WO2023001118A1 - Application of anti-ox40 antibody in combined drug - Google Patents

Application of anti-ox40 antibody in combined drug Download PDF

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WO2023001118A1
WO2023001118A1 PCT/CN2022/106326 CN2022106326W WO2023001118A1 WO 2023001118 A1 WO2023001118 A1 WO 2023001118A1 CN 2022106326 W CN2022106326 W CN 2022106326W WO 2023001118 A1 WO2023001118 A1 WO 2023001118A1
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antibody
seq
sequence
antigen
amino acid
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PCT/CN2022/106326
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French (fr)
Chinese (zh)
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梁世忠
符子艺
宋述强
俞金泉
李胜峰
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百奥泰生物制药股份有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • 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
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A61K2039/507—Comprising a combination of two or more separate 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
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule

Definitions

  • the invention belongs to the field of biomedicine, in particular to the application of anti-OX40 antibody in combined medicine.
  • T cells in the immune system There are two types of checkpoints in the immune system, one is inhibitory, such as PD-1, and the other is activating, such as OX40.
  • Adequate activation of T cells in the immune system requires two levels of signaling. The first signal is generated by T cell antigen receptor recognition of antigen, and the activation signal is transferred into the cell through the CD3 molecule; while the second signal is called costimulatory signal, which is generated by costimulatory molecules on the surface of antigen-presenting cells or target cells and activated Produced by the interaction of co-stimulatory molecule receptors on the surface of T cells.
  • Costimulatory signals promote the proliferation and differentiation of antigen-specific T cells into effector T cells (Lindsay K et al. Immunity 2016,44(5):1005-1019).
  • OX40 also known as TNFRSF4, ACT35, CD134, etc., belongs to the tumor necrosis factor receptor superfamily (TNFRSF), which is a type I transmembrane glycoprotein.
  • TNFRSF tumor necrosis factor receptor superfamily
  • OX40 is not expressed on resting T cells, but on activated CD4 + T cells, CD8 + T cells, NK cells and NKT cells (Paterson DJ et al. Mol. Immunol. 1987; 24:1281-1290). After T cells are activated by antigens, they will highly express OX40 molecules within 1-3 days. The activation of OX40 signal will further enhance the activation signal of T cells to enhance the response of the immune system (Gramaglia I et al. J. Immunol. 2000; 165:3043-3050).
  • anti-OX40 antibodies mainly activate the immune system and inhibit tumors through the following three cellular physiological mechanisms.
  • One is to directly activate CD4 + and CD8 + effector T cells, promote their proliferation and survival, and secrete related inflammatory factors; the other is to weaken the suppression of the immune system by inhibiting the signal and activity of Treg
  • the third is to deplete Treg cells through ADCC or ADCP, etc., to reduce their suppression of effector T cells (J. Willoughby et al. Molecular Immunology 83, 2017, 13-22).
  • OX40 is a very potential activating target in tumor immunotherapy, which can provide a new means for tumor immunotherapy.
  • the invention discloses a method or application of anti-OX40 antibody or antigen-binding fragment for combined treatment of tumor or cancer.
  • the method or use comprises: administering to a patient in need thereof an effective amount of an anti-OX40 antibody or antigen-binding fragment and another therapeutic agent.
  • the present invention discloses the use of an anti-OX40 antibody (such as antibody M or M-KF) or an antigen-binding fragment and another therapeutic agent in the preparation of a medicament for treating tumor or cancer.
  • the present invention discloses the use of an anti-OX40 antibody (such as Antibody M or Antibody M-KF) or an antigen-binding fragment for the preparation of a medicament for use in combination with another therapeutic agent.
  • an anti-OX40 antibody such as Antibody M or Antibody M-KF
  • an antigen-binding fragment for the preparation of a medicament for use in combination with another therapeutic agent.
  • the present invention discloses the use of an anti-OX40 antibody (such as antibody M or antibody M-KF) or an antigen-binding fragment in combination with another therapeutic agent in the treatment of tumor or cancer.
  • an anti-OX40 antibody such as antibody M or antibody M-KF
  • an antigen-binding fragment in combination with another therapeutic agent in the treatment of tumor or cancer.
  • the present invention also discloses a kit, which comprises an anti-OX40 antibody or an antigen-binding fragment (or preparation), another therapeutic agent (or preparation), and an anti-OX40 antibody for guiding patients in need thereof. or instructions for an antigen-binding fragment (or formulation) and another therapeutic agent (or formulation).
  • the present invention also discloses a kit comprising a composition (or preparation) of an anti-OX40 antibody or an antigen-binding fragment and another therapeutic agent and a method for guiding administration of an anti-OX40 antibody to a patient in need thereof. or instructions for a composition (or formulation) of an antigen-binding fragment and another therapeutic agent.
  • the present invention also discloses a pharmaceutical composition suitable for injection, such as a bolus injection type pharmaceutical composition or an infusion (drip) type pharmaceutical composition, comprising an anti-OX40 antibody or an antigen-binding fragment and another therapeutic agent.
  • a pharmaceutical composition suitable for injection such as a bolus injection type pharmaceutical composition or an infusion (drip) type pharmaceutical composition, comprising an anti-OX40 antibody or an antigen-binding fragment and another therapeutic agent.
  • the pharmaceutical composition comprises at least 0.1% of an anti-OX40 antibody or antigen-binding fragment and 0.1% of another therapeutic agent.
  • the percentage of antibody or antigen-binding fragment and another therapeutic agent can vary and can be between about 2% and about 90% by weight of a given dosage form.
  • the amount of anti-OX40 antibody or antigen-binding fragment and another therapeutic agent in such therapeutically useful pharmaceutical compositions can be an effective amount for administration.
  • the present invention also discloses a preparation method of the above-mentioned pharmaceutical composition: separately combining the anti-OX40 antibody or antigen-binding fragment described herein and another therapeutic agent (or the anti-OX40 antibody or antigen-binding fragment and another therapeutic agent) Composition) mixed with a pharmaceutically acceptable carrier suitable for injection (such as water for injection, physiological saline, etc.).
  • a pharmaceutically acceptable carrier suitable for injection such as water for injection, physiological saline, etc.
  • an anti-OX40 antibody or antigen-binding fragment is used in combination with another therapeutic agent to treat a tumor or cancer.
  • anti-OX40 antibody or antigen-binding fragment (or preparation) and another therapeutic agent (or preparation) are used in the treatment of tumor or cancer, which can relieve symptoms.
  • the other therapeutic agent is an antibody or antigen-binding fragment or an antibody drug conjugate (ADC).
  • ADC antibody drug conjugate
  • an anti-OX40 antibody or antigen-binding fragment (or formulation), another therapeutic agent (or formulation) is used in combination with other therapeutic methods to treat tumors or cancer, such as chemotherapy, radiotherapy, and surgical treatment, among others.
  • the anti-OX40 antibody or antigen-binding fragment at least comprises HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, SEQ ID NO One or more of LCDR1 shown in :4, LCDR2 shown in SEQ ID NO:5, and LCDR3 shown in SEQ ID NO:6.
  • the anti-OX40 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, SEQ ID NO: LCDR1 shown in 4, LCDR2 shown in SEQ ID NO:5, and LCDR3 shown in SEQ ID NO:6.
  • the heavy chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:7, which is at least 80% identical to the sequence set forth in SEQ ID NO:7 sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:7; and/or
  • the light chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 8, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 8, or a sequence with SEQ ID NO: 8 Compared with the sequence shown in NO:8, there are one or more amino acid sequences with conservative amino acid substitutions.
  • the heavy chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 7
  • the light chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises SEQ ID The sequence shown in NO:8.
  • the heavy chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:9, a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO:9, or An amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 9; and/or
  • the light chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 10, a sequence having at least 80% identity to the sequence shown in SEQ ID NO: 10, or a sequence shown in SEQ ID NO: 10
  • the sequences are compared to amino acid sequences having one or more conservative amino acid substitutions.
  • the anti-OX40 antibody is antibody M or M-KF
  • the heavy chain of antibody M or M-KF comprises the sequence shown in SEQ ID NO: 9
  • the light chain of antibody M or M-KF Comprising the sequence shown in SEQ ID NO: 10
  • antibody M or M-KF respectively contains two heavy chains with the same sequence and two light chains with the same sequence.
  • Antibody proteins can be expressed in CHO cells or 293 cells through genetic engineering, and obtained by purification; purification can be performed by conventional methods, such as centrifuging the cell suspension first and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as ProteinA affinity column and ion exchange column can be used to further purify antibody protein.
  • the fucosylation level of the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment is 0-10%. In some embodiments, the fucosylation level of the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment is 0-5%.
  • the fucosylation level of the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment is about 0, about 0.1%, about 0.5%, about 0.8%, about 1%, about 1.3%, about 1.6%, about 2.1%, 2.9%, about 3%, about 3.3%, 3.8%, about 4%, about 4.2%, 4.3%, about 4.6%, about 5%, or any two of these values range between values (inclusive) or any value therein.
  • the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment does not bind fucose.
  • the anti-OX40 antibody (such as antibody M-KF) or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
  • hypofucosylated or afucosylated anti-OX40 antibody or antigen-binding fragment is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line.
  • antibody M-KF is expressed by ⁇ -(1,6)-fucosyltransferase knockout CHO cells.
  • the present invention discloses a method for treating a tumor or cancer in a patient in need thereof, comprising administering an effective amount of an anti-OX40 antibody and another therapeutic agent, wherein the effective amount of the anti-OX40 antibody administered is about 0.6 mg to 900 mg per cycle of treatment.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the anti-OX40 antibody is antibody M or M-KF.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • another therapeutic agent or a combination of an anti-OX40 antibody and another therapeutic agent
  • a pharmaceutical composition respectively, and formulated in a suitable
  • Various forms of the chosen route of administration are administered to the patient, for example by parenteral, intravenous (iv), intramuscular, topical or subcutaneous routes.
  • the anti-OX40 antibody and another therapeutic agent can be infused intravenously separately.
  • anti-OX40 antibody and another therapeutic agent will depend on the nature of the drug, the extent to which the cell surface triggers internalization, transport and release of the drug, the disease being treated, and the condition of the patient (eg, age, sex, weight, etc.).
  • the anti-OX40 antibody (eg, Antibody M or M-KF) is about 0.01 mg/kg to 25 mg/kg per administration or formulations containing such doses of anti-OX40 antibody.
  • Preparations containing anti-OX40 antibodies may be those suitable for injectable use including sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
  • suitable carriers include solvents or dispersion media of physiological saline, bacteriostatic water, or phosphate buffered saline (PBS), ethanol, polyalcohols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), and suitable mixture.
  • PBS phosphate buffered saline
  • the formulation comprises at least 0.1% anti-OX40 antibody.
  • the percentage of antibody can vary and can range from about 2% to 90% by weight of a given dosage form.
  • the anti-OX40 antibody per administration is about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg/kg, about 0.06 mg/kg, about 0.08 mg/kg, about 0.1 mg/kg, about 0.2mg/kg, about 0.3mg/kg, about 0.5mg/kg, about 0.9mg/kg, about 1mg/kg, about 2mg/kg, about 2.5mg/kg, about 3mg/kg, about 4mg/kg, about 5mg/kg, about 6mg/kg, about 7mg/kg, about 8mg/kg, about 9mg/kg, about 10mg/kg, about 11mg/kg, about 12mg/kg, about 13mg/kg, about 14mg/kg, about 15mg/kg, about 16mg/kg, about 17mg/kg, about 18mg/kg, about 19mg/kg, about 20mg/kg, about 21mg/kg, about 22mg/kg, about 23mg/kg, about
  • an anti-OX40 antibody (eg, antibody M or M-KF) is administered at an effective dose of about 0.6 mg to 900 mg per dose. In some embodiments, the anti-OX40 antibody is administered at an effective dose of about 0.6 mg to 600 mg per dose. In some embodiments, the effective amount of anti-OX40 antibody (eg, antibody M or M-KF) administered is 0.6 mg to 900 mg once every 2, 3 or 4 weeks. In some embodiments, an effective amount of an anti-OX40 antibody administered is about 0.6 mg, about 1.8 mg, about 6 mg, about 18 mg, about 60 mg, about 100 mg, about 180 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, or about 600mg every 2, 3 or 4 weeks.
  • the effective amount of anti-OX40 antibody administered is about 0.6 mg, about 1.8 mg, about 6 mg, about 18 mg, about 60 mg, about 180 mg, about 360 mg, or about 600 mg once every 2, 3, or 4 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 600 mg to 900 mg once every 2, 3 or 4 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 700 mg, about 750 mg, about 800 mg, about 830 mg, or about 900 mg once every 2, 3, or 4 weeks.
  • therapeutically effective amounts of the other therapeutic agent and the anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the other therapeutic agent and the anti-OX40 antibody may be administered for the same or different periods.
  • the anti-OX40 antibody and another therapeutic agent are administered by subcutaneous (s.c.) injection, intraperitoneal (i.p.) injection, parenteral injection, intraarterial injection, or intravenous (i.v.) injection.
  • the anti-OX40 antibody and the other therapeutic agent can be administered by the same or different means.
  • the anti-OX40 antibody is administered by intravenous (i.v.) infusion or bolus injection.
  • the second therapeutic agent is administered by intravenous (i.v.) infusion or bolus injection.
  • the anti-OX40 antibody (such as antibody M or M-KF) and the other therapeutic agent are independent dosage units and used in combination. In some embodiments, the anti-OX40 antibody can be administered before, after, or simultaneously with the other therapeutic agent. .
  • the anti-OX40 antibody (such as antibody M or M-KF) and the other therapeutic agent simultaneously form a combined administration unit and are administered in combination.
  • the patient has a tumor or cancer.
  • tumors and cancers include, but are not limited to, hematological cancers, solid tumors.
  • hematological cancers include, but are not limited to, leukemias, lymphomas, and myelomas.
  • leukemia includes acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and myeloproliferative disorders/neoplastics (MPDS) ).
  • ALL acute lymphoblastic leukemia
  • AML acute myelogenous leukemia
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • MPDS myeloproliferative disorders/neoplastics
  • lymphoma includes Hodgkin's lymphoma, indolent and aggressive non-Hodgkin's lymphoma, Burkitt's lymphoma, and follicular lymphoma (small and large cell).
  • the myeloma includes multiple myeloma (MM), giant cell myeloma, heavy chain myeloma, and light chain or Bens-Jones myeloma.
  • solid tumors include breast cancer, ovarian cancer, lung cancer, pancreatic cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, head and neck cancer, bladder cancer, esophageal cancer, liver cancer, and kidney cancer.
  • the tumors and cancers are pathologically confirmed locally advanced or metastatic malignant solid tumors for which there is no effective treatment.
  • the other therapeutic agent is selected from the following antibody or antigen-binding fragment or antibody drug conjugate (ADC) against the target: EGFR (epidermal growth factor receptor), VEGF (vascular endothelial growth factor) , VEGFR2 (vascular endothelial growth factor receptor 2), CTLA-4 (cytotoxic T lymphocyte-associated protein 4), PD-1 (programmed death receptor-1), PD-L1 (programmed death ligand-1 ), HER2 (human epidermal growth factor receptor 2), CD20 (cluster of differentiation 20), TROP2 (human trophoblast surface antigen 2), LAG3 (lymphocyte activation gene-3 molecule), CD27 (cluster of differentiation 27), ICOS (inducible costimulator), BTLA (B and T lymphocyte weakening factor), TIM3 (T cell immunoglobulin mucin 3), BCMA (B cell maturation antigen), c-MET (mesenchymal transition factor), TAA (tumor related antigen), GITR (ADC) against
  • the another therapeutic agent is selected from the group consisting of anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-GITR antibodies, anti-TIGIT antibodies, anti-CD19 antibodies, and anti- 4-1BB antibody.
  • the other therapeutic agent is an antibody (anti-CTLA-4 antibody) or antigen-binding fragment targeting T-lymphocyte-associated antigen 4 (CTLA-4), such as ipilimumab (Yervoy TM or its biological analogs) or defucosylated ipilimumab, as described in WO2014089113.
  • CTLA-4 T-lymphocyte-associated antigen 4
  • the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO:23
  • the light chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO:24
  • the anti-CTLA -4 antibodies contain two sequence-identical heavy chains and two sequence-identical light chains.
  • the anti-CTLA-4 antibody or antigen-binding fragment is expressed by CHO cells. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is expressed by an ⁇ -(1,6)-fucosyltransferase knockout cell line (eg, CHO cells).
  • the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-10%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%. In some embodiments, the fucosylation level of the anti-CTLA-4 antibody or antigen-binding fragment is about 0, about 0.1%, about 0.3%, about 0.4%, about 0.6%, about 1.3%, about 1.9 %, about 2.2%, about 2.8%, about 3.3%, about 3.7%, about 4.1%, about 4.5%, about 5%, or a range between any two of these values (inclusive) or any value therein .
  • the anti-CTLA-4 antibody or antigen-binding fragment has ⁇ 5% total high mannose glycoforms and/or ⁇ 3% total sialylated glycoforms.
  • the total amount of high mannose glycoforms of the anti-CTLA-4 antibody or antigen-binding fragment is about 0.1%, about 0.3%, about 0.9%, about 1.18%, about 1.7%, about 2.6%, about 3.3% %, about 4.1%, about 4.9%, about 4.99%, or a range between any two of these values (inclusive), or any value therein.
  • the total amount of sialylated glycoforms of the anti-CTLA-4 antibody or antigen-binding fragment is about 0.1%, about 0.2%, about 0.36%, about 0.8%, about 1.5%, about 2.2%, about 2.7%, about 2.9%, about 2.99%, or a range between any two of these values, inclusive, or any value therein.
  • the anti-CTLA-4 antibody or antigen-binding fragment has ⁇ 2% total high mannose glycoforms and/or ⁇ 1% total sialylated glycoforms.
  • the anti-CTLA-4 antibody is ipilimumab and the effective amount administered is about 30 mg to 300 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of ipilimumab administered is about 1 mg/kg, about 3 mg/kg, or about 10 mg/kg once every 3 weeks, 6 weeks, or 12 weeks.
  • the effective amount of ipilimumab administered is about 1 mg/kg every 3 or 6 weeks.
  • the effective amount of ipilimumab administered is about 3 mg/kg every 3 or 6 weeks.
  • the effective amount of ipilimumab administered is about 10 mg/kg every 3 weeks or every 12 weeks.
  • the amount of ipilimumab per administration is about 0.5 mg/kg to 10 mg/kg or formulations containing such doses of ipilimumab.
  • the ipilimumab per administration is about 0.5 mg/kg, about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 6.9mg/kg, about 7mg/kg, about 8.4mg/kg, about 9mg /kg, about 10 mg/kg, or a range between any two of these values (inclusive), or any value therein, or a formulation containing this dose of ipilimumab.
  • therapeutically effective amounts of ipilimumab and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration cycles of ipilimumab and anti-OX40 antibody can be the same or different.
  • the other therapeutic agent is an anti-PD-1 antibody or antigen-binding fragment.
  • the anti-PD-1 antibody is nivolumab, pembrolizumab, camrelizumab, sintilimab , toripalimab or tislelizumab, or an anti-PD-1 antibody or antigen-binding fragment described in WO2020207432.
  • Anti-PD-1 antibodies or antigen-binding fragments can be expressed in cells (such as CHO) by genetic engineering and obtained by purification.
  • the anti-PD-1 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:25, HCDR2 shown in SEQ ID NO:26, HCDR3 shown in SEQ ID NO:27, SEQ ID LCDR1 shown in NO:28, LCDR2 shown in SEQ ID NO:29 and LCDR3 shown in SEQ ID NO:30.
  • the heavy chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:31, which is at least 80% identical to the sequence set forth in SEQ ID NO:31 or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:31; and/or the light chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises SEQ ID NO:31
  • the sequence shown in ID NO:32 a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO:32, or having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:32 amino acid sequence.
  • the heavy chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:33, and the light chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:34.
  • the effective amount of anti-PD-1 antibody administered is about 50 mg to 600 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of anti-PD-1 antibody administered is about 1 mg/kg to 10 mg/kg every 2 weeks or every 3 weeks.
  • the effective amount of anti-PD-1 antibody administered is 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 (or a range between any two of these values (inclusive) or any value therein) every 2 weeks or every 3 weeks.
  • the effective amount of anti-PD-1 antibody administered is about 100 mg to about 600 mg once every 2, 3 or 4 weeks.
  • the effective amount of the anti-PD-1 antibody administered is about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg (or the range between any two of these values (inclusive). ) or any value therein) every 2, 3, or 4 weeks.
  • the anti-PD-1 antibody is about 1 mg/kg to 10 mg/kg per administration, or a formulation containing such a dose of anti-PD-1 antibody.
  • the anti-PD-1 antibody per administration is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, about 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 7 mg/kg, about 10 mg/kg, or a range (including endpoints) between any two of these values or any value, or preparations containing this dose of anti-PD-1 antibody.
  • therapeutically effective amounts of anti-PD-1 antibody and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration cycles of the anti-PD-1 antibody and the anti-OX40 antibody can be the same or different.
  • the other therapeutic agent is an antibody targeting PD-L1 (anti-PD-L1 antibody) or an antigen-binding fragment, such as Atezolizumab, or Durvalumab (Durvalumab).
  • Anti-PD-L1 antibodies or antigen-binding fragments can be expressed in cells (such as CHO) through genetic engineering, and obtained by purification; purification can be performed by conventional methods.
  • the other therapeutic agent is atezolizumab, including Tecentriq TM , a biosimilar thereof, or an ADCC effect enhancing mAb or an afucosylated mAb.
  • the effective amount of atezolizumab administered is about 60 mg to 1200 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of atezolizumab administered is about 60 mg, 70 mg, 80 mg, 120 mg, 160 mg, 220 mg, 300 mg, 420 mg, 600 mg, 700 mg, 800 mg, 1000 mg, 1120 mg, about 1200 mg once every 3 weeks .
  • each administration of atezolizumab is about 1 mg/kg to 20 mg/kg or a formulation containing such doses of atezolizumab.
  • atezolizumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 9mg/kg, about 12mg/kg, about 15mg/kg, about 18mg/kg, about 20mg/kg , or the range (inclusive) between any two of these values, or any value therein, or a formulation containing this dose of atezolizumab.
  • the other therapeutic agent is Durvalumab, including IMFINZI TM , biosimilars thereof, or ADCC effect enhancing mAbs or afucosylated mAbs.
  • the effective amount of Durvalumab administered is about 60 mg to 900 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of Durvalumab administered is about 5 mg/kg to 15 mg/kg every 2 weeks or every 3 weeks.
  • the effective amount of Durvalumab administered is about 10 mg/kg every 2 weeks.
  • the amount of Durvalumab per administration is from about 1 mg/kg to 10 mg/kg or formulations containing such doses of Durvalumab.
  • each administration of Durvalumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, About 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 6 mg/kg, about 9 mg/kg, about 10 mg/kg, or a range between any two of these values (inclusive) or where Any value, or formulations containing this dose of Durvalumab.
  • therapeutically effective amounts of the anti-PD-L1 antibody and the anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration cycles of the anti-PD-L1 antibody and the anti-OX40 antibody can be the same or different.
  • the other therapeutic agent is an anti-TIGIT antibody or antigen-binding fragment.
  • the anti-TIGIT antibody is Tiragolumab (tireliumab) or its biosimilar or ADCC effect-enhancing monoclonal antibody or defucosylated monoclonal antibody, or the anti-TIGIT antibody described in WO2021043206 or antigen-binding fragments.
  • the anti-TIGIT antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:13, HCDR2 shown in SEQ ID NO:14, HCDR3 shown in SEQ ID NO:15, SEQ ID NO: LCDR1 shown in 16, LCDR2 shown in SEQ ID NO:17, and LCDR3 shown in SEQ ID NO:18.
  • the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO: 19, which is at least 80% identical to the sequence set forth in SEQ ID NO: 19 sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 19; and/or the light chain variable region of the anti-TIGTI antibody or antigen-binding fragment comprises SEQ ID NO: 20
  • the sequence shown is a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO:20, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:20.
  • the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF
  • the heavy chain of antibody h10D8OF and h10D8OFKF comprises the sequence shown in SEQ ID NO: 21
  • the light chain of antibody h10D8OF and h10D8OFKF comprises the sequence shown in SEQ ID NO: 22
  • the sequences are shown; antibodies h10D8OF and h10D8OFKF contain two identical heavy chains and two identical light chains, respectively.
  • the level of fucosylation of the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment is 0-10%. In some embodiments, the level of fucosylation of the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment is 0-5%.
  • the fucosylation level of the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment is about 0, about 0.1%, about 0.5%, about 0.8%, about 1%, about 1.3% , about 1.6%, about 2.1%, 2.9%, about 3%, about 3.3%, 3.8%, about 4%, about 4.2%, about 4.3%, about 4.6%, about 5%, or any two of these values A range between values (inclusive) or any value in it.
  • the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment does not bind fucose.
  • the anti-TIGIT antibody (such as antibody h10D8OFKF) or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
  • the anti-TIGIT antibody or antigen-binding fragment can be expressed in CHO cells or 293 cells by genetic engineering, and obtained by purification; purification can be performed by conventional methods, such as centrifuging the cell suspension first and collecting the supernatant, Centrifuge again to further remove impurities. Methods such as ProteinA affinity column and ion exchange column can be used to purify antibody protein.
  • hypofucosylated or afucosylated anti-TIGIT antibody or antigen-binding fragment is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line.
  • antibody h10D8OFKF is expressed by ⁇ -(1,6)-fucosyltransferase knockout CHO cells.
  • the effective amount of anti-TIGIT antibody administered is about 9 mg to 1200 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of anti-TIGIT antibody administered is about 0.01 mg/kg to 20 mg/kg once every 2 weeks or every 3 weeks.
  • the effective amount of anti-TIGIT antibody administered is about 0.01 mg/kg, about 0.03 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, about 5 mg/kg , about 6mg/kg, about 7mg/kg, about 8mg/kg, about 9mg/kg, about 10mg/kg, about 11mg/kg, about 12mg/kg, about 13mg/kg, about 14mg/kg, about 15mg/kg , about 16 mg/kg, about 17 mg/kg, about 18 mg/kg, about 19 mg/kg, about 20 mg/kg (or the range (including endpoints) between any two of these values or any value therein) per 2 Weekly or every 3 weeks.
  • an effective amount of an anti-TIGIT antibody administered is about 10 mg to about 900 mg once every 2, 3, or 4 weeks. In some embodiments, an effective amount of an anti-TIGIT antibody administered is about 10 mg, about 30 mg, about 100 mg, about 160 mg, about 200 mg, about 242 mg, about 300 mg, about 346 mg, about 400 mg, about 500 mg, about 600 mg (or these values The range between any two values in (inclusive) or any value therein) every 2, 3, or 4 weeks.
  • therapeutically effective amounts of anti-TIGIT antibody and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration periods of the anti-TIGIT antibody and the anti-OX40 antibody may be the same or different.
  • the other therapeutic agent is an anti-GITR antibody or antigen-binding fragment.
  • the anti-GITR antibody or antigen-binding fragment such as the antibody or antigen-binding fragment described in CN111918878A.
  • the effective amount of anti-GITR antibody administered is about 0.01 mg/kg to 20 mg/kg every 2 weeks, every 3 weeks, or every 4 weeks.
  • therapeutically effective amounts of anti-GITR antibody and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration cycles of the anti-GITR antibody and the anti-OX40 antibody may be the same or different.
  • the other therapeutic agent is a monoclonal antibody that specifically binds to the extracellular dimerization domain (subdomain II) of epidermal growth factor receptor 2 (HER2) (anti-HER2 antibody) Or antigen-binding fragments such as Pertuzumab.
  • Pertuzumab can be expressed in cells (such as CHO) by genetic engineering and obtained by purification.
  • the other therapeutic agent is Pertuzumab, including Perjeta TM or a biosimilar thereof or an ADCC effect enhancing mAb or an afucosylated mAb.
