WO2024046389A1 - Combined use of anti-tigit antibody and anti-ctla-4 antibody in treating tumor - Google Patents

Combined use of anti-tigit antibody and anti-ctla-4 antibody in treating tumor Download PDF

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WO2024046389A1
WO2024046389A1 PCT/CN2023/115904 CN2023115904W WO2024046389A1 WO 2024046389 A1 WO2024046389 A1 WO 2024046389A1 CN 2023115904 W CN2023115904 W CN 2023115904W WO 2024046389 A1 WO2024046389 A1 WO 2024046389A1
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seq
amino acid
acid sequence
antibody
antigen
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PCT/CN2023/115904
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French (fr)
Chinese (zh)
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黄俊杰
吴精博
张少榆
黄贤明
李胜峰
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百奥泰生物制药股份有限公司
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Publication of WO2024046389A1 publication Critical patent/WO2024046389A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • 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
    • 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
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention belongs to the field of biomedicine, and specifically relates to the use of anti-TIGIT antibodies and anti-CTLA-4 antibodies in combination to treat tumors.
  • TIGIT T cell immunoreceptor with Ig and ITIM domains
  • Ig immunoglobulin
  • ITIM tyrosine inhibitor motif
  • the TIGIT structure is shown to contain an extracellular immunoglobulin domain, a type I transmembrane region and two ITIM motifs.
  • TIGIT is part of a costimulatory network, which mainly consists of the activating receptor CD226 and the inhibitory receptor TIGIT on T cells, as well as the ligand CD155 (also known as CD155) expressed on the surface of APCs, tumor cells, and infected cells.
  • TIGIT a poliovirus receptor protein encoded by the PVR gene in humans
  • CD112 CD112.
  • the binding of TIGIT to PVR or CD112 will cause the phosphorylation of Tyr225 in the cytoplasm of TIGIT, and TIGIT will bind to the cell adaptive growth factor receptor binding protein 2 (GRB2).
  • GRB2 can recruit SHIP1 to inhibit phosphatidylinositol trikinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling.
  • PI3K phosphatidylinositol trikinase
  • MAPK mitogen-activated protein kinase
  • phosphorylated TIGIT recruits SHIP1 through beta-arrestin2 ( ⁇ -arrestin2) and blocks the autoubiquitination of TNF receptor-associated factor 6 (TRAF6) and disrupts nuclear factor KB (NF-KB) activation.
  • ⁇ -arrestin2 beta-arrestin2
  • NF-KB nuclear factor KB
  • Ser329 and Tyr322 of the intracellular domain of CD226 are phosphorylated; Ser329 phosphorylation promotes the activation of protein kinase (PKC) and the mutual binding of CD226 to lymphocyte-associated antigen 1 (LFA1). LFA1 is then used for TYN-mediated Tyr322 phosphorylation and CD226-mediated downstream signaling. A series of signal transduction ultimately leads to the activation of T cell or NK cell function and promotes the production of cytokines.
  • PDC protein kinase
  • LFA1 lymphocyte-associated antigen 1
  • TIGIT molecules present on the surface of T cells or NK cells There is also an interaction between TIGIT molecules present on the surface of T cells or NK cells and CD226 molecules, which means that TIGIT molecules can directly disrupt CD226 to form normal dimers, thereby destroying the normal physiological functions of CD226.
  • TIGIT and CD226 are like two ends of a scale. Through the fulcrum of PVR, they subtly regulate the body's immune function through the transmission of co-stimulatory and co-inhibitory signals.
  • Cytotoxic T lymphocyte antigen-4 (CTLA-4) is a key inhibitory receptor that affects T cell function and plays a key role in the initiation phase of immune response (Scalapino KJ1, Daikh DI. (2008), Immunol Rev. 223:143-155).
  • TCR T cell receptor
  • MHC class I or class II antigen-presenting cells the signal on the TCR is amplified and counteracted by costimulatory molecules.
  • CD28 on T cells binds to B7-1 (CD80) and B7-2 (CD86) on antigen-presenting cells, it sends a signal that requires full activation of T cells.
  • This CD28/B7 combination leads to an increase in the production of interleukin 2 and other stimulatory cytokines, increases metabolism, promotes cell cycle progression, upregulates cell survival genes, and ultimately enables T cells to proliferate and differentiate.
  • CD28 binds and causes T cell proliferation
  • CTLA-4 is transported and expressed on the surface of T cells (Linsley PS, et al. (1996), Immunity. 4(6):535-543). The stronger the activation signal through the TCR, the more CTLA-4 is transported and expressed. While on the cell surface, CTLA-4's inhibitory signal is propagated (Egen JG, Allison JP. (2002), Immunity. 16(1):23-25).
  • CTLA-4 has a higher affinity for B7 than CD28 and blocks further coactivation (Krummel MF, Allison JP. (1995), J Exp Med. 182(2):458-465). Moreover, cells expressing CTLA-4 can capture and degrade B7-1 and B7-2 through endocytosis (Qureshi OS, et al. (2011), Science. 332 (6029): 600-603).
  • the object of the present invention is to provide a combined drug and its application in treating tumors.
  • the combination includes an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the combination includes a therapeutically effective amount of an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
  • the present invention provides a pharmaceutical combination for treating tumors, including an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
  • the invention provides anti-TIGIT antibodies or antigen-binding fragments and anti-CTLA-4 antibodies or antigen-binding fragments in combination for the treatment of tumors.
  • the invention provides a method of treating tumors comprising administering to a patient in need thereof an effective amount of an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
  • the invention provides the use of an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or an antigen-binding fragment in the preparation of a medicament for treating tumors in combination with an anti-CTLA-4 antibody or antigen-binding fragment.
  • an anti-TIGIT antibody eg, antibody h10D8OF or h10D8OFKF
  • an antigen-binding fragment in the preparation of a medicament for treating tumors in combination with an anti-CTLA-4 antibody or antigen-binding fragment.
  • the invention provides the use of an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment in the preparation of a medicament for treating tumors.
  • an anti-TIGIT antibody eg, antibody h10D8OF or h10D8OFKF
  • an anti-CTLA-4 antibody or antigen-binding fragment in the preparation of a medicament for treating tumors.
  • the invention provides use of an anti-CTLA-4 antibody or antigen-binding fragment in the preparation of a medicament for treating tumors in combination with an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment.
  • an anti-TIGIT antibody eg, antibody h10D8OF or h10D8OFKF
  • the invention provides the use of an anti-CTLA-4 antibody or antigen-binding fragment and an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment in the preparation of a medicament for treating tumors.
  • an anti-CTLA-4 antibody or antigen-binding fragment and an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment in the preparation of a medicament for treating tumors.
  • an anti-TIGIT antibody eg, antibody h10D8OF or h10D8OFKF
  • the invention provides anti-TIGIT antibodies (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragments and anti-CTLA-4 antibodies or antigen-binding fragments in the preparation of pharmaceutical compositions for treating tumors.
  • anti-TIGIT antibodies eg, antibody h10D8OF or h10D8OFKF
  • antigen-binding fragments and anti-CTLA-4 antibodies or antigen-binding fragments in the preparation of pharmaceutical compositions for treating tumors.
  • the invention provides a kit comprising an anti-TIGIT antibody and a package insert containing instructions for using the anti-TIGIT antibody in combination with an anti-CTLA-4 antibody to treat tumors.
  • the invention provides a kit comprising an anti-CTLA-4 antibody and a package insert containing instructions for using the anti-CTLA-4 antibody in combination with an anti-TIGIT antibody to treat tumors.
  • the present invention provides a kit comprising a medicament of an anti-TIGIT antibody and an anti-CTLA-4 antibody, and a package insert containing instructions for using the medicament of an anti-TIGIT antibody and an anti-CTLA-4 antibody. Instructions for treating tumors.
  • the present invention provides a kit comprising a medicament of an anti-TIGIT antibody and a package insert containing instructions for using the medicament of an anti-TIGIT antibody in combination with an anti-CTLA-4 antibody to treat tumors.
  • the present invention provides a kit comprising a medicament of an anti-CTLA-4 antibody and a package insert containing instructions for using the medicament of an anti-CTLA-4 antibody in combination with an anti-TIGIT antibody to treat tumors. .
  • the present invention provides a kit comprising an anti-TIGIT antibody or antigen-binding fragment (or preparation), an anti-CTLA-4 antibody or antigen-binding fragment (or preparation) and a method for instructing patients in need to administer anti-TIGIT Instructions for antibodies or antigen-binding fragments (or formulations) and anti-CTLA-4 antibodies or antigen-binding fragments (or formulations).
  • the anti-TIGIT 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 SEQ ID NO: 4, LCDR2 shown in SEQ ID NO: 5, and LCDR3 shown in SEQ ID NO: 6.
  • the anti-TIGIT 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 anti-TIGIT antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:7, or is at least 80% identical to the sequence set forth in SEQ ID NO:7. A unique amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:7.
  • the light chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:8, or is at least 80% identical to the sequence set forth in SEQ ID NO:8.
  • the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:7, or is at least 80% identical to the sequence set forth in SEQ ID NO:7.
  • the light chain variable region of the anti-TIGIT antibody or antigen-binding fragment includes SEQ ID NO:8
  • the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:7
  • the light chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises SEQ ID NO:7 The amino acid sequence shown in ID NO:8.
  • the heavy chain of the anti-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 9, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 9, or is identical to the sequence set forth in SEQ ID NO: 9.
  • the sequence shown in SEQ ID NO:9 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
  • the light chain of the anti-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 10, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 10, or is identical to the sequence set forth in SEQ ID NO: 10.
  • the sequence shown in SEQ ID NO:10 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
  • the heavy chain of the anti-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 9, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 9, or is identical to the sequence set forth in SEQ ID NO: 9.
  • the sequence shown in SEQ ID NO:9 is an amino acid sequence with one or more conservative amino acid substitutions;
  • the light chain of the anti-TIGIT antibody includes the amino acid sequence shown in SEQ ID NO:10, or is identical to the amino acid sequence shown in SEQ ID NO:10
  • the anti-TIGIT antibody is an antibody h10D8OF or h10D8OFKF, the heavy chain of which comprises the amino acid sequence shown in SEQ ID NO:9, and the light chain of which comprises the amino acid sequence of SEQ ID NO:10.
  • antibody h10D8OF or h10D8OFKF contains two sequence-identical heavy chains and two sequence-identical light chains.
  • the anti-TIGIT antibody eg, antibody h10D8OFKF
  • antigen-binding fragment has a fucosylation level of 0-10%. In some embodiments, the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment has a fucosylation level of 0-5%.
  • the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment has a fucosylation level 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 The range between (inclusive) or any value therein.
  • the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment does not bind fucose.
  • the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
  • the anti-CTLA-4 antibody or antigen-binding fragment at least comprises HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, HCDR3 shown in SEQ ID NO: 13, SEQ One or more of LCDR1 shown in ID NO:14, LCDR2 shown in SEQ ID NO:15, and LCDR3 shown in SEQ ID NO:16.
  • the anti-CTLA-4 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, HCDR3 shown in SEQ ID NO: 13, SEQ ID LCDR1 shown in NO:14, LCDR2 shown in SEQ ID NO:15 and LCDR3 shown in SEQ ID NO:16.
  • the anti-CTLA-4 antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO: 17, or has an amino acid sequence of at least 80% compared to the sequence set forth in SEQ ID NO: 17. % identical amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:17.
  • the light chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO: 18, or has an amino acid sequence of at least 80% compared to the sequence set forth in SEQ ID NO: 18. % identity of the amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:18.
  • the heavy chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO: 17, or has an amino acid sequence of at least 80% compared to the sequence set forth in SEQ ID NO: 17. % identical amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 17; the light chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment includes SEQ The amino acid sequence shown in ID NO:18, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:18, or one or more conserved amino acid sequences compared with the sequence shown in SEQ ID NO:18 Amino acid sequence of amino acid substitutions.
  • the heavy chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence shown in SEQ ID NO: 17, and the light chain of the anti-CTLA-4 antibody or antigen-binding fragment can The variable region contains the amino acid sequence shown in SEQ ID NO:18.
  • the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 19, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 19, Or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:19.
  • the light chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO:20, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO:20, Or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:20.
  • the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 19, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 19, or with SEQ
  • the sequence shown in ID NO: 19 is compared to the amino acid sequence having one or more conservative amino acid substitutions
  • the light chain of the anti-CTLA-4 antibody includes the amino acid sequence shown in SEQ ID NO: 20, or is the same as SEQ ID NO: 20
  • the sequence shown is an amino acid sequence that has at least 80% identity compared to the sequence shown in SEQ ID NO: 20, or an amino acid sequence that has one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 20.
  • the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 19
  • the light chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 20 sequence.
  • an anti-CTLA-4 antibody contains two sequence-identical heavy chains and two sequence-identical light chains.
  • the anti-CTLA-4 antibody or antigen-binding fragment has a total amount of high-mannose glycoforms ⁇ 5% and/or a total amount of sialylated glycoforms ⁇ 3%.
  • 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 of the endpoints) or any value therein.
  • the anti-CTLA-4 antibody or antigen-binding fragment has a total amount of sialylated glycoforms of about 0.1%, 0.2%, about 0.36%, about 0.8%, about 1.5%, about 2.2%, about 2.7 %, about 2.9%, 2.99%, or a range between any two of these values (inclusive of the endpoints) or any value therein.
  • the anti-CTLA-4 antibody or antigen-binding fragment has a total amount of high mannose glycoforms ⁇ 2% and/or a total amount of sialylated glycoforms ⁇ 1%.
  • 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 anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level 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 between any two of these values Range (including endpoints) or any value therein.
  • the anti-CTLA-4 antibody or antigen-binding fragment does not bind fucose. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
  • the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-10%, a total high-mannose glycoform ⁇ 5% and/or a total sialylated glycoform ⁇ 5%. 3%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%, a total high-mannose glycoform ⁇ 5% and/or a total sialylated glycoform ⁇ 3%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of about 0%, a total high-mannose glycoform ⁇ 5% and/or a total sialylated glycoform ⁇ 3 %.
  • the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-10%, a total high-mannose glycoform ⁇ 3% and/or a total sialylated glycoform ⁇ 3%. 2%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%, a total high-mannose glycoform ⁇ 3% and/or a total sialylated glycoform ⁇ 3%. 2%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of about 0%, a total high-mannose glycoform ⁇ 3% and/or a total sialylated glycoform ⁇ 2 %.
  • the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-10%, high glycerol The total amount of unglucose glycoforms is ⁇ 2% and/or the total amount of sialylated glycoforms is ⁇ 1%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%, a total high-mannose glycoform ⁇ 2% and/or a total sialylated glycoform ⁇ 2%. 1%.
  • the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of about 0%, a total amount of high-mannose glycoforms ⁇ 2%, and/or a total amount of sialylated glycoforms ⁇ 1 %.
  • the anti-CTLA-4 antibody or antigen-binding fragment is ipilimumab (Yervoy TM or a biosimilar thereof).
  • Antibodies can be expressed in host cells through genetic engineering and obtained through purification; purification can be performed using conventional methods, such as centrifuging the cell suspension and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as ProteinA affinity columns and ion exchange columns can be used to purify antibodies.
  • a hypofucosylated or afucosylated anti-TIGIT antibody eg, antibody h10D8OFKF
  • antigen-binding fragment or an anti-CTLA-4 antibody or antigen-binding fragment is composed of ⁇ -(1,6) - Expression from a fucosyltransferase gene knockout cell line, such as expression from ⁇ -(1,6)-fucosyltransferase gene knockout CHO-K1 cells.
  • the invention also provides pharmaceutical compositions comprising an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
  • the pharmaceutical composition is a pharmaceutical composition suitable for injection, such as a push-type pharmaceutical composition or an infusion (drip)-type pharmaceutical composition.
  • the pharmaceutical composition contains at least 0.1% anti-TIGIT antibody or antigen-binding fragment and 0.1% anti-CTLA-4 antibody or antigen-binding fragment.
  • the percentage of antibody can vary and can be between about 2% and about 90% by weight of a given dosage form.
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment in this therapeutically useful pharmaceutical composition can be administered in effective amounts.
  • the present invention also provides a method for preparing the above-mentioned pharmaceutical composition: respectively, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment described herein are combined with a pharmaceutically acceptable suitable for injection.
  • a pharmaceutically acceptable suitable for injection such as water for injection, physiological saline, etc.
  • the above pharmaceutical composition is prepared by: combining the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment described herein with a pharmaceutically acceptable carrier suitable for injection (e.g., injection Mix with water, saline, etc.).
  • a pharmaceutically acceptable carrier suitable for injection e.g., injection Mix with water, saline, etc.
