WO2022148279A1 - Pharmaceutical product and use thereof - Google Patents

Pharmaceutical product and use thereof 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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French (fr)
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/en
Publication of WO2022148279A1 publication Critical patent/WO2022148279A1/en

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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

A pharmaceutical product, comprising an antigen-binding protein targeting GITR and an immune checkpoint inhibitor, wherein the antigen-binding protein comprises at least one CDR in the VH having an amino acid sequence as shown in SEQ ID NO: 23, and comprises at least one CDR in the VL having an amino acid sequence as shown in SEQ ID NO: 24. Also provided is a use of the pharmaceutical product in the treatment of cancer.

Description

药物产品及其用途Pharmaceutical products and their uses 技术领域technical field
本申请涉及生物医药领域，具体的涉及一种药物产品及其用途。The present application relates to the field of biomedicine, in particular to a pharmaceutical product and use thereof.
背景技术Background technique
GITR是TNF受体家族的一个成员，GITR是分子量大小约为26kDa的I型跨膜蛋白，和TNF受体家族其它分子有14-28％的同源性(Gurney AL et al，1999)。GITR组成型在Treg上高表达，在未活化的CD4 +T细胞、CD8 +T细胞、NK和NKT细胞上低表达。当T细胞活化后，GITR在CD4 +T、CD8 +T和NK细胞高表达升高(S.Ronchetti，et al.,2004)。GITR天然配体GITRL在APC上表达，GITRL结合GITR后，提供共刺激信号，调节抗原特异性T细胞应答，加强细胞免疫和体液免疫(Selvakumar Sukumar et al.，2017)。 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).
发明内容SUMMARY OF THE INVENTION
本申请提供了一种药物产品，其包含靶向GITR的抗原结合蛋白和免疫检查点抑制剂。所述药物产品具备下组中一个或多个有益效果：1)有效地预防、缓解或治疗癌症；2)所述靶向GITR的抗原结合蛋白和所述免疫检查点抑制剂存在协同作用。本申请还提供了所述药物产品在预防、缓解或治疗癌症，尤其是GITR阳性癌症和/或PD-1阳性癌症中的用途和制药用途。本申请还提供了所述靶向GITR的抗原结合蛋白在预防、缓解或治疗癌症，尤其是GITR阳性癌症的用途。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.
一方面，本申请提供了一种药物产品，其包含靶向GITR的抗原结合蛋白和免疫检查点抑制剂，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中的至少一个CDR，且包含氨基酸序列如SEQ ID NO:24所示的VL中的至少一个CDR。In one aspect, 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.
在某些实施方式中，所述药物产品为药物组合物。In certain embodiments, the pharmaceutical product is a pharmaceutical composition.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR3。In certain embodiments, the antigen binding protein comprises HCDR3 in VH having the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR2。In certain embodiments, the antigen binding protein comprises HCDR2 in VH having the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR1。In certain embodiments, the antigen binding protein comprises HCDR1 in VH having the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR1。In certain embodiments, the antigen binding protein comprises LCDR1 in VL having the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR2。In certain embodiments, the antigen binding protein comprises LCDR2 in VL having the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR3。In certain embodiments, the antigen binding protein comprises LCDR3 in VL having the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述抗原结合蛋白包含HCDR3，所述HCDR3包含SEQ ID NO:2所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:2.
在某些实施方式中，所述抗原结合蛋白包含HCDR2，所述HCDR2包含SEQ ID NO:4所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:4.
在某些实施方式中，所述抗原结合蛋白包含HCDR1，所述HCDR1包含SEQ ID NO:3所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:3.
在某些实施方式中，所述抗原结合蛋白包含LCDR3，所述LCDR3包含SEQ ID NO:12所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:12.
在某些实施方式中，所述抗原结合蛋白包含LCDR2，所述LCDR2包含SEQ ID NO:11所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:11.
在某些实施方式中，所述抗原结合蛋白包含LCDR1，所述LCDR1包含SEQ ID NO:10所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:10.
在某些实施方式中，所述抗原结合蛋白包括抗体或其抗原结合片段。In certain embodiments, the antigen-binding protein comprises an antibody or antigen-binding fragment thereof.
在某些实施方式中，所述抗原结合片段包括Fab，Fab’，F(ab)2、Fv片段、F(ab’)2，scFv，di-scFv和/或dAb。In certain embodiments, the antigen-binding fragment comprises Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
在某些实施方式中，所述抗体为人源化抗体。In certain embodiments, the antibody is a humanized antibody.
在某些实施方式中，所述VL包括框架区L-FR1，L-FR2，L-FR3，和L-FR4。In certain embodiments, the VL includes framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
在某些实施方式中，所述L-FR1的C末端与所述LCDR1的N末端直接或间接相连，且所述L-FR1包含SEQ ID NO:39所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR1包含SEQ ID NO:13、30、34中任一项所示的氨基酸序列。In certain embodiments, the L-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 13, 30, 34.
在某些实施方式中，所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包 含SEQ ID NO:40所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR2包含SEQ ID NO:14、31中任一项所示的氨基酸序列。In certain embodiments, the L-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 14, 31.
在某些实施方式中，所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO:41所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR3包含SEQ ID NO:15、32中任一项所示的氨基酸序列。In certain embodiments, the L-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 15, 32.
在某些实施方式中，所述L-FR4的N末端与所述LCDR3的C末端相连，且所述L-FR4包含SEQ ID NO:42所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR4包含SEQ ID NO:16、33中任一项所示的氨基酸序列。In certain embodiments, the L-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 16, 33.
在某些实施方式中，所述VL包含SEQ ID NO:24所示的氨基酸序列。In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述VL包含SEQ ID NO:9、20-22中任一项所示的氨基酸序列。In certain embodiments, the VL comprises the amino acid sequence set forth in any one of SEQ ID NOs: 9, 20-22.
在某些实施方式中，所述抗原结合蛋白包括抗体轻链恒定区，且所述抗体轻链恒定区包括人Igκ恒定区。In certain embodiments, the antigen binding protein comprises an antibody light chain constant region, and the antibody light chain constant region comprises a human IgK constant region.
在某些实施方式中，所述抗体轻链恒定区包含SEQ ID NO:52所示的氨基酸序列。In certain embodiments, the antibody light chain constant region comprises the amino acid sequence set forth in SEQ ID NO:52.
在某些实施方式中，所述抗原结合蛋白包含抗体轻链LC，且所述LC包含SEQ ID NO:50、46-48中任一项所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises an antibody light chain LC, and the LC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 50, 46-48.
在某些实施方式中，所述VH包括框架区H-FR1，H-FR2，H-FR3，和H-FR4。In certain embodiments, the VH includes framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
在某些实施方式中，所述H-FR1的C末端与所述HCDR1的N末端直接或间接相连，且所述H-FR1包含SEQ ID NO:35所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR1包含SEQ ID NO:5、25、29中任一项所示的氨基酸序列。In certain embodiments, the H-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 5, 25, 29.
在某些实施方式中，所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO:36所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR2包含SEQ ID NO:6、26中任一项所示的氨基酸序列。In certain embodiments, the H-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 6, 26.
在某些实施方式中，所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO:37所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR3包含SEQ ID NO:7、27中任一项所示的氨基酸序列。In certain embodiments, the H-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 7, 27.
在某些实施方式中，所述H-FR4的N末端与所述HCDR3的C末端相连，且所述H-FR4包含SEQ ID NO:38所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR4包含SEQ ID NO:8、28中任一项所示的氨基酸序列。In certain embodiments, the H-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 8, 28.
在某些实施方式中，所述VH包含SEQ ID NO:23所示的氨基酸序列。In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述VH包含SEQ ID NO:1、17-19中任一项所示的氨基酸序列。In certain embodiments, the VH comprises the amino acid sequence set forth in any one of SEQ ID NOs: 1, 17-19.
在某些实施方式中，所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG重链恒定区。In certain embodiments, 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.
在某些实施方式中，所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG1重链恒定区。In certain embodiments, 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.
在某些实施方式中，所述抗体重链恒定区包含SEQ ID NO:51所示的氨基酸序列。In certain embodiments, the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:51.
在某些实施方式中，所述抗原结合蛋白包含抗体重链HC，且所述HC包含SEQ ID NO:49、43-45中任一项所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises an antibody heavy chain HC, and the HC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 49, 43-45.
在某些实施方式中，所述免疫检查点抑制剂阻碍PD-1和PD-L1的相互作用。In certain embodiments, the immune checkpoint inhibitor blocks the interaction of PD-1 and PD-L1.
在某些实施方式中，所述免疫检查点抑制剂包含PD-1抑制剂。In certain embodiments, the immune checkpoint inhibitor comprises a PD-1 inhibitor.
在某些实施方式中，所述免疫检查点抑制剂包含PD-1抗体或其抗原结合片段。In certain embodiments, the immune checkpoint inhibitor comprises a PD-1 antibody or antigen-binding fragment thereof.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含HCDR3，其中所述PD-1抗体或抗原结合片段的HCDR3包含RMP1-14的HCDR3。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含HCDR2，其中所述PD-1抗体或抗原结合片段的HCDR2包含RMP1-14的HCDR2。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含HCDR1，其中所述PD-1抗体或抗原结合片段的HCDR1包含RMP1-14的HCDR1。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含LCDR3，其中所述PD-1抗体或抗原结合片段的LCDR3包含RMP1-14的LCDR3。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含LCDR2，其中所述PD-1抗体或抗原结合片段的LCDR2包含RMP1-14的LCDR2。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含LCDR1，其中所述PD-1抗体或抗原结合片段的所述LCDR1包含RMP1-14的LCDR1。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含VH，所述VH包含RMP1-14的VH。In certain embodiments, the PD-1 antibody or antigen-binding fragment comprises a VH comprising the VH of RMP1-14.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含VL，所述VL包含RMP1-14的VL。In certain embodiments, the PD-1 antibody or antigen-binding fragment comprises a VL comprising the VL of RMP1-14.
在某些实施方式中，所述靶向GITR的抗原结合蛋白和所述免疫检查点抑制剂在所述药物产品中相互不混合。In certain embodiments, the GITR-targeting antigen binding protein and the immune checkpoint inhibitor are immiscible with each other in the drug product.
在某些实施方式中，所述靶向GITR的抗原结合蛋白和所述免疫检查点抑制剂在所述药物产品中分别存在于不同的容器中。In certain embodiments, the GITR-targeting antigen binding protein and the immune checkpoint inhibitor are present in separate containers in the drug product.
另一方面，本申请提供本申请所述的药物产品在制备预防、缓解或治疗癌症的药物中的用途。In another aspect, 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.
在某些实施方式中，所述癌症包括GITR阳性的癌症；和/或，所述癌症包括PD-1阳性的癌症。In certain embodiments, the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
在某些实施方式中，所述癌症包括实体瘤。In certain embodiments, the cancer comprises a solid tumor.
在某些实施方式中，所述癌症选自下组：结肠癌、直肠癌和乳腺癌。In certain embodiments, the cancer is selected from the group consisting of colon cancer, rectal cancer, and breast cancer.
另一方面，本申请提供靶向GITR的抗原结合蛋白和免疫检查点拮抗剂在制备预防、缓解或治疗癌症的药物中的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中的至少一个CDR，且包含氨基酸序列如SEQ ID NO:24所示的VL中的至少一个CDR。In another aspect, 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.
在某些实施方式中，所述癌症包括GITR阳性的癌症；和/或，所述癌症包括PD-1阳性的癌症。In certain embodiments, the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
在某些实施方式中，所述癌症包括实体瘤。In certain embodiments, the cancer comprises a solid tumor.
在某些实施方式中，所述癌症选自下组：结肠癌、直肠癌和乳腺癌。In certain embodiments, the cancer is selected from the group consisting of colon cancer, rectal cancer, and breast cancer.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR3。In certain embodiments, the antigen binding protein comprises HCDR3 in VH having the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR2。In certain embodiments, the antigen binding protein comprises HCDR2 in VH having the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR1。In certain embodiments, the antigen binding protein comprises HCDR1 in VH having the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR1。In certain embodiments, the antigen binding protein comprises LCDR1 in VL having the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR2。In certain embodiments, the antigen binding protein comprises LCDR2 in VL having the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR3。In certain embodiments, the antigen binding protein comprises LCDR3 in VL having the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述抗原结合蛋白包含HCDR3，所述HCDR3包含SEQ ID NO:2所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:2.
在某些实施方式中，所述抗原结合蛋白包含HCDR2，所述HCDR2包含SEQ ID NO:4所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:4.
在某些实施方式中，所述抗原结合蛋白包含HCDR1，所述HCDR1包含SEQ ID NO:3所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:3.
在某些实施方式中，所述抗原结合蛋白包含LCDR3，所述LCDR3包含SEQ ID NO:12所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:12.
在某些实施方式中，所述抗原结合蛋白包含LCDR2，所述LCDR2包含SEQ ID NO:11所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:11.
在某些实施方式中，所述抗原结合蛋白包含LCDR1，所述LCDR1包含SEQ ID NO:10所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:10.
在某些实施方式中，所述抗原结合蛋白包括抗体或其抗原结合片段。In certain embodiments, the antigen-binding protein comprises an antibody or antigen-binding fragment thereof.
在某些实施方式中，所述抗原结合片段包括Fab，Fab’，F(ab)2、Fv片段、F(ab’)2，scFv，di-scFv和/或dAb。In certain embodiments, the antigen-binding fragment comprises Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
在某些实施方式中，所述抗体为人源化抗体。In certain embodiments, the antibody is a humanized antibody.
在某些实施方式中，所述VL包括框架区L-FR1，L-FR2，L-FR3，和L-FR4。In certain embodiments, the VL includes framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
在某些实施方式中，所述L-FR1的C末端与所述LCDR1的N末端直接或间接相连，且所述L-FR1包含SEQ ID NO:39所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR1包含SEQ ID NO:13、30、34中任一项所示的氨基酸序列。In certain embodiments, the L-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 13, 30, 34.
在某些实施方式中，所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包含SEQ ID NO:40所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR2包含SEQ ID NO:14、31中任一项所示的氨基酸序列。In certain embodiments, the L-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 14, 31.
在某些实施方式中，所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO:41所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR3包含SEQ ID NO:15、32中任一项所示的氨基酸序列。In certain embodiments, the L-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 15, 32.
在某些实施方式中，所述L-FR4的N末端与所述LCDR3的C末端相连，且所述L-FR4包含SEQ ID NO:42所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述L-FR4包含SEQ ID NO:16、33中任一项所示的氨基酸序列。In certain embodiments, the L-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 16, 33.
在某些实施方式中，所述VL包含SEQ ID NO:24所示的氨基酸序列。In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:24.
在某些实施方式中，所述VL包含SEQ ID NO:9、20-22中任一项所示的氨基酸序列。In certain embodiments, the VL comprises the amino acid sequence set forth in any one of SEQ ID NOs: 9, 20-22.
在某些实施方式中，所述抗原结合蛋白包括抗体轻链恒定区，且所述抗体轻链恒定区包括人Igκ恒定区。In certain embodiments, the antigen binding protein comprises an antibody light chain constant region, and the antibody light chain constant region comprises a human IgK constant region.
在某些实施方式中，所述抗体轻链恒定区包含SEQ ID NO:52所示的氨基酸序列。In certain embodiments, the antibody light chain constant region comprises the amino acid sequence set forth in SEQ ID NO:52.
在某些实施方式中，所述抗原结合蛋白包含抗体轻链LC，且所述LC包含SEQ ID NO:50、46-48中任一项所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises an antibody light chain LC, and the LC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 50, 46-48.
在某些实施方式中，所述VH包括框架区H-FR1，H-FR2，H-FR3，和H-FR4。In certain embodiments, the VH includes framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
在某些实施方式中，所述H-FR1的C末端与所述HCDR1的N末端直接或间接相连，且所述H-FR1包含SEQ ID NO:35所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR1包含SEQ ID NO:5、25、29中任一项所示的氨基酸序列。In certain embodiments, the H-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 5, 25, 29.
在某些实施方式中，所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO:36所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR2包含SEQ ID NO:6、26中任一项所示的氨基酸序列。In certain embodiments, the H-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 6, 26.
在某些实施方式中，所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO:37所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR3包含SEQ ID NO:7、27中任一项所示的氨基酸序列。In certain embodiments, the H-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 7, 27.
在某些实施方式中，所述H-FR4的N末端与所述HCDR3的C末端相连，且所述H-FR4包含SEQ ID NO:38所示的氨基酸序列。In certain embodiments, 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.