  • the effective amount of pertuzumab administered is about 40 mg to 900 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of Pertuzumab administered is about an initial 840 mg followed by 420 mg every 3 weeks thereafter.
  • the amount of Pertuzumab per administration is about 1 mg/kg to 12 mg/kg or a formulation containing such doses of Pertuzumab.
  • pertuzumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, about 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 6 mg/kg, about 6.9 mg/kg, about 7 mg/kg, about 8.4 mg/kg, about 9 mg/kg, about 11 mg/kg, about 12 mg/kg, or a range between any two of these values (inclusive), or any value therein, or a formulation containing such a dose of Pertuzumab.
  • therapeutically effective amounts of pertuzumab and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration periods of Pertuzumab and anti-OX40 antibody can be the same or different.
  • the other therapeutic agent is a recombinant humanized immunoglobulin G1 (IgG1) monoclonal antibody that binds VEGF-A and inhibits its binding to VEGF receptor-2 (VEGFR2) (anti- VEGF antibody), such as bevacizumab.
  • IgG1 humanized immunoglobulin G1
  • VEGFR2 VEGF receptor-2
  • Bevacizumab can be expressed in cells (such as CHO) by genetic engineering and obtained by purification.
  • the other therapeutic agent is bevacizumab, bevacizumab comprising or its biosimilars, such as BAT1706, or ADCC effect enhancing mAb or defucosylated mAb.
  • the effective amount of bevacizumab administered is about 50 mg to 400 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of bevacizumab administered is about 5 mg/kg to 15 mg/kg every 2 weeks or every 3 weeks.
  • the effective amount of bevacizumab administered is about 5 mg/kg, about 7.5 mg/kg, about 10 mg/kg, or about 15 mg/kg every 2 weeks or every 3 weeks.
  • the effective amount of bevacizumab administered is about 5 mg/kg every 2 weeks, 10 mg/kg every 2 weeks, 7.5 mg/kg every 3 weeks, 15 mg/kg every 3 weeks 1 time a week.
  • the amount of bevacizumab per administration is about 1 mg/kg to 9 mg/kg or a formulation containing such doses of bevacizumab.
  • each administration of bevacizumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, about 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 6 mg/kg, about 6.9 mg/kg, about 7 mg/kg, about 8.4 mg/kg, about 9 mg/kg, or these
  • therapeutically effective amounts of bevacizumab and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration cycles of bevacizumab and anti-OX40 antibody can be the same or different.
  • the other therapeutic agent is an antibody targeting CD20 (anti-CD20 antibody), such as ofatumumab, obinutuzumab, BAT4306F described in CN109096399A or BAT4406F described in CN109096399A.
  • Anti-CD20 antibody anti-CD20 antibody
  • Ofatumumab and obinutuzumab can be expressed in cells (such as CHO) through genetic engineering and obtained through purification.
  • the other therapeutic agent is ofatumumab, ofatumumab including Arzerra TM or Its biosimilar or ADCC effect enhancing mAb or afucosylated mAb.
  • ofatumumab is administered in an effective amount of about 10 mg to 2000 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount ofatumumab administered is about 20 mg weekly or monthly.
  • the effective amount ofatumumab administered is about 300 mg initially, 1000 mg after 1 week, and 1000 mg every 4 weeks or every 8 weeks thereafter. In some embodiments, the effective amount ofatumumab administered is about 300 mg initially, 2000 mg after 1 week, and 2000 mg every 1 week or every 4 weeks thereafter.
  • each administration of ofatumumab is about 0.5 mg/kg to 18 mg/kg or a formulation containing such a dose of ofatumumab.
  • ofatumumab per administration is about 0.5 mg/kg, about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 6.9mg/kg, about 7mg/kg, about 8.4mg/kg, about 9 mg/kg, about 11 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15 mg/kg, about 17 mg/kg, about 18 mg/kg, or the range between any two of these values (inclusive) or any value therein, or a formulation containing this dose of ofatumumab.
  • therapeutically effective amounts of ofatumumab and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the administration periods of ofatumumab and anti-OX40 antibody can be the same or different.
  • the other therapeutic agent is obinutuzumab, comprising Its biosimilar, or ADCC effect enhancing monoclonal antibody or defucosylated monoclonal antibody.
  • obinutuzumab is administered in an effective amount of about 10 mg to 2000 mg per treatment cycle.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it.
  • the effective amount of obinutuzumab administered is 100 mg on day 1, 900 mg on day 2, 1000 mg on day 8, day 15, and 1000 mg per cycle thereafter.
  • the effective amount of obinutuzumab administered is 1000 mg each on Day 1, Day 8, and Day 15, and 1000 mg per course thereafter. Each course of treatment can be 1 month or 2 months.
  • the amount of obinutuzumab per administration is from about 0.5 mg/kg to 15 mg/kg or a formulation containing such doses of ofatumumab.
  • ofatumumab per administration is about 0.5 mg/kg, about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 6.9mg/kg, about 7mg/kg, about 8.4mg/kg, about 9 mg/kg, about 11 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15 mg/kg, or the range between any two of these values (including the endpoints) or any value therein, or a dose containing Formulations of falimumab.
  • therapeutically effective amounts of obinutuzumab and anti-OX40 antibody are administered to the patient separately or simultaneously.
  • the dosing cycles of obinutuzumab and anti-OX40 antibody can be the same or different.
  • the other therapeutic agent is an anti-HER2 antibody drug conjugate (HER2-ADC) or an anti-Trop2 antibody drug conjugate (Trop2-ADC), such as ado-trastuzumab emtansine (T-DM1), Trastuzumab deruxtecan (DS-8201), the antibody drug conjugate described in CN103333246B and CN109078181A.
  • HER2-ADC anti-HER2 antibody drug conjugate
  • Trop2-ADC anti-Trop2 antibody drug conjugate
  • T-DM1 ado-trastuzumab emtansine
  • DS-8201 Trastuzumab deruxtecan
  • the present invention discloses a method of treating a tumor or cancer comprising administering to a patient in need thereof an effective amount of an anti-OX40 antibody or antigen-binding fragment (or formulation) and another therapeutic agent (or formulation) ;
  • the effective dose of anti-OX40 antibody or antigen-binding fragment is about 0.6 mg to 900 mg in a single administration (or a preparation containing this dose of anti-OX40 antibody).
  • the other therapeutic agent is an anti-PD-1 antibody or antigen-binding fragment.
  • the anti-PD-1 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:25, HCDR2 shown in SEQ ID NO:26, HCDR3 shown in SEQ ID NO:27, SEQ ID LCDR1 shown in NO:28, LCDR2 shown in SEQ ID NO:29 and LCDR3 shown in SEQ ID NO:30.
  • the heavy chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:31, which is at least 80% identical to the sequence set forth in SEQ ID NO:31 or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:31; and/or the light chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises SEQ ID NO:31
  • the sequence shown in ID NO:32 a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO:32, or having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:32 amino acid sequence.
  • the heavy chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:33, and the light chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:34.
  • the effective amount of anti-PD-1 antibody is about 50 mg to 600 mg in a single administration (or a preparation containing such dose of anti-PD-1 antibody). Dosage schedule and mode of administration depend on the benefit-risk assessment of anti-PD-1 antibody (or formulation), anti-OX40 (or formulation) in certain patient populations and general clinical practice guidelines.
  • the anti-OX40 antibody (such as antibody M or M-KF) is administered to the patient in an effective amount of about 0.6 mg to 900 mg of anti-OX40 antibody (or a preparation containing such a dose of anti-OX40 antibody) per treatment cycle,
  • the effective amount of anti-PD-1 antibody administered to the patient in each treatment cycle is about 50 mg to 600 mg (or a preparation containing this dose of anti-PD-1 antibody).
  • the effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) administered to a patient per treatment cycle is about 0.6 mg, about 1.8 mg, about 6 mg, about 9 mg, about 10 mg, about 12 mg, about 18mg, about 20mg, about 30mg, about 50mg, about 60mg, about 80mg, about 120mg, about 180mg, about 200mg, about 250mg, about 290mg, about 300mg, about 330mg, about 360mg, about 380mg, about 400mg, about 434mg, About 480 mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or a range (including endpoints) between any two of these values or any value therein, or containing this dose of anti Preparations of OX40 Antibodies.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • a treatment cycle is administered once from 1 week to 7 weeks.
  • the effective amount of the anti-OX40 antibody administered in each treatment cycle is 0.6 mg to 100 mg, or a preparation containing this dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or a range between any two of these values (inclusive) or any value therein.
  • one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks.
  • the patient is administered an effective amount of anti-OX40 antibody of about 100 mg to about 300 mg in each treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, About 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of anti-OX40 antibody of about 300 mg to about 600 mg in each treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, About 3 weeks, or about 4 weeks.
  • the patient is administered an effective amount of anti-OX40 antibody of about 600 mg to about 900 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, About 3 weeks or about 4 weeks.
  • the effective amount of the anti-OX40 antibody administered to the patient per treatment cycle is about 6 mg, about 18 mg, about 50 mg, about 60 mg, about 100 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 260 mg, about 300 mg, about 360 mg, about 400 mg, about 430 mg, about 460 mg, about 520 mg, about 600 mg, about 900 mg, or any of these values
  • the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 5 mg to 16 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 6 mg is administered 1 Second-rate.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • the patient is administered an effective amount of anti-OX40 antibody of about 6 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) of about 15 mg to 40 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 18 mg is administered 1 Second-rate.
  • an anti-OX40 antibody eg, antibody M or M-KF
  • the patient is administered an effective amount of anti-OX40 antibody of about 18 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 100 mg to 190 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 180 mg is administered 1 Second-rate.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • the patient is administered an effective amount of anti-OX40 antibody of about 180 mg per treatment cycle, or a preparation containing this dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks week or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 167 mg to 250 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 210 mg is administered 1 Second-rate.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • the patient is administered an effective amount of anti-OX40 antibody of about 210 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 267 mg to 354 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; Second-rate.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • the patient is administered an effective amount of anti-OX40 antibody of about 312 mg, or a preparation containing such a dose of anti-OX40 antibody, in each treatment cycle; wherein, a treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 380 mg to 450 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 400 mg is administered 1 Second-rate.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • the patient is administered with an effective amount of anti-OX40 antibody of about 400 mg in each treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 542 mg to 643 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 620 mg is administered 1 Second-rate.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • the patient is administered an effective amount of anti-OX40 antibody of about 620 mg, or a preparation containing such a dose of anti-OX40 antibody, in each treatment cycle; wherein, a treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 827 mg to 940 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 900 mg is administered 1 Second-rate.
  • an anti-OX40 antibody e.g., antibody M or M-KF
  • the patient is administered an effective amount of anti-OX40 antibody of about 900 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
  • the effective amount of an anti-OX40 antibody is about 0.6 mg to 900 mg once every 3 weeks. In some embodiments, the effective amount of an anti-OX40 antibody is about 0.6 mg, about 1.8 mg, about 10 mg, about 30 mg, about 60 mg, about 100 mg, about 180 mg, about 200 mg, about 300 mg, about 360 mg, about 400 mg, about 500 mg , about 600 mg, about 700 mg, about 800 mg, or about 900 mg every 3 weeks. In some embodiments, the effective amount of an anti-OX40 antibody is about 6 mg, about 18 mg, about 60 mg, about 180 mg, about 360 mg, about 600 mg, or about 900 mg once every 3 weeks.
  • the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 50 mg, about 60 mg, about 80 mg, about 120 mg, about 200 mg, about 250 mg, about 290 mg, about 300 mg, about 330 mg, about 380 mg, about 400 mg, about 434 mg, about 480 mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, or a range between any two of these values (including endpoints) or any value therein, or containing this dose of anti-PD -1 Preparation of antibodies.
  • a treatment cycle is administered once from 1 week to 7 weeks.
  • the effective amount of anti-PD-1 antibody administered in each treatment cycle is 100 mg to 200 mg, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or a range (including endpoints) between any two of these values, or any value therein. In some embodiments, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 200 mg to about 300 mg in each treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; wherein, one treatment cycle is about 1 week, About 2 weeks, about 3 weeks, or about 4 weeks.
  • the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 300 mg, or the range between any two of these values (including the endpoints) or any value therein, or a preparation containing this dose of anti-PD-1 antibody; wherein, one treatment cycle is about 1 weeks, about 2 weeks, about 3 weeks, or about 4 weeks.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 45 mg to 80 mg per treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; for example, about 50 mg is administered once.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 50 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 87 mg to 130 mg per treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; for example, about 100 mg is administered once.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 100 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 180 mg to 230 mg per treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; for example, about 200 mg is administered once.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 200 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
  • the effective amount of anti-PD-1 antibody administered to the patient is about 267 mg to 343 mg, or a preparation containing this dose of anti-PD-1 antibody; for example, about 300 mg is administered once in each treatment cycle.
  • the patient is administered with an effective amount of anti-PD-1 antibody of about 300 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 300 mg to 700 mg per treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; for example, about 600 mg is administered once.
  • the patient is administered an effective amount of anti-PD-1 antibody of about 600 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
  • the patient is administered an anti-OX40 antibody and another therapeutic agent (or a combination of an anti-OX40 antibody and another therapeutic agent) once per treatment cycle.
  • the anti-OX40 antibody and another therapeutic agent or a combination of an anti-OX40 antibody and another therapeutic agent
  • the patient can only be dosed 1 or 4 times per treatment cycle.
  • the patient is treated for one treatment cycle. In some embodiments, the patient receives multiple (e.g., 2, 3, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16 or 17) treatment cycles. In some embodiments, the patient is treated until the condition is in remission and no longer requires treatment.
  • the present invention discloses a method for treating tumor or cancer, said method comprising: administering about 0.6 mg to 100 mg, about 100 mg to 300 mg, about 300 mg to 600 mg, or about 600 mg to a patient in need thereof Up to 900 mg, such as about 0.6 mg, about 1.8 mg, about 6 mg, about 10 mg, about 18 mg, about 20 mg, about 30 mg, about 100 mg, about 120 mg, about 180 mg, about 200 mg, about 300 mg, about 360 mg, about 600 mg, or about 900 mg
  • Anti-OX40 antibody (such as antibody M or M-KF), or preparations containing such doses of anti-OX40 antibody; also administer about 50 mg to 100 mg, about 100 mg to 200 mg, about 200 mg to 400 mg, or about 400 mg to 600 mg to patients in need , such as about 100 mg, about 120 mg, about 150 mg, about 200 mg or about 300 mg of anti-PD-1 antibody, or a preparation containing such a dose of anti-PD-1 antibody.
  • the patient is treated with a single dose of the anti-about 200 mg antibody, and a single dose of the anti-PD-1 antibody. In some embodiments, the patient is treated with a single dose of the anti-OX40 antibody and anti-PD-1 antibody composition.
  • the anti-OX40 antibody and the anti-PD-1 antibody are administered every 3 weeks.
  • the patient's symptoms are relieved. In some embodiments, after a single dose of administration, the patient's symptoms are not relieved as expected, and then about 0.6 mg to 900 mg of the anti-OX40 antibody and about 50 mg to 600 mg of the anti-PD-1 antibody are administered to the patient.
  • the anti-OX40 antibody is Antibody M or M-KF; the anti-PD-1 antibody is Antibody A.
  • the present invention discloses a method for treating a tumor or cancer, the method comprising administering 3 mg/kg to 10 mg/kg of the anti-OX40 antibody M-KF and 300mg to 600mg of anti-PD-1 antibody A.
  • the method comprises administering 3 mg/kg, 5 mg/kg, 6 mg/kg, or 10 mg/kg of Antibody M-KF and 300 mg of Antibody A to a patient in need thereof once every 3 weeks.
  • the method comprises administering 3 mg/kg, 5 mg/kg, 6 mg/kg, or 10 mg/kg of anti-OX40 antibody M-KF and 600 mg of anti-PD-1 once every 3 weeks to a patient in need thereof Antibody A.
  • the method comprises administering to the patient an equivalent amount of Antibody M-KF once 3 weeks prior to administering Antibody M-KF and Antibody A to the patient.
  • the anti-OX40 antibody (or formulation), anti-PD-1 antibody (or formulation) is injected subcutaneously (s.c.), intraperitoneally (i.p.), parenterally, intraarterially, or intravenously (i.v. ) administration by way of injection or the like.
  • the anti-OX40 antibody (or preparation) and anti-PD-1 antibody (or preparation) are administered by infusion.
  • the anti-OX40 antibody (or preparation) and anti-PD-1 antibody (or preparation) are administered by bolus injection.
  • the anti-OX40 antibody (or formulation), anti-PD-1 antibody (or formulation) is administered by intravenous (i.v.) infusion.
  • the duration of the intravenous infusion is about 50 minutes, about 55 minutes, about 60 minutes, about 65 minutes, about 70 minutes, about 75 minutes, about 81 minutes, about 87 minutes, about 90 minutes, about 95 minutes minutes, or the range between any two of these values (inclusive), or any value therein.
  • Figure 1 shows that the anti-OX40 antibody M-KF inhibits the growth of tumor; the ordinate indicates the tumor volume.
  • an entity refers to one or more such entities, for example "an antibody” should be understood as one or more antibodies, therefore, the term “a” (or “an” ), “one or more” and “at least one” may be used interchangeably herein.
  • compositions, methods, etc. include the listed elements, such as components or steps, but not exclude others.
  • Consisting essentially of means that the compositions and methods exclude other elements that substantially affect the characteristics of the combination, but do not exclude elements that do not substantially affect the composition or method.
  • Consisting of means excluding elements not specifically recited.
  • polypeptide is intended to encompass the singular as well as the plural “polypeptides” and refers to a molecule composed of amino acid monomers linked linearly by amide bonds (also known as peptide bonds).
  • polypeptide refers to any chain or chains of two or more amino acids, and does not refer to a specific length of the product.
  • the definition of “polypeptide” includes peptide, dipeptide, tripeptide, oligopeptide, "protein”, “amino acid chain” or any other term used to refer to a chain of two or more amino acids, and the term “polypeptide” may Used in place of, or interchangeably with, any of the above terms.
  • polypeptide is also intended to refer to the products of post-expression modifications of the polypeptide, including but not limited to glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, or non-natural Amino acid modifications that occur.
  • a polypeptide may be derived from natural biological sources or produced by recombinant techniques, but it need not be translated from a specified nucleic acid sequence, it may be produced by any means including chemical synthesis.
  • amino acid refers to an organic compound containing both amino and carboxyl groups, such as an ⁇ -amino acid, which can be encoded by a nucleic acid directly or in the form of a precursor.
  • a single amino acid is encoded by a nucleic acid consisting of three nucleotides (so-called codons or base triplets). Each amino acid is encoded by at least one codon. The fact that the same amino acid is encoded by different codons is called “degeneracy of the genetic code”.
  • Amino acids include natural amino acids and unnatural amino acids.
  • Natural amino acids include alanine (three-letter code: ala, one-letter code: A), arginine (arg, R), asparagine (asn, N), aspartic acid (asp, D), cysteine amino acid (cys, C), glutamine (gln, Q), glutamic acid (glu, E), glycine (gly, G), histidine (his, H), isoleucine (ile, I ), leucine (leu, L), lysine (lys, K), methionine (met, M), phenylalanine (phe, F), proline (pro, P), serine (ser, S), threonine (thr, T), tryptophan (trp, W), tyrosine (tyr, Y) and valine (val, V).
  • a “conservative amino acid substitution” refers to the replacement of one amino acid residue with another amino acid residue containing a side chain (R group) of similar chemical properties (eg, charge or hydrophobicity). In general, conservative amino acid substitutions are unlikely to substantially alter the functional properties of a protein.
  • classes of amino acids that contain chemically similar side chains include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, Arginine and histidine; 6) acidic side chains: aspartic acid and glutamic acid.
  • the number of amino acids in the "conservative amino acid substitution of VL, VH" can be about 1, about 2, about 3, about 4, about 5, about 6, about 8, about 9, about 10, About 11, about 13, about 14, about 15 conservative amino acid substitutions, or a range between any two of these values (inclusive), or any value therein.
  • the number of amino acids for "conservative amino acid substitutions in the heavy or light chain” can be about 1, about 2, about 3, about 4, about 5, about 6, about 8, about 9, about 10 about 11, about 13, about 14, about 15, about 18, about 19, about 22, about 24, about 25, about 29, about 31, about 35, About 38, about 41, about 45 conservative amino acid substitutions, or a range between any two of these values (inclusive), or any value therein.
  • encoding when applied to a polynucleotide refers to a polynucleotide which is said to "encode” a polypeptide which, in its native state or when manipulated by methods well known to those skilled in the art, is transcribed and/or Or translation may result in the polypeptide and/or fragments thereof.
  • Antibodies, antigen-binding fragments or derivatives disclosed in the present invention include but are not limited to polyclonal, monoclonal, multispecific, fully human, humanized, primatized, chimeric antibodies, single chain antibodies, epitope binding Fragments (eg Fab, Fab', F(ab') 2 and scFv).
  • polypeptides or polynucleotides refers to polypeptides or polynucleotides, meaning forms of polypeptides or polynucleotides that do not occur in nature, non-limiting examples may be produced by combination of polynucleotides or polynucleotides that do not normally exist or peptide.
  • Homology refers to the sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing the alignable positions in each sequence. When a position in the sequences being compared is occupied by the same base or amino acid, then the molecules are homologous at that position. The degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences.
  • At least 80% identity is about 80% identity, about 81% identity, about 82% identity, about 83% identity, about 85% identity, about 86% identity, about 87% identity, About 88% identity, about 90% identity, about 91% identity, about 92% identity, about 94% identity, about 95% identity, about 98% identity, about 99% identity, or these A range (inclusive) between any two values in Numeric or any value therein.
  • a polynucleotide or polynucleotide sequence has a certain percentage (eg, 90%, 95%, 98%, or 99%) of "identity” or “sequence identity” with another sequence.
  • “Sex” means that when the sequences are aligned, the percentage of bases (or amino acids) in the two sequences being compared are identical. This alignment percent identity or sequence identity can be determined using visual inspection or software programs known in the art, such as those described by Ausubel et al.eds. (2007) in Current Protocols in Molecular Biology. It is preferred to use the default parameters for the alignment.
  • Biologically equivalent polynucleotides are polynucleotides that share the above indicated percentages of identity and encode a polypeptide having the same or similar biological activity.
  • Antibody and antigen-binding fragment refer to a polypeptide or polypeptide complex that specifically recognizes and binds to an antigen.
  • Antibodies can be whole antibodies and any antigen-binding fragments thereof or single chains thereof.
  • the term “antibody” thus includes any protein or peptide whose molecule contains at least a portion of an immunoglobulin molecule that has the biological activity to bind an antigen.
  • Antibodies and antigen-binding fragments include, but are not limited to, complementarity determining regions (CDRs), heavy chain variable regions (VH), light chain variable regions (VL), heavy chain constant regions of heavy or light chains or ligand-binding portions thereof (CH), light chain constant region (CL), framework region (FR) or any portion thereof, or at least a portion of a binding protein.
  • CDR regions include the CDR regions of the light chain (LCDR1-3) and the CDR regions of the heavy chain (HCDR1-3).
  • antibody includes a wide variety of polypeptides that can be distinguished biochemically.
  • classes of heavy chains include gamma, mu, alpha, delta or epsilon ( ⁇ , ⁇ , ⁇ , ⁇ , ⁇ ), with some subclasses (eg ⁇ 1- ⁇ 4).
  • the nature of this chain determines the "class” of the antibody as IgG, IgM, IgA, IgG or IgE, respectively.
  • the immunoglobulin subclasses (isotypes), eg, IgGl, IgG2, IgG3, IgG4, IgG5, etc., are well characterized and the functional specificities conferred are also known. All immunoglobulin classes are within the scope of the present disclosure. In some embodiments, the immunoglobulin molecule is of the IgG class.
  • Light chains can be classified as kappa ( ⁇ ) or lambda ( ⁇ ). Each heavy chain can be associated with a kappa or lambda light chain.
  • ⁇ kappa
  • lambda ⁇
  • Each heavy chain can be associated with a kappa or lambda light chain.
  • immunoglobulins are produced by hybridomas, B cells, or genetically engineered host cells, their light and heavy chains are joined by covalent bonds, and the "tail" portions of the two heavy chains are linked by covalent disulfide bonds or non-covalent bonding.
  • the amino acid sequence extends from the N-terminus at the forked end of the Y configuration to the C-terminus at the bottom of each chain.
  • the variable region of the immunoglobulin kappa light chain is V ⁇ ; the variable region of the immunoglobulin lambda light chain is V ⁇ .
  • variable light and variable heavy chains are divided into regions of structural and functional homology.
  • the terms "constant” and “variable” are used according to function.
  • the variable light (VL) and variable heavy (VH) chains determine antigen recognition and specificity.
  • the constant region (CL) of the light chain and the constant region (CH) of the heavy chain confer important biological properties such as secretion, transplacental movement, Fc receptor binding, complement fixation, etc.
  • the N-terminal portion is the variable region and the C-terminal portion is the constant region; the CH3 and CL domains actually comprise the carboxy-terminal ends of the heavy and light chains, respectively.
  • CDR complementarity determining regions
  • CDRs as defined by Kabat and Chothia include overlapping or subsets of amino acid residues when compared to each other. Nevertheless, it is within the scope of the invention to use either definition to refer to the CDRs of an antibody or variant thereof.
  • the exact residue numbers comprising a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in the art can generally determine which specific residues are included in the CDRs based on the amino acid sequence of the variable region of the antibody.
  • Kabat et al. also defined a numbering system applicable to the variable region sequences of any antibody.
  • One of ordinary skill in the art can apply this "Kabat numbering" system to any variable region sequence independently of other experimental data other than the sequence itself.
  • “Kabat numbering” refers to the numbering system proposed by Kabat et al., U.S. Dept. of Health and Human Services in "Sequence of Proteins of Immunological Interest” (1983).
  • Antibodies can also use the EU or Chothia numbering system.
  • an antibody drug conjugate refers to an antibody or antigen-binding fragment thereof chemically linked to one or more chemical drugs (which may optionally be therapeutic or cytotoxic agents).
  • an ADC includes an antibody, a cytotoxic or therapeutic drug, and a linker that enables linking or conjugation of the drug to the antibody.
  • ADCs typically have 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 drug conjugated to the antibody.
  • Drugs that can be included in ADCs are, but not limited to: mitotic inhibitors, antitumor antibiotics, immunomodulators, vectors for gene therapy, alkylating agents, antiangiogenic agents, antimetabolites, boron-containing agents, chemoprotectants, Hormones, antihormones, corticosteroids, photoactive therapeutics, oligonucleotides, radionuclide agents, topoisomerase inhibitors, tyrosine kinase inhibitors, and radiosensitizers.
  • the drug included in the ADC may be a maytansinoid drug.
  • the drug included in the ADC may be a compound represented by formula I as described in this application or a pharmaceutically acceptable salt thereof.
  • the antibody in an ADC, is conjugated to the drug via a self-cysteine or a thiolated amino acid, such as a thiolated lysine, forming a disulfide bond.
  • Treatment means therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent, slow down, ameliorate and stop undesirable physiological changes or disorders, such as the progression of disease, including but not limited to the following whether detectable or undetectable Relief of symptoms, reduction of disease extent, stabilization of disease state (i.e. not worsening), delay or slowing of disease progression, improvement or palliation of disease state, alleviation or disappearance (whether partial or total), prolongation and Expected survival without treatment, etc.
  • Patients in need of treatment include those who already have a condition or disorder, are susceptible to a condition or disorder, or are in need of prevention of the condition or disorder, and can or are expected to benefit from the administration of an antibody or composition disclosed herein for detection, Patients who benefit from the diagnostic process and/or treatment.
  • Patient refers to any mammal in need of diagnosis, prognosis, or treatment, including humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, and the like. In some embodiments, the patient is a human.
  • Effective amount means that amount of an active compound or agent that elicits a biological or medical response in a tissue, system, animal, individual or human; an effective amount is sought by a researcher, veterinarian, physician or other clinician.