  • the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-50:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 5-25:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-15:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-10:1.
  • the mass ratio of the anti-TIGIT antibody or antigen-binding fragment to the anti-CTLA-4 antibody or antigen-binding fragment is about 5:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 15-30:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 20-30:1. In some implementations In this case, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is approximately 25:1.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 1200 mg, about 0.5 mg to 1000 mg, about 5 mg to 500 mg, about 5 mg to 100 mg, about 10 mg to 100 mg, about 10 mg to 50 mg. , or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7mg, about 8mg, about 9mg, about 10mg, about 12mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300mg, About 330 mg, about 380 mg, about 400 mg, about 480 mg, about 500 mg, about 580 mg, about 600 mg, about 800 mg, about 900 mg, about 1000 mg, about 1200 mg, or a range between any two of these values (inclusive of the endpoints), or Any of these values, or a formulation containing this dose of anti-TIGIT antibody or antigen-binding fragment.
  • the 480 mg about 500 mg,
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg to 20 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg/kg, about 0.05 mg/kg to 1 mg/kg, about 0.1 mg/kg to 1 mg/kg, or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg, about 0.8mg/kg, about 0.9mg/kg, about 1mg/kg, about 2mg/kg, about 3mg/kg, about 4mg/kg, about 5mg/kg, about 6mg/kg, about 7mg/kg, about 8mg/kg , about 9 mg/kg, about 10 mg/kg, about 12 mg/kg, about 14 mg/kg, about 15 mg/kg, about 18 mg/kg, about 20 mg/kg, or a range between any two of these values (including endpoint) or any value therein, or a formulation containing such a dose of anti-TIGIT antibody or antigen-binding fragment.
  • the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 1200 mg, about 0.5 mg to 1000 mg, about 5 mg to 500 mg, about 5 mg to 100 mg, about 10 mg to 100 mg, about 10 mg per treatment cycle. to 50 mg, or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg per treatment cycle.
  • the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF.
  • the anti-TIGIT antibody or antigen-binding fragment is administered per treatment cycle at a dose of about 0.01 mg/kg to 20 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg/kg , about 0.05 mg/kg to 1 mg/kg, about 0.1 mg/kg to 1 mg/kg, or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg per treatment cycle.
  • the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF.
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 600 mg, about 0.5 mg to 600 mg, about 1 mg to 100 mg, about 1 mg to 50 mg, about 1 mg to 10 mg, or containing Preparation of anti-CTLA-4 antibodies or antigen-binding fragments at this dose.
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.5 mg, about 0.8 mg, about 1 mg, about 2 mg per administration.
  • the anti-CTLA-4 antibody is Antibody 1.
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg to 10 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg/kg per administration , about 0.02 mg/kg to 1 mg/kg, about 0.02 mg/kg to 0.5 mg/kg, about 0.02 mg/kg to 0.2 mg/kg, or preparations containing anti-CTLA-4 antibodies or antigen-binding fragments at this dose.
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg/kg, about 0.04 mg/kg, about 0.05 mg/kg per administration.
  • the anti-CTLA-4 antibody is Antibody 1.
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 600 mg, about 0.5 mg to 600 mg, about 1 mg to 100 mg, about 1 mg to 50 mg, about 1 mg to 10 mg per treatment cycle, or preparations containing anti-CTLA-4 antibodies or antigen-binding fragments at this dose.
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg per treatment cycle mg, about 0.2mg, about 0.3mg, about 0.5mg, about 0.8mg, about 1mg, about 2mg, about 3mg, about 4mg, about 5mg, about 6mg, about 7mg, about 8mg, about 9mg, about 10mg, about 12mg , about 20mg, about 30mg, about 50mg, about 60mg, about 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300mg, about 330mg, about 380mg, about 400mg, about 434mg, about 480mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, or a range between any two of these values (inclusive of the endpoints) or any value therein, or a formulation containing such
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg to 10 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg per treatment cycle. /kg, about 0.02 mg/kg to 1 mg/kg, about 0.02 mg/kg to 0.5 mg/kg, about 0.02 mg/kg to 0.2 mg/kg, or preparations containing anti-CTLA-4 antibodies or antigen-binding fragments at this dose .
  • the anti-CTLA-4 antibody or antigen-binding fragment is administered per treatment cycle at a dose of about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg/kg, about 0.04 mg/kg, about 0.05 mg /kg, about 0.06mg/kg, about 0.07mg/kg, about 0.08mg/kg, about 0.1mg/kg, about 0.3mg/kg, about 0.5mg/kg, about 0.8mg/kg, about 0.9mg/kg , about 1mg/kg, about 2mg/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 , or a range between any two of these values (inclusive of the endpoints) or any value therein, or a formulation containing such a dose of anti-CTLA-4 antibody or antigen-binding fragment.
  • the invention provides a method of treating tumors, the method comprising: administering to a patient in need thereof about 0.05 mg to 1200 mg, about 0.5 mg to 1000 mg, about 5 mg to 500 mg, about 5 mg to 100 mg, About 10 mg to 100 mg, about 10 mg to 50 mg, such as about 0.05 mg, about 0.08 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg , about 0.9mg, about 1mg, about 2mg, about 3mg, about 4mg, about 5mg, about 6mg, about 7mg, about 8mg, about 9mg, about 10mg, about 12mg, about 20mg, about 30mg, about 50mg, about 60mg, About 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300mg, about 330mg, about 380m
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 1-50:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 5-25:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 1-15:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 1-10:1.
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of about 5:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 15-30:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 20-30:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of about 25:1.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 1-50:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 5-25:1.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 1-15:1.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 1-10:1.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is approximately 5:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 15-30:1.
  • the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 20-30:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is approximately 25:1.
  • the anti-TIGIT antibody is antibody h10D8OFKF and the anti-CTLA-4 antibody is antibody 1.
  • the anti-TIGIT antibody is antibody h10D8OF and the anti-CTLA-4 antibody is antibody 1.
  • the formulations may be formulations suitable for injectable use including sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
  • Suitable carriers include physiological saline, bacteriostatic water or phosphate buffered saline (PBS), solvents or dispersion media of ethanol, polyols (eg, glycerol, propylene glycol, liquid polyethylene glycol, etc.), and suitable mixtures thereof.
  • the formulation contains at least 0.1% antibody or antigen-binding fragment.
  • a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 2 months, 5 months, 6 months, one year, or any combination thereof.
  • the dosing frequency ranges from once daily to once every 7 weeks.
  • the dosing frequency is once daily, three times per week, twice per week, once per week, once every 2 weeks, once every 3 weeks, every 4 weeks. Dosing once a week, once every 5 weeks, once every 6 weeks, or once every 7 weeks.
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment may be administered at the same or different frequencies.
  • the patient receives a single treatment with an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
  • the patient's symptoms are relieved after a single administration.
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered to the patient after the patient does not experience expected relief of symptoms after a single dose.
  • patients receive treatment until the condition resolves and treatment is no longer required.
  • the patient is administered an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment separately once per treatment cycle.
  • the patient is administered the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment multiple times per treatment cycle, such as 2, 3, 4, or 5 times.
  • the patient receives one treatment cycle.
  • a patient is treated with multiple treatment cycles (eg, 2, 3, or 4).
  • the anti-TIGIT antibody or antigen-binding fragment, anti-CTLA-4 antibody or antigen-binding fragment is administered by subcutaneous (s.c.) injection, intraperitoneal (i.p.) injection, parenteral injection, intraarterial injection, or intravenous (i.v.) injection. ) administration by injection or infusion.
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment can be administered in the same or different ways.
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered by intraperitoneal injection.
  • the anti-TIGIT antibody eg, antibody h10D8OF or h10D8OFKF
  • antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are separate administration units and administered in combination.
  • the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment can be administered at different times.
  • the anti-TIGIT antibody or antigen-binding fragment is administered prior to administration of the anti-CTLA-4 antibody or antigen-binding fragment.
  • the anti-TIGIT antibody or antigen-binding fragment is administered after administration of the anti-CTLA-4 antibody or antigen-binding fragment.
  • the anti-TIGIT antibody or antigen-binding fragment is administered simultaneously with the anti-CTLA-4 antibody or antigen-binding fragment.
  • An anti-TIGIT antibody eg, antibody h10D8OF or h10D8OFKF
  • an anti-CTLA-4 antibody or antigen-binding fragment are formulated into a pharmaceutical composition and administered to the patient in a form suitable for the chosen route of administration, for example, enterally External, intravenous (iv), intramuscular, topical or subcutaneous routes.
  • the anti-TIGIT antibody eg, antibody h10D8OF or h10D8OFKF
  • antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment simultaneously form a combined dosage unit for combined administration.
  • an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment are combined into a pharmaceutical composition and administered to the patient in a form suitable for the chosen route of administration, such as enterally. External, intravenous (iv), intramuscular, topical or subcutaneous routes.
  • anti-TIGIT antibodies or antigen-binding fragments (or formulations), anti-CTLA-4 antibodies or antigen-binding fragments (or formulations) can be used in combination with other treatments to treat tumors, such as chemotherapy, radiation therapy, and surgical treatment. wait.
  • tumors include, but are not limited to, benign tumors, cancer.
  • tumors include, but are not limited to, hematological cancers, solid tumors.
  • blood cancers include, but are not limited to, leukemias, lymphomas, and myeloma.
  • leukemias include acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and myeloid proliferative diseases/neoplasms (MPDS ).
  • ALL acute lymphoblastic leukemia
  • AML acute myelogenous leukemia
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • MPDS myeloid proliferative diseases/neoplasms
  • lymphomas include Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, and follicular lymphoma (small cell and large cell).
  • myeloma includes multiple myeloma (MM), giant cell myeloma, heavy chain myeloma, and light chain or Bence-Jones myeloma.
  • solid tumors include breast cancer, pancreatic cancer, prostate cancer, melanoma, head and neck cancer, liver cancer, renal cancer, squamous cell carcinoma, esophageal squamous cell carcinoma, lung cancer, non-small cell lung cancer, cervical cancer, Esophageal cancer, endometrial cancer, ovarian cancer, colon cancer, colorectal cancer, urothelial cancer, bladder cancer, brain cancer.
  • the tumor is a pathologically confirmed locally advanced or metastatic malignant solid tumor for which there is no effective treatment.
  • the combination of an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment compared to treatment with an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment alone can improve tumor suppression and/or prolong survival.
  • Figure 1 is the mouse tumor growth curve in Example 2.
  • Figure 2 is the mouse survival curve in Example 2.
  • Figure 3 is the mouse tumor growth curve in Example 3.
  • Figure 4 is the mouse survival curve in Example 3.
  • an entity refers to one or more such entities, e.g. "an antibody” should be understood to mean one or more antibodies, therefore, the term “a” (or “an” ), “one or more” and “at least one” may be used interchangeably herein.
  • polypeptide is intended to encompass the singular “polypeptide” as well as the plural “polypeptide” and refers to a molecule composed of amino acid monomers linearly linked 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 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 two or more amino acid chains, and the term “polypeptide” may Used instead of or 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-natural Amino acid modifications that occur.
  • a polypeptide may be derived from natural biological sources or produced by recombinant techniques, but it does not have to be translated from a specified nucleic acid sequence and 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 may 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 the "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 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) with similar chemical properties (eg, charge or hydrophobicity). Generally speaking, conservative amino acid substitutions are unlikely to materially alter the functional properties of the protein. Examples of 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.
  • Amide-containing side chains asparagine and glutamine
  • Aromatic side chains phenylalanine, tyrosine and tryptophan
  • Basic side chains lysine, Arginine and histidine
  • Acidic side chains aspartic acid and glutamic acid.
  • the number of amino acids of "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 between any two of these values (inclusive of the endpoints) or any one thereof What value.
  • the number of amino acids of "conservative amino acid substitutions of heavy or light chain” may 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 that is said to "encode” a polypeptide that, in its native state or when manipulated by methods well known to those skilled in the art, is transcribed and/or Or translation can produce the polypeptide and/or fragments thereof.
  • the 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, and antigen-binding fragments. (eg Fab, Fab', F(ab') 2 , scFv).
  • recombinant refers to a polypeptide or polynucleotide and means a form of the polypeptide or polynucleotide that does not occur in nature, and non-limiting examples may be combined to produce polynucleotides that do not normally exist or Peptides.
  • Homology 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. When a position in the compared sequences is occupied by the same base or amino acid, the molecules are homologous at that position. 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 means about 80% identity, about 81% identity, about 82% identity, about 83% identity, about 85% identity, about 86% identity, about 87% identity, About 88% identical, about 90% identical, about 91% identical, about 92% identical, about 94% identical, about 95% identical, about 98% identical, about 99% identical, or these The range between any two values in a numeric value, including endpoints, or any value within it.
  • a nucleic acid or polynucleotide sequence is "identical” or “sequence identical” to another sequence by a certain percentage (eg, 90%, 95%, 98% or 99%). When sequences are aligned, this percentage of bases (or amino acids) in the two sequences being compared are identical.
  • the alignment percent identity or sequence identity can be determined using visual inspection or software programs known in the art, such as those described in Ausubel et al. eds. (2007), Current Protocols in Molecular Biology. It is preferred to use the default parameters for comparison.
  • Biologically equivalent polynucleotides are polynucleotides that share the percentage identity specified above and encode a polypeptide with the same or similar biological activity.
  • Antibody and antigen-binding fragment refer to polypeptides or polypeptide complexes that specifically recognize and bind to antigens. Antibodies can be complete antibodies, 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 of binding to an antigen.
  • anti- Body and antigen-binding fragments include, but are not limited to, complementarity determining regions (CDRs) of heavy or light chains or their ligand-binding portions, heavy chain variable regions (VH), light chain variable regions (VL), heavy chain constant regions (CH), light chain constant region (CL), framework region (FR) or any part thereof, or at least part of a binding protein.
  • CDR region includes the CDR region of the light chain (L CDR1-3) and the CDR region of the heavy chain (HCDR1-3).
  • antibodies includes a wide variety of biochemically distinguishable polypeptides. Those skilled in the art will understand that classes of heavy chains include gamma, mu, alpha, delta or epsilon (gamma, mu, alpha, delta, epsilon), of which there are also subclasses (eg ⁇ 1- ⁇ 4). The nature of this chain determines the "class" of the antibody: IgG, IgM, IgA, IgG, or IgE. Immunoglobulin subclasses (isotypes) such as IgG1, IgG2, IgG3, IgG4, IgG5, etc. are well characterized and the functional specificities conferred are known. All immunoglobulin species are within the scope of the invention. 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 combined with a kappa or lambda light chain.
  • ⁇ kappa
  • ⁇ lambda
  • the amino acid sequence extends from the N-terminus at the fork 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 ⁇ .
  • the light chain variable region (VL) and heavy chain variable region (VH) determine antigen recognition and specificity.
  • the light chain constant region (CL) and heavy chain constant region (CH) confer important biological properties, such as secretion, transplacental movement, Fc receptor binding, complement binding, etc. By convention, the numbering of constant regions increases as they become farther away from the antibody's antigen-binding site, or amino terminus.
  • the N-terminal part is the variable region and the C-terminal part is the constant region; for example, the CH3 and CL domains of IgG1 actually contain the carboxyl termini of the heavy and light chains respectively.
  • CDR complementarity determining region
  • CDRs defined according to Kabat and Chothia include overlapping 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 numbering comprising 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 CDR contains 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 without relying on experimental data other than the sequence itself.
  • “Kabat numbering” refers to the numbering system proposed by Kabat et al., USDept. of Health and Human Services in "Sequence of Proteins of Immunological Interest” (1983).
  • Antibodies can also use the EU or Chothia numbering system.
  • Treatment means therapeutic treatment and preventive or preventative measures designed to prevent, slow down, ameliorate and halt adverse physiological changes or disorders such as the progression of a disease, including but not limited to the following whether detectable or undetectable
  • the results include alleviation of symptoms, reduction in disease severity, stabilization of disease status (i.e. no worsening), delay or slowdown of disease progression, improvement or alleviation of disease status, reduction or disappearance (whether partial or complete), prolongation and Expected survival without treatment, etc.
  • Patients in need of treatment include patients who already have a condition or disorder, are susceptible to a condition or disorder, or are in need of prevention of a condition or disorder from which an antibody or composition disclosed herein can be or is expected to result from administration of an antibody or composition disclosed herein for detection, Patients who benefit from diagnostic procedures and/or treatments.
  • Patient refers to any mammal for whom diagnosis, prognosis or treatment is required, including humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, etc.
  • Effective amount refers to an amount of an active compound or agent that causes a biological or medical response in a tissue, system, animal, individual, or human.
  • in need means that a patient has been identified as needing a particular method or treatment. In some embodiments, identification can be by any diagnostic means.
