Methods of treating lymphoma using anti-TIGIT antibodies

Abstract

Methods of treating diffuse large B-cell lymphoma (DLBCL) or increasing, enhancing, or stimulating an immune response with antibodies and antigen-binding fragments thereof that specifically bind to TIGIT (T cell immune receptors with Ig and ITIM domains) in combination with an anti-PD1 antibody and / or an anti-CD20 antibody are provided.

Description

Technical Field

[0001] The present application relates to a combination of an antibody that specifically binds to TIGIT (a T cell immunoreceptor having Ig and ITIM domains) with an anti-PD1 antibody and / or an anti-CD20 antibody for the treatment of lymphoma, such as diffuse large B-cell lymphoma (DLCBL). Background Art

[0002] TIGIT (T cell immunoglobulin and ITIM domain) is a type I transmembrane protein and a member of the CD28 protein family. It plays an important role in inhibiting T cell and NK cell-mediated functional activities in anti-tumor immunity (Boles KS et al., 2009 Eur J Immunol, 39: 695-703; Stanietsky N et al., 2009 PNAS, 106: 17858-63; Yu X et al., 2009 Nat. Immunol, 10: 48-57).

[0003] The gene and cDNA encoding TIGIT were cloned and characterized in mice and humans. Full-length human TIGIT is a sequence of 244 amino acids in length (SEQ ID NO: 1), of which the first 21 amino acids constitute a signal peptide. The amino acid sequence of mature human TIGIT contains 223 amino acid (aa) residues (NCBI accession number: NM_173799). The extracellular domain (ECD) of mature human TIGIT consists of 120 amino acid residues (SEQ ID NO: 2, corresponding to amino acids 22-141 of SEQ ID NO: 1) with a V-type Ig-like domain (corresponding to amino acids 39-127 of SEQ ID NO: 1), followed by a 21-amino acid transmembrane sequence and an 82-amino acid cytoplasmic domain with an immunoreceptor tyrosine-based inhibitory motif (ITIM) (Yu X et al. 2009 Nat. Immunol [Natural Immunology], 10: 48-57; Stengel KF et al. 2012 PNAS [Proceedings of the National Academy of Sciences of the United States of America] 109: 5399-04). In the ECD, human TIGIT has only 59% and 87% amino acid sequence identity with mice and cynomolgus monkeys, respectively.

[0004] Monoclonal antibodies targeting PD1 or PD-L1 can block this interaction and enhance the immune response against cancer cells. These antibodies have been shown to help treat several types of cancer, including skin melanoma, non-small cell lung cancer (NSCLC), kidney cancer, bladder cancer, head and neck cancer, and Hodgkin lymphoma. Most cancer cells that do not respond to single-agent checkpoint inhibitors escape through innate mechanisms that allow cancer cells to grow and survive. As a result, the disease progresses at a rate consistent with the natural history. However, unlike intrinsic resistance, patients with previous clinical benefit now have late relapses after longer follow-up in clinical trials, indicating the emergence of acquired resistance (Jenkins et al., Br. J. Cancer [British Journal of Cancer] 118, 9-16 2018).

[0005] Anti-CD20 antibodies (e.g., rituximab) can be responsible for the rapid killing of tumor targets via FcγR-mediated ADCC or antibody-dependent cellular phagocytosis (ADCP), a short-term process mediated by natural killer cells and myeloid effector cells of the innate immune system (Taylor and Lindorfer, Curr Opin Immunol. 2008; 20(4): 444-449; Minard-Colin et al., Blood 2008; 112(4): 1205-1213). Studies have also revealed that the therapeutic function and long-term immune memory of anti-CD20 antibodies require active interaction with adaptive immunity. Evidence from lymphoma patients suggests that rituximab can induce a strong anti-tumor vaccine effect by eliciting anti-tumor T cell responses, resulting in a long-lasting response (Cartron et al., Blood 2004; 104(9): 2635-2642). Using a murine lymphoma model expressing human CD20, it was demonstrated that CD4+ and CD8+ T cells are required for long-term memory immune responses mediated by anti-CD20 mAbs (Abes et al., Blood. 2010; 116(6):926-934), and the mechanism of anti-CD20 antibodies was shown to be to engage macrophages for immediate ADCC and dendritic cells for long-term anti-tumor cellular immune responses (DiLillo et al., Cell. 2015; 161(5):1035-1045).

[0006] Diffuse large B-cell lymphoma (DLBCL) is an aggressive, heterogeneous disease with variable clinical outcomes (Susanibar-Adaniya et al., Am J Hematol. 2021; 96(5): 617-629) and is the most common subtype of aggressive non-Hodgkin lymphoma (NHL) in Western countries, accounting for approximately 31% of adult NHL cases. In 2020, there were 544,352 new cases of NHL worldwide and 259,793 deaths from the disease (Sung et al., CA Cancer J Clin. 2021; 71(3): 209-249). The incidence of DLBCL is approximately 6.9 cases per 100,000 person-years (Teras et al., CA Cancer J Clin. 2016 Nov 12; 66(6): 443-459). Based on the research report of the National Cancer Registry of China, in 2015, China is expected to have 88,200 new cases of lymphoma and 52,100 deaths from lymphoma, ranking 12th and 11th among all cancer cases, respectively (Chen et al., CA Cancer J Clin. 2016; 66(2):115-132). Two local retrospective studies on the distribution of NHL in China showed that DLBCL accounted for approximately 29.1% and 40.9% of NHL, respectively (Liu et al., Asian Pac J Cancer Prev. 2011; 12(11):3055-3061; Gross et al., China. Int J Hematol. 2008; 88(2):165-173). According to the current Surveillance, Epidemiology, and End Results (SEER) data, the median age at diagnosis is 66 years (SEER 2020). Therefore, the combination of anti-TIGIT antibodies and anti-PD1 antibodies can avoid the development of tolerance in immune cells and induce effective immune responses in the treatment of cancer or chronic viral infections. Summary of the invention

[0007] The present disclosure relates to methods of treating diffuse large B-cell lymphoma (DLBCL) administering an anti-TIGIT antibody in combination with an anti-PD1 antibody and / or an anti-CD20 antibody.

[0008] A method for treating diffuse large B-cell lymphoma (DLBCL), the method comprising administering to a subject an effective amount of an anti-TIGIT antibody or an antigen-binding fragment thereof in combination with an effective amount of an anti-PD1 antibody or an antigen-binding fragment thereof.

[0009] The method, wherein the method comprises administering to a subject an effective amount of an antibody or an antigen-binding fragment thereof, which antibody or antigen-binding fragment thereof specifically binds to human TIGIT and comprises: a heavy chain variable region, which comprises HCDR (heavy chain complementarity determining region) 1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3; and a light chain variable region, which comprises LCDR (light chain complementarity determining region) 1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6.

[0010] The method, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises: a heavy chain variable region (VH) comprising SEQ ID NO:7 and a light chain variable region (VL) comprising SEQ ID NO:8.

[0011] The method, wherein the anti-PD1 antibody comprises an antibody or an antigen-binding fragment thereof, which specifically binds to human PD1 and comprises: A heavy chain variable region comprising: HCDR1 of SEQ ID NO:12, HCDR2 of SEQ ID NO:13, and HCDR3 of SEQ ID NO:14; and a light chain variable region comprising LCDR1 of SEQ ID NO:15, LCDR2 of SEQ ID NO:16, and LCDR3 of SEQ ID NO:17.

