WO2022105839A1 - 抗ox40抗体在治疗肿瘤或癌症中的应用 - Google Patents

抗ox40抗体在治疗肿瘤或癌症中的应用 Download PDF

Info

Publication number
WO2022105839A1
WO2022105839A1 PCT/CN2021/131542 CN2021131542W WO2022105839A1 WO 2022105839 A1 WO2022105839 A1 WO 2022105839A1 CN 2021131542 W CN2021131542 W CN 2021131542W WO 2022105839 A1 WO2022105839 A1 WO 2022105839A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
seq
sequence
weeks
amino acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2021/131542
Other languages
English (en)
French (fr)
Inventor
梁世忠
梁炳辉
俞金泉
李骁
宋述强
李胜峰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Bio Thera Solutions Ltd
Original Assignee
Bio Thera Solutions Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Bio Thera Solutions Ltd filed Critical Bio Thera Solutions Ltd
Priority to CN202180073819.9A priority Critical patent/CN116507365A/zh
Priority to US18/037,401 priority patent/US20230416386A1/en
Priority to EP21893986.6A priority patent/EP4248995A4/en
Publication of WO2022105839A1 publication Critical patent/WO2022105839A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2878—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
    • C07K2317/41—Glycosylation, sialylation, or fucosylation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/51—Complete heavy chain or Fd fragment, i.e. VH + CH1
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/515—Complete light chain, i.e. VL + CL
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

