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CN-121986115-A - Anti-PD-1 monoclonal antibody and application thereof

CN121986115ACN 121986115 ACN121986115 ACN 121986115ACN-121986115-A

Abstract

Relates to the technical field of antibody medicaments, in particular to an anti-PD-1 monoclonal antibody and application thereof. An antibody or antigen-binding fragment thereof that specifically binds to PD-1, pharmaceutical compositions containing the antibody or antigen-binding fragment thereof, nucleic acid molecules encoding the antibody or antigen-binding fragment thereof, and host cells comprising the nucleic acid molecules, and therapeutic and diagnostic uses of the antibody or antigen-binding fragment thereof are provided.

Inventors

  • JIANG WENBO
  • WANG SIQIN
  • JIN LEI
  • XU LIZHONG
  • CHU XUEBIN
  • SONG LEI
  • LI XUE
  • XUE WEILI
  • LI LINGYUN
  • ZHAO RANRAN
  • HE XUZHI

Assignees

  • 上海赛增医疗科技有限公司

Dates

Publication Date
20260505
Application Date
20241009
Priority Date
20231010

Claims (20)

  1. An antibody or antigen-binding fragment thereof capable of specifically binding to PD-1, the antibody or antigen-binding fragment thereof comprising: (a) VH CDR1 or variant thereof, VH CDR2 or variant thereof and VH CDR3 or variant thereof contained in the heavy chain variable region (VH) as shown in SEQ ID NO 7 or 15 and/or VL CDR1 or variant thereof, VL CDR2 or variant thereof and VL CDR3 or variant thereof contained in the light chain variable region (VL) as shown in SEQ ID NO 8 or 16 or (B) A VH CDR1 or variant thereof, a VH CDR2 or variant thereof and a VH CDR3 or variant thereof contained in a heavy chain variable region (VH) as shown in SEQ ID NO 31 or 39 and/or a VL CDR1 or variant thereof, a VL CDR2 or variant thereof and a VL CDR3 or variant thereof contained in a light chain variable region (VL) as shown in SEQ ID NO 32 or 40; Wherein the variant has a substitution, deletion or addition of one or more amino acids (e.g., a substitution, deletion or addition of 1,2 or 3 amino acids, e.g., a conservative substitution) compared to the sequence from which it is derived, preferably the substitution is a conservative substitution; preferably, the antibody or antigen binding fragment thereof comprises: (a) 3 CDRs contained in the heavy chain variable region (VH) as shown in SEQ ID NO 7 or 15, and/or 3 CDRs contained in the light chain variable region (VL) as shown in SEQ ID NO 8 or 16, or, (B) 3 CDRs contained in the heavy chain variable region (VH) as shown in SEQ ID NO. 31 or 39, and/or 3 CDRs contained in the light chain variable region (VL) as shown in SEQ ID NO. 32 or 40.
  2. The antibody or antigen-binding fragment thereof of claim 1, comprising: (a) A heavy chain variable region (VH) comprising 3 Complementarity Determining Regions (CDRs) comprising VH CDR1 of SEQ ID NO. 1, VH CDR2 of SEQ ID NO. 2, VH CDR3 of SEQ ID NO. 3, and/or a light chain variable region (VL) comprising 3 Complementarity Determining Regions (CDRs) comprising VL CDR1 of SEQ ID NO. 4, VL CDR2 of SEQ ID NO. 5, VL CDR3 of SEQ ID NO. 6, or, (B) A heavy chain variable region (VH) comprising 3 Complementarity Determining Regions (CDRs) comprising VH CDR1 of SEQ ID NO. 25, VH CDR2 of SEQ ID NO. 26, VH CDR3 of SEQ ID NO. 27, and/or a light chain variable region (VL) comprising 3 Complementarity Determining Regions (CDRs) comprising VL CDR1 of SEQ ID NO. 28, VL CDR2 of SEQ ID NO. 29, VL CDR3 of SEQ ID NO. 30; wherein the CDRs are defined by the Kabat numbering system.
  3. The antibody or antigen-binding fragment thereof of claim 1 or 2, comprising: (a) A heavy chain variable region (VH) comprising the sequence shown in SEQ ID NO. 7 or a variant thereof and/or a light chain variable region (VL) comprising the sequence shown in SEQ ID NO. 8 or a variant thereof or, (B) A heavy chain variable region (VH) comprising the sequence shown in SEQ ID NO. 31 or a variant thereof and/or a light chain variable region (VL) comprising the sequence shown in SEQ ID NO. 32 or a variant thereof; Wherein the variant has a substitution, deletion, or addition of one or more amino acids (e.g., a substitution, deletion, or addition of 1,2,3, 4, or 5 amino acids) or has 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 compared to the sequence from which it was derived; preferably, the antibody or antigen-binding fragment thereof comprises a VH comprising the sequence shown as SEQ ID NO. 7 and a VL comprising the sequence shown as SEQ ID NO. 8; Preferably, the antibody or antigen-binding fragment thereof comprises a VH comprising the sequence shown as SEQ ID NO. 31 and a VL comprising the sequence shown as SEQ ID NO. 32.
