, host cells, conjugates, chimeric antigen receptors, or multispecific antibodies, and additional pharmaceutically active agents; the additional pharmaceutically active agents may be administered separately, in combination, simultaneously, or sequentially.
  • the additional pharmaceutically active agent is selected from the group consisting of epidermal growth factor receptor (EGFR) inhibitors, immune checkpoint inhibitors, B cell antigen inhibitors, BTK inhibitors, chemotherapeutic drugs one or more.
  • EGFR epidermal growth factor receptor
  • the additional pharmaceutically active agent may be selected from one or more of: anti-PD1/PD-L1 antibody, anti-CD20 antibody, anti-EGFR antibody.
  • the host cell expresses a chimeric antigen receptor comprising an antigen-binding fragment (eg, ScFv) of an antibody of the present invention.
  • the isolated nucleic acid molecule contained in the host cell comprises a nucleotide sequence encoding a chimeric antigen receptor, the nucleotide sequence encoding a chimeric antigen receptor further comprising Nucleotide sequences encoding antibodies of the invention or antigen-binding fragments thereof (eg, ScFv).
  • the isolated nucleic acid molecule encodes a chimeric antigen receptor comprising an antigen-binding fragment (eg, ScFv) of an antibody of the invention.
  • the host cells are T cells.
  • the host cell is a chimeric antigen receptor T cell (CAR-T).
  • the above method further comprises administering to the subject a second therapy selected from the group consisting of surgery, chemotherapy, radiotherapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, Virotherapy, adjuvant therapy, and any combination thereof.
  • a second therapy selected from the group consisting of surgery, chemotherapy, radiotherapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, Virotherapy, adjuvant therapy, and any combination thereof.
  • the second therapy may be administered separately or in combination with the methods described above; or, the second therapy may be administered simultaneously or sequentially, separately or in combination with the methods described above.
  • the antibodies or antigen-binding fragments thereof, vectors, host cells, conjugates, chimeric antigen receptors or modified immune cells thereof, or multispecific antibodies of the present invention are involved in tumors Selected from lymphoma, breast cancer, colorectal cancer, ovarian cancer, lung cancer and/or pancreatic cancer.
  • the antibodies or antigen-binding fragments thereof of the present invention relate to tumors selected from lymphoma, breast cancer, colorectal cancer, ovarian cancer, lung cancer and/or pancreatic cancer.
  • the tumor involved in the antibody or antigen-binding fragment thereof of the present invention is ROR1 positive.
  • the antibodies of the present invention or antigen-binding fragments thereof, and the pharmaceutical compositions of the present invention can be formulated into any dosage form known in the medical field, for example, tablets, pills, suspensions, emulsions, solutions, gels, capsules, powders , granules, elixirs, lozenges, suppositories, injections (including injections, sterile powders for injection and concentrated solutions for injection), inhalants, sprays, etc.
  • the preferred dosage form depends on the intended mode of administration and therapeutic use.
  • the pharmaceutical compositions of the present invention should be sterile and stable under the conditions of manufacture and storage.
  • a preferred dosage form is an injection.
  • Such injectable preparations can be sterile injectable solutions.
  • Sterile injectable solutions can be prepared, for example, by incorporating the required amount of a recombinant protein of the invention in the appropriate solvent, optionally with the desired other ingredients including, but not limited to, pH adjustment agents, surfactants, adjuvants, ionic strength enhancers, isotonicity agents, preservatives, diluents, or any combination thereof) followed by filter sterilization.
  • sterile injectable solutions can be prepared as sterile lyophilized powders (eg, by vacuum drying or freeze-drying) for ease of storage and use. Such sterile lyophilized powders can be dispersed in a suitable vehicle, eg, sterile pyrogen-free water, before use.
  • antibodies or antigen-binding fragments thereof of the invention may be presented in a pharmaceutical composition in unit dosage form for ease of administration.
  • the antibodies or antigen-binding fragments thereof, pharmaceutical compositions of the present invention can be administered by any suitable method known in the art, including, but not limited to, oral, buccal, sublingual, ocular, topical, parenteral, rectal, intrathecal , intracytoplasmic reticulum, inguinal, intravesical, topical (eg, powder, ointment, or drops), or nasal route.
  • the preferred route/mode of administration is parenteral (eg, intravenous, subcutaneous, intraperitoneal, intramuscular).
  • the route and/or mode of administration will vary depending on the intended purpose.
  • the antibody or antigen-binding fragment thereof, pharmaceutical composition of the invention is administered by intravenous infusion or injection.
  • compositions of the present invention may include a "therapeutically effective amount” or a “prophylactically effective amount” of an antibody or antigen-binding fragment thereof of the present invention.
  • a “prophylactically effective amount” refers to an amount sufficient to prevent, prevent, or delay the onset of a disease.
  • a “therapeutically effective amount” refers to an amount sufficient to cure or at least partially arrest the disease and its complications in a patient already suffering from the disease.
  • the therapeutically effective amount of the antibody or antigen-binding fragment thereof of the present invention may vary according to the following factors: the severity of the disease to be treated, the general state of the patient's own immune system, the general condition of the patient such as age, weight and sex, the drug's mode of administration, and other treatments administered concurrently, among others.
  • the dosing regimen can be adjusted to obtain the optimal response of interest (eg, a therapeutic or prophylactic response). For example, a single dose may be administered, multiple doses may be administered over a period of time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation.
  • the optimal response of interest eg, a therapeutic or prophylactic response.
  • a single dose may be administered, multiple doses may be administered over a period of time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation.
  • the subject may be a mammal, such as a human.
  • the antibodies or antigen-binding fragments thereof of the present invention can specifically bind to ROR1, and thus can be used to detect the presence or level of ROR1 in a sample.
  • the present invention provides a kit comprising an antibody or antigen-binding fragment thereof of the present invention.
  • the antibodies or antigen-binding fragments thereof of the invention are detectably labeled.
  • the kit further comprises a second antibody that specifically recognizes the antibody or antigen-binding fragment thereof of the invention.
  • the second antibody further comprises a detectable label.
  • the detectable label may be any substance detectable by fluorescent, spectroscopic, photochemical, biochemical, immunological, electrical, optical or chemical means. It is particularly preferred that such labels be suitable for use in immunological detection (eg, enzyme-linked immunoassays, radioimmunoassays, fluorescent immunoassays, chemiluminescence immunoassays, etc.).
  • immunological detection eg, enzyme-linked immunoassays, radioimmunoassays, fluorescent immunoassays, chemiluminescence immunoassays, etc.
  • Such labels include, but are not limited to, enzymes (eg, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, urease, glucose oxidase, etc.), radionuclides ( For example, 3H, 125I, 35S, 14C, or 32P), fluorescent dyes (eg, fluorescein isothiocyanate (FITC), fluorescein, tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin (PE) , Texas red, rhodamine, quantum dots or cyanine derivatives (eg Cy7, Alexa 750), acridine esters, magnetic beads (eg, ), calorimetric labels such as colloidal gold or colored glass or plastic (eg, polystyrene, polypropylene, latex, etc.) beads, and modified avidins (eg, streptavidin) for binding to the aforementioned labels
  • enzymes e
  • Labels encompassed by the present invention can be detected by methods known in the art. For example, radiolabels can be detected using photographic film or a scintillation calculator, and fluorescent labels can be detected using a light detector to detect the emitted light. Enzyme labels are generally detected by providing a substrate to the enzyme and detecting the reaction product produced by the action of the enzyme on the substrate, and calorimetric labels are detected by simply visualizing the colored label. In certain embodiments, detectable labels as described above can be attached to recombinant proteins of the invention via linkers of varying lengths to reduce potential steric hindrance.
  • the invention provides a method of detecting the presence or level of ROR1 in a sample comprising the step of using an antibody or antigen-binding fragment thereof of the invention.
  • the antibody or antigen-binding fragment thereof of the invention is also detectably labeled.
  • the method further comprises using a detectably labeled reagent to detect the antibody or antigen-binding fragment thereof of the present invention.
  • the method may be used for diagnostic purposes, or for non-diagnostic purposes (eg, the sample is a cell sample rather than a sample from a patient).
  • the present invention provides a method for detecting the presence or level of ROR1 in a sample, the method comprising causing the antibody or antigen-binding fragment thereof to form a complex with ROR1 under conditions that allow the formation of a complex between the antibody or antigen-binding fragment thereof and ROR1.
  • a sample is contacted with an antibody or antigen-binding fragment thereof of the invention, and the formation of the complex is detected.
  • an antibody or antigen-binding fragment thereof of the present invention in the preparation of a kit for detecting the presence or level of ROR1 in a sample.
  • the present invention provides diagnostic or therapeutic kits comprising the antibodies or antigen-binding fragments thereof, vectors, host cells, conjugates, chimeric antigen receptors, or multispecific antibodies of the present invention Antibodies, and instructions for use.
  • tumors and tumor metastasis can be diagnosed by detecting the presence or level of ROR1 in a sample. Accordingly, the antibodies or antigen-binding fragments, conjugates, multispecific antibodies, or kits thereof of the present invention can be used to diagnose tumors and tumor metastasis. Accordingly, in another aspect, there is provided the use of an antibody or antigen-binding fragment, conjugate, multispecific antibody, or kit of the present invention in diagnosing tumors and tumor metastasis.
  • the antibody of the present invention has high binding affinity to ROR1 and has extremely strong specificity. Therefore, the antibodies of the present invention have potential for preventing and/or treating tumors.