  • the phrase "in need thereof” means that a patient has been identified as being in need of a particular method or treatment. In some embodiments, identification may be by any diagnostic means. A patient may in need thereof during any of the methods and treatments described herein.
  • an antibody expression vector includes at least one promoter element, an antibody coding sequence, a transcription termination signal, and a polyA tail.
  • Other elements include enhancers, Kozak sequences, and donor and acceptor sites for RNA splicing flanking the inserted sequence.
  • High-efficiency transcription can be obtained through the early and late promoters of SV40, long terminal repeats from retroviruses such as RSV, HTLV1, HIVI, and early promoters of cytomegalovirus, and other cellular promoters such as muscle Kinetin promoter.
  • Suitable expression vectors may include pIRES1neo, pRetro-Off, pRetro-On, PLXSN, or pLNCX, pcDNA3.1(+/-), pcDNA/Zeo(+/-), pcDNA3.1/Hygro(+/-), PSVL, PMSG, pRSVcat, pSV2dhfr, pBC12MI and pCS2 etc.
  • Commonly used mammalian cells include 293 cells, Cos1 cells, Cos7 cells, CV1 cells, mouse L cells and CHO cells, etc.
  • DNA sequences of the heavy chain and light chain according to the amino acid sequences of the heavy chain and light chain of the antibody, and modify the 5' and 3' ends of the DNA sequence with PCR primers, which include the kozak sequences of the heavy chain and the light chain and the Signal peptide DNA sequence.
  • the constructed sequence is then cloned into the existing recombinant antibody expression vector, and the correct construction of the recombinant plasmid is verified by sequencing analysis.
  • the above-mentioned recombinant plasmids were transfected into expression cells for expression, and the supernatant was collected and purified to obtain antibody protein samples, which were used in various examples below.
  • the expression vector used is pCDNA3.1TM(+) (Invitrogen Company, product number is V79020), and the expression cell used for antibody M-KF is ⁇ -(1,6)-fucus Glycosyltransferase gene knockout CHO cells (fucosylation level is about 0), the expression cells used for antibody M are CHO-S cells (purchased from Invitrogen); positive control antibody 11D4 (anti-OX40 antibody) heavy
  • the sequence of chain and light chain comes from patent US8236930B2
  • the expression vector used is pCHO1.0 plasmid (purchased from Invitrogen), and the expression cell is CHO-S cell (purchased from Invitrogen); the amino acid sequences of antibodies M and M-KF are shown in Table 1 , the nucleic acid sequences of antibodies M and M-KF are shown in Table 2, and the sequence of the leader peptide is in the box in Table 2;
  • the expression vector used was pCDNA3.1 TM (+)
  • the expression cell used for the antibody h10D8OF was CHO cells
  • the expression cell used for the antibody h10D8OFKF was ⁇ -(1,6)-fucose CHO cells with base transferase gene knockout (fucosylation level is about 0);
  • the amino acid sequences of antibodies h10D8OF and h10D8OFKF are shown in Table 3;
  • the expression vector used was pCDNA3.1 TM (+), and the expression cell was ⁇ -(1,6)-fucosyltransferase gene knockout CHO cells (Fucus The glycosylation level is about 0); the amino acid sequence of the anti-CTLA-4 antibody is shown in Table 4;
  • the expression vector used for antibody A is pCDNA3.1 TM (+), and the expression cell is CHO cells; the amino acid sequence of antibody A is shown in Table 5.
  • Embodiment 2 drug efficacy test in vivo
  • the anti-tumor efficacy of the antibody M-KF of the present invention was studied in the OX40 humanized mouse model (purchased from Biocytogen). MC38 tumor cells were inoculated subcutaneously on the right side of OX40 humanized female mice at a concentration of 5 ⁇ 10 5 cells/0.1 mL.
  • the tumor grew to 119mm 3 they were randomly divided into groups according to the tumor volume, with 6 rats in each group.
  • the grouping day was defined as D0 day, and the administration began on D0 day; the administration dates were: day 0, day 3, day 6, day 9, day 12, and day 15.
  • the administration route of the antibody was intraperitoneal administration (ip), according to the frequency of Q3D (once every 3 days), three dosage groups of 1 mg/kg, 0.2 mg/kg, and 0.04 mg/kg were administered 6 times in total.
  • the dosing regimen is shown in Table 6.
  • tumors and body weights were measured twice weekly throughout the study and mice were euthanized when tumors reached endpoint or when mice had 20% body weight loss.
  • the tumor volume (mm 3 ) was 0.5 ⁇ (tumor long diameter ⁇ tumor short diameter 2 ).
  • mice The experimental results such as tumor volume, mouse body weight, and tumor weight of mice in each group are expressed as mean ⁇ standard error (mean ⁇ SEM). Data were analyzed using SPSS. P ⁇ 0.05 means significant difference.
  • TGItv inhibition rate relative to tumor volume
  • TGItv (1-(mean RTV administration group )/(mean RTV control group )) ⁇ 100%; mean RTV administration group : RTV average value of administration group, mean RTV control group : control group RTV average value;
  • RTV n V nt /V n0 ;
  • V nt the tumor volume of the mouse numbered on day t,
  • V n0 the tumor volume of the mouse numbered on day 0
  • RTV n the tumor volume of the mouse numbered The relative volume of the tumor in the mice of n at day t.
  • N is the number of animals.
  • the medium and high doses of antibody M-KF can significantly inhibit the growth of tumors (P ⁇ 0.05), and the effect is significantly better than the high dose of antibody 11D4; Significant differences were found, indicating that the mice tolerated the drug well against antibody M-KF.
  • Example 3 MC38 colon cancer animal model experiment of B-hPD-1/hOX40 double humanized mice
  • MC38 colon cancer cells resuspended in PBS were inoculated into B-hPD-1/hOX40 humanized mice at a concentration of 5 ⁇ 105/0.1mL, 0.1mL/vessel (Beijing Biocytogen Biotechnology Co., Ltd.) Subcutaneous on the right side.
  • the average tumor volume reached about 105 mm 3 72 mice with appropriate individual tumor volumes were selected to enter the group, and the animals were randomly assigned to 9 groups according to the tumor volume, with 8 mice in each group, and the administration began on the day of the grouping.
  • Table 8 For the specific dosage regimen, see Table 8 below.
  • Drug evaluation indicators include: tumor volume inhibition rate (TGItv) and tumor weight inhibition rate (TGItw):
  • TGItv (%) [1-(Ti-T0)/(Vi-V0)] ⁇ 100%
  • Ti the mean value of the tumor volume of the treatment group on the i-th day of administration
  • T0 the treatment group on the 0th day of administration
  • Vi the mean value of the tumor volume of the negative control group on the i-th day of administration
  • V0 the mean value of the tumor volume of the negative control group on the day 0 of the administration
  • TGItw (%) (W negative control group ⁇ W treatment group)/W negative control group ⁇ 100%, W refers to tumor weight.
  • mice had good activity and eating status during the administration period, and their body weight increased to a certain extent, indicating that the mice had good tolerance to the test drug
  • This study is an evaluation of the safety, tolerability, and pharmacokinetics (PK) of antibody M-KF single administration and antibody M-KF combined with anti-PD-1 antibody A in patients with advanced malignant solid tumors.
  • MTD maximum tolerated dose
  • MAD maximum administered dose
  • Antibody M-KF is administered through intravenous infusion, the infusion time is 60 ⁇ 5min, and 21 days after administration is the DLT observation period; the dose of antibody M-KF is 0.1mg/kg, 0.3mg/kg, 1mg/kg kg, 3mg/kg, 6mg/kg and 10mg/kg every 3 weeks. After the first administration of antibody M-KF, antibody A was administered by intravenous infusion at a dose of 300 mg every 3 weeks, and the infusion time was 60 ⁇ 5 minutes. If DLT does not occur in the first cycle (course) of antibody M-KF monotherapy at each dose level, the combination therapy of 300 mg antibody A and antibody M-KF will start from the second cycle.
  • Antibody A was administered at intervals of once every 3 weeks after the injection of antibody M-KF.
  • a single subject will be enrolled at a starting dose level of 0.1 mg/kg antibody M-KF. If there is no grade 2 or higher toxicity that may be related to the study drug during the DLT observation period according to CTCAE v5.0, the dose of the single subject will be increased to 0.3 mg/kg; if a single subject is tested during the DLT observation period If any grade 2 or higher toxicity related to the study drug occurs, an additional 2 subjects will be enrolled at this dose level. In antibody M-KF monotherapy and combination therapy, the observation period of DLT was 3 weeks.
  • Antibody M-KF starts with a dose of 1 mg/kg and adopts a dose-escalation rule based on "3+3" to explore a safe dose range; the dose of antibody M-KF will be increased to 10 mg/kg until reaching the MTD or MAD.
  • the MTD was defined as the highest dose level at which no more than 1 of 6 subjects experienced a dose-limiting toxicity (DLT) during the first cycle of study treatment (21 days after the first dose of antibody M-KF). Dose escalation was based on observed toxicities during the first and second cycles of treatment.
  • DLT dose-limiting toxicity
  • the maximum duration of treatment is 1 year.
  • Subjects who continue to receive treatment for more than 1 year without disease progression can continue to receive the next cycle of drug treatment for more than 2 years. If the patient has an infusion-related reaction and can continue the treatment, diphenhydramine or acetaminophen can be used for prophylaxis based on the actual clinical situation.
  • Safety evaluation indicators involve: vital signs and physical examination, laboratory examination (blood routine, blood biochemistry, thyroid function, coagulation routine, urine routine, stool routine, pregnancy test), electrocardiogram, adverse events (including immune-related adverse events), etc. .
  • PK pharmacokinetic
  • Clinical effectiveness evaluation best overall response (Best overall response), objective response rate (Objective Response Rate, ORR), duration of response (Duration of Response, DOR), disease control rate (Disease control rate, DCR), no progression Survival (Progression-Free Survival, PFS), overall survival (Overall survival, OS).
  • DLT was defined as the toxicity that occurred during the DLT observation period (21-day first cycle of antibody M-KF monotherapy, and 21-day second cycle of antibody A combination therapy). Unless clearly related to the underlying disease, disease progression, concomitant drugs, or comorbidities, all AEs of the assigned grade should be counted as DLTs and considered at least possibly related to the study drug, as follows:
  • Grade 4 neutropenia lasting for any duration, or Grade 3 neutropenia lasting ⁇ 7 days or pre-existing infection

Abstract

Disclosed is an application of an anti-OX40 antibody in a combined drug. The treatment method comprises administering an effective amount of an anti-OX40 antibody and another therapeutic agent to a patient in need thereof.

Description

抗OX40抗体在联合用药中的应用Application of Anti-OX40 Antibody in Combination Drugs 技术领域technical field
本发明属于生物医药领域，尤其涉及抗OX40抗体在联合用药中的应用。The invention belongs to the field of biomedicine, in particular to the application of anti-OX40 antibody in combined medicine.
背景技术Background technique
免疫系统的检查点有两类，一类是抑制性的，如PD-1，一类是激活性的，如OX40。免疫系统中T细胞的充分激活，需要两级信号。第一信号由T细胞抗原受体识别抗原产生，经由CD3分子将激活信号转至细胞内；而第二信号被称为共刺激信号，由抗原呈递细胞或者靶细胞表面的共刺激分子与激活的T细胞表面的共刺激分子受体相互作用而产生。共刺激信号促进抗原特异性T细胞增殖和分化为效应T细胞(Lindsay K等Immunity 2016,44(5):1005-1019)。There are two types of checkpoints in the immune system, one is inhibitory, such as PD-1, and the other is activating, such as OX40. Adequate activation of T cells in the immune system requires two levels of signaling. The first signal is generated by T cell antigen receptor recognition of antigen, and the activation signal is transferred into the cell through the CD3 molecule; while the second signal is called costimulatory signal, which is generated by costimulatory molecules on the surface of antigen-presenting cells or target cells and activated Produced by the interaction of co-stimulatory molecule receptors on the surface of T cells. Costimulatory signals promote the proliferation and differentiation of antigen-specific T cells into effector T cells (Lindsay K et al. Immunity 2016,44(5):1005-1019).
OX40，也称为TNFRSF4、ACT35、CD134等，属于肿瘤坏死因子受体超家族(TNFRSF)，其是一种I型跨膜糖蛋白。OX40不在静息的T细胞上表达，而是表达于被激活的CD4 +T细胞、CD8 +T细胞、NK细胞和NKT细胞上(Paterson DJ等Mol.Immunol.1987；24:1281-1290)。T细胞被抗原激活后，会在1-3天内高表达OX40分子。而OX40信号的激活会进一步增强T细胞的激活信号，以增强免疫系统的反应(Gramaglia I等J.Immunol.2000；165:3043-3050)。 OX40, also known as TNFRSF4, ACT35, CD134, etc., belongs to the tumor necrosis factor receptor superfamily (TNFRSF), which is a type I transmembrane glycoprotein. OX40 is not expressed on resting T cells, but on activated CD4 + T cells, CD8 + T cells, NK cells and NKT cells (Paterson DJ et al. Mol. Immunol. 1987; 24:1281-1290). After T cells are activated by antigens, they will highly express OX40 molecules within 1-3 days. The activation of OX40 signal will further enhance the activation signal of T cells to enhance the response of the immune system (Gramaglia I et al. J. Immunol. 2000; 165:3043-3050).
目前，普遍认为，抗OX40抗体主要通过以下三种细胞生理机制激活免疫系统和抑制肿瘤。一是，通过直接激活CD4 +和CD8 +的效应T细胞，促进它们的增殖和生存，以及分泌相关的炎性因子；二是，通过抑制Treg的信号和活性，从而减弱其对免疫系统的抑制效果；三是，通过ADCC或ADCP等，耗竭Treg细胞，减少其对效应T细胞的抑制(J.Willoughby等Molecular Immunology 83,2017,13-22)。 At present, it is generally believed that anti-OX40 antibodies mainly activate the immune system and inhibit tumors through the following three cellular physiological mechanisms. One is to directly activate CD4 + and CD8 + effector T cells, promote their proliferation and survival, and secrete related inflammatory factors; the other is to weaken the suppression of the immune system by inhibiting the signal and activity of Treg The third is to deplete Treg cells through ADCC or ADCP, etc., to reduce their suppression of effector T cells (J. Willoughby et al. Molecular Immunology 83, 2017, 13-22).
OX40是肿瘤免疫疗法中的一个非常有潜力的激活性靶点，能为肿瘤免疫治疗提供新的手段。OX40 is a very potential activating target in tumor immunotherapy, which can provide a new means for tumor immunotherapy.
发明内容Contents of the invention
本发明公开了抗OX40抗体或抗原结合片段用于联合治疗肿瘤或癌症的方法或用途。在一些实施方案中，所述方法或用途包括：向有需要的患者施用有效量的抗OX40抗体或抗原结合片段和另一个治疗剂。另一方面，本发明公开了抗OX40抗体(例如抗体M或M-KF)或抗原结合片段和另一个治疗剂在制备用于治疗肿瘤或癌症的药物 中的应用。The invention discloses a method or application of anti-OX40 antibody or antigen-binding fragment for combined treatment of tumor or cancer. In some embodiments, the method or use comprises: administering to a patient in need thereof an effective amount of an anti-OX40 antibody or antigen-binding fragment and another therapeutic agent. In another aspect, the present invention discloses the use of an anti-OX40 antibody (such as antibody M or M-KF) or an antigen-binding fragment and another therapeutic agent in the preparation of a medicament for treating tumor or cancer.
另一方面，本发明公开了抗OX40抗体(例如抗体M或抗体M-KF)或抗原结合片段在制备和另一种治疗剂联合使用的药物中的用途。In another aspect, the present invention discloses the use of an anti-OX40 antibody (such as Antibody M or Antibody M-KF) or an antigen-binding fragment for the preparation of a medicament for use in combination with another therapeutic agent.
另一方面，本发明公开了抗OX40抗体(例如抗体M或抗体M-KF)或抗原结合片段和另一种治疗剂联合使用在治疗肿瘤或癌症中的用途。In another aspect, the present invention discloses the use of an anti-OX40 antibody (such as antibody M or antibody M-KF) or an antigen-binding fragment in combination with another therapeutic agent in the treatment of tumor or cancer.
另一方面，本发明还公开了试剂盒，所述试剂盒包含抗OX40抗体或抗原结合片段(或制剂)、另一个治疗剂(或制剂)，以及用于指导有需要患者给药抗OX40抗体或抗原结合片段(或制剂)和另一个治疗剂(或制剂)的说明书。在一些实施方案中，本发明还公开了试剂盒，所述试剂盒包含抗OX40抗体或抗原结合片段和另一个治疗剂的组合物(或制剂)以及用于指导有需要患者给药抗OX40抗体或抗原结合片段和另一个治疗剂的组合物(或制剂)的说明书。On the other hand, the present invention also discloses a kit, which comprises an anti-OX40 antibody or an antigen-binding fragment (or preparation), another therapeutic agent (or preparation), and an anti-OX40 antibody for guiding patients in need thereof. or instructions for an antigen-binding fragment (or formulation) and another therapeutic agent (or formulation). In some embodiments, the present invention also discloses a kit comprising a composition (or preparation) of an anti-OX40 antibody or an antigen-binding fragment and another therapeutic agent and a method for guiding administration of an anti-OX40 antibody to a patient in need thereof. or instructions for a composition (or formulation) of an antigen-binding fragment and another therapeutic agent.
另一方面，本发明还公开了包含抗OX40抗体或抗原结合片段和另一个治疗剂的适合注射用的药物组合物，如推注型药物组合物或输液(滴注)型药物组合物。在一些实施方案中，药物组合物至少包含0.1％的抗OX40抗体或抗原结合片段和0.1％的另一种治疗剂。抗体或抗原结合片段和另一个治疗剂的百分比可以变化，可以为给定剂型重量的约2％和约90％之间。这种治疗上有用的药物组合物中抗OX40抗体或抗原结合片段和另一个治疗剂的量可以为给药的有效量。On the other hand, the present invention also discloses a pharmaceutical composition suitable for injection, such as a bolus injection type pharmaceutical composition or an infusion (drip) type pharmaceutical composition, comprising an anti-OX40 antibody or an antigen-binding fragment and another therapeutic agent. In some embodiments, the pharmaceutical composition comprises at least 0.1% of an anti-OX40 antibody or antigen-binding fragment and 0.1% of another therapeutic agent. The percentage of antibody or antigen-binding fragment and another therapeutic agent can vary and can be between about 2% and about 90% by weight of a given dosage form. The amount of anti-OX40 antibody or antigen-binding fragment and another therapeutic agent in such therapeutically useful pharmaceutical compositions can be an effective amount for administration.
另一方面，本发明还公开了上述药物组合物的制备方法：分别将本文所述的抗OX40抗体或抗原结合片段和另一个治疗剂(或抗OX40抗体或抗原结合片段和另一个治疗剂的组合物)与药学上可接受的适合注射用的载体(例如注射用水，生理盐水等)混合。上述抗OX40抗体或抗原结合片段和另一个治疗剂与药学上可接受的载体的混合方法是本领域通常已知的。On the other hand, the present invention also discloses a preparation method of the above-mentioned pharmaceutical composition: separately combining the anti-OX40 antibody or antigen-binding fragment described herein and another therapeutic agent (or the anti-OX40 antibody or antigen-binding fragment and another therapeutic agent) Composition) mixed with a pharmaceutically acceptable carrier suitable for injection (such as water for injection, physiological saline, etc.). The method of mixing the above-mentioned anti-OX40 antibody or antigen-binding fragment and another therapeutic agent with a pharmaceutically acceptable carrier is generally known in the art.
在一些实施方案中，抗OX40抗体或抗原结合片段联合另一个治疗剂用于治疗肿瘤或癌症。本发明将抗OX40抗体或抗原结合片段(或制剂)和另一个治疗剂(或制剂)用于肿瘤或癌症治疗中，可以减缓症状。In some embodiments, an anti-OX40 antibody or antigen-binding fragment is used in combination with another therapeutic agent to treat a tumor or cancer. In the present invention, anti-OX40 antibody or antigen-binding fragment (or preparation) and another therapeutic agent (or preparation) are used in the treatment of tumor or cancer, which can relieve symptoms.
在一些实施方案中，所述另一个治疗剂为抗体或抗原结合片段或抗体药物偶联物(ADC)。In some embodiments, the other therapeutic agent is an antibody or antigen-binding fragment or an antibody drug conjugate (ADC).
在一些实施方案中，抗OX40抗体或抗原结合片段(或制剂)、另一个治疗剂(或制剂)与其他治疗方法联合用于治疗肿瘤或癌症，例如化疗、放疗和手术治疗等。In some embodiments, an anti-OX40 antibody or antigen-binding fragment (or formulation), another therapeutic agent (or formulation) is used in combination with other therapeutic methods to treat tumors or cancer, such as chemotherapy, radiotherapy, and surgical treatment, among others.
在一些实施方案中，所述抗OX40抗体或抗原结合片段至少包含SEQ ID NO:1所示的HCDR1、SEQ ID NO:2所示的HCDR2、SEQ ID NO:3所示的HCDR3、SEQ ID NO:4 所示的LCDR1、SEQ ID NO:5所示的LCDR2、SEQ ID NO:6所示的LCDR3中一个或多个。In some embodiments, the anti-OX40 antibody or antigen-binding fragment at least comprises HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, SEQ ID NO One or more of LCDR1 shown in :4, LCDR2 shown in SEQ ID NO:5, and LCDR3 shown in SEQ ID NO:6.
在一些实施方案中，所述抗OX40抗体或抗原结合片段包含SEQ ID NO:1所示的HCDR1、SEQ ID NO:2所示的HCDR2、SEQ ID NO:3所示的HCDR3、SEQ ID NO:4所示的LCDR1、SEQ ID NO:5所示的LCDR2和SEQ ID NO:6所示的LCDR3。In some embodiments, the anti-OX40 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, SEQ ID NO: LCDR1 shown in 4, LCDR2 shown in SEQ ID NO:5, and LCDR3 shown in SEQ ID NO:6.
在一些实施方案中，所述抗OX40抗体或抗原结合片段的重链可变区包含SEQ ID NO:7所示的序列，与SEQ ID NO:7所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:7所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或In some embodiments, the heavy chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:7, which is at least 80% identical to the sequence set forth in SEQ ID NO:7 sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:7; and/or
所述抗OX40抗体或抗原结合片段的轻链可变区包含SEQ ID NO:8所示的序列，与SEQ ID NO:8所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:8所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 8, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 8, or a sequence with SEQ ID NO: 8 Compared with the sequence shown in NO:8, there are one or more amino acid sequences with conservative amino acid substitutions.
在一些实施方案中，所述抗OX40抗体或抗原结合片段的重链可变区包含SEQ ID NO:7所示的序列，所述抗OX40抗体或抗原结合片段的轻链可变区包含SEQ ID NO:8所示的序列。In some embodiments, the heavy chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 7, and the light chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises SEQ ID The sequence shown in NO:8.
在一些实施方案中，所述抗OX40抗体或抗原结合片段的重链包含SEQ ID NO:9所示的序列，与SEQ ID NO:9所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:9所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或In some embodiments, the heavy chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:9, a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO:9, or An amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 9; and/or
所述抗OX40抗体或抗原结合片段的轻链包含SEQ ID NO:10所示的序列，与SEQ ID NO:10所示序列具有至少80％同一性的序列，或与SEQ ID NO:10所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 10, a sequence having at least 80% identity to the sequence shown in SEQ ID NO: 10, or a sequence shown in SEQ ID NO: 10 The sequences are compared to amino acid sequences having one or more conservative amino acid substitutions.
在一些实施方案中，所述抗OX40抗体为抗体M或M-KF，抗体M或M-KF的重链包含如SEQ ID NO:9所示序列，所述抗体M或M-KF的轻链包含如SEQ ID NO:10所示序列；抗体M或M-KF分别含有两条序列相同的重链和两条序列相同的轻链。In some embodiments, the anti-OX40 antibody is antibody M or M-KF, the heavy chain of antibody M or M-KF comprises the sequence shown in SEQ ID NO: 9, and the light chain of antibody M or M-KF Comprising the sequence shown in SEQ ID NO: 10; antibody M or M-KF respectively contains two heavy chains with the same sequence and two light chains with the same sequence.
抗体蛋白可以通过基因工程在CHO细胞或293细胞中表达，并通过纯化获得；纯化可以采用常规方法进行，例如先离心细胞悬液并收集上清液，再次离心进一步去除杂质。ProteinA亲和柱和离子交换柱等方法可以用于进一步纯化抗体蛋白。Antibody proteins can be expressed in CHO cells or 293 cells through genetic engineering, and obtained by purification; purification can be performed by conventional methods, such as centrifuging the cell suspension first and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as ProteinA affinity column and ion exchange column can be used to further purify antibody protein.
在一些实施方案中，所述抗OX40抗体(例如抗体M-KF)或抗原结合片段的岩藻糖基化水平为0-10％。在一些实施方案中，所述抗OX40抗体(例如抗体M-KF)或抗原结合片段的岩藻糖基化水平为0-5％。在一些实施方案中，所述抗OX40抗体(例如抗体M-KF)或抗原结合片段的岩藻糖基化水平为约0、约0.1％、约0.5％、约0.8％、约1％、约1.3％、约1.6％、约2.1％、2.9％、约3％、约3.3％、3.8％、约4％、约4.2％、4.3％、约4.6％、约5％，或这些数值中任何两个值之间的范围(包括端点)或其中任 何值。在一些实施方案中，所述抗OX40抗体(例如抗体M-KF)或抗原结合片段没有结合岩藻糖。在一些实施方案中，所述抗OX40抗体(例如抗体M-KF)或抗原结合片段具有增强的ADCC效应(antibody-dependent cell-mediated cytotoxicity)。In some embodiments, the fucosylation level of the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment is 0-10%. In some embodiments, the fucosylation level of the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment is 0-5%. In some embodiments, the fucosylation level of the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment is about 0, about 0.1%, about 0.5%, about 0.8%, about 1%, about 1.3%, about 1.6%, about 2.1%, 2.9%, about 3%, about 3.3%, 3.8%, about 4%, about 4.2%, 4.3%, about 4.6%, about 5%, or any two of these values range between values (inclusive) or any value therein. In some embodiments, the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment does not bind fucose. In some embodiments, the anti-OX40 antibody (such as antibody M-KF) or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
在一些实施方案中，低岩藻糖基化或无岩藻糖基化的抗OX40抗体或抗原结合片段由α-(1,6)-岩藻糖基转移酶基因敲除的细胞系表达。在一些实施方案中，抗体M-KF由α-(1,6)-岩藻糖基转移酶基因敲除的CHO细胞表达。In some embodiments, the hypofucosylated or afucosylated anti-OX40 antibody or antigen-binding fragment is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line. In some embodiments, antibody M-KF is expressed by α-(1,6)-fucosyltransferase knockout CHO cells.
在一些实施方案中，本发明公开了一种用于治疗有需要患者的肿瘤或癌症的方法，其包括施用有效量的抗OX40抗体和另一个治疗剂，其中抗OX40抗体施用的有效量为约0.6mg至900mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，所述抗OX40抗体为抗体M或M-KF。In some embodiments, the present invention discloses a method for treating a tumor or cancer in a patient in need thereof, comprising administering an effective amount of an anti-OX40 antibody and another therapeutic agent, wherein the effective amount of the anti-OX40 antibody administered is about 0.6 mg to 900 mg per cycle of treatment. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the anti-OX40 antibody is antibody M or M-KF.