[0012] The method, wherein the anti-PD1 antibody or its antigen-binding fragment specifically binds to human PD1 and comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 18 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 19.

[0013] The method, wherein the anti-PD1 antibody comprises an IgG4 constant domain comprising SEQ ID NO:20.

[0014] The method, wherein the anti-TIGIT antibody is an antibody fragment selected from the group consisting of: Fab, Fab'-SH, Fv, scFv, and (Fab')2 fragment.

[0015] The method, wherein the anti-PD1 antibody is an antibody fragment selected from the group consisting of: Fab, Fab'-SH, Fv, scFv, and (Fab')2 fragment.

[0016] A method for treating diffuse large B-cell lymphoma (DLBCL), the method comprising administering to a subject an effective amount of an anti-TIGIT antibody or an antigen-binding fragment thereof in combination with an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof.

[0017] The method, wherein the method comprises administering to a subject an effective amount of an antibody or an antigen-binding fragment thereof, which antibody or antigen-binding fragment thereof specifically binds to human TIGIT and comprises: a heavy chain variable region, which comprises HCDR (heavy chain complementarity determining region) 1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3; and a light chain variable region, which comprises LCDR (light chain complementarity determining region) 1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6.

[0018] The method, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises: a heavy chain variable region (VH) comprising SEQ ID NO:7 and a light chain variable region (VL) comprising SEQ ID NO:8.

[0019] The method, wherein the anti-CD20 antibody is rituximab (Rituxan), ibritumomab tiuxetan (Zevalin), tositumomab (Bexxar), or obinutuzumab (Gazyva).

[0020] The method, wherein the anti-CD20 antibody is rituximab (Rituxan).

[0021] The method, wherein the DLBCL is relapsed or refractory DLBCL.

[0022] The method further comprises administering chemotherapy.

[0023] The method wherein the chemotherapy is cyclophosphamide, doxorubicin, vincristine, and prednisone.

[0024] The method, wherein the anti-PD1 antibody is administered at 200 mg every three weeks.

[0025] The method, wherein the anti-TIGIT antibody is administered in the range of 50 mg-900 mg.

[0026] The method, wherein the anti-TIGIT antibody is administered at 50 mg every three weeks.

[0027] The method, wherein the anti-TIGIT antibody is administered at 150 mg every three weeks.

[0028] The method, wherein the anti-TIGIT antibody is administered at 450 mg every three weeks.

[0029] The method, wherein the anti-TIGIT antibody is administered at 900 mg every three weeks.

[0030] This approach, in which rituximab is given at 375 mg / m 2 Application.

[0031] A method for treating diffuse large B-cell lymphoma (DLBCL), the method comprising administering to a subject an effective amount of an anti-TIGIT antibody or an antigen-binding fragment thereof in combination with an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof and an effective amount of an anti-PD1 antibody.

[0032] The method, wherein the method comprises administering to a subject an effective amount of an antibody or an antigen-binding fragment thereof, which antibody or antigen-binding fragment thereof specifically binds to human TIGIT and comprises: a heavy chain variable region, which comprises HCDR (heavy chain complementarity determining region) 1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3; and a light chain variable region, which comprises LCDR (light chain complementarity determining region) 1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6.

[0033] The method, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises: a heavy chain variable region (VH) comprising SEQ ID NO:7 and a light chain variable region (VL) comprising SEQ ID NO:8.

[0034] The method, wherein the anti-PD1 antibody comprises an antibody or an antigen-binding fragment thereof, which specifically binds to human PD1 and comprises: a heavy chain variable region, which comprises: HCDR1 of SEQ ID NO: 12, HCDR2 of SEQ ID NO: 13, and HCDR3 of SEQ ID NO: 14; and a light chain variable region, which comprises LCDR1 of SEQ ID NO: 15, LCDR2 of SEQ ID NO: 16, and LCDR3 of SEQ ID NO: 17.

[0035] The method, wherein the anti-PD1 antibody or its antigen-binding fragment specifically binds to human PD1 and comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 18 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 19.

[0036] The method further comprises administering chemotherapy.

[0037] The method wherein the chemotherapy is cyclophosphamide, doxorubicin, vincristine, and prednisone. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 is a graphical representation of the therapeutic agents, their doses, and cohorts. DETAILED DESCRIPTION definition

[0039] Conservative amino acid substitutions of amino acids are well known in the art and are exemplarily shown in the table below. In general, conservative amino acid substitutions mean that the amino acid residue is replaced by another amino acid residue with a similar side chain.

[0040] Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0041] As used herein, including the appended claims, singular forms such as "a," "an," and "the" include their corresponding plural referents unless the context clearly dictates otherwise.

[0042] Unless the context clearly indicates otherwise, the term "or" means and is used interchangeably with the term "and / or".

[0043] Throughout this specification and the following claims, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be understood to implicitly include the stated amino acid sequence, DNA sequence, steps or groups thereof, but not to exclude any other amino acid sequence, DNA sequence, steps. When used herein, the term "comprising" may be replaced with the term "containing" or "including", or sometimes with "having".

[0044] The term "TIGIT" includes various mammalian isoforms, such as human TIGIT, homologs of human TIGIT, and analogs comprising at least one epitope within TIGIT. The amino acid sequence of TIGIT (eg, human TIGIT) and the nucleotide sequence encoding the TIGIT are known in the art (see Genbank AAI01289).

[0045] "PD1" or "programmed cell death protein 1" is a receptor that delivers inhibitory signals upon binding to the ligands CD274 / PD-L1 and CD273 / PD-L2. The amino acid sequence of human PD1 is known in the art and can be found in accession number NP_005009 (Genbank).

[0046] The term "CD-20" or "B lymphocyte surface antigen B1" is a B lymphocyte-specific membrane protein that helps regulate cellular calcium influx. The amino acids of CD20 are shown in Genbank Accession No. NP_068769.

[0047] As used herein, the terms "administration / administering" and "treating / treatment", when applied to an animal, a human, an experimental subject, a cell, a tissue, an organ, or a biological fluid, mean contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of cells encompasses contact of an agent with a cell and contact of an agent with a fluid, wherein the fluid is in contact with a cell. The terms "administration" or "treatment" also include in vitro and ex vivo treatments of, for example, cells by an agent, a diagnostic agent, a binding compound, or another cell. The term "subject" herein refers to any organism, preferably an animal, more preferably a mammal (e.g., rat, mouse, dog, cat, rabbit), and most preferably a human.

[0048] The term "antibody" is used in the broadest sense herein and specifically encompasses antibodies (including full-length monoclonal antibodies) and antibody fragments as long as they recognize an antigen, such as TIGIT. Antibodies are typically monospecific, but can also be described as having different specificities, heterospecificity, or multispecificity. Antibody molecules bind to specific antigenic determinants or epitopes on an antigen through specific binding sites.