  • the humanized antibodies of the present invention retain the functions and properties of the parental murine antibodies. Furthermore, the humanized antibodies of the present invention have a high degree of humanization, so that they can be safely administered to human subjects without eliciting an immunogenic response. Therefore, the antibodies (especially humanized antibodies) of the present invention have great clinical value.
  • FR antibody framework region amino acid residues other than CDR residues in the variable region of an antibody
  • IMGT is based on the International Information System on Immunogenetics, initiated by Lefranc et al.
  • CDR-H1 Complementarity determining region 1 in the variable region of an immunoglobulin heavy chain
  • antibody refers to an immunoglobulin molecule generally composed of two pairs of polypeptide chains, each pair having one light (LC) and one heavy (HC) chain.
  • Antibody light chains can be classified as kappa (kappa) and lambda (lambda) light chains.
  • Heavy chains can be classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively.
  • the variable and constant regions are linked by a "J" region of about 12 or more amino acids, and the heavy chain also contains a "D" region of about 3 or more amino acids.
  • Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH).
  • the heavy chain constant region consists of 3 domains (CH1, CH2 and CH3).
  • Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL).
  • the light chain constant region consists of one domain, CL.
  • Constant domains are not directly involved in the binding of antibodies to antigens, but exhibit a variety of effector functions, such as mediating immunoglobulins with host tissues or factors, including various cells of the immune system (eg, effector cells) and classical complement Binding of the first component (C1q) of the system.
  • VH and VL regions can also be subdivided into regions of high variability called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs).
  • CDRs complementarity determining regions
  • FRs framework regions
  • Each VH and VL consists of 3 CDRs and 4 FRs arranged in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from amino terminus to carboxy terminus.
  • the assignment of amino acids to regions or domains can follow Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196 : 901-917; definition by Chothia et al. (1989) Nature 342:878-883.
  • antibody includes not only whole antibodies but also antigen-binding fragments of antibodies.
  • CDR complementarity determining region
  • the term "complementarity determining region" or "CDR” refers to the amino acid residues in the variable region of an antibody that are responsible for antigen binding.
  • the precise boundaries of these amino acid residues can be defined according to various numbering systems known in the art, for example according to the AbM numbering system (Martin ACR, Cheetham JC, Rees AR (1989) Modelling antibody hypervariable loops: Combined algorithm. Proc Natl Acad Sci USA 86:9268–9272) or the IMGT numbering system (Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003).
  • the CDRs defined by each numbering system.
  • the correspondence between different numbering systems is well known to those skilled in the art (see, eg, Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003).
  • the CDRs contained in the antibody or antigen-binding fragment thereof of the present invention can be determined according to various numbering systems known in the art.
  • the CDRs contained by an antibody or antigen-binding fragment thereof of the invention are preferably determined by the Kabat, Chothia, AbM, IMGT numbering system.
  • framework region or "FR” residues refers to those amino acid residues in the variable region of an antibody other than the CDR residues as defined above.
  • “Germline antibody genes” are immunoglobulin sequences encoded by non-lymphocytes that have not undergone the maturation process that leads to genetic rearrangement and maturation of specific immunoglobulins.
  • One advantage provided by various embodiments of the present invention arises from the recognition that germline antibody genes retain more of the important amino acid sequence structure characteristic of individuals of animal species than mature antibody genes. Thus, when applied therapeutically to that species, it is less likely to be recognized as a foreign substance by that species.
  • antibody is not limited by any particular method of producing an antibody. For example, it includes recombinant antibodies, monoclonal antibodies and polyclonal antibodies. Antibodies may be of different isotypes, eg, IgG (eg, IgGl, IgG2, IgG3 or IgG4 subtype), IgAl, IgA2, IgD, IgE or IgM antibodies.
  • IgG eg, IgGl, IgG2, IgG3 or IgG4 subtype
  • IgAl IgA2, IgD, IgE or IgM antibodies.
  • the term "antigen-binding fragment" of an antibody refers to a polypeptide of a fragment of an antibody, such as a polypeptide of a fragment of a full-length antibody, that retains the ability to specifically bind the same antigen to which the full-length antibody binds, and/or Or compete with a full-length antibody for specific binding to an antigen, which is also referred to as an "antigen-binding portion.”
  • an antigen-binding portion See generally, Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd ed., Raven Press, NY (1989), which is hereby incorporated by reference in its entirety for all purposes.
  • antigen-binding fragments of antibodies are produced by enzymatic or chemical cleavage of intact antibodies.
  • Non-limiting examples of antigen-binding fragments include Fab, Fab', F(ab') 2 , Fd, Fv, dAb, and complementarity determining regions (CDRs).
  • Engineered antibody variants are reviewed in Holliger et al., 2005; Nat Biotechnol, 23: 1126-1136.
  • full-length antibody means an antibody consisting of two “full-length heavy chains” or “heavy chains” and two “full-length light chains” or “light chains.”
  • full-length heavy chain or “heavy chain” refers to such a polypeptide chain, which in the direction from N-terminus to C-terminus consists of heavy chain variable region (VH), heavy chain constant region CH1 domain, hinge region ( HR), heavy chain constant region CH2 domain, heavy chain constant region CH3 domain; and, when the full-length antibody is of the IgE isotype, optionally also includes a heavy chain constant region CH4 domain.
  • a "full-length heavy chain” is a polypeptide chain consisting of VH, CH1, HR, CH2 and CH3 in the N-terminal to C-terminal direction.
  • a "full-length light chain” or “light chain” is a polypeptide chain consisting of a light chain variable region (VL) and a light chain constant region (CL) in the N-terminal to C-terminal direction.
  • the two pairs of full-length antibody chains are linked together by a disulfide bond between CL and CH1 and a disulfide bond between the HRs of the two full-length heavy chains.
  • the full-length antibody of the present invention can be from a single species, such as human; it can also be a chimeric antibody or a humanized antibody.
  • the full-length antibodies of the present invention comprise two antigen-binding sites formed by VH and VL pairs, respectively, which specifically recognize/bind to the same antigen.
  • Fd fragment means an antibody fragment consisting of VH and CH1 domains
  • dAb fragment means an antibody fragment consisting of a VH domain (Ward et al., Nature 341:544546 (1989))
  • Fab fragment means an antibody fragment consisting of VL, VH, CL and CH1 domains
  • F(ab')2 fragment means an An antibody fragment of two Fab fragments
  • Fab' fragment means a fragment obtained by reducing the disulfide bond linking two heavy chain fragments in an F(ab')2 fragment, consisting of an intact light chain and a heavy chain. Fd fragment (consisting of VH and CH1 domains).
  • Fv fragment means an antibody fragment consisting of the one-armed VL and VH domains of an antibody. Fv fragments are generally considered to be the smallest antibody fragments that can form a complete antigen-binding site. It is generally believed that the six CDRs confer antigen-binding specificity to an antibody. However, even a single variable region (eg, an Fd fragment, which contains only three CDRs specific for the antigen) is able to recognize and bind the antigen, albeit probably with lower affinity than the intact binding site.
  • Fc fragment means that the second and third constant regions of the first heavy chain of an antibody are disulfide bonded to the second and third constant regions of the second heavy chain. antibody fragments.
  • the Fc fragment of an antibody has many different functions, but is not involved in antigen binding.
  • scFv refers to a single polypeptide chain comprising VL and VH domains, wherein the VL and VH are connected by a linker (see, e.g., Bird et al., Science 242:423 -426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Eds. Roseburg and Moore, Springer-Verlag, New York, pp. 269-315 (1994)).
  • Such scFv molecules can have the general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH.
  • Suitable prior art linkers consist of repeated GGGGS amino acid sequences or variants thereof.
  • GGGGS linker with the amino acid sequence
  • Other linkers useful in the present invention are described by Alfthan et al. (1995), Protein Eng. 8:725-731, Choi et al. (2001), Eur. J. Immunol. 31:94-106, Hu et al. (1996), Cancer Res.
  • a disulfide bond may also exist between the VH and VL of the scFv.
  • the term "di-scFv" refers to an antibody fragment formed by the joining of two scFvs.
  • the term "diabody” means that the VH and VL domains of which are expressed on a single polypeptide chain, but the use of linkers that are too short to allow between the two domains of the same chain inter-pairing, thereby forcing the domains to pair with the complementary domains of the other chain and creating two antigen-binding sites (see, e.g., Holliger P. et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993) ), and Poljak R.J. et al., Structure 2:1121-1123 (1994)).
  • Each of the aforementioned antibody fragments retains the ability to specifically bind to the same antigen bound by the full-length antibody, and/or compete with the full-length antibody for specific binding to the antigen.
  • multispecific antibody refers to antibodies having a variety of different antigen-binding specificities, including, for example, bispecific antibodies, trispecific antibodies, and tetraspecific antibodies.
  • Bispecific antibody refers to an antibody with two different antigen-binding specificities, which is formed by a first antibody (or fragment thereof) and a second antibody (or fragment thereof) or antibody analog through a coupling arm Conjugate, the way of conjugation includes but is not limited to chemical reaction, gene fusion and enzymatic.
  • Multispecific antibodies include, for example, trispecific antibodies, which are antibodies with three different antigen-binding specificities, and tetraspecific antibodies, which are antibodies with four different antigen-binding specificities Antibody.
  • antibody mimetics refers to the same specificity as an antibody that binds to an antigen, but lacks the antibody structure. They are usually artificial peptides or proteins with a molar mass of about 3 to 20 kDa.
  • DARPins ankyrin repeat proteins