  • DNA encoding the antibody can be designed and synthesized according to the antibody amino acid sequence described herein according to conventional methods, placed into an expression vector, and then transfected into host cells, and the transfected host cells are cultured in culture medium to produce monoclonal antibodies.
  • an expression antibody 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 through the early and late promoters of SV40, the long terminal repeat sequences from retroviruses such as RSV, HTLV1, HIVI, and the early promoter of cytomegalovirus, and other cellular promoters such as muscle can also be used.
  • Kinesin 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 host cells include HEK293 cells, Cos1 cells, Cos7 cells, CV1 cells, mouse L cells and CHO cells.
  • the antibody heavy chain and light chain DNA sequences are cloned into expression vectors, then transferred into host cells, cultured and purified to obtain antibodies.
  • the amino acid sequences of antibody h10D8OF, antibody h10D8OFKF, antibody 1, antibody Anti-mCTLA4, antibody Anti-mTIGIT and control antibody IgG1 are shown in Table 1-5. Among them, the amino acid sequences of antibodies h10D8OF and h10D8OFKF are the same.
  • the host cells expressing antibody h10D8OFKF and antibody 1 are CHO-K1 cells with ⁇ -(1,6)-fucosyltransferase gene (Fut-8) knockout, and the resulting fucosylation level of antibody h10D8OFKF Approximately 0%, Antibody 1 fucosylation level is approximately 0%, total high mannose glycoforms ⁇ 2%, and total sialylated glycoforms ⁇ 1%.
  • the host cells expressing antibody h10D8OF are CHO-K1 cells.
  • the host cells expressing antibody Anti-mCTLA4, antibody Anti-mTIGIT and control antibody IgG1 are HEK293F cells.
  • mice were subcutaneously inoculated with CT26 colon cancer cells (inoculation number: 1 ⁇ 10 6 /mouse).
  • the day of grouping was recorded as day 0, which is the start.
  • the first administration, administration mode (administration frequency, cycle, route) and specific group administration regimen are shown in Table 6.
  • the mouse body weight and tumor volume were recorded twice a week, and the tumor inhibition rate TGI% and p value were calculated. The results were expressed as mean ⁇ standard error. Mice were euthanized when their tumor volume was greater than 3000 mm 3 .
  • mice tumor growth curve is shown in Figure 1, and the mouse survival curve is shown in Figure 2.
  • both the single-drug group (Group 2 and Group 3) and the combined administration group (Group 4) inhibited tumor growth to a certain extent, and no significant difference was shown between the groups 14 days after administration.
  • mice whose tumor volume reached the euthanasia standard were all euthanized. From the survival curve of the mice, it can be seen that the survival time of the mice in the combined administration group (group 4) was significantly longer than that in the negative control group ( Group 1) and single drug group (Group 2 and Group 3).
  • Example 3 Efficacy test of combined administration of antibody h10D8OFKF and antibody 1
  • mice were subcutaneously inoculated with CT26 cells at an inoculation number of 1 ⁇ 10 6 /mouse.
  • the average tumor volume of the mice was 88.33 mm 3 , they were divided into groups and administered drugs.
  • the day of grouping was recorded as day 0, that is, the first administration was started.
  • the administration method administration frequency, cycle, route
  • the specific group administration scheme are shown in Table 7.
  • the mouse body weight and tumor volume were recorded twice a week, and the tumor inhibition rate TGI% and p value were calculated. The results were expressed as mean ⁇ standard error. Mice were euthanized when their tumor volume was greater than 3000 mm 3 .
  • the mouse tumor growth curve is shown in Figure 3, and the mouse survival curve is shown in Figure 4.
  • the results of the study showed that compared with the control antibody IgG1, the test antibody 1, antibody h10D8OFKF, and the combination of antibody 1 and antibody h10D8OFKF all showed significant tumor inhibitory effects (see Figure 3) and could significantly extend the survival time of mice. (See Figure 4).
  • the group administered antibody 1 in combination with antibody h10D8OFKF group Compared with the group administered with antibody h10D8OFKF alone (group 3), the group administered antibody 1 in combination with antibody h10D8OFKF (group 4) showed significant tumor inhibition (*, p ⁇ 0.05) and could significantly prolong the survival of mice ( **, p ⁇ 0.01).

Abstract

The present invention relates to a combination therapy, comprising an anti-TIGIT antibody or an antigen-binding fragment thereof and an anti-CTLA-4 antibody or an antigen-binding fragment thereof. The present invention also relates to combined use of an anti-TIGIT antibody and an anti-CTLA-4 antibody in treating a tumor.

Description

抗TIGIT抗体和抗CTLA-4抗体联合用于***中的应用Use of anti-TIGIT antibodies and anti-CTLA-4 antibodies in combination to treat tumors 技术领域Technical field
本发明属于生物医药领域，具体涉及抗TIGIT抗体和抗CTLA-4抗体联合用于***中的应用。The invention belongs to the field of biomedicine, and specifically relates to the use of anti-TIGIT antibodies and anti-CTLA-4 antibodies in combination to treat tumors.
背景技术Background technique
TIGIT(T cell immunoreceptor with Ig and ITIM domains)是一种具有免疫球蛋白(Ig)和酪氨酸抑制剂基序(ITIM)结构域的T细胞免疫受体，主要表达于激活的T细胞和NK细胞上。TIGIT结构显示包含一个细胞外免疫球蛋白结构域，一个I型跨膜区和两个ITIM基序。TIGIT是共同刺激网络的一部分，这个共刺激网络主要由T细胞上的激活性受体CD226和抑制性受体TIGIT，以及在APC、肿瘤细胞、感染的细胞表面表达的配体CD155(也称为PVR，一种在人类中被PVR基因编码的脊髓灰质炎病毒受体蛋白质)和CD112组成。TIGIT与PVR或CD112结合后会引起TIGIT胞质内Tyr225被磷酸化，TIGIT和细胞自适应生长因子受体结合蛋白2(GRB2)进行结合。GRB2可以招募SHIP1抑制磷脂酰肌醇三激酶(PI3K)和促***原活化蛋白激酶(MAPK)信号。除此之外，磷酸化的TIGIT通过Beta抑制蛋白2(β-arrestin2)招募SHIP1和通过阻断TNF受体相关因子6(TRAF6)的自身泛素化并破坏核因子KB(NF-KB)激活，一系列的信号传导最终导致T细胞或NK细胞的功能受到抑制，细胞因子的产生受到抑制。PVR既是TIGIT的配体，又是CD226分子的配体。在和CD112或CD155结合之后，CD226的胞内结构域的Ser329和Tyr322被磷酸化；Ser329磷酸化促进蛋白激酶(PKC)的激活和CD226与淋巴细胞关联抗原1(LFA1)的相互结合。LFA1然后被用于TYN介导的Tyr322磷酸化和CD226介导的下游信号传导。一系列的信号传导最终导致T细胞或NK细胞的功能受到激活，促进细胞因子的产生。存在于T细胞或NK细胞表面的TIGIT分子与CD226分子之间也发生着相互作用，表现在TIGIT分子可以直接扰乱CD226形成正常的二聚体，从而破坏CD226的正常生理功能。TIGIT和CD226如同天平的两端，通过PVR这个支点，通过共刺激和共抑制信号的传导巧妙地调节着机体的免疫功能。TIGIT (T cell immunoreceptor with Ig and ITIM domains) is a T cell immunoreceptor with immunoglobulin (Ig) and tyrosine inhibitor motif (ITIM) domains, mainly expressed on activated T cells and NK on cells. The TIGIT structure is shown to contain an extracellular immunoglobulin domain, a type I transmembrane region and two ITIM motifs. TIGIT is part of a costimulatory network, which mainly consists of the activating receptor CD226 and the inhibitory receptor TIGIT on T cells, as well as the ligand CD155 (also known as CD155) expressed on the surface of APCs, tumor cells, and infected cells. PVR, a poliovirus receptor protein encoded by the PVR gene in humans) and CD112. The binding of TIGIT to PVR or CD112 will cause the phosphorylation of Tyr225 in the cytoplasm of TIGIT, and TIGIT will bind to the cell adaptive growth factor receptor binding protein 2 (GRB2). GRB2 can recruit SHIP1 to inhibit phosphatidylinositol trikinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling. In addition, phosphorylated TIGIT recruits SHIP1 through beta-arrestin2 (β-arrestin2) and blocks the autoubiquitination of TNF receptor-associated factor 6 (TRAF6) and disrupts nuclear factor KB (NF-KB) activation. , a series of signal transduction ultimately leads to the inhibition of T cell or NK cell function and the inhibition of cytokine production. PVR is both a ligand for TIGIT and a ligand for CD226 molecules. After binding to CD112 or CD155, Ser329 and Tyr322 of the intracellular domain of CD226 are phosphorylated; Ser329 phosphorylation promotes the activation of protein kinase (PKC) and the mutual binding of CD226 to lymphocyte-associated antigen 1 (LFA1). LFA1 is then used for TYN-mediated Tyr322 phosphorylation and CD226-mediated downstream signaling. A series of signal transduction ultimately leads to the activation of T cell or NK cell function and promotes the production of cytokines. There is also an interaction between TIGIT molecules present on the surface of T cells or NK cells and CD226 molecules, which means that TIGIT molecules can directly disrupt CD226 to form normal dimers, thereby destroying the normal physiological functions of CD226. TIGIT and CD226 are like two ends of a scale. Through the fulcrum of PVR, they subtly regulate the body's immune function through the transmission of co-stimulatory and co-inhibitory signals.
细胞毒性T淋巴细胞抗原-4(CTLA-4)是一种影响T细胞功能的关键抑制性受体，在免疫应答的启动阶段发挥关键作用(Scalapino KJ1,Daikh DI.(2008),Immunol Rev.223:143-155)。当抗原被MHC-I类或II类抗原提呈细胞运输到T细胞受体(TCR)时，TCR上的信号就会放大，并被共刺激分子抵消。T细胞上的CD28与抗原提呈细胞上的B7-1(CD80)和B7-2(CD86)结合后，会发出一种需要T细胞全激活的信号。 这种CD28/B7的结合导致白介素2及其他刺激性的细胞因子产生的增加，提高代谢，促进细胞周期进程，上调细胞存活基因，最终使T细胞得到增殖和分化。当CD28结合并引起T细胞增殖后，CTLA-4被运送并表达在T细胞表面(Linsley PS,et al.(1996),Immunity.4(6):535-543)。通过TCR的激活信号越强烈，就越多CTLA-4运送并表达。当在细胞表面上时，CTLA-4的抑制信号就被传播(Egen JG,Allison JP.(2002),Immunity.16(1):23-25)。与CD28相比，CTLA-4与B7有更高的亲和力，并能阻碍更进一步的共激活(Krummel MF,Allison JP.(1995),J Exp Med.182(2):458-465)。并且，CTLA-4表达的细胞能通过内吞作用捕获和降解B7-1和B7-2(Qureshi OS,et al.(2011),Science.332(6029):600-603)。Cytotoxic T lymphocyte antigen-4 (CTLA-4) is a key inhibitory receptor that affects T cell function and plays a key role in the initiation phase of immune response (Scalapino KJ1, Daikh DI. (2008), Immunol Rev. 223:143-155). When antigen is transported to the T cell receptor (TCR) by MHC class I or class II antigen-presenting cells, the signal on the TCR is amplified and counteracted by costimulatory molecules. After CD28 on T cells binds to B7-1 (CD80) and B7-2 (CD86) on antigen-presenting cells, it sends a signal that requires full activation of T cells. This CD28/B7 combination leads to an increase in the production of interleukin 2 and other stimulatory cytokines, increases metabolism, promotes cell cycle progression, upregulates cell survival genes, and ultimately enables T cells to proliferate and differentiate. When CD28 binds and causes T cell proliferation, CTLA-4 is transported and expressed on the surface of T cells (Linsley PS, et al. (1996), Immunity. 4(6):535-543). The stronger the activation signal through the TCR, the more CTLA-4 is transported and expressed. While on the cell surface, CTLA-4's inhibitory signal is propagated (Egen JG, Allison JP. (2002), Immunity. 16(1):23-25). CTLA-4 has a higher affinity for B7 than CD28 and blocks further coactivation (Krummel MF, Allison JP. (1995), J Exp Med. 182(2):458-465). Moreover, cells expressing CTLA-4 can capture and degrade B7-1 and B7-2 through endocytosis (Qureshi OS, et al. (2011), Science. 332 (6029): 600-603).
鉴于免疫检查点的重要性，亟需开发新的联合疗法。Given the importance of immune checkpoints, there is an urgent need to develop new combination therapies.
发明内容Contents of the invention
本发明的目的是提供一种联合用药物及其用于***中的应用。The object of the present invention is to provide a combined drug and its application in treating tumors.
在一些实施方案中，所述联合用药物包括抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段。在一些实施方案中，所述联合用药物包括治疗有效量的抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段。In some embodiments, the combination includes an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the combination includes a therapeutically effective amount of an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
另一方面，本发明提供***的药物组合，包括抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段。On the other hand, the present invention provides a pharmaceutical combination for treating tumors, including an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
另一方面，本发明提供了抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段联合用于***。在一些实施方案中，本发明提供了一种***的方法，其包括向有需要的患者施用有效量的抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段。In another aspect, the invention provides anti-TIGIT antibodies or antigen-binding fragments and anti-CTLA-4 antibodies or antigen-binding fragments in combination for the treatment of tumors. In some embodiments, the invention provides a method of treating tumors comprising administering to a patient in need thereof an effective amount of an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment.
另一方面，本发明提供了抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段在制备与抗CTLA-4抗体或抗原结合片段联合用于***的药物中的应用。In another aspect, the invention provides the use of an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or an antigen-binding fragment in the preparation of a medicament for treating tumors in combination with an anti-CTLA-4 antibody or antigen-binding fragment.
另一方面，本发明提供了抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段和抗CTLA-4抗体或抗原结合片段在制备用于***的药物中的应用。In another aspect, the invention provides the use of an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment in the preparation of a medicament for treating tumors.
另一方面，本发明提供了抗CTLA-4抗体或抗原结合片段在制备与抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段联合用于***的药物中的应用。In another aspect, the invention provides use of an anti-CTLA-4 antibody or antigen-binding fragment in the preparation of a medicament for treating tumors in combination with an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment.
另一方面，本发明提供了抗CTLA-4抗体或抗原结合片段和抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段在制备用于***的药物中的应用。In another aspect, the invention provides the use of an anti-CTLA-4 antibody or antigen-binding fragment and an anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment in the preparation of a medicament for treating tumors.
另一方面，本发明提供了抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段和抗CTLA-4抗体或抗原结合片段在制备用于***的药物组合物中 的应用。In another aspect, the invention provides anti-TIGIT antibodies (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragments and anti-CTLA-4 antibodies or antigen-binding fragments in the preparation of pharmaceutical compositions for treating tumors. Applications.
另一方面，本发明提供了一种试剂盒，其包含抗TIGIT抗体和包装插页，该包装插页包含关于与抗CTLA-4抗体组合使用所述抗TIGIT抗体***的说明书。In another aspect, the invention provides a kit comprising an anti-TIGIT antibody and a package insert containing instructions for using the anti-TIGIT antibody in combination with an anti-CTLA-4 antibody to treat tumors.
另一方面，本发明提供了一种试剂盒，其包含抗CTLA-4抗体和包装插页，该包装插页包含关于与抗TIGIT抗体组合使用所述抗CTLA-4抗体***的说明书。In another aspect, the invention provides a kit comprising an anti-CTLA-4 antibody and a package insert containing instructions for using the anti-CTLA-4 antibody in combination with an anti-TIGIT antibody to treat tumors.
另一方面，本发明提供了一种试剂盒，其包含抗TIGIT抗体和抗CTLA-4抗体的药剂，和包装插页，该包装插页包含关于使用所述抗TIGIT抗体和抗CTLA-4抗体的药剂***的说明书。In another aspect, the present invention provides a kit comprising a medicament of an anti-TIGIT antibody and an anti-CTLA-4 antibody, and a package insert containing instructions for using the medicament of an anti-TIGIT antibody and an anti-CTLA-4 antibody. Instructions for treating tumors.
另一方面，本发明提供了一种试剂盒，其包含抗TIGIT抗体的药剂和包装插页，该包装插页包含关于与抗CTLA-4抗体组合使用所述抗TIGIT抗体的药剂***的说明书。In another aspect, the present invention provides a kit comprising a medicament of an anti-TIGIT antibody and a package insert containing instructions for using the medicament of an anti-TIGIT antibody in combination with an anti-CTLA-4 antibody to treat tumors.