[0049] The term "monoclonal antibody" or "mAb" or "Mab" herein means a population of substantially homogeneous antibodies, that is, the antibody molecules contained in the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in small amounts. In contrast, conventional (polyclonal) antibody preparations typically include a variety of different antibodies with different amino acid sequences in their variable domains, particularly their complementary determining regions (CDRs), which are typically specific for different epitopes. The modifier "monoclonal" indicates the characteristics of antibodies obtained from a substantially homogeneous antibody population and should not be construed as requiring antibodies to be produced by any specific method. Monoclonal antibodies (mAbs) can be obtained by methods known to those skilled in the art. See, e.g., Kohler G et al., Nature 1975 256:495-497; U.S. Pat. No. 4,376,110; Ausubel FM et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY 1992; Harlow E et al., ANTIBODIES: A LABORATORY MANUAL, Cold spring Harbor Laboratory 1988; and Colligan JE et al., CURRENT PROTOCOLS IN IMMUNOLOGY 1993. The mAbs disclosed herein can be of any immunoglobulin class, including IgG, IgM, IgD, IgE, IgA, and any subclass thereof. The hybridoma producing the mAb can be cultured in vitro or in vivo. High titer mAbs can be obtained by in vivo production, in which cells from a single hybridoma are injected intraperitoneally into mice, such as primary primed Balb / c mice, to produce ascites fluid containing high concentrations of the desired mAb. MAbs of isotype IgM or IgG can be purified from such ascites fluid, or from culture supernatants, using column chromatography methods well known to those skilled in the art.

[0050] Typically, the basic antibody structural unit comprises a tetramer. Each tetramer includes two pairs of identical polypeptide chains, each pair having a "light chain" (about 25 kDa) and a "heavy chain" (about 50-70 kDa). The amino terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids that is primarily responsible for antigen recognition. The carboxyl terminal portion of the heavy chain can be defined as a constant region that is primarily responsible for effector function. Typically, human light chains are classified as κ and λ light chains. In addition, human heavy chains are typically classified as α, δ, ε, γ or μ, and the isotype of the antibody is defined as IgA, IgD, IgE, IgG and IgM, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of about 12 or more amino acids, and the heavy chain also includes a "D" region of about 10 or more amino acids.

[0051] The variable regions of each light chain / heavy chain (VL / VH) pair form an antibody binding site. Therefore, in general, a complete antibody has two binding sites. Except for bifunctional or bispecific antibodies, in general, the two binding sites are identical.

[0052] Typically, the variable domains of the heavy and light chains contain three hypervariable regions, also referred to as "complementarity determining regions (CDRs)", which are located between relatively conserved framework regions (FRs). The CDRs are usually aligned by the framework regions to enable binding to specific epitopes. In general, from N-terminus to C-terminus, both the light and heavy chain variable domains contain FR-1 (or FR1), CDR-1 (or CDR1), FR-2 (FR2), CDR-2 (CDR2), FR-3 (or FR3), CDR-3 (CDR3) and FR-4 (or FR4) in order. Generally, the amino acid assignments for each domain conform to the definition of an immunologically interesting protein sequence, Kabat et al., National Institutes of Health, Bethesda, Maryland; 5th ed.; NIH Publication No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32: 1-75; Kabat et al., (1977) J. Biol. Chem. 252: 6609-6616; Chothia et al., (1987) J Mol. Biol. 196: 901-917 or Chothia et al., (1989) Nature 342: 878-883.

[0053] The term "hypervariable region" means the amino acid residues in an antibody that are responsible for antigen binding. The hypervariable region comprises amino acid residues from the "CDRs" (i.e., VL-CDR1, VL-CDR2, and VL-CDR3 in the light chain variable domain and VH-CDR1, VH-CDR2, and VH-CDR3 in the heavy chain variable domain). See, Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Maryland (defining the CDR regions of antibodies by sequence); see also Chothia and Lesk (1987) J. Mol. Biol. 196:901-917 (defining the CDR regions of antibodies by structure). The term "framework" or "FR" residues is intended to refer to those variable domain residues other than the hypervariable region residues herein defined as CDR residues.

[0054] Unless otherwise indicated, "antibody fragment" or "antigen-binding fragment" means an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to specifically bind to an antigen bound by a full-length antibody, such as a fragment that retains one or more CDR regions. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules, e.g., single-chain Fv (ScFv); nanobodies, and multispecific antibodies formed from antibody fragments.

[0055] Antibodies that specifically bind to a specific target protein are also described as specifically binding to a specific target protein. This means that compared with other proteins, the antibody shows preferential binding to the target protein, but this specificity does not require absolute binding specificity. If the binding of the antibody determines the presence of the target protein in the sample, for example, no undesirable results, such as false positives, are produced, the antibody is considered to be "specific" for its intended target. The antibodies or their binding fragments that can be used in the present invention will bind to the target protein with an affinity that is at least twice higher than that of the non-target protein, preferably at least 10 times higher, more preferably at least 20 times higher, and most preferably at least 100 times higher. The antibodies herein are referred to as specifically binding to a polypeptide comprising a given amino acid sequence (e.g., the amino acid sequence of a mature human TIGIT molecule), if it binds to a polypeptide comprising the sequence but does not bind to a protein lacking the sequence.

[0056] The expressions "pH-dependent binding", "pH-dependent target binding" and "pH-dependent antigen binding" are interchangeable in the present disclosure, indicating that the antibodies of the present application bind to their targets / antigens, i.e., human TIGIT, in a pH-dependent manner. Specifically, the antibodies of the present application exhibit higher binding affinity and / or binding signals to their antigens at weakly acidic pH values ​​(e.g., pH 6.0, which is typically present in the tumor microenvironment) compared to the binding affinity and / or binding signals at physiological pH (e.g., pH 7.4). Methods for determining the binding affinity and / or binding signal intensity of the antibodies of the present application are well known in the art, including but not limited to surface plasmon resonance (Biacore) or similar techniques. More specifically, as measured by surface plasmon resonance (Biacore) or similar techniques, the antibodies of the present application have a K value greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more at pH 7.4 / pH 6.0. D Ratio. Alternatively or additionally, the Rmax (RU) value of the antibody of the present application at pH 6.0 is at least 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 20 times, 30 times, 40 times, 50 times higher than the Rmax at pH 7.4 measured by surface plasmon resonance (Biacore) or similar techniques. The binding affinity of the antibody can be measured at 25°C or 37°C. It has been found that the tumor microenvironment shows a relatively more acidic pH than physiological conditions or normal tissues. Therefore, the antibody of the present application with the above-mentioned pH-dependent binding is advantageous as an anti-TIGIT therapeutic agent for selectively targeting TIGIT-positive lymphocytes in the tumor microenvironment, and has lower toxicity associated with peripheral activation of lymphocytes.

[0057] The term "human antibody" as used herein means an antibody that comprises only human immunoglobulin protein sequences. If produced in mice, mouse cells, or hybridomas derived from mouse cells, human antibodies may contain murine carbohydrate chains. Similarly, "mouse antibody" or "rat antibody" means an antibody that comprises only mouse or rat immunoglobulin protein sequences, respectively.

[0058] The term "humanized antibody" means an antibody form that contains sequences from non-human (e.g., mouse) antibodies as well as human antibodies. Such antibodies contain minimal sequences derived from non-human immunoglobulins. Usually, a humanized antibody will contain essentially all of at least one, and typically two, variable domains, all or essentially all of its hypervariable loops corresponding to those of non-human immunoglobulins, and all or essentially all of the FRs being those of human immunoglobulin sequences. The humanized antibody will also optionally contain at least a portion of an immunoglobulin constant region (Fc), typically at least a portion of a human immunoglobulin. When it is necessary to distinguish a humanized antibody from a parent rodent antibody, the prefix " hum "," hu ”, “Hu” or “h” is added to the antibody clone name. A humanized form of a rodent antibody will generally contain the same CDR sequences of the parent rodent antibody, but may include certain amino acid substitutions to increase affinity, increase the stability of the humanized antibody, or for other reasons.

