WO2022148279A1 - Produit pharmaceutique et son utilisation - Google Patents

Produit pharmaceutique et son utilisation Download PDF

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Publication number
WO2022148279A1
WO2022148279A1 PCT/CN2021/142299 CN2021142299W WO2022148279A1 WO 2022148279 A1 WO2022148279 A1 WO 2022148279A1 CN 2021142299 W CN2021142299 W CN 2021142299W WO 2022148279 A1 WO2022148279 A1 WO 2022148279A1
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seq
amino acid
acid sequence
antigen
set forth
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PCT/CN2021/142299
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English (en)
Chinese (zh)
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张欣
丁健
马慧
潘晓龙
傅士龙
周立耀
袁洋洋
马树立
赵猛
师二侠
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苏州丁孚靶点生物技术有限公司
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Priority to CN202180089662.9A priority Critical patent/CN116761622A/zh
Publication of WO2022148279A1 publication Critical patent/WO2022148279A1/fr

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    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants

Definitions

  • the present application relates to the field of biomedicine, in particular to a pharmaceutical product and use thereof.
  • GITR is a member of the TNF receptor family.
  • GITR is a type I transmembrane protein with a molecular weight of approximately 26 kDa and shares 14-28% homology with other molecules of the TNF receptor family (Gurney AL et al, 1999).
  • GITR is constitutively high on Treg and low on unactivated CD4 + T cells, CD8 + T cells, NK and NKT cells.
  • When T cells are activated GITR is highly expressed in CD4 + T, CD8 + T and NK cells (S. Ronchetti, et al., 2004).
  • the natural ligand of GITR, GITRL is expressed on APC. After GITRL binds to GITR, it provides a costimulatory signal, modulates antigen-specific T cell responses, and enhances cellular and humoral immunity (Selvakumar Sukumar et al., 2017).
  • the application provides a pharmaceutical product comprising an antigen binding protein targeting GITR and an immune checkpoint inhibitor.
  • the pharmaceutical product has one or more beneficial effects in the following group: 1) effectively preventing, relieving or treating cancer; 2) synergistic effect of the GITR-targeting antigen binding protein and the immune checkpoint inhibitor.
  • the present application also provides the use and pharmaceutical use of the pharmaceutical product in preventing, alleviating or treating cancer, especially GITR-positive cancer and/or PD-1-positive cancer.
  • the present application also provides the use of the antigen-binding protein targeting GITR in preventing, relieving or treating cancer, especially GITR-positive cancer.
  • the application provides a pharmaceutical product comprising an antigen-binding protein targeting GITR and an immune checkpoint inhibitor, wherein the antigen-binding protein comprises at least one of the VHs whose amino acid sequence is shown in SEQ ID NO: 23 one CDR and comprising at least one CDR in VL whose amino acid sequence is set forth in SEQ ID NO:24.
  • the pharmaceutical product is a pharmaceutical composition.
  • the antigen binding protein comprises HCDR3 in VH having the amino acid sequence set forth in SEQ ID NO:23.
  • the antigen binding protein comprises HCDR2 in VH having the amino acid sequence set forth in SEQ ID NO:23.
  • the antigen binding protein comprises HCDR1 in VH having the amino acid sequence set forth in SEQ ID NO:23.
  • the antigen binding protein comprises LCDR1 in VL having the amino acid sequence set forth in SEQ ID NO:24.
  • the antigen binding protein comprises LCDR2 in VL having the amino acid sequence set forth in SEQ ID NO:24.
  • the antigen binding protein comprises LCDR3 in VL having the amino acid sequence set forth in SEQ ID NO:24.
  • the antigen binding protein comprises HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:2.
  • the antigen binding protein comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:4.
  • the antigen binding protein comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:3.
  • the antigen binding protein comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:12.
  • the antigen binding protein comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:11.
  • the antigen binding protein comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:10.
  • the antigen-binding protein comprises an antibody or antigen-binding fragment thereof.
  • the antigen-binding fragment comprises Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
  • the antibody is a humanized antibody.
  • the VL includes framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
  • the C-terminus of the L-FR1 is directly or indirectly linked to the N-terminus of the LCDR1, and the L-FR1 comprises the amino acid sequence set forth in SEQ ID NO:39.
  • the L-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 13, 30, 34.
  • the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 comprises the amino acid sequence set forth in SEQ ID NO:40.
  • the L-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 14, 31.
  • the L-FR3 is located between the LCDR2 and the LCDR3, and the L-FR3 comprises the amino acid sequence set forth in SEQ ID NO:41.
  • the L-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 15, 32.
  • the N-terminus of the L-FR4 is linked to the C-terminus of the LCDR3, and the L-FR4 comprises the amino acid sequence set forth in SEQ ID NO:42.
  • the L-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 16, 33.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO:24.
  • the VL comprises the amino acid sequence set forth in any one of SEQ ID Nos: 9, 20-22.
  • the antigen binding protein comprises an antibody light chain constant region
  • the antibody light chain constant region comprises a human IgK constant region
  • the antibody light chain constant region comprises the amino acid sequence set forth in SEQ ID NO:52.
  • the antigen binding protein comprises an antibody light chain LC
  • the LC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 50, 46-48.
  • the VH includes framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
  • the C-terminus of the H-FR1 is directly or indirectly linked to the N-terminus of the HCDR1, and the H-FR1 comprises the amino acid sequence set forth in SEQ ID NO:35.
  • the H-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 5, 25, 29.
  • the H-FR2 is located between the HCDR1 and the HCDR2, and the H-FR2 comprises the amino acid sequence set forth in SEQ ID NO:36.
  • the H-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 6, 26.
  • the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 comprises the amino acid sequence set forth in SEQ ID NO:37.
  • the H-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 7, 27.
  • the N-terminus of the H-FR4 is linked to the C-terminus of the HCDR3, and the H-FR4 comprises the amino acid sequence set forth in SEQ ID NO:38.
  • the H-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 8, 28.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO:23.
  • the VH comprises the amino acid sequence set forth in any one of SEQ ID Nos: 1, 17-19.
  • the antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgG heavy chain constant region.
  • the antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgGl heavy chain constant region.
  • the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:51.
  • the antigen binding protein comprises an antibody heavy chain HC
  • the HC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 49, 43-45.
  • the immune checkpoint inhibitor blocks the interaction of PD-1 and PD-L1.
  • the immune checkpoint inhibitor comprises a PD-1 inhibitor.
  • the immune checkpoint inhibitor comprises a PD-1 antibody or antigen-binding fragment thereof.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR3, wherein the HCDR3 of the PD-1 antibody or antigen-binding fragment comprises the HCDR3 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR2, wherein the HCDR2 of the PD-1 antibody or antigen-binding fragment comprises the HCDR2 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR1, wherein the HCDR1 of the PD-1 antibody or antigen-binding fragment comprises the HCDR1 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR3, wherein the LCDR3 of the PD-1 antibody or antigen-binding fragment comprises the LCDR3 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR2, wherein the LCDR2 of the PD-1 antibody or antigen-binding fragment comprises the LCDR2 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR1, wherein the LCDR1 of the PD-1 antibody or antigen-binding fragment comprises LCDR1 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises a VH comprising the VH of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises a VL comprising the VL of RMP1-14.
  • the GITR-targeting antigen binding protein and the immune checkpoint inhibitor are immiscible with each other in the drug product.
  • the GITR-targeting antigen binding protein and the immune checkpoint inhibitor are present in separate containers in the drug product.
  • the application provides the use of the pharmaceutical product described in the application in the preparation of a medicament for preventing, relieving or treating cancer.
  • the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
  • the cancer comprises a solid tumor.
  • the cancer is selected from the group consisting of colon cancer, rectal cancer, and breast cancer.
  • the present application provides the use of an antigen binding protein targeting GITR and an immune checkpoint antagonist in the preparation of a medicament for preventing, relieving or treating cancer, wherein the antigen binding protein comprises an amino acid sequence as shown in SEQ ID NO:23 at least one CDR in the VH shown and comprising at least one CDR in the VL whose amino acid sequence is shown in SEQ ID NO:24.
  • the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
  • the cancer comprises a solid tumor.