在某些实施方式中，所述H-FR4包含SEQ ID NO:8、28中任一项所示的氨基酸序列。In certain embodiments, the H-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 8, 28.
在某些实施方式中，所述VH包含SEQ ID NO:23所示的氨基酸序列。In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO:23.
在某些实施方式中，所述VH包含SEQ ID NO:1、17-19中任一项所示的氨基酸序列。In certain embodiments, the VH comprises the amino acid sequence set forth in any one of SEQ ID NOs: 1, 17-19.
在某些实施方式中，所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG重链恒定区。In certain embodiments, 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.
在某些实施方式中，所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG1重链恒定区。In certain embodiments, 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.
在某些实施方式中，所述抗体重链恒定区包含SEQ ID NO:51所示的氨基酸序列。In certain embodiments, the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:51.
在某些实施方式中，所述抗原结合蛋白包含抗体重链HC，且所述HC包含SEQ ID NO:49、43-45中任一项所示的氨基酸序列。In certain embodiments, the antigen binding protein comprises an antibody heavy chain HC, and the HC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 49, 43-45.
在某些实施方式中，所述免疫检查点抑制剂阻碍PD-1和PD-L1的相互作用。In certain embodiments, the immune checkpoint inhibitor blocks the interaction of PD-1 and PD-L1.
在某些实施方式中，所述免疫检查点抑制剂包含PD-1抑制剂。In certain embodiments, the immune checkpoint inhibitor comprises a PD-1 inhibitor.
在某些实施方式中，所述免疫检查点抑制剂包含PD-1抗体或其抗原结合片段。In certain embodiments, the immune checkpoint inhibitor comprises a PD-1 antibody or antigen-binding fragment thereof.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含HCDR3，其中所述PD-1抗体或抗原结合片段的HCDR3包含RMP1-14的HCDR3。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含HCDR2，其中所述PD-1抗体或抗原结合片段的HCDR2包含RMP1-14的HCDR2。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含HCDR1，其中所述PD-1抗体或抗原结合片段的HCDR1包含RMP1-14的HCDR1。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含LCDR3，其中所述PD-1抗体或抗原结合片段的LCDR3包含RMP1-14的LCDR3。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含LCDR2，其中所述PD-1抗体或抗原结合片段的LCDR2包含RMP1-14的LCDR2。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含LCDR1，其中所述PD-1抗体或抗原结合片段的所述LCDR1包含RMP1-14的LCDR1。In certain embodiments, 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.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含VH，所述VH包含RMP1-14的VH。In certain embodiments, the PD-1 antibody or antigen-binding fragment comprises a VH comprising the VH of RMP1-14.
在某些实施方式中，所述PD-1抗体或抗原结合片段包含VL，所述VL包含RMP1-14的VL。In certain embodiments, the PD-1 antibody or antigen-binding fragment comprises a VL comprising the VL of RMP1-14.
另一方面，本申请提供一种预防、缓解或治疗癌症的方法，其包括以下步骤：向有需要的受试者施用本申请所述的抗原结合蛋白，和/或，向有需要的受试者施用本申请所述的免疫检查点抑制剂。In another aspect, 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.
在某些实施方式中，所述方法包括以下步骤：向有需要的受试者施用本申请所述的药物产品。In certain embodiments, the method comprises the step of administering to a subject in need thereof a pharmaceutical product described herein.
在某些实施方式中，所述癌症包括GITR阳性的癌症；和/或，所述癌症包括PD-1阳性的癌症。In certain embodiments, the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
在某些实施方式中，所述癌症包括实体瘤。In certain embodiments, the cancer comprises a solid tumor.
在某些实施方式中，所述癌症选自下组：结肠癌、直肠癌和乳腺癌。In certain embodiments, the cancer is selected from the group consisting of colon cancer, rectal cancer, and breast cancer.
在某些实施方式中，所述抗原结合蛋白的给药剂量为1mg/Kg-10mg/Kg。In certain embodiments, the antigen binding protein is administered at a dose of 1 mg/Kg to 10 mg/Kg.
在某些实施方式中，所述免疫检查点抑制剂的给药剂量为1mg/Kg-10mg/Kg。In certain embodiments, the immune checkpoint inhibitor is administered at a dose of 1 mg/Kg to 10 mg/Kg.
在某些实施方式中，所述抗原结合蛋白的给药频率为每周2次。In certain embodiments, the frequency of administration of the antigen binding protein is twice a week.
在某些实施方式中，所述免疫检查点抑制剂的给药频率为每周2次。In certain embodiments, the frequency of administration of the immune checkpoint inhibitor is twice a week.
在某些实施方式中，所述抗原结合蛋白的给药方式为注射。In certain embodiments, the antigen binding protein is administered by injection.
在某些实施方式中，所述免疫检查点抑制剂的给药方式为注射。In certain embodiments, the immune checkpoint inhibitor is administered by injection.
在某些实施方式中，所述抗原结合蛋白的浓度为1mg/mL-10mg/mL。In certain embodiments, the antigen binding protein is at a concentration of 1 mg/mL to 10 mg/mL.
在某些实施方式中，所述免疫检查点抑制剂的浓度为1mg/mL-10mg/mL。In certain embodiments, the concentration of the immune checkpoint inhibitor is 1 mg/mL to 10 mg/mL.
本领域技术人员能够从下文的详细描述中容易地洞察到本申请的其它方面和优势。下文的详细描述中仅显示和描述了本申请的示例性实施方式。如本领域技术人员将认识到的，本申请的内容使得本领域技术人员能够对所公开的具体实施方式进行改动而不脱离本申请所涉及发明的精神和范围。相应地，本申请的附图和说明书中的描述仅仅是示例性的，而非为限制性的。Other aspects and advantages of the present application can be readily appreciated by those skilled in the art from the following detailed description. Only exemplary embodiments of the present application are shown and described in the following detailed description. As those skilled in the art will recognize, the content of this application enables those skilled in the art to make changes to the specific embodiments disclosed without departing from the spirit and scope of the invention to which this application relates. Accordingly, the drawings and descriptions in the specification of the present application are only exemplary and not restrictive.
附图说明Description of drawings
本申请所涉及的发明的具体特征如所附权利要求书所显示。通过参考下文中详细描述的示例性实施方式和附图能够更好地理解本申请所涉及发明的特点和优势。对附图简要说明书如下：The invention to which this application relates is set forth with particularity characteristic of the appended claims. The features and advantages of the inventions involved in this application can be better understood by reference to the exemplary embodiments described in detail hereinafter and the accompanying drawings. A brief description of the drawings is as follows:
图1A显示的是本申请所述药物产品对小鼠体重的影响。Figure 1A shows the effect of the drug product described in this application on the body weight of mice.
图1B显示的是本申请所述药物产品对小鼠肿瘤体积的影响。Figure 1B shows the effect of the drug product described herein on tumor volume in mice.
图2A-2D显示了不同的治疗组对小鼠肿瘤体积的影响。Figures 2A-2D show the effect of different treatment groups on tumor volume in mice.
图3A显示的是本申请所述靶向GITR的抗原结合蛋白对小鼠体重的影响。Figure 3A shows the effect of the GITR-targeting antigen binding proteins described herein on the body weight of mice.
图3B显示的是本申请靶向GITR的抗原结合蛋白对小鼠肿瘤体积的影响。Figure 3B shows the effect of the antigen-binding protein targeting GITR of the present application on tumor volume in mice.
图4A-4D显示了不同的治疗组对小鼠肿瘤体积的影响。Figures 4A-4D show the effect of different treatment groups on tumor volume in mice.
图5A-5B显示了本申请所述药物产品对小鼠体重和小鼠肿瘤体积的影响。Figures 5A-5B show the effect of the drug products described herein on mouse body weight and mouse tumor volume.
具体实施方式Detailed ways
以下由特定的具体实施例说明本申请发明的实施方式，熟悉此技术的人士可由本说明书所公开的内容容易地了解本申请发明的其他优点及效果。The embodiments of the invention of the present application are described below with specific specific examples, and those skilled in the art can easily understand other advantages and effects of the invention of the present application from the contents disclosed in this specification.
术语定义Definition of Terms
以下对本申请做进一步描述：在本申请中，除非另有说明，否则使用的科学和技术名词具有本领域技术人员所通常理解的含义。并且，所用的蛋白质和核酸化学、分子生物学、细胞和组织培养、微生物学、免疫学相关术语和实验室操作步骤均为相应领域内广泛使用的术语和常规步骤。同时，为了更好地理解本申请，下面提供相关术语的定义和解释。The application is further described below: In this application, unless otherwise specified, the scientific and technical terms used have the meanings commonly understood by those skilled in the art. In addition, the terms and laboratory procedures used in protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, immunology and laboratory procedures are the terms and routine procedures widely used in the corresponding fields. Meanwhile, for a better understanding of the present application, definitions and explanations of related terms are provided below.
在本申请中，术语“抗原结合蛋白”通常是指包含结合抗原的部分的蛋白质，以及任选地 允许结合抗原的部分采用促进抗原结合蛋白与抗原结合的构象的支架或骨架部分。抗原结合蛋白的实例包括但不限于抗体、抗原结合片段(Fab，Fab’，F(ab) 2，Fv片段，F(ab’) 2，scFv，di-scFv和/或dAb)、免疫缀合物、多特异性抗体(例如双特异性抗体)、抗体片段、抗体衍生物、抗体类似物或融合蛋白等，只要它们显示出所需的抗原结合活性即可。在本申请中，所述“分离的抗原结合蛋白”可以包含结合抗原的部分和任选地，允许抗原结合部分采用促进所述抗原结合部分结合抗原的构象的支架或构架部分。例如，所述分离的抗原结合蛋白可以包含例如抗体来源的蛋白支架或具有移植的CDR或CDR衍生物的备选蛋白支架或人工支架。此类支架可以包括被引入例如以稳定抗原结合蛋白的三维结构的突变的抗体来源的支架以及包含例如生物相容性聚合物的完全合成的支架。参见例如Korndorfer等,2003,Proteins:Structure,Function,and Bioinformatics,53(1):121-129(2003)；Roque等,Biotechnol.Prog.20:639-654(2004)。此外，肽抗体模拟物(PAMs)以及利用纤连蛋白组分基于抗体模拟物的支架可以用作支架。在本申请中，所述分离的抗原结合蛋白可以以7x10 -12或更低的KD值结合源自人和猴的GITR蛋白，其中所述KD值可以通过BLI法测定。 In this application, the term "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. Examples of 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. In the present application, 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. For example, 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. See, eg, Korndorfer et al., 2003, Proteins: Structure, Function, and Bioinformatics, 53(1):121-129 (2003); Roque et al., Biotechnol. Prog. 20:639-654 (2004). In addition, peptide antibody mimetics (PAMs) as well as scaffolds based on antibody mimetics utilizing fibronectin components can be used as scaffolds. In the present application, 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.
在本申请中，术语“GITR”通常指“糖皮质激素诱导的TNF相关基因”，也称为TNF受体超家族18(TNFRSF18)、AITR或CD357。在本申请中，所述GITR可以与GITR配体(GITRL)结合。所述GITR可以包括由细胞天然表达的GITR的任何变体或同种型。例如，人源的GITR可以包括3种变体，其氨基酸序列可以分别如登录号NP_004186、NP_683699和NP_683700所示。人和鼠形式的GITR的氨基酸和核酸序列描述于WO98/06842中，其以引用的方式并入本申请。也参见GenBank登录号Q9Y5U5(人来源的氨基酸序列)和AF109216(鼠来源的核酸和氨基酸序列)。成熟的人GITR多肽可以包含SEQ ID NO:49所示的氨基酸序列。来自食蟹猴的示例性成熟GITR蛋白可以包含SEQ ID NO:51所示的氨基酸序列。此外，术语“GITR”也包括天然存在的等位基因。In this application, the term "GITR" generally refers to "glucocorticoid-induced TNF-related gene", also known as TNF receptor superfamily 18 (TNFRSF18), AITR or CD357. In the present application, the GITR may bind to a GITR ligand (GITRL). The GITR can include any variant or isoform of the GITR that is naturally expressed by the cell. For example, 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. 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. Furthermore, the term "GITR" also includes naturally occurring alleles.
在本申请中，术语“GITRL”通常指GITR配体或其功能活性部分，和/或可溶性GITR配体。GITRL还可以包括GITRL的天然存在的等位基因变体、与天然存在的GITR配体分子在氨基酸序列上不同的GITR配体变体、以及这样的变体的组合，其中这些变体保留特异性地结合GITR受体的能力。In this application, the term "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.
在本申请中，术语“免疫检查点抑制剂”通常指能够抑制免疫检查点生物学活性的任何试剂。在本申请中，所述免疫检查点(immune cheeckpoint)可以为在免疫***中起抑制作用的调节分子。所述免疫检查点可以表达于免疫细胞上，抑制免疫细胞的功能。所述免疫检查点可以与肿瘤免疫逃逸相关。在本申请中，所述免疫检查点可以包括PD-1。在本申请中，所述 抑制剂可以包括能够抑制配体和受体(例如PD-1和PD-L1)之间相互作用的任何试剂。在本申请中，所述试剂可以为化学试剂，也可以为蛋白质和/或多肽。例如，所述试剂可以包括抗体或其抗原结合片段。In this application, the term "immune checkpoint inhibitor" generally refers to any agent capable of inhibiting the biological activity of an immune checkpoint. In the present application, 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. In the present application, the immune checkpoint may include PD-1. In the present application, the inhibitor can include any agent capable of inhibiting the interaction between a ligand and a receptor (e.g., PD-1 and PD-L1). In the present application, the reagents can be chemical reagents or proteins and/or polypeptides. For example, the agent may comprise an antibody or antigen-binding fragment thereof.
在本申请中，术语“PD-1”通常指CD28家族的免疫抑制受体。PD-1可以表达于预先活化的T细胞，并与两个配体PD-L1和/或PD-L2结合。在本申请中，所述PD-1可以包括人PD-1(hPD-1)或其变体、同种型以及物种同源物，以及与hPD-1具有至少一个共同表位的类似物。hPD-1的GenBank登录号为U64863。In this application, 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. In the present application, 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.
在本申请中，术语“PD-L1”通常指PD-1的一种细胞表面糖蛋白配体之一。所述PD-L1可以与PD-1结合。所述PD-L1可以与PD-1结合时下调T细胞的活化和/或细胞因子的分泌。在本申请中，所述PD-L1可以包括人PD-L(hPD-L1)或其变体、同种型以及物种同源物，以及与hPD-L1具有至少一个共同表位的类似物。hPD-L1的GenBank登录号为Q9NZQ7。In this application, 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. In the present application, 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.
在本申请中，术语“药物产品”通常指用于预防、缓解或治疗人的疾病(例如癌症)，有目的地调节人的生理机能并规定有适应症或者功能主治、用法和用量的物质。在本申请中，所述药物产品可以包括生物制品。例如，所述药物产品可以包括抗原结合蛋白。又例如，所述药物产品可以包括化学药。在本申请中，所述药物产品可以为混合物，也可以为组合物。在本申请中，所述药物产品可以以整体包装的形式存在，也可以被包装为多个包装。In this application, 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. In the present application, the drug product may include a biological product. For example, the drug product can include an antigen binding protein. As another example, the pharmaceutical product may include a chemical. In this application, the drug product may be a mixture or a composition. In the present application, the pharmaceutical product may exist in the form of an integral package, or may be packaged into multiple packages.
在本申请中，术语“GITR阳性的癌症”通常指肿瘤细胞(例如肿瘤浸润Treg细胞)表达GITR的癌症。In this application, the term "GITR-positive cancer" generally refers to cancers in which tumor cells (eg, tumor-infiltrating Treg cells) express GITR.
在本申请中，术语“PD-1阳性的癌症”通常指肿瘤细胞表达PD-1的癌症。In this application, the term "PD-1 positive cancer" generally refers to cancers in which tumor cells express PD-1.
在本申请中，术语“实体瘤”通常指可以通过临床检查(例如X线照射、CT扫描、B超或触诊等)手段检测到的有形的肿瘤。所述肿瘤可以包括异常细胞生长形成的赘生物或实体病变。In this application, the term "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.
在本申请中，术语“乳腺癌”通常指发生在乳腺腺上皮组织的癌症。In this application, the term "breast cancer" generally refers to cancer that occurs in the epithelial tissue of the breast glands.
在本申请中，术语“直肠癌”通常指发生在从齿状线至直肠乙状结肠交界处之间的癌症。In this application, the term "rectal cancer" generally refers to cancer that occurs from the dentate line to the rectosigmoid junction.