  4. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof comprises a framework region sequence derived from a human immunoglobulin; Preferably, the antibody or antigen binding fragment thereof comprises a heavy chain framework region of a human heavy chain germline sequence and/or a light chain framework region of a human light chain germline sequence.
  5. The antibody or antigen-binding fragment thereof of claim 4, wherein the antibody or antigen-binding fragment thereof comprises: (a) A heavy chain variable region (VH) comprising a sequence as set forth in SEQ ID NO. 15 or a variant thereof and/or a light chain variable region (VL) comprising a sequence as set forth in SEQ ID NO. 16 or a variant thereof or, (B) A heavy chain variable region (VH) comprising a sequence as set forth in SEQ ID NO. 39 or a variant thereof and/or a light chain variable region (VL) comprising a sequence as set forth in SEQ ID NO. 40 or a variant thereof; Wherein the variant has a substitution, deletion, or addition of one or more amino acids (e.g., a substitution, deletion, or addition of 1,2, 3, 4, or 5 amino acids) as compared to the sequence from which it is derived, or has 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; Preferably, the substitution is a conservative substitution; preferably, the antibody or antigen binding fragment thereof comprises: (a) Comprising the heavy chain variable region (VH) as shown in SEQ ID NO. 15 and/or comprising the light chain variable region (VL) as shown in SEQ ID NO. 16 or, (B) Comprising a heavy chain variable region (VH) as shown in SEQ ID NO 39 and/or comprising a light chain variable region (VL) as shown in SEQ ID NO 40.
  6. The antibody or antigen-binding fragment thereof of any one of claims 1-5, wherein the antibody or antigen-binding fragment thereof further comprises a constant region derived from a mammalian (e.g., human or murine) immunoglobulin; Preferably, the heavy chain of the antibody or antigen binding fragment thereof comprises a heavy chain constant region derived from a human immunoglobulin (e.g., igG, such as IgG1, igG2, igG3 or IgG 4), and/or the light chain of the antibody or antigen binding fragment thereof comprises a light chain constant region derived from a human immunoglobulin (e.g., kappa or lambda).
  7. The antibody or antigen-binding fragment thereof of claim 6, wherein the antibody or antigen-binding fragment thereof comprises a human immunoglobulin heavy chain constant region variant that has enhanced binding activity to an Fc receptor (e.g., CD32A, CD B and/or CD 16A) as compared to a wild-type human immunoglobulin heavy chain constant region; Preferably, the antibody or antigen binding fragment thereof comprises a human IgG1 or IgG4 heavy chain constant region variant comprising the substitutions 239, 268, preferably selected from S239D/H268D or S239D/Q268D, at one or more of the following positions according to EU numbering.
  8. The antibody or antigen-binding fragment thereof of claim 6 or 7, wherein the antibody or antigen-binding fragment thereof comprises: (1) A heavy chain comprising the sequence shown in SEQ ID NO. 11 and a light chain comprising the sequence shown in SEQ ID NO. 12; (2) A heavy chain comprising the sequence shown in SEQ ID NO. 19 and a light chain comprising the sequence shown in SEQ ID NO. 20; (3) A heavy chain comprising the sequence shown in SEQ ID NO. 23 and a light chain comprising the sequence shown in SEQ ID NO. 20; (4) A heavy chain comprising the sequence shown in SEQ ID NO.35 and a light chain comprising the sequence shown in SEQ ID NO. 36; (5) A heavy chain comprising the sequence shown in SEQ ID NO. 43 and a light chain comprising the sequence shown in SEQ ID NO. 44, or, (6) A heavy chain comprising the sequence shown in SEQ ID NO. 47 and a light chain comprising the sequence shown in SEQ ID NO. 44.
  9. The antibody or antigen-binding fragment thereof of any one of claims 1-8, wherein the antigen-binding fragment is selected from the group consisting of Fab, fab ', F (ab') 2 , fd, fv, disulfide-linked Fv, scFv, di-scFv, (scFv) 2 , diabody, and single domain antibody (sdAb), and/or the antibody is a murine, humanized, chimeric, bispecific, or multispecific antibody.
  10. An isolated nucleic acid molecule encoding the antibody or antigen-binding fragment thereof of any one of claims 1-9, or a heavy chain variable region and/or a light chain variable region thereof, or a heavy chain and/or a light chain thereof.
  11. A vector comprising the isolated nucleic acid molecule of claim 10, preferably the vector is a cloning vector or an expression vector.
  12. A host cell comprising the isolated nucleic acid molecule of claim 10 or the vector of claim 11.
  13. A method of making the antibody or antigen-binding fragment thereof of any one of claims 1-9, comprising culturing the host cell of claim 12 under conditions that allow expression of the antibody or antigen-binding fragment thereof, and recovering the antibody or antigen-binding fragment thereof from the cultured host cell culture.