  • the cancer is selected from the group consisting of colon cancer, rectal cancer, and breast cancer.
  • the antigen binding protein comprises HCDR3 in VH having the amino acid sequence set forth in SEQ ID NO:23.
  • the antigen binding protein comprises HCDR2 in VH having the amino acid sequence set forth in SEQ ID NO:23.
  • the antigen binding protein comprises HCDR1 in VH having the amino acid sequence set forth in SEQ ID NO:23.
  • the antigen binding protein comprises LCDR1 in VL having the amino acid sequence set forth in SEQ ID NO:24.
  • the antigen binding protein comprises LCDR2 in VL having the amino acid sequence set forth in SEQ ID NO:24.
  • the antigen binding protein comprises LCDR3 in VL having the amino acid sequence set forth in SEQ ID NO:24.
  • the antigen binding protein comprises HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:2.
  • the antigen binding protein comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:4.
  • the antigen binding protein comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:3.
  • the antigen binding protein comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:12.
  • the antigen binding protein comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:11.
  • the antigen binding protein comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:10.
  • the antigen-binding protein comprises an antibody or antigen-binding fragment thereof.
  • the antigen-binding fragment comprises Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
  • the antibody is a humanized antibody.
  • the VL includes framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
  • the C-terminus of the L-FR1 is directly or indirectly linked to the N-terminus of the LCDR1, and the L-FR1 comprises the amino acid sequence set forth in SEQ ID NO:39.
  • the L-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 13, 30, 34.
  • the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 comprises the amino acid sequence set forth in SEQ ID NO:40.
  • the L-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 14, 31.
  • the L-FR3 is located between the LCDR2 and the LCDR3, and the L-FR3 comprises the amino acid sequence set forth in SEQ ID NO:41.
  • the L-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 15, 32.
  • the N-terminus of the L-FR4 is linked to the C-terminus of the LCDR3, and the L-FR4 comprises the amino acid sequence set forth in SEQ ID NO:42.
  • the L-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 16, 33.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO:24.
  • the VL comprises the amino acid sequence set forth in any one of SEQ ID Nos: 9, 20-22.
  • the antigen binding protein comprises an antibody light chain constant region
  • the antibody light chain constant region comprises a human IgK constant region
  • the antibody light chain constant region comprises the amino acid sequence set forth in SEQ ID NO:52.
  • the antigen binding protein comprises an antibody light chain LC
  • the LC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 50, 46-48.
  • the VH includes framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
  • the C-terminus of the H-FR1 is directly or indirectly linked to the N-terminus of the HCDR1, and the H-FR1 comprises the amino acid sequence set forth in SEQ ID NO:35.
  • the H-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 5, 25, 29.
  • the H-FR2 is located between the HCDR1 and the HCDR2, and the H-FR2 comprises the amino acid sequence set forth in SEQ ID NO:36.
  • the H-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 6, 26.
  • the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 comprises the amino acid sequence set forth in SEQ ID NO:37.
  • the H-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 7, 27.
  • the N-terminus of the H-FR4 is linked to the C-terminus of the HCDR3, and the H-FR4 comprises the amino acid sequence set forth in SEQ ID NO:38.
  • the H-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 8, 28.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO:23.
  • the VH comprises the amino acid sequence set forth in any one of SEQ ID Nos: 1, 17-19.
  • the antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgG heavy chain constant region.
  • the antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgGl heavy chain constant region.
  • the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:51.
  • the antigen binding protein comprises an antibody heavy chain HC
  • the HC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 49, 43-45.
  • the immune checkpoint inhibitor blocks the interaction of PD-1 and PD-L1.
  • the immune checkpoint inhibitor comprises a PD-1 inhibitor.
  • the immune checkpoint inhibitor comprises a PD-1 antibody or antigen-binding fragment thereof.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR3, wherein the HCDR3 of the PD-1 antibody or antigen-binding fragment comprises the HCDR3 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR2, wherein the HCDR2 of the PD-1 antibody or antigen-binding fragment comprises the HCDR2 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR1, wherein the HCDR1 of the PD-1 antibody or antigen-binding fragment comprises the HCDR1 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR3, wherein the LCDR3 of the PD-1 antibody or antigen-binding fragment comprises the LCDR3 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR2, wherein the LCDR2 of the PD-1 antibody or antigen-binding fragment comprises the LCDR2 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR1, wherein the LCDR1 of the PD-1 antibody or antigen-binding fragment comprises LCDR1 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises a VH comprising the VH of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises a VL comprising the VL of RMP1-14.
  • the present application provides a method for preventing, relieving or treating cancer, comprising the steps of: administering the antigen-binding protein described in the present application to a subject in need, and/or, to a subject in need are administered the immune checkpoint inhibitors described in this application.
  • the method comprises the step of administering to a subject in need thereof a pharmaceutical product described herein.
  • the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
  • the cancer comprises a solid tumor.
  • the cancer is selected from the group consisting of colon cancer, rectal cancer, and breast cancer.
  • the antigen binding protein is administered at a dose of 1 mg/Kg to 10 mg/Kg.
  • the immune checkpoint inhibitor is administered at a dose of 1 mg/Kg to 10 mg/Kg.
  • the frequency of administration of the antigen binding protein is twice a week.
  • the frequency of administration of the immune checkpoint inhibitor is twice a week.
  • the antigen binding protein is administered by injection.
  • the immune checkpoint inhibitor is administered by injection.
  • the antigen binding protein is at a concentration of 1 mg/mL to 10 mg/mL.
  • the concentration of the immune checkpoint inhibitor is 1 mg/mL to 10 mg/mL.
  • Figure 1A shows the effect of the drug product described in this application on the body weight of mice.
  • Figure 1B shows the effect of the drug product described herein on tumor volume in mice.
  • Figures 2A-2D show the effect of different treatment groups on tumor volume in mice.
  • Figure 3A shows the effect of the GITR-targeting antigen binding proteins described herein on the body weight of mice.
  • Figure 3B shows the effect of the antigen-binding protein targeting GITR of the present application on tumor volume in mice.
  • Figures 4A-4D show the effect of different treatment groups on tumor volume in mice.
  • Figures 5A-5B show the effect of the drug products described herein on mouse body weight and mouse tumor volume.
  • antigen-binding protein generally refers to a protein comprising an antigen-binding moiety, and optionally a scaffold or backbone moiety that allows the antigen-binding moiety to adopt a conformation that facilitates the binding of the antigen-binding protein to the antigen.
  • antigen binding proteins include, but are not limited to, antibodies, antigen binding fragments (Fab, Fab', F(ab) 2 , Fv fragments, F(ab') 2 , scFv, di-scFv and/or dAb), immunoconjugation antibodies, multispecific antibodies (eg, bispecific antibodies), antibody fragments, antibody derivatives, antibody analogs, or fusion proteins, etc., as long as they exhibit the desired antigen-binding activity.
  • Fab antigen binding fragments
  • Fv fragments F(ab') 2
  • scFv di-scFv and/or dAb
  • immunoconjugation antibodies eg, multispecific antibodies (eg, bispecific antibodies), antibody fragments, antibody derivatives, antibody analogs, or fusion proteins, etc., as long as they exhibit the desired antigen-binding activity.
  • the "isolated antigen-binding protein” may comprise an antigen-binding moiety and, optionally, a scaffold or framework moiety that allows the antigen-binding moiety to adopt a conformation that facilitates binding of the antigen-binding moiety to the antigen.
  • the isolated antigen binding protein may comprise, for example, an antibody-derived protein scaffold or an alternative protein scaffold or artificial scaffold with grafted CDRs or CDR derivatives.
  • Such scaffolds may include mutated antibody-derived scaffolds introduced, for example, to stabilize the three-dimensional structure of the antigen binding protein, as well as fully synthetic scaffolds comprising, for example, biocompatible polymers.
  • the isolated antigen binding protein can bind to GITR protein derived from human and monkey with a KD value of 7x10-12 or lower, wherein the KD value can be determined by BLI method.
  • GITR generally refers to "glucocorticoid-induced TNF-related gene", also known as TNF receptor superfamily 18 (TNFRSF18), AITR or CD357.