  • fynomers Designed ankyrin repeat protein (DARPin) linked to IgG antibody, scFv-Fc antibody fragment or a combination thereof, eg CN104341529A.
  • Anti-IL-17a fynomers bind to anti-IL-6R antibodies, as in WO2015141862A1.
  • CAR Chimeric Antigen Receptor
  • immunoglobulin can refer to a class of proteins that function as antibodies. Antibodies expressed by B cells are sometimes called chimeric antigen receptors or antigen receptors. The five members included in this class of proteins are IgA, IgG, IgM, IgD and IgE, with IgG being the most common circulating antibody. It is the most potent immunoglobulin in agglutination, complement fixation and other antibody responses and is important in defense against bacteria and viruses.
  • techniques for obtaining antibodies can use conventional techniques known to those of skill in the art (eg, recombinant DNA techniques or enzymatic or chemical cleavage methods) to obtain the antigens of the antibodies from a given antibody (eg, the antibodies provided herein) Fragments (eg, the antibody fragments described above) are bound, and the antibody is screened for specificity for antigen-binding fragments in the same manner as is used for intact antibodies.
  • a given antibody eg, the antibodies provided herein
  • Fragments eg, the antibody fragments described above
  • the terms "monoclonal antibody”, “monoclonal antibody”, “mAb” have the same meaning and are used interchangeably and interchangeably, and refer to one from a population of highly homologous antibody molecules An antibody or a fragment of an antibody, that is, a population of identical antibody molecules, except for natural mutations that may arise spontaneously.
  • Monoclonal antibodies are highly specific for a single epitope on an antigen.
  • Polyclonal antibodies are relative to monoclonal antibodies, which generally comprise at least two or more different antibodies that generally recognize different epitopes on an antigen.
  • the modifier "monoclonal” only indicates that the antibody is characterized as being obtained from a population of highly homologous antibodies and should not be construed as requiring any particular method to prepare said antibody.
  • Monoclonal antibodies of the invention can be prepared by a variety of techniques, such as hybridoma technology (see, eg, Kohler et al. Nature, 256:495, 1975), recombinant DNA technology (see, eg, US Patent Application 4,816,567), or bacteriophage Antibody library technology (see, eg, Clackson et al. Nature 352:624-628, 1991, or Marks et al. J. Mol. Biol. 222:581-597, 1991).
  • monoclonal antibodies can be prepared as follows. Mice or other suitable host animals are first immunized with the immunogen (adjuvanted if necessary).
  • the immunogen or adjuvant is usually injected subcutaneously at multiple points or intraperitoneally.
  • the immunogen can be preconjugated to certain known proteins, such as serum albumin or soybean trypsin inhibitor, to enhance the immunogenicity of the antigen in the host.
  • the adjuvant may be Freund's adjuvant or MPL-TDM or the like.
  • lymphocytes can also be obtained by in vitro immunization.
  • Lymphocytes of interest are collected and fused with myeloma cells using a suitable fusion agent, such as PEG, to obtain hybridoma cells (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103, Academic Press, 1996).
  • a suitable fusion agent such as PEG
  • the hybridoma cells prepared above can be inoculated into a suitable culture medium for growth, and the culture medium preferably contains one or more substances capable of inhibiting the growth of unfused, parental myeloma cells.
  • hypoxanthine guanine phosphotransferase HGPRT or HPRT
  • HAT medium hypoxanthine guanine phosphotransferase
  • the preferred myeloma cells should have the characteristics of high fusion rate, stable antibody secretion ability, and sensitivity to HAT medium.
  • murine myeloma cells are preferred, such as MOP-21 or MC-11 mouse tumor-derived strains (THE Salk Institute Cell Distribution Center, San Diego, Calif. USA), and SP-2/0 or X63-Ag8 -653 cell line (American Type Culture Collection, Rockville, Md. USA).
  • Methods for determining the binding specificity of monoclonal antibodies produced by hybridoma cells include, for example, immunoprecipitation or in vitro binding assays such as radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA).
  • RIA radioimmunoassay
  • ELISA enzyme-linked immunosorbent assay
  • the affinity of mAbs can be determined using the Scatchard assay described by Munson et al., Anal. Biochem. 107:220 (1980). After the specificity, affinity and reactivity of the antibodies produced by the hybridomas have been determined, the cell line of interest can pass the limited criteria described in (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103, Academic Press, 1996). Subcloning by dilution.
  • a suitable medium can be DMEM or RPMI-1640 and the like.
  • hybridoma cells can also grow in animals in the form of ascites tumors.
  • immunoglobulin purification methods such as protein A agarose gel, hydroxyapatite chromatography, gel electrophoresis, dialysis or affinity chromatography, the monoclonal antibodies secreted by subcloned cells can be purified from cell culture medium, ascites or serum.
  • Monoclonal antibodies can also be obtained by genetic engineering recombinant technology. Using nucleic acid primers that specifically bind to the heavy chain and light chain genes of the monoclonal antibody to perform PCR amplification, the DNA molecules encoding the heavy chain and light chain genes of the monoclonal antibody can be isolated from the hybridoma cells. Insert the obtained DNA molecule into an expression vector, then transfect host cells (such as E. coli cells, COS cells, CHO cells, or other myeloma cells that do not produce immunoglobulins), and culture under suitable conditions, Recombinantly expressed antibodies of interest can be obtained.
  • host cells such as E. coli cells, COS cells, CHO cells, or other myeloma cells that do not produce immunoglobulins
  • Antibodies can be purified by well-known techniques, such as affinity chromatography using protein A or protein G. Subsequently or alternatively, the specific antigen (the target molecule recognized by the antibody) or its epitope can be immobilized on a column and the immunospecific antibody purified by immunoaffinity chromatography.
  • immunoaffinity chromatography For the purification of immunoglobulin, refer to, for example, D. Wilkinson (The Engineer, published by The Engineer, Inc., Philadelphia Pa., Vol. 14, No. 8 (Apr. 17, 2000), pp. 25-28).
  • murine antibody refers to the fusion of B cells derived from an immunized mouse with myeloma cells, followed by screening for murine hybrid fusion cells that can both proliferate immortally and secrete antibodies, Further screening, antibody preparation and antibody purification were performed. Or due to the differentiation and proliferation of B cells after the antigen invades the mouse body, plasma cells are formed, and plasma cells can produce and secrete antibodies. Stimulated by a specific antigen, the production of antibodies is due to the interaction of various immune cells after the antigen invades the human body, so that the B cells in the lymphocytes differentiate and proliferate to form plasma cells, which can produce and secrete antibodies.
  • chimeric antibody refers to an antibody whose light chain or/and a portion of its heavy chain is derived from an antibody (which may be derived from a particular species or belong to a a particular antibody class or subclass), and another portion of the light chain or/and heavy chain is derived from another antibody (which may be derived from the same or a different species or of the same or different antibody class or subclass), but regardless of whether the However, it still retains binding activity to the target antigen (U.S.P 4,816,567 to Cabilly et al.; Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851 6855 (1984)).
  • chimeric antibody can include antibodies (eg, human-mouse chimeric antibodies) in which the heavy and light chain variable regions of the antibody are derived from a primary antibody (eg, a murine antibody) and the heavy and The light chain constant region is derived from a second antibody (eg, a human antibody).
  • a primary antibody eg, a murine antibody
  • a second antibody eg, a human antibody
  • humanized antibody refers to a genetically engineered non-human antibody whose amino acid sequence has been modified to increase homology to the sequence of a human antibody.
  • CDR regions of a humanized antibody are derived from a non-human antibody (donor antibody), and all or part of the non-CDR regions (eg, variable FR and/or constant regions) are derived from human Immunoglobulins (receptor antibodies).
  • Humanized antibodies generally retain the expected properties of the donor antibody, including, but not limited to, antigen specificity, affinity, reactivity, the ability to increase immune cell activity, the ability to enhance immune responses, and the like.
  • the donor antibody can be a mouse, rat, rabbit, or non-human primate with the desired properties (eg, antigen specificity, affinity, reactivity, ability to increase immune cell activity and/or ability to enhance immune response) Animal-like (eg, cynomolgus monkey) antibodies.
  • Humanized antibodies can both retain the expected properties of non-human donor antibodies (such as murine antibodies), and can effectively reduce the immunogenicity of non-human donor antibodies (such as murine antibodies) in human subjects, Therefore, it is particularly advantageous.
  • the expected properties of the humanized antibody eg, antigen specificity, affinity, reactivity, ability to enhance immune cell activity, and/or ability to enhance the immune response
  • Technologists need to explore, explore and transform specific donor antibodies, and only after a lot of creative work can they be obtained, which have a high degree of humanization (for example, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% degree of humanization), while retaining the expectation of a specific donor antibody humanized antibodies.
  • a high degree of humanization for example, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% degree of humanization
  • the framework regions (FRs) of the humanized antibodies of the present invention may include both the amino acid residues of the human acceptor antibody and the amino acid residues of the corresponding non-human donor antibody.
  • the chimeric antibody or humanized antibody of the present invention can be prepared according to the sequence of the mouse monoclonal antibody prepared above.
  • DNA encoding the heavy and light chains can be obtained from target murine hybridomas and engineered to contain non-murine (eg, human) immunoglobulin sequences using standard molecular biology techniques.
  • murine immunoglobulin variable regions can be linked to human immunoglobulin constant regions using methods known in the art (see, eg, US Patent No. 4,816,567 to Cabilly et al.).
  • the VH-encoding DNA is operably linked to another DNA molecule encoding the heavy chain constant region to obtain a full-length heavy chain gene.
  • the sequences of human heavy chain constant region genes are known in the art (see, e.g., Kabat, E.A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242 ), DNA fragments containing these regions can be obtained by standard PCR amplification.