在本申请中，术语“结肠癌”通常指发生于结肠部位的消化道的癌症。例如，所述结肠癌可以发生于直肠与乙状结肠交界处。In this application, the term "colon cancer" generally refers to cancer of the digestive tract that occurs in the colon. For example, the colon cancer can occur at the junction of the rectum and sigmoid colon.
在本申请中，术语“可变结构域”通常指抗体重链或轻链的氨基末端结构域。重链和轻链的可变结构域可以分别称为“VH”和“VL”。这些结构域通常是抗体的变化最大的部分(相对于相同类型的其它抗体)，且包含抗原结合位点。In this application, the term "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.
在本申请中，术语“可变”通常指在抗体之间可变结构域的某些区段在序列上存在很大差 异的事实。V结构域介导抗原结合并决定特定抗体对其特定抗原的特异性。然而，可变性并非在整个可变结构域范围内均匀分布。相反，它可以集中在轻链和重链可变结构域中称为高变区(CDR或HVR)的三个区段中。可变结构域的更高度保守的部分称为框架区(FR)。天然重链和轻链的可变结构域可以各自包含四个FR区，大多数采用β-折叠构型，通过三个CDR连接，其形成环形连接，并且在一些情况下形成β-折叠结构的一部分。每条链中的CDR可以通过FR区紧密靠近地保持在一起，并且来自另一条链的CDR一同促进抗体的抗原结合位点的形成(参见Kabat et al,Sequences of Immunological Interest,Fifth Edition,National Institute of Health,Bethesda,Md.(1991))。恒定结构域可以不直接参与抗体与抗原的结合，但显示出各种效应子功能，例如抗体参与抗体依赖性细胞毒性。In this application, the term "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. However, the variability is not evenly distributed across the variable domains. Instead, it can be concentrated in three segments called hypervariable regions (CDRs or HVRs) in the light and heavy chain variable domains. 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 β-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.
在本申请中，术语“抗体”通常指免疫球蛋白或其片段或其衍生物，涵盖包括抗原结合位点的任何多肽，无论其是在体外还是体内产生的。该术语可以包括多克隆的、单克隆的、单特异性的、多特异性的、非特异性的、人源化的、单链的、嵌合的、合成的、重组的、杂化的、突变的和移植的抗体。除非另外被术语“完整的”修饰，如在“完整的抗体”中，为了本申请的目的，术语“抗体”也包括抗体片段，比如Fab、F(ab')2、Fv、scFv、Fd、dAb和保持抗原结合功能(例如，特异性结合GITR)的其它抗体片段。通常，这样的片段可以包括抗原结合结构域。基本的4链抗体单元是由两个相同的轻(L)链和两个相同的重(H)链组成的异四聚体糖蛋白。IgM抗体由5个基本的异四聚体单元与另外一个称为J链的多肽组成，且含有10个抗原结合位点，而IgA抗体包括2-5个可以与J链相结合聚合形成多价组合的基本4链单元。就IgG而言，4链单元一般为约150,000道尔顿。每个L链通过一个共价二硫键与H链连接，而两个H链通过一个或多个取决于H链同种型的二硫键相互连接。每个H和L链还具有规则间隔的链内二硫化桥键。每个H链在N末端具有可变结构域(VH)，对于α和γ链各自继之以三个恒定结构域(CH)、对于μ和ε同种型继之以四个CH结构域。每个L链在N末端具有可变结构域(VL)，在其另一端具有恒定结构域。VL与VH对应，且CL与重链的第一恒定结构域(CH1)相对应。特定的氨基酸残基可以被认为在轻链和重链可变结构域之间形成界面。VH和VL可以配对一起形成单个抗原结合位点。对于不同类别抗体的结构和性质，参见例如Basic and Clinical Immunology,8th Edition,Daniel P.Sties,Abba I.Terr and Tristram G.Parsolw(eds),Appleton & Lange,Norwalk,Conn.,1994，第71页和第6章。来自任何脊椎动物物种的L链可以基于其恒定结构域的氨基酸序列被分为两种明显不同的类型中的一种，称为κ和λ。取决于其重链(CH)恒定结构域的氨基酸序列，可以将免疫球蛋白分为不同的类别或同种型。存在五类免疫球蛋白：IgA、IgD、IgE、IgG和IgM，具有分别被命名 为α、δ、ε、γ和μ的重链。基于CH序列和功能方面的相对小的差异，将γ和α类进一步分成亚类，例如，人可以表达下述亚类：IgG1、IgG2A、IgG2B、IgG3、IgG4、IgA1和IgK1。In this application, the term "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. Unless otherwise modified by the term "intact", as in "intact antibody", for the purposes of this application, 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. For IgG, 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. For the structure and properties of different classes of antibodies, see, eg, Basic and Clinical Immunology, 8th Edition, Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds), Appleton & Lange, Norwalk, Conn., 1994, p. 71 page and Chapter 6. L-chains 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.
在本申请中，术语“CDR”通常指抗体可变结构域的区域，其序列是高度可变的和/或形成结构定义环。通常，抗体可以包括六个CDR；在VH中三个(HCDR1、HCDR2、HCDR3)，和在VL中三个(LCDR1、LCDR2、LCDR3)。在天然抗体中，HCDR3和LCDR3可以显示所述六个CDR的大多数多样性，并且特别地HCDR3被认为在赋予抗体的精细特异性方面起独特作用。参见，例如Xu et al,Immunity 13:37-45(2000)；Johnson and Wu,in Methods in Molecular Biology 248:1-25(Lo,ed.,Human Press,Totowa,N.J.,2003)。实际上，仅由重链组成的天然存在的骆驼抗体在缺乏轻链的情况功能正常且稳定。例如,Hamers-Casterman et al.,Nature 363:446-448(1993)；Sheriff et al,Nature Struct.Biol.3:733-736(1996)。In this application, the term "CDR" generally refers to regions of antibody variable domains whose sequences are highly variable and/or form structurally defined loops. Typically, an antibody may include six CDRs; three in the VH (HCDRl, HCDR2, HCDR3), and three in the VL (LCDRl, LCDR2, LCDR3). Among native antibodies, 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. See, e.g., Xu et al, Immunity 13:37-45 (2000); Johnson and Wu, in Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, N.J., 2003). In fact, naturally occurring camelid antibodies consisting only of heavy chains function normally and are stable in the absence of light chains. For example, Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al, Nature Struct. Biol. 3:733-736 (1996).
在本申请中，术语“FR”通常指抗体可变结构域的更高度保守的部分，其被称为框架区。通常，天然重链和轻链的可变结构域可以各自包含四个FR区，即在VH中四个(H-FR1，H-FR2，H-FR3，和H-FR4)，和在VL中四个(L-FR1，L-FR2，L-FR3，和L-FR4)。例如，本申请所述的分离的抗原结合蛋白的VL可以包括框架区L-FR1，L-FR2，L-FR3，和L-FR4。本申请所述的分离的抗原结合蛋白的VH可以包括框架区H-FR1，H-FR2，H-FR3，和H-FR4。In this application, the term "FR" generally refers to the more highly conserved portions of antibody variable domains, which are referred to as framework regions. Typically, the 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). For example, the 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.
在本申请中，术语“抗原结合片段”通常指具有抗原结合活性的片段。在本申请中，所述抗原结合片段可以包括Fab，Fab’，F(ab)2、Fv片段、F(ab’)2，scFv，di-scFv和/或dAb。In the present application, the term "antigen-binding fragment" generally refers to a fragment having antigen-binding activity. In the present application, the antigen-binding fragment may include Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
在本申请中，术语“Fab”通常是指含有重链可变结构域和轻链可变结构域的片段，并且还含有轻链的恒定结构域和重链的第一恒定结构域(CH1)；术语“Fab’”通常是指在重链CH1结构域的羧基端添加少量残基(包括一个或多个来自抗体铰链区的半胱氨酸)而不同于Fab的片段；术语“F(ab') 2”通常是指Fab’的二聚体，包含通过铰链区上的二硫桥连接的两个Fab片段的抗体片段。术语“Fv”通常是指含有完整抗原识别与结合位点的最小抗体片段。在某些情形中，该片段可以由一个重链可变区和一个轻链可变区以紧密非共价结合的二聚体组成；术语“dsFv”通常是指二硫键稳定的Fv片段，其单个轻链可变区与单个重链可变区之间的键是二硫键。术语“dAb片段”通常是指由VH结构域组成的抗体片段。在本申请中，术语“scFv”通常是指抗体的一个重链可变结构域和一个轻链可变结构域通过柔性肽连接子共价连接配对形成的单价分子；此类scFv分子可具有一般结构：NH 2-VL-连接子-VH-COOH或NH 2-VH-连接子-VL-COOH。 In this application, the term "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 The term "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; the term "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. The term "Fv" generally refers to the smallest antibody fragment containing the entire antigen recognition and binding site. In certain instances, the fragment may consist of a heavy chain variable region and a light chain variable region in a tightly non-covalently bound dimer; the term "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. The term "dAb fragment" generally refers to antibody fragments consisting of VH domains. In this application, the term "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.
在本申请中，术语“人源化抗体”通常是指非人抗体(例如小鼠抗体)的CDR区以外的部分或全部有的氨基酸被源自人免疫球蛋白的相应的氨基酸置换的抗体。在CDR区中，氨基酸 的小的添加、缺失、***、置换或修饰也可以是允许的，只要它们仍保留抗体结合特定抗原的能力。人源化抗体可任选地包含人类免疫球蛋白恒定区的至少一部分。“人源化抗体”保留类似于原始抗体的抗原特异性。非人(例如鼠)抗体的“人源化”形式可以最低限度地包含衍生自非人免疫球蛋白的序列的嵌合抗体。在某些情形中，可以将人免疫球蛋白(受体抗体)中的CDR区残基用具有所期望性质、亲和力和/或能力的非人物种(供体抗体)(诸如小鼠，大鼠，家兔或非人灵长类动物)的CDR区残基替换。在某些情形中，可以将人免疫球蛋白的FR区残基用相应的非人残基替换。此外，人源化抗体可包含在受体抗体中或在供体抗体中没有的氨基酸修饰。进行这些修饰可以是为了进一步改进抗体的性能，诸如结合亲和力。In the present application, the term "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. In some cases, CDR region residues in a human immunoglobulin (acceptor antibody) 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. In certain instances, FR region residues of the human immunoglobulin can be replaced with corresponding non-human residues. In addition, 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.
在本申请中，术语“直接相连”可以与术语“间接相连”相对，术语“直接相连”通常是指直接连接。例如，所述直接相连可以为物质间没有间隔子而直接相连的情况。所述间隔子可以是连接子。例如，所述连接子可以为肽连接子。术语“间接相连”通常是指物质间不直接相连的情况。例如，所述间接相连可以为通过间隔子而连接的情况。例如，在本申请所述的分离的抗原结合蛋白中，所述L-FR1的C末端与所述LCDR1的N末端可以直接或间接相连。In this application, the term "directly connected" may be opposed to the term "indirectly connected", which generally refers to a direct connection. For example, the direct connection may be the case where substances are directly connected without a spacer. The spacer may be a linker. For example, the linker can be a peptide linker. The term "indirectly connected" generally refers to a situation where substances are not directly connected. For example, the indirect linkage may be the case of linkage through a spacer. For example, in the isolated antigen binding protein described in the present application, the C-terminus of L-FR1 and the N-terminus of LCDR1 may be linked directly or indirectly.
在本申请中，术语“分离的核酸分子”通常指任何长度的分离形式的核苷酸，脱氧核糖核苷酸或核糖核苷酸，或从其天然环境分离的或人工合成的类似物。In this application, the term "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.
在本申请中，术语“载体”通常指可将编码某蛋白的多聚核苷酸***其中并使蛋白获得表达的一种核酸运载工具。载体可通过转化、转导或转染宿主细胞，使其携带的遗传物质元件在宿主细胞内表达得以表达。例如，所述载体可以为质粒。一种载体可能含有多种控制表达的元件，包括启动子序列、转录起始序列、增强子序列、选择元件及报告基因。另外，载体还可含有复制起始位点。载体还有可能包括有协助其进入细胞的成分，如病毒颗粒、脂质体或蛋白外壳，但不仅仅只有这些物质。In this application, 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. For example, 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.
在本申请中，术语“细胞”通常指可以是或已经是受试者质粒或载体的接受者的单个细胞、细胞系或细胞培养物，其包括本申请所述的核酸分子或本申请所述的载体。细胞可以包括单个细胞的后代。由于天然、偶然或有意的突变，后代可以不一定与原始母细胞完全相同(在总DNA互补体的形态上或在基因组上)。细胞可包括用本申请所述的载体在体外转染的细胞。细胞可以是细菌细胞(例如，大肠杆菌)、酵母细胞或其它真核细胞，例如，所述细胞可以为哺乳动物细胞。In this application, 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.
在本申请中，术语“药物组合物”通常指涉及适合施用于患者、例如人患者的组合物。例如，本申请所述的药物组合物，其可以包含本申请所述的分离的抗原结合蛋白、本申请所述的核酸分子、本申请所述的载体和/或本申请所述的细胞，以及任选地药学上可接受的佐剂。 此外，所述药物组合物还可以包含一种或多种(药学上有效的)载剂、稳定剂、赋形剂、稀释剂、增溶剂、表面活性剂、乳化剂和/或防腐剂的合适的制剂。组合物的可接受成分在所用剂量和浓度下可以对接受者无毒。本申请的药物组合物包括但不限于液体、冷冻和冻干组合物。在本申请中，术语“药学上可接受的佐剂”通常指与药物给药相容的任何和所有的溶剂、分散介质、包衣、等渗剂和吸收延迟剂等，通常安全、无毒，且既不是生物学上也非其它方面不合需要的。In this application, the term "pharmaceutical composition" generally refers to a composition suitable for administration to a patient, eg, a human patient. For example, 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. In addition, 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. Pharmaceutical compositions of the present application include, but are not limited to, liquid, frozen, and lyophilized compositions. In this application, the term "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.
在本申请中，术语“受试者”通常指人类或非人类动物，包括但不限于猫、狗、马、猪、奶牛、羊、兔、小鼠、大鼠或猴。In this application, 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.
在本申请中，术语“包含”通常是指包括明确指定的特征，但不排除其他要素。In this application, the term "comprising" generally means including the expressly specified features, but not excluding other elements.
在本申请中，术语“约”通常是指在指定数值以上或以下0.5％-10％的范围内变动，例如在指定数值以上或以下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％、或10％的范围内变动。In this application, 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%.
发明详述Detailed description of the invention
靶向GITR的抗原结合蛋白Antigen-binding proteins targeting GITR
本申请所涉及的靶向GITR的抗原结合蛋白可以包含氨基酸序列如SEQ ID NO:23所示的VH中的至少一个CDR。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.
例如，本申请所述的靶向GITR的抗原结合蛋白，其可以包含氨基酸序列如SEQ ID NO:23所示的VH中的HCDR3。例如，本申请所述的靶向GITR的抗原结合蛋白，其可以包含氨基酸序列如SEQ ID NO:23所示的VH中的HCDR2。又例如，本申请所述的靶向GITR的抗原结合蛋白，其可以包含氨基酸序列如SEQ ID NO:23所示的VH中的HCDR1。For example, 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. For example, 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. For another example, 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.
需要说明的是，在本申请中，本申请所述的靶向GITR的抗原结合蛋白的VL和/或VH中的CDR可以按照Kabat进行定义。It should be noted that, in the present application, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白可以包含氨基酸序列如SEQ ID NO:24所示的VL中的至少一个CDR。例如，本申请所述的靶向GITR的抗原结合蛋白，其可以包含氨基酸序列如SEQ ID NO:24所示的VL中的LCDR1。例如，本申请所述的靶向GITR的抗原结合蛋白，其可以包含氨基酸序列如SEQ ID NO:24所示的VL中的LCDR2。又例如，本申请所述的靶向GITR的抗原结合蛋白，其可以包含氨基酸序列如SEQ ID NO:24所示的VL中的LCDR3。In the present application, 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. For example, 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. For example, 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. In another example, 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.
本申请所述的靶向GITR的抗原结合蛋白中，所述HCDR3可以包含SEQ ID NO:2所示的氨基酸序列。In the antigen binding protein targeting GITR described in this application, the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 2.
本申请所述的靶向GITR的抗原结合蛋白中，所述HCDR1可以包含SEQ ID NO:3所示的氨基酸序列。In the antigen binding protein targeting GITR described in this application, the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 3.
本申请所述的靶向GITR的抗原结合蛋白中，所述HCDR2可以包含SEQ ID NO:4所示的氨基酸序列。In the antigen binding protein targeting GITR described in this application, the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 4.