在一些实施方案中，可以分别将抗OX40抗体(例如抗体M或M-KF)和另一个治疗剂(或抗OX40抗体和另一个治疗剂的组合物)配制成药物组合物，并以适合于所选给药途径的多种形式向患者给药，例如通过肠胃外、静脉内(iv)、肌肉内、局部或皮下途径。在一些实施方案中，可以分别将抗OX40抗体和另一个治疗剂(或抗OX40抗体和另一个治疗剂的组合物)静脉输注。抗OX40抗体和另一个治疗剂的量将取决于药物的性质，细胞表面触发药物的内在化，运输和释放的程度，所治疗的疾病，患者的状况(如年龄，性别，体重等)。In some embodiments, an anti-OX40 antibody (e.g., antibody M or M-KF) and another therapeutic agent (or a combination of an anti-OX40 antibody and another therapeutic agent) can be formulated into a pharmaceutical composition, respectively, and formulated in a suitable Various forms of the chosen route of administration are administered to the patient, for example by parenteral, intravenous (iv), intramuscular, topical or subcutaneous routes. In some embodiments, the anti-OX40 antibody and another therapeutic agent (or a combination of anti-OX40 antibody and another therapeutic agent) can be infused intravenously separately. The amount of anti-OX40 antibody and another therapeutic agent will depend on the nature of the drug, the extent to which the cell surface triggers internalization, transport and release of the drug, the disease being treated, and the condition of the patient (eg, age, sex, weight, etc.).
在一些实施方案中，每次施用的抗OX40抗体(例如抗体M或M-KF)约0.01mg/kg至25mg/kg或含此剂量抗OX40抗体的制剂。含抗OX40抗体的制剂可以为适于注射用途的制剂包括无菌水性溶液(在此是水溶性的)或分散体以及用于即时制备无菌注射液或分散体的无菌粉末。对于静脉内施用，合适的载体包括生理盐水、抑菌水或磷酸盐缓冲盐水(PBS)、乙醇、多元醇(例如，甘油、丙二醇和液体聚乙二醇等)的溶剂或分散介质，及其适宜的混合物。在一些实施方案中，制剂至少包含0.1％的抗OX40抗体。抗体的百分比可以变化，并且为给定剂型重量可以约2％至90％之间。In some embodiments, the anti-OX40 antibody (eg, Antibody M or M-KF) is about 0.01 mg/kg to 25 mg/kg per administration or formulations containing such doses of anti-OX40 antibody. Preparations containing anti-OX40 antibodies may be those suitable for injectable use including sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include solvents or dispersion media of physiological saline, bacteriostatic water, or phosphate buffered saline (PBS), ethanol, polyalcohols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), and suitable mixture. In some embodiments, the formulation comprises at least 0.1% anti-OX40 antibody. The percentage of antibody can vary and can range from about 2% to 90% by weight of a given dosage form.
在一些实施方案中，每次施用的抗OX40抗体为约0.01mg/kg、约0.02mg/kg、约0.03mg/kg、约0.06mg/kg、约0.08mg/kg、约0.1mg/kg、约0.2mg/kg、约0.3mg/kg、约0.5mg/kg、约0.9mg/kg、约1mg/kg、约2mg/kg、约2.5mg/kg、约3mg/kg、约4mg/kg、约5mg/kg、约6mg/kg、约7mg/kg、约8mg/kg、约9mg/kg、约10mg/kg、约11mg/kg、约12mg/kg、约13mg/kg、约14mg/kg、约15mg/kg、约16mg/kg、约17mg/kg、约18mg/kg、约19mg/kg、约20mg/kg、约21mg/kg、约22mg/kg、约23mg/kg、约24mg/kg、约25mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量抗OX40抗体的制剂。In some embodiments, the anti-OX40 antibody per administration is about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg/kg, about 0.06 mg/kg, about 0.08 mg/kg, about 0.1 mg/kg, about 0.2mg/kg, about 0.3mg/kg, about 0.5mg/kg, about 0.9mg/kg, about 1mg/kg, about 2mg/kg, about 2.5mg/kg, about 3mg/kg, about 4mg/kg, about 5mg/kg, about 6mg/kg, about 7mg/kg, about 8mg/kg, about 9mg/kg, about 10mg/kg, about 11mg/kg, about 12mg/kg, about 13mg/kg, about 14mg/kg, about 15mg/kg, about 16mg/kg, about 17mg/kg, about 18mg/kg, about 19mg/kg, about 20mg/kg, about 21mg/kg, about 22mg/kg, about 23mg/kg, about 24mg/kg, About 25 mg/kg, or a range between any two of these values (inclusive), or any value therein, or a formulation containing such a dose of anti-OX40 antibody.
在一些实施方案中，抗OX40抗体(例如抗体M或M-KF)施用的有效剂量为每剂约0.6mg至900mg。在一些实施方案中，抗OX40抗体施用的有效剂量为每剂约0.6mg至600mg。在一些实施方案中，抗OX40抗体(如抗体M或M-KF)施用的有效量为0.6mg至900mg每2、3或4周一次。在一些实施方案中，抗OX40抗体施用的有效量为约0.6mg、约1.8mg、约6mg、约18mg、约60mg、约100mg、约180mg、约200mg、约300mg、约400mg、约500mg或约600mg每2、3或4周一次。在一些实施方案中，抗OX40抗体施用的有效量为约0.6mg、约1.8mg、约6mg、约18mg、约60mg、约180mg、约360mg或约600mg每2、3或4周一次。在一些实施方案中，抗OX40抗体施用的有效量为约600mg至900mg每2、3或4周一次。在一些实施方案中，抗OX40抗体施用的有效量为约700mg、约750mg、约800mg、约830mg、或约900mg每2、3或4周一次。In some embodiments, an anti-OX40 antibody (eg, antibody M or M-KF) is administered at an effective dose of about 0.6 mg to 900 mg per dose. In some embodiments, the anti-OX40 antibody is administered at an effective dose of about 0.6 mg to 600 mg per dose. In some embodiments, the effective amount of anti-OX40 antibody (eg, antibody M or M-KF) administered is 0.6 mg to 900 mg once every 2, 3 or 4 weeks. In some embodiments, an effective amount of an anti-OX40 antibody administered is about 0.6 mg, about 1.8 mg, about 6 mg, about 18 mg, about 60 mg, about 100 mg, about 180 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, or about 600mg every 2, 3 or 4 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 0.6 mg, about 1.8 mg, about 6 mg, about 18 mg, about 60 mg, about 180 mg, about 360 mg, or about 600 mg once every 2, 3, or 4 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 600 mg to 900 mg once every 2, 3 or 4 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 700 mg, about 750 mg, about 800 mg, about 830 mg, or about 900 mg once every 2, 3, or 4 weeks.
在一些实施方案中，采用治疗有效量的另一个治疗剂和抗OX40抗体分别或者同时施加在患者上。另一个治疗剂和抗OX40抗体的给药周期可以相同或者不同。在一些实施方案中，抗OX40抗体和另一个治疗剂是通过皮下(s.c.)注射、腹膜内(i.p.)注射、肠胃外注射、动脉内注射或静脉内(i.v.)注射等方式进行给药。抗OX40抗体和另一个治疗剂可以通过相同或者不同方式进行给药。In some embodiments, therapeutically effective amounts of the other therapeutic agent and the anti-OX40 antibody are administered to the patient separately or simultaneously. The other therapeutic agent and the anti-OX40 antibody may be administered for the same or different periods. In some embodiments, the anti-OX40 antibody and another therapeutic agent are administered by subcutaneous (s.c.) injection, intraperitoneal (i.p.) injection, parenteral injection, intraarterial injection, or intravenous (i.v.) injection. The anti-OX40 antibody and the other therapeutic agent can be administered by the same or different means.
在一些实施方案中，抗OX40抗体是通过静脉内(i.v.)输液或推注方式进行给药。在一些实施方案中，另一个治疗剂是通过静脉内(i.v.)输液或推注方式进行给药。In some embodiments, the anti-OX40 antibody is administered by intravenous (i.v.) infusion or bolus injection. In some embodiments, the second therapeutic agent is administered by intravenous (i.v.) infusion or bolus injection.
在一些实施方案中，所述抗OX40抗体(例如抗体M或M-KF)和所述另一个治疗剂分别为独立的给药单元，联合用药。在一些实施方案中，所述抗OX40抗体可以在施加所述另一个治疗剂之前给药，也可以在施加所述另一个治疗剂之后给药，也可以与所述另一个治疗剂同时给药。In some embodiments, the anti-OX40 antibody (such as antibody M or M-KF) and the other therapeutic agent are independent dosage units and used in combination. In some embodiments, the anti-OX40 antibody can be administered before, after, or simultaneously with the other therapeutic agent. .
在一些实施方案中，所述抗OX40抗体(例如抗体M或M-KF)和所述另一个治疗剂同时形成组合给药单元，联合用药。In some embodiments, the anti-OX40 antibody (such as antibody M or M-KF) and the other therapeutic agent simultaneously form a combined administration unit and are administered in combination.
在一些实施方案中，患者患有肿瘤或癌症。在一些实施方案中，肿瘤和癌症包括但不限于血液癌症、实体瘤。在一些实施方案中，血液癌症包括但不限于白血病、淋巴瘤和骨髓瘤。在一些实施方案中，白血病包括急性淋巴细胞性白血病(ALL)、急性骨髓性白血病(AML)、慢性淋巴细胞性白血病(CLL)、慢性骨髓性白血病(CML)和骨髓性增生疾病/肿瘤(MPDS)。在一些实施方案中，淋巴瘤包括霍奇金淋巴瘤、无痛性和侵袭性非霍奇金淋巴瘤、伯基特淋巴瘤和滤泡性淋巴瘤(小细胞和大细胞)。在一些实施方案中，骨髓瘤包括多发性骨髓瘤(MM)、巨细胞骨髓瘤、重链骨髓瘤和轻链或本斯-琼斯骨髓瘤。在一些实施方案中，实体瘤包括乳腺癌、卵巢癌、肺癌、胰腺癌、前列腺癌、黑素瘤、结直肠癌、肺癌、头颈癌、膀胱癌、食道癌、肝癌和肾癌。在一些实 施方案中，肿瘤和癌症为尚无有效治疗手段的经病理学确诊的局部晚期或转移性恶性实体肿瘤。In some embodiments, the patient has a tumor or cancer. In some embodiments, tumors and cancers include, but are not limited to, hematological cancers, solid tumors. In some embodiments, hematological cancers include, but are not limited to, leukemias, lymphomas, and myelomas. In some embodiments, leukemia includes acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and myeloproliferative disorders/neoplastics (MPDS) ). In some embodiments, lymphoma includes Hodgkin's lymphoma, indolent and aggressive non-Hodgkin's lymphoma, Burkitt's lymphoma, and follicular lymphoma (small and large cell). In some embodiments, the myeloma includes multiple myeloma (MM), giant cell myeloma, heavy chain myeloma, and light chain or Bens-Jones myeloma. In some embodiments, solid tumors include breast cancer, ovarian cancer, lung cancer, pancreatic cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, head and neck cancer, bladder cancer, esophageal cancer, liver cancer, and kidney cancer. In some embodiments, the tumors and cancers are pathologically confirmed locally advanced or metastatic malignant solid tumors for which there is no effective treatment.
在一些实施方案中，所述另一个治疗剂选自以下针对靶点的抗体或抗原结合片段或抗体药物偶联物(ADC)：EGFR(表皮生长因子受体)、VEGF(血管内皮生长因子)、VEGFR2(血管内皮生长因子受体2)、CTLA-4(细胞毒性T淋巴细胞相关蛋白4)、PD-1(程序性死亡受体-1)、PD-L1(程序性死亡配体-1)、HER2(人表皮生长因子受体2)、CD20(分化簇20)、TROP2(人滋养层细胞表面抗原2)、LAG3(淋巴细胞活化基因-3分子)、CD27(分化簇27)、ICOS(inducible costimulator)、BTLA(B和T淋巴细胞弱化因子)、TIM3(T细胞免疫球蛋白黏液素3)、BCMA(B细胞成熟抗原)、c-MET(间质表皮转化因子)、TAA(肿瘤相关抗原)、GITR(glucocorticoid-induced TNFR-related protein)、TIGIT(T cell immunoreceptor with Ig and ITIM domains)、CD19(分化簇19)和4-1BB(CD137)。在一些实施方案中，所述抗体为抑制型抗体或激动型抗体。In some embodiments, the other therapeutic agent is selected from the following antibody or antigen-binding fragment or antibody drug conjugate (ADC) against the target: EGFR (epidermal growth factor receptor), VEGF (vascular endothelial growth factor) , VEGFR2 (vascular endothelial growth factor receptor 2), CTLA-4 (cytotoxic T lymphocyte-associated protein 4), PD-1 (programmed death receptor-1), PD-L1 (programmed death ligand-1 ), HER2 (human epidermal growth factor receptor 2), CD20 (cluster of differentiation 20), TROP2 (human trophoblast surface antigen 2), LAG3 (lymphocyte activation gene-3 molecule), CD27 (cluster of differentiation 27), ICOS (inducible costimulator), BTLA (B and T lymphocyte weakening factor), TIM3 (T cell immunoglobulin mucin 3), BCMA (B cell maturation antigen), c-MET (mesenchymal transition factor), TAA (tumor related antigen), GITR (glucocorticoid-induced TNFR-related protein), TIGIT (T cell immunoreceptor with Ig and ITIM domains), CD19 (cluster of differentiation 19) and 4-1BB (CD137). In some embodiments, the antibody is an inhibitory antibody or an agonistic antibody.
在一些实施方案中，所述另一个治疗剂选自以下组成的组：抗CTLA-4抗体、抗PD-1抗体、抗PD-L1抗体、抗GITR抗体、抗TIGIT抗体、抗CD19抗体和抗4-1BB抗体。In some embodiments, the another therapeutic agent is selected from the group consisting of anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-GITR antibodies, anti-TIGIT antibodies, anti-CD19 antibodies, and anti- 4-1BB antibody.
在一些实施方案中，所述另一个治疗剂为靶向T淋巴细胞相关抗原4(CTLA-4)抗体(抗CTLA-4抗体)或抗原结合片段，如伊匹单抗(Yervoy TM或其生物类似物)或去岩藻糖基化伊匹单抗，如WO2014089113所述。在一些实施方案中，抗CTLA-4抗体的重链包含如SEQ ID NO:23所示的氨基酸序列，抗CTLA-4抗体的轻链包含如SEQ ID NO:24所示的氨基酸序列；抗CTLA-4抗体含有两条序列相同的重链和两条序列相同的轻链。 In some embodiments, the other therapeutic agent is an antibody (anti-CTLA-4 antibody) or antigen-binding fragment targeting T-lymphocyte-associated antigen 4 (CTLA-4), such as ipilimumab (Yervoy ™ or its biological analogs) or defucosylated ipilimumab, as described in WO2014089113. In some embodiments, the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO:23, and the light chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO:24; the anti-CTLA -4 antibodies contain two sequence-identical heavy chains and two sequence-identical light chains.
在一些实施方案中，抗CTLA-4抗体或抗原结合片段由CHO细胞表达。在一些实施方案中，抗CTLA-4抗体或抗原结合片段由α-(1,6)-岩藻糖基转移酶基因敲除的细胞系(如CHO细胞)表达。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is expressed by CHO cells. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is expressed by an α-(1,6)-fucosyltransferase knockout cell line (eg, CHO cells).
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的岩藻糖基化水平为0-10％。在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的岩藻糖基化水平为0-5％。在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的岩藻糖基化水平为约0、约0.1％、约0.3％、约0.4％、约0.6％、约1.3％、约1.9％、约2.2％、约2.8％、约3.3％、约3.7％、约4.1％、约4.5％、约5％，或这些数值中任何两个值之间的范围(包括端点)或其中任何值。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-10%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%. In some embodiments, the fucosylation level of the anti-CTLA-4 antibody or antigen-binding fragment is about 0, about 0.1%, about 0.3%, about 0.4%, about 0.6%, about 1.3%, about 1.9 %, about 2.2%, about 2.8%, about 3.3%, about 3.7%, about 4.1%, about 4.5%, about 5%, or a range between any two of these values (inclusive) or any value therein .
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的高甘露糖糖型总量<5％ 和/或唾液酸化糖型总量<3％。在一些实施方案中，抗CTLA-4抗体或抗原结合片段的高甘露糖糖型总量为约0.1％、约0.3％、约0.9％、约1.18％、约1.7％、约2.6％、约3.3％、约4.1％、约4.9％、约4.99％，或这些数值中任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的唾液酸化糖型总量为约0.1％、约0.2％、约0.36％、约0.8％、约1.5％、约2.2％、约2.7％、约2.9％、约2.99％，或这些数值中任何两个值之间的范围(包括端点)或其中任何值。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has <5% total high mannose glycoforms and/or <3% total sialylated glycoforms. In some embodiments, the total amount of high mannose glycoforms of the anti-CTLA-4 antibody or antigen-binding fragment is about 0.1%, about 0.3%, about 0.9%, about 1.18%, about 1.7%, about 2.6%, about 3.3% %, about 4.1%, about 4.9%, about 4.99%, or a range between any two of these values (inclusive), or any value therein. In some embodiments, the total amount of sialylated glycoforms of the anti-CTLA-4 antibody or antigen-binding fragment is about 0.1%, about 0.2%, about 0.36%, about 0.8%, about 1.5%, about 2.2%, about 2.7%, about 2.9%, about 2.99%, or a range between any two of these values, inclusive, or any value therein.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的高甘露糖糖型总量<2％和/或唾液酸化糖型总量<1％。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has <2% total high mannose glycoforms and/or <1% total sialylated glycoforms.
在一些实施方案中，所述抗CTLA-4抗体为伊匹单抗，施用的有效量为约30mg至300mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，伊匹单抗施用的有效量为约1mg/kg、约3mg/kg或约10mg/kg每3周、6周、或12周1次。在一些实施方案中，伊匹单抗施用的有效量为约1mg/kg每3周或6周1次。在一些实施方案中，伊匹单抗施用的有效量为约3mg/kg每3周或6周1次。在一些实施方案中，伊匹单抗施用的有效量为约10mg/kg每3周或每12周1次。In some embodiments, the anti-CTLA-4 antibody is ipilimumab and the effective amount administered is about 30 mg to 300 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of ipilimumab administered is about 1 mg/kg, about 3 mg/kg, or about 10 mg/kg once every 3 weeks, 6 weeks, or 12 weeks. In some embodiments, the effective amount of ipilimumab administered is about 1 mg/kg every 3 or 6 weeks. In some embodiments, the effective amount of ipilimumab administered is about 3 mg/kg every 3 or 6 weeks. In some embodiments, the effective amount of ipilimumab administered is about 10 mg/kg every 3 weeks or every 12 weeks.
在一些实施方案中，每次施用的伊匹单抗为约0.5mg/kg至10mg/kg或含此剂量伊匹单抗的制剂。在一些实施方案中，每次施用的伊匹单抗为约0.5mg/kg、约1mg/kg，约1.2mg/kg、约2mg/kg、约2.4mg/kg、约3mg/kg、约3.6mg/kg、约4mg/kg、约4.8mg/kg、约5mg/kg、约5.5mg/kg、约6mg/kg、约6.9mg/kg、约7mg/kg、约8.4mg/kg、约9mg/kg、约10mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量伊匹单抗的制剂。In some embodiments, the amount of ipilimumab per administration is about 0.5 mg/kg to 10 mg/kg or formulations containing such doses of ipilimumab. In some embodiments, the ipilimumab per administration is about 0.5 mg/kg, about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 6.9mg/kg, about 7mg/kg, about 8.4mg/kg, about 9mg /kg, about 10 mg/kg, or a range between any two of these values (inclusive), or any value therein, or a formulation containing this dose of ipilimumab.
在一些实施方案中，采用治疗有效量的伊匹单抗和抗OX40抗体分别或者同时施加在患者上。伊匹单抗和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of ipilimumab and anti-OX40 antibody are administered to the patient separately or simultaneously. The administration cycles of ipilimumab and anti-OX40 antibody can be the same or different.
在一些实施方案中，所述另一个治疗剂为抗PD-1抗体或抗原结合片段。在一些实施方案中，所述抗PD-1抗体为纳武利尤单抗(nivolumab)、帕博利珠单抗(pembrolizumab)、卡瑞利珠单抗(camrelizumab)、信迪利单抗(sintilimab)、特瑞普利单抗(toripalimab)或替雷利珠单抗(tislelizumab)、或WO2020207432中描述的抗PD-1抗体或抗原结合片段。抗PD-1抗体或抗原结合片段可以通过基因工程在细胞(如CHO)中表达，并通过纯化获得。In some embodiments, the other therapeutic agent is an anti-PD-1 antibody or antigen-binding fragment. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, camrelizumab, sintilimab , toripalimab or tislelizumab, or an anti-PD-1 antibody or antigen-binding fragment described in WO2020207432. Anti-PD-1 antibodies or antigen-binding fragments can be expressed in cells (such as CHO) by genetic engineering and obtained by purification.
在一些实施方案中，所述抗PD-1抗体或抗原结合片段包含SEQ ID NO:25所示的HCDR1、SEQ ID NO:26所示的HCDR2、SEQ ID NO:27所示的HCDR3、SEQ ID NO:28 所示的LCDR1、SEQ ID NO:29所示的LCDR2和SEQ ID NO:30所示的LCDR3。In some embodiments, the anti-PD-1 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:25, HCDR2 shown in SEQ ID NO:26, HCDR3 shown in SEQ ID NO:27, SEQ ID LCDR1 shown in NO:28, LCDR2 shown in SEQ ID NO:29 and LCDR3 shown in SEQ ID NO:30.
在一些实施方案中，所述抗PD-1抗体或抗原结合片段的重链可变区包含SEQ ID NO:31所示的序列，与SEQ ID NO:31所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:31所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或所述抗PD-1抗体或抗原结合片段的轻链可变区包含SEQ ID NO:32所示的序列，与SEQ ID NO:32所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:32所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。In some embodiments, the heavy chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:31, which is at least 80% identical to the sequence set forth in SEQ ID NO:31 or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:31; and/or the light chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises SEQ ID NO:31 The sequence shown in ID NO:32, a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO:32, or having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:32 amino acid sequence.
在一些实施方案中，抗PD-1抗体的重链包含如SEQ ID NO:33所示的氨基酸序列，抗PD-1抗体的轻链包含如SEQ ID NO:34所示的氨基酸序列。In some embodiments, the heavy chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:33, and the light chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:34.
在一些实施方案中，抗PD-1抗体施用的有效量为约50mg至600mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，抗PD-1抗体施用的有效量为约1mg/kg至10mg/kg每2周或每3周1次。在一些实施方案中，抗PD-1抗体施用的有效量为约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(或这些数值中的任何两个值之间的范围(包括端点)或其中任何值)每2周或每3周1次。在一些实施方案中，抗PD-1抗体施用的有效量为约100mg至约600mg每2、3或4周一次。在一些实施方案中，抗PD-1抗体施用的有效量为约100mg、约200mg、约300mg、约400mg、约500mg、约600mg(或这些数值中的任何两个值之间的范围(包括端点)或其中任何值)每2周、3周或4周一次。In some embodiments, the effective amount of anti-PD-1 antibody administered is about 50 mg to 600 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of anti-PD-1 antibody administered is about 1 mg/kg to 10 mg/kg every 2 weeks or every 3 weeks. In some embodiments, the effective amount of anti-PD-1 antibody administered is 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 (or a range between any two of these values (inclusive) or any value therein) every 2 weeks or every 3 weeks. In some embodiments, the effective amount of anti-PD-1 antibody administered is about 100 mg to about 600 mg once every 2, 3 or 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered is about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg (or the range between any two of these values (inclusive). ) or any value therein) every 2, 3, or 4 weeks.
在一些实施方案中，每次施用的抗PD-1抗体为约1mg/kg至10mg/kg或含此剂量抗PD-1抗体的制剂。在一些实施方案中，每次施用的抗PD-1抗体为约1mg/kg，约1.2mg/kg、约2mg/kg、约2.4mg/kg、约3mg/kg、约3.6mg/kg、约4mg/kg、约4.8mg/kg、约5mg/kg、约5.5mg/kg、约7mg/kg、约10mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量抗PD-1抗体的制剂。In some embodiments, the anti-PD-1 antibody is about 1 mg/kg to 10 mg/kg per administration, or a formulation containing such a dose of anti-PD-1 antibody. In some embodiments, the anti-PD-1 antibody per administration is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, about 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 7 mg/kg, about 10 mg/kg, or a range (including endpoints) between any two of these values or any value, or preparations containing this dose of anti-PD-1 antibody.
在一些实施方案中，采用治疗有效量的抗PD-1抗体和抗OX40抗体分别或者同时施加在患者上。抗PD-1抗体和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of anti-PD-1 antibody and anti-OX40 antibody are administered to the patient separately or simultaneously. The administration cycles of the anti-PD-1 antibody and the anti-OX40 antibody can be the same or different.
在一些实施方案中，所述另一个治疗剂为是靶向PD-L1的抗体(抗PD-L1抗体)或抗原结合片段，如阿特珠单抗(Atezolizumab)，或度伐利尤单抗(Durvalumab)。抗PD-L1抗体或抗原结合片段可以通过基因工程在细胞(如CHO)中表达，并通过纯化获得；纯化可以采用常规方法进行。In some embodiments, the other therapeutic agent is an antibody targeting PD-L1 (anti-PD-L1 antibody) or an antigen-binding fragment, such as Atezolizumab, or Durvalumab (Durvalumab). Anti-PD-L1 antibodies or antigen-binding fragments can be expressed in cells (such as CHO) through genetic engineering, and obtained by purification; purification can be performed by conventional methods.
在一些实施方案中，所述另一个治疗剂为阿特珠单抗，阿特珠单抗包括 Tecentriq TM、其生物类似物或ADCC效应增强单抗或去岩藻糖基化单抗。在一些实施方案中，阿特珠单抗施用的有效量为约60mg至1200mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，阿特珠单抗施用的有效量为约60mg、70mg、80mg、120mg、160mg、220mg、300mg、420mg、600mg、700mg、800mg、1000mg、1120mg、约1200mg每3周1次。 In some embodiments, the other therapeutic agent is atezolizumab, including Tecentriq ™ , a biosimilar thereof, or an ADCC effect enhancing mAb or an afucosylated mAb. In some embodiments, the effective amount of atezolizumab administered is about 60 mg to 1200 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of atezolizumab administered is about 60 mg, 70 mg, 80 mg, 120 mg, 160 mg, 220 mg, 300 mg, 420 mg, 600 mg, 700 mg, 800 mg, 1000 mg, 1120 mg, about 1200 mg once every 3 weeks .
在一些实施方案中，每次施用的阿特珠单抗为约1mg/kg至20mg/kg或含此剂量阿特珠单抗的制剂。在一些实施方案中，每次施用的阿特珠单抗为约1mg/kg、约1.2mg/kg、约2mg/kg、约2.4mg/kg、约3mg/kg、约3.6mg/kg、约4mg/kg、约4.8mg/kg，约5mg/kg、约5.5mg/kg、约6mg/kg、约9mg/kg、约12mg/kg、约15mg/kg、约18mg/kg、约20mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量阿特珠单抗的制剂。In some embodiments, each administration of atezolizumab is about 1 mg/kg to 20 mg/kg or a formulation containing such doses of atezolizumab. In some embodiments, atezolizumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 9mg/kg, about 12mg/kg, about 15mg/kg, about 18mg/kg, about 20mg/kg , or the range (inclusive) between any two of these values, or any value therein, or a formulation containing this dose of atezolizumab.
在一些实施方案中，所述另一个治疗剂为Durvalumab，包括IMFINZI TM、其生物类似物或ADCC效应增强单抗或去岩藻糖基化单抗。在一些实施方案中，Durvalumab施用的有效量为约60mg至900mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，Durvalumab施用的有效量为约5mg/kg到15mg/kg每2周或每3周1次。在一些实施方案中，Durvalumab施用的有效量为约10mg/kg每2周1次。 In some embodiments, the other therapeutic agent is Durvalumab, including IMFINZI ™ , biosimilars thereof, or ADCC effect enhancing mAbs or afucosylated mAbs. In some embodiments, the effective amount of Durvalumab administered is about 60 mg to 900 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of Durvalumab administered is about 5 mg/kg to 15 mg/kg every 2 weeks or every 3 weeks. In some embodiments, the effective amount of Durvalumab administered is about 10 mg/kg every 2 weeks.