[0059] The antibodies of the present application have potential therapeutic uses in treating cancer. The term "cancer" or "tumor" herein refers to or describes a physiological condition in which a mammal is typically characterized by dysregulated cell growth. Examples of cancer include, but are not limited to, leukemia, lymphoma, diffuse large B-cell lymphoma (DLBCL), and relapsed or refractory DLBCL.

[0060] In addition, the antibodies of the present application have potential therapeutic uses in controlling viral infections and other human diseases that are mechanistically related to immune tolerance or "exhaustion." In the context of the present application, the term "exhaustion" refers to a process that causes the exhaustion of the ability of immune cells to respond during cancer or chronic viral infection.

[0061] As used herein, the term "therapeutically effective amount" refers to an amount of antibody sufficient to affect the treatment of a disease, disorder or symptom when applied to a subject to treat at least one clinical symptom of a disease or disorder or disorder. A "therapeutically effective amount" can vary with the severity of the symptoms of the antibody, disease, disorder, and / or disease or disorder, the age of the subject to be treated, and / or the weight of the subject to be treated. The appropriate amount in any given case is apparent to those skilled in the art, or can be determined by routine experiments. In the case of combination therapy, a "therapeutically effective amount" refers to the total amount of the activating agent contained in the combination for the effective treatment of a disease, disorder or condition.

[0062] As used herein, a "subject" is a mammal, eg, a rodent or a primate, preferably a higher primate, such as a human (eg, a patient having or at risk of having a disorder described herein). Anti-TIGIT antibodies

[0063] The present disclosure provides antibodies and antigen-binding fragments that specifically bind to human TIGIT. In addition, the present disclosure provides antibodies with desired pharmacokinetic characteristics and other desired properties, and can therefore be used to treat lymphomas (e.g., DLBCL). The present disclosure further provides pharmaceutical compositions comprising antibodies and methods for preparing and using such pharmaceutical compositions to prevent and treat DLBCL and related disorders.

[0064] The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to TIGIT. Anti-TIGIT antibodies are also provided herein and include, for example, a heavy chain variable region (VH) comprising a complementary determining region (CDR): HCDR1 as shown in SEQ ID NO: 1, HCDR2 as shown in SEQ ID NO: 2, and HCDR3 as shown in SEQ ID NO: 3; and a light chain variable region (VL), comprising: LCDR1 as shown in SEQ ID NO: 4, LCDR2 as shown in SEQ ID NO: 5, and LCDR3 as shown in SEQ ID NO: 6. The antibodies or antigen-binding fragments of the present disclosure include, but are not limited to, antibodies or antigen-binding fragments thereof as described in Table 1 below, which are designated as "BGB-A1217" or "Ociperlimab". Table 1 Anti-PD1 antibodies

[0065] PD1 antibodies are also provided herein and include, for example, a heavy chain variable region (VH) comprising a complementarity determining region (CDR): HCDR1 as shown in SEQ ID NO: 12, HCDR2 as shown in SEQ ID NO: 13, and HCDR3 as shown in SEQ ID NO: 14; and a light chain variable region (VL) comprising: LCDR1 as shown in SEQ ID NO: 15, LCDR2 as shown in SEQ ID NO: 16, and LCDR3 as shown in SEQ ID NO: 17. This antibody is designated herein as "BGB-A317" or "tislelizumab" and the sequence of this antibody is shown in Table 2. Table 2 Further changes to the Fc region framework

[0066] In other aspects, the Fc region is changed by replacing at least one amino acid residue with a different amino acid residue to change the effector function of the antibody. For example, one or more amino acids can be replaced with different amino acid residues so that the antibody has a changed affinity for the effector ligand, but retains the antigen binding ability of the parent antibody. The effector ligand with changed affinity can be, for example, an Fc receptor or the C1 component of complement. This method is described in, for example, U.S. Patent Nos. 5,624,821 and 5,648,260 by Winter et al.

[0067] In another aspect, one or more amino acid residues can be replaced with one or more different amino acid residues such that the antibody has altered CIq binding and / or reduced or abolished complement dependent cytotoxicity (CDC).This approach is described, for example, in U.S. Patent No. 6,194,551 to Idusogie et al.

[0068] On the other hand, one or more amino acid residues are changed to change the ability of the antibody to fix complement. This method is described in, for example, PCT Publication WO 94 / 29351 of Bodmer et al. In a specific aspect, one or more amino acids of the antibody or its antigen-binding fragment of the present disclosure are replaced by one or more allotype amino acid residues of IgG1 subclass and κ isotype. Allotype amino acid residues also include but are not limited to the heavy chain constant region of IgG1, IgG2 and IgG3 subclasses and the light chain constant region of κ isotype, as described in Jefferis et al., MAbs [monoclonal antibodies] 1: 332-338 (2009).

[0069] In another aspect, the Fc region is modified by modifying one or more amino acids to increase the ability of the antibody to mediate antibody-dependent cellular cytotoxicity (ADCC) and / or to increase the affinity of the antibody for Fcγ receptors. This approach is described, for example, in PCT publication WO 00 / 42072 by Presta. In addition, the binding sites for FcγRI, FcγRII, FcγRIII, and FcRn on human IgG1 have been mapped, and variants with improved binding have been described (see Shields et al., J. Biol. Chem. [Journal of Biological Chemistry] 276: 6591-6604, 2001).

[0070] In yet another aspect, the glycosylation of the modified antibody. For example, an aglycosylated antibody (i.e., the antibody lacks or has reduced glycosylation) can be prepared. Glycosylation can be changed, for example, to increase the affinity of the antibody to the "antigen". This carbohydrate modification can be achieved by, for example, changing one or more glycosylation sites within the antibody sequence. For example, one or more amino acid substitutions can be made, which result in the elimination of one or more variable region framework glycosylation sites, thereby eliminating the glycosylation of the site. This aglycosylation can increase the affinity of the antibody to the antigen. This method is described in, for example, U.S. Patent Nos. 5,714,350 and 6,350,861 of Co et al.

[0071] Additionally or alternatively, antibodies with altered glycosylation types can be prepared, such as hypofucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisecting GlcNac structures. It has been shown that this altered glycosylation pattern increases the ADCC ability of the antibody. This carbohydrate modification can be achieved, for example, by expressing the antibody in a host cell with an altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells in which recombinant antibodies are expressed to produce antibodies with altered glycosylation. For example, EP 1,176,195 by Hang et al. describes a cell line with a functionally disrupted FUT8 gene that encodes a fucosyltransferase, such that antibodies expressed in this cell line exhibit hypofucosylation. PCT Publication WO 03 / 035835 to Presta describes a variant CHO cell line, Lec13 cells, which have a reduced ability to attach fucose to Asn(297)-linked carbohydrates, also resulting in hypofucosylation of antibodies expressed in the host cells (see also Shields et al., (2002) J. Biol. Chem. 277:26733-26740). PCT Publication WO 99 / 54342 to Umana et al. describes cell lines that are engineered to express glycoprotein modifying glycosyltransferases (e.g., β(1,4)-N-acetylglucosaminyltransferase III (GnTIII)) such that antibodies expressed in the engineered cell lines exhibit increased bisecting GlcNac structures, which results in increased ADCC activity of the antibodies (see also Umana et al., Nat. Biotech. 17:176-180, 1999).