本申请所述的靶向GITR的抗原结合蛋白中，所述LCDR1可以包含SEQ ID NO:10所示的氨基酸序列。In the antigen binding protein targeting GITR described in this application, the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 10.
本申请所述的靶向GITR的抗原结合蛋白中，所述LCDR2可以包含SEQ ID NO:11所示的氨基酸序列。In the antigen binding protein targeting GITR described in this application, the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 11.
本申请所述的靶向GITR的抗原结合蛋白中，所述LCDR3可以包含SEQ ID NO:12所示的氨基酸序列。In the antigen binding protein targeting GITR described in this application, the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 12.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白，其可以包括抗体或其抗原结合片段。例如，本申请所述的靶向GITR的抗原结合蛋白可以包括但不限于重组抗体、单克隆抗体、人抗体、人源化抗体、嵌合抗体、双特异性抗体、单链抗体、双抗体、三抗体、四抗体、Fv片段、scFv片段、Fab片段、Fab'片段、F(ab')2片段和骆驼化单结构域抗体。In the present application, the antigen-binding proteins targeting GITR described in the present application may include antibodies or antigen-binding fragments thereof. For example, 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.
在本申请中，所述抗体可以为人源化抗体。换句话说，本申请所述的靶向GITR的抗原结合蛋白，其可以为免疫特异性结合至相关抗原(例如人类GITR))且包含基本上具有人类抗体的氨基酸序列的框架(FR)区及基本上具有非人类抗体的氨基酸序列的互补决定区(CDR)的抗体或其变异体、衍生物、类似物或片段。此处的“基本上”在CDR的情况下是指CDR的氨基酸序列与非人类抗体CDR的氨基酸序列至少80％、至少85％、至少90％、至少95％、至少98％或至少99％同一。所述人源化抗体基本上可以包含所有至少一个且通常两个可变域(Fab、Fab′、F(ab′)2、FabC、Fv)，其中所有或基本上所有CDR区对应于非人类免疫球蛋白(即抗体)的CDR区且所有或基本上所有框架区为具有人类免疫球蛋白共有序列的框架区。例如，人源化抗体还包含至少一部分免疫球蛋白恒定区(例如，Fc)，通常为人类免疫球蛋白的恒定区。在一些实施方式中，人源化抗体含有轻链以及重链的至少可变域。抗体还可包括重链的CH1、铰链、CH2、CH3及CH4区。在一些实施方式中，人源化抗体仅含人源化轻链。在一些实施方式中，人源化抗体仅含人源化重链。在一些实施方式中，人源化抗体仅含轻链和/或人源化重链的人源化可变域。In the present application, the antibody may be a humanized antibody. In other words, 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. Here "substantially" in the context of a CDR means that the amino acid sequence of the CDR is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% identical to the amino acid sequence of a CDR of a non-human antibody . 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 (ie, an antibody) are framework regions having human immunoglobulin consensus sequences. For example, a humanized antibody also comprises at least a portion of an immunoglobulin constant region (eg, Fc), typically a human immunoglobulin constant region. In some embodiments, 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. In some embodiments, the humanized antibody contains only humanized light chains. In some embodiments, the humanized antibody contains only humanized heavy chains. In some embodiments, the humanized antibody contains only the humanized variable domains of the light chain and/or the humanized heavy chain.
在本申请中，所述抗原结合片段可以包括Fab，Fab’，F(ab)2、Fv片段、F(ab’)2，scFv，di-scFv和/或dAb。In the present application, the antigen-binding fragment may include Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白可以与参比抗体竞争结合GITR蛋白，其中所述参比抗体可以包含氨基酸序列如SEQ ID NO:23所示的VH中的HCDR1-3以及 氨基酸序列如SEQ ID NO:24所示的VL中的LCDR1-3。In the present application, 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.
在本申请中，所述参比抗体的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且所述参比抗体的LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。In the present application, 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.
在本申请中，所述参比抗体的轻链可变区可以包含SEQ ID NO:23所示的氨基酸序列；且所述参比抗体的重链可变区可以包含SEQ ID NO:24所示的氨基酸序列。In the present application, 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.
在本申请中，所述参比抗体的轻链可以包含SEQ ID NO:50、46-48中任一项所示的氨基酸序列；且所述参比抗体的重链可以包含SEQ ID NO:49、43-45中任一项所示的氨基酸序列。In the present application, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白的所述VL可以包括框架区L-FR1，L-FR2，L-FR3，和L-FR4。In the present application, the VL of the GITR-targeting antigen binding protein described herein may include framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR1的C末端可以与所述LCDR1的N末端直接或间接相连，且所述L-FR1可以包含SEQ ID NO:39所示的氨基酸序列。For example, 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.
DIVMTQSPLSLPVX 1LGX 2X 3ASISC(SEQ ID NO:39)，其中，X 1可以是S或T，X 2可以是Q或D，X 3可以是P或Q。 DIVMTQSPLSLPVX 1 LGX 2 X 3 ASISC (SEQ ID NO: 39), wherein X 1 can be S or T, X 2 can be Q or D, and X 3 can be P or Q.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR1可以包含SEQ ID NO:13、30、34中任一项所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR2可以位于所述LCDR1与所述LCDR2之间，且所述L-FR2可以包含SEQ ID NO:40所示的氨基酸序列。For example, 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 1PGQSPKLLIY(SEQ ID NO:40)，其中，X 1可以是R或K。 WYLQX 1 PGQSPKLLIY (SEQ ID NO: 40), wherein X 1 can be R or K.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR2可以包含SEQ ID NO:14、31中任一项所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR3可以位于所述LCDR2与所述LCDR3之间，且所述L-FR3可以包含SEQ ID NO:41所示的氨基酸序列。For example, 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 1GVYYC(SEQ ID NO:41)，其中，X 1可以为L或V。 GVPDRFSGSGSGTDFTLKISRVEAEDX 1 GVYYC (SEQ ID NO: 41), wherein X 1 can be L or V.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR3可以包含SEQ ID NO:15、32中任一项所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR4的N末端可以与所述LCDR3的C末端相连，且所述L-FR4可以包含SEQ ID NO:42所示的氨基酸序列。For example, 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 1EIK(SEQ ID NO:42)，其中，X 1可以是V或L。 FGGGTKX 1 EIK (SEQ ID NO: 42), wherein X 1 can be V or L.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述L-FR4可以包含SEQ ID NO:16、33中任一项所示的氨基酸序列。For example, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白的所述VL可以包含SEQ ID NO:24所示的氨基酸序列。In the present application, 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 1LGX 2X 3ASISCRSSQTIVHSNGNTYLEWYLQX 4PGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDX 5GVYYCFQGSHVPWTFGGGTKX 6EIK(SEQ ID NO:24)，其中，X 1可以是S或T，X 2可以是Q或D，X 3可以是P或Q，X 4可以是R或K，X 5可以为L或V，X 6可以是V或L。 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述VL可以包含SEQ ID NO:9、20-22中任一项所示的氨基酸序列。For example, the 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白可以包括抗体轻链恒定区，且所述抗体轻链恒定区可以包括人Igκ恒定区。In the present application, 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 Igκ constant region.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白的所述抗体轻链恒定区可以包含SEQ ID NO:52所示的氨基酸序列。In the present application, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白可以包含抗体轻链LC，且所述LC可以包含SEQ ID NO:50、46-48中任一项所示的氨基酸序列。In the present application, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白的所述VH可以包括框架区H-FR1，H-FR2，H-FR3，和H-FR4。In the present application, the VH of the GITR-targeting antigen binding protein described herein may include framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR1的C末端与所述HCDR1的N末端直接或间接相连，且所述H-FR1可以包含SEQ ID NO:35所示的氨基酸序列。For example, 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.
QVX 1LQESGPX 2X 3X 4X 5PX 6QTLX 7LTCX 8FSGFSLS(SEQ ID NO:35)，其中，X 1可以为T或Q，X 2可以为G或T，X 3可以为I或L，X 4可以为L或V，X 5可以为Q或K，X 6可以为S或T，X 7可以为S或T，X 8可以为S或T。 QVX 1 LQESGPX 2 X 3 X 4 X 5 PX 6 QTLX 7 LTCX 8 FSGFSLS (SEQ ID NO: 35), wherein X 1 can be T or Q, X 2 can be G or T, and 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR1可以包含SEQ ID NO:5、25、29中任一项所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR2可以位于所述HCDR1与所述HCDR2之间，且所述H-FR2可以包含SEQ ID NO:36所示的氨基酸序列。For example, 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 1GKGLEWLVLI(SEQ ID NO:36)，其中，X 1可以为P或S。 WIRQPX 1 GKGLEWLVLI (SEQ ID NO: 36), wherein X 1 can be P or S.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR2可以包含SEQ ID NO:6、26中任一项所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR3可以位于所述HCDR2与 所述HCDR3之间，且所述H-FR3可以包含SEQ ID NO:37所示的氨基酸序列。For example, 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 1KDTSX 2NQVX 3LX 4IX 5X 6X 7DX 8X 9DTATYYCAR(SEQ ID NO:37)，其中，X 1可以为S或T，X 2可以为N或K，X 3可以为F或V，X 4可以为K或T，X 5可以为A或T，X 6可以为S或N，X 7可以为V或M，X 8可以为T或P，X 9可以为A或V。 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 .
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR3可以包含SEQ ID NO:7、27中任一项所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR4的N末端可以与所述HCDR3的C末端相连，且所述H-FR4可以包含SEQ ID NO:38所示的氨基酸序列。For example, 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 1GTX 2VTVSS(SEQ ID NO:38)，其中，X 1可以是Q或T，X 2可以是M或T。 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述H-FR4可以包含SEQ ID NO:8、28中任一项所示的氨基酸序列。For example, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白的所述VH可以包含SEQ ID NO:23所示的氨基酸序列。In the present application, 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.
QVX 1LQESGPX 2X 3X 4X 5PX 6QTLX 7LTCX 8FSGFSLSTFGMGVGWIRQPX 9GKGLEWLVLILWNDIKYYNPALKSLLTVX 10KDTSX 11NQVX 12LX 13IX 14X 15X 16DX 17X 18DTATYYCARVDGYYGYFDVWGX 19GTX 20VTVSS(SEQ ID NO:23)，其中，X 1可以为T或Q，X 2可以为G或T，X 3可以为I或L，X 4可以为L或V，X 5可以为Q或K，X 6可以为S或T，X 7可以为S或T，X 8可以为S或T，X 9可以为P或S，X 10可以为S或T，X 11可以为N或K，X 12可以为F或V，X 13可以为K或T，X 14可以为A或T，X 15可以为S或N，X 16可以为V或M，X 17可以为T或P，X 18可以为A或V，X 19可以是Q或T，X 20可以是M或T。 QVX 1 LQESGPX 2 X 3 X 4 X 5 PX 6 QTLX 7 LTCX 8 FSGFSLSTFGMGVGWIRQPX 9 GKGLEWLVLILWNDIKYYNPALKSLLTVX 10 KDTSX 11 NQVX 12 LX 13 IX 14 X 15 X 16 DX 17 X 18 DTATYYCARVDGYYGYFDVWGX 19 ) , 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, X 17 can be T or P, X 18 can be A or V, X 19 can be Q or T, X 20 can be M or T.
例如，本申请所述的靶向GITR的抗原结合蛋白的所述VH可以包含SEQ ID NO:1、17-19中任一项所示的氨基酸序列。For example, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白可以包括抗体重链恒定区，且所述抗体重链恒定区可以源自人IgG重链恒定区。在其他一些实施方式中，本申请所述的靶向GITR的抗原结合蛋白可以包括抗体重链恒定区，且所述抗体重链恒定区可以源自人IgG1重链恒定区。In the present application, 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. In some other embodiments, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白的所述抗体重链恒定区可以包含SEQ ID NO:51所示的氨基酸序列。In the present application, 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.
在本申请中，本申请所述的靶向GITR的抗原结合蛋白可以包含抗体重链HC，且所述HC可以包含SEQ ID NO:49、43-45中任一项所示的氨基酸序列。In the present application, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:13所示的 氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:15所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:5所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:7所示的氨基酸序列。L-FR4可包含SEQ ID NO:8所示的氨基酸序列。For example, 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, and 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:31所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:33所示的氨基酸序列，且H-FR1可包含SEQ ID NO:25所示的氨基酸序列。H-FR2可包含SEQ ID NO:26所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:25所示的氨基酸序列。H-FR2可包含SEQ ID NO:26所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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, and 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:34所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:25所示的氨基酸序列。H-FR2可包含SEQ ID NO:26所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:31所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:33所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:34所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32 所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:31所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:33所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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.
例如，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:34所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。For example, 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, 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.
本申请所述的靶向GITR的抗原结合蛋白可包含抗体轻链可变区VL和抗体重链可变区VH。例如，所述VL可包含SEQ ID NO:9所示的氨基酸序列，所述VH可包含SEQ ID NO:1所示的氨基酸序列。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. For example, the VL can comprise the amino acid sequence set forth in SEQ ID NO:9, and the VH can comprise the amino acid sequence set forth in SEQ ID NO:1.
例如，所述VL可包含SEQ ID NO:20所示的氨基酸序列，所述VH可包含SEQ ID NO:17所示的氨基酸序列。For example, 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.
例如，所述VL可包含SEQ ID NO:21所示的氨基酸序列，所述VH可包含SEQ ID NO:17所示的氨基酸序列。For example, 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.
例如，所述VL可包含SEQ ID NO:22所示的氨基酸序列，所述VH可包含SEQ ID NO:17所示的氨基酸序列。For example, 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.
例如，所述VL可包含SEQ ID NO:20所示的氨基酸序列，所述VH可包含SEQ ID NO:18所示的氨基酸序列。For example, 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:18.
例如，所述VL可包含SEQ ID NO:21所示的氨基酸序列，所述VH可包含SEQ ID NO:18所示的氨基酸序列。For example, 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.
例如，所述VL可包含SEQ ID NO:22所示的氨基酸序列，所述VH可包含SEQ ID NO:18所示的氨基酸序列。For example, 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.
例如，所述VL可包含SEQ ID NO:20所示的氨基酸序列，所述VH可包含SEQ ID NO:19所示的氨基酸序列。For example, 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.
例如，所述VL可包含SEQ ID NO:21所示的氨基酸序列，所述VH可包含SEQ ID NO:19所示的氨基酸序列。For example, 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.
例如，所述VL可包含SEQ ID NO:22所示的氨基酸序列，所述VH可包含SEQ ID NO:19所示的氨基酸序列。For example, 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.
本申请所述的靶向GITR的抗原结合蛋白可包含抗体轻链和抗体重链。The GITR-targeting antigen binding proteins described herein may comprise an antibody light chain and an antibody heavy chain.
例如，所述轻链可包含SEQ ID NO:50所示的氨基酸序列，所述重链可包含SEQ ID NO:49所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:46所示的氨基酸序列，所述重链可包含SEQ ID NO:43所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:47所示的氨基酸序列，所述重链可包含SEQ ID NO:43所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:48所示的氨基酸序列，所述重链可包含SEQ ID NO:43所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:46所示的氨基酸序列，所述重链可包含SEQ ID NO:44所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:47所示的氨基酸序列，所述重链可包含SEQ ID NO:44所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:48所示的氨基酸序列，所述重链可包含SEQ ID NO:44所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:46所示的氨基酸序列，所述重链可包含SEQ ID NO:45所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:47所示的氨基酸序列，所述重链可包含SEQ ID NO:45所示的氨基酸序列。For example, 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.