另一方面，本发明提供了一种试剂盒，其包含抗CTLA-4抗体的药剂和包装插页，该包装插页包含关于与抗TIGIT抗体组合使用所述抗CTLA-4抗体的药剂***的说明书。In another aspect, the present invention provides a kit comprising a medicament of an anti-CTLA-4 antibody and a package insert containing instructions for using the medicament of an anti-CTLA-4 antibody in combination with an anti-TIGIT antibody to treat tumors. .
另一方面，本发明提供了一种试剂盒，包含抗TIGIT抗体或抗原结合片段(或制剂)、抗CTLA-4抗体或抗原结合片段(或制剂)和用于指导有需要患者给药抗TIGIT抗体或抗原结合片段(或制剂)和抗CTLA-4抗体或抗原结合片段(或制剂)的说明书。On the other hand, the present invention provides a kit comprising an anti-TIGIT antibody or antigen-binding fragment (or preparation), an anti-CTLA-4 antibody or antigen-binding fragment (or preparation) and a method for instructing patients in need to administer anti-TIGIT Instructions for antibodies or antigen-binding fragments (or formulations) and anti-CTLA-4 antibodies or antigen-binding fragments (or formulations).
在一些实施方案中，所述抗TIGIT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-TIGIT 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 SEQ ID NO: 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TIGIT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-TIGIT 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TIGIT抗体或抗原结合片段包含重链可变区和轻链可变区。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region.
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:7所示的氨基酸序列，或与SEQ ID NO:7所示序列相比具有至少80％同一性的氨基酸序列，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。In some embodiments, the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:7, or is at least 80% identical to the sequence set forth in SEQ ID NO:7. A unique amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:7.
在一些实施方案中，所述抗TIGIT抗体或抗原结合片段的轻链可变区包含SEQ ID NO:8所示的氨基酸序列，或与SEQ ID NO:8所示序列相比具有至少80％同一性的氨基酸序列，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。 In some embodiments, the light chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:8, or is at least 80% identical to the sequence set forth in SEQ ID NO:8. A conservative amino acid sequence, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:8.
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:7所示的氨基酸序列，或与SEQ ID NO:7所示序列相比具有至少80％同一性的氨基酸序列，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；所述抗TIGIT抗体或抗原结合片段的轻链可变区包含SEQ ID NO:8所示的氨基酸序列，或与SEQ ID NO:8所示序列相比具有至少80％同一性的氨基酸序列，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。In some embodiments, the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:7, or is at least 80% identical to the sequence set forth in SEQ ID NO:7. A conservative amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared with the sequence shown in SEQ ID NO:7; the light chain variable region of the anti-TIGIT antibody or antigen-binding fragment includes SEQ ID NO:8 The amino acid sequence shown, or an amino acid sequence having at least 80% identity compared to the sequence shown in SEQ ID NO: 8, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 8 sequence.
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:7所示的氨基酸序列，所述抗TIGIT抗体或抗原结合片段的轻链可变区包含SEQ ID NO:8所示的氨基酸序列。In some embodiments, the heavy chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO:7, and the light chain variable region of the anti-TIGIT antibody or antigen-binding fragment comprises SEQ ID NO:7 The amino acid sequence shown in ID NO:8.
在一些实施方案中，所述抗TIGIT抗体的重链包含SEQ ID NO:9所示的氨基酸序列，或与SEQ ID NO:9所示序列相比具有至少80％同一性的氨基酸序列，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。In some embodiments, the heavy chain of the anti-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 9, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 9, or is identical to the sequence set forth in SEQ ID NO: 9. The sequence shown in SEQ ID NO:9 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
在一些实施方案中，所述抗TIGIT抗体的轻链包含SEQ ID NO:10所示的氨基酸序列，或与SEQ ID NO:10所示序列相比具有至少80％同一性的氨基酸序列，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。In some embodiments, the light chain of the anti-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 10, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 10, or is identical to the sequence set forth in SEQ ID NO: 10. The sequence shown in SEQ ID NO:10 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
在一些实施方案中，所述抗TIGIT抗体的重链包含SEQ ID NO:9所示的氨基酸序列，或与SEQ ID NO:9所示序列相比具有至少80％同一性的氨基酸序列，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；所述抗TIGIT抗体的轻链包含SEQ ID NO:10所示的氨基酸序列，或与SEQ ID NO:10所示序列相比具有至少80％同一性的氨基酸序列，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。In some embodiments, the heavy chain of the anti-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 9, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 9, or is identical to the sequence set forth in SEQ ID NO: 9. The sequence shown in SEQ ID NO:9 is an amino acid sequence with one or more conservative amino acid substitutions; the light chain of the anti-TIGIT antibody includes the amino acid sequence shown in SEQ ID NO:10, or is identical to the amino acid sequence shown in SEQ ID NO:10 An amino acid sequence that has at least 80% identity compared to the sequence shown in SEQ ID NO: 10, or an amino acid sequence that has one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 10.
在一些实施方案中，所述抗TIGIT抗体为抗体h10D8OF或h10D8OFKF，其重链包含如SEQ ID NO:9所示的氨基酸序列，其轻链包含如SEQ ID NO:10所示的氨基酸序列。在一些实施方案中，抗体h10D8OF或h10D8OFKF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 an antibody h10D8OF or h10D8OFKF, the heavy chain of which comprises the amino acid sequence shown in SEQ ID NO:9, and the light chain of which comprises the amino acid sequence of SEQ ID NO:10. In some embodiments, antibody h10D8OF or h10D8OFKF contains two sequence-identical heavy chains and two sequence-identical light chains.
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％、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 anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment has a fucosylation level of 0-10%. In some embodiments, the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment has a fucosylation level of 0-5%. In some embodiments, the anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment has a fucosylation level 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 The range between (inclusive) or any value therein. 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 (eg, antibody h10D8OFKF) or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段至少包含SEQ ID NO:11所示的HCDR1、SEQ ID NO:12所示的HCDR2、SEQ ID NO:13所示的HCDR3、SEQ ID NO:14所示的LCDR1、SEQ ID NO:15所示的LCDR2、SEQ ID NO:16所示的LCDR3中一个或多个。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment at least comprises HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, HCDR3 shown in SEQ ID NO: 13, SEQ One or more of LCDR1 shown in ID NO:14, LCDR2 shown in SEQ ID NO:15, and LCDR3 shown in SEQ ID NO:16.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段包含SEQ ID NO:11所示的HCDR1、SEQ ID NO:12所示的HCDR2、SEQ ID NO:13所示的HCDR3、SEQ ID NO:14所示的LCDR1、SEQ ID NO:15所示的LCDR2和SEQ ID NO:16所示的LCDR3。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, HCDR3 shown in SEQ ID NO: 13, SEQ ID LCDR1 shown in NO:14, LCDR2 shown in SEQ ID NO:15 and LCDR3 shown in SEQ ID NO:16.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段包含重链可变区和轻链可变区。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的重链可变区包含SEQ ID NO:17所示的氨基酸序列，或与SEQ ID NO:17所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:17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-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO: 17, or has an amino acid sequence of at least 80% compared to the sequence set forth in SEQ ID NO: 17. % identical amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:17.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的轻链可变区包含SEQ ID NO:18所示的氨基酸序列，或与SEQ ID NO:18所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:18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 light chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO: 18, or has an amino acid sequence of at least 80% compared to the sequence set forth in SEQ ID NO: 18. % identity of the amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:18.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的重链可变区包含SEQ ID NO:17所示的氨基酸序列，或与SEQ ID NO:17所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:17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；所述抗CTLA-4抗体或抗原结合片段的轻链可变区包含SEQ ID NO:18所示的氨基酸序列，或与SEQ ID NO:18所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:18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-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence set forth in SEQ ID NO: 17, or has an amino acid sequence of at least 80% compared to the sequence set forth in SEQ ID NO: 17. % identical amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 17; the light chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment includes SEQ The amino acid sequence shown in ID NO:18, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:18, or one or more conserved amino acid sequences compared with the sequence shown in SEQ ID NO:18 Amino acid sequence of amino acid substitutions.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段的重链可变区包含SEQ ID NO:17所示的氨基酸序列，所述抗CTLA-4抗体或抗原结合片段的轻链可变区包含SEQ ID NO:18所示的氨基酸序列。In some embodiments, the heavy chain variable region of the anti-CTLA-4 antibody or antigen-binding fragment comprises the amino acid sequence shown in SEQ ID NO: 17, and the light chain of the anti-CTLA-4 antibody or antigen-binding fragment can The variable region contains the amino acid sequence shown in SEQ ID NO:18.
在一些实施方案中，所述抗CTLA-4抗体的重链包含SEQ ID NO:19所示的氨基酸序列，或与SEQ ID NO:19所示序列相比具有至少80％同一性的氨基酸序列，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。In some embodiments, the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 19, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 19, Or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:19.
在一些实施方案中，所述抗CTLA-4抗体的轻链包含SEQ ID NO:20所示的氨基酸序列，或与SEQ ID NO:20所示序列相比具有至少80％同一性的氨基酸序列，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 light chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO:20, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO:20, Or an amino acid sequence with 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CTLA-4抗体的重链包含SEQ ID NO:19所示的氨基酸序列，或与SEQ ID NO:19所示序列相比具有至少80％同一性的氨基酸序列，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；所述抗CTLA-4抗体的轻链包含SEQ ID NO:20所示的氨基酸序列，或与SEQ ID NO:20所示序列相比具有至少80％同一性的氨基酸序列，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 of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 19, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 19, or with SEQ The sequence shown in ID NO: 19 is compared to the amino acid sequence having one or more conservative amino acid substitutions; the light chain of the anti-CTLA-4 antibody includes the amino acid sequence shown in SEQ ID NO: 20, or is the same as SEQ ID NO: 20 The sequence shown is an amino acid sequence that has at least 80% identity compared to the sequence shown in SEQ ID NO: 20, or an amino acid sequence that has 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CTLA-4抗体的重链包含如SEQ ID NO:19所示的氨基酸序列，所述抗CTLA-4抗体的轻链包含如SEQ ID NO:20所示的氨基酸序列。在一些实施方案中，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 heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 19, and the light chain of the anti-CTLA-4 antibody comprises the amino acid sequence set forth in SEQ ID NO: 20 sequence. In some embodiments, an anti-CTLA-4 antibody contains two sequence-identical heavy chains and two sequence-identical light chains.
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％、0.2％、约0.36％、约0.8％、约1.5％、约2.2％、约2.7％、约2.9％、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 a total amount of high-mannose glycoforms <5% and/or a total amount of sialylated glycoforms <3%. 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 of the endpoints) or any value therein. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a total amount of sialylated glycoforms of about 0.1%, 0.2%, about 0.36%, about 0.8%, about 1.5%, about 2.2%, about 2.7 %, about 2.9%, 2.99%, or a range between any two of these values (inclusive of the endpoints) 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 a total amount of high mannose glycoforms <2% and/or a total amount of sialylated glycoforms <1%.
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0.5％、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CTLA-4抗体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CTLA-4抗体或抗原结合片段具有增强的ADCC效应(antibody-dependent cell-mediated cytotoxicity)。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 anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level 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 between any two of these values Range (including endpoints) or any value therein. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment does not bind fucose. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has enhanced ADCC effect (antibody-dependent cell-mediated cytotoxicity).
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％，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，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％，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，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％，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，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％，高甘露糖糖型总量<3％和/或唾液酸化糖型总量<2％。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％，高甘露糖糖型总量<3％和/或唾液酸化糖型总量<2％。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％，高甘露糖糖型总量<3％和/或唾液酸化糖型总量<2％。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％，高甘 露糖糖型总量<2％和/或唾液酸化糖型总量<1％。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％，高甘露糖糖型总量<2％和/或唾液酸化糖型总量<1％。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％，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 a fucosylation level of 0-10%, a total high-mannose glycoform <5% and/or a total sialylated glycoform <5%. 3%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%, a total high-mannose glycoform <5% and/or a total sialylated glycoform < 3%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of about 0%, a total high-mannose glycoform <5% and/or a total sialylated glycoform <3 %. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-10%, a total high-mannose glycoform <3% and/or a total sialylated glycoform <3%. 2%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%, a total high-mannose glycoform <3% and/or a total sialylated glycoform <3%. 2%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of about 0%, a total high-mannose glycoform <3% and/or a total sialylated glycoform <2 %. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-10%, high glycerol The total amount of unglucose glycoforms is <2% and/or the total amount of sialylated glycoforms is <1%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of 0-5%, a total high-mannose glycoform <2% and/or a total sialylated glycoform <2%. 1%. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment has a fucosylation level of about 0%, a total amount of high-mannose glycoforms <2%, and/or a total amount of sialylated glycoforms <1 %.
在一些实施方案中，所述抗CTLA-4抗体或抗原结合片段为伊匹单抗(YervoyTM或其生物类似物)。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is ipilimumab (Yervoy ™ or a biosimilar thereof).
抗体可以通过基因工程在宿主细胞中表达，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。Antibodies can be expressed in host cells through genetic engineering and obtained through purification; purification can be performed using conventional methods, such as centrifuging the cell suspension and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as ProteinA affinity columns and ion exchange columns can be used to purify antibodies.
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(例如抗体h10D8OFKF)或抗原结合片段或者抗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表达，例如由α-(1,6)-岩藻糖基转移酶基因敲除的CHO-K1细胞表达。In some embodiments, a hypofucosylated or afucosylated anti-TIGIT antibody (eg, antibody h10D8OFKF) or antigen-binding fragment or an anti-CTLA-4 antibody or antigen-binding fragment is composed of α-(1,6) - Expression from a fucosyltransferase gene knockout cell line, such as expression from α-(1,6)-fucosyltransferase gene knockout CHO-K1 cells.
在一些实施方案中，本发明还提供了包含抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的药物组合物。在一些实施方案中，所述药物组合物为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TIGIT抗体或抗原结合片段和0.1％的抗CTLA-4抗体或抗原结合片段。抗体的百分比可以变化，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TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段可以为给药的有效量。In some embodiments, the invention also provides pharmaceutical compositions comprising an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the pharmaceutical composition is a pharmaceutical composition suitable for injection, such as a push-type pharmaceutical composition or an infusion (drip)-type pharmaceutical composition. In some embodiments, the pharmaceutical composition contains at least 0.1% anti-TIGIT antibody or antigen-binding fragment and 0.1% anti-CTLA-4 antibody or antigen-binding fragment. The percentage of antibody can vary and can be between about 2% and about 90% by weight of a given dosage form. The anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment in this therapeutically useful pharmaceutical composition can be administered in effective amounts.
另一方面，本发明还提供了上述药物组合物的制备方法：分别将本文所述的抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段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。在一些实施方案中，上述药物组合物的制备方法：将本文所述的抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段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。上述抗体或抗原结合片段与载体的混合方法是本领域通常已知的。On the other hand, the present invention also provides a method for preparing the above-mentioned pharmaceutical composition: respectively, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment described herein are combined with a pharmaceutically acceptable suitable for injection. Mix with carrier (such as water for injection, physiological saline, etc.). In some embodiments, the above pharmaceutical composition is prepared by: combining the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment described herein with a pharmaceutically acceptable carrier suitable for injection (e.g., injection Mix with water, saline, etc.). Methods for mixing the above-described antibodies or antigen-binding fragments with carriers are generally known in the art.
在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为1-50:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为5-25:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为1-15:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为1-10:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为约5:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为15-30:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为20-30:1。在一些实施方 案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为约25:1。In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-50:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 5-25:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-15:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-10:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment to the anti-CTLA-4 antibody or antigen-binding fragment is about 5:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 15-30:1. In some embodiments, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 20-30:1. In some implementations In this case, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is approximately 25:1.
在一些实施方案中，抗TIGIT抗体或抗原结合片段每次施用的剂量为约0.05mg至1200mg、约0.5mg至1000mg、约5mg至500mg、约5mg至100mg、约10mg至100mg、约10mg至50mg，或含此剂量抗TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体或抗原结合片段每次施用的剂量为约0.05mg、约0.1mg、约0.5mg、约1mg、约2mg、约3mg、约4mg、约5mg、约6mg、约7mg、约8mg、约9mg、约10mg、约12mg、约20mg、约30mg、约40mg、约50mg、约60mg、约80mg、约100mg、约120mg、约200mg、约250mg、约290mg、约300mg、约330mg、约380mg、约400mg、约480mg、约500mg、约580mg、约600mg、约800mg、约900mg、约1000mg、约1200mg，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TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体为抗体h10D8OF或h10D8OFKF。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 1200 mg, about 0.5 mg to 1000 mg, about 5 mg to 500 mg, about 5 mg to 100 mg, about 10 mg to 100 mg, about 10 mg to 50 mg. , or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7mg, about 8mg, about 9mg, about 10mg, about 12mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300mg, About 330 mg, about 380 mg, about 400 mg, about 480 mg, about 500 mg, about 580 mg, about 600 mg, about 800 mg, about 900 mg, about 1000 mg, about 1200 mg, or a range between any two of these values (inclusive of the endpoints), or Any of these values, or a formulation containing this dose of anti-TIGIT antibody or antigen-binding fragment. In some embodiments, the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF.