  • the GITR may bind to a GITR ligand (GITRL).
  • GITRL GITR ligand
  • the GITR can include any variant or isoform of the GITR that is naturally expressed by the cell.
  • GITR of human origin can include 3 variants whose amino acid sequences can be shown in Accession Nos. NP_004186, NP_683699 and NP_683700, respectively.
  • GITR The amino acid and nucleic acid sequences of human and murine forms of GITR are described in WO98/06842, which is incorporated herein by reference. See also GenBank Accession Nos. Q9Y5U5 (amino acid sequence of human origin) and AF109216 (nucleic acid and amino acid sequence of murine origin).
  • a mature human GITR polypeptide may comprise the amino acid sequence set forth in SEQ ID NO:49.
  • An exemplary mature GITR protein from cynomolgus monkey can comprise the amino acid sequence set forth in SEQ ID NO:51.
  • the term "GITR" also includes naturally occurring alleles.
  • GITRL generally refers to a GITR ligand or functionally active portion thereof, and/or a soluble GITR ligand.
  • GITRL can also include naturally-occurring allelic variants of GITRL, variants of GITR ligands that differ in amino acid sequence from naturally-occurring GITR ligand molecules, and combinations of such variants, wherein these variants retain specificity the ability to bind to the GITR receptor.
  • the term "immune checkpoint inhibitor” generally refers to any agent capable of inhibiting the biological activity of an immune checkpoint.
  • the immune checkpoint may be a regulatory molecule that plays an inhibitory role in the immune system.
  • the immune checkpoint can be expressed on immune cells to inhibit the function of immune cells.
  • the immune checkpoints can be associated with tumor immune escape.
  • the immune checkpoint may include PD-1.
  • the inhibitor can include any agent capable of inhibiting the interaction between a ligand and a receptor (e.g., PD-1 and PD-L1).
  • the reagents can be chemical reagents or proteins and/or polypeptides.
  • the agent may comprise an antibody or antigen-binding fragment thereof.
  • the term "PD-1" generally refers to the CD28 family of immunosuppressive receptors. PD-1 can be expressed on preactivated T cells and bind to two ligands, PD-L1 and/or PD-L2.
  • the PD-1 may include human PD-1 (hPD-1) or its variants, isoforms, and species homologues, and analogs that share at least one epitope with hPD-1.
  • the GenBank accession number for hPD-1 is U64863.
  • the term "PD-L1" generally refers to one of the cell surface glycoprotein ligands of PD-1.
  • the PD-L1 can bind to PD-1.
  • the PD-L1 can downregulate T cell activation and/or cytokine secretion when combined with PD-1.
  • the PD-L1 may include human PD-L (hPD-L1) or variants, isoforms and species homologues thereof, and analogs having at least one common epitope with hPD-L1.
  • the GenBank accession number of hPD-L1 is Q9NZQ7.
  • the term "pharmaceutical product” generally refers to a substance used to prevent, alleviate or treat human diseases (eg cancer), purposefully modulate human physiology, and specify indications or functional indications, usage and dosage.
  • the drug product may include a biological product.
  • the drug product can include an antigen binding protein.
  • the pharmaceutical product may include a chemical.
  • the drug product may be a mixture or a composition.
  • the pharmaceutical product may exist in the form of an integral package, or may be packaged into multiple packages.
  • GITR-positive cancer generally refers to cancers in which tumor cells (eg, tumor-infiltrating Treg cells) express GITR.
  • PD-1 positive cancer generally refers to cancers in which tumor cells express PD-1.
  • solid tumor generally refers to a tangible tumor that can be detected by means of clinical examination (eg, X-ray irradiation, CT scan, B-ultrasound or palpation, etc.).
  • the tumor may comprise a neoplasm or solid lesion formed by abnormal cell growth.
  • breast cancer generally refers to cancer that occurs in the epithelial tissue of the breast glands.
  • rectal cancer generally refers to cancer that occurs from the dentate line to the rectosigmoid junction.
  • colon cancer generally refers to cancer of the digestive tract that occurs in the colon.
  • the colon cancer can occur at the junction of the rectum and sigmoid colon.
  • variable domain generally refers to the amino-terminal domain of an antibody heavy or light chain.
  • the variable domains of heavy and light chains may be referred to as “VH” and “VL”, respectively. These domains are usually the most variable part of the antibody (relative to other antibodies of the same type) and contain the antigen binding site.
  • variable generally refers to the fact that certain segments of the variable domains differ greatly in sequence between antibodies.
  • the V domain mediates antigen binding and determines the specificity of a particular antibody for its particular antigen.
  • CDRs or HVRs hypervariable regions
  • the more highly conserved portions of variable domains are referred to as framework regions (FRs).
  • the variable domains of native heavy and light chains may each comprise four FR regions, most adopting a beta-sheet configuration, connected by three CDRs that form loops connecting, and in some cases forming, the ⁇ -sheet structure. part.
  • the CDRs in each chain can be held in close proximity by the FR regions, and the CDRs from the other chain together contribute to the formation of the antigen-binding site of the antibody (see Kabat et al, Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991)).
  • the constant domains may not be directly involved in the binding of the antibody to the antigen, but exhibit various effector functions, such as the involvement of the antibody in antibody-dependent cellular cytotoxicity.
  • antibody generally refers to an immunoglobulin or fragment or derivative thereof, and encompasses any polypeptide that includes an antigen-binding site, whether produced in vitro or in vivo.
  • the term may include polyclonal, monoclonal, monospecific, multispecific, nonspecific, humanized, single-stranded, chimeric, synthetic, recombinant, hybrid, mutant and transplanted antibodies.
  • the term “antibody” also includes antibody fragments, such as Fab, F(ab')2, Fv, scFv, Fd, dAbs and other antibody fragments that retain antigen binding function (eg, specifically bind GITR). Typically, such fragments may include an antigen binding domain.
  • the basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains.
  • IgM antibody is composed of 5 basic heterotetrameric units and another polypeptide called J chain, and contains 10 antigen-binding sites, while IgA antibody includes 2-5 that can be combined with J chain to form multivalent Combined basic 4-chain unit.
  • the 4-chain unit is typically about 150,000 Daltons.
  • Each L chain is connected to the H chain by one covalent disulfide bond, while the two H chains are connected to each other by one or more disulfide bonds depending on the isotype of the H chain.
  • Each H and L chain also has regularly spaced intrachain disulfide bridges.
  • Each H chain has a variable domain (VH) at the N-terminus, followed by three constant domains (CH) for each of the alpha and gamma chains and four CH domains for the mu and epsilon isoforms.
  • Each L chain has a variable domain (VL) at the N-terminus and a constant domain at the other end.
  • VL corresponds to VH and CL corresponds to the first constant domain (CH1) of the heavy chain.
  • Particular amino acid residues can be considered to form the interface between the light and heavy chain variable domains.
  • VH and VL can pair together to form a single antigen binding site.
  • immunoglobulins from any vertebrate species can be classified into one of two distinct types, called kappa and lambda, based on the amino acid sequence of their constant domains. Depending on the amino acid sequence of their heavy chain (CH) constant domains, immunoglobulins can be divided into different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated alpha, delta, epsilon, gamma, and mu, respectively.
  • the gamma and alpha classes are further divided into subclasses based on relatively small differences in CH sequence and function, eg, humans can express the following subclasses: IgGl, IgG2A, IgG2B, IgG3, IgG4, IgAl and IgKl.
  • CDR generally refers to regions of antibody variable domains whose sequences are highly variable and/or form structurally defined loops.
  • an antibody may include six CDRs; three in the VH (HCDRl, HCDR2, HCDR3), and three in the VL (LCDRl, LCDR2, LCDR3).
  • HCDR3 and LCDR3 can display most of the diversity of the six CDRs, and HCDR3 in particular is thought to play a unique role in conferring fine specificity to antibodies.
  • variable domains of native heavy and light chains may each comprise four FR regions, namely four in VH (H-FR1, H-FR2, H-FR3, and H-FR4), and in VL Four (L-FR1, L-FR2, L-FR3, and L-FR4).