在一些实施方案中，每次施用的Durvalumab为约1mg/kg至10mg/kg或含此剂量Durvalumab的制剂。在一些实施方案中，每次施用的Durvalumab为约1mg/kg、约1.2mg/kg、约2mg/kg、约2.4mg/kg、约3mg/kg、约3.6mg/kg、约4mg/kg、约4.8mg/kg、约5mg/kg、约5.5mg/kg、约6mg/kg、约9mg/kg、约10mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量Durvalumab的制剂。In some embodiments, the amount of Durvalumab per administration is from about 1 mg/kg to 10 mg/kg or formulations containing such doses of Durvalumab. In some embodiments, each administration of Durvalumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, About 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 6 mg/kg, about 9 mg/kg, about 10 mg/kg, or a range between any two of these values (inclusive) or where Any value, or formulations containing this dose of Durvalumab.
在一些实施方案中，采用治疗有效量的抗PD-L1抗体和抗OX40抗体分别或者同时施加在患者上。抗PD-L1抗体和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of the anti-PD-L1 antibody and the anti-OX40 antibody are administered to the patient separately or simultaneously. The administration cycles of the anti-PD-L1 antibody and the anti-OX40 antibody can be the same or different.
在一些实施方案中，所述另一个治疗剂为抗TIGIT抗体或抗原结合片段。在一些实施方案中，所述抗TIGIT抗体为Tiragolumab(替瑞利尤单抗)或其生物类似物或ADCC效应增强单抗或去岩藻糖基化单抗，或WO2021043206中描述的抗TIGIT抗体或抗原结合片段。In some embodiments, the other therapeutic agent is an anti-TIGIT antibody or antigen-binding fragment. In some embodiments, the anti-TIGIT antibody is Tiragolumab (tireliumab) or its biosimilar or ADCC effect-enhancing monoclonal antibody or defucosylated monoclonal antibody, or the anti-TIGIT antibody described in WO2021043206 or antigen-binding fragments.
在一些实施方案中，所述抗TIGIT抗体或抗原结合片段包含SEQ ID NO:13所示的HCDR1、SEQ ID NO:14所示的HCDR2、SEQ ID NO:15所示的HCDR3、SEQ ID  NO:16所示的LCDR1、SEQ ID NO:17所示的LCDR2和SEQ ID NO:18所示的LCDR3。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:13, HCDR2 shown in SEQ ID NO:14, HCDR3 shown in SEQ ID NO:15, SEQ ID NO: LCDR1 shown in 16, LCDR2 shown in SEQ ID NO:17, and LCDR3 shown in SEQ ID NO:18.
在一些实施方案中，所述抗TIGIT抗体或抗原结合片段的重链可变区包含SEQ ID NO:19所示的序列，与SEQ ID NO:19所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:19所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或所述抗TIGTI抗体或抗原结合片段的轻链可变区包含SEQ ID NO:20所示的序列，与SEQ ID NO:20所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:20所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。In some embodiments, the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO: 19, which is at least 80% identical to the sequence set forth in SEQ ID NO: 19 sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 19; and/or the light chain variable region of the anti-TIGTI antibody or antigen-binding fragment comprises SEQ ID NO: 20 The sequence shown is a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO:20, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:20.
在一些实施方案中，所述抗TIGIT抗体为抗体h10D8OF或h10D8OFKF，抗体h10D8OF和h10D8OFKF的重链包含如SEQ ID NO:21所示序列，抗体h10D8OF和h10D8OFKF的轻链包含如SEQ ID NO:22所示序列；抗体h10D8OF和h10D8OFKF分别含有两条序列相同的重链和两条序列相同的轻链。In some embodiments, the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF, the heavy chain of antibody h10D8OF and h10D8OFKF comprises the sequence shown in SEQ ID NO: 21, and the light chain of antibody h10D8OF and h10D8OFKF comprises the sequence shown in SEQ ID NO: 22 The sequences are shown; antibodies h10D8OF and h10D8OFKF contain two identical heavy chains and two identical light chains, respectively.
在一些实施方案中，所述抗TIGIT抗体(例如抗体h10D8OFKF)或抗原结合片段的岩藻糖基化水平为0-10％。在一些实施方案中，所述抗TIGIT抗体(例如抗体h10D8OFKF)或抗原结合片段的岩藻糖基化水平为0-5％。在一些实施方案中，所述抗TIGIT抗体(例如抗体h10D8OFKF)或抗原结合片段的岩藻糖基化水平为约0、约0.1％、约0.5％、约0.8％、约1％、约1.3％、约1.6％、约2.1％、2.9％、约3％、约3.3％、3.8％、约4％、约4.2％、约4.3％、约4.6％、约5％，或这些数值中任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，所述抗TIGIT抗体(例如抗体h10D8OFKF)或抗原结合片段没有结合岩藻糖。在一些实施方案中，所述抗TIGIT抗体(例如抗体h10D8OFKF)或抗原结合片段具有增强的ADCC效应(antibody-dependent cell-mediated cytotoxicity)。In some embodiments, the level of fucosylation of the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment is 0-10%. In some embodiments, the level of fucosylation of the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment is 0-5%. In some embodiments, the fucosylation level of the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment is about 0, about 0.1%, about 0.5%, about 0.8%, about 1%, about 1.3% , about 1.6%, about 2.1%, 2.9%, about 3%, about 3.3%, 3.8%, about 4%, about 4.2%, about 4.3%, about 4.6%, about 5%, or any two of these values A range between values (inclusive) or any value in it. In some embodiments, the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment does not bind fucose. In some embodiments, the anti-TIGIT antibody (such as antibody h10D8OFKF) or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
在一些实施方案中，抗TIGIT抗体或抗原结合片段可以通过基因工程在CHO细胞或293细胞中表达，并通过纯化获得；纯化可以采用常规方法进行，例如先离心细胞悬液并收集上清液，再次离心进一步去除杂质。ProteinA亲和柱和离子交换柱等方法可以用于纯化抗体蛋白。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment can be expressed in CHO cells or 293 cells by genetic engineering, and obtained by purification; purification can be performed by conventional methods, such as centrifuging the cell suspension first and collecting the supernatant, Centrifuge again to further remove impurities. Methods such as ProteinA affinity column and ion exchange column can be used to purify antibody protein.
在一些实施方案中，低岩藻糖基化或无岩藻糖基化的抗TIGIT抗体或抗原结合片段由α-(1,6)-岩藻糖基转移酶基因敲除的细胞系表达。在一些实施方案中，抗体h10D8OFKF由α-(1,6)-岩藻糖基转移酶基因敲除的CHO细胞表达。In some embodiments, the hypofucosylated or afucosylated anti-TIGIT antibody or antigen-binding fragment is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line. In some embodiments, antibody h10D8OFKF is expressed by α-(1,6)-fucosyltransferase knockout CHO cells.
在一些实施方案中，抗TIGIT抗体施用的有效量为约9mg至1200mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，抗TIGIT抗体施用的有效量为约0.01mg/kg至20mg/kg每2周或每3 周1次。在一些实施方案中，抗TIGIT抗体施用的有效量为约0.01mg/kg、约0.03mg/kg、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、约10mg/kg、约11mg/kg、约12mg/kg、约13mg/kg、约14mg/kg、约15mg/kg、约16mg/kg、约17mg/kg、约18mg/kg、约19mg/kg、约20mg/kg(或这些数值中的任何两个值之间的范围(包括端点)或其中任何值)每2周或每3周1次。在一些实施方案中，抗TIGIT抗体施用的有效量为约10mg至约900mg每2、3周或4周一次。在一些实施方案中，抗TIGIT抗体施用的有效量为约10mg、约30mg、约100mg、约160mg、约200mg、约242mg、约300mg、约346mg、约400mg、约500mg、约600mg(或这些数值中的任何两个值之间的范围(包括端点)或其中任何值)每2周、3周或4周一次。In some embodiments, the effective amount of anti-TIGIT antibody administered is about 9 mg to 1200 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of anti-TIGIT antibody administered is about 0.01 mg/kg to 20 mg/kg once every 2 weeks or every 3 weeks. In some embodiments, the effective amount of anti-TIGIT antibody administered is about 0.01 mg/kg, about 0.03 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, about 5 mg/kg , about 6mg/kg, about 7mg/kg, about 8mg/kg, about 9mg/kg, about 10mg/kg, about 11mg/kg, about 12mg/kg, about 13mg/kg, about 14mg/kg, about 15mg/kg , about 16 mg/kg, about 17 mg/kg, about 18 mg/kg, about 19 mg/kg, about 20 mg/kg (or the range (including endpoints) between any two of these values or any value therein) per 2 Weekly or every 3 weeks. In some embodiments, an effective amount of an anti-TIGIT antibody administered is about 10 mg to about 900 mg once every 2, 3, or 4 weeks. In some embodiments, an effective amount of an anti-TIGIT antibody administered is about 10 mg, about 30 mg, about 100 mg, about 160 mg, about 200 mg, about 242 mg, about 300 mg, about 346 mg, about 400 mg, about 500 mg, about 600 mg (or these values The range between any two values in (inclusive) or any value therein) every 2, 3, or 4 weeks.
在在一些实施方案中，采用治疗有效量的抗TIGIT抗体和抗OX40抗体分别或者同时施加在患者上。抗TIGIT抗体和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of anti-TIGIT antibody and anti-OX40 antibody are administered to the patient separately or simultaneously. The administration periods of the anti-TIGIT antibody and the anti-OX40 antibody may be the same or different.
在一些实施方案中，所述另一个治疗剂为抗GITR抗体或抗原结合片段。在一些实施方案中，所述抗GITR抗体或抗原结合片段，如CN111918878A所述抗体或抗原结合片段。在一些实施方案中，抗GITR抗体施用的有效量为约0.01mg/kg至20mg/kg每2周、每3周或每4周1次。In some embodiments, the other therapeutic agent is an anti-GITR antibody or antigen-binding fragment. In some embodiments, the anti-GITR antibody or antigen-binding fragment, such as the antibody or antigen-binding fragment described in CN111918878A. In some embodiments, the effective amount of anti-GITR antibody administered is about 0.01 mg/kg to 20 mg/kg every 2 weeks, every 3 weeks, or every 4 weeks.
在一些实施方案中，采用治疗有效量的抗GITR抗体和抗OX40抗体分别或者同时施加在患者上。抗GITR抗体和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of anti-GITR antibody and anti-OX40 antibody are administered to the patient separately or simultaneously. The administration cycles of the anti-GITR antibody and the anti-OX40 antibody may be the same or different.
在一些实施方案中，所述另一个治疗剂为与表皮生长因子受体2(HER2)的细胞外二聚化结构域(亚结构域II)发生特异性结合的单克隆抗体(抗HER2抗体)或抗原结合片段，如帕妥珠单抗。帕妥珠单抗可以通过基因工程在细胞(如CHO)中表达，并通过纯化获得。In some embodiments, the other therapeutic agent is a monoclonal antibody that specifically binds to the extracellular dimerization domain (subdomain II) of epidermal growth factor receptor 2 (HER2) (anti-HER2 antibody) Or antigen-binding fragments such as Pertuzumab. Pertuzumab can be expressed in cells (such as CHO) by genetic engineering and obtained by purification.
在一些实施方案中，所述另一个治疗剂为帕妥珠单抗，帕妥珠单抗包括Perjeta TM或其生物类似物或ADCC效应增强单抗或去岩藻糖基化单抗。在一些实施方案中，帕妥珠单抗施用的有效量为约40mg至900mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，帕妥珠单抗施用的有效量为约初始840mg，之后420mg每3周1次。 In some embodiments, the other therapeutic agent is Pertuzumab, including Perjeta ™ or a biosimilar thereof or an ADCC effect enhancing mAb or an afucosylated mAb. In some embodiments, the effective amount of pertuzumab administered is about 40 mg to 900 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of Pertuzumab administered is about an initial 840 mg followed by 420 mg every 3 weeks thereafter.
在一些实施方案中，每次施用的帕妥珠单抗为约1mg/kg至12mg/kg或含此剂量帕妥珠单抗的制剂。在一些实施方案中，每次施用的帕妥珠单抗为约1mg/kg，约1.2mg/kg，约2mg/kg，约2.4mg/kg，约3mg/kg，约3.6mg/kg，约4mg/kg，约4.8mg/kg，约5mg/kg，约5.5mg/kg，约6mg/kg，约6.9mg/kg，约7mg/kg，约8.4mg/kg，约9 mg/kg，约11mg/kg，约12mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量帕妥珠单抗的制剂。In some embodiments, the amount of Pertuzumab per administration is about 1 mg/kg to 12 mg/kg or a formulation containing such doses of Pertuzumab. In some embodiments, pertuzumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, about 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 6 mg/kg, about 6.9 mg/kg, about 7 mg/kg, about 8.4 mg/kg, about 9 mg/kg, about 11 mg/kg, about 12 mg/kg, or a range between any two of these values (inclusive), or any value therein, or a formulation containing such a dose of Pertuzumab.
在一些实施方案中，采用治疗有效量的帕妥珠单抗和抗OX40抗体分别或者同时施加在患者上。帕妥珠单抗和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of pertuzumab and anti-OX40 antibody are administered to the patient separately or simultaneously. The administration periods of Pertuzumab and anti-OX40 antibody can be the same or different.
在一些实施方案中，所述另一个治疗剂是一种重组人源化免疫球蛋白G1(IgG1)单克隆抗体，可以结合VEGF-A，抑制其与VEGF受体-2(VEGFR2)结合(抗VEGF抗体)，如贝伐珠单抗。贝伐珠单抗可以通过基因工程在细胞(如CHO)中表达，并通过纯化获得。In some embodiments, the other therapeutic agent is a recombinant humanized immunoglobulin G1 (IgG1) monoclonal antibody that binds VEGF-A and inhibits its binding to VEGF receptor-2 (VEGFR2) (anti- VEGF antibody), such as bevacizumab. Bevacizumab can be expressed in cells (such as CHO) by genetic engineering and obtained by purification.
在一些实施方案中，所述另一个治疗剂为贝伐珠单抗，贝伐珠单抗包括
Figure PCTCN2022106326-appb-000001
或其生物类似物，如
Figure PCTCN2022106326-appb-000002
BAT1706，或ADCC效应增强单抗或去岩藻糖基化单抗。 In some embodiments, the other therapeutic agent is bevacizumab, bevacizumab comprising
Figure PCTCN2022106326-appb-000001
or its biosimilars, such as
Figure PCTCN2022106326-appb-000002
BAT1706, or ADCC effect enhancing mAb or defucosylated mAb.
在一些实施方案中，贝伐珠单抗施用的有效量为约50mg至400mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，贝伐珠单抗施用的有效量为约5mg/kg到15mg/kg每2周或每3周1次。在一些实施方案中，贝伐珠单抗施用的有效量为约5mg/kg，约7.5mg/kg，约10mg/kg，或约15mg/kg每2周或每3周1次。在一些实施方案中，贝伐珠单抗施用的有效量为约5mg/kg每2周1次，10mg/kg每2周1次，7.5mg/kg每3周1次，15mg/kg每3周1次。In some embodiments, the effective amount of bevacizumab administered is about 50 mg to 400 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of bevacizumab administered is about 5 mg/kg to 15 mg/kg every 2 weeks or every 3 weeks. In some embodiments, the effective amount of bevacizumab administered is about 5 mg/kg, about 7.5 mg/kg, about 10 mg/kg, or about 15 mg/kg every 2 weeks or every 3 weeks. In some embodiments, the effective amount of bevacizumab administered is about 5 mg/kg every 2 weeks, 10 mg/kg every 2 weeks, 7.5 mg/kg every 3 weeks, 15 mg/kg every 3 weeks 1 time a week.
在一些实施方案中，每次施用的贝伐珠单抗为约1mg/kg至9mg/kg或含此剂量贝伐珠单抗的制剂。在一些实施方案中，每次施用的贝伐珠单抗为约1mg/kg，约1.2mg/kg，约2mg/kg，约2.4mg/kg，约3mg/kg，约3.6mg/kg，约4mg/kg，约4.8mg/kg，约5mg/kg，约5.5mg/kg，约6mg/kg，约6.9mg/kg，约7mg/kg，约8.4mg/kg，约9mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量贝伐珠单抗的制剂。In some embodiments, the amount of bevacizumab per administration is about 1 mg/kg to 9 mg/kg or a formulation containing such doses of bevacizumab. In some embodiments, each administration of bevacizumab is about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6 mg/kg, about 4 mg/kg, about 4.8 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 6 mg/kg, about 6.9 mg/kg, about 7 mg/kg, about 8.4 mg/kg, about 9 mg/kg, or these The range (inclusive) between any two values in a value, or any value therein, or a formulation containing this dose of bevacizumab.
在一些实施方案中，采用治疗有效量的贝伐珠单抗和抗OX40抗体分别或者同时施加在患者上。贝伐珠单抗和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of bevacizumab and anti-OX40 antibody are administered to the patient separately or simultaneously. The administration cycles of bevacizumab and anti-OX40 antibody can be the same or different.
在一些实施方案中，所述另一个治疗剂为是靶向CD20的抗体(抗CD20抗体)，如奥法木单抗、obinutuzumab、CN109096399A所述BAT4306F或CN109096399A所述的BAT4406F。奥法木单抗及obinutuzumab可以通过基因工程在细胞(如CHO)中表达，并通过纯化获得。In some embodiments, the other therapeutic agent is an antibody targeting CD20 (anti-CD20 antibody), such as ofatumumab, obinutuzumab, BAT4306F described in CN109096399A or BAT4406F described in CN109096399A. Ofatumumab and obinutuzumab can be expressed in cells (such as CHO) through genetic engineering and obtained through purification.
在一些实施方案中，所述另一个治疗剂为奥法木单抗，奥法木单抗包括Arzerra TM 或
Figure PCTCN2022106326-appb-000003
其生物类似物或ADCC效应增强单抗或去岩藻糖基化单抗。在一些实施方案中，奥法木单抗施用的有效量为约10mg至2000mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，奥法木单抗施用的有效量为约20mg每周或每月1次。在一些实施方案中，奥法木单抗施用的有效量为约初始300mg，1周后1000mg，之后1000mg每4周或每8周1次。在一些实施方案中，奥法木单抗施用的有效量为约初始300mg，1周后2000mg，之后2000mg每1周或每4周1次。 In some embodiments, the other therapeutic agent is ofatumumab, ofatumumab including Arzerra ™ or
Figure PCTCN2022106326-appb-000003
Its biosimilar or ADCC effect enhancing mAb or afucosylated mAb. In some embodiments, ofatumumab is administered in an effective amount of about 10 mg to 2000 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount ofatumumab administered is about 20 mg weekly or monthly. In some embodiments, the effective amount ofatumumab administered is about 300 mg initially, 1000 mg after 1 week, and 1000 mg every 4 weeks or every 8 weeks thereafter. In some embodiments, the effective amount ofatumumab administered is about 300 mg initially, 2000 mg after 1 week, and 2000 mg every 1 week or every 4 weeks thereafter.
在一些实施方案中，每次施用的奥法木单抗为约0.5mg/kg至18mg/kg或含此剂量奥法木单抗的制剂。在一些实施方案中，每次施用的奥法木单抗为约0.5mg/kg，约1mg/kg，约1.2mg/kg，约2mg/kg，约2.4mg/kg，约3mg/kg，约3.6mg/kg，约4mg/kg，约4.8mg/kg，约5mg/kg，约5.5mg/kg，约6mg/kg，约6.9mg/kg，约7mg/kg，约8.4mg/kg，约9mg/kg，约11mg/kg，约13mg/kg，约14mg/kg，约15mg/kg，约17mg/kg，约18mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量奥法木单抗的制剂。In some embodiments, each administration of ofatumumab is about 0.5 mg/kg to 18 mg/kg or a formulation containing such a dose of ofatumumab. In some embodiments, ofatumumab per administration is about 0.5 mg/kg, about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 6.9mg/kg, about 7mg/kg, about 8.4mg/kg, about 9 mg/kg, about 11 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15 mg/kg, about 17 mg/kg, about 18 mg/kg, or the range between any two of these values (inclusive) or any value therein, or a formulation containing this dose of ofatumumab.
在一些实施方案中，采用治疗有效量的奥法木单抗和抗OX40抗体分别或者同时施加在患者上。奥法木单抗和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of ofatumumab and anti-OX40 antibody are administered to the patient separately or simultaneously. The administration periods of ofatumumab and anti-OX40 antibody can be the same or different.
在一些实施方案中，所述另一个治疗剂为obinutuzumab，包括
Figure PCTCN2022106326-appb-000004
其生物类似物,或ADCC效应增强单抗或去岩藻糖基化单抗。在一些实施方案中，obinutuzumab施用的有效量为约10mg至2000mg每个治疗周期。在一些实施方案中，一个治疗周期为1周、2周、3周、4周、1个月、5周、6周、7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，obinutuzumab施用的有效量为第1天100mg，第2天900mg，第8天、第15天1000mg，之后1000mg每疗程。在一些实施方案中，obinutuzumab施用的有效量为第1天、第8天、第15天各1000mg，之后1000mg每疗程。每疗程可以是1个月或2个月。 In some embodiments, the other therapeutic agent is obinutuzumab, comprising
Figure PCTCN2022106326-appb-000004
Its biosimilar, or ADCC effect enhancing monoclonal antibody or defucosylated monoclonal antibody. In some embodiments, obinutuzumab is administered in an effective amount of about 10 mg to 2000 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including endpoint) or any value in it. In some embodiments, the effective amount of obinutuzumab administered is 100 mg on day 1, 900 mg on day 2, 1000 mg on day 8, day 15, and 1000 mg per cycle thereafter. In some embodiments, the effective amount of obinutuzumab administered is 1000 mg each on Day 1, Day 8, and Day 15, and 1000 mg per course thereafter. Each course of treatment can be 1 month or 2 months.
在一些实施方案中，每次施用的obinutuzumab为约0.5mg/kg至15mg/kg或含此剂量奥法木单抗的制剂。在一些实施方案中，每次施用的奥法木单抗为约0.5mg/kg，约1mg/kg，约1.2mg/kg，约2mg/kg，约2.4mg/kg，约3mg/kg，约3.6mg/kg，约4mg/kg，约4.8mg/kg，约5mg/kg，约5.5mg/kg，约6mg/kg，约6.9mg/kg，约7mg/kg，约8.4mg/kg，约9mg/kg，约11mg/kg，约13mg/kg，约14mg/kg，约15mg/kg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量奥法木单抗的制剂。In some embodiments, the amount of obinutuzumab per administration is from about 0.5 mg/kg to 15 mg/kg or a formulation containing such doses of ofatumumab. In some embodiments, ofatumumab per administration is about 0.5 mg/kg, about 1 mg/kg, about 1.2 mg/kg, about 2 mg/kg, about 2.4 mg/kg, about 3 mg/kg, about 3.6mg/kg, about 4mg/kg, about 4.8mg/kg, about 5mg/kg, about 5.5mg/kg, about 6mg/kg, about 6.9mg/kg, about 7mg/kg, about 8.4mg/kg, about 9 mg/kg, about 11 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15 mg/kg, or the range between any two of these values (including the endpoints) or any value therein, or a dose containing Formulations of falimumab.
在一些实施方案中，采用治疗有效量的obinutuzumab和抗OX40抗体分别或者同 时施加在患者上。obinutuzumab和抗OX40抗体的给药周期可以相同或者不同。In some embodiments, therapeutically effective amounts of obinutuzumab and anti-OX40 antibody are administered to the patient separately or simultaneously. The dosing cycles of obinutuzumab and anti-OX40 antibody can be the same or different.
在一些实施方案中，所述另一个治疗剂为抗HER2抗体药物偶联物(HER2-ADC)或抗Trop2抗体药物偶联物(Trop2-ADC)，例如ado-trastuzumab emtansine(T-DM1)，trastuzumab deruxtecan(DS-8201)，CN103333246B及CN109078181A所述抗体药物偶联物。In some embodiments, the other therapeutic agent is an anti-HER2 antibody drug conjugate (HER2-ADC) or an anti-Trop2 antibody drug conjugate (Trop2-ADC), such as ado-trastuzumab emtansine (T-DM1), Trastuzumab deruxtecan (DS-8201), the antibody drug conjugate described in CN103333246B and CN109078181A.
在一些实施方案中，本发明公开了一种治疗肿瘤或癌症的方法，其包括向有需要的患者施用有效量的抗OX40抗体或抗原结合片段(或制剂)和另一个治疗剂(或制剂)；其中，抗OX40抗体或抗原结合片段的有效量为单次给药约0.6mg至900mg(或含此剂量抗OX40抗体的制剂)。在一些实施方案中，另一个治疗剂为抗PD-1抗体或抗原结合片段。In some embodiments, the present invention discloses a method of treating a tumor or cancer comprising administering to a patient in need thereof an effective amount of an anti-OX40 antibody or antigen-binding fragment (or formulation) and another therapeutic agent (or formulation) ; Wherein, the effective dose of anti-OX40 antibody or antigen-binding fragment is about 0.6 mg to 900 mg in a single administration (or a preparation containing this dose of anti-OX40 antibody). In some embodiments, the other therapeutic agent is an anti-PD-1 antibody or antigen-binding fragment.
在一些实施方案中，所述抗PD-1抗体或抗原结合片段包含SEQ ID NO:25所示的HCDR1、SEQ ID NO:26所示的HCDR2、SEQ ID NO:27所示的HCDR3、SEQ ID NO:28所示的LCDR1、SEQ ID NO:29所示的LCDR2和SEQ ID NO:30所示的LCDR3。In some embodiments, the anti-PD-1 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:25, HCDR2 shown in SEQ ID NO:26, HCDR3 shown in SEQ ID NO:27, SEQ ID LCDR1 shown in NO:28, LCDR2 shown in SEQ ID NO:29 and LCDR3 shown in SEQ ID NO:30.
在一些实施方案中，所述抗PD-1抗体或抗原结合片段的重链可变区包含SEQ ID NO:31所示的序列，与SEQ ID NO:31所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:31所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或所述抗PD-1抗体或抗原结合片段的轻链可变区包含SEQ ID NO:32所示的序列，与SEQ ID NO:32所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:32所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。In some embodiments, the heavy chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence set forth in SEQ ID NO:31, which is at least 80% identical to the sequence set forth in SEQ ID NO:31 or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:31; and/or the light chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises SEQ ID NO:31 The sequence shown in ID NO:32, a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO:32, or having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:32 amino acid sequence.
在一些实施方案中，所述抗PD-1抗体的重链包含如SEQ ID NO:33所示的氨基酸序列，抗PD-1抗体的轻链包含如SEQ ID NO:34所示的氨基酸序列。In some embodiments, the heavy chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:33, and the light chain of the anti-PD-1 antibody comprises the amino acid sequence shown in SEQ ID NO:34.
在一些实施方案中，抗PD-1抗体的有效量为单次给药约50mg至600mg(或含此剂量抗PD-1抗体的制剂)。剂量时间表和给药方式取决于某些患者群中的抗PD-1抗体(或制剂)、抗OX40(或制剂)的获益风险评估和一般临床实践指南。In some embodiments, the effective amount of anti-PD-1 antibody is about 50 mg to 600 mg in a single administration (or a preparation containing such dose of anti-PD-1 antibody). Dosage schedule and mode of administration depend on the benefit-risk assessment of anti-PD-1 antibody (or formulation), anti-OX40 (or formulation) in certain patient populations and general clinical practice guidelines.
在一些实施方案中，患者每个治疗周期内抗OX40抗体(例如抗体M或M-KF)施用的有效量为约0.6mg至900mg的抗OX40抗体(或含此剂量抗OX40抗体的制剂)，患者每个治疗周期内抗PD-1抗体施用的有效量为约50mg至600mg(或含此剂量抗PD-1抗体的制剂)。In some embodiments, the anti-OX40 antibody (such as antibody M or M-KF) is administered to the patient in an effective amount of about 0.6 mg to 900 mg of anti-OX40 antibody (or a preparation containing such a dose of anti-OX40 antibody) per treatment cycle, The effective amount of anti-PD-1 antibody administered to the patient in each treatment cycle is about 50 mg to 600 mg (or a preparation containing this dose of anti-PD-1 antibody).