在一些实施方案中，抗TIGIT抗体或抗原结合片段每次施用的剂量为约0.01mg/kg至20mg/kg、约0.01mg/kg至2mg/kg、约0.01mg/kg至1mg/kg、约0.05mg/kg至1mg/kg、约0.1mg/kg至1mg/kg，或含此剂量抗TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体或抗原结合片段每次施用的剂量为约0.01mg/kg、约0.05mg/kg、约0.1mg/kg、约0.3mg/kg、约0.5mg/kg、约0.8mg/kg、约0.9mg/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12mg/kg、约14mg/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，或含此剂量抗TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体为抗体h10D8OF或h10D8OFKF。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg to 20 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg/kg, about 0.05 mg/kg to 1 mg/kg, about 0.1 mg/kg to 1 mg/kg, or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg, about 0.8mg/kg, about 0.9mg/kg, about 1mg/kg, about 2mg/kg, about 3mg/kg, about 4mg/kg, about 5mg/kg, about 6mg/kg, about 7mg/kg, about 8mg/kg , about 9 mg/kg, about 10 mg/kg, about 12 mg/kg, about 14 mg/kg, about 15 mg/kg, about 18 mg/kg, about 20 mg/kg, or a range between any two of these values (including endpoint) or any value therein, or a formulation containing such a dose of anti-TIGIT antibody or antigen-binding fragment. In some embodiments, the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF.
在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约0.05mg至1200mg、约0.5mg至1000mg、约5mg至500mg、约5mg至100mg、约10mg至100mg、约10mg至50mg，或含此剂量抗TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约0.05mg、约0.1mg、约0.5mg、约1mg、约2mg、约3mg、约4mg、约5mg、约6mg、约7mg、约8mg、约9mg、约10mg、约12mg、约20mg、约30mg、约40mg、约50mg、约60mg、约80mg、约100mg、约120mg、约200mg、约250mg、约290mg、约300mg、约330mg、约380mg、约400mg、约480mg、约500mg、约580mg、约600mg、约800mg、约900mg、约1000mg、约1200mg，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TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体为抗体h10D8OF或h10D8OFKF。 In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 1200 mg, about 0.5 mg to 1000 mg, about 5 mg to 500 mg, about 5 mg to 100 mg, about 10 mg to 100 mg, about 10 mg per treatment cycle. to 50 mg, or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg per treatment cycle. , about 7mg, about 8mg, about 9mg, about 10mg, about 12mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300 mg, about 330 mg, about 380 mg, about 400 mg, about 480 mg, about 500 mg, about 580 mg, about 600 mg, about 800 mg, about 900 mg, about 1000 mg, about 1200 mg, or a range between any two of these values, including endpoints ) or any value therein, or a formulation containing such a dose of anti-TIGIT antibody or antigen-binding fragment. In some embodiments, the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF.
在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约0.01mg/kg至20mg/kg、约0.01mg/kg至2mg/kg、约0.01mg/kg至1mg/kg、约0.05mg/kg至1mg/kg、约0.1mg/kg至1mg/kg，或含此剂量抗TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约0.01mg/kg、约0.05mg/kg、约0.1mg/kg、约0.3mg/kg、约0.5mg/kg、约0.8mg/kg、约0.9mg/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12mg/kg、约14mg/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，或含此剂量抗TIGIT抗体或抗原结合片段的制剂。在一些实施方案中，抗TIGIT抗体为抗体h10D8OF或h10D8OFKF。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered per treatment cycle at a dose of about 0.01 mg/kg to 20 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg/kg , about 0.05 mg/kg to 1 mg/kg, about 0.1 mg/kg to 1 mg/kg, or preparations containing anti-TIGIT antibodies or antigen-binding fragments at this dose. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.3 mg/kg, about 0.5 mg/kg per treatment cycle. , about 0.8mg/kg, about 0.9mg/kg, about 1mg/kg, about 2mg/kg, about 3mg/kg, about 4mg/kg, about 5mg/kg, about 6mg/kg, about 7mg/kg, about 8mg /kg, about 9 mg/kg, about 10 mg/kg, about 12 mg/kg, about 14 mg/kg, about 15 mg/kg, about 18 mg/kg, about 20 mg/kg, or a range between any two of these values (inclusive of the endpoints) or any value therein, or a formulation containing such a dose of anti-TIGIT antibody or antigen-binding fragment. In some embodiments, the anti-TIGIT antibody is antibody h10D8OF or h10D8OFKF.
在一些实施方案中，抗CTLA-4抗体或抗原结合片段每次施用的剂量为约0.05mg至600mg、约0.5mg至600mg、约1mg至100mg、约1mg至50mg、约1mg至10mg，或含此剂量抗CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体或抗原结合片段每次施用的剂量为约0.05mg、约0.1mg、约0.2mg、约0.3mg、约0.5mg、约0.8mg、约1mg、约2mg、约3mg、约4mg、约5mg、约6mg、约7mg、约8mg、约9mg、约10mg、约12mg、约20mg、约30mg、约50mg、约60mg、约80mg、约100mg、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CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体为抗体1。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 600 mg, about 0.5 mg to 600 mg, about 1 mg to 100 mg, about 1 mg to 50 mg, about 1 mg to 10 mg, or containing Preparation of anti-CTLA-4 antibodies or antigen-binding fragments at this dose. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.5 mg, about 0.8 mg, about 1 mg, about 2 mg per administration. , about 3mg, about 4mg, about 5mg, about 6mg, about 7mg, about 8mg, about 9mg, about 10mg, about 12mg, about 20mg, about 30mg, about 50mg, about 60mg, about 80mg, about 100mg, about 120mg, 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 (inclusive of the endpoints) or any value therein, or a formulation containing such a dose of anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the anti-CTLA-4 antibody is Antibody 1.
在一些实施方案中，抗CTLA-4抗体或抗原结合片段每次施用的剂量为约0.01mg/kg至10mg/kg、约0.01mg/kg至2mg/kg、约0.01mg/kg至1mg/kg、约0.02mg/kg至1mg/kg、约0.02mg/kg至0.5mg/kg、约0.02mg/kg至0.2mg/kg，或含此剂量抗CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体或抗原结合片段每次施用的剂量为约0.01mg/kg、约0.02mg/kg、约0.03mg/kg、约0.04mg/kg、约0.05mg/kg、约0.06mg/kg、约0.07mg/kg、约0.08mg/kg、约0.1mg/kg、约0.3mg/kg、约0.5mg/kg、约0.8mg/kg、约0.9mg/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，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CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体为抗体1。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg to 10 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg/kg per administration , about 0.02 mg/kg to 1 mg/kg, about 0.02 mg/kg to 0.5 mg/kg, about 0.02 mg/kg to 0.2 mg/kg, or preparations containing anti-CTLA-4 antibodies or antigen-binding fragments at this dose. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg/kg, about 0.04 mg/kg, about 0.05 mg/kg per administration. , about 0.06mg/kg, about 0.07mg/kg, about 0.08mg/kg, about 0.1mg/kg, about 0.3mg/kg, about 0.5mg/kg, about 0.8mg/kg, about 0.9mg/kg, 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, about 10 mg/kg, or The range between any two of these values (inclusive of the endpoints) or any value therein, or the formulation containing such doses of anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the anti-CTLA-4 antibody is Antibody 1.
在一些实施方案中，抗CTLA-4抗体或抗原结合片段每个治疗周期施用的剂量为约0.05mg至600mg、约0.5mg至600mg、约1mg至100mg、约1mg至50mg、约1mg至10mg，或含此剂量抗CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体或抗原结合片段每个治疗周期施用的剂量为约0.05mg、约0.1 mg、约0.2mg、约0.3mg、约0.5mg、约0.8mg、约1mg、约2mg、约3mg、约4mg、约5mg、约6mg、约7mg、约8mg、约9mg、约10mg、约12mg、约20mg、约30mg、约50mg、约60mg、约80mg、约100mg、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CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体为抗体1。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg to 600 mg, about 0.5 mg to 600 mg, about 1 mg to 100 mg, about 1 mg to 50 mg, about 1 mg to 10 mg per treatment cycle, or preparations containing anti-CTLA-4 antibodies or antigen-binding fragments at this dose. In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.05 mg, about 0.1 mg per treatment cycle mg, about 0.2mg, about 0.3mg, about 0.5mg, about 0.8mg, about 1mg, about 2mg, about 3mg, about 4mg, about 5mg, about 6mg, about 7mg, about 8mg, about 9mg, about 10mg, about 12mg , about 20mg, about 30mg, about 50mg, about 60mg, about 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300mg, about 330mg, about 380mg, about 400mg, about 434mg, about 480mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, or a range between any two of these values (inclusive of the endpoints) or any value therein, or a formulation containing such a dose of anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the anti-CTLA-4 antibody is Antibody 1.
在一些实施方案中，抗CTLA-4抗体或抗原结合片段每个治疗周期施用的剂量为约0.01mg/kg至10mg/kg、约0.01mg/kg至2mg/kg、约0.01mg/kg至1mg/kg、约0.02mg/kg至1mg/kg、约0.02mg/kg至0.5mg/kg、约0.02mg/kg至0.2mg/kg，或含此剂量抗CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体或抗原结合片段每个治疗周期施用的剂量为约0.01mg/kg、约0.02mg/kg、约0.03mg/kg、约0.04mg/kg、约0.05mg/kg、约0.06mg/kg、约0.07mg/kg、约0.08mg/kg、约0.1mg/kg、约0.3mg/kg、约0.5mg/kg、约0.8mg/kg、约0.9mg/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，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CTLA-4抗体或抗原结合片段的制剂。在一些实施方案中，抗CTLA-4抗体为抗体1。In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered at a dose of about 0.01 mg/kg to 10 mg/kg, about 0.01 mg/kg to 2 mg/kg, about 0.01 mg/kg to 1 mg per treatment cycle. /kg, about 0.02 mg/kg to 1 mg/kg, about 0.02 mg/kg to 0.5 mg/kg, about 0.02 mg/kg to 0.2 mg/kg, or preparations containing anti-CTLA-4 antibodies or antigen-binding fragments at this dose . In some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment is administered per treatment cycle at a dose of about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg/kg, about 0.04 mg/kg, about 0.05 mg /kg, about 0.06mg/kg, about 0.07mg/kg, about 0.08mg/kg, about 0.1mg/kg, about 0.3mg/kg, about 0.5mg/kg, about 0.8mg/kg, about 0.9mg/kg , about 1mg/kg, about 2mg/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 , or a range between any two of these values (inclusive of the endpoints) or any value therein, or a formulation containing such a dose of anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the anti-CTLA-4 antibody is Antibody 1.
在一些实施方案中，本发明提供了一种***的方法，所述方法包括：向有需要的患者施用约0.05mg至1200mg、约0.5mg至1000mg、约5mg至500mg、约5mg至100mg、约10mg至100mg、约10mg至50mg，比如约0.05mg、约0.08mg、约0.1mg、约0.2mg、约0.3mg、约0.4mg、约0.5mg、约0.6mg、约0.7mg、约0.8mg、约0.9mg、约1mg、约2mg、约3mg、约4mg、约5mg、约6mg、约7mg、约8mg、约9mg、约10mg、约12mg、约20mg、约30mg、约50mg、约60mg、约80mg、约100mg、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、约700mg、约800mg、约900mg、约1000mg、约1100mg、约1200mg的抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段，或含此剂量抗TIGIT抗体或抗原结合片段的制剂；还向有需要的患者施用约0.05mg至600mg、约0.5mg至600mg、约1mg至100mg、约1mg至50mg、约1mg至10mg，比如约0.05mg、约0.08mg、约0.1mg、约0.2mg、约0.3mg、约0.4mg、约0.5mg、约0.6mg、约0.7mg、约0.8mg、约0.9mg、约1mg、约2mg、约3mg、约4mg、约5mg、约6mg、约7mg、约8mg、约9mg、约10mg、约12mg、约20mg、约30mg、约50mg、约60mg、约80mg、约100mg、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的抗CTLA-4抗体或抗原结合片段，或含此剂量抗CTLA-4抗体或抗原结合片段的制剂。In some embodiments, the invention provides a method of treating tumors, the method comprising: administering to a patient in need thereof about 0.05 mg to 1200 mg, about 0.5 mg to 1000 mg, about 5 mg to 500 mg, about 5 mg to 100 mg, About 10 mg to 100 mg, about 10 mg to 50 mg, such as about 0.05 mg, about 0.08 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg , about 0.9mg, about 1mg, about 2mg, about 3mg, about 4mg, about 5mg, about 6mg, about 7mg, about 8mg, about 9mg, about 10mg, about 12mg, about 20mg, about 30mg, about 50mg, about 60mg, About 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300mg, about 330mg, about 380mg, about 400mg, about 434mg, about 480mg, about 500mg, about 567mg, about 580mg, about 600mg, about 700mg , about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg of anti-TIGIT antibodies (such as antibodies h10D8OF or h10D8OFKF) or antigen-binding fragments, or preparations containing such doses of anti-TIGIT antibodies or antigen-binding fragments; also to those in need The patient administers about 0.05 mg to 600 mg, about 0.5 mg to 600 mg, about 1 mg to 100 mg, about 1 mg to 50 mg, about 1 mg to 10 mg, such as about 0.05 mg, about 0.08 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, About 0.4mg, about 0.5mg, about 0.6mg, about 0.7mg, about 0.8mg, about 0.9mg, about 1mg, about 2mg, about 3mg, about 4mg, about 5mg, about 6mg, about 7mg, about 8mg, about 9mg , about 10mg, about 12mg, about 20mg, about 30mg, about 50mg, about 60mg, about 80mg, about 100mg, about 120mg, about 200mg, about 250mg, about 290mg, about 300mg, about 330mg, about 380mg, about 400mg, about 434mg, about 480mg, About 500 mg, about 567 mg, about 580 mg, about 600 mg of anti-CTLA-4 antibody or antigen-binding fragment, or a preparation containing such dosage of anti-CTLA-4 antibody or antigen-binding fragment.
在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为1-50:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为5-25:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为1-15:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为1-10:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为约5:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为15-30:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为20-30:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为约25:1。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 1-50:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 5-25:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 1-15:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 1-10:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of about 5:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 15-30:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of 20-30:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered at a mass ratio of about 25:1.
在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为1-50:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为5-25:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为1-15:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为1-10:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为约5:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为15-30:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为20-30:1。在一些实施方案中，抗TIGIT抗体或抗原结合片段每个治疗周期施用的剂量为约10mg、30mg、100mg、300mg、600mg、或900mg，且抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段给药的质量比为约25:1。 In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 1-50:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 5-25:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 1-15:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 1-10:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is approximately 5:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 15-30:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is 20-30:1. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of about 10 mg, 30 mg, 100 mg, 300 mg, 600 mg, or 900 mg per treatment cycle, and the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or The mass ratio of antigen-binding fragments administered is approximately 25:1.
在一些实施方案中，抗TIGIT抗体为抗体h10D8OFKF，抗CTLA-4抗体为抗体1。In some embodiments, the anti-TIGIT antibody is antibody h10D8OFKF and the anti-CTLA-4 antibody is antibody 1.
在一些实施方案中，抗TIGIT抗体为抗体h10D8OF，抗CTLA-4抗体为抗体1。In some embodiments, the anti-TIGIT antibody is antibody h10D8OF and the anti-CTLA-4 antibody is antibody 1.
制剂可以为适于注射用途的制剂包括无菌水性溶液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。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％的抗体或抗原结合片段。The formulations may be formulations suitable for injectable use including sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. Suitable carriers include physiological saline, bacteriostatic water or phosphate buffered saline (PBS), solvents or dispersion media of ethanol, polyols (eg, glycerol, propylene glycol, liquid polyethylene glycol, etc.), and suitable mixtures thereof. In some embodiments, the formulation contains at least 0.1% antibody or antigen-binding fragment.
在一些实施方案中，一个治疗周期为1周、2周、3周、4周、5周、6周、7周，2个月、5个月、6个月、一年，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，给药频率为每天给药一次至每7周给药一次。在一些实施方案中，给药频率为每天给药一次、每周给药三次、每周给药两次、每周给药一次、每2周给药一次、每3周给药一次、每4周给药一次、每5周给药一次、每6周给药一次或每7周给药一次。抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的给药频率可以相同或者不同。In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 2 months, 5 months, 6 months, one year, or any combination thereof. The range between any two values (inclusive) or any value within them. In some embodiments, the dosing frequency ranges from once daily to once every 7 weeks. In some embodiments, the dosing frequency is once daily, three times per week, twice per week, once per week, once every 2 weeks, once every 3 weeks, every 4 weeks. Dosing once a week, once every 5 weeks, once every 6 weeks, or once every 7 weeks. The anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment may be administered at the same or different frequencies.
