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RU-2861719-C2 - ANTI-PD-1 ANTIBODIES AND FUSION PROTEINS

RU2861719C2RU 2861719 C2RU2861719 C2RU 2861719C2RU-2861719-C2

Abstract

FIELD: biotechnology. SUBSTANCE: provided are recombinant anti-PD-1 antibodies, antigen-binding fragments thereof and fusion proteins based thereon, as well as nucleic acid molecules encoding same. In addition, provided are therapeutic compositions based on the antibodies and fusion proteins according to the invention and methods of using such antibodies. EFFECT: invention may find further use in therapy for enhancing T-cell and NK-cell function to enhance cellular and cytokine-mediated immunity, as well as for treating cancer and infectious diseases. 42 cl, 118 dwg, 28 tbl, 38 ex

Inventors

  • LU, DAN
  • POLONSKAYA, ZHANNA
  • CHANG, Tzu-Pei
  • MARTOMO, Stella, A.
  • PATEL, Jeegar
  • Miyara, Faical

Dates

Publication Date
20260508
Application Date
20210623
Priority Date
20200623

Claims (20)

  1. 1. An antibody to PD-1 or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein each of the heavy chain and light chain variable regions comprises CDR1, CDR2 and CDR3 and wherein:
  2. the CDR1 sequence of the variable region of the heavy chain (CDR1H) comprises SEQ ID NO: 21;
  3. the heavy chain variable region CDR2 sequence (CDR2H) comprises SEQ ID NO: 22;
  4. the variable region CDR3 sequence of the heavy chain (CDR3H) comprises SEQ ID NO: 23;
  5. the sequence of CDR1, CDR2 and CDR3 of the variable region of the light chain (CDR1L, CDR2L and CDR3L) contains:
  6. i) SEQ ID NOS: 96, 97 and 82;
  7. ii) SEQ ID NOS: 45, 46 and 47;
  8. iii) SEQ ID NOS: 66, 67 and 68;
  9. iv) SEQ ID NOS: 70, 67 and 71;
  10. v) SEQ ID NOS: 73, 74 and 75;
  11. vi) SEQ ID NOS: 77, 78 and 47;
  12. vii) SEQ ID NOS: 80, 81 and 82;
  13. viii) SEQ ID NOS: 77, 78 and 84;
  14. ix) SEQ ID NOS: 77, 86 and 47;
  15. x) SEQ ID NOS: 88, 89 and 47;
  16. xi) SEQ ID NOS: 66, 67 and 47;
  17. xii) SEQ ID NOS: 80, 92 and 75;
  18. xiii) SEQ ID NOS: 80, 94 and 71;
  19. xiv) SEQ ID NOS: 99, 100 and 47;
  20. xv) SEQ ID NOS: 102, 103 and 104;

Description

LINK TO RELATED APPLICATION [0001] This application claims the benefit of U.S. Provisional Patent Application No. 63/043,114, filed June 23, 2020, and U.S. Provisional Patent Application No. 63/111,459, filed November 9, 2020, which are incorporated herein by reference in their entireties. AREAS OF TECHNOLOGY [0002] The present invention relates generally to the field of molecular biology and medicine. More specifically, the present invention provides fusion proteins comprising an antibody to PD-1 or an antigen-binding fragment thereof linked to an IL-15 polypeptide that is in turn linked to an IL-15 receptor alpha subunit (IL-15Rα) polypeptide comprising the Sushi domain of IL-15Rα. Furthermore, the present invention provides antibodies and antigen-binding fragments thereof that specifically bind to PD-1, as well as fusion proteins comprising such antibodies to PD-1 and their PD-1-binding fragments. Therapeutic compositions comprising the fusion antibodies or antibodies for the treatment of a disease are also disclosed. LEVEL OF TECHNOLOGY [0003] PD-1 is a 55 kDa, 286-amino acid, type I transmembrane protein that contains a membrane-proximal immunoreceptor tyrosine-based inhibitory motif (ITIM) and a membrane-distal tyrosine-based switch motif (ITSM). The cytosolic domain of PD-1 contains two tyrosines, with the closest tyrosine (VAYEEL in mouse PD-1) located within the ITIM. Human PD-1 and mouse PD-1 share approximately 60% amino acid identity, with four potential N-glycosylation sites and residues that define the Ig-V domain being conserved. ITIM-like motifs surrounding the ITIM and the C-terminal tyrosine in the cytoplasmic region are also conserved between the human and mouse orthologs. PD-1 is a member of the CD-28 receptor family that is predominantly expressed on mature T cells in peripheral tissues and the tumor microenvironment. PD-1 is also expressed on other cell subsets other than T cells, including B cells, professional antigen-presenting cells (APCs), and natural killer (NK) cells. [0004] The ligands for PD-1 are members of the B7 family, PD-L1 (also known as B7-H1 and CD274) and PD-L2 (also known as B7-DC and CD273). PD-L1 is predominantly expressed in both lymphoid and non-lymphoid tissues, such as on CD4 and CD8 T cells, macrophage-lineage cells, in peripheral tissues, as well as on tumor cells, virus-infected cells, and autoimmune tissue cells. However, PD-L2 is characterized by a more limited expression than PD-L1, being expressed only on macrophages and activated dendritic cells. PD-1 ligands are expressed in many forms of human cancer, including melanoma, glioma, non-small cell lung cancer, squamous cell carcinoma of the head and neck, leukemia, pancreatic cancer, renal cell carcinoma, and hepatocellular carcinoma, and can be induced in virtually all forms of cancer. Interaction between PD-1 and its ligands leads to dephosphorylation and inactivation of the T-cell kinase ZAP70 and recruitment of SHP2. SHP2 directly dephosphorylates PI3K, which inhibits subsequent activation of Akt, causing a decrease in the number of tumor-infiltrating lymphocytes and a reduction in T-cell receptor-mediated proliferation, leading to immune evasion. Inhibition of the interaction between PD-1 and PD-L1 reverses immunosuppression, and the effect may be additive when blocking the interaction of PD-1 with PD-L2. [0005] Several commercial anti-PD-1 antibodies are currently used to treat diseases that involve PD-1-mediated immunosuppression. However, only a fraction of patients respond well to these therapies. Depending on the indication and other factors, the typical response rate in patients to each monotherapy ranges from 10% to 30%. One possible reason for this lack of response may be the absence of an inflammatory tumor microenvironment (TME), including activated anti-tumor CD8 + and CD4 + T cells, as well as other effectors such as NK cells. [0006] Combination therapies that utilize anti-PD-1 antibodies with various cytokines, such as IL-2, IL-15, IL-21, tumor necrosis factor (TNF), and granulocyte-macrophage colony-stimulating factor (GM-CSF), may have some efficacy in the treatment of cancer and infection. However, such therapies are limited by systemic toxicity associated with both the high circulating cytokine concentrations required for efficacy and the lack of specificity of the administered cytokine for the affected cells and tissues. [0007] IL-15 is a 12.5 kDa glycoprotein containing 114 amino acids and belongs to the four-helix bundle cytokine family, which also includes IL-2, IL-4, IL-7, IL-9, granulocyte colony-stimulating factor (G-CSF), and GM-CSF. IL-15 Secreted by macrophages, dendritic cells, and monocytes, IL-15 can stimulate central memory CD8 T cells to exert immunity without modulating effects on other T cells. IL-15 can also activate NK cells, effector CD8 T cells, and memory CD8 T cells, as well as rescue T cells from regulatory T cell (Treg)-induced apoptosis. IL-15 administration