  • the heavy chain constant region may be an IgGl, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region, but is generally preferably an IgGl or IgG4 constant region.
  • the DNA encoding VL is operably linked to another DNA molecule encoding the light chain constant region CL to obtain a full-length light chain gene (as well as a Fab light chain gene).
  • Sequences of human light chain constant region genes are known in the art (see, e.g., Kabat, E.A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242 ), DNA fragments containing these regions can be obtained by standard PCR amplification.
  • the light chain constant region may be a kappa or lambda constant region, but is generally preferably a kappa constant region.
  • murine CDR regions can be inserted into human framework sequences using methods known in the art (see US Patent Nos. 5,225,539 to Winter; US Patent Nos. 5,530,101 to Queen et al; 5,585,089; 5,693,762 and 6,180,370; and Lo, Benny, K.C., editor, in Antibody Engineering: Methods and Protocols, volume 248, Humana Press, New Jersey, 2004).
  • transgenic animals that are capable of producing no endogenous immunoglobulins following immunization and capable of producing fully human antibody repertoires can also be utilized.
  • JH antibody heavy chain joining region
  • Non-limiting examples of such transgenic animals include, HuMAb mice (Medarex, Inc.), which contain human immunoglobulin gene miniatures encoding unrearranged human heavy (mu and gamma) and kappa light chain immunoglobulin sequences. loci (miniloci), plus targeted mutations that inactivate endogenous mu and kappa chain loci (see, eg, Lonberg et al. (1994) Nature 368(6474):856-859); or carrying a human heavy chain transgene and human "KM mouse TM" of the light chain transchromosome (see patent application WO02/43478).
  • Other methods for humanizing antibodies include phage display technology (Hoogenboom et al., 1991, J. Mol. Biol. 227:381; Marks et al., J. Mol. Biol. 1991, 222:581-597; Vaughan et al., 1996 , Nature Biotech 14:309).
  • degree of humanization is an indicator used to evaluate the number of non-human amino acid residues in a humanized antibody.
  • the degree of humanization of a humanized antibody can be predicted, for example, by using the IMGT website DomainGapAlign to predict the homology of the variable region sequence to the human V domain.
  • homologous antibodies refer to variants of antibodies comprising heavy and light chain variable regions comprising amino acid sequences identical to the amino acid sequences of the antibodies or antigen-binding fragments thereof provided herein source, and wherein the variant retains the desired functional properties of the anti-ROR1 antibodies of the invention.
  • Sequence alignment methods for comparison are well known in the art. Various procedures and alignment algorithms are described in: Smith TF and Waterman MS, Adv. Appl. Math., 2:482, 1981; Higgins DG and Sharp PM, CABIOS 5:151, 1989. Altschul SF et al., Nature Genet., 6:119, 1994 provide detailed ideas for sequence alignment methods and homology calculations.
  • telomere binding refers to a non-random binding reaction between two molecules, such as between an antibody and the antigen to which it is directed.
  • the strength or affinity of a specific binding interaction can be expressed as the equilibrium dissociation constant (KD) or half-maximal effect concentration (EC50) for that interaction.
  • the specific binding properties between two molecules can be determined using methods well known in the art.
  • One method involves measuring the rate of antigen binding site/antigen complex formation and dissociation.
  • Both the "association rate constant” (ka or kon) and the “dissociation rate constant” (kdis or koff) can be calculated from the concentrations and the actual rates of association and dissociation (see Malmqvist M, Nature, 1993, 361 :186-187).
  • the ratio of kdis/kon is equal to the dissociation constant KD (see Davies et al., Annual Rev Biochem, 1990;59:439-473).
  • KD, kon and kdis values can be measured by any valid method.
  • dissociation constants can be measured using bioluminescence interferometry (eg, the ForteBio Octet method). In addition to this, dissociation constants can also be measured using surface plasmon resonance techniques (eg Biacore) or Kinexa.
  • the term "vector” refers to a nucleic acid delivery vehicle into which a polynucleotide can be inserted.
  • the vector can express the protein encoded by the inserted polynucleotide, the vector is called an expression vector.
  • the vector can be introduced into a host cell by transformation, transduction or transfection, so that the genetic material elements carried by it can be expressed in the host cell.
  • Vectors are well known to those skilled in the art and include, but are not limited to: plasmids; phagemids; cosmids; artificial chromosomes, such as yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs) or P1 derived artificial chromosomes (PACs) ; Phage such as ⁇ phage or M13 phage and animal viruses.
  • YACs yeast artificial chromosomes
  • BACs bacterial artificial chromosomes
  • PACs P1 derived artificial chromosomes
  • Animal viruses that can be used as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpesviruses (eg, herpes simplex virus), poxviruses, baculoviruses, papillomaviruses, papillomaviruses Polyoma vacuolar virus (eg SV40).
  • retroviruses including lentiviruses
  • adenoviruses eg, adeno-associated viruses
  • herpesviruses eg, herpes simplex virus
  • poxviruses baculoviruses
  • papillomaviruses papillomaviruses
  • Polyoma vacuolar virus eg SV40
  • a vector may contain a variety of elements that control expression, including, but not limited to, promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and
  • Expression and cloning vectors contain nucleic acid sequences that enable the vector to replicate in one or more host cells of choice. Typically, in cloning vectors, this sequence is what enables the vector to replicate independently of the host chromosomal DNA, and it includes an origin of replication or an autonomously replicating sequence.
  • expression vector refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operably linked to the nucleotide sequence to be expressed. Expression vectors contain sufficient cis-acting elements for expression; other elements for expression can be provided by host cells or in vitro expression systems.
  • Expression vectors include all those known in the art, such as cosmids, plasmids (eg, naked or contained in liposomes), and viruses (eg, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses).
  • cosmids eg, naked or contained in liposomes
  • viruses eg, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses.
  • the term "host cell” refers to a cell that can be used to introduce a vector, including, but not limited to, prokaryotic cells such as E. coli or Bacillus subtilis, fungal cells such as yeast cells or Aspergillus, etc., Insect cells such as S2 Drosophila cells or Sf9, or insect cells such as fibroblasts, NSO cells, Vero cells, Hela cells, COS cells, CHO cells (e.g. CHO-K1, CHO-S, CHO DXB11, ExpiCHO, CHO DG44 cells ), ExpiCHO cells, HEK293 cells, Expi293 cells, BHK cells, and MDCKII cells and other animal cells.
  • host cells also include immune cells used to construct chimeric antigen receptor T cells (CAR-T), such as T lymphocytes, NK cells, etc.
  • CAR-T chimeric antigen receptor T cells
  • identity is used to refer to the match of sequences between two polypeptides or between two nucleic acids.
  • a position in both sequences being compared is occupied by the same base or amino acid monomer subunit (e.g., a position in each of two DNA molecules is occupied by an adenine, or both A position in each of the polypeptides is occupied by a lysine)
  • the molecules are identical at that position.
  • the "percent identity” between two sequences is a function of the number of matched positions shared by the two sequences divided by the number of positions compared x 100. For example, two sequences are 60% identical if 6 out of 10 positions match.
  • the DNA sequences CTGACT and CAGGTT share 50% identity (matching at 3 positions out of a total of 6).
  • comparisons are made when two sequences are aligned for maximum identity.
  • Such alignment can be accomplished using, for example, the method of Needleman et al. (1970) J. Mol. Biol. 48:443-453, which can be conveniently performed by a computer program such as the Align program (DNAstar, Inc.).
  • Align program DNAstar, Inc.
  • Appl Biosci., 4:11-17 (1988)) integrated into the ALIGN program (version 2.0) can also be used, using the PAM120 weight residue table , a gap length penalty of 12, and a gap penalty of 4 to determine the percent identity between two amino acid sequences.
  • the algorithm of Needleman and Wunsch (J MoI Biol. 48:444-453 (1970)) in the GAP program integrated into the GCG software package (available at www.gcg.com), using the Blossum 62 matrix or PAM250 matrix with gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6 to determine percent identity between two amino acid sequences .
  • conservative substitutions means amino acid substitutions that do not adversely affect or alter the intended properties of the protein/polypeptide comprising the amino acid sequence.
  • conservative substitutions can be introduced by standard techniques known in the art such as site-directed mutagenesis and PCR-mediated mutagenesis.
  • Conservative amino acid substitutions include substitutions of amino acid residues with amino acid residues that have similar side chains, e.g., that are physically or functionally similar to the corresponding amino acid residues (e.g., have similar size, shape, charge, chemical properties, including the ability to form covalent bonds or hydrogen bonds, etc.) Families of amino acid residues with similar side chains have been defined in the art.
  • These families include those with basic side chains (eg, lysine, arginine, and histidine), acidic side chains (eg, aspartic acid, glutamic acid), uncharged polar side chains (eg, glycine) , asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), non-polar side chains (e.g.
  • alanine, valine, leucine, isoleucine amino acid, proline, phenylalanine, methionine), beta branched side chains (eg, threonine, valine, isoleucine), and aromatic side chains (eg, tyrosine, phenylalanine, tryptophan, histidine). Therefore, it is preferred to replace the corresponding amino acid residue with another amino acid residue from the same side chain family.
  • Methods for identifying conservative substitutions of amino acids are well known in the art (see, eg, Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al. Protein Eng. 12(10):879-884 (1999) and Burks et al. Proc. Natl Acad. Set USA 94:412-417 (1997), which is incorporated herein by reference).
  • amino acids are generally represented by one-letter and three-letter abbreviations well known in the art.
  • alanine can be represented by A or Ala.
  • the term "pharmaceutically acceptable carrier and/or excipient” refers to a carrier and/or excipient that is pharmacologically and/or physiologically compatible with the subject and the active ingredient, It is well known in the art (see e.g. Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995) and includes, but is not limited to: pH adjusters, surfactants, adjuvants, ionic strength enhancers Agents, diluents, agents to maintain osmotic pressure, agents to delay absorption, preservatives.