Definitions

  • the invention belongs to the field of biomedicine, and particularly relates to the application of anti-OX40 antibody in the treatment of tumor or cancer.
  • 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 the T cell antigen receptors recognizing the antigen, and the activation signal is transferred into the cell through the CD3 molecule; while the second signal is called the costimulatory signal, which is composed of the costimulatory molecules on the surface of the antigen-presenting cells or target cells and the activated Produced by interaction with co-stimulatory molecule receptors on the surface of T cells.
  • Costimulatory signals promote antigen-specific T cell proliferation and differentiation 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 antigen, OX40 molecules will be highly expressed within 1-3 days. Activation of OX40 signaling further enhances T cell activation signaling to enhance the immune system response (Gramaglia I et al. J. Immunol. 2000; 165:3043-3050).
  • anti-OX40 antibodies activate the immune system and inhibit tumors mainly through the following three cellular physiological mechanisms.
  • the third is to deplete Treg cells and reduce their inhibition of effector T cells through ADCC or ADCP, etc.
  • OX40 is a very potential activating target in tumor immunotherapy, which can provide a new means for tumor immunotherapy.
  • the present invention discloses methods or uses of anti-OX40 antibodies or antigen-binding fragments for treating tumors or cancers.
  • the anti-OX40 antibody or antigen-binding fragment agent is used to treat tumors or cancer.
  • the anti-OX40 antibody or antigen-binding fragment comprises at least HCDR1 set forth in SEQ ID NO:1, HCDR2 set forth in SEQ ID NO:2, HCDR3 set forth in SEQ ID NO:3, : one or more of LCDR1 shown in SEQ ID NO: 5, LCDR2 shown in SEQ ID NO: 6, and LCDR3 shown in SEQ ID NO: 6.
  • the anti-OX40 antibody or antigen-binding fragment comprises HCDR1 set forth in SEQ ID NO:1, HCDR2 set forth in SEQ ID NO:2, HCDR3 set forth 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, or a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO:7 , 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
  • the light chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence shown in SEQ ID NO: 8, or a sequence with at least 80% identity to the sequence shown in SEQ ID NO: 8, or a sequence with SEQ ID NO: 8 :
  • the sequence shown in 8 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
  • the heavy chain variable region of the anti-OX40 antibody or antigen-binding fragment comprises the sequence set forth 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.
  • the heavy chain of the anti-OX40 antibody comprises the sequence set forth in SEQ ID NO:9, or a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO:9, or the sequence set forth in SEQ ID NO:9 The amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in 9; and/or
  • the light chain of the anti-OX40 antibody comprises the sequence shown in SEQ ID NO:10, or a sequence with at least 80% identity to the sequence shown in SEQ ID NO:10, or compared with the sequence shown in SEQ ID NO:10 An amino acid sequence with 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 Contains the sequence shown in SEQ ID NO: 10
  • antibody M or M-KF contains two heavy chains with the same sequence and two light chains with the same sequence.
  • the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment has a fucosylation level of 0-10%. In some embodiments, the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment has a fucosylation level of 0-5%.
  • the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment has a level of fucosylation of 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 A range between values (including the endpoints) or any value within it.
  • the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment does not bind fucose.
  • the anti-OX40 antibody (eg, antibody M-KF) or antigen-binding fragment has an enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
  • Anti-OX40 antibodies or antigen-binding fragments can be expressed in CHO cells or HEK293 cells by genetic engineering and obtained by purification; purification can be carried out by conventional methods, such as centrifuging the cell suspension and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as Protein A affinity columns and ion exchange columns can be used to purify antibody proteins.
  • the anti-OX40 antibody or antigen-binding fragment is expressed by an ⁇ -(1,6)-fucosyltransferase knockout cell line, such as CHO-BAT-KF cells disclosed in PCT/CN2018/100008 Tie.
  • the antibody M-KF is expressed by the CHO-BAT-KF cell line.
  • the method or use comprises administering to a patient in need thereof an effective amount of an anti-OX40 antibody or antigen-binding fragment.
  • the anti-OX40 antibody is antibody M-KF.
  • the antibody M-KF is expressed by an a-(1,6)-fucosyltransferase knockout cell line.
  • the anti-OX40 antibody is administered in an effective dose of about 0.6 mg to 900 mg per dose.
  • 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, leukemia, lymphoma, and myeloma.
  • the leukemia includes acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and myeloproliferative disorders/neoplastics (MPDS) ).
  • ALL acute lymphocytic leukemia
  • AML acute myeloid leukemia
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • MPDS myeloproliferative disorders/neoplastics
  • lymphomas include Hodgkin's lymphoma, indolent and aggressive non-Hodgkin's lymphoma, Burkitt's lymphoma, and follicular lymphoma (small cell and large cell).
  • the myeloma includes multiple myeloma (MM), giant cell myeloma, heavy chain myeloma, and light chain or Bence-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 tumor and cancer are pathologically confirmed locally advanced or metastatic malignant solid tumors for which there is no effective treatment.
  • 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 or antigen-binding fragment, wherein the administration of the anti-OX40 antibody or antigen-binding fragment is effective
  • the amount is about 0.6 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 of its values.
  • the anti-OX40 antibody is antibody M-KF.
  • the antibody M-KF is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line.
  • an anti-OX40 antibody can be formulated into a pharmaceutical composition and administered to a patient in a variety of forms suitable for the chosen route of administration, eg, parenteral, intravenous (iv), intramuscular , topical or subcutaneous.
  • the anti-OX40 antibody can be infused intravenously.
  • the amount of anti-OX40 administered will depend on the nature of the drug, the extent to which the cell surface triggers internalization, transport, and release of the drug, as well as 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 administered at about 0.01 mg/kg to 25 mg/kg or formulation containing such dose of anti-OX40 antibody per administration.
  • 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 0.2 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg, about 0.9 mg/kg, about 1 mg/kg, about 2 mg/kg, about 2.5 mg/kg, about 3 mg/kg, about 4 mg/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 17
  • the present invention discloses a method for treating a tumor or cancer, the method comprising: administering to a patient in need thereof about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg every 3 weeks /kg, about 0.06 mg/kg, about 0.08 mg/kg, about 0.1 mg/kg, about 0.2 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg, about 0.9 mg/kg, about 1 mg/kg, About 2 mg/kg, about 2.5 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, about 10 mg/kg , about 11 mg/kg, about 12 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15 mg/kg, about 16 mg/kg, about 17 mg/kg, about 18 mg/kg, about 19 mg/kg, about 20 mg/kg , about 21 mg/kg, about 22 mg/kg, about 23 mg/kg,
  • the present invention discloses a method for treating a tumor or cancer, the method comprising: administering to a patient in need thereof about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg every 3 weeks /kg, about 0.06 mg/kg, about 0.08 mg/kg, about 0.1 mg/kg, about 0.2 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg, about 0.9 mg/kg, about 1 mg/kg, About 2 mg/kg, about 2.5 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, about 10 mg/kg , about 11 mg/kg, about 12 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15 mg/kg, about 16 mg/kg, about 17 mg/kg, about 18 mg/kg, about 19 mg/kg, about 20 mg/kg , about 21 mg/kg, about 22 mg/kg, about 23 mg/kg,
  • the present invention discloses a method for treating a tumor or cancer, the method comprising: administering to a patient in need thereof about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg every 3 weeks /kg, about 0.06 mg/kg, about 0.08 mg/kg, about 0.1 mg/kg, about 0.2 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg, about 0.9 mg/kg, about 1 mg/kg, About 2 mg/kg, about 2.5 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, about 10 mg/kg , about 11 mg/kg, about 12 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15 mg/kg, about 16 mg/kg, about 17 mg/kg, about 18 mg/kg, about 19 mg/kg, about 20 mg/kg , about 21 mg/kg, about 22 mg/kg, about 23 mg/kg,
  • the present invention discloses a method of treating a tumor or cancer, comprising administering an effective amount of an anti-OX40 antibody (or preparation) to a patient in need thereof; wherein the effective amount of the anti-OX40 antibody is a single administration
  • the drug is about 0.6 mg to 900 mg (or a preparation containing this dose of anti-OX40 antibody). Dosage schedule and mode of administration depend on the benefit-risk assessment of the anti-OX40 antibody (or formulation) in certain patient populations and general clinical practice guidelines.
  • the effective amount of anti-OX40 antibody administered to a patient per treatment cycle is about 0.6 mg to 900 mg (or a formulation containing such dose of anti-OX40 antibody).
  • the effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) administered to a patient per treatment cycle is about 0.6 mg, about 1 mg, about 1.8 mg, about 6 mg, about 10 mg, about 18 mg, about 30 mg, about 60 mg, about 100 mg, about 120 mg, about 180 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, about 700 mg, about 800 mg, about 900 mg, or a range between any two of these values (including endpoints) or any value therein, or a formulation containing such dose of anti-OX40 antibody.
  • an anti-OX40 antibody eg, antibody M or M-KF
  • one treatment cycle is administered once every 1 week to 7 weeks.
  • the effective amount of anti-OX40 antibody administered in each treatment cycle is about 0.6 mg to 600 mg, or a formulation containing such dose of anti-OX40 antibody; wherein one 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.
  • 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 the anti-OX40 antibody in each treatment cycle from about 0.6 mg to about 6 mg, or a formulation containing such dose of the 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 0.6 mg, about 1 mg, about 1.5 mg, about 1.8 mg, about 2 mg, about 2.9 mg, about 3.6 mg, about 4 mg, about 4.3 mg, about 5 mg, about 5.5 mg, about 6 mg, or a range between any two of these values (including endpoints) or any value therein, or a formulation containing this dose of anti-OX40 antibody; wherein, one treatment cycle For 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 60 mg to 600 mg, or a formulation containing such dose of the 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 60 mg, about 80 mg, about 90 mg, about 100 mg, about 120 mg, about 160 mg, about 180 mg, about 200 mg, about 243 mg, about 290 mg, about 355 mg, about 400 mg, about 425 mg, about 480 mg, about 500 mg, about 565 mg, about 600 mg, or a range between any two of these values (including endpoints) or any value therein, or 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 600 mg to 900 mg, or a formulation containing such dose of the 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 600 mg, about 680 mg, about 700 mg, about 750 mg, about 780 mg, about 800 mg, about 820 mg, about 900 mg, or any of these values
  • an effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) administered to a patient per treatment cycle is about 0.5 mg to 1 mg, or a formulation containing such an amount of anti-OX40 antibody; such as about 0.6 mg administered 1 medicine.
  • the effective amount of the anti-OX40 antibody administered to the patient per treatment cycle is about 0.6 mg, or a formulation containing this dose of the anti-OX40 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks weeks or about 4 weeks.
  • the patient is administered an effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) per treatment cycle of about 1.5 mg to 2.2 mg, or a formulation containing such an amount of anti-OX40 antibody; such as about 1.8 mg Administer 1 time.
  • the effective amount of the anti-OX40 antibody administered to the patient per treatment cycle is about 1.8 mg, or a formulation containing this dose of the anti-OX40 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks weeks or about 4 weeks.
  • an effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) administered to a patient per treatment cycle is about 4 mg to 8 mg, or a formulation containing such an amount of anti-OX40 antibody; such as about 6 mg administered 1 Second-rate.