  • VL of the isolated antigen binding proteins described herein can include the framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
  • the VH of the isolated antigen binding proteins described herein can include the framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
  • the term "antigen-binding fragment” generally refers to a fragment having antigen-binding activity.
  • the antigen-binding fragment may include Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
  • Fab generally refers to a fragment containing the variable domain of the heavy chain and the variable domain of the light chain, and also containing the constant domain of the light chain and the first constant domain (CH1) of the heavy chain
  • Fab' generally refers to a fragment that differs from Fab by adding a small number of residues (including one or more cysteines from the antibody hinge region) to the carboxy terminus of the heavy chain CH1 domain
  • F(ab"') 2 generally refers to a dimer of Fab', an antibody fragment comprising two Fab fragments linked by a disulfide bridge on the hinge region.
  • Fv generally refers to the smallest antibody fragment containing the entire antigen recognition and binding site.
  • the fragment may consist of a heavy chain variable region and a light chain variable region in a tightly non-covalently bound dimer;
  • dsFv generally refers to disulfide-stabilized Fv fragments, The bond between its single light chain variable region and single heavy chain variable region is a disulfide bond.
  • dAb fragment generally refers to antibody fragments consisting of VH domains.
  • scFv generally refers to a monovalent molecule formed by covalently linking and pairing one heavy chain variable domain and one light chain variable domain of an antibody through a flexible peptide linker; such scFv molecules may have a general Structure: NH2 -VL-Linker-VH-COOH or NH2 -VH-Linker-VL-COOH.
  • humanized antibody generally refers to an antibody in which some or all of the amino acids other than the CDR regions of a non-human antibody (eg, a mouse antibody) have been replaced by corresponding amino acids derived from human immunoglobulins. Small additions, deletions, insertions, substitutions or modifications of amino acids in the CDR regions are also permissible, so long as they still retain the ability of the antibody to bind to a particular antigen.
  • a humanized antibody may optionally comprise at least a portion of a human immunoglobulin constant region.
  • a "humanized antibody” retains antigenic specificity similar to the original antibody.
  • “Humanized” forms of non-human (eg, murine) antibodies may minimally comprise chimeric antibodies that contain sequences derived from non-human immunoglobulins.
  • CDR region residues in a human immunoglobulin can be substituted with a non-human species (donor antibody) (such as mouse, rat) having the desired properties, affinity and/or ability , rabbit or non-human primate) CDR region residue replacement.
  • donor antibody such as mouse, rat
  • FR region residues of the human immunoglobulin can be replaced with corresponding non-human residues.
  • humanized antibodies may contain amino acid modifications that are not present in the recipient antibody or in the donor antibody. These modifications may be made to further improve antibody properties, such as binding affinity.
  • the term “directly connected” may be opposed to the term “indirectly connected”, which generally refers to a direct connection.
  • the direct connection may be the case where substances are directly connected without a spacer.
  • the spacer may be a linker.
  • the linker can be a peptide linker.
  • the term “indirectly connected” generally refers to a situation where substances are not directly connected.
  • the indirect linkage may be the case of linkage through a spacer.
  • the C-terminus of L-FR1 and the N-terminus of LCDR1 may be linked directly or indirectly.
  • isolated nucleic acid molecule generally refers to any length of nucleotides in isolated form, deoxyribonucleotides or ribonucleotides, or analogs isolated from their natural environment or synthetically synthesized.
  • the term "vector” generally refers to a nucleic acid delivery vehicle into which a polynucleotide encoding a protein can be inserted and the protein can be expressed.
  • a vector can be expressed by transforming, transducing or transfecting a host cell so that the genetic material elements it carries are expressed in the host cell.
  • the vector can be a plasmid.
  • a vector may contain various elements that control expression, including promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. Additionally, the vector may also contain an origin of replication site.
  • the carrier may also include components to assist its entry into the cell, such as viral particles, liposomes or protein coats, but not only these substances.
  • the term "cell” generally refers to a single cell, cell line, or cell culture that can be or has been the recipient of a subject plasmid or vector, comprising a nucleic acid molecule described herein or a nucleic acid molecule described herein Carrier.
  • a cell can include the progeny of a single cell. Progeny may not necessarily be identical (in morphology or in genome) to the original parent cell due to natural, accidental or intentional mutation.
  • Cells can include cells transfected in vitro with the vectors described herein.
  • the cells can be bacterial cells (eg, E. coli), yeast cells, or other eukaryotic cells, eg, the cells can be mammalian cells.
  • the term "pharmaceutical composition” generally refers to a composition suitable for administration to a patient, eg, a human patient.
  • a pharmaceutical composition described herein which may comprise an isolated antigen binding protein described herein, a nucleic acid molecule described herein, a vector described herein, and/or a cell described herein, and Optionally a pharmaceutically acceptable adjuvant.
  • the pharmaceutical composition may also comprise one or more (pharmaceutically effective) carriers, stabilizers, excipients, diluents, solubilizers, surfactants, emulsifiers and/or suitable preservatives preparation. Acceptable ingredients of the compositions may be nontoxic to recipients at the dosages and concentrations employed.
  • compositions of the present application include, but are not limited to, liquid, frozen, and lyophilized compositions.
  • pharmaceutically acceptable adjuvant generally refers to any and all solvents, dispersion media, coatings, isotonic and absorption delaying agents, etc. that are compatible with pharmaceutical administration and are generally safe and non-toxic , and is neither biologically nor otherwise undesirable.
  • the term "subject” generally refers to a human or non-human animal, including but not limited to cats, dogs, horses, pigs, cows, sheep, rabbits, mice, rats or monkeys.
  • the term "about” generally refers to a range of 0.5%-10% above or below the specified value, such as 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%.
  • Antigen-binding proteins targeting GITR Antigen-binding proteins targeting GITR
  • the antigen binding protein targeting GITR involved in the present application may comprise at least one CDR in the VH whose amino acid sequence is shown in SEQ ID NO: 23.
  • the antigen binding protein targeting GITR described in the present application can comprise HCDR3 in VH whose amino acid sequence is shown in SEQ ID NO: 23.
  • the antigen binding protein targeting GITR described in the present application may comprise HCDR2 in VH whose amino acid sequence is shown in SEQ ID NO: 23.
  • the antigen-binding protein targeting GITR described in the present application may comprise HCDR1 in the VH whose amino acid sequence is shown in SEQ ID NO: 23.
  • the CDRs in the VL and/or VH of the antigen binding protein targeting GITR described in the present application can be defined according to Kabat.
  • the antigen binding protein targeting GITR described in the present application may comprise at least one CDR in VL whose amino acid sequence is shown in SEQ ID NO: 24.
  • the antigen binding protein targeting GITR described in the present application may comprise LCDR1 in VL whose amino acid sequence is as shown in SEQ ID NO:24.
  • the antigen binding protein targeting GITR described in the present application may comprise LCDR2 in VL whose amino acid sequence is as shown in SEQ ID NO:24.
  • the antigen-binding protein targeting GITR described in the present application may comprise LCDR3 in VL whose amino acid sequence is shown in SEQ ID NO: 24.
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 2.
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 3.
  • the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 4.
  • the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 10.
  • the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 11.
  • the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 12.
  • the antigen-binding proteins targeting GITR described in the present application may include antibodies or antigen-binding fragments thereof.
  • the antigen binding proteins targeting GITR described in this application may include, but are not limited to, recombinant antibodies, monoclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, bispecific antibodies, single chain antibodies, diabodies, Tribodies, tetrabodies, Fv fragments, scFv fragments, Fab fragments, Fab' fragments, F(ab')2 fragments and camelized single domain antibodies.
  • the antibody may be a humanized antibody.
  • the GITR-targeting antigen-binding proteins described herein can be immunospecifically bound to a relevant antigen (eg, human GITR) and comprise a framework (FR) region having substantially the amino acid sequence of a human antibody and An antibody or a variant, derivative, analog or fragment thereof having substantially the complementarity determining regions (CDRs) of the amino acid sequence of a non-human antibody.
  • CDRs complementarity determining regions
  • the humanized antibody may comprise substantially all at least one and usually both variable domains (Fab, Fab', F(ab')2, FabC, Fv), wherein all or substantially all CDR regions correspond to non-human
  • the CDR regions and all or substantially all framework regions of an immunoglobulin are framework regions having human immunoglobulin consensus sequences.