  • pH adjusting agents include, but are not limited to, phosphate buffers.
  • Surfactants include, but are not limited to, cationic, anionic or nonionic surfactants, such as Tween-80.
  • Ionic strength enhancers include, but are not limited to, sodium chloride.
  • Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and the like.
  • Agents for maintaining osmotic pressure include, but are not limited to, sugars, NaCl, and the like.
  • Agents that delay absorption include, but are not limited to, monostearate salts and gelatin.
  • Diluents include, but are not limited to, water, aqueous buffers (eg, buffered saline), alcohols and polyols (eg, glycerol), and the like.
  • Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid, and the like.
  • Stabilizers have the meaning commonly understood by those skilled in the art, which are capable of stabilizing the desired activity of the active ingredient in the drug, including but not limited to sodium glutamate, gelatin, SPGA, sugars (such as sorbitol, mannitol, starch, sucrose , lactose, glucan, or glucose), amino acids (such as glutamic acid, glycine), proteins (such as dry whey, albumin or casein) or their degradation products (such as lactalbumin hydrolyzate) and the like.
  • sugars such as sorbitol, mannitol, starch, sucrose , lactose, glucan, or glucose
  • amino acids such as glutamic acid, glycine
  • proteins such as dry whey, albumin or casein
  • degradation products such as lactalbumin hydrolyzate
  • prevention refers to a method performed to prevent or delay the occurrence of a disease or disorder or symptom (e.g., a tumor) in a subject.
  • treatment refers to a method performed to obtain a beneficial or desired clinical result.
  • a beneficial or desired clinical outcome includes, but is not limited to, alleviation of symptoms, reduction in the extent of the disease, stabilization (ie, not worsening) of the disease state, delaying or slowing the progression of the disease, amelioration or alleviation of the disease status, and relief of symptoms (whether in part or in full), whether detectable or undetectable.
  • treatment can also mean prolonging survival as compared to expected survival if not receiving treatment.
  • the term "subject” refers to a mammal, such as a primate, such as a human.
  • the subject eg, human
  • the term "effective amount" refers to an amount sufficient to obtain, or at least partially obtain, the desired effect.
  • a disease-prophylactically effective amount refers to an amount sufficient to prevent, arrest, or delay the onset of a disease (eg, a tumor);
  • a disease-treatment effective amount refers to an amount sufficient to cure or at least partially prevent an existing disease
  • an amount effective for therapeutic use will depend on the severity of the disease to be treated, the general state of the patient's own immune system, the patient's general condition such as age, weight and sex, the mode of administration of the drug, and other concurrently administered treatments and many more.
  • effector function refers to those biological activities attributable to an Fc region of an antibody (either a native sequence Fc region or an amino acid sequence variant Fc region), and which vary with the antibody Isotypes vary.
  • antibody effector functions include, but are not limited to: Fc receptor binding affinity, antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), antibody-dependent cellular phagocytosis (ADCP) , downregulation of cell surface receptors (eg B cell receptors), B cell activation, cytokine secretion, half-life/clearance of antibodies and antigen-antibody complexes, etc.
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • CDC complement-dependent cytotoxicity
  • ADCP antibody-dependent cellular phagocytosis
  • downregulation of cell surface receptors eg B cell receptors
  • B cell activation eg B cell activation
  • cytokine secretion half-life/clearance of antibodies and antigen-antibody complexe
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • Ig interacts with cytotoxic cells (eg, natural killer (NK) cells, neutrophils Fc receptors (FcR) present on macrophages) bind to these cytotoxic effector cells to specifically bind to antigen-attached target cells, and then kill the target cells by secreting cytotoxins.
  • cytotoxic cells eg, natural killer (NK) cells, neutrophils Fc receptors (FcR) present on macrophages
  • FcR neutrophils Fc receptors
  • complement-dependent cytotoxicity refers to a form of cytotoxicity that activates the complement cascade by binding complement component C1q to an antibody Fc.
  • Methods for detecting the CDC activity of an antibody are known in the art, and can be assessed, for example, by measuring the binding activity between the antibody to be tested and an Fc receptor (eg, C1q).
  • the term "pharmaceutically acceptable” means that the molecular entity, molecular fragment or composition does not produce adverse, allergic or other untoward reactions when properly administered to an animal or human.
  • pharmaceutically acceptable carriers or components thereof include sugars (eg, lactose), starch, cellulose and derivatives thereof, vegetable oils, gelatin, polyols (eg, propylene glycol), alginic acid, and the like.
  • combination therapy includes combining the present invention with an anti-ROR1 antibody or antigen-binding fragment thereof with one or more additional active therapeutic agents (eg, chemotherapeutic agents) or other prophylactic or therapeutic modalities (eg, radiation therapy) used in combination.
  • additional active therapeutic agents eg, chemotherapeutic agents
  • other prophylactic or therapeutic modalities eg, radiation therapy
  • Combination therapy includes therapeutic agents that affect the immune response (eg, enhance or activate the response) and therapeutic agents that affect (eg, inhibit or kill) the tumor/cancer cells. Combination therapy reduces the likelihood of drug-resistant cancer cells developing. Combination therapy may allow for a dose reduction of one or more of the agents to reduce or eliminate adverse effects associated with one or more of the agents. Such combination therapy may have a synergistic therapeutic or prophylactic effect on the underlying disease, disorder or condition.
  • “combination” includes therapies that can be administered separately, eg, formulated separately for separate administration (eg, can be provided in a kit), and therapies that can be administered together in a single formulation (ie, a "co-formulation") .
  • the anti-ROR1 antibodies or antigen-binding fragments thereof of the invention can be administered sequentially.
  • the anti-ROR1 antibodies or antigen-binding fragments thereof can be administered concurrently.
  • the anti-ROR1 antibodies or antigen-binding fragments thereof of the invention can be used in any combination with at least one other (active) agent.
  • ROR1 positive was obtained by immunohistochemistry and staining intensity evaluation by professional clinicopathologists.
  • cancer and “tumor” are used interchangeably and refer to a large group of diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division can lead to the formation of malignant tumors or cells that invade adjacent tissues and may metastasize to distant sites in the body through the lymphatic system or bloodstream. Cancers include benign and malignant cancers as well as dormant tumors or micrometastases. Cancer also includes hematological malignancies.
  • lymphoma includes lymphoma, leukemia, myeloma or lymphoid malignancies, as well as spleen and lymph node tumors.
  • exemplary lymphomas include B-cell lymphomas and T-cell lymphomas.
  • B-cell lymphomas including, for example, Hodgkin's lymphoma.
  • T-cell lymphomas including, for example, cutaneous T-cell lymphomas.
  • Hematological malignancies also include leukemias such as secondary leukemia or acute lymphoblastic leukemia.
  • myeloma eg, multiple myeloma
  • other hematological and/or B cell or T cell related cancers include myeloma (eg, multiple myeloma) and other hematological and/or B cell or T cell related cancers.
  • Figures 7A-7B Anti-human ROR1 candidate antibody endocytic activity assay (acid wash method)
  • Figure 8 Anti-human ROR1 candidate antibody endocytic activity assay (pHrodo dye (Thermo) method)
  • the molecular biology experimental methods and immunoassay methods used in the present invention basically refer to J. Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, and The method described in F.M. Ausubel et al., Refined Molecular Biology Experiment Guide, 3rd Edition, John Wiley & Sons, Inc., 1995 was performed.
  • Those skilled in the art appreciate that the examples describe the invention by way of example and are not intended to limit the scope of the invention as claimed.
  • the human ROR1 ECD protein with the various tags described above, pcDNA3.4 plasmid encoding the full-length ROR1 sequence, and Ba/F3-ROR1 cells were used to immunize wild-type mice.
  • the following four mouse strains were used for immunization: Balb/c, C57Bl/6, NZB and A/J.
  • Three immunization protocols were used, outlined as follows: 1) First, 1-2 tail vein injections with 100 ⁇ g pcDNA3.4 plasmid containing the complete coding sequence of human ROR1 were used, followed by boosting immunization with 2-4x10 6 Ba/F3-ROR1 cells, each The immunization interval is 2-3 weeks; 2) First, use 100 ⁇ g of pcDNA3.4 plasmid containing the complete coding sequence of human ROR1 to inject 1-2 times into the tail vein, and then mix 50 ⁇ -100 ⁇ g of recombinant ROR1 with incomplete Freund's adjuvant (IFA).
  • IFA incomplete Freund's adjuvant
  • ECD protein enhances immunization, and each immunization interval is 2-3 weeks; 3) First, 50 ⁇ g recombinant ROR1 ECD protein and complete Freund's adjuvant (CFA) were mixed for immunization, and then 50 ⁇ g recombinant ROR1 ECD protein and IFA were mixed for enhanced immunization. The immunization interval is 2-3 weeks. From the second boost, serum titers were measured by flow cytometry using Ba/F3-ROR1 cells. Mice with high titers were selected for booster immunization with ROR-1 ECD protein 3-5 days before fusion.
  • CFA complete Freund's adjuvant
  • Mouse spleen cells and mouse myeloma SP2/0 cells were fused using standard chemical fusion procedures using polyethylene glycol (molecular weight 1500 Da, Roche, Cat. No. 10 783 641 001). Together, the ratio is 5:1.
  • Cells were adjusted to 5 x 105 splenocytes/mL with DMEM medium (Gibco, Product No. 12430-47) containing 20% fetal bovine serum (Hyclone, Cat. No. SH30080.03). Add 0.2 mL of cells to each well of a 96-well plate and culture in an incubator at 37 °C, 5% CO .
  • HAT Sigma, Cat. No.
  • H0262-10VL was added to each well the day after fusion for screening selection.
  • Clones producing antibodies that bind to Ba/F3-ROR1 cells were selected by high-throughput flow cytometry (Sartorius, model: iQue Screener Plus). Positive clones that bound only to Ba/F3-ROR1 cells but not to Ba/F3 wild-type cells were further selected by flow cytometry (Luminex, model: Guava easyCyte HT). Finally, monoclonal hybridomas were obtained by limiting dilution and their binding to ROR1 cells was confirmed by FACS. Small-scale antibody purification was performed from each selected positive monoclonal hybridoma.