  • the patient is administered an effective amount of anti-OX40 antibody per treatment cycle of about 6 mg, or a formulation containing such dose of anti-OX40 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
  • an effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) administered to a patient per treatment cycle is about 50 mg to 76 mg, or a formulation containing such an amount of anti-OX40 antibody; such as about 60 mg administered1 Second-rate.
  • the effective amount of anti-OX40 antibody administered to the patient per treatment cycle is about 60 mg, or a formulation containing such 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) per treatment cycle of about 167 mg to 189 mg, or a formulation containing such an amount of anti-OX40 antibody; such as about 180 mg administered 1 Second-rate.
  • an anti-OX40 antibody eg, antibody M or M-KF
  • the effective amount of anti-OX40 antibody administered to the patient per treatment cycle is about 180 mg, or a formulation containing such 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) per treatment cycle of about 390 mg to 410 mg, or a formulation containing such an amount of anti-OX40 antibody; such as about 400 mg administered 1 Second-rate.
  • an anti-OX40 antibody eg, antibody M or M-KF
  • the effective amount of the anti-OX40 antibody administered to the patient per treatment cycle is about 400 mg, or a formulation containing such dose of the 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) per treatment cycle of about 493 mg to 512 mg, or a formulation containing this dose of an anti-OX40 antibody; such as about 500 mg administered 1 Second-rate.
  • an anti-OX40 antibody eg, antibody M or M-KF
  • the effective amount of anti-OX40 antibody administered to the patient per treatment cycle is about 500 mg, or a formulation containing such dose of anti-OX40 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
  • an effective amount of an anti-OX40 antibody (eg, antibody M or M-KF) administered to a patient per treatment cycle is about 588 mg to 606 mg, or a formulation containing such dose of an anti-OX40 antibody; such as about 600 mg administered 1 Second-rate.
  • the effective amount of anti-OX40 antibody administered to the patient per treatment cycle is about 600 mg, or a formulation containing such 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) per treatment cycle of about 882 mg to 916 mg, or a formulation containing such an amount of anti-OX40 antibody; such as about 900 mg administered 1 Second-rate.
  • an anti-OX40 antibody eg, antibody M or M-KF
  • the effective amount of the anti-OX40 antibody administered to the patient per treatment cycle is about 900 mg, or a formulation containing such dose of the anti-OX40 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks or about 4 weeks.
  • the anti-OX40 antibody is administered in an effective amount of about 0.6 mg to 600 mg once every 3 weeks. In some embodiments, the anti-OX40 antibody is administered in an effective amount of 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 3 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 18 mg, about 60 mg, or about 180 mg every 3 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 600 mg to 900 mg once every 3 weeks. In some embodiments, the effective amount of anti-OX40 antibody administered is about 700 mg, about 800 mg, or about 900 mg every 3 weeks.
  • the patient is administered an anti-OX40 antibody (or formulation) once per treatment cycle.
  • the anti-OX40 antibody (or formulation) is administered multiple times per treatment cycle, eg, 2, 3, 4, or 5 times.
  • the patient is only dosed 1 or 4 times per treatment cycle.
  • the patient is treated with one treatment cycle. In some embodiments, the patient is treated with multiple (eg, 2, 3, or 4) treatment cycles. In some embodiments, the patient receives treatment until the condition resolves and no longer requires treatment.
  • the present invention discloses a method for treating a tumor or cancer, the method comprising: administering to a patient in need thereof about 0.6 mg to 6 mg, about 6 mg to 60 mg, about 60 mg to 180 mg every 3 weeks , about 180 mg to 600 mg, about 600 mg to 900 mg, such as about 0.6 mg, about 1.8 mg, about 6 mg, about 18 mg, about 60 mg, 100 mg, about 180 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, or about 900 mg of anti-OX40 antibody, or a preparation containing this dose of anti-OX40 antibody.
  • the anti-OX40 antibody is Antibody M.
  • the anti-OX40 antibody is antibody M-KF.
  • about 0.6 mg of the anti-OX40 antibody is administered once every 3 weeks. In some embodiments, about 1.8 mg of the anti-OX40 antibody is administered once every 3 weeks. In some embodiments, about 6 mg of the anti-OX40 antibody is administered once every 3 weeks. In some embodiments, about 18 mg of the anti-OX40 antibody is administered once every 3 weeks. In some embodiments, about 60 mg of the anti-OX40 antibody is administered once every 3 weeks. In some embodiments, about 180 mg of the anti-OX40 antibody is administered once every 3 weeks. In some embodiments, about 600 mg of the anti-OX40 antibody is administered once every 3 weeks. In some embodiments, about 900 mg of the anti-OX40 antibody is administered once every 3 weeks.
  • the patient's symptoms are relieved after a single dose is administered. In some embodiments, after a single dose is administered and the patient's symptoms are not relieved as expected, the patient is administered about 0.6 mg to 900 mg of the anti-OX40 antibody until the patient's symptoms are relieved.
  • the anti-OX40 antibody (or formulation) is administered by subcutaneous (s.c.) injection, intraperitoneal (i.p.) injection, parenteral injection, intraarterial injection, or intravenous (i.v.) injection.
  • the anti-OX40 antibody (or formulation) is administered by infusion.
  • the anti-OX40 antibody (or formulation) is administered as a bolus injection.
  • the anti-OX40 antibody (or formulation) is administered by intravenous (i.v.) infusion (ie, intravenous infusion).
  • intravenous infusion ie, intravenous 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 (including the endpoints) between any two of these values, or any value therein. In some embodiments, the duration of the intravenous infusion is > 60 minutes.
  • the anti-OX40 antibody (or formulation) is used in combination with other therapeutic methods to treat tumors or cancers, such as chemotherapy, radiotherapy, immunotherapy, hormone therapy, targeted therapy, biological therapy, surgery, and the like.
  • the anti-OX40 antibody (or formulation) is combined with other tumor or cancer therapeutic agents to treat tumors or cancer, such as hormones, antibodies to treat tumors or cancer, and the like.
  • the present invention discloses the application of the anti-OX40 antibody in the preparation of a medicament for the treatment of tumor or cancer.
  • the drug for treating a tumor or cancer includes an anti-OX40 antibody.
  • the anti-OX40 antibody is Antibody M.
  • the anti-OX40 antibody is antibody M-KF.
  • the antibody M-KF is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line.
  • the present invention also discloses a kit comprising an anti-OX40 antibody (or preparation) and instructions for instructing a patient in need to administer the anti-OX40 antibody (or preparation).
  • the anti-OX40 antibody is Antibody M.
  • the anti-OX40 antibody is antibody M-KF.
  • the antibody M-KF is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line.
  • the present invention also discloses a pharmaceutical composition suitable for injection comprising the anti-OX40 antibody, such as a bolus injection type pharmaceutical composition or an infusion (drip) type pharmaceutical composition.
  • Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (herein water-soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
  • suitable carriers include physiological saline, bacteriostatic or phosphate buffered saline (PBS), ethanol, solvents or dispersion media of polyols (eg, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and the like, and suitable mixture.
  • the pharmaceutical composition further includes a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier may be implemented comprising antibacterial and/or antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
  • the pharmaceutically acceptable carrier may contain isotonic agents such as sugars, polyols (such as mannitol, sorbitol), sodium chloride.
  • the pharmaceutical composition comprises at least 0.1% anti-OX40 antibody. The percentage of antibody can vary and is between about 2% and 90% by weight of a given dosage form. The amount of anti-OX40 antibody in such a therapeutically useful pharmaceutical composition can be an effective amount for administration.
  • the anti-OX40 antibody is Antibody M. In some embodiments, the anti-OX40 antibody is antibody M-KF. In some embodiments, the antibody M-KF is expressed by an alpha-(1,6)-fucosyltransferase knockout cell line.
  • the present invention also discloses a preparation method of the above-mentioned pharmaceutical composition: respectively mixing the anti-OX40 antibody described herein with a pharmaceutically acceptable carrier suitable for injection (eg, water for injection, physiological saline, etc.).
  • a pharmaceutically acceptable carrier suitable for injection eg, water for injection, physiological saline, etc.
  • Methods of admixing the above-mentioned anti-OX40 antibody with a pharmaceutically acceptable carrier are generally known in the art.
  • the anti-OX40 antibody (or preparation) of the present invention can be used for the treatment of tumor or cancer.
  • Figure 1 shows that the anti-OX40 antibody M-KF inhibits the proliferation of tumor cells; the ordinate represents the tumor volume.
  • an entity refers to one or more of such entities, eg "an antibody” should be understood to mean one or more antibodies, thus the term “an” (or “an” ), “one or more” and “at least one” are used interchangeably herein.
  • polypeptide is intended to encompass the singular “polypeptide” as well as the plural “polypeptide”, and refers to a molecule consisting of amino acid monomers linked linearly by amide bonds (also known as peptide bonds).
  • polypeptide refers to any single chain or chains of two or more amino acids, and does not refer to a particular length of the product.
  • the definition of “polypeptide” includes a peptide, dipeptide, tripeptide, oligopeptide, "protein”, “amino acid chain” or any other term used to refer to two or more amino acid chains, and the term “polypeptide” may Used in place of, or used interchangeably with, any of the above terms.
  • polypeptide is also intended to refer to the product 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-native Amino acid modifications that occur.
  • a polypeptide may be derived from a natural biological source or produced by recombinant techniques, but it need not be translated from a given nucleic acid sequence, and it may be produced by any means including chemical synthesis.
  • Amino acid refers to an organic compound containing both an amino group and a carboxyl group, such as an alpha-amino acid, which can be encoded by a nucleic acid directly or in a precursor form.
  • 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 same amino acid is encoded by different codons 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).
  • alanine three-letter code: ala, one-letter code: A
  • arginine arg, R
  • asparagine asparag
  • Constant amino acid substitution refers to the replacement of one amino acid residue by another amino acid residue containing a side chain (R group) of similar chemical properties (eg, charge or hydrophobicity). In general, conservative amino acid substitutions will not substantially alter the functional properties of the protein.
  • amino acid classes containing 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 for "conservative amino acid substitutions of VL and 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 (including endpoints) between any two of these values, or any value therein.
  • the number of amino acids for a "conservative amino acid substitution of a 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 (including endpoints) between any two of these values, or any value therein.
  • encoding when applied to a polynucleotide refers to a polynucleotide referred to as “encoding” a polypeptide, transcribed and/or in its native state or when manipulated by methods well known to those skilled in the art Or translation can yield the polypeptide and/or fragments thereof.
  • recombinant refers to a polypeptide or polynucleotide and means a form of the polypeptide or polynucleotide that does not occur in nature, non-limiting examples may be combined to produce polynucleotides that do not normally exist or peptide.
  • Identity refers to the sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing the positions within each sequence that can be aligned. A molecule is homologous when a position in the sequences being compared is occupied by the same base or amino acid. The degree of homology between sequences is a function of the number of matches 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 (including endpoints) between any two values in a numerical value or any value therein.
  • a polynucleotide or polynucleotide sequence has a certain percentage (eg, 90%, 95%, 98% or 99%) "identity" or “sequence identity” to 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.