例如，所述轻链可包含SEQ ID NO:48所示的氨基酸序列，所述重链可包含SEQ ID NO:45所示的氨基酸序列。For example, 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.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO: 10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:13所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:15所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:5所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:7所示的氨基酸序列。L-FR4可包含SEQ ID NO:8所示的氨基酸序列。所述VL可包含SEQ ID NO:9所示的氨基酸序列，所述VH可包含SEQ ID NO:1所示的氨基酸序列。所述轻链可包含SEQ ID NO:50所示的氨基酸序列，所述重链可包含SEQ ID NO:49所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为C3E2。In the present application, 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. Wherein, 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, and 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, and 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. For example, the antigen binding protein targeting GITR can be C3E2.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:31所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:33所示的氨基酸序列，且H-FR1可包含SEQ ID NO:25所示的氨基酸序列。H-FR2可包含SEQ ID NO:26所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:20所示的氨基酸序列，所述VH可包含SEQ ID NO:17所示的氨基酸序列。所述轻链可包含SEQ ID NO:46所示的氨基酸序列，所述重链可包含SEQ ID NO:43所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 1-2。In the present application, 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. Wherein, 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, 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. 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. For example, the antigen binding protein targeting GITR can be 3E2 1-2.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:25所示的氨基酸序列。H-FR2可包含SEQ ID NO:26所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:21所示的氨基酸序列，所述VH可包含SEQ ID NO:17所示的氨基酸序列。所述轻链可包含SEQ ID NO:47所示的氨基酸序列，所述重链可包含SEQ ID NO:43所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 1-3。In the present application, 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. Wherein, 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, and 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. For example, the antigen binding protein targeting GITR can be 3E2 1-3.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:34所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:25所示的氨基酸序列。H-FR2可包含SEQ ID NO:26所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:22所示的氨基酸序列，所述VH可包含SEQ ID NO:17所示的氨基酸序列。所述轻链可包含SEQ ID NO:48所示的氨基酸序列，所述重链可包含SEQ ID NO:43所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 1-4。In the present application, 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. Wherein, 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, and 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. For example, the antigen binding protein targeting GITR can be 3E2 1-4.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:31所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:33所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:20所示的氨基酸序列，所述VH可包含SEQ ID NO:18所示的氨基酸序列。所述轻链可包含SEQ ID NO:46所示的氨基酸序列，所述重链可包含SEQ ID NO:44所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 2-2。In the present application, 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. Wherein, 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, 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. 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. For example, the antigen binding protein targeting GITR can be 3E2 2-2.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:21所示的氨基酸序列，所述VH可包含SEQ ID NO:18所示的氨基酸序列。所述轻链可包含SEQ ID NO:47 所示的氨基酸序列，所述重链可包含SEQ ID NO:44所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 2-3。In the present application, 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. Wherein, 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, 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. 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. For example, the antigen binding protein targeting GITR can be 3E2 2-3.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:34所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:22所示的氨基酸序列，所述VH可包含SEQ ID NO:18所示的氨基酸序列。所述轻链可包含SEQ ID NO:48所示的氨基酸序列，所述重链可包含SEQ ID NO:44所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 2-4。In the present application, 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. Wherein, 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, 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. 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. For example, the antigen binding protein targeting GITR can be 3E2 2-4.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:31所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:33所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:20所示的氨基酸序列，所述VH可包含SEQ ID NO:19所示的氨基酸序列。所述轻链可包含SEQ ID NO:46所示的氨基酸序列，所述重链可包含SEQ ID NO:45所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 3-2。In the present application, 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. Wherein, 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, 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. 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. For example, the antigen binding protein targeting GITR can be 3E2 3-2.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:30所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L- FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:21所示的氨基酸序列，所述VH可包含SEQ ID NO:19所示的氨基酸序列。所述轻链可包含SEQ ID NO:47所示的氨基酸序列，所述重链可包含SEQ ID NO:45所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 3-3。In the present application, 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. Wherein, 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, 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. 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. For example, the antigen binding protein targeting GITR can be 3E2 3-3.
在本申请中，所述的靶向GITR的抗原结合蛋白的HCDR1-3可以分别包含SEQ ID NO:2、SEQ ID NO:3和SEQ ID NO:4所示的氨基酸序列，且LCDR1-3可以分别包含SEQ ID NO:10、SEQ ID NO:11和SEQ ID NO:12所示的氨基酸序列。其中，本申请所述的靶向GITR的抗原结合蛋白的L-FR1可包含SEQ ID NO:34所示的氨基酸序列，L-FR2可包含SEQ ID NO:14所示的氨基酸序列，L-FR3可包含SEQ ID NO:32所示的氨基酸序列，L-FR4可包含SEQ ID NO:16所示的氨基酸序列，且H-FR1可包含SEQ ID NO:29所示的氨基酸序列。H-FR2可包含SEQ ID NO:6所示的氨基酸序列。L-FR3可包含SEQ ID NO:27所示的氨基酸序列。L-FR4可包含SEQ ID NO:28所示的氨基酸序列。所述VL可包含SEQ ID NO:22所示的氨基酸序列，所述VH可包含SEQ ID NO:19所示的氨基酸序列。所述轻链可包含SEQ ID NO:48所示的氨基酸序列，所述重链可包含SEQ ID NO:45所示的氨基酸序列。例如，所述靶向GITR的抗原结合蛋白可以为3E2 3-4。In the present application, 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. Wherein, 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, 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. 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. For example, the antigen binding protein targeting GITR can be 3E2 3-4.
在本申请中，所述的靶向GITR的抗原结合蛋白可以具有下述性质中的一种或多种：1)能够以7x10 -12或更低的K D值结合源自人和猴的GITR蛋白，其中所述K D值通过BLI法测定；2)能够刺激免疫细胞增殖；3)能够刺激免疫细胞分泌IFN-γ，所述分泌在T细胞活性测定中测得；4)能够抑制肿瘤生长和/或肿瘤细胞增殖；5)能够激活GITR信号通路；6)能够抑制GITR与GITRL结合。 In the present application, 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-γ, 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.
在本申请中，所述的靶向GITR的抗原结合蛋白能够以7x10 -12M或更低的K D值结合源自人和猴的GITR蛋白，其中所述K D值可以通过BLI法测定。例如，本申请所述的靶向GITR的抗原结合蛋白结合源自人的GITR蛋白的K D值可以为≤7x10 -12M、≤6x10 -12M、≤5x10 -12M、≤4x10 -12M、≤3x10 -12M、≤2x10 -12M、≤1x10 -12M、≤0.5x10 -12M、≤0.1x10 -12M、≤0.01x10 -12M、≤0.05x10 -12M，或≤0.001x10 -12M。又例如，本申请所述的靶向GITR的抗原结合蛋白结合源自猴的GITR蛋白的K D值可以为≤7x10 -12M、≤6x10 -12M、≤5x10 -12M、≤4x10 -12M、≤3x10 -12M、≤2x10 -12M、≤1x10 -12M、≤0.5x10 -12M、≤0.1x10 -12M、≤0.01x10 -12M、≤0.05x10 -12M，或≤0.001x10 -12M。此外，本申请所述的靶向GITR的抗原结合蛋白结合源自鼠的GITR蛋白的K D值可以为≤7x10 -12M、≤6x10 -12M、≤5x10 -12M、≤4x10 -12M、≤3x10 -12M、≤2x10 -12M、≤1x10 - 12M、≤0.5x10 -12M、≤0.1x10 -12M、≤0.01x10 -12M、≤0.05x10 -12M，或≤0.001x10 -12M。在本申请中，所述K D值还可以通过ELISA、竞争ELISA或BIACORE或KINEXA进行测定。 In the present application, 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. For example, 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 . For another example, 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 . In addition, 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 . In the present application, the K D value can also be determined by ELISA, competitive ELISA or BIACORE or KINEXA.
在本申请中，所述的靶向GITR的抗原结合蛋白能够刺激免疫细胞增殖。例如本申请所述的靶向GITR的抗原结合蛋白能在体内或体外维持、提高、加速或延长免疫细胞增殖、生长和/或存活。可采用能检测细胞增殖、生长和/或存活的任何方法，例如细胞增殖试验或上皮屏障完整性试验(epithelial barrier integrity assay)来测定本申请所述的靶向GITR的抗原结合蛋白能否刺激免疫细胞增殖。在本申请中，所述免疫细胞可以选自下组一种或多种：淋巴细胞、天然杀伤细胞和髓样细胞。所述淋巴细胞可以为B细胞和/或T细胞。所述髓样细胞可以选自下组一种或多种：单核细胞、巨噬细胞、嗜曙红细胞、肥大细胞、嗜碱细胞和粒细胞。In the present application, the antigen binding protein targeting GITR can stimulate the proliferation of immune cells. For example, 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. In the present application, 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.
在本申请中，所述的靶向GITR的抗原结合蛋白能够刺激免疫细胞分泌IFN-γ，所述分泌可以在T细胞活性测定中测得，例如，可以采用ELISA、CBA或MSD法测定免疫细胞分泌的IFN-γ的含量。In the present application, the antigen binding protein targeting GITR can stimulate immune cells to secrete IFN-γ, 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-γ.
在本申请中，所述的靶向GITR的抗原结合蛋白能够预防、缓解或治疗癌症，例如，能够使GITR阳性的癌症的肿瘤体积减小至少约10％，至少约20％、至少约30％、至少约40％、至少约50％、至少约60％、至少约70％、至少约80％、至少约90％、至少约99％或100％。又例如，能够使肿瘤细胞数量减少至少约10％，至少约20％、至少约30％、至少约40％、至少约50％、至少约60％、至少约70％、至少约80％、至少约90％、至少约99％或100％。In the present application, 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%. In another example, 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%.
在本申请中，所述癌症可以包括GITR阳性的癌症；和/或，所述癌症包括PD-1阳性的癌症。In the present application, the cancer may comprise a GITR positive cancer; and/or, the cancer comprises a PD-1 positive cancer.
在本申请中，所述癌症可以包括实体瘤。In the present application, the cancer may include solid tumors.
在本申请中，述癌症可以选自下组：结肠癌、直肠癌和乳腺癌。In the present application, the cancer may be selected from the group consisting of colon cancer, rectal cancer and breast cancer.
在本申请中，所述的靶向GITR的抗原结合蛋白能够激活GITR信号通路，使得本申请所述的靶向GITR的抗原结合蛋白能够阻断GITRL及其受体GITR之间的相互作用，将T细胞的功能响应从功能失调状态修复为抗原刺激状态。In this application, 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.
在本申请中，所述的靶向GITR的抗原结合蛋白能够抑制GITR与GITRL结合，阻断GITRL及其受体GITR之间的相互作用，将T细胞的功能响应从功能失调状态修复为抗原刺激状态。In the present application, 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.
免疫检查点抑制剂immune checkpoint inhibitors
在本申请中，所述免疫检查点抑制剂可以阻碍PD-1和PD-L1的相互作用。In the present application, the immune checkpoint inhibitor can block the interaction of PD-1 and PD-L1.
在本申请中，所述免疫检查点抑制剂可以包含PD-1抑制剂。In the present application, the immune checkpoint inhibitor may comprise a PD-1 inhibitor.
在本申请中，所述免疫检查点抑制剂可以包含PD-1抗体或其抗原结合片段。In the present application, the immune checkpoint inhibitor may comprise a PD-1 antibody or an antigen-binding fragment thereof.
例如，所述免疫检查点抑制剂可以包含小鼠PD-1抗体或其抗原结合片段。又例如，所述 免疫检查点抑制剂可以包含人PD-1抗体或其抗原结合片段。例如，所述PD-1抗体或其抗原结合片段可以选择下组：派姆单抗(Pembrolizumab，通用名Keytruda，K药，由默沙东制备)、纳武利尤单抗(Opdivo，通用名Nivolumab，O药，由百时美施贵宝制备)、特瑞普利单抗(由君实生物制备)、信迪利单抗(Sintilimab，由信达生物制备)、卡瑞利珠单抗(Camrelizumab，由恒瑞医药制备)和普雷利珠单抗(由百济神州制备)。For example, the immune checkpoint inhibitor can comprise a mouse PD-1 antibody or an antigen-binding fragment thereof. In another example, the immune checkpoint inhibitor can comprise a human PD-1 antibody or an antigen-binding fragment thereof. For example, 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).
其中，所述人PD-1抗体或其抗原结合片段可以包含VL，所述VL可以分别包含以下的氨基酸序列：Wherein, the human PD-1 antibody or its antigen-binding fragment may comprise VL, and the VL may comprise the following amino acid sequences respectively:
EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKR(SEQ ID NO:61)；或，EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKR (SEQ ID NO: 61); or,
Figure PCTCN2021142299-appb-000001
Figure PCTCN2021142299-appb-000001
其中，所述人PD-1抗体或其抗原结合片段可以包含VH，所述VH可以分别包含以下的氨基酸序列：Wherein, the human PD-1 antibody or its antigen-binding fragment may comprise VH, and the VH may comprise the following amino acid sequences respectively:
QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSS(SEQ ID NO:63)；或,QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSS(SEQ ID NO:63); or,
Figure PCTCN2021142299-appb-000002
Figure PCTCN2021142299-appb-000002
在本申请中，所述PD-1抗体或抗原结合片段包含HCDR3，其中所述PD-1抗体或抗原结合片段的HCDR3可以包含RMP1-14的HCDR3。其中，RMP1-14购自Bioxcell，货号为BE0146。In the present application, 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. Among them, RMP1-14 was purchased from Bioxcell, the product number is BE0146.
在本申请中，所述PD-1抗体或抗原结合片段包含HCDR2，其中所述PD-1抗体或抗原结合片段的HCDR2可以包含RMP1-14的HCDR2。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含HCDR1，其中所述PD-1抗体或抗原结合片段的HCDR1可以包含RMP1-14的HCDR1。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含LCDR3，其中所述PD-1抗体或抗原结合片段的LCDR3可以包含RMP1-14的LCDR3。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含LCDR2，其中所述PD-1抗体或抗原结合片段的LCDR2可以包含RMP1-14的LCDR2。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含LCDR1，其中所述PD-1抗体或抗原结合片段的所述LCDR1可以包含RMP1-14的LCDR1。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含VH，所述VH可以包含RMP1-14的VH。In the present application, the PD-1 antibody or antigen-binding fragment comprises VH, which may comprise the VH of RMP1-14.
在本申请中，所述PD-1抗体或抗原结合片段包含VL，所述VL可以包含RMP1-14的VL。In the present application, the PD-1 antibody or antigen-binding fragment comprises VL, which may comprise the VL of RMP1-14.
在本申请中，所述PD-1抗体或抗原结合片段包含HCDR3，其中所述PD-1抗体或抗原结合片段的HCDR3可以包含派姆单抗的HCDR3；或者，可以包含纳武利尤单抗的HCDR3。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含HCDR2，其中所述PD-1抗体或抗原结合片段的HCDR2可以包含派姆单抗的HCDR2；或者，可以包含纳武利尤单抗的HCDR2。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含HCDR1，其中所述PD-1抗体或抗原结合片段的HCDR1可以包含派姆单抗的HCDR1；或者，可以包含纳武利尤单抗的HCDR1。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含LCDR3，其中所述PD-1抗体或抗原结合片段的LCDR3可以包含派姆单抗的LCDR3；或者，可以包含纳武利尤单抗的LCDR3。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含LCDR2，其中所述PD-1抗体或抗原结合片段的LCDR2可以包含派姆单抗的LCDR2；或者，可以包含纳武利尤单抗的LCDR2。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含LCDR1，其中所述PD-1抗体或抗原结合片段的所述LCDR1可以包含派姆单抗的LCDR1；或者，可以包含纳武利尤单抗的LCDR1。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含VH，所述VH可以包含派姆单抗的VH；或者，可以包含纳武利尤单抗的VH。In the present application, 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.
在本申请中，所述PD-1抗体或抗原结合片段包含VL，所述VL可以包含派姆单抗的VL；或者，可以包含纳武利尤单抗的VL。In the present application, 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.
药物产品及其用途Pharmaceutical products and their uses
一方面，本申请提供一种药物产品，其包含本申请所述的靶向GITR的抗原结合蛋白和本申请所述的免疫检查点抑制剂。In one aspect, the present application provides a pharmaceutical product comprising the GITR-targeting antigen binding protein described herein and the immune checkpoint inhibitor described herein.
在本申请中，所述药物产品可以包括药物组合物。例如，所述药物组合物还可以包括药学上可接受的试剂。在本申请中，所述药学上可接受的试剂可以包括稳定剂、pH调节剂和/或赋形剂。例如，所述药物组合物可以具备剂型。在本申请中，所述剂型可以包括液体制剂。In the present application, the pharmaceutical product may comprise a pharmaceutical composition. For example, the pharmaceutical composition may also include a pharmaceutically acceptable agent. In the present application, the pharmaceutically acceptable agent may include stabilizers, pH adjusters and/or excipients. For example, the pharmaceutical composition may have a dosage form. In the present application, the dosage form may comprise a liquid formulation.
在本申请中，所述药物产品中所述靶向GITR的抗原结合蛋白和所述免疫检查点抑制剂 可以相互不混合。例如，所述药物产品中所述靶向GITR的抗原结合蛋白和所述免疫检查点抑制剂可以分别存在于不同的容器中。在本申请中，所述容器可以包括注射用玻璃瓶。In the present application, the GITR-targeting antigen binding protein and the immune checkpoint inhibitor in the drug product may not be mixed with each other. For example, the GITR-targeting antigen binding protein and the immune checkpoint inhibitor in the drug product may be present in separate containers. In the present application, the container may comprise a glass vial for injection.