  • Hybridoma cells were cultured in 50-100 mL of serum-free medium (Thermo Fisher, Hybridoma-SFM, Cat. No. 12045084), and the antibody in the supernatant was purified by Protein A (GE Healthcare, MabSelect SuRe, Cat. No. 17543802) and eluted, The eluted monoclonal antibody was dialyzed against 150 mM NaCl. The dialyzed antibody was filtered and filter sterilized through a 0.2 ⁇ m filter.
  • a mouse IgG isotype control (Thermo Fisher, catalog number: 31903) was also used as a negative control.
  • Ba/F3-ROR1, Jeko-1, HT-29 and A549 cells were incubated with different concentrations of purified murine antibody and control antibody on ice for 30 minutes, then washed with flow buffer (PBS+2% FBS) twice, and then incubated with goat anti-human or mouse fluorescent secondary antibody (Jackson ImmunoResearch, product number: 109-605-098 or 115-605-071) on ice for 30 minutes, and finally detected on the machine (Luminex, model: Guava easyCyte HT) ).
  • the candidate antibodies were confirmed to have higher binding specificity to human ROR1 than human ROR2 by ELISA using the extracellular domain (ECD) protein of human ROR2.
  • ECD extracellular domain
  • the specific method is as follows: 1 ⁇ g/mL human ROR2 extracellular domain (ECD) protein (Sino Biologicals, Item No.: 16133-H08H) 100 ⁇ L/well, and coat 96-well ELISA plate (Thermo Fisher, Item No.: 5129) overnight at 4°C.
  • washing buffer (1 ⁇ TBS containing 0.05% Tween-20)
  • MAB2064 mouse anti-human ROR2 Antibody
  • Example 3 Subtype identification and variable region amplification of anti-human ROR1 murine antibody
  • variable region gene total RNA was extracted from the lysed hybridoma cells, and the first-strand cDNA was synthesized using a cDNA reverse transcription kit (purchased from Thermo Fisher, Cat. No. 18080-200), and the PCR method was performed using special antibody primers.
  • VH and VK genes were amplified using Platinum Taq DNA polymerase (purchased from Thermo Fisher, Cat. No. 1304011), and the PCR products were purified by DNA purification kit (purchased from Qiagen, Cat. No. 28104) and ligated into TOPO TA cloning vector (purchased from Thermo Fisher, Cat. No. K457540) for E.coli transformation. Approximately 6-12 E.
  • the binding affinity of purified anti-human ROR1 chimeric antibodies to Ba/F3-ROR1, Jeko-1, HT-29 and A549 cells was analyzed by flow cytometry.
  • the specific experimental steps are as follows: Ba/F3-ROR1, Jeko-1, HT-29 and A549 cells were incubated with different concentration gradients of purified chimeric antibodies on ice for 30 minutes, and the anti-ROR1 antibody UC961 with human IgG1 was used respectively. and anti-TNP antibody as positive and negative controls. Washed twice with flow buffer (PBS+2% FBS), then incubated with goat anti-human fluorescent secondary antibody (Jackson ImmunoResearch, Cat. No.: 109-605-098) on ice for 30 minutes, and finally used flow cytometry detection.
  • Octet ForteBio is widely used in the detection of dynamic affinity of antibody antigens.
  • Candidate chimeric antibodies 3D8-Chi, 19F6-Chi and 38F8-Chi and positive control antibody UC961 use this method to determine the dynamic affinity of human ROR1.
  • the anti-human IgG Fc AHC probe (ForteBio, product number: 18-5060) is first combined with the antibody to be tested to a response signal value of 0.8-1.0 nm, and the antibody-coated probe is immersed in different concentrations (4, 2, 1, 0.5, 0.25, 0.13, 0.06, and 0 ⁇ g/mL) of human ROR1 ECD protein in wells to measure dynamic affinity, binding for 5 minutes, followed by dissociation for 10 minutes, a 1:1 kinetic binding model was used for all simulations. Combined analysis.
  • the dissociation rate of 3D8-Chi, 19F6-Chi and 38F8-Chi was significantly slower than that of the control antibody UC961 (as shown by the Kdis value), and the affinity of 3D8-Chi, 19F6-Chi and 38F8-Chi was higher than that of the control antibody UC961 was 2.0, 34.6 and 5.0 times stronger, respectively (as indicated by KD values).
  • the antibody of the present invention showed stronger antigen-binding ability.
  • humanized antibodies are produced by replacing most or all of the framework sequences of a murine antibody with the corresponding human germline antibody framework sequences, and retain the antigen-specific binding variable regions or CDRs of the murine antibody.
  • hybrid molecules are created in which only the CDRs consist of non-human sequences.
  • Back-mutations typically graft some key murine amino acids into the framework of a human germline antibody for potential deamidation, isomerization, and glycosylation sites in the CDRs in order to maximize retention of the antigen-binding affinity of the parent antibody Mutations were made to improve biophysical properties.
  • the coding DNA sequences of the candidate antibodies (3D8, 38F8, 19F6) in Example 5 were synthesized and codon-optimized, and then cloned into the pcDNA3.4 plasmid (entrusted by Suzhou Hongxun). Technology Co., Ltd.).
  • the pcDNA3.4 plasmids corresponding to the heavy and light chains of each humanized antibody were simultaneously transfected into Expi293F cells, and protein A was used to purify the expressed antibody in the supernatant.
  • the affinity of anti-human ROR1 antibody to Ba/F3-ROR1, Jeko-1, HT-29 and A549 cells was detected by flow cytometry.
  • the specific experimental steps are as follows: Ba/F3-ROR1, Jeko-1, HT-29 and A549 cells were incubated with different concentration gradients of purified humanized antibodies on ice for 30 minutes, and anti-ROR1 antibodies with human IgG1 were used respectively.
  • UC961 and anti-TNP antibody served as positive and negative controls. Washed twice with flow buffer (PBS+2% FBS), then incubated with goat anti-human fluorescent secondary antibody (Jackson ImmunoResearch, Cat. No.: 109-605-098) on ice for 30 minutes, and finally detected on the machine.
  • the humanized molecules 19F6-Hu35V1, 3D8-HuC24 and 38F8-Hu57 and the positive control antibody UC961 were assayed for dynamic affinity with the human ROR1 ECD protein using the Octet ForteBio method.
  • the specific experimental steps are as follows: the anti-human IgG Fc AHC probe (ForteBio, product number: 18-5060) is first combined with the antibody to be tested to a response signal value of 0.8-1.2 nm, and the antibody-coated probe is immersed in different concentrations (16, 8, 4, 2, 1, 0.5, 0.25, and 0 ⁇ g/mL) of human ROR1 ECD protein in wells to measure dynamic affinity, binding for 2-4 minutes followed by dissociation for 4-6 minutes, 1:1 kinetic binding model for all fit analyses.
  • the dissociation rate of the humanized molecule 19F6-Hu35V1 was significantly slower than that of the control antibody UC961 (as shown by the Kdis value), and the affinity was 5.4 times stronger than that of the control antibody UC961 (as shown by the KD value).
  • 3D8-HuC24 and 38F8-Hu57 showed comparable affinity to the control antibody UC961.
  • the humanized anti-human ROR1 candidate antibodies 19F6-Hu35V1, 3D8-HuC24 and 38F8-Hu57 are IgG1 subtypes with strong antibody-dependent cell-mediated cytotoxicity (ADCC) activity .
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • the present invention adopts the method of killing NK cells for detection.
  • the specific experimental steps are as follows: take Ba/F3-ROR1 cells, resuspend them in medium after centrifugation, adjust the cell density to 1 ⁇ 10 4 cells/40 ⁇ L/well and add them to the corresponding wells of a 96-well plate; take NK92MI-mCD16a-hFceRI After the cells were pelleted by centrifugation, they were resuspended in empty MEMA medium, and the cell density was adjusted to 5 ⁇ 10 4 /50 ⁇ L/well and added to the corresponding wells; 19F6-Hu35V1, 3D8-HuC24, 38F8-Hu57 and UC961 were diluted with empty MEMA medium. Antibody, 1000nM starting, 4-fold dilution, 10 concentration points.
  • Antibody binding to cell surface target proteins may induce internalization of antibody-antigen complexes.
  • Mab-ZAP kit (ATSBio, IT-04) was used to measure the inducing endocytosis activity of mouse anti-human ROR1 candidate antibody (Kohls MD and Lappi DA BioTechniques 2000, 28:162). The specific experimental steps are as follows: use a cell counter to count Jeko-1, adjust the cell concentration to 7500/well, then add 90uL of cells to a 96-well plate, and incubate overnight in a 37°C, 5% CO2 incubator.
  • mice antibodies 19F6.2.3, 3D8.2.3, 38F8.1.1, and UC961 control antibodies were diluted 5 times with complete medium from 10ug/ml, with 8 concentration points. 10ul of different concentrations of antibodies were transferred into Jeko-1 cells. Prepare a Mab-zap solution with a concentration of 10ug/mL (50 ⁇ ) according to the kit instructions, add 2ul to each well, and incubate the cells for 3 days at 37°C, 5% CO 2 . Pre-warm CTG2.0 (Promega, Cat. No.
  • the invention adopts the acid washing method to measure the endocytosis activity of the anti-human ROR1 candidate antibody.
  • the specific experimental steps are as follows: after trypsinization of HCC827 and MDA-MB-231 cells, the cell concentration was adjusted to 200,000 cells/well after counting, washed twice with PBS, and resuspended with 2% BSA. added to a 96-well plate.
  • the antibodies 19F6-Hu35V1, 38F8-Hu57, UC961 control antibody, and IgG antibody (chicken lysozyme antibody) were diluted 3 times from 15 ⁇ g/mL, with 8 or 9 concentration points.
  • Plate 1 was placed at 37 °C, and plate 2 was placed at 4 °C for further incubation for 4 h. After the incubation, the medium was removed by centrifugation at 500g for 3min, and 100 ⁇ L of acid (150mM NaCl, 0.1M glycine, adjusted to pH 2.0 with hydrochloric acid) was added to each well, resuspended and washed for 2 minutes, and 60ul of alkali (3g/L Tris base, 9g/L NaCl, adjust the pH to 13 with NaOH), mix and neutralize, remove the supernatant, add PBS to resuspend, and then detect by flow meter.
  • acid 150mM NaCl, 0.1M glycine, adjusted to pH 2.0 with hydrochloric acid
  • the present invention simultaneously adopts pHrodo dye (Thermo) to measure the endocytosis activity of the anti-human ROR1 candidate antibody.
  • the specific experimental steps are as follows: trypsinize N87 cells, use a cell counter to count the cells, adjust the cell concentration to 10,000 cells/well, take 100 ⁇ L of cells and spread them in a 96-well plate, and incubate in an incubator at 37°C, 5% CO 2 . overnight.
  • the antibody 19F6-Hu35V1, UC961 control antibody, and IgG antibody were diluted 5 times with 1640 complete medium starting from 2.4 ⁇ g/ml, with 7 concentration points.
  • pHrodo TM Red dye was diluted with 1640 complete medium to 12 ⁇ g/mL, 30 ⁇ L of different concentrations of antibodies were mixed with 30 ⁇ L of the diluted dye, and incubated at room temperature for 20 min in the dark. After the incubation, the cell culture medium was discarded, the antibody-dye mixture was added to each well of cells, and the cells were incubated overnight at 37°C in a 5% CO 2 incubator. The next day, cells were digested with trypsin and detected by flow cytometry. The results are shown in FIG. 8 , the endocytosis activity of the antibody 19F6-Hu35V1 in N87 cells was stronger than that of the control antibody UC961, about 7 times that of UC961. The negative control antibody IgG has no endocytic activity in N87.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Cell Biology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
PCT/CN2022/078327 2021-03-09 2022-02-28 Ror1结合蛋白及其用途 Ceased WO2022188652A1 (zh)