[0072] On the other hand, if a reduction in ADCC is desired, many previous reports show that human antibody subclass IgG4 has only moderate ADCC and almost no CDC effector function (Moore GL et al. 2010 MAbs [monoclonal antibodies], 2: 181-189). On the other hand, native IgG4 was found to be less stable under stress conditions (such as in acidic buffers or at elevated temperatures) (Angal, S. 1993 Mol Immunol [molecular immunology], 30: 105-108; Dall'Acqua, W. et al., 1998 Biochemistry [biochemistry], 37: 9266-9273; Aalberse et al., 2002 Immunol [immunology], 105: 9-19). Reduced ADCC can be achieved by operably linking the antibody to an IgG4 engineered with a combination of alterations having reduced or ineffective FcγR binding or C1q binding activity, thereby reducing or eliminating ADCC and CDC effector functions. Considering the physicochemical properties of antibodies as biopharmaceuticals, one of the less desirable inherent properties of IgG4 is that its two heavy chains dynamically separate in solution to form half antibodies, which leads to the generation of bispecific antibodies in vivo through a process called "Fab arm exchange" (Van der Neut Kolfschoten M et al. 2007 Science, 317: 1554-157). The mutation of serine at position 228 (EU numbering system) to proline exhibits an inhibitory effect on the separation of IgG4 heavy chains (Angal, S. 1993 Mol Immunol, 30: 105-108; Aalberse et al. 2002 Immunol, 105: 9-19). It has been reported that some amino acid residues in the hinge region and the γFc region have an influence on the interaction of antibodies with Fcγ receptors (Chappel SM et al., 1991 Proc. Natl. Acad. Sci. USA [Proceedings of the National Academy of Sciences of the United States of America], 88:9036-9040; Mukherjee, J. et al., 1995 FASEB J [Journal of the Federation of American Societies for Experimental Biology], 9:115-119; Armour, KL et al., 1999 Eur J Immunol [European Journal of Immunology], 29:2613-2624; Clynes, RA et al., 2000 Nature Medicine [Nature Medicine], 6:443-446; Arnold JN, 2007 Annu Rev immunol [Annual Review of Immunology], 25:21-50).In addition, some rare IgG4 isotypes in the human population may also give rise to different physicochemical properties (Brusco, A. et al., 1998 Eur J Immunogenet [European Journal of Immunogenetics], 25:349-55; Aalberse et al., 2002 Immunol [Immunology], 105:9-19). Antibody production

[0073] Osperizumab and tislelizumab antibodies and antigen-binding fragments thereof can be produced by any method known in the art, including but not limited to recombinant expression of antibody tetramers, chemical synthesis and enzymatic digestion, while full-length monoclonal antibodies can be obtained, for example, by hybridoma or recombinant production. Recombinant expression can be from any suitable host cell known in the art, such as mammalian host cells, bacterial host cells, yeast host cells, insect host cells, etc.

[0074] The disclosure also provides polynucleotides encoding antibodies described herein, such as polynucleotides encoding heavy or light chain variable regions or segments comprising complementary determining regions described herein. In some aspects, the polynucleotides encoding the heavy chain variable region have at least 85%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% nucleic acid sequence identity with the polynucleotides encoding the polypeptides of Table 1 or Table 2. In some aspects, the polynucleotides encoding the light chain variable region have at least 85%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% nucleic acid sequence identity with the polynucleotides encoding the polypeptides of Table 1 or Table 2.

[0075] The present disclosure also provides expression vectors and host cells for producing osperlizumab and / or tislelizumab antibodies. The selection of expression vectors depends on the expected host cell of the expression vector. Typically, the expression vector contains promoters and other regulatory sequences (e.g., enhancers) that are operably connected to polynucleotides encoding osperlizumab and / or tislelizumab antibody chains or antigen-binding fragments. In some aspects, in addition to under the control of inducing conditions, inducible promoters are used to prevent the expression of inserted sequences. Inducible promoters include, for example, arabinose, lacZ, metallothionein promoters or heat shock promoters. The culture of transformed organisms can be expanded under non-inducing conditions, but not in the case of a population biased towards the host cell to better tolerate the coding sequence of its expression product. In addition to promoters, other regulatory elements may also be required or desired for the effective expression of osperlizumab and / or tislelizumab antibodies or antigen-binding fragments. These elements typically include an ATG start codon and adjacent ribosome binding sites or other sequences. In addition, expression efficiency can be increased by including an enhancer appropriate for the cell system in use (see, e.g., Scharf et al., Results Probl. Cell Differ. 20:125, 1994; and Bittner et al., Meth. Enzymol. 153:516, 1987). For example, the SV40 enhancer or the CMV enhancer can be used to increase expression in mammalian host cells.

[0076] The host cell for carrying and expressing the antibody chain of osperlizumab and / or tislelizumab can be prokaryotic or eukaryotic. Escherichia coli is a prokaryotic host that can be used to clone and express the polynucleotides disclosed herein. Other applicable microbial hosts include bacilli, such as Bacillus subtilis, and other Enterobacteriaceae, such as Salmonella, Serratia and various Pseudomonas. In these prokaryotic hosts, expression vectors can also be prepared, which typically contain expression control sequences (e.g., replication origins) compatible with the host cells. In addition, there will be any number of well-known promoters, such as lactose promoter systems, tryptophan (trp) promoter systems, β-lactamase promoter systems, or promoter systems from bacteriophage λ. Promoters typically optionally control expression with operator sequences, and have ribosome binding site sequences, etc., for initiating and completing transcription and translation. Other microorganisms such as yeast can also be used to express osperlizumab and / or tislelizumab polypeptides. A combination of insect cells and baculovirus vectors can also be used.

[0077] In other aspects, mammalian host cells are used to express and produce the osperlimab and / or tislelizumab polypeptides disclosed herein. For example, they can be hybridoma cell lines expressing endogenous immunoglobulin genes or mammalian cell lines carrying exogenous expression vectors. These include any normal mortal or normal or abnormal immortal animal or human cells. For example, many suitable host cell lines capable of secreting complete immunoglobulins have been developed, including CHO cell lines, various COS cell lines, HEK 293 cells, myeloma cell lines, transformed B cells and hybridomas. The use of mammalian tissue cell cultures to express polypeptides is generally discussed in, for example, Winnacker, From Genes to Clones, VCH Press, NY, NY, 1987. Expression vectors for mammalian host cells may include expression control sequences, such as replication origins, promoters and enhancers (see, e.g., Queen et al., Immunol. Rev. [Immunology Review] 89: 49-68, 1986) and necessary processing information sites, such as ribosome binding sites, RNA splicing sites, polyadenylation sites and transcription terminator sequences. These expression vectors typically contain promoters derived from mammalian genes or mammalian viruses. Suitable promoters may be constitutive, cell type specific, stage specific, and / or regulatable or adjustable. Useful promoters include, but are not limited to, metallothionein promoters, constitutive adenovirus major late promoters, dexamethasone-inducible MMTV promoters, SV40 promoters, MRP polIII promoters, constitutive MPSV promoters, tetracycline-inducible CMV promoters (e.g., human immediate early CMV promoters), constitutive CMV promoters, and promoter-enhancer combinations known in the art. Testing and diagnostic methods

[0078] The antibodies or antigen-binding fragments disclosed herein can be used in a variety of applications, including but not limited to methods for detecting TIGIT. In one aspect, the antibodies or antigen-binding fragments can be used to detect the presence of TIGIT in a biological sample. As used herein, the term "detection" includes quantitative or qualitative detection. In some aspects, the biological sample includes cells or tissues. In other aspects, these tissues include normal and / or cancerous tissues that express TIGIT at higher levels relative to other tissues.