  • a humanized antibody also comprises at least a portion of an immunoglobulin constant region (eg, Fc), typically a human immunoglobulin constant region.
  • a humanized antibody contains at least the variable domains of a light chain as well as a heavy chain. Antibodies may also include the CH1, hinge, CH2, CH3, and CH4 regions of the heavy chain.
  • the humanized antibody contains only humanized light chains.
  • the humanized antibody contains only humanized heavy chains.
  • the humanized antibody contains only humanized variable domains of the light chain and/or the humanized heavy chain.
  • the antigen-binding fragment may include Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
  • the antigen-binding protein targeting GITR described in the present application can compete with a reference antibody for binding to the GITR protein, wherein the reference antibody can comprise HCDR1 in the VH whose amino acid sequence is shown in SEQ ID NO: 23 -3 and LCDR1-3 in VL whose amino acid sequence is shown in SEQ ID NO:24.
  • the HCDR1-3 of the reference antibody may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively, and the LCDR1-3 of the reference antibody
  • the amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 can be included, respectively.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 23; and the heavy chain variable region of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 24 amino acid sequence.
  • the light chain of the reference antibody may comprise the amino acid sequence shown in any one of SEQ ID NO:50, 46-48; and the heavy chain of the reference antibody may comprise SEQ ID NO:49 , the amino acid sequence shown in any one of 43-45.
  • the VL of the GITR-targeting antigen binding protein described herein may include framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
  • the C-terminus of the L-FR1 of the GITR-targeting antigen binding protein described in the present application may be directly or indirectly linked to the N-terminus of the LCDR1, and the L-FR1 may comprise the SEQ ID NO: 39. amino acid sequence shown.
  • the L-FR1 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 13, 30, and 34.
  • the L-FR2 of the GITR-targeting antigen binding protein described in the present application may be located between the LCDR1 and the LCDR2, and the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 40 .
  • WYLQX 1 PGQSPKLLIY (SEQ ID NO: 40), wherein X 1 can be R or K.
  • the L-FR2 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 14 and 31.
  • the L-FR3 of the GITR-targeting antigen binding protein described in the present application may be located between the LCDR2 and the LCDR3, and the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 41 .
  • GVPDRFSGSGSGTDFTLKISRVEAEDX 1 GVYYC (SEQ ID NO: 41), wherein X 1 can be L or V.
  • the L-FR3 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 15 and 32.
  • the N-terminus of the L-FR4 of the GITR-targeting antigen binding protein described in the present application can be linked to the C-terminus of the LCDR3, and the L-FR4 can comprise the amino acid shown in SEQ ID NO: 42 sequence.
  • FGGGTKX 1 EIK (SEQ ID NO: 42), wherein X 1 can be V or L.
  • the L-FR4 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 16 and 33.
  • the VL of the antigen binding protein targeting GITR described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 24.
  • DIVMTQSPLSLPVX 1 LGX 2 X 3 ASISCRSSQTIVHSNGNTYLEWYLQX 4 PGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDX 5 GVYYCFQGSHVPWTFGGGTKX 6 EIK (SEQ ID NO: 24), wherein X 1 can be S or T, X 2 can be Q or D, X 3 can be P or Q, X 4 can is R or K, X5 can be L or V, X6 can be V or L.
  • VL of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 9, 20-22.
  • the GITR-targeting antigen binding protein described herein may include an antibody light chain constant region, and the antibody light chain constant region may include a human Ig ⁇ constant region.
  • the antibody light chain constant region of the antigen-binding protein targeting GITR described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 52.
  • the antigen binding protein targeting GITR described in the present application may comprise an antibody light chain LC, and the LC may comprise the amino acid sequence shown in any one of SEQ ID NOs: 50, 46-48.
  • the VH of the GITR-targeting antigen binding protein described herein may include framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
  • the C-terminus of the H-FR1 of the GITR-targeting antigen-binding protein described in the present application is directly or indirectly connected to the N-terminus of the HCDR1, and the H-FR1 may comprise SEQ ID NO: 35. amino acid sequence.
  • X 1 can be T or Q
  • X 2 can be G or T
  • X 3 can be I or L
  • X4 can be L or V
  • X5 can be Q or K
  • X6 can be S or T
  • X7 can be S or T
  • X8 can be S or T.
  • the H-FR1 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 5, 25, and 29.
  • the H-FR2 of the GITR-targeting antigen binding protein described herein may be located between the HCDR1 and the HCDR2, and the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 36 .
  • WIRQPX 1 GKGLEWLVLI SEQ ID NO: 36
  • X 1 can be P or S.
  • the H-FR2 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 6 and 26.
  • the H-FR3 of the GITR-targeting antigen binding protein described herein may be located between the HCDR2 and the HCDR3, and the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO:37 .
  • LLTVX 1 KDTSX 2 NQVX 3 LX 4 IX 5 X 6 X 7 DX 8 X 9 DTATYYCAR (SEQ ID NO: 37), wherein X 1 can be S or T, X 2 can be N or K, and X 3 can be F or V, X 4 can be K or T, X 5 can be A or T, X 6 can be S or N, X 7 can be V or M, X 8 can be T or P, X 9 can be A or V .
  • the H-FR3 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 7 and 27.
  • the N-terminus of the H-FR4 of the GITR-targeting antigen binding protein described in the present application may be linked to the C-terminus of the HCDR3, and the H-FR4 may comprise the amino acid shown in SEQ ID NO: 38 sequence.
  • WGX 1 GTX 2 VTVSS (SEQ ID NO: 38), wherein X 1 can be Q or T and X 2 can be M or T.
  • the H-FR4 of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 8 and 28.
  • the VH of the antigen-binding protein targeting GITR described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 23.
  • X 1 can be T or Q
  • X 2 can be G or T
  • X 3 can be I or L
  • X 4 can be L or V
  • X 5 can be Q or K
  • X 6 can be S or T
  • X 7 can be S or T
  • X 8 can be S or T
  • X 9 can be P or S
  • X 10 can be S or T
  • X 11 can be N or K
  • X 12 can be F or V
  • X 13 can be K or T
  • X 14 can be A or T
  • X 15 can be S or N
  • X 16 can be V or M
  • the VH of the GITR-targeting antigen binding protein described herein may comprise the amino acid sequence shown in any one of SEQ ID NOs: 1, 17-19.
  • the GITR-targeting antigen-binding protein described herein may include an antibody heavy chain constant region, and the antibody heavy chain constant region may be derived from a human IgG heavy chain constant region.
  • the GITR-targeting antigen binding proteins described herein can include an antibody heavy chain constant region, and the antibody heavy chain constant region can be derived from a human IgGl heavy chain constant region.
  • the antibody heavy chain constant region of the antigen-binding protein targeting GITR described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 51.
  • the antigen-binding protein targeting GITR described in the present application may comprise an antibody heavy chain HC, and the HC may comprise the amino acid sequence shown in any one of SEQ ID NOs: 49, 43-45.
  • L-FR1 of the antigen binding protein targeting GITR described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 13
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 15
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 5.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:7.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:8.
  • L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO: 30, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 31, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:33, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:25. H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:26. L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27. L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO: 30
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14
  • L-FR3 Can comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 can comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 can comprise the amino acid sequence shown in SEQ ID NO:25.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:26.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO: 34
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:25.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:26.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO: 30, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 31, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:33, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29. H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6. L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27. L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO: 30, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29. H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6. L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27. L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO: 34
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the L-FR1 of the antigen binding protein targeting GITR described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 30, the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 31, the L-FR3 Can comprise the amino acid sequence shown in SEQ ID NO:32, L-FR4 can comprise the amino acid sequence shown in SEQ ID NO:33, and H-FR1 can comprise the amino acid sequence shown in SEQ ID NO:29.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO: 30, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29. H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6. L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27. L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • L-FR1 of the antigen binding protein targeting GITR described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 34
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the GITR-targeting antigen binding proteins described herein may comprise an antibody light chain variable region VL and an antibody heavy chain variable region VH.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:9
  • the VH can comprise the amino acid sequence set forth in SEQ ID NO:1.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:20 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:17.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:21 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:17.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:22 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:17.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:20
  • the VH can comprise the amino acid sequence set forth in SEQ ID NO:18.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:21 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:18.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:22 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:18.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:20 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:19.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:21 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:19.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:22 and the VH can comprise the amino acid sequence set forth in SEQ ID NO:19.