在一些实施方案中，患者接受单次抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的治疗。在一些实施方案中，单次给药后，患者的症状得到缓解。在一些实施方案中，单次给药后，患者后的症状未得到预期缓解，再对患者给药抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段。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，患者每个治疗周期内分别给药一次抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段。在一些实施方案中，患者每个治疗周期内多次给药抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段，例如2次、3次、4次或5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，患者接受多个治疗周期(例如2个、3个或4个)治疗。In some embodiments, the patient receives a single treatment with an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the patient's symptoms are relieved after a single administration. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered to the patient after the patient does not experience expected relief of symptoms after a single dose. In some embodiments, patients receive treatment until the condition resolves and treatment is no longer required. In some embodiments, the patient is administered an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment separately once per treatment cycle. In some embodiments, the patient is administered the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment multiple times per treatment cycle, such as 2, 3, 4, or 5 times. In some embodiments, the patient receives one treatment cycle. In some embodiments, a patient is treated with multiple treatment cycles (eg, 2, 3, or 4).
在一些实施方案中，抗TIGIT抗体或抗原结合片段、抗CTLA-4抗体或抗原结合片段是通过皮下(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.)注射或输液等方式进行给药。抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段可以通过相同或者不同方式进行给药。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段是通过腹腔注射给药。In some embodiments, the anti-TIGIT antibody or antigen-binding fragment, anti-CTLA-4 antibody or antigen-binding fragment is administered by subcutaneous (s.c.) injection, intraperitoneal (i.p.) injection, parenteral injection, intraarterial injection, or intravenous (i.v.) injection. ) administration by injection or infusion. The anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment can be administered in the same or different ways. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are administered by intraperitoneal injection.
在一些实施方案中，抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段和抗CTLA-4抗体或抗原结合片段分别为独立的给药单元，联合用药。在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段可以在不同时间给药。在一些实施方案中，抗TIGIT抗体或抗原结合片段在施加抗CTLA-4抗体或抗原结合片段之前给药。在一些实施方案中，抗TIGIT抗体或抗原结合片段在施加抗CTLA-4抗体或抗原结合片段之后给药。在一些实施方案中，抗TIGIT抗体或抗原结合片段与抗CTLA-4抗体或抗原结合片段同时给药。在一些实施方案中，分别 将抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段和抗CTLA-4抗体或抗原结合片段配制成药物组合物，并以适合于所选给药途径的形式向患者给药，例如通过肠胃外、静脉内(iv)、肌肉内、局部或皮下途径。In some embodiments, the anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment are separate administration units and administered in combination. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment can be administered at different times. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered prior to administration of the anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered after administration of the anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, the anti-TIGIT antibody or antigen-binding fragment is administered simultaneously with the anti-CTLA-4 antibody or antigen-binding fragment. In some embodiments, respectively An anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment are formulated into a pharmaceutical composition and administered to the patient in a form suitable for the chosen route of administration, for example, enterally External, intravenous (iv), intramuscular, topical or subcutaneous routes.
在一些实施方案中，抗TIGIT抗体(例如抗体h10D8OF或h10D8OFKF)或抗原结合片段和抗CTLA-4抗体或抗原结合片段同时形成组合给药单元，联合用药。在一些实施方案中，将抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段组合配制成药物组合物，并以适合于所选给药途径的形式向患者给药，例如通过肠胃外、静脉内(iv)、肌肉内、局部或皮下途径。In some embodiments, the anti-TIGIT antibody (eg, antibody h10D8OF or h10D8OFKF) or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment simultaneously form a combined dosage unit for combined administration. In some embodiments, an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment are combined into a pharmaceutical composition and administered to the patient in a form suitable for the chosen route of administration, such as enterally. External, intravenous (iv), intramuscular, topical or subcutaneous routes.
在一些实施方案中，抗TIGIT抗体或抗原结合片段(或制剂)、抗CTLA-4抗体或抗原结合片段(或制剂)可以与其他治疗方法联合使用用于***，例如化疗、放疗和手术治疗等。In some embodiments, anti-TIGIT antibodies or antigen-binding fragments (or formulations), anti-CTLA-4 antibodies or antigen-binding fragments (or formulations) can be used in combination with other treatments to treat tumors, such as chemotherapy, radiation therapy, and surgical treatment. wait.
在一些实施方案中，患者患有肿瘤。在一些实施方案中，肿瘤包括但不限于良性肿瘤、癌症。在一些实施方案中，肿瘤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、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、胰腺癌、***癌、黑色素瘤、头颈癌、肝癌、肾癌、鳞状细胞癌、食管鳞状细胞癌、肺癌、非小细胞肺癌、***、食管癌、子宫内膜癌、卵巢癌、结肠癌、结直肠癌、尿路上皮癌、膀胱癌、脑癌。在一些实施方案中，肿瘤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. In some embodiments, tumors include, but are not limited to, benign tumors, cancer. In some embodiments, tumors include, but are not limited to, hematological cancers, solid tumors. In some embodiments, blood cancers include, but are not limited to, leukemias, lymphomas, and myeloma. In some embodiments, leukemias include acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and myeloid proliferative diseases/neoplasms (MPDS ). In some embodiments, lymphomas include Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, and follicular lymphoma (small cell and large cell). In some embodiments, myeloma includes multiple myeloma (MM), giant cell myeloma, heavy chain myeloma, and light chain or Bence-Jones myeloma. In some embodiments, solid tumors include breast cancer, pancreatic cancer, prostate cancer, melanoma, head and neck cancer, liver cancer, renal cancer, squamous cell carcinoma, esophageal squamous cell carcinoma, lung cancer, non-small cell lung cancer, cervical cancer, Esophageal cancer, endometrial cancer, ovarian cancer, colon cancer, colorectal cancer, urothelial cancer, bladder cancer, brain cancer. In some embodiments, the tumor is a pathologically confirmed locally advanced or metastatic malignant solid tumor for which there is no effective treatment.
在一些实施方案中，抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的组合，其与单独使用抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段治疗相比，所述组合可提高肿瘤抑制作用和/或延长生存期。In some embodiments, the combination of an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment compared to treatment with an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment alone , the combination can improve tumor suppression and/or prolong survival.
附图说明Description of drawings
图1为实施例2的小鼠肿瘤生长曲线。Figure 1 is the mouse tumor growth curve in Example 2.
图2为实施例2的小鼠生存曲线。Figure 2 is the mouse survival curve in Example 2.
图3为实施例3的小鼠肿瘤生长曲线。Figure 3 is the mouse tumor growth curve in Example 3.
图4为实施例3的小鼠生存曲线。Figure 4 is the mouse survival curve in Example 3.
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 stated, 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, e.g. "an antibody" should be understood to mean 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。As used herein, the terms "comprises" or "includes" mean that compositions, methods, and the like include the listed elements, such as components or steps, but do not exclude others.
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 "polypeptide" as well as the plural "polypeptide" and refers to a molecule composed of amino acid monomers linearly linked by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any single 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 two or more amino acid chains, and the term "polypeptide" may Used instead of or interchangeably with any of the above terms. The term "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-natural Amino acid modifications that occur. A polypeptide may be derived from natural biological sources or produced by recombinant techniques, but it does not have to be translated from a specified nucleic acid sequence and 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 an amino group and a carboxyl group, such as an alpha-amino acid, which may 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 the "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 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) with similar chemical properties (eg, charge or hydrophobicity). Generally speaking, conservative amino acid substitutions are unlikely to materially alter the functional properties of the protein. Examples of 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. amino acid; 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 of "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 between any two of these values (inclusive of the endpoints) or any one thereof What value. The number of amino acids of "conservative amino acid substitutions of heavy or light chain" may 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 that is said to "encode" a polypeptide that, in its native state or when manipulated by methods well known to those skilled in the art, is transcribed and/or Or translation can produce 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、抗原结合片段(例如Fab、Fab'、F(ab')2、scFv)。The 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, and antigen-binding fragments. (eg Fab, Fab', F(ab') 2 , 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 a polypeptide or polynucleotide and means a form of the polypeptide or polynucleotide that does not occur in nature, and non-limiting examples may be combined to produce polynucleotides that do not normally exist or Peptides.
“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 positions within each sequence that can be aligned. When a position in the compared sequences is occupied by the same base or amino acid, the molecules are homologous at that position. The degree of homology between sequences is a function of the number of matches 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" means about 80% identity, about 81% identity, about 82% identity, about 83% identity, about 85% identity, about 86% identity, about 87% identity, About 88% identical, about 90% identical, about 91% identical, about 92% identical, about 94% identical, about 95% identical, about 98% identical, about 99% identical, or these The range between any two values in a numeric value, including endpoints, or any value within it.
核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 nucleic acid or polynucleotide sequence (or a polypeptide or antibody sequence) is "identical" or "sequence identical" to another sequence by a certain percentage (eg, 90%, 95%, 98% or 99%). When sequences are aligned, this percentage of bases (or amino acids) in the two sequences being compared are identical. The alignment percent identity or sequence identity can be determined using visual inspection or software programs known in the art, such as those described in Ausubel et al. eds. (2007), Current Protocols in Molecular Biology. It is preferred to use the default parameters for comparison. One of the comparison programs is BLAST using default parameters, such as BLASTN and BLASTP, 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 percentage identity specified above and encode a polypeptide with 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 polypeptides or polypeptide complexes that specifically recognize and bind to antigens. Antibodies can be complete antibodies, 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 of binding to an antigen. anti- Body and antigen-binding fragments include, but are not limited to, complementarity determining regions (CDRs) of heavy or light chains or their ligand-binding portions, heavy chain variable regions (VH), light chain variable regions (VL), heavy chain constant regions (CH), light chain constant region (CL), framework region (FR) or any part thereof, or at least part of a binding protein. The CDR region includes the CDR region of the light chain (L CDR1-3) and the CDR region 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 "antibodies" includes a wide variety of biochemically distinguishable polypeptides. Those skilled in the art will understand that classes of heavy chains include gamma, mu, alpha, delta or epsilon (gamma, mu, alpha, delta, epsilon), of which there are also subclasses (eg γ1-γ4). The nature of this chain determines the "class" of the antibody: IgG, IgM, IgA, IgG, or IgE. Immunoglobulin subclasses (isotypes) such as IgG1, IgG2, IgG3, IgG4, IgG5, etc. are well characterized and the functional specificities conferred are known. All immunoglobulin species are within the scope of the invention. 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Vλ。Light chains can be classified as kappa (κ) or lambda (λ). Each heavy chain can be combined with a kappa or lambda light chain. Generally speaking, 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 joined by covalent disulfide bonds or Non-covalent bonding. In the heavy chain, the amino acid sequence extends from the N-terminus at the fork 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 .
轻链可变区(VL)和重链可变区(VH)决定了抗原识别和特异性。轻链恒定区(CL)和重链恒定区(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；例如IgG1的CH3和CL结构域实际上分别包含重链和轻链的羧基端。The light chain variable region (VL) and heavy chain variable region (VH) determine antigen recognition and specificity. The light chain constant region (CL) and heavy chain constant region (CH) confer important biological properties, such as secretion, transplacental movement, Fc receptor binding, complement binding, etc. By convention, the numbering of constant regions increases as they become farther away from the antibody's antigen-binding site, or amino terminus. The N-terminal part is the variable region and the C-terminal part is the constant region; for example, the CH3 and CL domains of IgG1 actually contain the carboxyl termini 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 a term is used and/or accepted in the art and has two or more definitions, the definition of the term used herein includes all such meanings unless the contrary is expressly stated. A specific example is the use of the term "complementarity determining region" ("CDR") to describe the non-contiguous antigen binding sites found within the variable regions of heavy and light chain polypeptides. This specific area 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., J. Mol. Biol. 196:901-917 (1987). It is incorporated herein 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 defined according to Kabat and Chothia include overlapping 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 numbering comprising 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 CDR contains 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***。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”***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 Proteins of Immunological Interest”(1983)提出的编号***。抗体还可以用EU或Chothia编号***。 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 without relying on experimental data other than the sequence itself. "Kabat numbering" refers to the numbering system proposed by Kabat et al., USDept. of Health and Human Services in "Sequence of Proteins of Immunological Interest" (1983). Antibodies can also use the EU or Chothia numbering system.
“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 preventive or preventative measures designed to prevent, slow down, ameliorate and halt adverse physiological changes or disorders such as the progression of a disease, including but not limited to the following whether detectable or undetectable The results include alleviation of symptoms, reduction in disease severity, stabilization of disease status (i.e. no worsening), delay or slowdown of disease progression, improvement or alleviation of disease status, reduction or disappearance (whether partial or complete), prolongation and Expected survival without treatment, etc. Patients in need of treatment include patients who already have a condition or disorder, are susceptible to a condition or disorder, or are in need of prevention of a condition or disorder from which an antibody or composition disclosed herein can be or is expected to result from administration of an antibody or composition disclosed herein for detection, Patients who benefit from diagnostic procedures and/or treatments.
“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。"Patient" refers to any mammal for whom diagnosis, prognosis or treatment is required, including humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, etc.
“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 conventional error range of the corresponding numerical value that is easily known to those skilled in the relevant technical field. In some embodiments, reference herein to "about" refers to the recited value as well as the range of ±10%, ±5%, or ±1% thereof.
“有效量”是指能引起组织、***、动物、个体或人类的在生物学或医学上反应的活性化合物或药剂的量。"Effective amount" refers to an amount of an active compound or agent that causes a biological or medical response in a tissue, system, animal, individual, or human.
如本文所用，“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。As used herein, "in need" means that a patient has been identified as needing a particular method or treatment. In some embodiments, identification can be by any diagnostic means.
抗体的制备Preparation of antibodies
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尾巴。其他元件包括增强子，Kozak序列及***序列两侧RNA剪接的供体和受***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。常使用的宿主细胞包括HEK293细胞，Cos1细胞，Cos7细胞，CV1细胞，鼠L细胞和CHO细胞等。DNA encoding the antibody can be designed and synthesized according to the antibody amino acid sequence described herein according to conventional methods, placed into an expression vector, and then transfected into host cells, and the transfected host cells are cultured in culture medium to produce monoclonal antibodies. In some embodiments, an expression antibody 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 through the early and late promoters of SV40, the long terminal repeat sequences from retroviruses such as RSV, HTLV1, HIVI, and the early promoter of cytomegalovirus, and other cellular promoters such as muscle can also be used. Kinesin 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 host cells include HEK293 cells, Cos1 cells, Cos7 cells, CV1 cells, mouse L cells and CHO cells.
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 incorporated by reference in their entirety for all purposes.
具体实施方式Detailed ways
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 will be further described below through specific examples, which do not limit the scope of protection of the present invention. Some non-essential modifications and adjustments made by others based on 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。Materials, reagents, etc. used in the following examples can all be obtained from commercial sources unless otherwise specified.
实施例1：抗体的制备Example 1: Preparation of antibodies
将抗体重链和轻链DNA序列克隆到表达载体上，然后转入宿主细胞，培养并纯化获得抗体。The antibody heavy chain and light chain DNA sequences are cloned into expression vectors, then transferred into host cells, cultured and purified to obtain antibodies.
抗体h10D8OF、抗体h10D8OFKF、抗体1、抗体Anti-mCTLA4、抗体Anti-mTIGIT和对照抗体IgG1的氨基酸序列见表1-5，其中，抗体h10D8OF和h10D8OFKF的氨基酸序列是相同的。其中，表达抗体h10D8OFKF和抗体1的宿主细胞为α-(1,6)-岩藻糖基转移酶基因(Fut-8)敲除的CHO-K1细胞，所得的抗体h10D8OFKF岩藻糖基化水平约为0％，抗体1岩藻糖基化水平约为0％，高甘露糖糖型总量<2％，唾液酸化糖型总量<1％。表达抗体h10D8OF的宿主细胞为CHO-K1细胞。表达抗体Anti-mCTLA4、抗体Anti-mTIGIT和对照抗体IgG1的宿主细胞为HEK293F细胞。The amino acid sequences of antibody h10D8OF, antibody h10D8OFKF, antibody 1, antibody Anti-mCTLA4, antibody Anti-mTIGIT and control antibody IgG1 are shown in Table 1-5. Among them, the amino acid sequences of antibodies h10D8OF and h10D8OFKF are the same. Among them, the host cells expressing antibody h10D8OFKF and antibody 1 are CHO-K1 cells with α-(1,6)-fucosyltransferase gene (Fut-8) knockout, and the resulting fucosylation level of antibody h10D8OFKF Approximately 0%, Antibody 1 fucosylation level is approximately 0%, total high mannose glycoforms <2%, and total sialylated glycoforms <1%. The host cells expressing antibody h10D8OF are CHO-K1 cells. The host cells expressing antibody Anti-mCTLA4, antibody Anti-mTIGIT and control antibody IgG1 are HEK293F cells.