Priority Applications (7)

Application Number Priority Date Filing Date Title
CA3207635A CA3207635A1 (en) 2021-03-09 2022-02-28 Ror1 binding protein and use thereof
JP2023541591A JP2024508597A (ja) 2021-03-09 2022-02-28 Ｒｏｒ１結合タンパク質及びその用途
US18/260,556 US20240018237A1 (en) 2021-03-09 2022-02-28 Ror1 binding protein and use thereof
EP22766178.2A EP4306540A4 (en) 2021-03-09 2022-02-28 ROR1-BINDING PROTEIN AND USE THEREOF
CN202280009364.9A CN116829599A (zh) 2021-03-09 2022-02-28 Ror1结合蛋白及其用途
MX2023008176A MX2023008176A (es) 2021-03-09 2022-02-28 Proteina de union a ror1 y uso de la misma.
KR1020237024374A KR20230156021A (ko) 2021-03-09 2022-02-28 Ror1 결합 단백질 및 이의 용도

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202110255430.8 2021-03-09
CN202110255430 2021-03-09

Publications (1)

Publication Number Publication Date
WO2022188652A1 true WO2022188652A1 (zh) 2022-09-15

Family

ID=83226356

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/078327 Ceased WO2022188652A1 (zh) 2021-03-09 2022-02-28 Ror1结合蛋白及其用途

Country Status (8)

Country Link
US (1) US20240018237A1 (https=)
EP (1) EP4306540A4 (https=)
JP (1) JP2024508597A (https=)
KR (1) KR20230156021A (https=)
CN (1) CN116829599A (https=)
CA (1) CA3207635A1 (https=)
MX (1) MX2023008176A (https=)
WO (1) WO2022188652A1 (https=)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2022227686A1 (en) 2021-02-25 2023-07-27 Lyell Immunopharma, Inc. Ror1 targeting chimeric antigen receptor

Citations (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3817837A (en) 1971-05-14 1974-06-18 Syva Corp Enzyme amplification assay
US3850752A (en) 1970-11-10 1974-11-26 Akzona Inc Process for the demonstration and determination of low molecular compounds and of proteins capable of binding these compounds specifically
US3939350A (en) 1974-04-29 1976-02-17 Board Of Trustees Of The Leland Stanford Junior University Fluorescent immunoassay employing total reflection for activation
US3996345A (en) 1974-08-12 1976-12-07 Syva Company Fluorescence quenching with immunological pairs in immunoassays
US4275149A (en) 1978-11-24 1981-06-23 Syva Company Macromolecular environment control in specific receptor assays
US4277437A (en) 1978-04-05 1981-07-07 Syva Company Kit for carrying out chemically induced fluorescence immunoassay
US4366241A (en) 1980-08-07 1982-12-28 Syva Company Concentrating zone method in heterogeneous immunoassays
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
WO2002043478A2 (en) 2000-11-30 2002-06-06 Medarex, Inc. Transgenic transchromosomal rodents for making human antibodies
CN104341529A (zh) 2013-07-26 2015-02-11 三星电子株式会社 带有DARPin的双特异性嵌合蛋白
WO2015141862A1 (en) 2014-03-17 2015-09-24 Mitsubishi Tanabe Pharma Corporation Antibody-fynomer conjugates
US9217040B2 (en) 2011-01-14 2015-12-22 The Regents Of The University Of California Therapeutic antibodies against ROR-1 protein and methods for use of same
CN105924533A (zh) * 2016-07-13 2016-09-07 北京康爱瑞浩生物科技股份有限公司 Ror1特异性嵌合抗原受体及其应用
CN108441505A (zh) * 2018-05-28 2018-08-24 上海恒润达生生物科技有限公司 一种靶向ror1的嵌合抗原受体及其用途
WO2018237335A1 (en) 2017-06-23 2018-12-27 VelosBio Inc. IMMUNOCONJUGUATED ROR1 ANTIBODIES
CN109836498A (zh) * 2017-11-25 2019-06-04 深圳宾德生物技术有限公司 一种靶向ror1的单链抗体、嵌合抗原受体t细胞及其制备方法和应用
WO2020014366A1 (en) * 2018-07-10 2020-01-16 Intrexon Corporation Ror-1 specific chimeric antigen receptors and uses thereof
WO2020160050A1 (en) * 2019-01-29 2020-08-06 Juno Therapeutics, Inc. Antibodies and chimeric antigen receptors specific for receptor tyrosine kinase like orphan receptor 1 (ror1)

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9758586B2 (en) * 2010-12-01 2017-09-12 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Chimeric rabbit/human ROR1 antibodies
GB201721802D0 (en) * 2017-12-22 2018-02-07 Almac Discovery Ltd Ror1-specific antigen binding molecules