  • the GITR-targeting antigen binding proteins described herein may comprise an antibody light chain and an antibody heavy chain.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:50 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:49.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:46 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:43.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:47 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:43.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:48 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:43.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:46 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:44.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:47 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:44.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:48 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:44.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:46 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:45.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:47 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:45.
  • the light chain can comprise the amino acid sequence set forth in SEQ ID NO:48 and the heavy chain can comprise the amino acid sequence set forth in SEQ ID NO:45.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen-binding protein targeting GITR may comprise the amino acid sequence shown in SEQ ID NO: 13
  • the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 14
  • the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 14
  • the amino acid sequence shown in SEQ ID NO:15, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:5.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:7.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:8.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:9, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:1.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:50, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:49.
  • the antigen binding protein targeting GITR can be C3E2.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen-binding protein targeting GITR described in this application can comprise the amino acid sequence shown in SEQ ID NO:30
  • L-FR2 can comprise the amino acid sequence shown in SEQ ID NO:31
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:33
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:25.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:26.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:20, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:17.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:46, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:43.
  • the antigen binding protein targeting GITR can be 3E2 1-2.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO:30
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO:14
  • L-FR3 Can comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 can comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 can comprise the amino acid sequence shown in SEQ ID NO:25.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:26.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:21, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:17.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:47, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:43.
  • the antigen binding protein targeting GITR can be 3E2 1-3.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen-binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO:34
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO:14
  • L-FR3 Can comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 can comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 can comprise the amino acid sequence shown in SEQ ID NO:25.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:26.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:22, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:17.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:48, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:43.
  • the antigen binding protein targeting GITR can be 3E2 1-4.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen-binding protein targeting GITR described in this application can comprise the amino acid sequence shown in SEQ ID NO:30
  • L-FR2 can comprise the amino acid sequence shown in SEQ ID NO:31
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:33
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:20, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:18.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:46, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:44.
  • the antigen binding protein targeting GITR can be 3E2 2-2.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO:30
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO:14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:21, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:18.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:47, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:44.
  • the antigen binding protein targeting GITR can be 3E2 2-3.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen-binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO:34
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO:14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:22, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:18.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:48, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:44.
  • the antigen binding protein targeting GITR can be 3E2 2-4.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen-binding protein targeting GITR described in this application can comprise the amino acid sequence shown in SEQ ID NO:30
  • L-FR2 can comprise the amino acid sequence shown in SEQ ID NO:31
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:33
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:20, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:19.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:46, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:45.
  • the antigen binding protein targeting GITR can be 3E2 3-2.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO:30
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO:14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL may comprise the amino acid sequence set forth in SEQ ID NO:21, and the VH may comprise the amino acid sequence set forth in SEQ ID NO:19.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:47, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:45.
  • the antigen binding protein targeting GITR can be 3E2 3-3.
  • the HCDR1-3 of the antigen-binding protein targeting GITR may comprise the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and LCDR1-3 may The amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 are respectively included.
  • the L-FR1 of the antigen-binding protein targeting GITR described in this application may comprise the amino acid sequence shown in SEQ ID NO:34
  • L-FR2 may comprise the amino acid sequence shown in SEQ ID NO:14
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:32
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:16
  • H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:29.
  • H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:6.
  • L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:27.
  • L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:28.
  • the VL can comprise the amino acid sequence set forth in SEQ ID NO:22, and the VH can comprise the amino acid sequence set forth in SEQ ID NO:19.
  • the light chain may comprise the amino acid sequence set forth in SEQ ID NO:48, and the heavy chain may comprise the amino acid sequence set forth in SEQ ID NO:45.
  • the antigen binding protein targeting GITR can be 3E2 3-4.
  • the antigen binding protein targeting GITR may have one or more of the following properties: 1) capable of binding to GITR derived from human and monkey with a KD value of 7 ⁇ 10 ⁇ 12 or lower 2) can stimulate immune cell proliferation; 3) can stimulate immune cells to secrete IFN- ⁇ , and the secretion is measured in T cell activity assay; 4) can inhibit tumor growth and/or tumor cell proliferation; 5) can activate GITR signaling pathway; 6) can inhibit the combination of GITR and GITRL.
  • the antigen-binding protein targeting GITR can bind to GITR protein derived from human and monkey with a K D value of 7 ⁇ 10 ⁇ 12 M or lower, wherein the K D value can be determined by BLI method.
  • the KD value of the GITR-targeting antigen binding protein described in the present application for binding to a human-derived GITR protein may be ⁇ 7x10-12M , ⁇ 6x10-12M , ⁇ 5x10-12M , ⁇ 4x10-12M , ⁇ 3x10 -12 M, ⁇ 2x10 -12 M, ⁇ 1x10 -12 M, ⁇ 0.5x10 -12 M, ⁇ 0.1x10 -12 M, ⁇ 0.01x10 -12 M, ⁇ 0.05x10 -12 M, or ⁇ 0.001 x10-12M .
  • the K D value of the antigen binding protein targeting GITR described in the present application in combination with a monkey-derived GITR protein can be ⁇ 7 ⁇ 10 ⁇ 12 M, ⁇ 6 ⁇ 10 ⁇ 12 M, ⁇ 5 ⁇ 10 ⁇ 12 M, ⁇ 4 ⁇ 10 ⁇ 12 M, ⁇ 3x10 -12 M, ⁇ 2x10 -12 M, ⁇ 1x10 -12 M, ⁇ 0.5x10 -12 M, ⁇ 0.1x10 -12 M, ⁇ 0.01x10 -12 M, ⁇ 0.05x10 -12 M, or ⁇ 0.001x10-12M .
  • the K D value of the antigen-binding protein targeting GITR described in the present application for binding to the murine-derived GITR protein may be ⁇ 7x10-12M , ⁇ 6x10-12M , ⁇ 5x10-12M , ⁇ 4x10-12M , ⁇ 3x10 -12 M, ⁇ 2x10 -12 M, ⁇ 1x10 - 12 M, ⁇ 0.5x10 -12 M, ⁇ 0.1x10 -12 M, ⁇ 0.01x10 -12 M, ⁇ 0.05x10 -12 M, or ⁇ 0.001 x10-12M .
  • the K D value can also be determined by ELISA, competitive ELISA or BIACORE or KINEXA.
  • the antigen binding protein targeting GITR can stimulate the proliferation of immune cells.
  • the GITR-targeting antigen binding proteins described herein can maintain, enhance, accelerate or prolong immune cell proliferation, growth and/or survival in vivo or in vitro. Whether the GITR-targeting antigen binding proteins described herein can stimulate immunity can be determined using any method that can detect cell proliferation, growth and/or survival, such as cell proliferation assays or epithelial barrier integrity assays Cell Proliferation.
  • the immune cells may be selected from one or more of the group consisting of lymphocytes, natural killer cells and myeloid cells.
  • the lymphocytes can be B cells and/or T cells.
  • the myeloid cells may be selected from one or more of the group consisting of monocytes, macrophages, eosinophils, mast cells, basophils and granulocytes.
  • the antigen binding protein targeting GITR can stimulate immune cells to secrete IFN- ⁇ , and the secretion can be measured in T cell activity assay, for example, immune cells can be measured by ELISA, CBA or MSD method The content of secreted IFN- ⁇ .
  • the antigen binding protein targeting GITR can prevent, alleviate or treat cancer, for example, can reduce the tumor volume of GITR-positive cancer by at least about 10%, at least about 20%, at least about 30% , at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 99%, or 100%.
  • the number of tumor cells can be reduced by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about About 90%, at least about 99% or 100%.
  • the cancer may comprise a GITR positive cancer; and/or, the cancer comprises a PD-1 positive cancer.
  • the cancer may include solid tumors.
  • the cancer may be selected from the group consisting of colon cancer, rectal cancer and breast cancer.