  14. A bispecific or multispecific molecule comprising the antibody or antigen-binding fragment thereof of any one of claims 1-9; preferably, the bispecific or multispecific molecule specifically binds PD-1, and additionally specifically binds one or more other targets; Preferably, the bispecific or multispecific molecule further comprises at least one molecule (e.g., a second antibody) having a second binding specificity for a second target.
  15. An immunoconjugate comprising the antibody or antigen-binding fragment thereof of any one of claims 1-9, a therapeutic agent linked to the antibody or antigen-binding fragment thereof; preferably, the therapeutic agent is a cytotoxic agent.
  16. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1-9, the isolated nucleic acid molecule of claim 10, the vector of claim 11, the host cell of claim 12, the bispecific or multispecific molecule of claim 14, or the immunoconjugate of claim 15, and a pharmaceutically acceptable carrier and/or excipient; Preferably, the pharmaceutical composition further comprises an additional pharmaceutically active agent; preferably, the additional pharmaceutically active agent is an immunosuppressant; Preferably, the additional pharmaceutically active agent is a medicament for the treatment of autoimmune diseases, allergic diseases, graft versus host diseases, graft rejection or lymphomas.
  17. Use of the antibody or antigen binding fragment thereof of any one of claims 1-9, the isolated nucleic acid molecule of claim 10, the vector of claim 11, the host cell of claim 12, the bispecific or multispecific molecule of claim 14, the immunoconjugate of claim 15, or the pharmaceutical composition of claim 16 in the preparation of a PD-1 signaling pathway activator.
  18. Use of the antibody or antigen binding fragment thereof of any one of claims 1-9, the isolated nucleic acid molecule of claim 10, the vector of claim 11, the host cell of claim 12, the bispecific or multispecific molecule of claim 14, the immunoconjugate of claim 15, or the pharmaceutical composition of claim 16 in the preparation of an immunosuppressant (e.g., an inhibitor of T cell activity).
  19. Use of the antibody or antigen binding fragment thereof of any one of claims 1-9, the isolated nucleic acid molecule of claim 10, the vector of claim 11, the host cell of claim 12, the bispecific or multispecific molecule of claim 14, the immunoconjugate of claim 15, or the pharmaceutical composition of claim 16 in the manufacture of a medicament for preventing and/or treating an autoimmune disease, an allergic disease, a graft-versus-host disease, a graft rejection, or a lymphoma in a subject; Preferably, the autoimmune disease is selected from arthritis, rheumatoid arthritis, psoriatic arthritis, lupus nephritis, systemic lupus erythematosus, psoriasis, vitiligo, alopecia areata, inflammatory bowel disease, ulcerative colitis, crohn's disease, type I diabetes, multiple sclerosis, autoimmune hepatitis, primary biliary cirrhosis, celiac disease, scleroderma, graves' disease, hashimoto's thyroiditis, ankylosing spondylitis, myasthenia gravis, sjogren's syndrome, igA nephropathy, igG4-related diseases (IgG 4-RELATED DISEASE), vasculitis, ANCA-related vasculitis, uveitis, pemphigus, pemphigoid or autoimmune hemolytic anemia; Preferably, the allergic disease is selected from idiopathic thrombocytopenic purpura, atopic dermatitis, asthma, allergic rhinitis, drug allergies, food allergies, allergic conjunctivitis, urticaria, eosinophilic sinusitis, eosinophilic digestive organ disease or allergic bronchopulmonary aspergillosis; preferably, the lymphoma is a T-cell lymphoma; Preferably, the subject is a mammal, such as a human or a non-human primate (e.g., cynomolgus monkey); preferably, the antibody or antigen binding fragment thereof is used alone or in combination with an additional pharmaceutically active agent, preferably the additional pharmaceutically active agent is an immunosuppressant, preferably the additional pharmaceutically active agent is a medicament for the treatment of autoimmune diseases, allergic diseases, graft versus host diseases, graft rejection or lymphomas.
  20. A conjugate comprising the antibody or antigen-binding fragment thereof of any one of claims 1-9, and a detectable label attached to the antibody or antigen-binding fragment thereof; Preferably, the detectable label is selected from the group consisting of enzymes (e.g., horseradish peroxidase or alkaline phosphatase), chemiluminescent reagents (e.g., acridine esters, luminol and derivatives thereof, or ruthenium derivatives), fluorescent dyes (e.g., fluorescein or fluorescent protein), radionuclides, or biotin.