Patent Citations (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3850752A (en) 1970-11-10 1974-11-26 Akzona Inc Process for the demonstration and determination of low molecular compounds and of proteins capable of binding these compounds specifically
US3817837A (en) 1971-05-14 1974-06-18 Syva Corp Enzyme amplification assay
US3939350A (en) 1974-04-29 1976-02-17 Board Of Trustees Of The Leland Stanford Junior University Fluorescent immunoassay employing total reflection for activation
US3996345A (en) 1974-08-12 1976-12-07 Syva Company Fluorescence quenching with immunological pairs in immunoassays
US4277437A (en) 1978-04-05 1981-07-07 Syva Company Kit for carrying out chemically induced fluorescence immunoassay
US4275149A (en) 1978-11-24 1981-06-23 Syva Company Macromolecular environment control in specific receptor assays
US4366241A (en) 1980-08-07 1982-12-28 Syva Company Concentrating zone method in heterogeneous immunoassays
US4366241B1 (https=) 1980-08-07 1988-10-18
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
US5693762A (en) 1988-12-28 1997-12-02 Protein Design Labs, Inc. Humanized immunoglobulins
US5585089A (en) 1988-12-28 1996-12-17 Protein Design Labs, Inc. Humanized immunoglobulins
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
US6180370B1 (en) 1988-12-28 2001-01-30 Protein Design Labs, Inc. Humanized immunoglobulins and methods of making the same
WO2002043478A2 (en) 2000-11-30 2002-06-06 Medarex, Inc. Transgenic transchromosomal rodents for making human antibodies
US9217040B2 (en) 2011-01-14 2015-12-22 The Regents Of The University Of California Therapeutic antibodies against ROR-1 protein and methods for use of same
CN104341529A (zh) 2013-07-26 2015-02-11 三星电子株式会社 带有DARPin的双特异性嵌合蛋白
WO2015141862A1 (en) 2014-03-17 2015-09-24 Mitsubishi Tanabe Pharma Corporation Antibody-fynomer conjugates
CN105924533A (zh) * 2016-07-13 2016-09-07 北京康爱瑞浩生物科技股份有限公司 Ror1特异性嵌合抗原受体及其应用
WO2018237335A1 (en) 2017-06-23 2018-12-27 VelosBio Inc. IMMUNOCONJUGUATED ROR1 ANTIBODIES
CN109836498A (zh) * 2017-11-25 2019-06-04 深圳宾德生物技术有限公司 一种靶向ror1的单链抗体、嵌合抗原受体t细胞及其制备方法和应用
CN108441505A (zh) * 2018-05-28 2018-08-24 上海恒润达生生物科技有限公司 一种靶向ror1的嵌合抗原受体及其用途
WO2020014366A1 (en) * 2018-07-10 2020-01-16 Intrexon Corporation Ror-1 specific chimeric antigen receptors and uses thereof
WO2020160050A1 (en) * 2019-01-29 2020-08-06 Juno Therapeutics, Inc. Antibodies and chimeric antigen receptors specific for receptor tyrosine kinase like orphan receptor 1 (ror1)

Non-Patent Citations (51)

* Cited by examiner, † Cited by third party
Title
"Antibody Engineering: Methods and Protocols", vol. 248, 2004, HUMANA PRESS
ALFTHAN ET AL., PROTEIN ENG., vol. 8, 1995, pages 725 - 731
ALTSCHUL SF ET AL., NATURE GENET., vol. 6, 1994, pages 119
BIRD ET AL., SCIENCE, vol. 242, 1988, pages 423 - 426
BRENNAN ET AL., SCIENCE, vol. 229, 1985, pages 81
BRODEUR ET AL.: "Monoclonal Antibody Production Techniques and Applications", 1987, MARCEL DEKKER, INC., pages: 51 - 63
BRUGGERMANN ET AL., YEAR IN IMMUNOLOGY, vol. 7, 1993, pages 33
BRUMMELL ET AL., BIOCHEM., vol. 32, 1993, pages 1180 - 1187
BURKS ET AL., PROC. NATL ACAD. SET USA, vol. 94, 1997, pages 412 - 417
CARTER ET AL., BIO/TECHNOLOGY, vol. 10, 1992, pages 163 - 167
CHOI ET AL., EUR. J. IMMUNOL., vol. 31, 2001, pages 94 - 106
CHOTHIA ET AL., NATURE, vol. 341, 1989, pages 544 - 546
CHOTHIALESK, J. MOL. BIOL., vol. 196, 1987, pages 901 - 917
CLACKSON ET AL., NATURE, vol. 349, 1991, pages 293 - 299
D. WILKINSON: "The Scientist", vol. 14, 17 April 2000, THE SCIENTIST, INC., pages: 25 - 28
DAVIES ET AL., ANNUAL REV BIOCHEM, vol. 59, 1990, pages 439 - 473
DUCHOSAL ET AL., NATURE, vol. 355, 1992, pages 258
E. MEYERSW. MILLER, COMPUT. APPL. BIOSCI., vol. 4, 1988, pages 11 - 17
F. M. AUSUBEL ET AL.: "Short Protocols in Molecular Biology", 1995, JOHN WILEY & SONS, INC.
HIGGINS DGSHARP PM, CABIOS, vol. 5, 1989, pages 151
HOLLIGER ET AL., NAT BIOTECHNOL, vol. 23, 2005, pages 1126 - 1136
HOOGENBOOM ET AL., J. MOL. BIOL., vol. 227, 1991, pages 381 - 597
HU ET AL., CANCER RES., vol. 56, 1996, pages 3055 - 3061
HUDSON, CURR. OPIN. IMMUNOL., vol. 11, 1999, pages 548 - 557
HUSTON ET AL., PROC. NATL. ACAD. SCI. USA, vol. 85, 1988, pages 5879 - 5883
JAKOBOVITS ET AL., NATURE, vol. 361, 1993, pages 186 - 187
JAKOBOVITS ET AL., PROC. NATL. ACAD. SCI. USA, vol. 90, 1993, pages 2551 - 6448
JONES ET AL., NATURE, vol. 321, 1986, pages 522 - 525
KABAT, E.A. ET AL.: "Sequences of Proteins of Immunological Interest", 1991, U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES
KIPRIYANOV ET AL., J. MOL. BIOL., vol. 293, 1999, pages 41 - 56
KOBAYASHI ET AL., PROTEIN ENG., vol. 12, no. 10, 1999, pages 879 - 884
KOHLER ET AL., NATURE, vol. 256, 1975, pages 495
KOZBOR, J. IMMUNOL., vol. 133, 1984, pages 3001
LEFRANC ET AL., DEV. COMPARAT. IMMUNOL., vol. 27, 2003, pages 55 - 77
LITTLE ET AL., IMMUNOL. TODAY, vol. 21, 2000, pages 397 - 402
LONBERG ET AL., NATURE, vol. 368, no. 6474, 1994, pages 856 - 859
MARTIN ACRCHEETHAM JCREES AR: "Modeling antibody hypervariable loops: A combined algorithm", PROC NATL ACAD SCI USA, vol. 86, 1989, pages 9268 - 9272, XP000165667, DOI: 10.1073/pnas.86.23.9268
MORIMOTO ET AL., J. BIOCHEM. BIOPHYS. METHODS, vol. 24, 1992, pages 107 - 117
MORRISON ET AL., PROC. NATL. ACAD. SCI. USA, vol. 81, 1984, pages 6851 - 6855
MUNSON ET AL., ANAL. BIOCHEM., vol. 107, 1980, pages 220
NEEDLEMAN ET AL., J. MOL. BIOL., vol. 48, 1970, pages 444 - 453
POLJAK R. J. ET AL., STRUCTURE, vol. 113, 1994, pages 1121 - 1123
PRESTA, CURR. OP. STRUCT. BIOL., vol. 2, 1992, pages 593 - 596
QUEEN ET AL., PROC. NATL. ACAD. SCI. USA, vol. 86, 1989, pages 10029 - 10033
RADER ET AL., PROC. NAT. ACAD. SCI. USA, vol. 95, 1998, pages 8910 - 8915
REICHMANN ET AL., NATURE, vol. 332, 1988, pages 323 - 329
ROOVERS ET AL., CANCER IMMUNOL, 2001
See also references of EP4306540A4
SMITH TFWATERMAN MS, ADV. APPL. MATH., vol. 2, 1981, pages 482
STEINBERGER ET AL., J. BIOL. CHEM., vol. 275, 2000, pages 36073 - 36078
VAUGHAN ET AL., NATURE BIOTECH, vol. 14, 1996, pages 309 - 103

Also Published As

Publication number Publication date
KR20230156021A (ko) 2023-11-13
CN116829599A (zh) 2023-09-29
EP4306540A4 (en) 2025-05-28
EP4306540A1 (en) 2024-01-17
CA3207635A1 (en) 2022-09-15
JP2024508597A (ja) 2024-02-28
US20240018237A1 (en) 2024-01-18
MX2023008176A (es) 2023-07-18

Similar Documents

Publication Publication Date Title
US20250340630A1 (en) Antibody and use thereof
CN115043942A (zh) Ror1结合蛋白及其用途
US20240425593A1 (en) Anti-pd-1 antibody and use thereof
EP4596582A1 (en) B7-h3 binding protein and use thereof
CA3259098A1 (en) ANTIBODIES AND THEIR USES
WO2022188652A1 (zh) Ror1结合蛋白及其用途
EP4488294A1 (en) Her3 binding protein and use thereof
WO2024007671A1 (zh) 特异性结合cd24的抗体及其用途
CN117801106A (zh) 抗pd-1抗体及其用途
CN117586398A (zh) 抗tigit抗体及其用途
CN117794952A (zh) 抗tigit抗体及其用途
WO2025256584A1 (en) Cdh3 binding protein and use thereof
WO2025214479A1 (en) Ceacam5 binding protein and use thereof
WO2025124443A1 (zh) Adam9结合蛋白及其用途
CN121226558A (zh) Ceacam5结合蛋白及其用途
TW202417516A (zh) Ptk7結合蛋白及其用途
HK40048164A (en) Antibody and use thereof
HK40048164B (zh) 一种抗体及其用途

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: 22766178

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 18260556

Country of ref document: US

ENP Entry into the national phase

Ref document number: 3207635

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2023541591

Country of ref document: JP

Ref document number: 202280009364.9

Country of ref document: CN

Ref document number: MX/A/2023/008176

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 2022766178

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2022766178

Country of ref document: EP

Effective date: 20231009