一方面，本申请提供了本申请所述的药物产品在制备预防、缓解或治疗癌症的药物中的应用。In one aspect, 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.
一方面，本申请提供了本申请所述的靶向GITR的抗原结合蛋白和本申请所述的免疫检查点抑制剂在制备预防、缓解或治疗癌症的药物中的用途。In one aspect, 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.
一方面，本申请提供一种预防、缓解或治疗癌症的方法，其包括以下步骤：向有需要的受试者施用本申请所述的靶向GITR的抗原结合蛋白，和/或，向有需要的受试者施用本申请所述的免疫检查点抑制剂。In one aspect, 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.
在本申请中，所述方法可以包括以下步骤：向有需要的受试者施用本申请所述的药物产品。In the present application, the method may comprise the step of administering the pharmaceutical product described herein to a subject in need thereof.
一方面，本申请提供了本申请所述的靶向GITR的抗原结合蛋白，和/或，本申请所述的药物产品，其用于预防、缓解或治疗癌症。In one aspect, 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.
在本申请中，所述癌症可以包括GITR阳性的癌症；和/或，所述癌症可以包括PD-1阳性的癌症。In the present application, the cancer may comprise a GITR positive cancer; and/or the cancer may comprise a PD-1 positive cancer.
在本申请中，所述癌症可以包括实体瘤。In the present application, the cancer may include solid tumors.
在本申请中，所述癌症可以选自下组：结肠癌、直肠癌和乳腺癌。In the present application, the cancer may be selected from the group consisting of colon cancer, rectal cancer and breast cancer.
在本申请中，所述靶向GITR的抗原结合蛋白的给药剂量可以为约1mg/Kg-约10mg/Kg，例如，可以为约1mg/Kg-约9mg/Kg，约1mg/Kg-约8mg/Kg，约1mg/Kg-约7mg/Kg，约1mg/Kg-约6mg/Kg，约1mg/Kg-约5mg/Kg，约1mg/Kg-约3mg/Kg，约3mg/Kg-约6mg/Kg或约1mg/Kg-约3mg/Kg；例如，可以为约1mg/Kg、约2mg/Kg、约3mg/Kg、约4mg/Kg、约5mg/Kg、约6mg/Kg、约7mg/Kg、约8mg/Kg、约9mg/Kg或约10mg/Kg。In the present application, 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.
在本申请中，所述免疫检查点抑制剂的给药剂量可以为约1mg/Kg-约10mg/Kg，例如，可以为约1mg/Kg-约9mg/Kg，约1mg/Kg-约8mg/Kg，约1mg/Kg-约7mg/Kg，约1mg/Kg-约6mg/Kg，约1mg/Kg-约5mg/Kg，约1mg/Kg-约3mg/Kg，约3mg/Kg-约6mg/Kg或约1mg/Kg-约3mg/Kg；例如，可以为约1mg/Kg、约2mg/Kg、约3mg/Kg、约4mg/Kg、约5mg/Kg、约6mg/Kg、约7mg/Kg、约8mg/Kg、约9mg/Kg或约10mg/Kg。In the present application, 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.
在本申请中，所述靶向GITR的抗原结合蛋白的给药频率可以为每周2次。In the present application, the administration frequency of the antigen binding protein targeting GITR may be twice a week.
在本申请中，所述免疫检查点抑制剂的给药频率可以为每周2次。In the present application, the administration frequency of the immune checkpoint inhibitor may be twice a week.
在本申请中，所述靶向GITR的抗原结合蛋白的给药方式可以为注射。例如，可以为腹 腔注射，肌肉注射或静脉注射。In the present application, the administration mode of the antigen binding protein targeting GITR can be injection. For example, it can be intraperitoneal, intramuscular or intravenous.
在本申请中，所述免疫检查点抑制剂的给药方式可以为注射。例如，可以为腹腔注射，肌肉注射或静脉注射。In the present application, the administration mode of the immune checkpoint inhibitor can be injection. For example, it can be intraperitoneal, intramuscular or intravenous.
在本申请中，所述靶向GITR的抗原结合蛋白的浓度可以为约1mg/mL-约10mg/mL，例如，可以为约1mg/mL-约9mg/mL，约1mg/mL-约8mg/mL，约1mg/mL-约7mg/mL，约1mg/mL-约6mg/mL，约1mg/mL-约5mg/mL，约1mg/mL-约4mg/mL，约1mg/mL-约3mg/mL或约1mg/mL-约2mg/mL；例如可以为约1mg/mL、约1.5mg/mL、约2mg/mL、约2.5mg/mL、约2.8mg/mL、约3mg/mL、约3.5mg/mL、约4mg/mL、约4.5mg/mL或约5mg/mL。In the present application, 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.
在本申请中，所述免疫检查点抑制剂的浓度可以为约1mg/mL-约10mg/mL，例如，可以为约1mg/mL-约9mg/mL，约1mg/mL-约8mg/mL，约1mg/mL-约7mg/mL，约1mg/mL-约6mg/mL，约1mg/mL-约5mg/mL，约1mg/mL-约4mg/mL，约1mg/mL-约3mg/mL或约1mg/mL-约2mg/mL；例如可以为约1mg/mL、约1.5mg/mL、约2mg/mL、约2.5mg/mL、约3mg/mL、约3.5mg/mL、约4mg/mL、约4.5mg/mL或约5mg/mL。In the present application, 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.
不欲被任何理论所限，下文中的实施例仅仅是为了阐释本申请的融合蛋白、制备方法和用途等，而不用于限制本申请发明的范围。Not to be limited by any theory, the following examples are only intended to illustrate the fusion protein, preparation method and use of the present application, and are not intended to limit the scope of the invention of the present application.
实施例Example
实施例1 本申请所述的靶向GITR的抗原结合蛋白的制备Example 1 Preparation of the antigen-binding protein targeting GITR described in this application
通过改变重链和轻链可变区的框架区的某些氨基酸残基将抗体人源化，共得到表1所示的9株人源化抗体，即9种本申请所述的分离的抗原结合蛋白，分别用3E2 1-2、3E2 1-3、3E2 1-4、3E2 2-2、3E2 2-3、3E2 2-4、3E2 3-2、3E2 3-3和3E2 3-4表示。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 .
表1. 9株人源化抗体的重链和轻链的序列Table 1. Sequences of heavy and light chains of 9 humanized antibodies
Figure PCTCN2021142299-appb-000003
Figure PCTCN2021142299-appb-000003
Figure PCTCN2021142299-appb-000004
Figure PCTCN2021142299-appb-000004
将上述人源化的VH基因和VL基因进行合成，并且将VH和人IgG1重链恒定区组成抗体的重链，VL和人kappa轻链恒定区组成抗体的轻链。各基因克隆至pcDNA4/myc-HisA的载体中，得到重链表达质粒和轻链表达质粒。分别将重链表达质粒和轻链表达质粒按照表1进行配对后瞬时转染至HEK293(ATCC，CRL-1573 TM)细胞系中用于蛋白生产。将重组表达质粒用Freestyle 293培养基稀释并加入转化所需PEI(Polyethylenimine)溶液，将每组质粒/PEI混合物分别加入细胞悬液中，放置在37℃，10％CO 2，90rpm中培养；培养5～6天后，收集瞬时表达培养上清液，通过Protein A亲和层析法，初步纯化得到9种本申请所述的分离的抗原结合蛋白，即3E2 1-2、3E2 1-3、3E2 1-4、3E2 2-2、3E2 2-3、3E2 2-4、3E2 3-2、3E2 3-3和3E2 3-4，用于以下各实施例。 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 ™ ) cell line for protein production. Dilute the recombinant expression plasmid with Freestyle 293 medium and add the PEI (Polyethylenimine) solution required for transformation. Add each group of plasmid/PEI mixtures to the cell suspension respectively, and place them at 37°C, 10% CO 2 , and cultivate at 90 rpm; culture After 5 to 6 days, the transient expression culture supernatant was collected, and 9 kinds of isolated antigen-binding proteins described in this application, namely 3E2 1-2, 3E2 1-3, 3E2, were preliminarily purified by Protein A affinity chromatography. 1-4, 3E2 2-2, 3E2 2-3, 3E2 2-4, 3E2 3-2, 3E2 3-3, and 3E2 3-4, used in the following examples.
实施例2 本申请所述药物产品抑制肿瘤生长Example 2 The drug product described in this application inhibits tumor growth
获得C57-GITR KI小鼠(即基因编辑敲入人GITR的C57背景小鼠，购自百奥赛图，向C57-GITR KI小鼠皮下接种100μL MC38细胞悬液(其中细胞悬液浓度为5×10 6个/mL，细胞悬液中细胞活率为85.87％)，得到小鼠肿瘤模型。 To obtain 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 μL 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.
然后根据表2的方式向上述小鼠肿瘤模型给药。其中小鼠PD1抗体为RMP1-14，购自Bioxcell，货号为BE0146。Then, according to the manner in Table 2, the above-mentioned mouse tumor models were administered. The mouse PD1 antibody is RMP1-14, which was purchased from Bioxcell, the product number is BE0146.
表2 各治疗组的给药情况Table 2 Administration of each treatment group
Figure PCTCN2021142299-appb-000005
Figure PCTCN2021142299-appb-000005
给药期间测定各治疗组中小鼠的体重和肿瘤体积，结果分别如图1A和图1B所示(对应治疗组1-4)。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 1A and Figure 1B, respectively (corresponding to treatment groups 1-4).
图2A-2D分别显示了人IgG1治疗组、3E2 1-4治疗组、小鼠PD1抗体治疗组和3E2 1-4+小鼠PD1抗体治疗组(即治疗组1-4)中各小鼠肿瘤体积的统计结果。各治疗组的药效分析如表3所示。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.
表3 各治疗组的药效分析Table 3 Pharmacodynamic analysis of each treatment group
Figure PCTCN2021142299-appb-000006
Figure PCTCN2021142299-appb-000006
Figure PCTCN2021142299-appb-000007
Figure PCTCN2021142299-appb-000007
上述的试验结果表明，3E2 1-4+小鼠PD1抗体治疗组的药效良好，其中2只小鼠肿瘤完全消除，可见可以有效地治疗结肠癌。The above test results show that the 3E2 1-4+ mouse PD1 antibody treatment group has a good drug effect, in which 2 mice tumors are completely eliminated, which shows that colon cancer can be effectively treated.
实施例3 本申请所述的靶向GITR的抗原结合蛋白抑制肿瘤生长Example 3 Antigen-binding protein targeting GITR described in this application inhibits tumor growth
消化，收集MC38细胞(购自ATCC)，并复苏和传代一次，PBS洗涤两次，过筛网，计数重悬至细胞悬液浓度为5×10 6个/mL，细胞悬液中细胞活率为93％。 Digested, collected MC38 cells (purchased from ATCC), recovered and passaged once, washed twice with PBS, sieved, counted and resuspended to a cell suspension concentration of 5×10 6 cells/mL, and the cell viability in the cell suspension was 93%.
获得C57-GITR KI小鼠(即基因编辑敲入人GITR的C57背景小鼠，购自百奥赛图)，向C57-GITR KI小鼠皮下接种100μL MC38细胞悬液，得到小鼠肿瘤模型。C57-GITR KI mice (that is, C57 background mice with gene editing knock-in human GITR, purchased from Bio-Ocelot) were obtained, and 100 μL of MC38 cell suspension was subcutaneously inoculated into C57-GITR KI mice to obtain a mouse tumor model.
接种后第8天，根据表4的方式向上述小鼠肿瘤模型给药。On the 8th day after inoculation, the above-mentioned mouse tumor models were administered according to the manner in Table 4.
表4 各治疗组的给药情况Table 4 Administration of each treatment group
Figure PCTCN2021142299-appb-000008
Figure PCTCN2021142299-appb-000008
给药期间测定各治疗组中小鼠的体重和肿瘤体积，结果分别如图3A和图3B所示。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.
图4A-4D分别显示了人IgG1治疗组、1mg/Kg 3E2 1-4治疗组、3mg/Kg 3E2 1-4治疗组和10mg/Kg 3E2 1-4治疗组(即治疗组1-4)中各小鼠肿瘤体积的统计结果，其中图例的数字代表老鼠的编号。各治疗组的药效分析如表5所示。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. Statistical results of tumor volume in each mouse, where the numbers in the legend represent the number of mice. The efficacy analysis of each treatment group is shown in Table 5.
表5 各治疗组的药效分析Table 5 Pharmacodynamic analysis of each treatment group
Figure PCTCN2021142299-appb-000009
Figure PCTCN2021142299-appb-000009
上述的试验结果表明，3E2 1-4可以有效地治疗结肠癌。The above test results show that 3E2 1-4 can effectively treat colon cancer.
实施例4 本申请所述药物产品抑制肿瘤生长Example 4 The drug product described in this application inhibits tumor growth
获得C57-GITR KI小鼠(即基因编辑敲入人GITR的C57背景小鼠，购自百奥赛图，向C57-GITR KI小鼠皮下接种100μL MC38细胞悬液(其中细胞悬液浓度为5×10 6个/mL，细胞悬液中细胞活率为85.87％)，得到小鼠肿瘤模型。 To obtain 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 μL 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.
然后根据表6的方式向上述小鼠肿瘤模型给药。其中小鼠PD1抗体为RMP1-14，购自Bioxcell，货号为BE0146。Then, according to the manner in Table 6, the above-mentioned mouse tumor models were administered. The mouse PD1 antibody is RMP1-14, which was purchased from Bioxcell, the product number is BE0146.
表6 各治疗组的给药情况Table 6 Administration of each treatment group
Figure PCTCN2021142299-appb-000010
Figure PCTCN2021142299-appb-000010
给药期间测定各治疗组中小鼠的体重和肿瘤体积，结果分别如图5A和图5B所示。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.
图5分别显示了人IgG1治疗组、3E2 1-4治疗组、小鼠PD1抗体治疗组和3E2 1-4+小鼠PD1抗体治疗组(即治疗组1-4)中各小鼠肿瘤体积的统计结果。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.
前述详细说明是以解释和举例的方式提供的，并非要限制所附权利要求的范围。目前本申请所列举的实施方式的多种变化对本领域普通技术人员来说是显而易见的，且保留在所附的权利要求和其等同方案的范围内。The foregoing detailed description has been presented by way of explanation and example, and is not intended to limit the scope of the appended claims. Various modifications to the embodiments presently enumerated in this application will be apparent to those of ordinary skill in the art and remain within the scope of the appended claims and their equivalents.

Claims (135)

  1. 药物产品，其包含靶向GITR的抗原结合蛋白和免疫检查点抑制剂，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中的至少一个CDR，且包含氨基酸序列如SEQ ID NO:24所示的VL中的至少一个CDR。A pharmaceutical product comprising an antigen-binding protein targeting GITR and an immune checkpoint inhibitor, wherein the antigen-binding protein comprises at least one CDR in the VH whose amino acid sequence is as shown in SEQ ID NO: 23, and which comprises an amino acid sequence such as SEQ ID NO: 23 At least one CDR in the VL shown in ID NO:24.
  2. 根据权利要求1所述的药物产品，其为药物组合物。The pharmaceutical product of claim 1, which is a pharmaceutical composition.
  3. 根据权利要求1-2中任一项所述的药物产品，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR3。The pharmaceutical product of any one of claims 1-2, wherein the antigen binding protein comprises HCDR3 in VH having an amino acid sequence as shown in SEQ ID NO:23.
  4. 根据权利要求1-3中任一项所述的药物产品，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR2。The pharmaceutical product of any one of claims 1-3, wherein the antigen binding protein comprises HCDR2 in VH having an amino acid sequence as shown in SEQ ID NO:23.
  5. 根据权利要求1-4中任一项所述的药物产品，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR1。The pharmaceutical product of any one of claims 1-4, wherein the antigen binding protein comprises HCDR1 in VH having an amino acid sequence as shown in SEQ ID NO:23.
  6. 根据权利要求1-5中任一项所述的药物产品，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR1。The pharmaceutical product of any one of claims 1-5, wherein the antigen binding protein comprises LCDR1 in VL having an amino acid sequence as shown in SEQ ID NO:24.
  7. 根据权利要求1-6中任一项所述的药物产品，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR2。The pharmaceutical product of any one of claims 1-6, wherein the antigen binding protein comprises LCDR2 in VL having an amino acid sequence as shown in SEQ ID NO:24.
  8. 根据权利要求1-7中任一项所述的药物产品，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR3。The pharmaceutical product of any one of claims 1-7, wherein the antigen binding protein comprises LCDR3 in VL having an amino acid sequence as shown in SEQ ID NO:24.