Description

Anti-PD-1 monoclonal antibody and application thereof Cross Reference to Related Applications The present application claims priority from chinese patent application 202311307741.X filed 10 at 2023 and chinese patent application 202410131767.1 filed 30 at 1 at 2024, the entire contents of which are incorporated herein by reference. Technical Field The invention relates to the technical field of antibody medicines, in particular to an anti-PD-1 monoclonal antibody and application thereof. Technical Field Programmed death-1 (also known as CD 279) is an inhibitory receptor that is expressed predominantly on the surface of activated T cells. B cells, monocytes and dendritic cells of certain subpopulations will also express PD-1 after activation. PD-1 belongs to a member of the immunoglobulin superfamily, having a total length of 288 amino acids, comprising an extracellular domain, a transmembrane domain and a cytoplasmic domain. The extracellular domain contains an immunoglobulin-like domain (IgV-like domain) responsible for binding to its ligand. The cytoplasmic domain contains an immunoreceptor tyrosine repression motif (ITIM) and an immunoreceptor tyrosine transduction motif (ITSM), responsible for signal transduction. PD-1 has two ligands, programmed ligand-1 (Programmed DEATH LIGAND, PD-L1) and Programmed ligand-2 (Programmed DEATH LIGAND, PD-L2), respectively. The initial T cell surface does not express PD-1. When the TCR/CD3 complex and CD28 on the surface of the naive T cell bind to MHC-antigen peptide and CD80/CD86, respectively, the intracellular domains of the TCR/CD3 complex and CD28 are phosphorylated by protein kinases, respectively, thereby recruiting a series of signal transduction proteins and initiating a series of phosphorylation events, ultimately resulting in the naive T cell being activated, and then its surface begins to up-regulate PD-1. When PD-1 binds to PD-L1 or PD-L2 on the surface of antigen presenting cells, PD-1 is activated and its cytoplasmic domain is phosphorylated by protein kinases, thereby recruiting protein phosphatase SHP-2.SHP-2 can dephosphorylate the CD28 and TCR/CD3 complex intracellular, thereby inhibiting T cell activation. As an inhibitory receptor, PD-1 plays an important role in the inhibition of immune responses and in the maintenance of peripheral tolerance. PD-1 gene deficiency can lead to the development of autoimmune diseases. PD-1 knockout C57BL/6 mice spontaneously develop lupus-like glomerulonephritis and arthritic symptoms (H Nishimura et al, immunity 1999Aug;11 (2): 141-51.). NZB/W F1 mice spontaneously develop symptoms similar to human systemic lupus erythematosus, the most commonly used mouse model for human systemic lupus erythematosus study. Treatment with the mouse PD-L1 fusion protein can alleviate the proteinuria symptoms of NZB/W F1 mice and prolong the survival time of the mice (Wenjun Liao et al, am J Nephrol.2017;46 (5): 371-379.). Chicken type two collagen can induce mice to develop arthritic symptoms. Collagen-induced murine models of arthritis are the most common disease model for human rheumatoid arthritis. Treatment with the mouse PD-L1 fusion protein can alleviate the arthritic symptoms in C57BL/6 mice (Amalia P Raptopoulou et al, arthritis Rheum.2010Jul;62 (7): 1870-80.). While there are a number of PD-1 antagonistic antibodies (such as palbociclib, also known as corydalid) that block the PD-1 signaling pathway currently on the market and are used to treat a variety of tumors, it remains necessary to develop PD-1 agonist antibodies that activate the PD-1 signaling pathway for inhibiting T cell responses to treat inflammatory and autoimmune diseases. Disclosure of Invention The inventors of the present application have obtained, through a great deal of research, a murine antibody that specifically binds to PD-1, which has a remarkable effect of activating the PD-1 signaling pathway, and further obtained a humanized antibody that has PD-1 binding activity and PD-1 signaling pathway activation based on the murine antibody. The antibody provided by the application can effectively inhibit T cell activity and immune response or inflammatory response by activating PD-1 signal channels, thereby playing a role in preventing and treating inflammation, allergy and autoimmune related diseases. In addition, the antibody of the application has cross-binding activity and activation activity on the species of human and monkey PD-1, thereby being beneficial to the development of preclinical evaluation research and other works in animal models. Antibodies or antigen binding fragments thereof In a first aspect, the present application provides an antibody or antigen-binding fragment thereof capable of specifically binding to PD-1, the antibody or antigen-binding fragment thereof comprising: A VH CDR1 or variant thereof, a VH CDR2 or variant thereof and a VH CDR3 or variant thereof contained in a heavy chain variable region (VH) as shown in SEQ ID NO 7 or