  • the percent alignment 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. Alignments are preferably performed using default parameters.
  • Biologically equivalent polynucleotides are polynucleotides that have the above-specified percentages of identity and encode polypeptides having the same or similar biological activity.
  • Antibody refers to a polypeptide or polypeptide complex that specifically recognizes and binds 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 in the molecule that contains at least a portion of an immunoglobulin molecule that has the biological activity of binding to an antigen.
  • Antibodies and antigen-binding fragments include, but are not limited to, the 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), a light chain constant region (CL), a 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).
  • antibody includes a wide variety of biochemically distinguishable polypeptides. Those of skill in the art will appreciate that classes of heavy chains include gamma, mu, alpha, delta, or epsilon (gamma, mu, alpha, delta, epsilon), with some subclasses (eg, gamma1-gamma4). The nature of this chain determines the "class” of the antibody as IgG, IgM, IgA, IgG or IgE, respectively. Immunoglobulin subclasses (isotypes), eg, IgGl, IgG2, IgG3, IgG4, IgG5, etc., are well characterized and the functional specificities conferred are known.
  • the immunoglobulin molecule is of the IgG class.
  • the four chains are connected in a "Y" configuration by disulfide bonds, where the light chain begins at the "Y" mouth and continues through the variable region surrounding the heavy chain.
  • Antibodies, antigen-binding fragments or derivatives disclosed herein include, but are not limited to, polyclonal, monoclonal, multispecific, fully human, humanized, primatized, chimeric antibodies, single chain antibodies, epitope binding Fragments (eg, Fab-like, Fab'-like, and F(ab') 2 ), single-chain-like Fvs (scFv).
  • Light chains can be classified as kappa ( ⁇ ) or lambda ( ⁇ ). Each heavy chain can bind to a kappa or lambda light chain.
  • ⁇ kappa
  • ⁇ lambda
  • the amino acid sequence extends from the N-terminus of the forked terminus in the Y configuration to the C-terminus at the bottom of each chain.
  • the variable region of immunoglobulin kappa light chain is V ⁇ ; the variable region of immunoglobulin ⁇ light chain is V ⁇ .
  • variable regions of the light (VL) and heavy (VH) chain portions 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, and the like. By convention, the numbering of constant regions increases as they become further 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-terminus of the heavy and light chains, respectively.
  • CDR complementarity determining region
  • CDRs as defined by Kabat and Chothia include overlaps or subsets of amino acid residues when compared to each other. Nonetheless, it is within the scope of the invention to apply either definition to refer to the CDRs of an antibody or variant thereof.
  • the exact residue numbers encompassing a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in the art can usually determine which specific residues the CDRs contain based on the amino acid sequence of the variable region of the antibody.
  • Kabat et al. also define a numbering system applicable to variable region sequences of any antibody.
  • One of ordinary skill in the art can apply this "Kabat numbering" system to any variable region sequence independent of experimental data other than the sequence itself.
  • Kabat Numbering means 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 may also use the EU or Chothia numbering system.
  • Treatment means therapeutic treatment and prophylactic or prophylactic measures, the purpose of which is to prevent, slow, ameliorate and stop adverse physiological changes or disorders, such as the progression of disease, including but not limited to the following whether detectable or undetectable As a result, alleviation of symptoms, reduction of disease severity, stabilization of disease state (ie, no worsening), delay or slowdown of disease progression, improvement or alleviation of disease state, alleviation or disappearance (whether in part or in whole), prolongation and Expected duration of survival when not receiving treatment, etc.
  • a patient in need of treatment includes a patient already suffering from a condition or disorder, a patient susceptible to a condition or disorder, or a patient in need of prevention of such a condition or disorder, may or may be expected from administration of the antibodies or compositions 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.
  • Effective amount refers to the amount of 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 term "in need” means that a patient has been identified as in need of a particular method or treatment. In some embodiments, identification can be made by any diagnostic means. In any of the methods and treatments described herein, the patient may be in need of such methods and treatments.
  • Antibody-encoding DNA can be designed and synthesized according to the antibody amino acid sequences described herein by conventional methods, inserted into an expression vector, and then transfected into host cells, and the transfected host cells are cultured in culture to produce monoclonal antibodies.
  • 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.
  • Efficient transcription can be obtained by the early and late promoters of SV40, long terminal repeats from retroviruses such as RSV, HTLV1, HIVI, and the early promoter of cytomegalovirus, and other cellular promoters such as muscle can be used.
  • 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.
  • the DNA sequences of the light and heavy chains were constructed from the heavy and light chain amino acid sequences of the antibody.
  • the 5' and 3' ends of the DNA sequence were modified with PCR primers designed to add appropriate leader sequences to each chain, and then cloned into the existing recombinant antibody expression vector, and the correct construction of the recombinant plasmid was verified by sequencing analysis.
  • the above recombinant plasmid was transferred into expression cells for expression, and the supernatant was collected and purified to obtain antibody protein samples, which were used in the following various examples.
  • the expression vector used is pCDNA3.1TM(+) (Invitrogen, the product number is V79020), and the expression cells are CHO-BAT-KF cells, and the level of fucosylation has been tested.
  • the sequences of the heavy chain and light chain of the positive control antibody 11D4 are from patent US8236930B2, the expression vector used is pCHO1.0 plasmid (purchased from Invitrogen), and the expression cells are CHO-S cells (purchased from Invitrogen).
  • the amino acid sequence of the antibody M-KF is shown in Table 1, the nucleic acid sequence of the antibody M-KF is shown in Table 2, and the sequence of the leader peptide (ie, the leader sequence) is in the frame in Table 2.
  • the anti-tumor efficacy of the antibody M-KF of the present invention was investigated in an OX40 humanized mouse model (purchased from Biocytograph). 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.
  • mice When the tumor grew to 119mm 3 , the mice were randomly divided into groups according to the tumor volume, 6 in each group.
  • the day of grouping is defined as D0 day, and the administration starts on D0 day; the grouping dosing schedule is shown in Table 3, and the administration dates are: day 0, day 3, day 6, day 9, day 12, day 15 sky.
  • Antibodies were administered intraperitoneally, according to the frequency of Q3D (once every 3 days), three dose groups of 1 mg/kg, 0.2 mg/kg and 0.04 mg/kg were administered 6 times in total.
  • the dosing schedule is shown in Table 3.
  • Tumors and body weight were measured twice weekly throughout the study, and mice were euthanized when tumors reached endpoints or when mice had 20% body weight loss.
  • 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 there is a significant difference.
  • TGItv inhibition rate relative to tumor volume
  • TGItv (1-(mean RTV administration group)/(mean RTV control group )) ⁇ 100%; mean RTV administration group: average RTV of administration group, mean RTV control group : average RTV of control group;
  • RTVn Vnt / Vn0 ;
  • Vnt the tumor volume of the mouse numbered n on the t day,
  • Vn0 the tumor volume of the mouse numbered n on the 0th day,
  • RTVn the numbered Relative tumor volume of n mice on day t.
  • N is the number of animals.
  • the middle and high doses of antibody M-KF can significantly inhibit tumor growth (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 the antibody M-KF.
  • Antibody M-KF was administered at doses of 0.5 mg/kg, 2.5 mg/kg and 10 mg/kg, 6 cynomolgus monkeys in each group, 3 males and 3 males, 18 in total; single intravenous infusion.
  • Cynomolgus monkeys were given a single intravenous infusion of antibody M-KF (0.5, 2.5 or 10 mg/kg), and all dose groups were well tolerated, with no obvious toxicity, and the maximum tolerated dose (MTD) was greater than 10 mg/kg.
  • M-KF maximum tolerated dose
  • the administration method was intravenous infusion of antibody M-KF (1, 5 and 30 mg/kg) or vehicle, once a week, for a total of 5 times, and the drug was stopped and recovered for 4 weeks; the administration time was Day 1, Day 8, Day 15, Day 22, Day 29; 10 cynomolgus monkeys in each group, 5 males and 5 males, a total of 40; 24 cynomolgus monkeys (each group 3 males and 3 females) were euthanized, and the remaining 16 cynomolgus monkeys (2 males and 2 females in each group) were euthanized after the recovery period.
  • the following indicators were evaluated: clinical observation, ophthalmological examination, body weight, food consumption, body temperature, hematology, blood coagulation, plasma biochemistry, immunoglobulin, lymphocyte immunophenotype, urine, macroscopic morphological observation, dirty instrument weight and histopathological examination.
  • Cynomolgus monkeys were infused with antibody M-KF (1, 5 or 30 mg/kg) by intravenous infusion once a week for 5 consecutive times, and the male and female animals at each dose could tolerate it well. cell percentage), transient elevation of IL-6, and minor histological changes in spleen/liver; therefore, the highest non-serious toxic dose (HNSTD) for this study was considered to be 30 mg/kg.
  • HNSTD non-serious toxic dose
  • Cmax ratio mean Cmax /corresponding mean Cmax in the 1 mg/kg dose group
  • AUC 0-t ratio mean AUC 0-t /corresponding AUC 0-t in the 1 mg/kg dose group
  • accumulation ratio AR Mean AUCo-t on Day 22 / Mean AUCo - t on Day 1.
  • Serum samples were collected, using a validated ECLA assay (electrochemiluminescence immunoassay for immunogenicity): after repeated intravenous infusions of antibody M-KF (1, 5, or 30 mg/kg), the individual positive rates were 90.0% (9 /10), 80.0% (8/10) and 20.0% (2/10), the overall positive rate of animals after administration was 63.3% (19/30).
  • ECLA assay electrochemical immunoassay for immunogenicity
  • This study is a multicenter, open-label, dose-escalation study to evaluate the safety, tolerability, pharmacokinetic (PK) characteristics, and preliminary clinical efficacy of antibody M-KF injection in patients with advanced malignant solid tumors.
  • Phase I clinical trials explore the maximum tolerated dose (MTD) or maximum administered dose (MAD), and provide recommended doses and reasonable dosing regimens for phase II or subsequent clinical studies.
  • MTD maximum tolerated dose
  • MAD maximum administered dose
  • This dose escalation study uses a "3+3"-based dose escalation rule to explore safe dose ranges.
  • Phase 1 0.01mg/kg group, 0.03mg/kg group, 0.1mg/kg group adopt accelerated titration method for dose escalation;
  • Phase 2 0.3mg/kg group, 1mg/kg group, 3mg/kg group according to standard Dose escalation studies were performed using the "3+3" rule.
  • 1 subject is included. If no DLT event is observed during the DLT evaluation period, it can directly enter the next group for dose escalation.
  • the accelerated escalation to (0.1mg /kg) dose group.
  • DLT evaluation period the first treatment cycle (from the first dose to 21 days after the dose).
  • DLT dose-limiting toxicity
  • PK pharmacokinetic
  • the parameters involved in PK are: for a single dose, parameters include C max , T max , T 1/2 , CL, Vd, Ke, MRT, AUC (0- ⁇ ) , AUC (0- ⁇ ) ; multiple doses When the drug is used, the parameters include C max,ss , C avg,ss , C min,ss , AUC (0- ⁇ ) ,ss, AUC( 0 - ⁇ ) ,ss, Tmax,ss, T 1/2 ,ss, CL, Vss, Ke, MRT, accumulation index (Rac), volatility index DF.
  • Objective Response Rate (ORR): the proportion of subjects with complete remission (CR) and partial remission (PR).
  • Duration of Response (DOR) DOR is defined as the time from the first assessment of the tumor as objective response (OR) to the first assessment as PD or PD (Progressive Disease) before death from any cause, reflecting ORR duration.
  • Progression-Free Survival (PFS): The time from the first administration to the occurrence of objective tumor progression or all-cause death (whichever occurs first). "MTD" is the highest dose level of DLT explored in a dose group in which ⁇ 1/6 of the subjects observed during the DLT evaluation period.
  • DLT Dose-Limiting Toxicity
  • AE adverse event
  • Grade 3 non-hematological toxicity including Grade 3 rash, nausea, vomiting or diarrhea lasting >3 days;