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量是约0.6mg、约1.8mg、约6mg、约9mg、约10mg、约12mg、约18mg、约20mg、约30mg、约50mg、约60mg、约80mg、约120mg、约180mg、约200mg、约250mg、约290mg、约300mg、约330mg、约360mg、约380mg、约400mg、 约434mg、约480mg、约500mg、约567mg、约580mg、约600mg、约700mg、约800mg、约900mg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量抗OX40抗体的制剂。在一些实施方案中，一个治疗周期为1周至7周给药1次。在一些实施方案中，每个治疗周期内施用抗OX40抗体的有效量是0.6mg至100mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周、约4周、约5周、约6周、约7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，一个治疗周期为约1周、约2周、约3周或约4周。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约100mg至约300mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约300mg至约600mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周、或约4周。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约600mg至约900mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约6mg、约18mg、约50mg、约60mg、约100mg、约100mg、约110mg、约120mg、约130mg、约140mg、约150mg、约160mg、约170mg、约180mg、约190mg、约200mg、约260mg、约300mg、约360mg、约400mg、约430mg、约460mg、约520mg、约600mg、约900mg，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) administered to a patient per treatment cycle is about 0.6 mg, about 1.8 mg, about 6 mg, about 9 mg, about 10 mg, about 12 mg, about 18mg, about 20mg, about 30mg, about 50mg, about 60mg, about 80mg, about 120mg, about 180mg, about 200mg, about 250mg, about 290mg, about 300mg, about 330mg, about 360mg, about 380mg, about 400mg, about 434mg, About 480 mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or a range (including endpoints) between any two of these values or any value therein, or containing this dose of anti Preparations of OX40 Antibodies. In some embodiments, a treatment cycle is administered once from 1 week to 7 weeks. In some embodiments, the effective amount of the anti-OX40 antibody administered in each treatment cycle is 0.6 mg to 100 mg, or a preparation containing this dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or a range between any two of these values (inclusive) or any value therein. In some embodiments, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 100 mg to about 300 mg in each treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, About 3 weeks or about 4 weeks. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 300 mg to about 600 mg in each treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, About 3 weeks, or about 4 weeks. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 600 mg to about 900 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, About 3 weeks or about 4 weeks. In some embodiments, the effective amount of the anti-OX40 antibody administered to the patient per treatment cycle is about 6 mg, about 18 mg, about 50 mg, about 60 mg, about 100 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 260 mg, about 300 mg, about 360 mg, about 400 mg, about 430 mg, about 460 mg, about 520 mg, about 600 mg, about 900 mg, or any of these values A range (inclusive) between any two values, or any value therein, or a formulation comprising such an anti-OX40 antibody dose; wherein a treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约5mg至16mg，或含此剂量抗OX40抗体的制剂；比如约6mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约6mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 5 mg to 16 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 6 mg is administered 1 Second-rate. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 6 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约15mg至40mg，或含此剂量抗OX40抗体的制剂；比如约18mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约18mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) of about 15 mg to 40 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 18 mg is administered 1 Second-rate. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 18 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约100mg至190mg，或含此剂量抗OX40抗体的制剂；比如约180mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约180 mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 100 mg to 190 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 180 mg is administered 1 Second-rate. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 180 mg per treatment cycle, or a preparation containing this dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks week or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约167mg至250mg，或含此剂量抗OX40抗体的制剂；比如约210mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约210mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 167 mg to 250 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 210 mg is administered 1 Second-rate. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 210 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约267mg至354mg，或含此剂量抗OX40抗体的制剂；比如约312mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约312mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 267 mg to 354 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; Second-rate. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 312 mg, or a preparation containing such a dose of anti-OX40 antibody, in each treatment cycle; wherein, a treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约380mg至450mg，或含此剂量抗OX40抗体的制剂；比如约400mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约400mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 380 mg to 450 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 400 mg is administered 1 Second-rate. In some embodiments, the patient is administered with an effective amount of anti-OX40 antibody of about 400 mg in each treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约542mg至643mg，或含此剂量抗OX40抗体的制剂；比如约620mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约620mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 542 mg to 643 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 620 mg is administered 1 Second-rate. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 620 mg, or a preparation containing such a dose of anti-OX40 antibody, in each treatment cycle; wherein, a treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(例如抗体M或M-KF)的有效量为约827mg至940mg，或含此剂量抗OX40抗体的制剂；比如约900mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗OX40抗体的有效量为约900mg，或含此剂量抗OX40抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of an anti-OX40 antibody (e.g., antibody M or M-KF) of about 827 mg to 940 mg per treatment cycle, or a formulation containing such an anti-OX40 antibody; for example, about 900 mg is administered 1 Second-rate. In some embodiments, the patient is administered an effective amount of anti-OX40 antibody of about 900 mg per treatment cycle, or a preparation containing such a dose of anti-OX40 antibody; wherein, one treatment cycle is about 1 week, about 2 weeks, or about 3 weeks or about 4 weeks.
在一些实施方案中，抗OX40抗体(例如抗体M或M-KF)的有效量为约0.6mg至900mg每3周一次。在一些实施方案中，抗OX40抗体的有效量为约0.6mg、约1.8mg、约10mg、约30mg、约60mg、约100mg、约180mg、约200mg、约300mg、约360mg、约400mg、约500mg、约600mg、约700mg、约800mg或约900mg每3周一次。在一些实施方案中，抗OX40抗体的有效量为约6mg、约18mg、约60mg、 约180mg、约360mg约600mg或约900mg每3周一次。In some embodiments, the effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) is about 0.6 mg to 900 mg once every 3 weeks. In some embodiments, the effective amount of an anti-OX40 antibody is about 0.6 mg, about 1.8 mg, about 10 mg, about 30 mg, about 60 mg, about 100 mg, about 180 mg, about 200 mg, about 300 mg, about 360 mg, about 400 mg, about 500 mg , about 600 mg, about 700 mg, about 800 mg, or about 900 mg every 3 weeks. In some embodiments, the effective amount of an anti-OX40 antibody is about 6 mg, about 18 mg, about 60 mg, about 180 mg, about 360 mg, about 600 mg, or about 900 mg once every 3 weeks.
在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量是约50mg、约60mg、约80mg、约120mg、约200mg、约250mg、约290mg、约300mg、约330mg、约380mg、约400mg、约434mg、约480mg、约500mg、约567mg、约580mg、约600mg，或这些数值中任何两个值之间的范围(包括端点)或其中任何值，或含此剂量抗PD-1抗体的制剂。在一些实施方案中，一个治疗周期为1周至7周给药1次。在一些实施方案中，每个治疗周期内施用抗PD-1抗体的有效量是100mg至200mg，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周、约4周、约5周、约6周、约7周，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。在一些实施方案中，一个治疗周期为约1周、约2周、约3周或约4周。在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约200mg至约300mg，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周、或约4周。在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约100mg、约110mg、约120mg、约130mg、约140mg、约150mg、约160mg、约170mg、约180mg、约190mg、约200mg、约300mg，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 50 mg, about 60 mg, about 80 mg, about 120 mg, about 200 mg, about 250 mg, about 290 mg, about 300 mg, about 330 mg, about 380 mg, about 400 mg, about 434 mg, about 480 mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, or a range between any two of these values (including endpoints) or any value therein, or containing this dose of anti-PD -1 Preparation of antibodies. In some embodiments, a treatment cycle is administered once from 1 week to 7 weeks. In some embodiments, the effective amount of anti-PD-1 antibody administered in each treatment cycle is 100 mg to 200 mg, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or a range (including endpoints) between any two of these values, or any value therein. In some embodiments, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 200 mg to about 300 mg in each treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; wherein, one treatment cycle is about 1 week, About 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 300 mg, or the range between any two of these values (including the endpoints) or any value therein, or a preparation containing this dose of anti-PD-1 antibody; wherein, one treatment cycle is about 1 weeks, about 2 weeks, about 3 weeks, or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约45mg至80mg，或含此剂量抗PD-1抗体的制剂；比如约50mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约50mg，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 45 mg to 80 mg per treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; for example, about 50 mg is administered once. In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 50 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约87mg至130mg，或含此剂量抗PD-1抗体的制剂；比如约100mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约100mg，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 87 mg to 130 mg per treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; for example, about 100 mg is administered once. In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 100 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约180mg至230mg，或含此剂量抗PD-1抗体的制剂；比如约200mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约200mg，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 180 mg to 230 mg per treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; for example, about 200 mg is administered once. In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 200 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约267mg至343mg，或含此剂量抗PD-1抗体的制剂；比如约300mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约300mg，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the effective amount of anti-PD-1 antibody administered to the patient is about 267 mg to 343 mg, or a preparation containing this dose of anti-PD-1 antibody; for example, about 300 mg is administered once in each treatment cycle. In some embodiments, the patient is administered with an effective amount of anti-PD-1 antibody of about 300 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约300mg至700mg，或含此剂量抗PD-1抗体的制剂；比如约600mg给药1次。在一些实施方案中，患者每个治疗周期内施用抗PD-1抗体的有效量为约600mg，或含此剂量抗PD-1抗体的制剂；其中，一个治疗周期为约1周、约2周、约3周或约4周。In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 300 mg to 700 mg per treatment cycle, or a preparation containing such a dose of anti-PD-1 antibody; for example, about 600 mg is administered once. In some embodiments, the patient is administered an effective amount of anti-PD-1 antibody of about 600 mg in each treatment cycle, or a preparation containing this dose of anti-PD-1 antibody; wherein, a treatment cycle is about 1 week, about 2 weeks , about 3 weeks or about 4 weeks.
在一些实施方案中，患者每个治疗周期内分别给药一次抗OX40抗体和另一个治疗剂(或给药一次抗OX40抗体和另一个治疗剂的组合物)。在一些实施方案中，每个治疗周期内多次分别给药抗OX40抗体和另一个治疗剂(或抗OX40抗体和另一个治疗剂的组合物)，例如2次、3次、4次或5次。在一些实施方案中，患者每个治疗周期只能给药1次或4次。In some embodiments, the patient is administered an anti-OX40 antibody and another therapeutic agent (or a combination of an anti-OX40 antibody and another therapeutic agent) once per treatment cycle. In some embodiments, the anti-OX40 antibody and another therapeutic agent (or a combination of an anti-OX40 antibody and another therapeutic agent) are administered multiple times, such as 2, 3, 4, or 5 times, separately, per treatment cycle. Second-rate. In some embodiments, the patient can only be dosed 1 or 4 times per treatment cycle.
在一些实施方案中，患者接受一个治疗周期治疗。在一些实施方案中，患者接受多个(例如2个、3个、3个、4个、5个、6个、7个、8个、9个、10个、11个、12次、13个、14个、15个、16个或17个)治疗周期治疗。在一些实施方案中，患者接受治疗直至病症得到缓解而不再需要治疗。In some embodiments, the patient is treated for one treatment cycle. In some embodiments, the patient receives multiple (e.g., 2, 3, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16 or 17) treatment cycles. In some embodiments, the patient is treated until the condition is in remission and no longer requires treatment.
在一些实施方案中，本发明公开了一种用于治疗肿瘤或癌症的方法，所述方法包括：向有需要的患者给予约0.6mg至100mg、约100mg至300mg、约300mg至600mg或约600mg至900mg，比如约0.6mg、约1.8mg、约6mg、约10mg、约18mg、约20mg、约30mg、约100mg、约120mg、约180mg、约200mg、约300mg、约360mg、约600mg或约900mg的抗OX40抗体(例如抗体M或M-KF)，或含此剂量抗OX40抗体的制剂；还向有需要的患者给予约50mg至100mg、约100mg至200mg、约200mg至400mg或约400mg至600mg，比如约100mg、约120mg、约150mg、约200mg或约300mg的抗PD-1抗体，或含此剂量抗PD-1抗体的制剂。在一些实施方案中，患者接受单剂量抗约200mg抗体的治疗，以及单剂量抗PD-1抗体的治疗。在一些实施方案中，患者接受单剂量抗OX40抗体和抗PD-1抗体组合物的治疗。In some embodiments, the present invention discloses a method for treating tumor or cancer, said method comprising: administering about 0.6 mg to 100 mg, about 100 mg to 300 mg, about 300 mg to 600 mg, or about 600 mg to a patient in need thereof Up to 900 mg, such as about 0.6 mg, about 1.8 mg, about 6 mg, about 10 mg, about 18 mg, about 20 mg, about 30 mg, about 100 mg, about 120 mg, about 180 mg, about 200 mg, about 300 mg, about 360 mg, about 600 mg, or about 900 mg Anti-OX40 antibody (such as antibody M or M-KF), or preparations containing such doses of anti-OX40 antibody; also administer about 50 mg to 100 mg, about 100 mg to 200 mg, about 200 mg to 400 mg, or about 400 mg to 600 mg to patients in need , such as about 100 mg, about 120 mg, about 150 mg, about 200 mg or about 300 mg of anti-PD-1 antibody, or a preparation containing such a dose of anti-PD-1 antibody. In some embodiments, the patient is treated with a single dose of the anti-about 200 mg antibody, and a single dose of the anti-PD-1 antibody. In some embodiments, the patient is treated with a single dose of the anti-OX40 antibody and anti-PD-1 antibody composition.
在一些实施方案中，每3周一次给药抗OX40抗体和抗PD-1抗体。In some embodiments, the anti-OX40 antibody and the anti-PD-1 antibody are administered every 3 weeks.
在一些实施方案中，单剂量给药后，患者的症状得到缓解。在一些实施方案中，单剂量给药后，患者的症状未得到预期缓解，再对患者分别给药约0.6mg至900mg抗OX40抗体和约50mg至600mg抗PD-1抗体。In some embodiments, following a single dose, the patient's symptoms are relieved. In some embodiments, after a single dose of administration, the patient's symptoms are not relieved as expected, and then about 0.6 mg to 900 mg of the anti-OX40 antibody and about 50 mg to 600 mg of the anti-PD-1 antibody are administered to the patient.
在一些实施方案中，抗OX40抗体为抗体M或M-KF；抗PD-1抗体为抗体A。In some embodiments, the anti-OX40 antibody is Antibody M or M-KF; the anti-PD-1 antibody is Antibody A.
在一些实施方案中，本发明公开了一种用于治疗肿瘤或癌症的方法，所述方法包括向有需要的患者每3周一次给予3mg/kg到10mg/kg的抗OX40抗体M-KF和300mg到600mg的抗PD-1抗体A。在一些实施方案中，所述方法包括向有需要的患者每3周一次给予3mg/kg，5mg/kg，6mg/kg，或10mg/kg的抗体M-KF和300mg的抗体 A。在一些实施方案中，所述方法包括向有需要的患者每3周一次给予3mg/kg，5mg/kg，6mg/kg,或10mg/kg的抗OX40抗体M-KF和600mg的抗PD-1抗体A。在一些实施方案中，所述方法包括在向该患者给予抗体M-KF和抗体A的3周之前先向该患者给予一次同等量的抗体M-KF。In some embodiments, the present invention discloses a method for treating a tumor or cancer, the method comprising administering 3 mg/kg to 10 mg/kg of the anti-OX40 antibody M-KF and 300mg to 600mg of anti-PD-1 antibody A. In some embodiments, the method comprises administering 3 mg/kg, 5 mg/kg, 6 mg/kg, or 10 mg/kg of Antibody M-KF and 300 mg of Antibody A to a patient in need thereof once every 3 weeks. In some embodiments, the method comprises administering 3 mg/kg, 5 mg/kg, 6 mg/kg, or 10 mg/kg of anti-OX40 antibody M-KF and 600 mg of anti-PD-1 once every 3 weeks to a patient in need thereof Antibody A. In some embodiments, the method comprises administering to the patient an equivalent amount of Antibody M-KF once 3 weeks prior to administering Antibody M-KF and Antibody A to the patient.
在一些实施方案中，抗OX40抗体(或制剂)、抗PD-1抗体(或制剂)是通过皮下(s.c.)注射、腹膜内(i.p.)注射、肠胃外注射、动脉内注射或静脉内(i.v.)注射等方式进行给药。在一些实施方案中，抗OX40抗体(或制剂)、抗PD-1抗体(或制剂)是输液方式进行给药。在一些实施方案中，抗OX40抗体(或制剂)、抗PD-1抗体(或制剂)是推注方式进行给药。In some embodiments, the anti-OX40 antibody (or formulation), anti-PD-1 antibody (or formulation) is injected subcutaneously (s.c.), intraperitoneally (i.p.), parenterally, intraarterially, or intravenously (i.v. ) administration by way of injection or the like. In some embodiments, the anti-OX40 antibody (or preparation) and anti-PD-1 antibody (or preparation) are administered by infusion. In some embodiments, the anti-OX40 antibody (or preparation) and anti-PD-1 antibody (or preparation) are administered by bolus injection.
在一些实施方案中，抗OX40抗体(或制剂)、抗PD-1抗体(或制剂)是通过静脉内(i.v.)输液方式进行给药。在一些实施方案中，静脉内输液持续时间为约50分钟、约55分钟、约60分钟、约65分钟、约70分钟、约75分钟、约81分钟、约87分钟、约90分钟、约95分钟，或这些数值中任何两个值之间的范围(包括端点)或其中任何值。In some embodiments, the anti-OX40 antibody (or formulation), anti-PD-1 antibody (or formulation) is administered by intravenous (i.v.) infusion. In some embodiments, the duration of the intravenous infusion is about 50 minutes, about 55 minutes, about 60 minutes, about 65 minutes, about 70 minutes, about 75 minutes, about 81 minutes, about 87 minutes, about 90 minutes, about 95 minutes minutes, or the range between any two of these values (inclusive), or any value therein.
附图说明Description of drawings
图1示抗OX40抗体M-KF抑制肿瘤的生长；纵坐标表示肿瘤体积。Figure 1 shows that the anti-OX40 antibody M-KF inhibits the growth of tumor; the ordinate indicates the tumor volume.
术语the term
除非另作说明，否则下列的每一个术语应当具有下文所述的含义。Unless otherwise specified, each of the following terms shall have the meaning set forth below.
定义definition
应当注意的是，术语“一种”实体是指一种或多种该实体，例如“一种抗体”应当被理解为一种或多种抗体，因此，术语“一种”(或“一个”)、“一种或多种”和“至少一种”可以在本文中互换使用。It should be noted that the term "an" entity refers to one or more such entities, for example "an antibody" should be understood as one or more antibodies, therefore, the term "a" (or "an" ), "one or more" and "at least one" may be used interchangeably herein.
本文所用的术语“包含”或“包括”意味着组合物和方法等包括所列举的元素，例如组份或步骤，但不排除其它。“基本上由……组成”意味着组合物和方法排除对组合的特征有根本影响的其它元素，但不排除对组合物或方法无本质上影响的元素。“由……组成”意味着排除未特别列举的元素。As used herein, the term "comprising" or "comprising" means that compositions, methods, etc. include the listed elements, such as components or steps, but not exclude others. "Consisting essentially of" means that the compositions and methods exclude other elements that substantially affect the characteristics of the combination, but do not exclude elements that do not substantially affect the composition or method. "Consisting of" means excluding elements not specifically recited.
术语“多肽”旨在涵盖单数的“多肽”以及复数的“多肽”，并且是指由通过酰胺键(也称为肽键)线性连接的氨基酸单体构成的分子。术语“多肽”是指两个或更多个氨基酸的任何单条链或多条链，并且不涉及产物的特定长度。因此，“多肽”的定义中包括肽、二肽、三肽、寡肽、“蛋白质”、“氨基酸链”或用于指两个或多个氨基酸链的任何其他术语，并且术语“多肽”可以用来代替上述任何一个术语，或者与上述任何一个术语交 替使用。术语“多肽”也意在指多肽表达后修饰的产物，包括但不限于糖基化、乙酰化、磷酸化、酰胺化、通过已知的保护/封闭基团衍生化、蛋白水解切割或非天然发生的氨基酸修饰。多肽可以源自天然生物来源或通过重组技术产生，但其不必从指定的核酸序列翻译所得，它可能以包括化学合成的任何方式产生。The term "polypeptide" is intended to encompass the singular as well as the plural "polypeptides" and refers to a molecule composed of amino acid monomers linked linearly by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any chain or chains of two or more amino acids, and does not refer to a specific length of the product. Thus, the definition of "polypeptide" includes peptide, dipeptide, tripeptide, oligopeptide, "protein", "amino acid chain" or any other term used to refer to a chain of two or more amino acids, and the term "polypeptide" may Used in place of, or interchangeably with, any of the above terms. The term "polypeptide" is also intended to refer to the products of post-expression modifications of the polypeptide, including but not limited to glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, or non-natural Amino acid modifications that occur. A polypeptide may be derived from natural biological sources or produced by recombinant techniques, but it need not be translated from a specified nucleic acid sequence, it may be produced by any means including chemical synthesis.
“氨基酸”是指既含氨基又含羧基的有机化合物，比如α-氨基酸，其可直接或以前体的形式由核酸编码。单个氨基酸由三个核苷酸(所谓的密码子或碱基三联体)组成的核酸编码。每一个氨基酸由至少一个密码子编码。相同氨基酸由不同密码子编码称为“遗传密码的简并性”。氨基酸包括天然氨基酸和非天然氨基酸。天然氨基酸包括丙氨酸(三字母代码：ala，一字母代码：A)、精氨酸(arg，R)、天冬酰胺(asn，N)、天冬氨酸(asp，D)、半胱氨酸(cys，C)、谷氨酰胺(gln，Q)、谷氨酸(glu，E)、甘氨酸(gly，G)、组氨酸(his，H)、异亮氨酸(ile，I)、亮氨酸(leu，L)、赖氨酸(lys，K)、甲硫氨酸(met，M)、苯丙氨酸(phe，F)、脯氨酸(pro，P)、丝氨酸(ser，S)、苏氨酸(thr，T)、色氨酸(trp，W)、酪氨酸(tyr，Y)和缬氨酸(val，V)。"Amino acid" refers to an organic compound containing both amino and carboxyl groups, such as an α-amino acid, which can be encoded by a nucleic acid directly or in the form of a precursor. A single amino acid is encoded by a nucleic acid consisting of three nucleotides (so-called codons or base triplets). Each amino acid is encoded by at least one codon. The fact that the same amino acid is encoded by different codons is called "degeneracy of the genetic code". Amino acids include natural amino acids and unnatural amino acids. Natural amino acids include alanine (three-letter code: ala, one-letter code: A), arginine (arg, R), asparagine (asn, N), aspartic acid (asp, D), cysteine amino acid (cys, C), glutamine (gln, Q), glutamic acid (glu, E), glycine (gly, G), histidine (his, H), isoleucine (ile, I ), leucine (leu, L), lysine (lys, K), methionine (met, M), phenylalanine (phe, F), proline (pro, P), serine (ser, S), threonine (thr, T), tryptophan (trp, W), tyrosine (tyr, Y) and valine (val, V).
“保守氨基酸取代”是指一个氨基酸残基被另一个含有化学性质(例如电荷或疏水性)相似的侧链(R基团)的氨基酸残基所取代。一般而言，保守氨基酸取代不大会在实质上改变蛋白质的功能性质。含有化学性质相似侧链的氨基酸类别的实例包括：1)脂族侧链：甘氨酸、丙氨酸、缬氨酸、亮氨酸和异亮氨酸；2)脂族羟基侧链：丝氨酸和苏氨酸；3)含酰胺的侧链：天冬酰胺和谷氨酰胺；4)芳族侧链：苯丙氨酸、酪氨酸和色氨酸；5)碱性侧链：赖氨酸、精氨酸和组氨酸；6)酸性侧链：天冬氨酸和谷氨酸。A "conservative amino acid substitution" refers to the replacement of one amino acid residue with another amino acid residue containing a side chain (R group) of similar chemical properties (eg, charge or hydrophobicity). In general, conservative amino acid substitutions are unlikely to substantially alter the functional properties of a protein. Examples of classes of amino acids that contain chemically similar side chains include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, Arginine and histidine; 6) acidic side chains: aspartic acid and glutamic acid.
“VL、VH的保守氨基酸取代”的氨基酸数目可为约1个、约2个、约3个、约4个、约5个、约6个、约8个、约9个、约10个、约11个、约13个、约14个、约15个保守氨基酸取代，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。“重链或轻链的保守氨基酸取代”的氨基酸数目可为约1个、约2个、约3个、约4个、约5个、约6个、约8个、约9个、约10个、约11个、约13个、约14个、约15个、约18个、约19个、约22个、约24个、约25个、约29个、约31个、约35个、约38个、约41个、约45个保守氨基酸取代，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。The number of amino acids in the "conservative amino acid substitution of VL, VH" can be about 1, about 2, about 3, about 4, about 5, about 6, about 8, about 9, about 10, About 11, about 13, about 14, about 15 conservative amino acid substitutions, or a range between any two of these values (inclusive), or any value therein. The number of amino acids for "conservative amino acid substitutions in the heavy or light chain" can be about 1, about 2, about 3, about 4, about 5, about 6, about 8, about 9, about 10 about 11, about 13, about 14, about 15, about 18, about 19, about 22, about 24, about 25, about 29, about 31, about 35, About 38, about 41, about 45 conservative amino acid substitutions, or a range between any two of these values (inclusive), or any value therein.
术语“编码”应用于多聚核苷酸时，是指被称为“编码”多肽的多聚核苷酸，在其天然状态或当通过本领域技术人员公知的方法操作时，经转录和/或翻译可以产生该多肽和/或其片段。The term "encoding" when applied to a polynucleotide refers to a polynucleotide which is said to "encode" a polypeptide which, in its native state or when manipulated by methods well known to those skilled in the art, is transcribed and/or Or translation may result in the polypeptide and/or fragments thereof.
本发明公开的抗体、抗原结合片段或衍生物包括但不限于多克隆、单克隆、多特异性、全人源、人源化、灵长类化、嵌合抗体、单链抗体、表位结合片段(例如Fab、 Fab'、F(ab') 2和scFv)。 Antibodies, antigen-binding fragments or derivatives disclosed in the present invention include but are not limited to polyclonal, monoclonal, multispecific, fully human, humanized, primatized, chimeric antibodies, single chain antibodies, epitope binding Fragments (eg Fab, Fab', F(ab') 2 and scFv).
术语“重组”涉及多肽或多聚核苷酸，意指天然不存在的多肽或多聚核苷酸的形式，不受限制的实施例可以通过组合产生通常并不存在的多聚核苷酸或多肽。The term "recombinant" refers to polypeptides or polynucleotides, meaning forms of polypeptides or polynucleotides that do not occur in nature, non-limiting examples may be produced by combination of polynucleotides or polynucleotides that do not normally exist or peptide.
“同源性”或“同一性”或“相似性”是指两个肽之间或两个核酸分子之间的序列相似性。可以通过比较每个序列中可以比对的位置来确定同源性。当被比较的序列中的位置被相同的碱基或氨基酸占据时，则分子在该位置是同源的。序列之间的同源程度是由序列共有的匹配或同源位置的数目组成的一个函数。"Homology" or "identity" or "similarity" refers to the sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing the alignable positions in each sequence. When a position in the sequences being compared is occupied by the same base or amino acid, then the molecules are homologous at that position. The degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences.
“至少80％同一性”为约80％同一性、约81％同一性、约82％同一性、约83％同一性、约85％同一性、约86％同一性、约87％同一性、约88％同一性、约90％同一性、约91％同一性、约92％同一性、约94％同一性、约95％同一性、约98％同一性、约99％同一性，或这些数值中的任何两个值之间的范围(包括端点)或其中任何值。"At least 80% identity" is about 80% identity, about 81% identity, about 82% identity, about 83% identity, about 85% identity, about 86% identity, about 87% identity, About 88% identity, about 90% identity, about 91% identity, about 92% identity, about 94% identity, about 95% identity, about 98% identity, about 99% identity, or these A range (inclusive) between any two values in Numeric or any value therein.