  9. 根据权利要求1-8中任一项所述的药物产品，其中所述抗原结合蛋白包含HCDR3，所述HCDR3包含SEQ ID NO:2所示的氨基酸序列。The pharmaceutical product of any one of claims 1-8, wherein the antigen binding protein comprises HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:2.
  10. 根据权利要求1-9中任一项所述的药物产品，其中所述抗原结合蛋白包含HCDR2，所述HCDR2包含SEQ ID NO:4所示的氨基酸序列。The pharmaceutical product of any one of claims 1-9, wherein the antigen binding protein comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:4.
  11. 根据权利要求1-10中任一项所述的药物产品，其中所述抗原结合蛋白包含HCDR1，所述HCDR1包含SEQ ID NO:3所示的氨基酸序列。The pharmaceutical product of any one of claims 1-10, wherein the antigen binding protein comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:3.
  12. 根据权利要求1-11中任一项所述的药物产品，其中所述抗原结合蛋白包含LCDR3，所述LCDR3包含SEQ ID NO:12所示的氨基酸序列。The pharmaceutical product of any one of claims 1-11, wherein the antigen binding protein comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:12.
  13. 根据权利要求1-12中任一项所述的药物产品，其中所述抗原结合蛋白包含LCDR2，所述LCDR2包含SEQ ID NO:11所示的氨基酸序列。The pharmaceutical product of any one of claims 1-12, wherein the antigen binding protein comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:11.
  14. 根据权利要求1-13中任一项所述的药物产品，其中所述抗原结合蛋白包含LCDR1，所述LCDR1包含SEQ ID NO:10所示的氨基酸序列。The pharmaceutical product of any one of claims 1-13, wherein the antigen binding protein comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:10.
  15. 根据权利要求1-14中任一项所述的药物产品，其中所述抗原结合蛋白包括抗体或其抗原结合片段。The pharmaceutical product of any one of claims 1-14, wherein the antigen-binding protein comprises an antibody or antigen-binding fragment thereof.
  16. 根据权利要求15所述的药物产品，其中所述抗原结合片段包括Fab，Fab’，F(ab)2、Fv 片段、F(ab’) 2，scFv，di-scFv和/或dAb。 The pharmaceutical product of claim 15, wherein the antigen binding fragment comprises Fab, Fab', F(ab)2, Fv fragment, F(ab') 2 , scFv, di-scFv and/or dAb.
  17. 根据权利要求1-16中任一项所述的药物产品，其中所述抗体为人源化抗体。The pharmaceutical product of any one of claims 1-16, wherein the antibody is a humanized antibody.
  18. 根据权利要求1-17中任一项所述的药物产品，其中所述VL包括框架区L-FR1，L-FR2，L-FR3，和L-FR4。The pharmaceutical product of any one of claims 1-17, wherein the VL comprises framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
  19. 根据权利要求18所述的药物产品，其中所述L-FR1的C末端与所述LCDR1的N末端直接或间接相连，且所述L-FR1包含SEQ ID NO:39所示的氨基酸序列。The pharmaceutical product of claim 18, wherein 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 shown in SEQ ID NO:39.
  20. 根据权利要求18-19中任一项所述的药物产品，其中所述L-FR1包含SEQ ID NO:13、30、34中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 18-19, wherein the L-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 13, 30, 34.
  21. 根据权利要求18-20中任一项所述的药物产品，其中所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包含SEQ ID NO:40所示的氨基酸序列。The pharmaceutical product of any one of claims 18-20, wherein 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 .
  22. 根据权利要求18-21中任一项所述的药物产品，其中所述L-FR2包含SEQ ID NO:14、31中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 18-21, wherein the L-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 14, 31.
  23. 根据权利要求18-22中任一项所述的药物产品，其中所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO:41所示的氨基酸序列。The pharmaceutical product of any one of claims 18-22, wherein 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 .
  24. 根据权利要求18-23中任一项所述的药物产品，其中所述L-FR3包含SEQ ID NO:15、32中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 18-23, wherein the L-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 15, 32.
  25. 根据权利要求18-24中任一项所述的药物产品，其中所述L-FR4的N末端与所述LCDR3的C末端相连，且所述L-FR4包含SEQ ID NO:42所示的氨基酸序列。The pharmaceutical product of any one of claims 18-24, wherein the N-terminus of the L-FR4 is linked to the C-terminus of the LCDR3, and the L-FR4 comprises the amino acid set forth in SEQ ID NO:42 sequence.
  26. 根据权利要求18-25中任一项所述的药物产品，其中所述L-FR4包含SEQ ID NO:16、33中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 18-25, wherein the L-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 16, 33.
  27. 根据权利要求1-26中任一项所述的药物产品，其中所述VL包含SEQ ID NO:24所示的氨基酸序列。The pharmaceutical product of any one of claims 1-26, wherein the VL comprises the amino acid sequence set forth in SEQ ID NO:24.
  28. 根据权利要求1-27中任一项所述的药物产品，其中所述VL包含SEQ ID NO:9、20-22中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 1-27, wherein the VL comprises the amino acid sequence set forth in any one of SEQ ID NOs: 9, 20-22.
  29. 根据权利要求1-28中任一项所述的药物产品，其中所述抗原结合蛋白包括抗体轻链恒定区，且所述抗体轻链恒定区包括人Igκ恒定区。The pharmaceutical product of any one of claims 1-28, wherein the antigen binding protein comprises an antibody light chain constant region, and the antibody light chain constant region comprises a human IgK constant region.
  30. 根据权利要求29所述的药物产品，其中所述抗体轻链恒定区包含SEQ ID NO:52所示的氨基酸序列。The pharmaceutical product of claim 29, wherein the antibody light chain constant region comprises the amino acid sequence shown in SEQ ID NO:52.
  31. 根据权利要求1-30中任一项所述的药物产品，其中所述抗原结合蛋白包含抗体轻链LC，且所述LC包含SEQ ID NO:50、46-48中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 1-30, wherein the antigen binding protein comprises an antibody light chain LC, and the LC comprises the amino acids set forth in any of SEQ ID NOs: 50, 46-48 sequence.
  32. 根据权利要求1-31中任一项所述的药物产品，其中所述VH包括框架区H-FR1，H- FR2，H-FR3，和H-FR4。The pharmaceutical product of any one of claims 1-31, wherein the VH comprises the framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
  33. 根据权利要求32所述的药物产品，其中所述H-FR1的C末端与所述HCDR1的N末端直接或间接相连，且所述H-FR1包含SEQ ID NO:35所示的氨基酸序列。The pharmaceutical product of claim 32, wherein 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 shown in SEQ ID NO:35.
  34. 根据权利要求32-33中任一项所述的药物产品，其中所述H-FR1包含SEQ ID NO:5、25、29中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 32-33, wherein the H-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 5, 25, 29.
  35. 根据权利要求32-34中任一项所述的药物产品，其中所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO:36所示的氨基酸序列。The pharmaceutical product of any one of claims 32-34, wherein 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 .
  36. 根据权利要求32-35中任一项所述的药物产品，其中所述H-FR2包含SEQ ID NO:6、26中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 32-35, wherein the H-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 6, 26.
  37. 根据权利要求32-36中任一项所述的药物产品，其中所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO:37所示的氨基酸序列。The pharmaceutical product of any one of claims 32-36, wherein 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 .
  38. 根据权利要求32-37中任一项所述的药物产品，其中所述H-FR3包含SEQ ID NO:7、27中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 32-37, wherein the H-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 7, 27.
  39. 根据权利要求32-38中任一项所述的药物产品，其中所述H-FR4的N末端与所述HCDR3的C末端相连，且所述H-FR4包含SEQ ID NO:38所示的氨基酸序列。The pharmaceutical product of any one of claims 32-38, wherein the N-terminus of the H-FR4 is linked to the C-terminus of the HCDR3, and the H-FR4 comprises the amino acid set forth in SEQ ID NO:38 sequence.
  40. 根据权利要求32-39中任一项所述的药物产品，其中所述H-FR4包含SEQ ID NO:8、28中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 32-39, wherein the H-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 8, 28.
  41. 根据权利要求32-40中任一项所述的药物产品，其中所述VH包含SEQ ID NO:23所示的氨基酸序列。The pharmaceutical product of any one of claims 32-40, wherein the VH comprises the amino acid sequence set forth in SEQ ID NO:23.
  42. 根据权利要求32-41中任一项所述的药物产品，其中所述VH包含SEQ ID NO:1、17-19中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 32-41, wherein the VH comprises the amino acid sequence set forth in any one of SEQ ID NOs: 1, 17-19.
  43. 根据权利要求1-42中任一项所述的药物产品，其中所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG重链恒定区。The pharmaceutical product of any one of claims 1-42, wherein 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.
  44. 根据权利要求1-43中任一项所述的药物产品，其中所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG1重链恒定区。The pharmaceutical product of any one of claims 1-43, wherein 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.
  45. 根据权利要求43-44中任一项所述的药物产品，其中所述抗体重链恒定区包含SEQ ID NO:51所示的氨基酸序列。The pharmaceutical product of any one of claims 43-44, wherein the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:51.
  46. 根据权利要求1-45中任一项所述的药物产品，其中所述抗原结合蛋白包含抗体重链HC，且所述HC包含SEQ ID NO:49、43-45中任一项所示的氨基酸序列。The pharmaceutical product of any one of claims 1-45, wherein the antigen binding protein comprises an antibody heavy chain HC, and the HC comprises the amino acids set forth in any of SEQ ID NOs: 49, 43-45 sequence.
  47. 根据权利要求1-46中任一项所述的药物产品，其中所述免疫检查点抑制剂阻碍PD-1和PD-L1的相互作用。The pharmaceutical product of any one of claims 1-46, wherein the immune checkpoint inhibitor blocks the interaction of PD-1 and PD-L1.
  48. 根据权利要求1-47中任一项所述的药物产品，其中所述免疫检查点抑制剂包含PD-1抑制剂。The pharmaceutical product of any one of claims 1-47, wherein the immune checkpoint inhibitor comprises a PD-1 inhibitor.
  49. 根据权利要求1-48中任一项所述的药物产品，其中所述免疫检查点抑制剂包含PD-1抗体或其抗原结合片段。The pharmaceutical product of any one of claims 1-48, wherein the immune checkpoint inhibitor comprises a PD-1 antibody or antigen-binding fragment thereof.
  50. 根据权利要求49所述的药物产品，其中所述PD-1抗体或抗原结合片段包含HCDR3，其中所述PD-1抗体或抗原结合片段的HCDR3包含RMP1-14的HCDR3。The pharmaceutical product of claim 49, wherein 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.
  51. 根据权利要求49-50中任一项所述的药物产品，其中所述PD-1抗体或抗原结合片段包含HCDR2，其中所述PD-1抗体或抗原结合片段的HCDR2包含RMP1-14的HCDR2。The pharmaceutical product of any one of claims 49-50, wherein 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.
  52. 根据权利要求49-51中任一项所述的药物产品，其中所述PD-1抗体或抗原结合片段包含HCDR1，其中所述PD-1抗体或抗原结合片段的HCDR1包含RMP1-14的HCDR1。The pharmaceutical product of any one of claims 49-51, wherein 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.
  53. 根据权利要求49-52中任一项所述的药物产品，其中所述PD-1抗体或抗原结合片段包含LCDR3，其中所述PD-1抗体或抗原结合片段的LCDR3包含RMP1-14的LCDR3。The pharmaceutical product of any one of claims 49-52, wherein 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.
  54. 根据权利要求49-53中任一项所述的药物产品，其中所述PD-1抗体或抗原结合片段包含LCDR2，其中所述PD-1抗体或抗原结合片段的LCDR2包含RMP1-14的LCDR2。The pharmaceutical product of any one of claims 49-53, wherein the PD-1 antibody or antigen-binding fragment comprises LCDR2, wherein the LCDR2 of the PD-1 antibody or antigen-binding fragment comprises LCDR2 of RMP1-14.
  55. 根据权利要求49-54中任一项所述的药物产品，其中所述PD-1抗体或抗原结合片段包含LCDR1，其中所述PD-1抗体或抗原结合片段的所述LCDR1包含RMP1-14的LCDR1。The drug product of any one of claims 49-54, wherein the PD-1 antibody or antigen-binding fragment comprises LCDR1, wherein the LCDR1 of the PD-1 antibody or antigen-binding fragment comprises RMP1-14 LCDR1.
  56. 根据权利要求49-55中任一项所述的药物产品，其中所述PD-1抗体或抗原结合片段包含VH，所述VH包含RMP1-14的VH。The drug product of any one of claims 49-55, wherein the PD-1 antibody or antigen-binding fragment comprises a VH comprising the VH of RMP1-14.
  57. 根据权利要求49-56中任一项所述的药物产品，其中所述PD-1抗体或抗原结合片段包含VL，所述VL包含RMP1-14的VL。The pharmaceutical product of any one of claims 49-56, wherein the PD-1 antibody or antigen-binding fragment comprises a VL comprising the VL of RMP1-14.
  58. 根据权利要求1-57中任一项所述的药物产品，其中所述靶向GITR的抗原结合蛋白和所述免疫检查点抑制剂在所述药物产品中相互不混合。The drug product of any one of claims 1-57, wherein the GITR-targeting antigen binding protein and the immune checkpoint inhibitor are not mixed with each other in the drug product.
  59. 根据权利要求1-58中任一项所述的药物产品，其中所述靶向GITR的抗原结合蛋白和所述免疫检查点抑制剂在所述药物产品中分别存在于不同的容器中。The pharmaceutical product of any one of claims 1-58, wherein the GITR-targeting antigen binding protein and the immune checkpoint inhibitor are present in separate containers in the pharmaceutical product.
  60. 权利要求1-59中任一项所述的药物产品在制备预防、缓解或治疗癌症的药物中的用途。Use of the pharmaceutical product of any one of claims 1-59 in the manufacture of a medicament for preventing, relieving or treating cancer.
  61. 根据权利要求60所述的用途，其中所述癌症包括GITR阳性的癌症；和/或，所述癌症包括PD-1阳性的癌症。The use of claim 60, wherein the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
  62. 根据权利要求60-61中任一项所述的用途，其中所述癌症包括实体瘤。The use of any one of claims 60-61, wherein the cancer comprises a solid tumor.
  63. 根据权利要求60-62中任一项所述的用途，其中所述癌症选自下组：结肠癌、直肠癌和乳腺癌。The use of any one of claims 60-62, wherein the cancer is selected from the group consisting of colon cancer, rectal cancer and breast cancer.
  64. 靶向GITR的抗原结合蛋白和免疫检查点拮抗剂在制备预防、缓解或治疗癌症的药物中 的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中的至少一个CDR，且包含氨基酸序列如SEQ ID NO:24所示的VL中的至少一个CDR。Antigen-binding protein targeting GITR and use of immune checkpoint antagonists in the preparation of drugs for preventing, relieving or treating cancer, wherein the antigen-binding protein comprises at least one of the VHs whose amino acid sequence is shown in SEQ ID NO: 23 CDRs and comprising at least one CDR in VL whose amino acid sequence is set forth in SEQ ID NO:24.
  65. 根据权利要求64所述的用途，其中所述癌症包括GITR阳性的癌症；和/或，所述癌症包括PD-1阳性的癌症。The use of claim 64, wherein the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
  66. 根据权利要求64-65中任一项所述的用途，其中所述癌症包括实体瘤。The use of any one of claims 64-65, wherein the cancer comprises a solid tumor.
  67. 根据权利要求64-66中任一项所述的用途，其中所述癌症选自下组：结肠癌、直肠癌和乳腺癌。The use of any one of claims 64-66, wherein the cancer is selected from the group consisting of colon cancer, rectal cancer and breast cancer.
  68. 根据权利要求64-67中任一项所述的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR3。The use of any one of claims 64-67, wherein the antigen binding protein comprises HCDR3 in VH having an amino acid sequence as set forth in SEQ ID NO:23.
  69. 根据权利要求64-68中任一项所述的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR2。The use of any one of claims 64-68, wherein the antigen binding protein comprises HCDR2 in VH having an amino acid sequence as set forth in SEQ ID NO:23.
  70. 根据权利要求64-69中任一项所述的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:23所示的VH中HCDR1。The use of any one of claims 64-69, wherein the antigen binding protein comprises HCDR1 in VH having an amino acid sequence as set forth in SEQ ID NO:23.
  71. 根据权利要求64-70中任一项所述的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR1。The use of any one of claims 64-70, wherein the antigen binding protein comprises LCDR1 in VL having an amino acid sequence as set forth in SEQ ID NO:24.