[0079] In one aspect, the present disclosure provides a method for detecting the presence of TIGIT in a biological sample. In some aspects, the method includes contacting the biological sample with an anti-TIGIT antibody under conditions that allow the antibody to bind to the antigen, and detecting whether a complex is formed between the antibody and the antigen. The biological sample may include, but is not limited to, a urine or blood sample.

[0080] Also included is a method for diagnosing an obstacle associated with TIGIT expression. In some aspects, the method includes contacting a test cell with an anti-TIGIT antibody; determining the expression level of TIGIT in the test cell (quantitatively or qualitatively) by detecting the binding of the anti-TIGIT antibody to the TIGIT polypeptide; and comparing the expression level of the test cell with the TIGIT expression level in a control cell (e.g., a normal cell or non-TIGIT expressing cell of the same tissue source as the test cell), wherein a higher level of TIGIT expression in the test cell compared to the control cell indicates the presence of an obstacle associated with TIGIT expression. Treatment

[0081] The antibodies or antigen-binding fragments of the present disclosure can be used in a variety of applications, including but not limited to methods for treating TIGIT-related disorders or diseases. In one aspect, the TIGIT-related disorder or disease is lymphoma and in certain aspects, is DLBCL or refractory or relapsed DLBCL.

[0082] In one aspect, the present disclosure provides a method for treating cancer. In some aspects, the method comprises administering an effective amount of an anti-TIGIT antibody or antigen-binding fragment to a patient in need. The cancer may include, but is not limited to, lymphoma, leukemia, DLBCL, and refractory or recurrent DLBCL.

[0083] Antibodies or antigen-binding fragments of the present invention can be administered by any suitable means, including parenteral, intrapulmonary and intranasal, and if necessary for local treatment, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. Administration can be by any suitable route, for example by injection, such as intravenous or subcutaneous injection, which depends in part on whether the administration is short-lived or long-term. Various dosing regimens are contemplated herein, including but not limited to single administration or multiple administrations at different time points, push injection administration and pulse infusion.

[0084] The antibody or antigen-binding fragment of the present invention can be formulated, administered and used in a manner consistent with good medical practice. Factors to be considered in this regard include the specific obstacle for treatment, the specific mammal for treatment, the clinical condition of the individual patient, the cause of the obstacle, the delivery site of the agent, the method of administration, the administration regimen and other factors known to medical practitioners. The antibody does not need to be formulated with one or more agents currently used to prevent or treat the obstacle under study. The effective amount of such other agents depends on the amount of the antibody present in the formulation, the type of obstacle or treatment and other factors discussed above. These are usually used with the same dosage and route of administration as described herein, or with about 1%-99% of the dosage described herein, or with any dosage and any route determined to be suitable by experience / clinical use.

[0085] For the prevention or treatment of disease, the appropriate dosage of the antibody or antigen-binding fragment of the present invention will depend on the type of disease to be treated, the type of antibody, the severity and course of the disease, whether the antibody is used for prevention or treatment purposes, previous therapy, the patient's clinical history and response to the antibody, and the judgment of the attending physician. The antibody is appropriately administered to the patient once or through a series of treatments. Depending on the type and severity of the disease, an antibody of about 1 μg / kg to 100 mg / kg can be an initial candidate dose for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. Depending on the above factors, a typical daily dose can be about 1 μg / kg to 100 mg / kg or more. For repeated administration over a few days or longer, depending on the condition, treatment will generally continue until the desired suppression of disease symptoms occurs. Such doses can be administered intermittently, for example, weekly or every three weeks (for example, so that the patient receives about two to about twenty, or for example, about six doses of the antibody). An initial higher loading dose is administered, followed by one or more lower doses. However, other dosing regimens may be useful. The progress of the therapy can be easily monitored by conventional techniques and assays. Combination therapy

[0086] In one aspect, the osperlimab antibody can be used in combination with other therapeutic agents (e.g., anti-PD1 antibodies). Anti-PD1 antibodies may include, but are not limited to, tislelizumab as described above. Pembrolizumab (formerly known as MK-3475) disclosed by Merck is a humanized IgG4-K immunoglobulin with a molecular weight of approximately 149 kDa that targets the PD1 receptor and inhibits the binding of the PD1 receptor ligand PD-L1 to PD-L2. Pembrolizumab has been approved for indications of metastatic melanoma and metastatic non-small cell lung cancer (NSCLC), and is in clinical studies for the treatment of head and neck squamous cell carcinoma (HNSCC) and refractory Hodgkin lymphoma (cHL). Nivolumab (as disclosed by Bristol-Meyers Squibb) is a fully human IgG4-K monoclonal antibody. Nivolumab (clone 5C4) is disclosed in U.S. Pat. No. US 8,008,449 and WO 2006 / 121168. Nivolumab is approved for the treatment of melanoma, lung cancer, kidney cancer, and Hodgkin lymphoma.

[0087] On the other hand, osperlimab antibodies can be used with anti-CD20 antibodies, which may include, but are not limited to, rituximab (Rituxan), ibritumomab tiuxetan (Zevalin), tositumomab (Bexxar), and obinutuzumab (Gazyva). Anti-CD20 antibodies (e.g., rituximab) are directed against the CD20 antigen expressed on the surface of pre-B and mature B lymphocytes. After binding to CD20, these antibodies mediate B cell lysis, with possible mechanisms of lysis including complement-dependent cytotoxicity (CDC) and ADCC. In NHL clinical trials, the most common adverse events associated with rituximab (≥25%) were infusion reactions, fever, lymphopenia, chills, infections, and fatigue. Tumor lysis syndrome (TLS) may be associated with rituximab treatment in patients with NHL with high numbers of circulating malignant cells (≥25,000 / mm 3 ) or higher tumor burden, leading to a higher risk of TLS. Rituximab is currently approved for the treatment of CD20 + NHL, including CD20 + DLBCL.

[0088] Another combination for treating DLBCL includes administering osperlimab, tislelizumab and / or rituximab together with current DLBCL treatment. Adding rituximab (R-CHOP) to the chemotherapy regimen of cyclophosphamide, doxorubicin, vincristine and prednisone significantly improved clinical outcomes (Coiffier et al., N Engl J Med. [New England Journal of Medicine] 2002; 346 (4): 235-242). Recently, the Phase III POLARIX study compared the combination of polatuzumab vedotin with the chemotherapy regimen R-CHP and with the combination of standard care R-CHOP for first-line DLBCL, and this study reached the primary endpoint, demonstrating significantly improved and clinically meaningful progression-free survival. However, 30% to 40% of DLBCL patients will eventually relapse, of which 10% are primary refractory, which remains an ongoing clinical problem (Flowers et al., CA Cancer J Clin. 2010; 60(6):393-408). Therefore, the combination of osperlimab, tislelizumab and / or rituximab with the administration of chemotherapy may prove useful for the treatment of DLBCL and refractory or relapsed DLBCL. Pharmaceutical compositions and formulations

[0089] Also provided are compositions comprising anti-TIGIT antibodies or antigen-binding fragments, or polynucleotides comprising sequences encoding anti-TIGIT antibodies or antigen-binding fragments, including pharmaceutical formulations. In certain embodiments, the composition comprises one or more antibodies or antigen-binding fragments that bind to TIGIT, or one or more polynucleotides comprising sequences encoding one or more antibodies or antigen-binding fragments that bind to TIGIT. These compositions may also include suitable carriers, such as pharmaceutically acceptable excipients well known in the art, including buffers.