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Immunology (AREA)
  • Veterinary Medicine (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Hematology (AREA)
  • Oncology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

本发明公开了抗OX40抗体在治疗肿瘤或癌症中的应用，治疗方法包括向有需要的患者给药有效量的抗OX40抗体。

Description

抗OX40抗体在治疗肿瘤或癌症中的应用 技术领域
本发明属于生物医药领域，尤其涉及抗OX40抗体在治疗肿瘤或癌症中的应用。
背景技术
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。而免疫系统中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)。
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)。
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)。
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。
发明内容
本发明公开了抗OX40抗体或抗原结合片段用于治疗肿瘤或癌症的方法或用途。在一些实施方案中，抗OX40抗体或抗原结合片段剂用于治疗肿瘤或癌症。
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。
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。
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，或与SEQ ID NO:7所示序列具有至少80％同一性的序列，或与SEQ ID NO:7所示序列相比具有一或多个保守氨基酸取代的氨基酸序列；和/或
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，或与SEQ ID NO:8所示序列具有至少80％同一性的序列，或与SEQ ID NO:8所示序列相比具有一或多个保守氨基酸取代的氨基酸序列。
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。
在一些实施方案中，所述抗OX40抗体的重链包含SEQ ID NO:9所示的序列，或与SEQ ID NO:9所示序列具有至少80％同一性的序列，或与SEQ ID NO:9所示序列相比具有一或多个保守氨基酸取代的氨基酸序列；和/或
所述抗OX40抗体的轻链包含SEQ ID NO:10所示的序列，或与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或多个保守氨基酸取代的氨基酸序列。
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含有两条序列相同的重链和两条序列相同的轻链。
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)。
抗OX40抗体或抗原结合片段可以通过基因工程在CHO细胞或HEK293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。Protein A亲和柱和离子交换柱等方法可以用于纯化抗体蛋白。
在一些实施方案中，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，如PCT/CN2018/100008公开的CHO-BAT-KF细胞系。在一些实施方案中，抗体M-KF由CHO-BAT-KF细胞系表达。
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。在一些实施方案中，所述抗OX40抗体为抗体M-KF。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细胞系表达。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900mg。
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。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抗体或抗原结合片段，其中抗OX40抗体或抗原结合片段施用的有效量为约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-KF。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细胞系表达。
在一些实施方案中，可以将抗OX40抗体配制成药物组合物，并以适合于所选给药途径的多种形式向患者给药，给药途径例如肠胃外、静脉内(iv)、肌肉内、局部或皮下。在一些实施方案中，可以将抗OX40抗体静脉输注。抗OX40的给药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，以及所治疗的疾病和患者的状况(如年龄、性别、体重等)。
在一些实施方案中，每次施用的抗OX40抗体(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。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。在一些实施方案中，约1周、约2周、约3周或约4周给药1次。
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：每3周向有需要的患者给予约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的抗OX40抗体，或含此剂量抗OX40抗体的制剂。
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：每3周向有需要的患者给予约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的抗OX40抗体M，或含此剂量抗OX40抗体M的制剂。
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：每3周向有需要的患者给予约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的抗OX40抗体M-KF，或含此剂量抗OX40抗体M-KF的制剂。
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(或制剂)；其中，抗OX40抗体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)。剂量时间表和给药方式取决于某些患者群中的抗OX40抗体(或制剂)的获益风险评估和一般临床实践指南。
在一些实施方案中，患者每个治疗周期内抗OX40抗体施用的有效量为约0.6mg至900mg(或含此剂量抗OX40抗体的制剂)。
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(如抗体M或M-KF)的有效量为约0.6mg、约1mg、约1.8mg、约6mg、约10mg、约18mg、约30mg、约60mg、约100mg、约120mg、约180mg、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、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抗体的有效量为约0.6mg至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、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0.6mg至约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。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0.6mg、约1mg、约1.5mg、约1.8mg、约2mg、约2.9mg、约3.6mg、约4mg、约4.3mg、约5mg、约5.5mg、约6mg，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。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60mg至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60mg、约80mg、约90mg、约100mg、约120mg、约160mg、约180mg、约200mg、约243mg、约290mg、约355mg、约400mg、约425mg、约480mg、约500mg、约565mg、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，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600mg、约680mg、约 700mg、约750mg、约780mg、约800mg、约820mg、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。
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(如抗体M或M-KF)的有效量为约0.5mg至1mg，或含此剂量抗OX40抗体的制剂；比如约0.6mg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0.6mg，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。
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(如抗体M或M-KF)的有效量为约1.5mg至2.2mg，或含此剂量抗OX40抗体的制剂；比如约1.8mg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1.8mg，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(如抗体M或M-KF)的有效量为约4mg至8mg，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。
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(如抗体M或M-KF)的有效量为约50mg至76mg，或含此剂量抗OX40抗体的制剂；比如约60mg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60mg，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(如抗体M或M-KF)的有效量为约167mg至189mg，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180mg，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(如抗体M或M-KF)的有效量为约390mg至410mg，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。
在一些实施方案中，患者每个治疗周期内施用抗OX40抗体(如抗体M或M-KF)的有效量为约493mg至512mg，或含此剂量抗OX40抗体的制剂；比如约500mg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500mg，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(如抗体M或M-KF)的有效量为约588mg至606mg，或含此剂量抗OX40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OX40抗体的有效量为约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(如抗体M或M-KF)的有效量为约882mg至916mg，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。
在一些实施方案中，抗OX40抗体施用的有效量为约0.6mg至600mg每3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3周一次。在一些实施方案中，抗OX40抗体施用的有效量为约18mg、约60mg或约180mg每3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3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800mg或约900mg每3周一次。
在一些实施方案中，患者每个治疗周期内给药一次抗OX40抗体(或制剂)。在一些实施方案中，每个治疗周期内多次给药抗OX40抗体(或制剂)，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。
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个或4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。
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：每3周向有需要的患者给予约0.6mg至6mg、约6mg至60mg、约60mg至180mg、约180mg至600mg、约600mg至900mg，比如约0.6mg、约1.8mg、约6mg、 约18mg、约60mg、100mg、约180mg、约200mg、约300mg、约400mg、约500mg、约600mg或约900mg的抗OX40抗体，或含此剂量抗OX40抗体的制剂。在一些实施方案中，抗OX40抗体为抗体M。在一些实施方案中，抗OX40抗体为抗体M-KF。
在一些实施方案中，每3周一次给药抗OX40抗体约0.6mg。在一些实施方案中，每3周一次给药抗OX40抗体约1.8mg。在一些实施方案中，每3周一次给药抗OX40抗体约6mg。在一些实施方案中，每3周一次给药抗OX40抗体约18mg。在一些实施方案中，每3周一次给药抗OX40抗体约60mg。在一些实施方案中，每3周一次给药抗OX40抗体约180mg。在一些实施方案中，每3周一次给药抗OX40抗体约600mg。在一些实施方案中，每3周一次给药抗OX40抗体约900mg。
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，患者后的症状未得到预期缓解，再对患者给药约0.6mg至900mg抗OX40抗体，直至患者的症状得到缓解。
在一些实施方案中，抗OX40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)是输液方式进行给药。在一些实施方案中，抗OX40抗体(或制剂)是推注方式进行给药。
在一些实施方案中，抗OX40抗体(或制剂)是通过静脉内(i.v.)输液方式(即静脉输注)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。在一些实施方案中，静脉输液持续时间≥60分钟。
在一些实施方案中，抗OX40抗体(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、免疫治疗、激素治疗、靶向治疗、生物治疗和手术治疗等。在一些实施方案中，抗OX40抗体(或制剂)与其他肿瘤或癌症治疗剂联合治疗肿瘤或癌症，如激素、治疗肿瘤或癌症的抗体等。
另一方面，本发明公开了抗OX40抗体在制备用于治疗肿瘤或癌症的药物中的应用。在一些实施方案中，用于治疗肿瘤或癌症的药物包括抗OX40抗体。在一些实施方案中，抗OX40抗体为抗体M。在一些实施方案中，抗OX40抗体为抗体M-KF。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细胞系表达。