多聚核苷酸或多聚核苷酸序列(或多肽或抗体序列)与另一序列有具有一定百分比(例如90％、95％、98％或者99％)的“同一性”或“序列同一性”是指当序列比对时，所比较的两个序列中该百分比的碱基(或氨基酸)相同。可以使用目测或本领域已知的软件程序来确定该比对同一性百分比或序列同一性，比如Ausubel et al.eds.(2007)在Current Protocols in Molecular Biology中所述的软件程序。优选使用默认参数进行比对。其中一种比对程序是使用默认参数的BLAST，例如BLASTN和BLASTP，两者使用下列默认参数：Geneticcode＝standard；filter＝none；strand＝both；cutoff＝60；expect＝10；Matrix＝BLOSUM62；Descriptions＝50sequences；sortby＝HIGHSCORE；Databases＝non-redundant；GenBank+EMBL+DDBJ+PDB+GenBankCDStranslations+SwissProtein+SPupdate+PIR。生物学上等同的多聚核苷酸是具有上述指定百分比的同一性并编码具有相同或相似生物学活性的多肽的多聚核苷酸。A polynucleotide or polynucleotide sequence (or polypeptide or antibody sequence) has a certain percentage (eg, 90%, 95%, 98%, or 99%) of "identity" or "sequence identity" with another sequence. "Sex" means that when the sequences are aligned, the percentage of bases (or amino acids) in the two sequences being compared are identical. This alignment percent identity or sequence identity can be determined using visual inspection or software programs known in the art, such as those described by Ausubel et al.eds. (2007) in Current Protocols in Molecular Biology. It is preferred to use the default parameters for the alignment. One such alignment program is BLAST with default parameters, such as BLASTN and BLASTP, both of which use the following default parameters: Geneticcode=standard; filter=none; strand=both; cutoff=60; expect=10; Matrix=BLOSUM62; Descriptions =50sequences; sortby=HIGHSCORE; Databases=non-redundant; GenBank+EMBL+DDBJ+PDB+GenBankCDStranslations+SwissProtein+SPupdate+PIR. Biologically equivalent polynucleotides are polynucleotides that share the above indicated percentages of identity and encode a polypeptide having the same or similar biological activity.
“抗体”、“抗原结合片段”是指特异性识别和结合抗原的多肽或多肽复合物。抗体可以是完整的抗体及其任何抗原结合片段或其单链。因此术语“抗体”包括分子中含有具有与抗原结合的生物学活性的免疫球蛋白分子的至少一部分的任何蛋白质或肽。抗体和抗原结合片段包括但不局限重链或轻链或其配体结合部分的互补决定区(CDR)、重链可变区(VH)、轻链可变区(VL)、重链恒定区(CH)、轻链恒定区(CL)、框架区(FR)或其任何部分，或结合蛋白的至少一部分。CDR区包括轻链的CDR区(L CDR1-3)和重链的CDR区(HCDR1-3)。"Antibody" and "antigen-binding fragment" refer to a polypeptide or polypeptide complex that specifically recognizes and binds to an antigen. Antibodies can be whole antibodies and any antigen-binding fragments thereof or single chains thereof. The term "antibody" thus includes any protein or peptide whose molecule contains at least a portion of an immunoglobulin molecule that has the biological activity to bind an antigen. Antibodies and antigen-binding fragments include, but are not limited to, complementarity determining regions (CDRs), heavy chain variable regions (VH), light chain variable regions (VL), heavy chain constant regions of heavy or light chains or ligand-binding portions thereof (CH), light chain constant region (CL), framework region (FR) or any portion thereof, or at least a portion of a binding protein. The CDR regions include the CDR regions of the light chain (LCDR1-3) and the CDR regions of the heavy chain (HCDR1-3).
术语“抗体”包括可以在生物化学上区分的各种广泛种类的多肽。本领域技术人员将会理解，重链的类别包括gamma、mu、alpha、delta或epsilon(γ、μ、α、δ、ε)，其中还有一些亚类(例如γ1-γ4)。该链的性质决定了抗体的“种类”分别为IgG、IgM、 IgA、IgG或IgE。免疫球蛋白亚类(同种型)，例如IgG1、IgG2、IgG3、IgG4、IgG5等已被充分表征并且赋予的功能特异性也已知。所有的免疫球蛋白种类都在本发明公开的保护范围内。在一些实施方案中，免疫球蛋白分子为IgG种类。The term "antibody" includes a wide variety of polypeptides that can be distinguished biochemically. Those skilled in the art will understand that the classes of heavy chains include gamma, mu, alpha, delta or epsilon (γ, μ, α, δ, ε), with some subclasses (eg γ1-γ4). The nature of this chain determines the "class" of the antibody as IgG, IgM, IgA, IgG or IgE, respectively. The immunoglobulin subclasses (isotypes), eg, IgGl, IgG2, IgG3, IgG4, IgG5, etc., are well characterized and the functional specificities conferred are also known. All immunoglobulin classes are within the scope of the present disclosure. In some embodiments, the immunoglobulin molecule is of the IgG class.
轻链可以分为kappa(κ)或lambda(λ)。每个重链可以与κ或λ轻链结合。一般来说，当由杂交瘤，B细胞或基因工程宿主细胞生产免疫球蛋白时，其轻链和重链通过共价键结合，两条重链的“尾巴”部分通过共价二硫键或非共价键结合。在重链中，氨基酸序列从Y构型的叉状末端的N末端延伸至每条链底部的C末端。免疫球蛋白κ轻链可变区为Vκ；免疫球蛋白λ轻链可变区为V λ。 Light chains can be classified as kappa (κ) or lambda (λ). Each heavy chain can be associated with a kappa or lambda light chain. Generally, when immunoglobulins are produced by hybridomas, B cells, or genetically engineered host cells, their light and heavy chains are joined by covalent bonds, and the "tail" portions of the two heavy chains are linked by covalent disulfide bonds or non-covalent bonding. In the heavy chains, the amino acid sequence extends from the N-terminus at the forked end of the Y configuration to the C-terminus at the bottom of each chain. The variable region of the immunoglobulin kappa light chain is Vκ; the variable region of the immunoglobulin lambda light chain is Vλ.
轻链和重链都分成结构和功能同源性的区域。术语“恒定的”和“可变的”根据功能被使用。轻链可变区(VL)和重链可变区(VH)链决定了抗原识别和特异性。轻链的恒定区(CL)和重链的恒定区(CH)赋予重要的生物学性质，如分泌、经胎盘移动、Fc受体结合、补体结合等。按照惯例，恒定区的编号随着它们变得更远离抗体的抗原结合位点或氨基末端而增加。N端部分是可变区，C端部分是恒定区；CH3和CL结构域实际上分别包含重链和轻链的羧基端。Both light and heavy chains are divided into regions of structural and functional homology. The terms "constant" and "variable" are used according to function. The variable light (VL) and variable heavy (VH) chains determine antigen recognition and specificity. The constant region (CL) of the light chain and the constant region (CH) of the heavy chain confer important biological properties such as secretion, transplacental movement, Fc receptor binding, complement fixation, etc. By convention, the numbering of constant regions increases as they become farther away from the antigen-binding site or amino terminus of the antibody. The N-terminal portion is the variable region and the C-terminal portion is the constant region; the CH3 and CL domains actually comprise the carboxy-terminal ends of the heavy and light chains, respectively.
在本领域中使用和/或接受的术语有两个或多个定义的情况下，除非明确地对立指出，否则本文使用的术语的定义包括所有这些含义。一个具体的例子是使用“互补决定区”(“CDR”)一词来描述在重链和轻链多肽的可变区内发现的非连续的抗原结合位点。这一特定区域在Kabat et al.,U.S.Dept.of Health and Human Services,Sequences of Proteins of Immunological Interest(1983)和Chothia等在J.Mol.Biol.196:901-917(1987)有相关描述，其通过引用全部并入本文。Where there are two or more definitions for a term used and/or accepted in the art, the definition of the term used herein includes all such meanings unless clearly indicated to the contrary. A specific example is the use of the term "complementarity determining regions" ("CDR") to describe the non-contiguous antigen binding sites found within the variable regions of heavy and light chain polypeptides. This specific region is described in Kabat et al., U.S.Dept.of Health and Human Services, Sequences of Proteins of Immunological Interest (1983) and Chothia et al. in J.Mol.Biol.196:901-917 (1987), It is hereby incorporated by reference in its entirety.
根据Kabat和Chothia定义的CDR包括相互比较时的氨基酸残基的重叠或子集。尽管如此，应用任一定义来指代抗体或其变体的CDR都在本发明范围内。包含特定CDR的确切残基编号将根据CDR的序列和大小而变化。本领域技术人员通常可以根据抗体的可变区氨基酸序列确定出CDR包含哪些特定的残基。CDRs as defined by Kabat and Chothia include overlapping or subsets of amino acid residues when compared to each other. Nevertheless, it is within the scope of the invention to use either definition to refer to the CDRs of an antibody or variant thereof. The exact residue numbers comprising a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in the art can generally determine which specific residues are included in the CDRs based on the amino acid sequence of the variable region of the antibody.
Kabat等人还定义了适用于任何抗体的可变区序列的编号系统。本领域普通技术人员可以不依赖于序列本身以外的其他实验数据将该“Kabat编号”系统应用到任何可变区序列。“Kabat编号”是指由Kabat et al.,U.S.Dept.of Health and Human Services在“Sequence of Proteinsof Immunological Interest”(1983)提出的编号系统。抗体还可以用EU或Chothia编号系统。Kabat et al. also defined a numbering system applicable to the variable region sequences of any antibody. One of ordinary skill in the art can apply this "Kabat numbering" system to any variable region sequence independently of other experimental data other than the sequence itself. "Kabat numbering" refers to the numbering system proposed by Kabat et al., U.S. Dept. of Health and Human Services in "Sequence of Proteins of Immunological Interest" (1983). Antibodies can also use the EU or Chothia numbering system.
本发明中术语“抗体药物偶联物”或“ADC”是指与一个或多个化学药物(其可以任选地是治疗剂或细胞毒性剂)化学连接的抗体或其抗原结合片段。在一些实施方案中，ADC包括抗体、细胞毒性或治疗药物和使得药物能够与抗体连接或偶联的接头。 ADC通常具有与抗体偶联的1、2、3、4、5、6、7、8、9或10个数的药物。可以包括在ADC中的药物有但不限于：有丝分裂抑制剂、抗肿瘤抗生素、免疫调节剂、基因治疗的载体、烷化剂、抗血管生成剂、抗代谢物、含硼试剂、化疗保护剂、激素、抗激素剂、皮质类固醇、光活性治疗剂、寡核苷酸、放射性核素试剂、拓扑异构酶抑制剂、酪氨酸激酶抑制剂和放射致敏剂。在一些实施方案中，包括在ADC中的药物可以是类美登素药物。在一些实施方案中，包括在ADC中的药物可以是如本申请所述的如式Ⅰ所示的化合物或其药学上可接受的盐。在一些实施方案中，在ADC中，抗体通过自身半胱氨酸或疏基化的氨基酸如疏基化赖氨酸，形成二硫键，与药物偶联。The term "antibody drug conjugate" or "ADC" in the present invention refers to an antibody or antigen-binding fragment thereof chemically linked to one or more chemical drugs (which may optionally be therapeutic or cytotoxic agents). In some embodiments, an ADC includes an antibody, a cytotoxic or therapeutic drug, and a linker that enables linking or conjugation of the drug to the antibody. ADCs typically have 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 drug conjugated to the antibody. Drugs that can be included in ADCs are, but not limited to: mitotic inhibitors, antitumor antibiotics, immunomodulators, vectors for gene therapy, alkylating agents, antiangiogenic agents, antimetabolites, boron-containing agents, chemoprotectants, Hormones, antihormones, corticosteroids, photoactive therapeutics, oligonucleotides, radionuclide agents, topoisomerase inhibitors, tyrosine kinase inhibitors, and radiosensitizers. In some embodiments, the drug included in the ADC may be a maytansinoid drug. In some embodiments, the drug included in the ADC may be a compound represented by formula I as described in this application or a pharmaceutically acceptable salt thereof. In some embodiments, in an ADC, the antibody is conjugated to the drug via a self-cysteine or a thiolated amino acid, such as a thiolated lysine, forming a disulfide bond.
“治疗”是指治疗性治疗和预防性或防治性措施，其目的是预防、减缓、改善和停止不良的生理改变或紊乱，例如疾病的进程，包括但不限于以下无论是可检测还是不可检测的结果，症状的缓解、疾病程度的减小、疾病状态的稳定(即不恶化)、疾病进展的延迟或减缓、疾病状态的改善或缓和，减轻或消失(无论是部分还是全部)、延长与不接受治疗时预期的生存期限等。需要治疗的患者包括已经患有病症或紊乱的患者，容易患有病症或紊乱的患者，或者需要预防该病症或紊乱的患者，可以或预期从施用本发明公开的抗体或组合物用于检测、诊断过程和/或治疗中受益的患者。"Treatment" means therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent, slow down, ameliorate and stop undesirable physiological changes or disorders, such as the progression of disease, including but not limited to the following whether detectable or undetectable Relief of symptoms, reduction of disease extent, stabilization of disease state (i.e. not worsening), delay or slowing of disease progression, improvement or palliation of disease state, alleviation or disappearance (whether partial or total), prolongation and Expected survival without treatment, etc. Patients in need of treatment include those who already have a condition or disorder, are susceptible to a condition or disorder, or are in need of prevention of the condition or disorder, and can or are expected to benefit from the administration of an antibody or composition disclosed herein for detection, Patients who benefit from the diagnostic process and/or treatment.
“患者”指需要诊断、预后或治疗的任何哺乳动物，包括人类、狗、猫、豚鼠、兔子、大鼠、小鼠、马、牛等。在一些实施方案中，患者为人。"Patient" refers to any mammal in need of diagnosis, prognosis, or treatment, including humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, and the like. In some embodiments, the patient is a human.
“约”指相关技术领域技术人员容易知道的相应数值的常规误差范围。在一些实施方式中，本文中提到“约”指所描述的数值以及其±10％、±5％或±1％的范围。"About" refers to the usual error range for the corresponding value readily known to those skilled in the relevant art. In some embodiments, reference to "about" herein refers to the described numerical value and the range of ±10%, ±5% or ±1%.
“有效量”是指活性化合物或药剂的量，其能引起组织、系统、动物、个体或人类的生物学或医学反应；有效量由研究人员、兽医、医生或其他临床医生寻求的。"Effective amount" means that amount of an active compound or agent that elicits a biological or medical response in a tissue, system, animal, individual or human; an effective amount is sought by a researcher, veterinarian, physician or other clinician.
如本文所用，短语“有需要”是指已将患者鉴定为需要特定方法或治疗。在一些实施例中，可以通过任何诊断方式进行识别。在本文描述的任何方法和治疗中，患者可能需要。As used herein, the phrase "in need thereof" means that a patient has been identified as being in need of a particular method or treatment. In some embodiments, identification may be by any diagnostic means. A patient may in need thereof during any of the methods and treatments described herein.
可以按常规方法根据本文所述抗体氨基酸序列设计合成编码抗体的DNA，将其置入表达载体中，然后转染宿主细胞，在培养基中培养被转染的宿主细胞产生单克隆抗体。在一些实施方案中，表达抗体载体包括至少一个启动子元件，抗体编码序列，转录终止信号和polyA尾。其他元件包括增强子，Kozak序列及插入序列两侧RNA剪接的供体和受体位点。可以通过SV40的前期和后期启动子，来自逆转录病毒的长末端重复序列如RSV、HTLV1、HIVI及巨细胞病毒的早期启动子来获得高效的转录，也可应用其它一些细胞的启动子如肌动蛋白启动子。合适的表达载体可包括pIRES1neo，pRetro-Off，pRetro-On，PLXSN，或者pLNCX，pcDNA3.1(+/-)，pcDNA/Zeo(+/-)， pcDNA3.1/Hygro(+/-)，PSVL，PMSG，pRSVcat，pSV2dhfr，pBC12MI和pCS2等。常使用的哺乳动物细胞包括293细胞，Cos1细胞，Cos7细胞，CV1细胞，鼠L细胞和CHO细胞等。The DNA encoding the antibody can be designed and synthesized according to the amino acid sequence of the antibody described herein according to conventional methods, placed into an expression vector, and then transfected into a host cell, and the transfected host cell is cultured in a culture medium to produce a monoclonal antibody. In some embodiments, an antibody expression vector includes at least one promoter element, an antibody coding sequence, a transcription termination signal, and a polyA tail. Other elements include enhancers, Kozak sequences, and donor and acceptor sites for RNA splicing flanking the inserted sequence. High-efficiency transcription can be obtained through the early and late promoters of SV40, long terminal repeats from retroviruses such as RSV, HTLV1, HIVI, and early promoters of cytomegalovirus, and other cellular promoters such as muscle Kinetin promoter. Suitable expression vectors may include pIRES1neo, pRetro-Off, pRetro-On, PLXSN, or pLNCX, pcDNA3.1(+/-), pcDNA/Zeo(+/-), pcDNA3.1/Hygro(+/-), PSVL, PMSG, pRSVcat, pSV2dhfr, pBC12MI and pCS2 etc. Commonly used mammalian cells include 293 cells, Cos1 cells, Cos7 cells, CV1 cells, mouse L cells and CHO cells, etc.
本文中引用的所有出版物，专利，和专利申请全部内容通过参考并入本文用于所有目的。All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.
具体实施方式detailed description
以下通过具体的实施例进一步说明本发明的技术方案，具体实施例不代表对本发明保护范围的限制。其他人根据本发明理念所做出的一些非本质的修改和调整仍属于本发明的保护范围。The technical solutions of the present invention are further described below through specific examples, which do not represent limitations to the protection scope of the present invention. Some non-essential modifications and adjustments made by others according to the concept of the present invention still belong to the protection scope of the present invention.
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.
实施例1抗体的制备方法The preparation method of embodiment 1 antibody
根据抗体的重链和轻链氨基酸序列设计重链和轻链的DNA序列，用PCR引物修饰DNA序列的5’和3’端，所述引物设计中分别包含重链和轻链的kozak序列及信号肽DNA序列。构建好的序列再克隆到现有重组抗体表达载体中，通过测序分析验证重组质粒的正确构建。将上述重组质粒转染表达细胞中进行表达，收集上清液、纯化获得抗体蛋白样品，并用于下面各种实施例。Design the DNA sequences of the heavy chain and light chain according to the amino acid sequences of the heavy chain and light chain of the antibody, and modify the 5' and 3' ends of the DNA sequence with PCR primers, which include the kozak sequences of the heavy chain and the light chain and the Signal peptide DNA sequence. The constructed sequence is then cloned into the existing recombinant antibody expression vector, and the correct construction of the recombinant plasmid is verified by sequencing analysis. The above-mentioned recombinant plasmids were transfected into expression cells for expression, and the supernatant was collected and purified to obtain antibody protein samples, which were used in various examples below.
其中，1)抗OX40抗体制备过程中，采用的表达载体为pCDNA3.1TM(+)(Invitrogen公司，货号为V79020)，抗体M-KF采用的表达细胞为α-(1,6)-岩藻糖基转移酶基因敲除的CHO细胞(岩藻糖基化水平约为0)，抗体M采用的表达细胞为CHO-S细胞(购自Invitrogen)；阳性对照抗体11D4(抗OX40抗体)的重链和轻链的序列来自专利US8236930B2，采用的表达载体为pCHO1.0质粒(购自Invitrogen)，表达细胞为CHO-S细胞(购自Invitrogen)；抗体M和M-KF的氨基酸序列见表1，抗体M和M-KF的核酸序列见表2，表2中框内为前导肽的序列；Among them, 1) In the preparation process of anti-OX40 antibody, the expression vector used is pCDNA3.1TM(+) (Invitrogen Company, product number is V79020), and the expression cell used for antibody M-KF is α-(1,6)-fucus Glycosyltransferase gene knockout CHO cells (fucosylation level is about 0), the expression cells used for antibody M are CHO-S cells (purchased from Invitrogen); positive control antibody 11D4 (anti-OX40 antibody) heavy The sequence of chain and light chain comes from patent US8236930B2, the expression vector used is pCHO1.0 plasmid (purchased from Invitrogen), and the expression cell is CHO-S cell (purchased from Invitrogen); the amino acid sequences of antibodies M and M-KF are shown in Table 1 , the nucleic acid sequences of antibodies M and M-KF are shown in Table 2, and the sequence of the leader peptide is in the box in Table 2;
2)抗TIGIT抗体制备过程中，采用的表达载体为pCDNA3.1 TM(+)，抗体h10D8OF采用的表达细胞为CHO细胞，抗体h10D8OFKF采用的表达细胞为α-(1,6)-岩藻糖基转移酶基因敲除的CHO细胞(岩藻糖基化水平约为0)；抗体h10D8OF和h10D8OFKF的氨基酸序列见表3； 2) During the preparation of the anti-TIGIT antibody, the expression vector used was pCDNA3.1 TM (+), the expression cell used for the antibody h10D8OF was CHO cells, and the expression cell used for the antibody h10D8OFKF was α-(1,6)-fucose CHO cells with base transferase gene knockout (fucosylation level is about 0); the amino acid sequences of antibodies h10D8OF and h10D8OFKF are shown in Table 3;
3)抗CTLA-4抗体准备过程中，采用的表达载体为pCDNA3.1 TM(+)，表达细胞为α-(1,6)-岩藻糖基转移酶基因敲除的CHO细胞(岩藻糖基化水平约为0)；抗CTLA-4抗体的氨基酸序列见表4； 3) During the preparation of anti-CTLA-4 antibody, the expression vector used was pCDNA3.1 TM (+), and the expression cell was α-(1,6)-fucosyltransferase gene knockout CHO cells (Fucus The glycosylation level is about 0); the amino acid sequence of the anti-CTLA-4 antibody is shown in Table 4;
4)抗PD-1抗体制备过程中，抗体A采用的表达载体为pCDNA3.1 TM(+)，表达细胞为CHO细胞；抗体A的氨基酸序列见表5。 4) During the preparation of the anti-PD-1 antibody, the expression vector used for antibody A is pCDNA3.1 TM (+), and the expression cell is CHO cells; the amino acid sequence of antibody A is shown in Table 5.
表1抗体M和M-KF的氨基酸序列Table 1 Amino acid sequences of antibodies M and M-KF
Figure PCTCN2022106326-appb-000005
Figure PCTCN2022106326-appb-000005
表2抗体M和M-KF的核酸序列The nucleotide sequence of table 2 antibody M and M-KF
Figure PCTCN2022106326-appb-000006
Figure PCTCN2022106326-appb-000006
Figure PCTCN2022106326-appb-000007
Figure PCTCN2022106326-appb-000007
表3抗体h10D8OF和h10D8OFKF的氨基酸序列Table 3 Amino acid sequences of antibodies h10D8OF and h10D8OFKF
Figure PCTCN2022106326-appb-000008
Figure PCTCN2022106326-appb-000008
Figure PCTCN2022106326-appb-000009
Figure PCTCN2022106326-appb-000009
表4抗CTLA-4抗体的氨基酸序列The amino acid sequence of table 4 anti-CTLA-4 antibody
Figure PCTCN2022106326-appb-000010
Figure PCTCN2022106326-appb-000010
表5抗PD-1抗体的氨基酸序列Table 5 Amino acid sequences of anti-PD-1 antibodies
Figure PCTCN2022106326-appb-000011
Figure PCTCN2022106326-appb-000011
实施例2体内药效试验Embodiment 2 drug efficacy test in vivo
1)肿瘤接种1) Tumor inoculation
在OX40人源化小鼠模型(购自百奥赛图)中研究本发明的抗体M-KF的抗肿瘤功效。将MC38肿瘤细胞以5×10 5个/0.1mL浓度接种于OX40人源化雌性小鼠的右侧皮下。 The anti-tumor efficacy of the antibody M-KF of the present invention was studied in the OX40 humanized mouse model (purchased from Biocytogen). MC38 tumor cells were inoculated subcutaneously on the right side of OX40 humanized female mice at a concentration of 5×10 5 cells/0.1 mL.
2)分组和给药2) Grouping and administration
待肿瘤生长到119mm 3时按肿瘤体积随机分组，每组6只。分组当天定义为D0天，并于D0天开始给药；给药日期为：第0天、第3天、第6天、第9天、第12天、第15天。抗体的给药途径为腹腔给药(i.p.)，按Q3D(每3天1次)的频率，三个剂量组1mg/kg、0.2mg/kg、0.04mg/kg，共给药6次。给药方案如表6所示。 When the tumor grew to 119mm 3 , they were randomly divided into groups according to the tumor volume, with 6 rats in each group. The grouping day was defined as D0 day, and the administration began on D0 day; the administration dates were: day 0, day 3, day 6, day 9, day 12, and day 15. The administration route of the antibody was intraperitoneal administration (ip), according to the frequency of Q3D (once every 3 days), three dosage groups of 1 mg/kg, 0.2 mg/kg, and 0.04 mg/kg were administered 6 times in total. The dosing regimen is shown in Table 6.
3)实验观察和数据采集3) Experimental observation and data collection
细胞接种后，每周常规监测肿瘤对动物正常行为的影响；具体指标包括小鼠的活动性，摄食和饮水情况，体重增加或降低情况，眼睛、被毛及其它异常情况。在整个研究期间每周测量两次肿瘤和体重，当肿瘤达到端点时或当小鼠具有20％体重减轻时使小鼠安乐死。肿瘤体积(mm 3)为0.5×(肿瘤长径×肿瘤短径 2)。 After cell inoculation, the effect of the tumor on the normal behavior of the animals was routinely monitored weekly; specific indicators included the mouse's activity, food intake and drinking conditions, weight gain or loss, eyes, coat and other abnormalities. Tumors and body weights were measured twice weekly throughout the study and mice were euthanized when tumors reached endpoint or when mice had 20% body weight loss. The tumor volume (mm 3 ) was 0.5×(tumor long diameter×tumor short diameter 2 ).
4)统计4) Statistics
各组小鼠的肿瘤体积、小鼠体重、肿瘤重量等实验结果以平均值±标准误差(mean±SEM)表示。数据采用SPSS进行分析。P<0.05为具有显著性差异。The experimental results such as tumor volume, mouse body weight, and tumor weight of mice in each group are expressed as mean ± standard error (mean ± SEM). Data were analyzed using SPSS. P<0.05 means significant difference.
TGItv(相对肿瘤体积的抑制率)计算公式：TGItv (inhibition rate relative to tumor volume) calculation formula:
TGItv＝(1-(mean RTV 给药组)/(mean RTV 对照组))×100％；mean RTV 给药组：给药组RTV平均值，mean RTV 对照组：对照组RTV平均值； TGItv=(1-(mean RTV administration group )/(mean RTV control group ))×100%; mean RTV administration group : RTV average value of administration group, mean RTV control group : control group RTV average value;
其中，RTV n＝V nt/V n0；V nt：编号为n的小鼠在第t天的肿瘤体积，V n0：编号为n的小鼠在第0天的肿瘤体积，RTV n：编号为n的小鼠在第t天的肿瘤相对体积。 Among them, RTV n =V nt /V n0 ; V nt : the tumor volume of the mouse numbered on day t, V n0 : the tumor volume of the mouse numbered on day 0, RTV n : the tumor volume of the mouse numbered The relative volume of the tumor in the mice of n at day t.
表6给药方案Table 6 Dosage regimen
Figure PCTCN2022106326-appb-000012
Figure PCTCN2022106326-appb-000012
备注：N为动物只数。Note: N is the number of animals.
如图1和表7所示，抗体M-KF的中、高剂量能明显抑制肿瘤的生长(P<0.05)，且效果明显好于抗体11D4的高剂量；各组之间的小鼠体重未发现明显差异，表明小鼠对抗体M-KF的药物耐受性良好。As shown in Figure 1 and Table 7, the medium and high doses of antibody M-KF can significantly inhibit the growth of tumors (P<0.05), and the effect is significantly better than the high dose of antibody 11D4; Significant differences were found, indicating that the mice tolerated the drug well against antibody M-KF.