[0090] Pharmaceutical formulations of the TIGIT antibodies or antigen-binding fragments described herein are prepared by mixing such antibodies or antigen-binding fragments having the desired purity with one or more optional pharmaceutically acceptable carriers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. ed. (1980)) in the form of a lyophilized formulation or an aqueous solution. Pharmaceutically acceptable carriers are generally non-toxic to recipients at the doses and concentrations employed, and include, but are not limited to: buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens such as methyl paraben or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than 1% to 1% of the total weight of the drug); about 10 residues); proteins, such as serum albumin, gelatin or immunoglobulin; hydrophilic polymers, such as polyvinyl pyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrin; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions, such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, such as polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers herein also include interstitial drug dispersants, such as soluble neutral active hyaluronidase glycoprotein (sHASEGP), such as human soluble PH-20 hyaluronidase glycoprotein, such as rHuPH20 ( Baxter International, Inc. Certain exemplary sHASEGPs and methods of use, including rHuPH20, are described in U.S. Pat. Nos. 7,871,607 and 2006 / 0104968. In one aspect, sHASEGP is combined with one or more additional glycosaminoglycanases such as chondroitinase.

[0091] Exemplary lyophilized antibody formulations are described in US Pat. No. 6,267,958. Aqueous antibody formulations include those described in US Pat. No. 6,171,586 and WO 2006 / 044908, the latter including a histidine-acetate buffer.

[0092] Sustained release preparations may be prepared. Suitable examples of sustained release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, eg films, or microcapsules.

[0093] Formulations for in vivo administration are generally sterile. Sterility can be readily achieved, for example, by filtration through sterile filtration membranes. Pharmaceutical compositions and kits

[0094] In some aspects, the present disclosure provides a composition, such as a pharmaceutically acceptable composition, comprising an anti-TIGIT antibody as described herein formulated with at least one pharmaceutically acceptable excipient. As used herein, the term "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, isotonic agents, and absorption delaying agents that are physiologically compatible, etc. The excipient may be suitable for intravenous, intramuscular, subcutaneous, parenteral, rectal, spinal or epidermal administration (e.g., by injection or infusion).

[0095] The compositions herein can be in various forms. These include, for example, liquid, semisolid and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, liposomes and suppositories. The appropriate form depends on the intended mode of administration and therapeutic application. Typical suitable compositions are in the form of injectable or infusible solutions. A suitable mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In certain embodiments, the antibody is administered by intravenous infusion or injection. In certain embodiments, the antibody is administered by intramuscular or subcutaneous injection. Examples Example 1: Osperizumab and tislelizumab in the treatment of DLBCL

[0096] TIGIT and PD1 act as immune checkpoint receptors in the overlapping regulation of immune tolerance. It has been shown that blocking TIGIT receptors alone or in combination with PD1 / PD-L1 blocking can rescue functionally "exhausted" T cells in vitro and in vivo (Chauvin et al., J Clin Invest. [Clinical Research Journal] 2015; 125: 2046-58). In mouse models, the combination of TIGIT blocking and anti-PD1 / PD-L1 antibodies showed significantly better anti-tumor efficacy than any single therapy (Dixon et al., J Immunol. [Immunology Journal] 2018; 200: 3000-7). Clinically, treatment with a combination of osperlimab and tislelizumab in DLBCL that has not previously been treated with checkpoint inhibitor therapy may be more effective than tislelizumab alone.

[0097] For example, in first-line stage IV NSCLC patients with a PD-L1 tumor proportion score ≥1%, the combination of tirilimumab plus atezolizumab reported an overall response rate of 31% and a progression-free survival of 5.42 months, while the overall response rate of placebo plus atezolizumab was 16% and the progression-free survival was 3.58 months (Rodriguez-Abreu et al., J Clin Oncol. 2020;38(Suppl 15):9503). Similarly, in patients with NSCLC who had not received treatment or had received at least 1 line of platinum-containing chemotherapy and had not received prior PD1 / PD-L1 therapy, the combination of vembrolizumab plus pembrolizumab reported an unconfirmed and confirmed ORR of 29% and 24%, respectively (Niu et al., Annals of Oncology 2020;31:S891-2).

[0098] Gene expression analysis has previously demonstrated upregulation of TIGIT and PD-L1 in DLBCL compared with normal controls (Laurent et al., 2015; 4(8): e1026530). Characterization of co-inhibitory receptor expression in DLBCL intratumoral T cells revealed that both TIGIT and PD1 were expressed more frequently than all other receptors (such as TIM-3 and LAG-3). The co-expression pattern of TIGIT and PD1 not only resulted in impaired T cell effector function (as tested by cytokine production), but also inhibited the antitumor activity of T cells by interacting with tumor cells expressing ligands in DLBCL (CD155 and CD122 expression). All of this suggests the value of combined blockade of TIGIT and PD1 in DLBCL and in exploring immune escape mechanisms by characterizing the tumor microenvironment (Josefsson et al., Cancer Immunol Res. [Cancer Immunology Research] 2019; 7(3): 355-362).

[0099] However, limitations of anti-PD1 single agents have also been observed in patients with R / R DLBCL, with an overall response rate of approximately 10%. In the combination therapy of nivolumab and ibrutinib, an increasing trend in overall response rate and complete response rate was observed in patients with high expression of PD-L1 on the tumor cell surface. Given the tolerable and manageable safety profile of tislelizumab in DLBCL and the scientific basis for the synergistic antitumor effect between anti-TIGIT and anti-PD1 therapy, the combination of osperlimab and tislelizumab may bring significant clinical benefit to R / R DLBCL in patients with PD-L1-positive disease and support further clinical development.

[0100] Safety and efficacy data for tislelizumab in NHL, including DLBCL, are available from an ongoing study of the combination therapy. A Phase 1b, open-label, multiple-dose, dose-escalation and dose-expansion study of zanubrutinib in combination with tislelizumab in patients with B-cell lymphoid malignancies (i.e., NHL) is currently ongoing.

[0020] For patients who are PD-L1 positive, they will be assigned to Cohort 1 - Cohort 1: DLBCL or R / R DLBCL patients with PD-L1 positive (any expression level) on the tumor cell surface will receive the combination of ospelizumab and tislelizumab. Cohort 1 will enroll approximately 43 to 50 patients. Patients will receive 21-day treatment cycles. The combination of ospelizumab and tislelizumab will be administered continuously intravenously on Day 1 of each 21-day cycle. The combination of ospelizumab 900 mg and tislelizumab 200 mg will be administered on Day 1 of each 3-week cycle. Example 2: Osperlimab and Rituximab in the treatment of DLBCL

[0021] To ensure the greatest clinical benefit in patients with PD-L1-negative DLBCL, the combination of rituximab and ospelimab will be administered in patients with DLBCL or refractory / relapsed (R / R) DLBCL. Ospelimab works synergistically with rituximab to promote immediate tumor killing and long-term memory immune responses by enhancing T cell / NK cell activation and Treg depletion / suppression.