另一方面，本发明还公开了一种试剂盒，试剂盒包含抗OX40抗体(或制剂)和用于指导有需要患者给药抗OX40抗体(或制剂)的说明书。在一些实施方案中，抗 OX40抗体为抗体M。在一些实施方案中，抗OX40抗体为抗体M-KF。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细胞系表达。
另一方面，本发明还公开了包含抗OX40抗体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。适于注射用途的药物组合物包括无菌水性溶液(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，药物组合物还包括药学可接受的载体。在一些实施方案中，药学上可接受的载体可以包含抗细菌剂和/或抗真菌剂，如对羟基苯甲酸酯、氯代丁醇、苯酚、抗坏血酸、硫柳汞等来实现。在一些实施方案中，药学上可接受的载体可以包含等渗剂，如糖、多元醇(诸如甘露糖醇、山梨醇)、氯化钠。在一些实施方案中，药物组合物至少包含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的约2％至90％之间。这种治疗上有用的药物组合物中抗OX40抗体的量可以为给药的有效量。在一些实施方案中，抗OX40抗体为抗体M。在一些实施方案中，抗OX40抗体为抗体M-KF。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细胞系表达。
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抗体与药学上可接受的载体的混合方法是本领域通常已知的。
本发明抗OX40抗体(或制剂)可用于肿瘤或癌症的治疗。
附图说明
图1显示抗OX40抗体M-KF抑制肿瘤细胞的增殖；纵坐标表示肿瘤体积。
术语
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。
定义
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。
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组成的分子。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。
“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)。
“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。
“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。
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。
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。
“同一性”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。
“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。
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+Swi ssProtein+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。
“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)。
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。这四条链通过二 硫键以“Y”构型连接，其中轻链从“Y”口开始并延续通过可变区包围重链。
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(例如类Fab、类Fab'和类F(ab') 2)、类单链Fvs(scFv)。
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。
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(VL)和重链(VH)链部分的可变区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。
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。
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。
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。
“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。
“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。
“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。
“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。
如本文所用，术语“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，患者可能需要该方法和治疗。
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。
具体实施方式
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。
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。
实施例1抗体的制备方法
根据抗体的重链和轻链氨基酸序列构建轻链和重链的DNA序列。用PCR引物修饰DNA序列的5’和3’端，所述引物设计成为各链增加合适的前导序列，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。将上述重组质粒转入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。
其中，1)抗体M-KF制备过程中，采用的表达载体为pCDNA3.1TM(+)(Invitrogen公司，货号为V79020)，表达细胞为CHO-BAT-KF细胞，经测试岩藻糖基化水平为0；2)阳性对照抗体11D4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-KF的氨基酸序列见表1，抗体M-KF的核酸序列见表2，表2中框内为前导肽的序列(即前导序列)。
表1抗体M及抗体M-KF的氨基酸序列(按kabat进行划分)
Figure PCTCN2021131542-appb-000001
Figure PCTCN2021131542-appb-000002
表2抗体M及抗体M-KF的核酸序列
Figure PCTCN2021131542-appb-000003
Figure PCTCN2021131542-appb-000004
实施例2体外药效试验
1)肿瘤接种
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。
2)分组和给药
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；分组给药方案见表3，给药日期为：第0天、第3天、第6天、第9天、第12天、第15天。抗体腹腔给药，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3所示。
3)实验观察和数据采集
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)。
4)统计
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。
TGItv(相对肿瘤体积的抑制率)计算公式：
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；
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。
表3给药方案
Figure PCTCN2021131542-appb-000005
备注：N为动物只数。
如图1和表4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。
表4第24天的肿瘤体积的抑制率
Figure PCTCN2021131542-appb-000006
实施例3药代动力学和毒理试验
1)试验设计：抗体M-KF给药剂量为0.5mg/kg、2.5mg/kg和10mg/kg，每组6只食蟹猴，雌雄各3只，共18只；单次静脉输注。
a)药代动力学
于给药前(0h)，给药后2min、2h、6h、12h、24h、48h、72h、96h、120h、 168h、240h、336h、408h、504h和672h采集全血，分离血清；使用经过验证的ELISA方法分析各样本中药物的浓度，定量下限为30ng/mL。
各组动物血清中药物代谢动力学参数统计如表5所示，静脉输注抗体后，血清中抗体M-KF的浓度迅速达到峰值并以大致双相的方式下降；抗体M-KF的暴露量(平均C max和平均AUC 0-t)随剂量增加而增加，增加比近似剂量增加比例。
表5单次给药后，各组动物血清中药物代谢动力学参数
Figure PCTCN2021131542-appb-000007
b)毒性评价
所有动物在第30天进行解剖观察。试验期间，各组动物均未见死亡或濒死现象，各组动物的临床观察、体重、耗食量、血凝和血浆生化均未见明确与供试品相关的异 常改变。
食蟹猴单次静脉输注抗体M-KF(0.5、2.5或10mg/kg)，各剂量组均能很好耐受，未见明显毒性反应，最大耐受剂量(MTD)大于10mg/kg。
c)免疫原性
静脉输注抗体M-KF(0.5、2.5或10mg/kg)后，个体阳性率分别为100％(6/6)、100％(6/6)和83.3％(5/6)，食蟹猴的总体阳性率为94.4％(17/18)。
d)受体占位
于第-1天(0h)、第2天(给药后24h±1h)、第8天和第29天各采血1次。采用流式分析方法对样品进行分析。
各剂量组T淋巴细胞(CD3 +和CD3 +CD4 +)的OX40受体占位在第2天均接近饱和(RO率>80％)，第8天仍维持饱和状态；第29天时，2.5mg/kg和10mg/kg剂量组仍维持受体占位饱和状态，而0.5mg/kg剂量组降低至20％左右，与给药前(第-1天)基本持平。
2)试验设计：给药方式为静脉输注抗体M-KF(1、5和30mg/kg)或溶媒，每周给药1次，共5次，停药恢复4周；给药的时间为第1天、第8天、第15天、第22天、第29天；每组10只食蟹猴，雌雄各5只，共40只；给药结束后将24只食蟹猴(每组雌雄各3只)进行安乐死，剩余的16只食蟹猴(每组雌雄各2只)于恢复期结束后进行安乐死。试验期间，对下列指标进行评价：临床观察、眼科检查、体重、耗食量、体温、血液学、血凝、血浆生化、免疫球蛋白、淋巴细胞免疫表型、尿液、肉眼形态学观察、脏器重量和病理组织学检查。
a)毒性评价
食蟹猴每周1次连续5次静脉输注抗体M-KF(1、5或30mg/kg)，各剂量下雌雄动物均能较好耐受，各给药组主要可见NEUT(中性粒细胞百分率)下降、IL-6一过性升高以及脾脏/肝脏的轻微组织学改变；因此认为本试验的最高非严重毒性剂量(HNSTD)为30mg/kg。
b)毒代动力学
所有需分析的样品均使用已验证的ELISA方法进行分析。如表6所示，除个别动物外，各剂量组绝大多数动物在第1天和第22天给药后血清药物浓度迅速达峰(给药后2min)；第1天雌雄动物平均C max和平均AUC 0-t均随给药剂量增加而增加，增加比例与给药剂量增加比相当，雌雄动物无明显差别。
表6重复给药的毒代动力学参数
Figure PCTCN2021131542-appb-000008
注：C maxratio＝平均C max/1mg/kg剂量组中对应的平均C max；AUC 0-t ratio＝平均AUC 0-t/1mg/kg剂量组中对应的AUC 0-t；蓄积比AR＝第22天的平均AUC 0-t/第1天的平均AUC 0-t。
c)免疫原性
采集血清样品，采用已验证的ECLA分析方法(电化学发光免疫分析疫原性)：重复静脉输注抗体M-KF(1、5或30mg/kg)后，个体阳性率分别为90.0％(9/10)、80.0％(8/10)和20.0％(2/10)，给药后动物的总体阳性率为63.3％(19/30)。
d)给药局部刺激
食蟹猴连续4周静脉输注抗体M-KF(1、5或30mg/kg)后，各给药组动物给药部位均未见受试物相关的局部刺激反应。
实施例3体外溶血实验和组织交叉反应试验
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。
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。
实施例4抗体M-KF的临床研究
本研究是一项评价抗体M-KF注射液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。
本剂量递增研究采用基于“3+3”的剂量递增规则来探索安全剂量范围。第1阶段：0.01mg/kg组、0.03mg/kg组、0.1mg/kg组采用加速滴定方法进行剂量递增；第2阶段，0.3mg/kg组、1mg/kg组、3mg/kg组按标准“3+3”规则进行剂量递增研究。
三组采用加速滴定的剂量递增方案即队列规模＝1。首先纳入1例受试者，在DLT评估期内如果未观察到DLT事件，则可直接进入下一组组进行剂量递增，以此规则(“1+1”)类推，加速递增至(0.1mg/kg)剂量组。在加速递增过程中，若在某一组别观察到与研究药物相关的2级及以上毒性反应，则停止加速剂量递增，转为标准“3+3”剂量递增(即队列规模＝3)规则。
每3周以静脉输注给药1次，为1个周期(疗程)，输注时长可≥60分钟。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。持续治疗直到出现：疾病进展、或不可耐受的毒性而退出、或因无疗效获益而接受新的抗肿瘤治疗、或撤回知情同意等其它原因主动退出、或最长34个周期(约2年)，以先出现的为准。DLT评估期：为第1个治疗周期(从首次给药至给药后21天)。
耐受性评价指标涉及：剂量限制性毒性(DLT)事件及其发生率；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。
所有剂量组的受试者在治疗期间(前4个治疗周期)的规定时间点需要收集血样，每个时间点计划采2mL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：单次给药时，参数包括C max、T max、T 1/2、CL、Vd、Ke、MRT、AUC (0-τ)、AUC (0-∞)；多次给药时，参数包括C max,ss、C avg,ss、C min,ss、AUC (0-τ),ss、AU C(0- ∞),ss、Tmax,ss、T 1/2,ss、CL、Vss、Ke、MRT、蓄积指数(Rac)、波动指数DF。
临床有效性评价：客观缓解率(Objective Response Rate,ORR)：为完全缓解(CR)和部分缓解(PR)的受试者的比例。缓解持续时间(Duration of Response,DOR)：DOR定义为肿瘤第一次评估为客观缓解(Objective Response，OR)至第一次评估为PD或PD(Progressive Disease)前任何原因死亡的时间，反映ORR的持续时间。无进展生存期(Progression-Free Survival：PFS)：为从首次给药至出现肿瘤客观进展或全因死亡的时间(以先发生者为准)。“MTD”为某一剂量组在DLT评估期内观察到≤1/6的受试者中探索到的DLT的最高剂量水平。
“剂量限制性毒性(DLT)”为在DLT观察期发生的不良事件(AE)并被认为至少可能与研究药物相关，具体如下：
■血液学毒性：
·4级血液学毒性；
·4级血小板减少症；有出血倾向的或需要输注血小板的3级血小板减少症；
·≥3级的中性粒细胞减少伴发热；
■非血液学毒性：
·4级非血液学毒性；
·非临床实验室检查发现的：3级非血液学毒性，包括3级的持续时间＞3天的皮疹、恶心、呕吐或腹泻；
·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)异常持续超过1星期导致受试者在第1周期停止治疗的任何药物相关AE，4)导致第2周期治疗延迟≥2周的任何药物相关AE。