表7第24天的肿瘤体积的抑制率Table 7 Inhibition rate of tumor volume on the 24th day
Figure PCTCN2022106326-appb-000013
Figure PCTCN2022106326-appb-000013
实施例3 B-hPD-1/hOX40双人源化小鼠的MC38结肠癌动物模型实验Example 3 MC38 colon cancer animal model experiment of B-hPD-1/hOX40 double humanized mice
将PBS重悬的MC38结肠癌细胞以5×10 5个/0.1mL浓度，0.1mL/只体积接种于B-hPD-1/hOX40人源化小鼠(北京百奥赛图基因生物技术有限公司)的右侧皮下。当平均肿瘤体积达到大约105mm 3时，挑选个体肿瘤体积合适的72只小鼠入组，将动物按肿瘤体积随机分配到9组，每组8只，分组当天开始给药，具体给药方案见下表8。 MC38 colon cancer cells resuspended in PBS were inoculated into B-hPD-1/hOX40 humanized mice at a concentration of 5 ×105/0.1mL, 0.1mL/vessel (Beijing Biocytogen Biotechnology Co., Ltd.) Subcutaneous on the right side. When the average tumor volume reached about 105 mm 3 , 72 mice with appropriate individual tumor volumes were selected to enter the group, and the animals were randomly assigned to 9 groups according to the tumor volume, with 8 mice in each group, and the administration began on the day of the grouping. For the specific dosage regimen, see Table 8 below.
药物评价指标有：肿瘤体积抑制率(TGItv)和瘤重抑制率(TGItw)：Drug evaluation indicators include: tumor volume inhibition rate (TGItv) and tumor weight inhibition rate (TGItw):
TGItv(％)＝[1-(Ti-T0)/(Vi-V0)]×100％(Ti：给药组在给药第i天的肿瘤体积均值，T0：治疗组在给药第0天的肿瘤体积均值；Vi：阴性对照组在给药第i天的肿瘤体积均值，V0：阴性对照组在给药第0天的肿瘤体积均值)；TGItv (%)=[1-(Ti-T0)/(Vi-V0)]×100% (Ti: the mean value of the tumor volume of the treatment group on the i-th day of administration, T0: the treatment group on the 0th day of administration The mean value of the tumor volume of the negative control group; Vi: the mean value of the tumor volume of the negative control group on the i-th day of administration, V0: the mean value of the tumor volume of the negative control group on the day 0 of the administration);
TGItw(％)＝(W阴性对照组-W治疗组)/W阴性对照组×100％，W指肿瘤重量。TGItw (%)=(W negative control group−W treatment group)/W negative control group×100%, W refers to tumor weight.
表8给药方案Table 8 Dosage regimen
Figure PCTCN2022106326-appb-000014
Figure PCTCN2022106326-appb-000014
1)小鼠在给药期间活动和进食状态良好，体重均有一定程度的上升，表明小鼠对受试药物的耐受性良好1) The mice had good activity and eating status during the administration period, and their body weight increased to a certain extent, indicating that the mice had good tolerance to the test drug
2)如表9所示，抗体M或M-KF与抗体A联合给药后，可以显著抑制肿瘤体积增长。2) As shown in Table 9, the combined administration of antibody M or M-KF and antibody A can significantly inhibit the growth of tumor volume.
表9第27天的肿瘤体积抑制率Table 9 Tumor volume inhibition rate on the 27th day
Figure PCTCN2022106326-appb-000015
Figure PCTCN2022106326-appb-000015
3)如表10所示，抗体M或M-KF与抗体A联合给药后，可以显著抑制肿瘤重量的增长。3) As shown in Table 10, the combined administration of antibody M or M-KF and antibody A can significantly inhibit the growth of tumor weight.
表10第27天的瘤重抑制率Table 10 The tumor weight inhibition rate on the 27th day
Figure PCTCN2022106326-appb-000016
Figure PCTCN2022106326-appb-000016
实施例4体外溶血实验和组织交叉反应试验Example 4 In vitro hemolysis test and tissue cross-reaction test
采用人血红细胞作为试验模型，当抗体M-KF注射液(浓度为25.8mg/mL)稀释超过10倍时，在体外不会引起人红细胞的溶血或凝集。Using human red blood cells as the test model, when the antibody M-KF injection (concentration of 25.8mg/mL) is diluted more than 10 times, it will not cause hemolysis or agglutination of human red blood cells in vitro.
采用生物素标记的抗体M-KF和链霉菌抗生物素蛋白-过氧化酶(SP)的免疫组化法，5和40μg/mL的抗体M-KF不与人和食蟹猴的组织发生组织交叉反应。Immunohistochemistry using biotin-labeled antibody M-KF and streptavidin-peroxidase (SP), 5 and 40 μg/mL antibody M-KF does not cross tissue from human and cynomolgus monkey tissues reaction.
实施例5抗体M-KF的临床研究Example 5 Clinical Research of Antibody M-KF
本研究是一项评价抗体M-KF单一给药以及抗体M-KF和抗PD-1抗体A联合给药在晚期恶性实体肿瘤患者中的安全性、耐受性、药代动力学(PK)特征和初步临床有效性的多中心、开放性、剂量递增的I期临床试验；探索最大耐受剂量(MTD)或最大给药剂量(MAD)，并为II期或后续临床研究提供推荐剂量及合理的给药方案。This study is an evaluation of the safety, tolerability, and pharmacokinetics (PK) of antibody M-KF single administration and antibody M-KF combined with anti-PD-1 antibody A in patients with advanced malignant solid tumors. Multi-center, open-label, dose-escalation phase I clinical trial of characteristics and preliminary clinical effectiveness; explore the maximum tolerated dose (MTD) or maximum administered dose (MAD), and provide recommended doses for phase II or follow-up clinical studies and Reasonable dosing regimen.
抗体M-KF通过静脉输液方式进行给药，输液的时间为60±5min，给药后的21天为DLT观察期；抗体M-KF的剂量为0.1mg/kg、0.3mg/kg、1mg/kg、3mg/kg、6mg/kg和10mg/kg每3周1次。首次给药抗体M-KF后，抗体A以300mg的剂量每3周一次进行静脉输液给药，输液的时间为60±5min。若每个剂量水平的抗体M-KF单药治疗的第一个周期(疗程)中没有发生DLT，将从第2周期开始进行300mg抗体A和抗体M-KF的组合治疗。Antibody M-KF is administered through intravenous infusion, the infusion time is 60±5min, and 21 days after administration is the DLT observation period; the dose of antibody M-KF is 0.1mg/kg, 0.3mg/kg, 1mg/kg kg, 3mg/kg, 6mg/kg and 10mg/kg every 3 weeks. After the first administration of antibody M-KF, antibody A was administered by intravenous infusion at a dose of 300 mg every 3 weeks, and the infusion time was 60±5 minutes. If DLT does not occur in the first cycle (course) of antibody M-KF monotherapy at each dose level, the combination therapy of 300 mg antibody A and antibody M-KF will start from the second cycle.
抗体A在注射抗体M-KF后以每3周1次的间隔进行给药。单例受试者将以0.1mg/kg抗体M-KF的起始剂量水平入组。若依据CTCAE v5.0 DLT观察期内没有发生可能与研究药物相关的2级或更高的毒性，则单例受试者的剂量递增至0.3mg/kg；若DLT观察期内单例受试者发生与研究药物相关的2级或更高的毒性，则将在此剂量水平招募另外2名受试者。在抗体M-KF单药治疗以及联合治疗中，DLT的观察期为3周。抗体M-KF从1mg/kg剂量开始采用基于“3+3”的剂量递增规则来探索安全剂量范围；抗体M-KF的剂量将递增到10mg/kg直到达到MTD或MAD。Antibody A was administered at intervals of once every 3 weeks after the injection of antibody M-KF. A single subject will be enrolled at a starting dose level of 0.1 mg/kg antibody M-KF. If there is no grade 2 or higher toxicity that may be related to the study drug during the DLT observation period according to CTCAE v5.0, the dose of the single subject will be increased to 0.3 mg/kg; if a single subject is tested during the DLT observation period If any grade 2 or higher toxicity related to the study drug occurs, an additional 2 subjects will be enrolled at this dose level. In antibody M-KF monotherapy and combination therapy, the observation period of DLT was 3 weeks. Antibody M-KF starts with a dose of 1 mg/kg and adopts a dose-escalation rule based on "3+3" to explore a safe dose range; the dose of antibody M-KF will be increased to 10 mg/kg until reaching the MTD or MAD.
MTD定义为在研究治疗的第一个周期(抗体M-KF第一次给药后21天)期间，6名受试者中不超过1名发生剂量限制性毒性(DLT)的最高剂量水平。剂量递增取决于治疗第一周期和第二周期期间观察到的毒性。The MTD was defined as the highest dose level at which no more than 1 of 6 subjects experienced a dose-limiting toxicity (DLT) during the first cycle of study treatment (21 days after the first dose of antibody M-KF). Dose escalation was based on observed toxicities during the first and second cycles of treatment.
受试者持续治疗直到出现：疾病进展、或不可耐受的毒性、或研究者的决定、或撤回知情同意、或研究终止，以先出现的为准。最长治疗时间为1年。在没有疾病进展的情况下继续接受治疗超过1年的受试者可以继续接受为期2年以上的下一个周期的药物治疗。如果患者出现输液相关反应并能够继续治疗，可基于临床实际情况可以使用苯海拉明或对乙酰氨基酚等进行预防给药。Subjects continued treatment until: disease progression, or unacceptable toxicity, or investigator's decision, or withdrawal of informed consent, or study termination, whichever occurred first. The maximum duration of treatment is 1 year. Subjects who continue to receive treatment for more than 1 year without disease progression can continue to receive the next cycle of drug treatment for more than 2 years. If the patient has an infusion-related reaction and can continue the treatment, diphenhydramine or acetaminophen can be used for prophylaxis based on the actual clinical situation.
安全性评价指标涉及：生命体征与体格检查、实验室检查(血常规、血生化、甲状腺功能、凝血常规、尿常规、便常规、妊娠试验)、心电图、不良事件(包括免疫相关不良事件)等。Safety evaluation indicators involve: vital signs and physical examination, laboratory examination (blood routine, blood biochemistry, thyroid function, coagulation routine, urine routine, stool routine, pregnancy test), electrocardiogram, adverse events (including immune-related adverse events), etc. .
所有剂量组的受试者在治疗期间(前6个治疗周期)的规定时间点需要收集血样，检测血清药物的浓度水平，研究药代动力学(PK)特征。PK涉及的参数有：C max、AUC (0-t)、AUC (0-∞)等。 Subjects in all dose groups need to collect blood samples at specified time points during the treatment period (the first 6 treatment cycles), detect the concentration level of the serum drug, and study the pharmacokinetic (PK) characteristics. The parameters involved in PK are: C max , AUC (0-t) , AUC (0-∞) and so on.
临床有效性评价：最佳总疗效(Best overall response)，客观缓解率(Objective Response Rate，ORR)，缓解持续时间(Duration of Response，DOR)，疾病控制率(Disease control rate，DCR)，无进展生存期(Progression-Free Survival，PFS)，总生存期(Overall survival，OS)。Clinical effectiveness evaluation: best overall response (Best overall response), objective response rate (Objective Response Rate, ORR), duration of response (Duration of Response, DOR), disease control rate (Disease control rate, DCR), no progression Survival (Progression-Free Survival, PFS), overall survival (Overall survival, OS).
AE将使用CTCAE v5.0进行分级。DLT被定义为在DLT观察期(抗体M-KF单药治疗的21天第一个周期，以及与抗体A联合治疗的21天的第二个周期)期间发生的毒性。与基础疾病、疾病进展、伴随药物或合并症明显相关除外，指定等级的所有AE都应计为DLT，且被认为至少可能与研究药物相关，具体如下：AEs will be graded using CTCAE v5.0. DLT was defined as the toxicity that occurred during the DLT observation period (21-day first cycle of antibody M-KF monotherapy, and 21-day second cycle of antibody A combination therapy). Unless clearly related to the underlying disease, disease progression, concomitant drugs, or comorbidities, all AEs of the assigned grade should be counted as DLTs and considered at least possibly related to the study drug, as follows:
5级毒性：Grade 5 Toxicity:
血液学毒性： Hematological Toxicity :
·4级血液学毒性(3-4级淋巴细胞减少症不认为是DLT)；·Grade 4 hematological toxicity (Grade 3-4 lymphopenia is not considered a DLT);
·4血小板减少症，持续时间≥7天；4 Thrombocytopenia, lasting ≥ 7 days;
·有出血倾向(≥2级出血)的或需要输注血小板的3级血小板减少症；·Thrombocytopenia with bleeding tendency (≥ grade 2 bleeding) or grade 3 thrombocytopenia requiring platelet transfusion;
·持续任何时间的4级中性粒细胞减少症，或持续时间≥7天或已存在感染的3级中性粒细胞减少症Grade 4 neutropenia lasting for any duration, or Grade 3 neutropenia lasting ≥ 7 days or pre-existing infection
·持续任何时间的3级或4级的中性粒细胞减少伴发热；·Grade 3 or 4 neutropenia with fever for any duration;
非血液学毒性： Non-hematological toxicity :
·3级非血液学毒性(非临床实验室)；· Grade 3 non-hematological toxicity (non-clinical laboratory);
·临床实验室检查发现的、满足以下条件之一的3、4级非血液学毒性：1)需要临床干预，2)异常并导致住院治疗的程度，3)导致第2周期治疗延迟≥2周的任何药物相关AE。Grade 3 and 4 non-hematological toxicity found in clinical laboratory tests that meet one of the following conditions: 1) requires clinical intervention, 2) is abnormal and leads to hospitalization, 3) leads to a delay of ≥ 2 weeks in the second cycle of treatment Any drug-related AEs.

Claims (23)

  1. 一种抗OX40抗体或抗原结合片段在制备与另一个治疗剂联合使用治疗肿瘤或癌症的药物的用途，所述抗OX40抗体或抗原结合片段包含SEQ ID NO:1所示的HCDR1、SEQ ID NO:2所示的HCDR2、SEQ ID NO:3所示的HCDR3、SEQ ID NO:4所示的LCDR1、SEQ ID NO:5所示的LCDR2和SEQ ID NO:6所示的LCDR3。An anti-OX40 antibody or antigen-binding fragment is used in the preparation of a drug for treating tumor or cancer in combination with another therapeutic agent, the anti-OX40 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO: 1, SEQ ID NO HCDR2 shown in :2, HCDR3 shown in SEQ ID NO:3, LCDR1 shown in SEQ ID NO:4, LCDR2 shown in SEQ ID NO:5, and LCDR3 shown in SEQ ID NO:6.
  2. 如权利要求1所述的用途，所述抗OX40抗体或抗原结合片段的重链可变区包含SEQ ID NO:7所示的序列，与SEQ ID NO:7所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:7所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或The use according to claim 1, the heavy chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 7, which has at least 80% compared with the sequence shown in SEQ ID NO: 7 A sequence of identity, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 7; and/or
    所述抗OX40抗体或抗原结合片段的轻链可变区包含SEQ ID NO:8所示的序列，与SEQ ID NO:8所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:8所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 8, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 8, or a sequence with SEQ ID NO: 8 Compared with the sequence shown in NO:8, there are one or more amino acid sequences with conservative amino acid substitutions.
  3. 如权利要求1所述的用途，所述抗OX40抗体或抗原结合片段的重链包含SEQ ID NO:9所示的序列，与SEQ ID NO:9所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:9所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或The use according to claim 1, the heavy chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 9, which has at least 80% identity compared with the sequence shown in SEQ ID NO: 9 sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:9; and/or
    所述抗OX40抗体或抗原结合片段的轻链包含SEQ ID NO:10所示的序列，与SEQ ID NO:10所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:10所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 10, a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO: 10, or a sequence with SEQ ID NO: 10 The sequences shown are compared to amino acid sequences having one or more conservative amino acid substitutions.
  4. 如权利要求1-3任一项所述的用途，所述另一个治疗剂选自针对以下靶点的抗体或抗原结合片段或抗体药物偶联物：EGFR、VEGF、VEGFR2、CTLA-4、PD-L1、PD-1、HER2、CD20、TROP2、LAG3、TIGIT、CD27、ICOS、BTLA、TIM3、BCMA、c-MET、TAA、CD19和4-1BB。The use according to any one of claims 1-3, wherein the other therapeutic agent is selected from antibodies or antigen-binding fragments or antibody-drug conjugates against the following targets: EGFR, VEGF, VEGFR2, CTLA-4, PD -L1, PD-1, HER2, CD20, TROP2, LAG3, TIGIT, CD27, ICOS, BTLA, TIM3, BCMA, c-MET, TAA, CD19 and 4-1BB.
  5. 如权利要求1-4任一项所述的用途，所述另一个治疗剂为抗PD-1抗体或抗原结合片段或抗PD-L1抗体或抗原结合片段。The use according to any one of claims 1-4, wherein the other therapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment or an anti-PD-L1 antibody or an antigen-binding fragment.
  6. 如权利要求1-4任一项所述的用途，所述另一个治疗剂为纳武利尤单抗、帕博利珠单抗、卡瑞利珠单抗、信迪利单抗、特瑞普利单抗或替雷利珠单抗。The use according to any one of claims 1-4, the other therapeutic agent is nivolumab, pembrolizumab, camrelizumab, sintilimab, toripril monoclonal antibody or tislelizumab.
  7. 如权利要求1-4任一项所述的用途，所述另一个治疗剂为抗PD-1抗体或抗原结合片段，其包含SEQ ID NO:25所示的HCDR1、SEQ ID NO:26所示的HCDR2、SEQ ID NO:27所示的HCDR3、SEQ ID NO:28所示的LCDR1、SEQ ID NO:29所示的LCDR2 和SEQ ID NO:30所示的LCDR3。The use according to any one of claims 1-4, the other therapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment, which comprises HCDR1 shown in SEQ ID NO:25, shown in SEQ ID NO:26 HCDR2 shown in SEQ ID NO:27, LCDR1 shown in SEQ ID NO:28, LCDR2 shown in SEQ ID NO:29, and LCDR3 shown in SEQ ID NO:30.
  8. 如权利要求7所述的用途，所述抗PD-1抗体或抗原结合片段的重链可变区包含SEQ ID NO:31所示的序列，与SEQ ID NO:31所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:31所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或The use according to claim 7, the heavy chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 31, compared with the sequence shown in SEQ ID NO: 31 has at least A sequence with 80% identity, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 31; and/or
    所述抗PD-1抗体或抗原结合片段的轻链可变区包含SEQ ID NO:32所示的序列，与SEQ ID NO:32所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:32所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 32, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 32, or a sequence with An amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:32.
  9. 如权利要求7所述的用途，所述抗PD-1抗体或抗原结合片段的重链包含SEQ ID NO:33所示的序列，与SEQ ID NO:33所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:33所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或The use according to claim 7, the heavy chain of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 33, which is at least 80% identical to the sequence shown in SEQ ID NO: 33 or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:33; and/or
    所述抗PD-1抗体或抗原结合片段的轻链包含SEQ ID NO:34所示的序列，与SEQ ID NO:34所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:34所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 34, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 34, or a sequence with SEQ ID NO Amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in :34.
  10. 如权利要求1所述的用途，其中所述药物的单位量为一次给药的量含所述抗OX40抗体0.6mg至900mg。The use according to claim 1, wherein the unit dose of the drug is 0.6 mg to 900 mg of the anti-OX40 antibody for one administration.
  11. 如权利要求1所述的用途，其中所述抗OX40抗体的每次给药量为0.6mg至900mg；或者，所述抗OX40抗体的每次给药量为0.01-25mg/kg。The use according to claim 1, wherein the dosage of each anti-OX40 antibody is 0.6 mg to 900 mg; or, the dosage of each anti-OX40 antibody is 0.01-25 mg/kg.
  12. 如权利要求7所述的用途，其中所述抗PD-1抗体给药的有效量为50mg至600mg；或者，所述抗PD-1抗体给药的有效量为1-10mg/kg。The use according to claim 7, wherein the effective dose of the anti-PD-1 antibody administered is 50 mg to 600 mg; or, the effective dose of the anti-PD-1 antibody administered is 1-10 mg/kg.
  13. 一种用于治疗肿瘤或癌症的方法，其包括：向有需要的患者给药有效量的抗OX40抗体或抗原结合片段和另一个治疗剂；A method for treating a tumor or cancer comprising: administering to a patient in need thereof an effective amount of an anti-OX40 antibody or antigen-binding fragment and another therapeutic agent;
    所述抗OX40抗体或抗原结合片段包含SEQ ID NO:1所示的HCDR1、SEQ ID NO:2所示的HCDR2、SEQ ID NO:3所示的HCDR3、SEQ ID NO:4所示的LCDR1、SEQ ID NO:5所示的LCDR2和SEQ ID NO:6所示的LCDR3。The anti-OX40 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, LCDR1 shown in SEQ ID NO:4, LCDR2 shown in SEQ ID NO:5 and LCDR3 shown in SEQ ID NO:6.
  14. 如权利要求13所述的方法，所述抗OX40抗体或抗原结合片段的重链可变区包含SEQ ID NO:7所示的序列，与SEQ ID NO:7所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:7所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列； 和/或The method of claim 13, wherein the heavy chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO:7, having at least 80% of the sequence shown in SEQ ID NO:7 A sequence of identity, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 7; and/or
    所述抗OX40抗体或抗原结合片段的轻链可变区包含SEQ ID NO:8所示的序列，与SEQ ID NO:8所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:8所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 8, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 8, or a sequence with SEQ ID NO: 8 Compared with the sequence shown in NO:8, there are one or more amino acid sequences with conservative amino acid substitutions.
  15. 如权利要求13所述的方法，所述抗OX40抗体或抗原结合片段的重链包含SEQ ID NO:9所示的序列，与SEQ ID NO:9所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:9所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或The method of claim 13, wherein the heavy chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO:9, which has at least 80% identity compared to the sequence shown in SEQ ID NO:9 sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:9; and/or
    所述抗OX40抗体或抗原结合片段的轻链包含SEQ ID NO:10所示的序列，与SEQ ID NO:10所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:10所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 10, a sequence having at least 80% identity compared to the sequence shown in SEQ ID NO: 10, or a sequence with SEQ ID NO: 10 The sequences shown are compared to amino acid sequences having one or more conservative amino acid substitutions.
  16. 如权利要求13-15任一项所述的方法，所述另一个治疗剂选自针对以下靶点的抗体或抗原结合片段或抗体药物偶联物：EGFR、VEGF、VEGFR2、CTLA-4、PD-L1、PD-1、HER2、CD20、TROP2、LAG3、TIGIT、CD27、ICOS、BTLA、TIM3、BCMA、c-MET、TAA、CD19和4-1BB。The method according to any one of claims 13-15, wherein the other therapeutic agent is selected from an antibody or an antigen-binding fragment or an antibody-drug conjugate against the following targets: EGFR, VEGF, VEGFR2, CTLA-4, PD -L1, PD-1, HER2, CD20, TROP2, LAG3, TIGIT, CD27, ICOS, BTLA, TIM3, BCMA, c-MET, TAA, CD19 and 4-1BB.
  17. 如权利要求13-16任一项所述的方法，所述另一个治疗剂为抗PD-1抗体或抗原结合片段或抗PD-L1抗体或抗原结合片段。The method of any one of claims 13-16, wherein the other therapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment or an anti-PD-L1 antibody or an antigen-binding fragment.
  18. 如权利要求13-16任一项所述的方法，所述另一个治疗剂为纳武利尤单抗、帕博利珠单抗、卡瑞利珠单抗、信迪利单抗、特瑞普利单抗或替雷利珠单抗；或者，The method according to any one of claims 13-16, the other therapeutic agent is nivolumab, pembrolizumab, camrelizumab, sintilimab, toripril monoclonal antibody or tislelizumab; or,
    所述另一个治疗剂为抗PD-1抗体或抗原结合片段，其包含SEQ ID NO:25所示的HCDR1、SEQ ID NO:26所示的HCDR2、SEQ ID NO:27所示的HCDR3、SEQ ID NO:28所示的LCDR1、SEQ ID NO:29所示的LCDR2和SEQ ID NO:30所示的LCDR3。The other therapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment comprising HCDR1 shown in SEQ ID NO:25, HCDR2 shown in SEQ ID NO:26, HCDR3 shown in SEQ ID NO:27, SEQ ID NO: LCDR1 shown in ID NO:28, LCDR2 shown in SEQ ID NO:29, and LCDR3 shown in SEQ ID NO:30.
  19. 如权利要求18所述的方法，所述抗PD-1抗体或抗原结合片段的重链可变区包含SEQ ID NO:31所示的序列，与SEQ ID NO:31所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:31所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或The method of claim 18, the heavy chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 31, compared with the sequence shown in SEQ ID NO: 31 has at least A sequence with 80% identity, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 31; and/or
    所述抗PD-1抗体或抗原结合片段的轻链可变区包含SEQ ID NO:32所示的序列，与SEQ ID NO:32所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:32所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain variable region of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 32, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 32, or a sequence with An amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:32.
  20. 如权利要求18所述的方法，所述抗PD-1抗体或抗原结合片段的重链包含SEQ  ID NO:33所示的序列，与SEQ ID NO:33所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:33所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列；和/或The method of claim 18, the heavy chain of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO:33, which is at least 80% identical to the sequence shown in SEQ ID NO:33 or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:33; and/or
    所述抗PD-1抗体或抗原结合片段的轻链包含SEQ ID NO:34所示的序列，与SEQ ID NO:34所示序列相比具有至少80％同一性的序列，或与SEQ ID NO:34所示序列相比具有一个或多个保守氨基酸取代的氨基酸序列。The light chain of the anti-PD-1 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 34, a sequence having at least 80% identity compared with the sequence shown in SEQ ID NO: 34, or a sequence with SEQ ID NO Amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in :34.
  21. 如权利要求13-20任一项所述的方法，每个治疗周期内抗OX40抗体给药的有效量为0.6mg至900mg；或者，所述抗OX40抗体的每次给药量为0.01-25mg/kg。The method according to any one of claims 13-20, the effective dose of anti-OX40 antibody administered in each treatment cycle is 0.6 mg to 900 mg; or, the dose of each anti-OX40 antibody administered is 0.01-25 mg /kg.
  22. 如权利要求16-20任一项所述的方法，每个治疗周期内抗PD-1抗体给药的有效量为50mg至600mg；或者，所述抗PD-1抗体的每次给药量为1-10mg/kg。The method according to any one of claims 16-20, the effective dose of anti-PD-1 antibody administered in each treatment cycle is 50 mg to 600 mg; or, the dose of each anti-PD-1 antibody administered is 1-10mg/kg.
  23. 一种试剂盒，其包含抗OX40抗体或抗原结合片段、另一个治疗剂，以及用于指导有需要患者给药抗OX40抗体或抗原结合片段和另一个治疗剂的说明书；A kit comprising an anti-OX40 antibody or antigen-binding fragment, another therapeutic agent, and instructions for instructing a patient in need of administering the anti-OX40 antibody or antigen-binding fragment and another therapeutic agent;
    所述抗OX40抗体或抗原结合片段包含SEQ ID NO:1所示的HCDR1、SEQ ID NO:2所示的HCDR2、SEQ ID NO:3所示的HCDR3、SEQ ID NO:4所示的LCDR1、SEQ ID NO:5所示的LCDR2和SEQ ID NO:6所示的LCDR3。The anti-OX40 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, HCDR3 shown in SEQ ID NO:3, LCDR1 shown in SEQ ID NO:4, LCDR2 shown in SEQ ID NO:5 and LCDR3 shown in SEQ ID NO:6.
PCT/CN2022/106326 2021-07-19 2022-07-18 Application of anti-ox40 antibody in combined drug WO2023001118A1 (en)

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