  • the antigen binding protein targeting GITR can activate the GITR signaling pathway, so that the antigen binding protein targeting GITR described in the application can block the interaction between GITRL and its receptor GITR, and The functional response of T cells is restored from a dysfunctional state to an antigen-stimulated state.
  • the antigen-binding protein targeting GITR can inhibit the binding of GITR and GITRL, block the interaction between GITRL and its receptor GITR, and restore the functional response of T cells from a dysfunctional state to antigen stimulation state.
  • the immune checkpoint inhibitor can block the interaction of PD-1 and PD-L1.
  • the immune checkpoint inhibitor may comprise a PD-1 inhibitor.
  • the immune checkpoint inhibitor may comprise a PD-1 antibody or an antigen-binding fragment thereof.
  • the immune checkpoint inhibitor can comprise a mouse PD-1 antibody or an antigen-binding fragment thereof.
  • the immune checkpoint inhibitor can comprise a human PD-1 antibody or an antigen-binding fragment thereof.
  • the PD-1 antibody or its antigen-binding fragment can be selected from the following group: Pembrolizumab (Pembrolizumab, generic name Keytruda, K drug, manufactured by Merck), nivolumab (Opdivo, generic name Nivolumab, O medicine, manufactured by Bristol-Myers Squibb), Toripalizumab (manufactured by Junshi Bio), Sintilimab (manufactured by Innovent Bio), Camrelizumab (manufactured by Hengshi Bio) Rui Medicine) and Prelizumab (made by BeiGene).
  • the human PD-1 antibody or its antigen-binding fragment may comprise VL, and the VL may comprise the following amino acid sequences respectively:
  • the human PD-1 antibody or its antigen-binding fragment may comprise VH, and the VH may comprise the following amino acid sequences respectively:
  • the PD-1 antibody or antigen-binding fragment comprises HCDR3, wherein the HCDR3 of the PD-1 antibody or antigen-binding fragment may comprise the HCDR3 of RMP1-14.
  • RMP1-14 was purchased from Bioxcell, the product number is BE0146.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR2, wherein the HCDR2 of the PD-1 antibody or antigen-binding fragment may comprise the HCDR2 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR1, wherein the HCDR1 of the PD-1 antibody or antigen-binding fragment may comprise the HCDR1 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR3, wherein the LCDR3 of the PD-1 antibody or antigen-binding fragment may comprise the LCDR3 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR2, wherein the LCDR2 of the PD-1 antibody or antigen-binding fragment may comprise the LCDR2 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR1, wherein the LCDR1 of the PD-1 antibody or antigen-binding fragment may comprise the LCDR1 of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises VH, which may comprise the VH of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises VL, which may comprise the VL of RMP1-14.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR3, wherein the HCDR3 of the PD-1 antibody or antigen-binding fragment may comprise the HCDR3 of pembrolizumab; alternatively, may comprise the HCDR3 of nivolumab HCDR3.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR2, wherein the HCDR2 of the PD-1 antibody or antigen-binding fragment may comprise the HCDR2 of pembrolizumab; alternatively, may comprise the HCDR2 of nivolumab HCDR2.
  • the PD-1 antibody or antigen-binding fragment comprises HCDR1, wherein the HCDR1 of the PD-1 antibody or antigen-binding fragment may comprise the HCDR1 of pembrolizumab; alternatively, may comprise the HCDR1 of nivolumab HCDR1.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR3, wherein the LCDR3 of the PD-1 antibody or antigen-binding fragment may comprise the LCDR3 of pembrolizumab; or, may comprise the LCDR3 of nivolumab LCDR3.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR2, wherein the LCDR2 of the PD-1 antibody or antigen-binding fragment may comprise the LCDR2 of pembrolizumab; or, may comprise the LCDR2 of nivolumab LCDR2.
  • the PD-1 antibody or antigen-binding fragment comprises LCDR1, wherein the LCDR1 of the PD-1 antibody or antigen-binding fragment may comprise LCDR1 of pembrolizumab; alternatively, may comprise nivolumab Anti-LCDR1.
  • the PD-1 antibody or antigen-binding fragment comprises a VH, which may comprise the VH of pembrolizumab; alternatively, may comprise the VH of nivolumab.
  • the PD-1 antibody or antigen-binding fragment comprises a VL, which may comprise the VL of pembrolizumab; alternatively, may comprise the VL of nivolumab.
  • the present application provides a pharmaceutical product comprising the GITR-targeting antigen binding protein described herein and the immune checkpoint inhibitor described herein.
  • the pharmaceutical product may comprise a pharmaceutical composition.
  • the pharmaceutical composition may also include a pharmaceutically acceptable agent.
  • the pharmaceutically acceptable agent may include stabilizers, pH adjusters and/or excipients.
  • the pharmaceutical composition may have a dosage form.
  • the dosage form may comprise a liquid formulation.
  • the GITR-targeting antigen binding protein and the immune checkpoint inhibitor in the drug product may not be mixed with each other.
  • the GITR-targeting antigen binding protein and the immune checkpoint inhibitor in the drug product may be present in separate containers.
  • the container may comprise a glass vial for injection.
  • the application provides the use of the pharmaceutical product described in the application in the preparation of a medicament for preventing, relieving or treating cancer.
  • the present application provides the use of the antigen binding protein targeting GITR and the immune checkpoint inhibitor described in the present application in the preparation of a medicament for preventing, relieving or treating cancer.
  • the present application provides a method of preventing, relieving or treating cancer, comprising the steps of: administering the GITR-targeting antigen binding protein described in the present application to a subject in need, and/or, to a subject in need of subjects administered the immune checkpoint inhibitors described herein.
  • the method may comprise the step of administering the pharmaceutical product described herein to a subject in need thereof.
  • the present application provides the GITR-targeting antigen binding protein described herein, and/or the pharmaceutical product described herein, for use in preventing, alleviating or treating cancer.
  • the cancer may comprise a GITR positive cancer; and/or the cancer may comprise a PD-1 positive cancer.
  • the cancer may include solid tumors.
  • the cancer may be selected from the group consisting of colon cancer, rectal cancer and breast cancer.
  • the administration dose of the antigen binding protein targeting GITR may be about 1 mg/Kg to about 10 mg/Kg, for example, it may be about 1 mg/Kg to about 9 mg/Kg, about 1 mg/Kg to about 8mg/Kg, about 1mg/Kg-about 7mg/Kg, about 1mg/Kg-about 6mg/Kg, about 1mg/Kg-about 5mg/Kg, about 1mg/Kg-about 3mg/Kg, about 3mg/Kg-about 6 mg/Kg or about 1 mg/Kg to about 3 mg/Kg; for example, can be about 1 mg/Kg, about 2 mg/Kg, about 3 mg/Kg, about 4 mg/Kg, about 5 mg/Kg, about 6 mg/Kg, about 7 mg /Kg, about 8 mg/Kg, about 9 mg/Kg, or about 10 mg/Kg.
  • the administration dose of the immune checkpoint inhibitor can be about 1 mg/Kg-about 10 mg/Kg, for example, it can be about 1 mg/Kg-about 9 mg/Kg, about 1 mg/Kg-about 8 mg/Kg/ Kg, about 1mg/Kg-about 7mg/Kg, about 1mg/Kg-about 6mg/Kg, about 1mg/Kg-about 5mg/Kg, about 1mg/Kg-about 3mg/Kg, about 3mg/Kg-about 6mg/Kg Kg or about 1 mg/Kg to about 3 mg/Kg; for example, can be about 1 mg/Kg, about 2 mg/Kg, about 3 mg/Kg, about 4 mg/Kg, about 5 mg/Kg, about 6 mg/Kg, about 7 mg/Kg , about 8 mg/Kg, about 9 mg/Kg, or about 10 mg/Kg.
  • the administration frequency of the antigen binding protein targeting GITR may be twice a week.
  • the administration frequency of the immune checkpoint inhibitor may be twice a week.
  • the administration mode of the antigen binding protein targeting GITR can be injection.
  • it can be intraperitoneal, intramuscular or intravenous.
  • the administration mode of the immune checkpoint inhibitor can be injection.
  • it can be intraperitoneal, intramuscular or intravenous.