表1抗体h10D8OF和h10D8OFKF的氨基酸序列
Table 1 Amino acid sequences of antibodies h10D8OF and h10D8OFKF
表2抗体1的氨基酸序列
Table 2 Amino acid sequence of antibody 1
表3抗体Anti-mCTLA4的氨基酸序列
Table 3 Amino acid sequence of antibody Anti-mCTLA4
表4抗体Anti-mTIGIT的氨基酸序列
Table 4 Amino acid sequence of antibody Anti-mTIGIT
表5对照抗体IgG1的氨基酸序列
Table 5 Amino acid sequence of control antibody IgG1
实施例2：Anti-mTIGIT与Anti-mCTLA4联合给药药效试验Example 2: Efficacy test of combined administration of Anti-mTIGIT and Anti-mCTLA4
本试验评价受试物对Balb/C小鼠皮下荷瘤CT26细胞的治疗效果。在Balb/C小鼠皮下接种CT26结肠癌细胞(接种数量1×106/只)，当小鼠平均肿瘤体积达到大约221mm3时进行分组并给药，分组当天记作第0天，即开始第一次给药，给药方式(给药频率、周期、途径)及具体分组给药方案见表6。试验过程中记录小鼠体重和肿瘤体积，每周2次，并计算肿瘤抑制率TGI％和p值，结果以均值±标准误差表示。当小鼠肿瘤体积大于3000mm3以上时进行安乐死。This test evaluates the therapeutic effect of the test substance on subcutaneous tumor-bearing CT26 cells in Balb/C mice. Balb/C mice were subcutaneously inoculated with CT26 colon cancer cells (inoculation number: 1 × 10 6 /mouse). When the average tumor volume of the mice reached approximately 221 mm 3 , they were divided into groups and administered drugs. The day of grouping was recorded as day 0, which is the start. The first administration, administration mode (administration frequency, cycle, route) and specific group administration regimen are shown in Table 6. During the test, the mouse body weight and tumor volume were recorded twice a week, and the tumor inhibition rate TGI% and p value were calculated. The results were expressed as mean ± standard error. Mice were euthanized when their tumor volume was greater than 3000 mm 3 .
表6给药方式及分组给药方案

Table 6 Dosing methods and grouped dosing schedules

注：BIW：每周2次；i.p.：腹腔注射Note: BIW: twice a week; i.p.: intraperitoneal injection
小鼠肿瘤生长曲线如图1所示，小鼠生存曲线如图2所示。给药后单药组(组2和组3)及联合给药组(组4)都有一定程度的抑制肿瘤的增长，在给药后14天时各组未显示出显著性差异。但随着治疗时间的延长，肿瘤体积增长达到安乐死标准的小鼠都被安乐死，由小鼠的生存曲线可以看出联合给药组(组4)的小鼠生存期明显高于阴性对照组(组1)和单药组(组2和组3)。The mouse tumor growth curve is shown in Figure 1, and the mouse survival curve is shown in Figure 2. After administration, both the single-drug group (Group 2 and Group 3) and the combined administration group (Group 4) inhibited tumor growth to a certain extent, and no significant difference was shown between the groups 14 days after administration. However, with the prolongation of treatment time, mice whose tumor volume reached the euthanasia standard were all euthanized. From the survival curve of the mice, it can be seen that the survival time of the mice in the combined administration group (group 4) was significantly longer than that in the negative control group ( Group 1) and single drug group (Group 2 and Group 3).
实施例3：抗体h10D8OFKF与抗体1联合给药药效试验Example 3: Efficacy test of combined administration of antibody h10D8OFKF and antibody 1
本试验评价受试物在CT26结肠癌细胞皮下移植BALB/c/hTIGIT/hCTLA4小鼠(集萃药康，品系号：T051156)中的抗肿瘤作用，探索抗体h10D8OFKF和抗体1联合给药作用。This test evaluates the anti-tumor effect of the test substance in BALB/c/hTIGIT/hCTLA4 mice (Jicui Yaokang, strain number: T051156) subcutaneously transplanted with CT26 colon cancer cells, and explores the effect of combined administration of antibody h10D8OFKF and antibody 1.
在小鼠皮下接种CT26细胞，接种数量为1×106/只，当小鼠平均肿瘤体积为88.33mm3时进行分组并给药，分组当天记作第0天，即开始第一次给药，给药方式(给药频率、周期、途径)及具体分组给药方案见表7所示。试验过程中记录小鼠体重和肿瘤体积，每周2次，并计算肿瘤抑制率TGI％和p值，结果以均值±标准误差表示。当小鼠肿瘤体积大于3000mm3以上时进行安乐死。Mice were subcutaneously inoculated with CT26 cells at an inoculation number of 1×10 6 /mouse. When the average tumor volume of the mice was 88.33 mm 3 , they were divided into groups and administered drugs. The day of grouping was recorded as day 0, that is, the first administration was started. , the administration method (administration frequency, cycle, route) and the specific group administration scheme are shown in Table 7. During the test, the mouse body weight and tumor volume were recorded twice a week, and the tumor inhibition rate TGI% and p value were calculated. The results were expressed as mean ± standard error. Mice were euthanized when their tumor volume was greater than 3000 mm 3 .
表7给药方式及分组给药方案
Table 7 Dosing methods and grouped dosing schedules
注：BIW：每周2次；i.p.：腹腔注射Note: BIW: twice a week; i.p.: intraperitoneal injection
小鼠肿瘤生长曲线如图3所示，小鼠生存曲线如图4所示。研究结果显示，与对照组对照抗体IgG1相比，受试物抗体1、抗体h10D8OFKF、抗体1与抗体h10D8OFKF联合用药均表现出显著肿瘤抑制作用(见图3)，并可显著延长小鼠生存期(见图4)。与抗体h10D8OFKF单独给药组(组3)相比，抗体1与抗体h10D8OFKF联合给药组(组4)表现出显著肿瘤抑制作用(*，p<0.05)，并可显著延长小鼠生存期(**，p<0.01)。 The mouse tumor growth curve is shown in Figure 3, and the mouse survival curve is shown in Figure 4. The results of the study showed that compared with the control antibody IgG1, the test antibody 1, antibody h10D8OFKF, and the combination of antibody 1 and antibody h10D8OFKF all showed significant tumor inhibitory effects (see Figure 3) and could significantly extend the survival time of mice. (See Figure 4). Compared with the group administered with antibody h10D8OFKF alone (group 3), the group administered antibody 1 in combination with antibody h10D8OFKF (group 4) showed significant tumor inhibition (*, p<0.05) and could significantly prolong the survival of mice ( **, p<0.01).

Claims (34)

  1. 一种联合用药物，包括抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段；所述抗TIGIT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。A combination drug, including an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment; the anti-TIGIT antibody or antigen-binding fragment includes HCDR1 shown in SEQ ID NO:1, SEQ ID NO:2 HCDR2 shown, 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所述的联合用药物，所述抗TIGIT抗体或抗原结合片段包含重链可变区和轻链可变区，其中：The combination drug of claim 1, wherein the anti-TIGIT antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein:
    所述重链可变区包含SEQ ID NO:7所示的氨基酸序列，或与SEQ ID NO:7所示序列相比具有至少80％同一性的氨基酸序列，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 heavy chain variable region includes the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:7, or an amino acid sequence shown in SEQ ID NO:7 Sequence comparison to an amino acid sequence with one or more conservative amino acid substitutions; and/or
    所述轻链可变区包含SEQ ID NO:8所示的氨基酸序列，或与SEQ ID NO:8所示序列相比具有至少80％同一性的氨基酸序列，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 includes the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:8, or an amino acid sequence shown in SEQ ID NO:8 Sequences are compared to amino acid sequences with one or more conservative amino acid substitutions.
  3. 如权利要求1或2所述的联合用药物，所述抗TIGIT抗体的重链包含SEQ ID NO:9所示的氨基酸序列，或与SEQ ID NO:9所示序列相比具有至少80％同一性的氨基酸序列，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 combination drug according to claim 1 or 2, the heavy chain of the anti-TIGIT antibody comprises the amino acid sequence shown in SEQ ID NO:9, or has at least 80% identity with the sequence shown in SEQ ID NO:9. A unique amino acid 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TIGIT抗体的轻链包含SEQ ID NO:10所示的氨基酸序列，或与SEQ ID NO:10所示序列相比具有至少80％同一性的氨基酸序列，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-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 10, or an amino acid sequence having at least 80% identity with the sequence set forth in SEQ ID NO: 10, or an amino acid sequence set forth in SEQ ID NO: 10 The sequences shown are compared to amino acid sequences with one or more conservative amino acid substitutions.
  4. 如权利要求1-3任一项所述的联合用药物，所述抗CTLA-4抗体或抗原结合片段包含SEQ ID NO:11所示的HCDR1、SEQ ID NO:12所示的HCDR2、SEQ ID NO:13所示的HCDR3、SEQ ID NO:14所示的LCDR1、SEQ ID NO:15所示的LCDR2和SEQ ID NO:16所示的LCDR3。The combination drug according to any one of claims 1-3, the anti-CTLA-4 antibody or antigen-binding fragment includes HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, SEQ ID HCDR3 shown in NO:13, LCDR1 shown in SEQ ID NO:14, LCDR2 shown in SEQ ID NO:15 and LCDR3 shown in SEQ ID NO:16.
  5. 如权利要求1-4任一项所述的联合用药物，所述抗CTLA-4抗体或抗原结合片段包含重链可变区和轻链可变区，其中：The combination drug according to any one of claims 1 to 4, wherein the anti-CTLA-4 antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein:
    所述重链可变区包含SEQ ID NO:17所示的氨基酸序列，或与SEQ ID NO:17所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:17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 heavy chain variable region includes the amino acid sequence shown in SEQ ID NO:17, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:17, or an amino acid sequence shown in SEQ ID NO:17 Sequence comparison to an amino acid sequence with one or more conservative amino acid substitutions; and/or
    所述轻链可变区包含SEQ ID NO:18所示的氨基酸序列，或与SEQ ID NO:18所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:18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 comprises the amino acid sequence set forth in SEQ ID NO: 18, or an amino acid sequence having at least 80% identity with the sequence set forth in SEQ ID NO: 18, or an amino acid sequence set forth in SEQ ID NO: 18 The sequence has An amino acid sequence with one or more conservative amino acid substitutions.
  6. 如权利要求1-5任一项所述的联合用药物，所述抗CTLA-4抗体的重链包含SEQ ID NO:19所示的氨基酸序列，或与SEQ ID NO:19所示序列相比具有至少80％同一性的氨基酸序列，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The combination drug according to any one of claims 1-5, the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO: 19, or is compared with the sequence shown in SEQ ID NO: 19 An amino acid sequence with at least 80% identity, 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
    所述抗CTLA-4抗体的轻链包含SEQ ID NO:20所示的氨基酸序列，或与SEQ ID NO:20所示序列相比具有至少80％同一性的氨基酸序列，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。The light chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO: 20, or an amino acid sequence having at least 80% identity with the sequence shown in SEQ ID NO: 20, or with SEQ ID NO: The sequence shown in 20 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
  7. 如权利要求1-6任一项所述的联合用药物，所述抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为1-50:1；或者为5-25:1；或者为约5:1；或者为约25:1。The combined drug according to any one of claims 1 to 6, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-50:1; or 5-25 :1; or about 5:1; or about 25:1.
  8. 一种***的方法，其包括：向有需要的患者给药有效量的抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段；所述抗TIGIT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。A method of treating tumors, comprising: administering an effective amount of an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment to a patient in need; the anti-TIGIT antibody or antigen-binding fragment comprises SEQ ID 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 SEQ ID LCDR3 shown in NO:6.
  9. 如权利要求8所述的方法，所述抗TIGIT抗体或抗原结合片段包含重链可变区和轻链可变区，其中：The method of claim 8, wherein the anti-TIGIT antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein:
    所述重链可变区包含SEQ ID NO:7所示的氨基酸序列，或与SEQ ID NO:7所示序列相比具有至少80％同一性的氨基酸序列，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 heavy chain variable region includes the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:7, or an amino acid sequence shown in SEQ ID NO:7 Sequence comparison to an amino acid sequence with one or more conservative amino acid substitutions; and/or
    所述轻链可变区包含SEQ ID NO:8所示的氨基酸序列，或与SEQ ID NO:8所示序列相比具有至少80％同一性的氨基酸序列，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 includes the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:8, or an amino acid sequence shown in SEQ ID NO:8 Sequences are compared to amino acid sequences with one or more conservative amino acid substitutions.
  10. 如权利要求8或9所述的方法，所述抗TIGIT抗体的重链包含SEQ ID NO:9所示的氨基酸序列，或与SEQ ID NO:9所示序列相比具有至少80％同一性的氨基酸序列，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 8 or 9, wherein the heavy chain of the anti-TIGIT antibody comprises the amino acid sequence shown in SEQ ID NO:9, or has at least 80% identity with the sequence shown in SEQ ID NO:9 Amino acid sequence, 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TIGIT抗体的轻链包含SEQ ID NO:10所示的氨基酸序列，或与SEQ ID NO:10所示序列相比具有至少80％同一性的氨基酸序列，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-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 10, or an amino acid sequence having at least 80% identity with the sequence set forth in SEQ ID NO: 10, or an amino acid sequence set forth in SEQ ID NO: 10. The sequences shown are compared to amino acid sequences with one or more conservative amino acid substitutions.
  11. 如权利要求8-10任一项所述的方法，所述抗CTLA-4抗体或抗原结合片段包含SEQ ID NO:11所示的HCDR1、SEQ ID NO:12所示的HCDR2、SEQ ID NO:13所示的HCDR3、SEQ ID NO:14所示的LCDR1、SEQ ID NO:15所示的LCDR2和SEQ ID NO:16所示的LCDR3。The method according to any one of claims 8-10, the anti-CTLA-4 antibody or antigen-binding fragment comprises HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, SEQ ID NO: HCDR3 shown in SEQ ID NO:13, LCDR1 shown in SEQ ID NO:14, LCDR2 shown in SEQ ID NO:15 and LCDR3 shown in SEQ ID NO:16.
  12. 如权利要求8-11任一项所述的方法，所述抗CTLA-4抗体或抗原结合片段包含重链可变区和轻链可变区，其中：The method of any one of claims 8-11, wherein the anti-CTLA-4 antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein:
    所述重链可变区包含SEQ ID NO:17所示的氨基酸序列，或与SEQ ID NO:17所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:17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 heavy chain variable region includes the amino acid sequence shown in SEQ ID NO:17, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:17, or an amino acid sequence shown in SEQ ID NO:17 Sequence comparison to an amino acid sequence with one or more conservative amino acid substitutions; and/or
    所述轻链可变区包含SEQ ID NO:18所示的氨基酸序列，或与SEQ ID NO:18所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:18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 includes the amino acid sequence shown in SEQ ID NO: 18, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO: 18, or an amino acid sequence shown in SEQ ID NO: 18 Sequences are compared to amino acid sequences with one or more conservative amino acid substitutions.
  13. 如权利要求8-12任一项所述的方法，所述抗CTLA-4抗体的重链包含SEQ ID NO:19所示的氨基酸序列，或与SEQ ID NO:19所示序列相比具有至少80％同一性的氨基酸序列，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The method according to any one of claims 8-12, the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO:19, or has at least An amino acid sequence that is 80% identical, or has one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:19; and/or
    所述抗CTLA-4抗体的轻链包含SEQ ID NO:20所示的氨基酸序列，或与SEQ ID NO:20所示序列相比具有至少80％同一性的氨基酸序列，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。The light chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO: 20, or an amino acid sequence having at least 80% identity with the sequence shown in SEQ ID NO: 20, or with SEQ ID NO: The sequence shown in 20 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
  14. 如权利要求8-13任一项所述的方法，所述抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为1-50:1；或者为5-25:1；或者为约5:1；或者为约25:1。The method according to any one of claims 8-13, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-50:1; or 5-25:1 ; Or about 5:1; Or about 25:1.