  72. 根据权利要求64-71中任一项所述的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR2。The use of any one of claims 64-71, wherein the antigen binding protein comprises LCDR2 in VL having an amino acid sequence as set forth in SEQ ID NO:24.
  73. 根据权利要求64-72中任一项所述的用途，其中所述抗原结合蛋白包含氨基酸序列如SEQ ID NO:24所示的VL中LCDR3。The use of any one of claims 64-72, wherein the antigen binding protein comprises LCDR3 in VL having an amino acid sequence as set forth in SEQ ID NO:24.
  74. 根据权利要求64-73中任一项所述的用途，其中所述抗原结合蛋白包含HCDR3，所述HCDR3包含SEQ ID NO:2所示的氨基酸序列。The use of any one of claims 64-73, wherein the antigen binding protein comprises HCDR3 comprising the amino acid sequence set forth in SEQ ID NO:2.
  75. 根据权利要求64-74中任一项所述的用途，其中所述抗原结合蛋白包含HCDR2，所述HCDR2包含SEQ ID NO:4所示的氨基酸序列。The use of any one of claims 64-74, wherein the antigen binding protein comprises HCDR2 comprising the amino acid sequence set forth in SEQ ID NO:4.
  76. 根据权利要求64-75中任一项所述的用途，其中所述抗原结合蛋白包含HCDR1，所述HCDR1包含SEQ ID NO:3所示的氨基酸序列。The use of any one of claims 64-75, wherein the antigen binding protein comprises HCDR1 comprising the amino acid sequence set forth in SEQ ID NO:3.
  77. 根据权利要求64-76中任一项所述的用途，其中所述抗原结合蛋白包含LCDR3，所述LCDR3包含SEQ ID NO:12所示的氨基酸序列。The use of any one of claims 64-76, wherein the antigen binding protein comprises LCDR3 comprising the amino acid sequence set forth in SEQ ID NO:12.
  78. 根据权利要求64-77中任一项所述的用途，其中所述抗原结合蛋白包含LCDR2，所述LCDR2包含SEQ ID NO:11所示的氨基酸序列。The use of any one of claims 64-77, wherein the antigen binding protein comprises LCDR2 comprising the amino acid sequence set forth in SEQ ID NO:11.
  79. 根据权利要求64-78中任一项所述的用途，其中所述抗原结合蛋白包含LCDR1，所述LCDR1包含SEQ ID NO:10所示的氨基酸序列。The use of any one of claims 64-78, wherein the antigen binding protein comprises LCDR1 comprising the amino acid sequence set forth in SEQ ID NO:10.
  80. 根据权利要求64-79中任一项所述的用途，其中所述抗原结合蛋白包括抗体或其抗原结合片段。The use of any one of claims 64-79, wherein the antigen-binding protein comprises an antibody or antigen-binding fragment thereof.
  81. 根据权利要求80所述的用途，其中所述抗原结合片段包括Fab，Fab’，F(ab)2、Fv片段、F(ab’)2，scFv，di-scFv和/或dAb。The use of claim 80, wherein the antigen-binding fragment comprises Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.
  82. 根据权利要求80-81中任一项所述的用途，其中所述抗体为人源化抗体。The use of any one of claims 80-81, wherein the antibody is a humanized antibody.
  83. 根据权利要求64-82中任一项所述的用途，其中所述VL包括框架区L-FR1，L-FR2，L-FR3，和L-FR4。The use of any one of claims 64-82, wherein the VL comprises framework regions L-FR1, L-FR2, L-FR3, and L-FR4.
  84. 根据权利要求83所述的用途，其中所述L-FR1的C末端与所述LCDR1的N末端直接或间接相连，且所述L-FR1包含SEQ ID NO:39所示的氨基酸序列。The use of claim 83, wherein 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 shown in SEQ ID NO:39.
  85. 根据权利要求83-84中任一项所述的用途，其中所述L-FR1包含SEQ ID NO:13、30、34中任一项所示的氨基酸序列。The use of any one of claims 83-84, wherein the L-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 13, 30, 34.
  86. 根据权利要求83-85中任一项所述的用途，其中所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包含SEQ ID NO:40所示的氨基酸序列。The use of any one of claims 83-85, wherein 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.
  87. 根据权利要求83-86中任一项所述的用途，其中所述L-FR2包含SEQ ID NO:14、31中任一项所示的氨基酸序列。The use of any one of claims 83-86, wherein the L-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 14, 31.
  88. 根据权利要求83-87中任一项所述的用途，其中所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO:41所示的氨基酸序列。The use of any one of claims 83-87, wherein 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.
  89. 根据权利要求83-88中任一项所述的用途，其中所述L-FR3包含SEQ ID NO:15、32中任一项所示的氨基酸序列。The use of any one of claims 83-88, wherein the L-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 15, 32.
  90. 根据权利要求83-89中任一项所述的用途，其中所述L-FR4的N末端与所述LCDR3的C末端相连，且所述L-FR4包含SEQ ID NO:42所示的氨基酸序列。The use of any one of claims 83-89, wherein 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 shown in SEQ ID NO:42 .
  91. 根据权利要求83-90中任一项所述的用途，其中所述L-FR4包含SEQ ID NO:16、33中任一项所示的氨基酸序列。The use of any one of claims 83-90, wherein the L-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 16, 33.
  92. 根据权利要求64-91中任一项所述的用途，其中所述VL包含SEQ ID NO:24所示的氨基酸序列。The use of any one of claims 64-91, wherein the VL comprises the amino acid sequence set forth in SEQ ID NO:24.
  93. 根据权利要求64-92中任一项所述的用途，其中所述VL包含SEQ ID NO:9、20-22中任一项所示的氨基酸序列。The use of any one of claims 64-92, wherein the VL comprises the amino acid sequence set forth in any one of SEQ ID NOs: 9, 20-22.
  94. 根据权利要求64-93中任一项所述的用途，其中所述抗原结合蛋白包括抗体轻链恒定区，且所述抗体轻链恒定区包括人Igκ恒定区。The use of any one of claims 64-93, wherein the antigen binding protein comprises an antibody light chain constant region, and the antibody light chain constant region comprises a human IgK constant region.
  95. 根据权利要求94所述的用途，其中所述抗体轻链恒定区包含SEQ ID NO:52所示的氨基酸序列。The use of claim 94, wherein the antibody light chain constant region comprises the amino acid sequence shown in SEQ ID NO:52.
  96. 根据权利要求64-95中任一项所述的用途，其中所述抗原结合蛋白包含抗体轻链LC，且所述LC包含SEQ ID NO:50、46-48中任一项所示的氨基酸序列。The use of any one of claims 64-95, wherein the antigen binding protein comprises an antibody light chain LC, and the LC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 50, 46-48 .
  97. 根据权利要求64-96中任一项所述的用途，其中所述VH包括框架区H-FR1，H-FR2，H-FR3，和H-FR4。The use of any one of claims 64-96, wherein the VH comprises framework regions H-FR1, H-FR2, H-FR3, and H-FR4.
  98. 根据权利要求97所述的用途，其中所述H-FR1的C末端与所述HCDR1的N末端直接或间接相连，且所述H-FR1包含SEQ ID NO:35所示的氨基酸序列。The use of claim 97, wherein 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 shown in SEQ ID NO:35.
  99. 根据权利要求97-98中任一项所述的用途，其中所述H-FR1包含SEQ ID NO:5、25、29中任一项所示的氨基酸序列。The use of any one of claims 97-98, wherein the H-FR1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 5, 25, 29.
  100. 根据权利要求97-99中任一项所述的用途，其中所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO:36所示的氨基酸序列。The use of any one of claims 97-99, wherein 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.
  101. 根据权利要求97-100中任一项所述的用途，其中所述H-FR2包含SEQ ID NO:6、26中任一项所示的氨基酸序列。The use of any one of claims 97-100, wherein the H-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 6, 26.
  102. 根据权利要求97-101中任一项所述的用途，其中所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO:37所示的氨基酸序列。The use of any one of claims 97-101, wherein 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.
  103. 根据权利要求97-102中任一项所述的用途，其中所述H-FR3包含SEQ ID NO:7、27中任一项所示的氨基酸序列。The use of any one of claims 97-102, wherein the H-FR3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 7, 27.
  104. 根据权利要求97-103中任一项所述的用途，其中所述H-FR4的N末端与所述HCDR3的C末端相连，且所述H-FR4包含SEQ ID NO:38所示的氨基酸序列。The use of any one of claims 97-103, wherein 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 .
  105. 根据权利要求97-104中任一项所述的用途，其中所述H-FR4包含SEQ ID NO:8、28中任一项所示的氨基酸序列。The use of any one of claims 97-104, wherein the H-FR4 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 8, 28.
  106. 根据权利要求64-105中任一项所述的用途，其中所述VH包含SEQ ID NO:23所示的氨基酸序列。The use of any one of claims 64-105, wherein the VH comprises the amino acid sequence set forth in SEQ ID NO:23.
  107. 根据权利要求64-106中任一项所述的用途，其中所述VH包含SEQ ID NO:1、17-19中任一项所示的氨基酸序列。The use of any one of claims 64-106, wherein the VH comprises the amino acid sequence set forth in any one of SEQ ID NOs: 1, 17-19.
  108. 根据权利要求64-107中任一项所述的用途，其中所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG重链恒定区。The use of any one of claims 64-107, wherein 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.
  109. 根据权利要求64-108中任一项所述的用途，其中所述抗原结合蛋白包括抗体重链恒定区，且所述抗体重链恒定区源自人IgG1重链恒定区。The use of any one of claims 64-108, wherein 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.
  110. 根据权利要求64-109中任一项所述的用途，其中所述抗体重链恒定区包含SEQ ID NO:51所示的氨基酸序列。The use of any one of claims 64-109, wherein the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:51.
  111. 根据权利要求64-110中任一项所述的用途，其中所述抗原结合蛋白包含抗体重链 HC，且所述HC包含SEQ ID NO:49、43-45中任一项所示的氨基酸序列。The use of any one of claims 64-110, wherein the antigen binding protein comprises an antibody heavy chain HC, and the HC comprises the amino acid sequence set forth in any one of SEQ ID NOs: 49, 43-45 .
  112. 根据权利要求64-111中任一项所述的用途，其中所述免疫检查点抑制剂阻碍PD-1和PD-L1的相互作用。The use of any one of claims 64-111, wherein the immune checkpoint inhibitor blocks the interaction of PD-1 and PD-L1.
  113. 根据权利要求64-112中任一项所述的用途，其中所述免疫检查点抑制剂包含PD-1抑制剂。The use of any one of claims 64-112, wherein the immune checkpoint inhibitor comprises a PD-1 inhibitor.
  114. 根据权利要求64-113中任一项所述的用途，其中所述免疫检查点抑制剂包含PD-1抗体或其抗原结合片段。The use of any one of claims 64-113, wherein the immune checkpoint inhibitor comprises a PD-1 antibody or antigen-binding fragment thereof.
  115. 根据权利要求114所述的用途，其中所述PD-1抗体或抗原结合片段包含HCDR3，其中所述PD-1抗体或抗原结合片段的HCDR3包含RMP1-14的HCDR3。The use of claim 114, wherein 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.
  116. 根据权利要求114-115中任一项所述的用途，其中所述PD-1抗体或抗原结合片段包含HCDR2，其中所述PD-1抗体或抗原结合片段的HCDR2包含RMP1-14的HCDR2。The use of any one of claims 114-115, wherein 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.
  117. 根据权利要求114-116中任一项所述的用途，其中所述PD-1抗体或抗原结合片段包含HCDR1，其中所述PD-1抗体或抗原结合片段的HCDR1包含RMP1-14的HCDR1。The use of any one of claims 114-116, wherein 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.
  118. 根据权利要求114-117中任一项所述的用途，其中所述PD-1抗体或抗原结合片段包含LCDR3，其中所述PD-1抗体或抗原结合片段的LCDR3包含RMP1-14的LCDR3。The use of any one of claims 114-117, wherein 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.
  119. 根据权利要求114-118中任一项所述的用途，其中所述PD-1抗体或抗原结合片段包含LCDR2，其中所述PD-1抗体或抗原结合片段的LCDR2包含RMP1-14的LCDR2。The use of any one of claims 114-118, wherein the PD-1 antibody or antigen-binding fragment comprises LCDR2, wherein the LCDR2 of the PD-1 antibody or antigen-binding fragment comprises LCDR2 of RMP1-14.
  120. 根据权利要求114-119中任一项所述的用途，其中所述PD-1抗体或抗原结合片段包含LCDR1，其中所述PD-1抗体或抗原结合片段的所述LCDR1包含RMP1-14的LCDR1。The use of any one of claims 114-119, wherein 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 .
  121. 根据权利要求114-120中任一项所述的用途，其中所述PD-1抗体或抗原结合片段包含VH，所述VH包含RMP1-14的VH。The use of any one of claims 114-120, wherein the PD-1 antibody or antigen-binding fragment comprises a VH comprising the VH of RMP1-14.
  122. 根据权利要求114-121中任一项所述的用途，其中所述PD-1抗体或抗原结合片段包含VL，所述VL包含RMP1-14的VL。The use of any one of claims 114-121, wherein the PD-1 antibody or antigen-binding fragment comprises a VL comprising the VL of RMP1-14.
  123. 一种预防、缓解或治疗癌症的方法，其包括以下步骤：向有需要的受试者施用权利要求1-59中任一项所述的靶向GITR的抗原结合蛋白，和/或，向有需要的受试者施用权利要求1-59中任一项所述的免疫检查点抑制剂。A method of preventing, relieving or treating cancer, comprising the steps of: administering the GITR-targeting antigen binding protein of any one of claims 1-59 to a subject in need, and/or, to a subject in need A subject in need thereof is administered the immune checkpoint inhibitor of any one of claims 1-59.
  124. 根据权利要求123所述的方法，其包括以下步骤：向有需要的受试者施用权利要求1-59中任一项所述的药物产品。The method of claim 123, comprising the step of administering the pharmaceutical product of any one of claims 1-59 to a subject in need thereof.
  125. 根据权利要求123-124中任一项所述的方法，其中所述癌症包括GITR阳性的癌症；和/或，所述癌症包括PD-1阳性的癌症。The method of any one of claims 123-124, wherein the cancer comprises a GITR positive cancer; and/or the cancer comprises a PD-1 positive cancer.
  126. 根据权利要求123-125中任一项所述的方法，其中所述癌症包括实体瘤。The method of any one of claims 123-125, wherein the cancer comprises a solid tumor.
  127. 根据权利要求123-126中任一项所述的方法，所述癌症选自下组：结肠癌、直肠癌和乳腺癌。The method of any one of claims 123-126, wherein the cancer is selected from the group consisting of colon cancer, rectal cancer, and breast cancer.
  128. 根据权利要求123-127中任一项所述的方法，其中所述抗原结合蛋白的给药剂量为1mg/Kg-10mg/Kg。The method of any one of claims 123-127, wherein the antigen binding protein is administered at a dose of 1 mg/Kg to 10 mg/Kg.
  129. 根据权利要求123-128中任一项所述的方法，其中所述免疫检查点抑制剂的给药剂量为1mg/Kg-10mg/Kg。The method of any one of claims 123-128, wherein the immune checkpoint inhibitor is administered at a dose of 1 mg/Kg to 10 mg/Kg.
  130. 根据权利要求123-129中任一项所述的方法，其中所述抗原结合蛋白的给药频率为每周2次。The method of any one of claims 123-129, wherein the frequency of administration of the antigen binding protein is twice a week.
  131. 根据权利要求123-130中任一项所述的方法，其中所述免疫检查点抑制剂的给药频率为每周2次。The method of any one of claims 123-130, wherein the frequency of administration of the immune checkpoint inhibitor is twice a week.
  132. 根据权利要求123-131中任一项所述的方法，其中所述抗原结合蛋白的给药方式为注射。The method of any one of claims 123-131, wherein the antigen binding protein is administered by injection.
  133. 根据权利要求123-132中任一项所述的方法，其中所述免疫检查点抑制剂的给药方式为注射。The method of any one of claims 123-132, wherein the immune checkpoint inhibitor is administered by injection.
  134. 根据权利要求123-133中任一项所述的方法，其中所述抗原结合蛋白的浓度为1mg/mL-10mg/mL。The method of any one of claims 123-133, wherein the antigen binding protein is at a concentration of 1 mg/mL to 10 mg/mL.
  135. 根据权利要求123-134中任一项所述的方法，其中所述免疫检查点抑制剂的浓度为1mg/mL-10mg/mL。The method of any one of claims 123-134, wherein the concentration of the immune checkpoint inhibitor is 1 mg/mL to 10 mg/mL.
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