  • the concentration of the antigen binding protein targeting GITR may be about 1 mg/mL to about 10 mg/mL, for example, it may be about 1 mg/mL to about 9 mg/mL, about 1 mg/mL to about 8 mg/mL mL, about 1 mg/mL to about 7 mg/mL, about 1 mg/mL to about 6 mg/mL, about 1 mg/mL to about 5 mg/mL, about 1 mg/mL to about 4 mg/mL, about 1 mg/mL to about 3 mg/mL mL or about 1 mg/mL to about 2 mg/mL; for example, it can be about 1 mg/mL, about 1.5 mg/mL, about 2 mg/mL, about 2.5 mg/mL, about 2.8 mg/mL, about 3 mg/mL, about 3.5 mg/mL mg/mL, about 4 mg/mL, about 4.5 mg/mL, or about 5 mg/mL.
  • the concentration of the immune checkpoint inhibitor may be about 1 mg/mL to about 10 mg/mL, for example, it may be about 1 mg/mL to about 9 mg/mL, about 1 mg/mL to about 8 mg/mL, about 1 mg/mL to about 7 mg/mL, about 1 mg/mL to about 6 mg/mL, about 1 mg/mL to about 5 mg/mL, about 1 mg/mL to about 4 mg/mL, about 1 mg/mL to about 3 mg/mL or About 1 mg/mL to about 2 mg/mL; for example can be about 1 mg/mL, about 1.5 mg/mL, about 2 mg/mL, about 2.5 mg/mL, about 3 mg/mL, about 3.5 mg/mL, about 4 mg/mL , about 4.5 mg/mL or about 5 mg/mL.
  • the antibodies were humanized by changing certain amino acid residues in the framework regions of the variable regions of the heavy and light chains, and a total of 9 humanized antibodies shown in Table 1 were obtained, that is, 9 isolated antigens described in this application Binding proteins, represented by 3E2 1-2, 3E2 1-3, 3E2 1-4, 3E2 2-2, 3E2 2-3, 3E2 2-4, 3E2 3-2, 3E2 3-3, and 3E2 3-4, respectively .
  • the humanized VH gene and VL gene described above were synthesized, and the VH and human IgG1 heavy chain constant regions constituted the heavy chain of the antibody, and the VL and human kappa light chain constant regions constituted the light chain of the antibody.
  • Each gene was cloned into the pcDNA4/myc-HisA vector to obtain a heavy chain expression plasmid and a light chain expression plasmid.
  • the heavy chain expression plasmid and the light chain expression plasmid were paired according to Table 1 and then transiently transfected into HEK293 (ATCC, CRL-1573 TM ) cell line for protein production.
  • Example 2 The drug product described in this application inhibits tumor growth
  • C57-GITR KI mice that is, C57 background mice with gene editing knock-in human GITR, purchased from Biocytometer
  • C57-GITR KI mice were subcutaneously inoculated with 100 ⁇ L of MC38 cell suspension (where the cell suspension concentration was 5 ⁇ 10 6 cells/mL, the cell viability in the cell suspension was 85.87%) to obtain a mouse tumor model.
  • mice PD1 antibody is RMP1-14, which was purchased from Bioxcell, the product number is BE0146.
  • Figures 2A-2D show tumors in each mouse in the human IgG1-treated group, the 3E2 1-4-treated group, the mouse PD1 antibody-treated group, and the 3E2 1-4+mouse PD1 antibody-treated group (ie, treatment groups 1-4), respectively. Volume statistics. The efficacy analysis of each treatment group is shown in Table 3.
  • Example 3 Antigen-binding protein targeting GITR described in this application inhibits tumor growth
  • C57-GITR KI mice that is, C57 background mice with gene editing knock-in human GITR, purchased from Bio-Ocelot
  • 100 ⁇ L of MC38 cell suspension was subcutaneously inoculated into C57-GITR KI mice to obtain a mouse tumor model.
  • mice tumor models were administered according to the manner in Table 4.
  • mice The body weight and tumor volume of the mice in each treatment group were measured during the administration period, and the results are shown in Figure 3A and Figure 3B, respectively.
  • Figures 4A-4D show the human IgG1 treatment group, the 1 mg/Kg 3E2 1-4 treatment group, the 3 mg/Kg 3E2 1-4 treatment group, and the 10 mg/Kg 3E2 1-4 treatment group (ie, treatment groups 1-4), respectively.
  • the efficacy analysis of each treatment group is shown in Table 5.
  • Example 4 The drug product described in this application inhibits tumor growth
  • C57-GITR KI mice that is, C57 background mice with gene editing knock-in human GITR, purchased from Biocytometer
  • C57-GITR KI mice were subcutaneously inoculated with 100 ⁇ L of MC38 cell suspension (where the cell suspension concentration was 5 ⁇ 10 6 cells/mL, the cell viability in the cell suspension was 85.87%) to obtain a mouse tumor model.
  • mice PD1 antibody is RMP1-14, which was purchased from Bioxcell, the product number is BE0146.
  • mice The body weight and tumor volume of the mice in each treatment group were measured during the administration period, and the results are shown in Figure 5A and Figure 5B, respectively.
  • Figure 5 shows the tumor volume of each mouse in the human IgG1 treatment group, the 3E2 1-4 treatment group, the mouse PD1 antibody treatment group, and the 3E2 1-4+ mouse PD1 antibody treatment group (ie, treatment groups 1-4), respectively. statistical results.

Abstract

L'invention concerne un produit pharmaceutique, comprenant une protéine de liaison à l'antigène ciblant GITR et un inhibiteur de point de contrôle immunitaire, la protéine de liaison à l'antigène comprenant au moins une CDR dans VH ayant une séquence d'acides aminés telle que représentée dans SEQ ID NO : 23, et comprend au moins un CDR dans VL ayant une séquence d'acides aminés telle que représentée dans SEQ ID NO : 24. L'invention concerne également une utilisation du produit pharmaceutique dans le traitement du cancer.
PCT/CN2021/142299 2021-01-08 2021-12-29 Produit pharmaceutique et son utilisation WO2022148279A1 (fr)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2772613A1 (fr) * 2009-09-03 2011-03-10 Schering Corporation Anticorps anti-gitr
WO2015031667A2 (fr) * 2013-08-30 2015-03-05 Amgen Inc. Protéines de liaison à l'antigène gitr
US20150353637A1 (en) * 2014-06-06 2015-12-10 Bristol-Myers Squibb Company Antibodies against glucocorticoid-induced tumor necrosis factor receptor (gitr) and uses thereof
CN105492463A (zh) * 2013-08-20 2016-04-13 默沙东公司 肿瘤免疫调节
US20170260282A1 (en) * 2016-03-08 2017-09-14 Janssen Biotech, Inc. GITR Antibodies, Methods, and Uses
US20170355774A1 (en) * 2016-06-10 2017-12-14 Regeneron Pharmaceuticals, Inc. Anti-gitr antibodies and uses thereof
US20190364861A1 (en) * 2017-06-09 2019-12-05 Beijing Biocytogen Co., Ltd. Genetically modified non-human animal with human or chimeric gitr

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2772613A1 (fr) * 2009-09-03 2011-03-10 Schering Corporation Anticorps anti-gitr
CN105492463A (zh) * 2013-08-20 2016-04-13 默沙东公司 肿瘤免疫调节
WO2015031667A2 (fr) * 2013-08-30 2015-03-05 Amgen Inc. Protéines de liaison à l'antigène gitr
US20150353637A1 (en) * 2014-06-06 2015-12-10 Bristol-Myers Squibb Company Antibodies against glucocorticoid-induced tumor necrosis factor receptor (gitr) and uses thereof
US20170260282A1 (en) * 2016-03-08 2017-09-14 Janssen Biotech, Inc. GITR Antibodies, Methods, and Uses
US20170355774A1 (en) * 2016-06-10 2017-12-14 Regeneron Pharmaceuticals, Inc. Anti-gitr antibodies and uses thereof
US20190364861A1 (en) * 2017-06-09 2019-12-05 Beijing Biocytogen Co., Ltd. Genetically modified non-human animal with human or chimeric gitr

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