  15. 如权利要求8-14任一项所述的方法，所述抗TIGIT抗体或抗原结合片段施用的剂量为0.01mg/kg至20mg/kg或0.05mg至1200mg，所述抗CTLA-4抗体或抗原结合片段施用的剂量为0.01mg/kg至10mg/kg或0.05mg至600mg。The method of any one of claims 8-14, wherein the anti-TIGIT antibody or antigen-binding fragment is administered at a dose of 0.01 mg/kg to 20 mg/kg or 0.05 mg to 1200 mg, and the anti-CTLA-4 antibody or antigen The dosage for administration of the binding fragment is 0.01 mg/kg to 10 mg/kg or 0.05 mg to 600 mg.
  16. 如权利要求8-15任一项所述的方法，所述抗TIGIT抗体或抗原结合片段和/或抗CTLA-4抗体或抗原结合片段的给药频率为每天给药一次至每7周给药一次；或者，给药频率为每天给药一次、每周给药三次、每周给药两次、每周给药一次、每2周给药一次、每3周给药一次、每4周给药一次、每5周给药一次、每6周给药一次或每7周给药一次。The method according to any one of claims 8-15, the administration frequency of the anti-TIGIT antibody or antigen-binding fragment and/or anti-CTLA-4 antibody or antigen-binding fragment ranges from once a day to every 7 weeks. once; alternatively, the dosing frequency is once daily, three times per week, twice per week, once per week, once every 2 weeks, once every 3 weeks, or every 4 weeks. dose once, every 5 weeks, every 6 weeks, or every 7 weeks.
  17. 如权利要求8-16任一项所述的方法，所述肿瘤为良性肿瘤或癌症；或者，所 述肿瘤为血液癌症或实体瘤；或者，所述肿瘤选自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、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、胰腺癌、***癌、黑色素瘤、头颈癌、肝癌、肾癌、鳞状细胞癌、食管鳞状细胞癌、肺癌、非小细胞肺癌、***、食管癌、子宫内膜癌、卵巢癌、多发性骨髓瘤、结肠癌、结直肠癌、尿路上皮癌、膀胱癌、脑癌。The method according to any one of claims 8-16, the tumor is a benign tumor or cancer; or, the The tumor is a blood cancer or a solid tumor; alternatively, the tumor is selected from the group consisting of acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, myeloid proliferative diseases/tumors, and Hodgkin lymphoma , non-Hodgkin lymphoma, Burkitt lymphoma, follicular lymphoma, multiple myeloma, giant cell myeloma, heavy chain myeloma, light chain or Bence-Jones myeloma, breast cancer, pancreatic cancer , prostate cancer, melanoma, head and neck cancer, liver cancer, kidney cancer, squamous cell carcinoma, esophageal squamous cell carcinoma, lung cancer, non-small cell lung cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, multiple myeloma tumors, colon cancer, colorectal cancer, urothelial cancer, bladder cancer, and brain cancer.
  18. ***的药物组合，包括抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段；所述抗TIGIT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。A drug combination for treating tumors, including an anti-TIGIT antibody or antigen-binding fragment and an anti-CTLA-4 antibody or antigen-binding fragment; the anti-TIGIT antibody or antigen-binding fragment includes HCDR1 shown in SEQ ID NO:1, SEQ ID NO:2 HCDR2 shown, 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.
  19. 如权利要求18所述的药物组合，所述抗TIGIT抗体或抗原结合片段包含重链可变区和轻链可变区，其中：The pharmaceutical combination of claim 18, wherein the anti-TIGIT antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein:
    所述重链可变区包含SEQ ID NO:7所示的氨基酸序列，或与SEQ ID NO:7所示序列相比具有至少80％同一性的氨基酸序列，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 heavy chain variable region includes the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:7, or an amino acid sequence shown in SEQ ID NO:7 Sequence comparison to an amino acid sequence with one or more conservative amino acid substitutions; and/or
    所述轻链可变区包含SEQ ID NO:8所示的氨基酸序列，或与SEQ ID NO:8所示序列相比具有至少80％同一性的氨基酸序列，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 includes the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:8, or an amino acid sequence shown in SEQ ID NO:8 Sequences are compared to amino acid sequences with one or more conservative amino acid substitutions.
  20. 如权利要求18或19所述的药物组合，所述抗TIGIT抗体的重链包含SEQ ID NO:9所示的氨基酸序列，或与SEQ ID NO:9所示序列相比具有至少80％同一性的氨基酸序列，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 pharmaceutical combination according to claim 18 or 19, the heavy chain of the anti-TIGIT antibody comprises the amino acid sequence shown in SEQ ID NO:9, or has at least 80% identity with the sequence shown in SEQ ID NO:9 An amino acid sequence, 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TIGIT抗体的轻链包含SEQ ID NO:10所示的氨基酸序列，或与SEQ ID NO:10所示序列相比具有至少80％同一性的氨基酸序列，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-TIGIT antibody comprises the amino acid sequence set forth in SEQ ID NO: 10, or an amino acid sequence having at least 80% identity with the sequence set forth in SEQ ID NO: 10, or an amino acid sequence set forth in SEQ ID NO: 10 The sequences shown are compared to amino acid sequences with one or more conservative amino acid substitutions.
  21. 如权利要求18-20任一项所述的药物组合，所述抗CTLA-4抗体或抗原结合片段包含SEQ ID NO:11所示的HCDR1、SEQ ID NO:12所示的HCDR2、SEQ ID NO:13所示的HCDR3、SEQ ID NO:14所示的LCDR1、SEQ ID NO:15所示的LCDR2和SEQ ID NO:16所示的LCDR3。The pharmaceutical combination according to any one of claims 18-20, wherein the anti-CTLA-4 antibody or antigen-binding fragment includes HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, and SEQ ID NO : HCDR3 shown in SEQ ID NO:13, LCDR1 shown in SEQ ID NO:14, LCDR2 shown in SEQ ID NO:15 and LCDR3 shown in SEQ ID NO:16.
  22. 如权利要求18-21任一项所述的药物组合，所述抗CTLA-4抗体或抗原结合片段包含重链可变区和轻链可变区，其中： The pharmaceutical combination according to any one of claims 18-21, wherein the anti-CTLA-4 antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein:
    所述重链可变区包含SEQ ID NO:17所示的氨基酸序列，或与SEQ ID NO:17所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:17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 heavy chain variable region includes the amino acid sequence shown in SEQ ID NO:17, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO:17, or an amino acid sequence shown in SEQ ID NO:17 Sequence comparison to an amino acid sequence with one or more conservative amino acid substitutions; and/or
    所述轻链可变区包含SEQ ID NO:18所示的氨基酸序列，或与SEQ ID NO:18所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:18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 includes the amino acid sequence shown in SEQ ID NO: 18, or an amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO: 18, or an amino acid sequence shown in SEQ ID NO: 18 Sequences are compared to amino acid sequences with one or more conservative amino acid substitutions.
  23. 如权利要求18-22任一项所述的药物组合，所述抗CTLA-4抗体的重链包含SEQ ID NO:19所示的氨基酸序列，或与SEQ ID NO:19所示序列相比具有至少80％同一性的氨基酸序列，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The pharmaceutical combination according to any one of claims 18-22, the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO: 19, or has an amino acid sequence compared with the sequence shown in SEQ ID NO: 19 An amino acid sequence that is at least 80% identical, or has one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:19; and/or
    所述抗CTLA-4抗体的轻链包含SEQ ID NO:20所示的氨基酸序列，或与SEQ ID NO:20所示序列相比具有至少80％同一性的氨基酸序列，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。The light chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO: 20, or an amino acid sequence having at least 80% identity with the sequence shown in SEQ ID NO: 20, or with SEQ ID NO: The sequence shown in 20 is compared to the amino acid sequence with one or more conservative amino acid substitutions.
  24. 如权利要求18-23任一项所述的药物组合，所述抗TIGIT抗体或抗原结合片段和抗CTLA-4抗体或抗原结合片段的质量比为1-50:1；或者为5-25:1；或者为约5:1；或者为约25:1。The pharmaceutical combination according to any one of claims 18 to 23, the mass ratio of the anti-TIGIT antibody or antigen-binding fragment and the anti-CTLA-4 antibody or antigen-binding fragment is 1-50:1; or 5-25: 1; or about 5:1; or about 25:1.
  25. 权利要求1-7任一项所述的联合用药物在制备用于***的药物中的应用。Use of the combined drug according to any one of claims 1 to 7 in the preparation of drugs for treating tumors.
  26. 权利要求1-7任一项所述的联合用药物或权利要求18-24任一项所述的药物组合在***中的应用。Application of the combined drug according to any one of claims 1 to 7 or the drug combination according to any one of claims 18 to 24 in the treatment of tumors.
  27. 如权利要求25或26所述的应用，所述肿瘤为良性肿瘤或癌症；或者，所述肿瘤为血液癌症或实体瘤；或者，所述肿瘤选自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、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、胰腺癌、***癌、黑色素瘤、头颈癌、肝癌、肾癌、鳞状细胞癌、食管鳞状细胞癌、肺癌、非小细胞肺癌、***、食管癌、子宫内膜癌、卵巢癌、多发性骨髓瘤、结肠癌、结直肠癌、尿路上皮癌、膀胱癌、脑癌。The application according to claim 25 or 26, the tumor is a benign tumor or cancer; or the tumor is a blood cancer or a solid tumor; or the tumor is selected from acute lymphoblastic leukemia, acute myelogenous leukemia, Chronic lymphocytic leukemia, chronic myelogenous leukemia, myeloid proliferative disorders/neoplasms, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, follicular lymphoma, multiple myeloma, giant cell Myeloma, heavy chain myeloma, light chain or Bence-Jones myeloma, breast cancer, pancreatic cancer, prostate cancer, melanoma, head and neck cancer, liver cancer, kidney cancer, squamous cell carcinoma, esophageal squamous cell carcinoma, lung cancer , non-small cell lung cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, multiple myeloma, colon cancer, colorectal cancer, urothelial cancer, bladder cancer, brain cancer.
  28. 一种试剂盒，其包含抗TIGIT抗体和包装插页，该包装插页包含关于与抗CTLA-4抗体组合使用所述抗TIGIT抗体***的说明书。A kit comprising an anti-TIGIT antibody and a package insert containing instructions for using the anti-TIGIT antibody in combination with an anti-CTLA-4 antibody to treat tumors.
  29. 一种试剂盒，其包含抗CTLA-4抗体和包装插页，该包装插页包含关于与抗TIGIT抗体组合使用所述抗CTLA-4抗体***的说明书。 A kit comprising an anti-CTLA-4 antibody and a package insert containing instructions for using the anti-CTLA-4 antibody in combination with an anti-TIGIT antibody to treat tumors.
  30. 一种试剂盒，其包含抗TIGIT抗体和抗CTLA-4抗体的药剂，和包装插页，该包装插页包含关于使用所述抗TIGIT抗体和抗CTLA-4抗体的药剂***的说明书。A kit comprising a medicament of an anti-TIGIT antibody and an anti-CTLA-4 antibody, and a package insert containing instructions for treating tumors using the medicament of an anti-TIGIT antibody and an anti-CTLA-4 antibody.
  31. 一种试剂盒，其包含抗TIGIT抗体的药剂和包装插页，该包装插页包含关于与抗CTLA-4抗体组合使用所述抗TIGIT抗体的药剂***的说明书。A kit comprising a medicament of an anti-TIGIT antibody and a package insert containing instructions for using the medicament of an anti-TIGIT antibody in combination with an anti-CTLA-4 antibody to treat tumors.
  32. 一种试剂盒，其包含抗CTLA-4抗体的药剂和包装插页，该包装插页包含关于与抗TIGIT抗体组合使用所述抗CTLA-4抗体的药剂***的说明书。A kit comprising a medicament of an anti-CTLA-4 antibody and a package insert containing instructions for using the medicament of an anti-CTLA-4 antibody in combination with an anti-TIGIT antibody to treat tumors.
  33. 如权利要求28-32任一项所述的试剂盒，所述抗TIGIT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；优选地，所述抗TIGIT抗体或抗原结合片段包含重链可变区和轻链可变区，其中所述重链可变区包含SEQ ID NO:7所示的氨基酸序列，或与SEQ ID NO:7所示序列相比具有至少80％同一性的氨基酸序列，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/或所述轻链可变区包含SEQ ID NO:8所示的氨基酸序列，或与SEQ ID NO:8所示序列相比具有至少80％同一性的氨基酸序列，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；优选地，所述抗TIGIT抗体的重链包含SEQ ID NO:9所示的氨基酸序列，或与SEQ ID NO:9所示序列相比具有至少80％同一性的氨基酸序列，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/或所述抗TIGIT抗体的轻链包含SEQ ID NO:10所示的氨基酸序列，或与SEQ ID NO:10所示序列相比具有至少80％同一性的氨基酸序列，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 kit according to any one of claims 28-32, the anti-TIGIT antibody or antigen-binding fragment includes HCDR1 shown in SEQ ID NO:1, HCDR2 shown in SEQ ID NO:2, and SEQ ID NO:3 HCDR3 shown, LCDR1 shown in SEQ ID NO:4, LCDR2 shown in SEQ ID NO:5 and LCDR3 shown in SEQ ID NO:6; Preferably, the anti-TIGIT antibody or antigen-binding fragment includes a heavy chain Variable region and light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7, or an amino acid with at least 80% identity compared to the sequence set forth in SEQ ID NO:7 sequence, or an amino acid sequence having one or more conservative amino acid substitutions compared with the sequence shown in SEQ ID NO:7; and/or the light chain variable region includes the amino acid sequence shown in SEQ ID NO:8, or with An amino acid sequence with at least 80% identity compared to the sequence shown in SEQ ID NO: 8, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 8; Preferably, the antibody The heavy chain of the TIGIT antibody contains the amino acid sequence set forth in SEQ ID NO:9, or an amino acid sequence that is at least 80% identical to the sequence set forth in SEQ ID NO:9, or is similar to the sequence set forth in SEQ ID NO:9 Compared with an amino acid sequence having one or more conservative amino acid substitutions; and/or the light chain of the anti-TIGIT antibody comprises the amino acid sequence shown in SEQ ID NO:10, or has at least An amino acid sequence that is 80% identical, or has one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO:10.
  34. 如权利要求28-33任一项所述的试剂盒，所述抗CTLA-4抗体或抗原结合片段包含SEQ ID NO:11所示的HCDR1、SEQ ID NO:12所示的HCDR2、SEQ ID NO:13所示的HCDR3、SEQ ID NO:14所示的LCDR1、SEQ ID NO:15所示的LCDR2和SEQ ID NO:16所示的LCDR3；优选地，所述抗CTLA-4抗体或抗原结合片段包含重链可变区和轻链可变区，其中所述重链可变区包含SEQ ID NO:17所示的氨基酸序列，或与SEQ ID NO:17所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:17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/或所述轻链可变区包含SEQ ID NO:18所示的氨基酸序列，或与SEQ ID NO:18所示序列相比具有至少80％同一性的氨基酸序列，或与SEQ ID NO:18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；优选地，所述抗CTLA-4抗体的重链包含SEQ ID NO:19所示的氨基酸序列，或与SEQ ID NO:19所示序列相比具有至少80％同一性的氨基酸序列， 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CTLA-4抗体的轻链包含SEQ ID NO:20所示的氨基酸序列，或与SEQ ID NO:20所示序列相比具有至少80％同一性的氨基酸序列，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。 The kit according to any one of claims 28 to 33, wherein the anti-CTLA-4 antibody or antigen-binding fragment includes HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, and SEQ ID NO : HCDR3 shown in SEQ ID NO:13, LCDR1 shown in SEQ ID NO:14, LCDR2 shown in SEQ ID NO:15 and LCDR3 shown in SEQ ID NO:16; Preferably, the anti-CTLA-4 antibody or antigen binds The fragment comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 17, or has at least 80% compared to the sequence set forth in SEQ ID NO: 17 An identical amino acid sequence, or an amino acid sequence with one or more conservative amino acid substitutions compared to the sequence set forth in SEQ ID NO: 17; and/or the light chain variable region includes the amino acid set forth in SEQ ID NO: 18 Sequence, or an amino acid sequence having at least 80% identity compared to the sequence shown in SEQ ID NO: 18, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 18; preferably , the heavy chain of the anti-CTLA-4 antibody comprises the amino acid sequence shown in SEQ ID NO: 19, or an amino acid sequence having at least 80% identity compared to the sequence shown in SEQ ID NO: 19, Or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 19; and/or the light chain of the anti-CTLA-4 antibody includes the amino acid sequence shown in SEQ ID NO: 20, or An amino acid sequence that is at least 80% identical to the sequence shown in SEQ ID NO: 20, or an amino acid sequence that has one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 20.
PCT/CN2023/115904 2022-08-31 2023-08-30 Combined use of anti-tigit antibody and anti-ctla-4 antibody in treating tumor WO2024046389A1 (en)

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