Claims (11)

  1. 一种用于治疗肿瘤或癌症的方法，其特征在于，所述方法包括：向有需要的患者给药有效量的抗OX40抗体或抗原结合片段；
    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。
  2. 如权利要求1所述的方法，其特征在于，所述抗OX40抗体或抗原结合片段包含SEQ ID NO:7所示的重链可变区，或与SEQ ID NO:7所示序列具有至少80％同一性的序列，或与SEQ ID NO:7所示序列相比具有一或多个保守氨基酸取代的氨基酸序列；和/或
    所述抗OX40抗体或抗原结合片段包含SEQ ID NO:8所示的轻链可变区，或与SEQ ID NO:8所示序列具有至少80％同一性的序列，或与SEQ ID NO:8所示序列相比具有一或多个保守氨基酸取代的氨基酸序列。
  3. 如权利要求1或2所述的方法，其特征在于，所述抗OX40抗体的重链包含SEQ ID NO:9所示的序列，或与SEQ ID NO:9所示序列具有至少80％同一性的序列，或与SEQ ID NO:9所示序列相比具有一或多个保守氨基酸取代的氨基酸序列；和/或
    所述抗OX40抗体的轻链包含SEQ ID NO:10所示的序列，或与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或多个保守氨基酸取代的氨基酸序列。
  4. 如权利要求1-3任一项所述的方法，其特征在于，所述抗OX40抗体的岩藻糖基化水平为0-10％。
  5. 如权利要求1-3任一项所述的方法，其特征在于，所述抗OX40抗体由α-(1,6)-岩藻糖基转移酶基因敲除的细胞表达。
  6. 如权利要求1-5任一项所述的方法，其特征在于，所述肿瘤或癌症包括急性淋巴细胞性白血病、急性骨髓性白血病、慢性淋巴细胞性白血病、慢性骨髓性白血病、骨髓性增生疾病/肿瘤、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、滤泡性淋巴瘤、多发性骨髓瘤、巨细胞骨髓瘤、重链骨髓瘤、轻链或本斯-琼斯骨髓瘤、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。
  7. 如权利要求1-6任一项所述的方法，其特征在于，每个治疗周期内抗OX40抗体给药的有效量为0.6mg至900mg。
  8. 如权利要求7所述的方法，其特征在于，一个治疗周期为1周、2周、3周、或4周。
  9. 如权利要求1-8任一项所述的方法，其特征在于，所述抗OX40抗体的每次给药量为0.01mg/kg-25mg/kg。
  10. 如权利要求1-9任一项所述的方法，其特征在于，给药方式为静脉注射或皮下注射。
  11. 一种试剂盒，其特征在于，包含抗OX40抗体或抗原结合片段和用于指导有需要患者给药抗OX40抗体或抗原结合片段的说明书；
    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。
PCT/CN2021/131542 2020-11-19 2021-11-18 抗ox40抗体在治疗肿瘤或癌症中的应用 Ceased WO2022105839A1 (zh)

Priority Applications (3)