[0022] This study will determine the preliminary antitumor activity of ospelizumab in combination with either tislelizumab or rituximab in patients with DLBCL or R / RDLBCL. Figure 1). As mentioned above, eligible patients will be assigned to 2 cohorts based on the level of PD-L1 expression on the surface of tumor cells, which can be tested by immunohistochemistry (IHC) in local laboratories. Those patients who are PD-L1 negative will be assigned to cohort 2- Cohort 2: DLBCL or R / R DLBCL patients whose tumor cells are PD-L1 negative will receive the combination of ospelimab and rituximab.

[0023] Approximately 23 to 30 patients in Cohort 2 will be included in the entire study. Each treatment cycle consists of 21 days. The combination of osperlimab and rituximab in Cohort 2 will be administered continuously intravenously on Day 1 of each 21-day cycle. Osperlimab will initially be tested at a dose level of 900 mg. If osperlimab 900 mg exceeds the MTD, osperlimab 600 mg will be tested. Other dose levels of osperlimab (e.g., less than 600 mg) may be explored to confirm the optimal dose level of osperlimab in combination with rituximab. The administered dose of rituximab will be 375 mg / m 2 . Example 3: Osperizumab, tislelizumab, and rituximab in the treatment of DLBCL

[0020] For patients with PD-L1-positive DLBCL or R / R DLBCL, administration of osperlimab, tislelizumab, rituximab, and chemotherapy will be explored. TIGIT blockade by osperlimab has the potential to promote NK cell activation against poliovirus receptor+ (PVR+) tumor cells and synergize ADCC activity mediated by therapeutic monoclonal antibodies such as rituximab. Combining this with the ability of tislelizumab to rescue functionally "exhausted" T cells and chemotherapy indicated for R-CHOP (cyclophosphamide, hydroxydoxorubicin hydrochloride (doxorubicin hydrochloride), vincristine (Oncovin), and prednisone), the combination may resolve aggressive forms of DLBCL or R / R DLBCL.

Claims

1. A method for treating diffuse large B-cell lymphoma (DLBCL), the method comprising administering to a subject an effective amount of an anti-TIGIT antibody or an antigen-binding fragment thereof in combination with an effective amount of an anti-PD1 antibody or an antigen-binding fragment thereof.

2. The method of claim 1, wherein the method comprises administering to a subject an effective amount of an antibody or an antigen-binding fragment thereof, which specifically binds to human TIGIT and comprises: a heavy chain variable region comprising HCDR (heavy chain complementarity determining region) 1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3; and a light chain variable region comprising LCDR (light chain complementarity determining region) 1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO:

6.

3. The method of claim 2, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises: a heavy chain variable region (VH) comprising SEQ ID NO: 7 and a light chain variable region (VL) comprising SEQ ID NO:

8.

4. The method of claim 1, wherein the anti-PD1 antibody comprises an antibody or an antigen-binding fragment thereof, which specifically binds to human PD1 and comprises: A heavy chain variable region comprising: HCDR1 of SEQ ID NO:12, HCDR2 of SEQ ID NO:13, and HCDR3 of SEQ ID NO:14; and a light chain variable region comprising LCDR1 of SEQ ID NO:15, LCDR2 of SEQ ID NO:16, and LCDR3 of SEQ ID NO:

17.

5. The method of claim 4, wherein the anti-PD1 antibody or antigen-binding fragment thereof specifically binds to human PD1 and comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 18 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:

19.

6. The method of claim 4 or 5, wherein the anti-PD1 antibody comprises an IgG4 constant domain comprising SEQ ID NO:

20.

7. The method of claim 1, wherein the anti-TIGIT antibody is an antibody fragment selected from the group consisting of: Fab, Fab'-SH, Fv, scFv, and (Fab')2 fragment.

8. The method of claim 1, wherein the anti-PD1 antibody is an antibody fragment selected from the group consisting of: Fab, Fab'-SH, Fv, scFv, and (Fab')2 fragment.

9. A method for treating diffuse large B-cell lymphoma (DLBCL), the method comprising administering to a subject an effective amount of an anti-TIGIT antibody or an antigen-binding fragment thereof in combination with an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof.

10. The method of claim 9, wherein the method comprises administering to a subject an effective amount of an antibody or an antigen-binding fragment thereof, which specifically binds to human TIGIT and comprises: a heavy chain variable region comprising HCDR (heavy chain complementarity determining region) 1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3; and a light chain variable region comprising LCDR (light chain complementarity determining region) 1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO:

6. 11 . The method of claim 10 , wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises: a heavy chain variable region (VH) comprising SEQ ID NO: 7 and a light chain variable region (VL) comprising SEQ ID NO:

8.

12. The method of claim 9, wherein the anti-CD20 antibody is rituximab (Rituxan), ibritumomab tiuxetan (Zevalin), tositumomab (Bexxar), or obinutuzumab (Gazyva).

13. The method of claim 12, wherein the anti-CD20 antibody is rituximab (Rituxan).

14. The method of claim 1 or claim 9, wherein the DLBCL is relapsed or refractory DLBCL.

15. The method of claim 1 or claim 9, further comprising administering chemotherapy.

16. The method of claim 15, wherein the chemotherapy is cyclophosphamide, doxorubicin, vincristine, and prednisone.

17. The method of claim 1, wherein the anti-PD1 antibody is administered at 200 mg every three weeks.

18. The method of claim 1, wherein the anti-TIGIT antibody is administered in the range of 50 mg-900 mg.

19. The method of claim 1, wherein the anti-TIGIT antibody is administered at 50 mg every three weeks.

20. The method of claim 1, wherein the anti-TIGIT antibody is administered at 150 mg every three weeks.

21. The method of claim 1, wherein the anti-TIGIT antibody is administered at 450 mg every three weeks.

22. The method of claim 1, wherein the anti-TIGIT antibody is administered at 900 mg every three weeks.

23. The method of claim 13, wherein Rituxan is administered at 375 mg / m 2 Application.

24. A method for treating diffuse large B-cell lymphoma (DLBCL), the method comprising administering to a subject an effective amount of an anti-TIGIT antibody or an antigen-binding fragment thereof in combination with an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof and an effective amount of an anti-PD1 antibody.

25. The method of claim 24, wherein the method comprises administering to the subject an effective amount of an antibody or an antigen-binding fragment thereof, which antibody or antigen-binding fragment thereof specifically binds to human TIGIT and comprises: a heavy chain variable region comprising HCDR (heavy chain complementarity determining region) 1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3; and a light chain variable region comprising LCDR (light chain complementarity determining region) 1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO:

6.

26. The method of claim 25, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises: a heavy chain variable region (VH) comprising SEQ ID NO: 7 and a light chain variable region (VL) comprising SEQ ID NO:

8.

27. The method of claim 24, wherein the anti-PD1 antibody comprises an antibody or an antigen-binding fragment thereof that specifically binds to human PD1 and comprises: A heavy chain variable region comprising: HCDR1 of SEQ ID NO:12, HCDR2 of SEQ ID NO:13, and HCDR3 of SEQ ID NO:14; and a light chain variable region comprising LCDR1 of SEQ ID NO:15, LCDR2 of SEQ ID NO:16, and LCDR3 of SEQ ID NO:

17.

28. The method of claim 27, wherein the anti-PD1 antibody or antigen-binding fragment thereof specifically binds to human PD1 and comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 18 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:

19.

29. The method of claim 24, further comprising administering chemotherapy.

30. The method of claim 29, wherein the chemotherapy is cyclophosphamide, doxorubicin, vincristine, and prednisone.

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