Application Number Priority Date Filing Date Title
CN202180073819.9A CN116507365A (zh) 2020-11-19 2021-11-18 抗ox40抗体在治疗肿瘤或癌症中的应用
US18/037,401 US20230416386A1 (en) 2020-11-19 2021-11-18 Use of anti-ox40 antibody in treatment of tumor or cancer
EP21893986.6A EP4248995A4 (en) 2020-11-19 2021-11-18 USE OF ANTI-OX40 ANTIBODIES FOR THE TREATMENT OF TUMORS OR CANCERS

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202011301761.2A CN114515335A (zh) 2020-11-19 2020-11-19 抗ox40抗体在治疗肿瘤或癌症中的应用
CN202011301761.2 2020-11-19

Publications (1)

Publication Number Publication Date
WO2022105839A1 true WO2022105839A1 (zh) 2022-05-27

Family

ID=81594769

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/131542 Ceased WO2022105839A1 (zh) 2020-11-19 2021-11-18 抗ox40抗体在治疗肿瘤或癌症中的应用

Country Status (4)

Country Link
US (1) US20230416386A1 (zh)
EP (1) EP4248995A4 (zh)
CN (2) CN114515335A (zh)
WO (1) WO2022105839A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024193516A1 (zh) * 2023-03-17 2024-09-26 百奥泰生物制药股份有限公司 抗ox40抗体在治疗炎症或免疫性疾病中的应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113429483A (zh) * 2020-03-23 2021-09-24 百奥泰生物制药股份有限公司 一种免疫细胞激活剂的开发及应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8236930B2 (en) 2007-12-14 2012-08-07 Pfizer Inc. Binding molecules to the human OX40 receptor
US20170182156A1 (en) * 2014-04-03 2017-06-29 Augusta University Research Institute, Inc. Methods for enhancing the efficacy of a tumor-directed immune response
WO2017130076A1 (en) * 2016-01-25 2017-08-03 Pfizer Inc. Combination of an ox40 agonist and a 4-1bb agonist monoclonal antibody for treating cancer
WO2020063660A1 (zh) * 2018-09-26 2020-04-02 江苏恒瑞医药股份有限公司 抗ox40抗体、其抗原结合片段及其医药用途

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI461436B (zh) * 2005-11-25 2014-11-21 Kyowa Hakko Kirin Co Ltd 人類ｃｄ１３４（ｏｘ４０）之人類單株抗體及其製造及使用方法
SG11201607969XA (en) * 2014-03-31 2016-10-28 Genentech Inc Anti-ox40 antibodies and methods of use
BR112019012328A2 (pt) * 2016-12-15 2019-11-19 Abbvie Biotherapeutics Inc anticorpos anti-ox40 e seus usos
CN113429483A (zh) * 2020-03-23 2021-09-24 百奥泰生物制药股份有限公司 一种免疫细胞激活剂的开发及应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8236930B2 (en) 2007-12-14 2012-08-07 Pfizer Inc. Binding molecules to the human OX40 receptor
US20170182156A1 (en) * 2014-04-03 2017-06-29 Augusta University Research Institute, Inc. Methods for enhancing the efficacy of a tumor-directed immune response
WO2017130076A1 (en) * 2016-01-25 2017-08-03 Pfizer Inc. Combination of an ox40 agonist and a 4-1bb agonist monoclonal antibody for treating cancer
WO2020063660A1 (zh) * 2018-09-26 2020-04-02 江苏恒瑞医药股份有限公司 抗ox40抗体、其抗原结合片段及其医药用途

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
AUSUBEL ET AL.: "Current Protocols in Molecular Biology", 2007
BUCHAN SARAH L.; ROGEL ANNE; AL-SHAMKHANI AYMEN: "The immunobiology of CD27 and OX40 and their potential as targets for cancer immunotherapy", BLOOD, AMERICAN SOCIETY OF HEMATOLOGY, US, vol. 131, no. 1, 4 January 2018 (2018-01-04), US , pages 39 - 48, XP086691630, ISSN: 0006-4971, DOI: 10.1182/blood-2017-07-741025 *
CHOTHIA ET AL., J. MOL. BIOL., vol. 196, 1987, pages 901 - 917
GRAMAGLIA I ET AL., J. IMMUNOL., vol. 165, 2000, pages 3043 - 3050
J. WILLOUGHBY ET AL., MOLECULAR IMMUNOLOGY, vol. 83, 2017, pages 13 - 22
KABAT ET AL.: "Sequence of Proteins of Immunological Interest", U.S. DEPT. OF HEALTH AND HUMAN SERVICES, 1983
KABAT ET AL.: "Sequences of Proteins of Immunological Interest", U.S. DEPT. OF HEALTH AND HUMAN SERVICES, 1983
KUANG ZHIHUI; JING HUA; WU ZHIHAI; WANG JIE; LI YIMING; NI HAIQING; ZHANG PAN; WU WEIWEI; WU MIN; ZHOU SHUAIXIANG; QIU XUAN; WU DO: "Development and characterization of a novel anti-OX40 antibody for potent immune activation", CANCER IMMUNOLOGY IMMUNOTHERAPY, SPRINGER, BERLIN/HEIDELBERG, vol. 69, no. 6, 20 February 2020 (2020-02-20), Berlin/Heidelberg , pages 939 - 950, XP037127842, ISSN: 0340-7004, DOI: 10.1007/s00262-020-02501-2 *
LINDSAY K ET AL., IMMUNITY, vol. 44, no. 5, 2016, pages 1005 - 1019
PATERSON DJ ET AL., MOL. IMMUNOL.,, vol. 24, 1987, pages 1281 - 1290

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024193516A1 (zh) * 2023-03-17 2024-09-26 百奥泰生物制药股份有限公司 抗ox40抗体在治疗炎症或免疫性疾病中的应用

Also Published As

Publication number Publication date
CN114515335A (zh) 2022-05-20
EP4248995A1 (en) 2023-09-27
EP4248995A4 (en) 2024-12-11
US20230416386A1 (en) 2023-12-28
CN116507365A (zh) 2023-07-28

Similar Documents

Publication Publication Date Title
AU2012314390B2 (en) Anti-ICAM-1 antibodies to treat multiple-myeloma related disorders
JP7736783B2 (ja) エンゲージャを含む薬物治療剤の開発及び応用
WO2022184067A1 (zh) 抗tigit抗体在联合用药中的应用
WO2018232188A1 (en) Anti-l1-cam antibodies and uses thereof
WO2022242758A1 (zh) 抗cd73抗体及其应用
EP3746123A1 (en) Antibodies to galectin-3 and methods of use thereof
WO2021129775A1 (zh) 抗ctla-4单克隆抗体及其制备方法与应用
EP4382540A1 (en) Use of anti-pd-l1/cd47 bispecific antibody in treatment of diseases
TWI883241B (zh) 抗pd-l1抗體及其應用
WO2022105839A1 (zh) 抗ox40抗体在治疗肿瘤或癌症中的应用
CN116262790A (zh) 嵌合的抗cd27抗体组合物及其应用
IL293118A (en) Methods of cancer treatment using anti-ox40 antibodies in combination with anti-tigit antibodies
WO2023001118A1 (zh) 抗ox40抗体在联合用药中的应用
WO2022242757A1 (zh) 抗pd-1抗体的应用
CN114641500B (zh) 使用抗ox40抗体与抗tim3抗体的组合治疗癌症的方法
WO2022022526A1 (zh) 抗her2抗体及其应用
WO2021098749A1 (en) Methods of cancer treatment with anti-ox40 antibody in combination with radiation
WO2022184068A9 (zh) 抗tigit抗体在治疗肿瘤或癌症中的应用
CN121668303A (zh) 抗pd-1抗体在联合用药中的应用
CN117618555A (zh) 抗tigit抗体和抗ctla-4抗体联合用于治疗肿瘤中的应用
WO2023105528A1 (en) Antibodies specific to ceacam1
HK40067102A (zh) 抗her2抗体及其应用
WO2024193516A1 (zh) 抗ox40抗体在治疗炎症或免疫性疾病中的应用
WO2025195291A1 (zh) 治疗肿瘤的方法和用途
WO2024044675A1 (en) Methods of cancer treatment using anti-pd1 antibodies in combination with anti-tim3 antibodies

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21893986

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 202180073819.9

Country of ref document: CN

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2021893986

Country of ref document: EP

Effective date: 20230619