Immunotherapeutic methods for urothelial carcinoma

Abstract

The present invention discloses a method of immunotherapy for a patient with urothelial carcinoma (UC), the method comprising administering to the patient an anti-PD-1 antibody that reduces antibody dependent phagocytosis by reducing Fc [gamma] R binding, and a method of immunotherapy for a patient with urothelial carcinoma (UC), the method comprising administering to the patient an anti-PD-1 antibody that reduces antibody dependent phagocytosis by reducing Fc [gamma] R binding.

Description

This application is a divisional application of the invention application with the application date of February 8, 2019, Chinese application number 201980024421.9, and invention name “Immunotherapy method for urothelial carcinoma”. CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. US 62 / 628,648, filed on February 9, 2018, and International Application No. PCT / CN 2018 / 076100, filed on February 10, 2018, and the entire contents of which are incorporated herein by reference. Instructions for Electronic Submission of Text Files

[0002] All citations of the electronically submitted text files of this application are incorporated into this application: Sequence Listing in computer-readable form (file name: BEIG_034_01WO_SeqList.TXT, data recorded on February 8, 2019, file size 126 kilobytes). Technical Field

[0003] Disclosed herein is a treatment method for patients with urothelial carcinoma (UC), comprising administering to the patient an anti-PD-1 antibody that is specifically engineered to reduce FcγR binding to macrophages, thereby abrogating antibody-dependent phagocytosis. Background Art

[0004] Urothelial carcinoma is also known as urothelial (transitional cell) carcinoma (UC) or urothelial carcinoma (transitional cell tumor, or transitional cell tumor of the urothelium), as well as bladder cancer. More than 90% of urothelial tumors arise from the bladder, 8% from the renal pelvis, and the remaining 2% from the ureters and urethra.

[0005] Platinum-based combination chemotherapy has been used as a first-line therapy for patients with advanced urothelial carcinoma for decades. For example, the standard of care for first-line bladder cancer treatment includes a combination of methotrexate, vincristine, doxorubicin, and cisplatin (MVAC), which has been shown to have considerable toxicity. Another combination chemotherapy is cisplatin-gemcitabine, which has replaced MVAC as the first-line standard of care for metastatic bladder cancer, and it is reported that 90% of patients have relapsed and have a very poor prognosis. Progress in the treatment of metastatic UC has stagnated over the past three decades.

[0006] WO 2016064649 A1 discloses a combination therapy of the anti-VEGF receptor 2 antibody ramucirumab and docetaxel for the simultaneous, separate, or sequential treatment of UC patients. This suggests that antibody therapy can be an effective drug for treating urothelial carcinoma.

[0007] Monoclonal antibodies (mAbs) directed against immune checkpoint inhibitory receptors, such as programmed cell death-1 (PD-1), have demonstrated promising antitumor activity in a variety of malignancies (Topalian SL, et al. N Engl J Med. 2012;366:2443-2454.), including UC (Plimack ER, et al. J Clin Oncol. 2015;33 (Suppl; Abstract 4502); Bellmunt J, et al. Abstract 470. Presented at the 31st Annual Meeting of the Society for Immunotherapy of Cancer, November 9-13, 2016, National Harbor, MD, USA; Balar A, et al. Ann Oncol. 2016; Vol. 27, Suppl_6, Oct 1, 2016, LBA32 PR.; Sharma P, et al. J Clin Oncol. 2016;34(Suppl ab 4501); Galsky MD, et al. J Clin Oncol. 2016;LB:A31; and Sharma P, et al. Lancet Oncol. 2016;17:1590-1598).

[0008] PD-1 in CD8 + Effector, tumor-infiltrating T lymphocytes (TILs) are relatively overexpressed; and anti-PD-1 antibodies induce CD8 + Increase in the percentage of T cells (Ahmadzadeh M, et al. Blood. 2009; 114: 1537-1544).

[0009] WO 2015 / 035606 A1 discloses a humanized IgG4 monoclonal antibody. This monoclonal antibody has high affinity and binding specificity for PD-1, particularly specifically binding to PD-1 (including residues K45 and I93; or I93, L95, and P97) and inhibiting PD-1-mediated cell signaling in immune cells (through binding to a set of amino acid residues required for binding to its ligand PD-L1). WO 2015 / 035606 discloses a monoclonal antibody that binds to human PD-1 and a modified IgG4 Fc region, wherein the modified IgG4 Fc region reduces binding of the antibody to one or more Fcγ receptors.

[0010] Dahan R et al. reported in vivo evidence that anti-PD-1 antibodies showed reduced tumor cell cytotoxicity when the Fc domain of the antibody engaged the Fc-γ receptor (FcγR) (Dahan R, et al. Cancer Cell. 2015; 28: 285-295). + Preferential depletion of TILs. This reduction may be associated with reduced anti-PD-1 tumor activity.

[0011] The antibody of WO 2015 / 035606 A1 is specifically engineered to reduce FcγR binding to macrophages and myeloid-derived suppressor cells; reduction in FcγR binding may improve clinical activity by preserving activated T cells, while Figure 1 The proposed mechanism of action is shown graphically.

[0012] As shown by the advantages in the treatment of UC, the inventors of the present application have found that the antibodies of the present invention can be used as a new immunotherapy regimen for urothelial carcinoma including advanced or metastatic urothelial carcinoma. Summary of the Invention

[0013] The present application discloses a treatment method for patients with urothelial carcinoma (UC), comprising administering to the patient a therapeutically effective amount of an anti-PD-1 antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof is specifically engineered to reduce FcγR binding to macrophages, thereby reducing antibody-dependent phagocytosis.

[0014] In one embodiment, urothelial carcinoma involves the ureters, urethra, renal pelvis and / or bladder. In another embodiment, urothelial carcinoma is transitional cell carcinoma. In yet another embodiment, urothelial carcinoma is advanced or metastatic.

[0015] In one embodiment, the patient has PD-L1 + Urothelial carcinoma or PD-L1 -In another embodiment, the patient has PD-L1 + Urothelial carcinoma.

[0016] In one embodiment, the anti-PD-1 antibody is an antibody disclosed in WO 2015 / 035606 A1. The antibody disclosed in WO 2015 / 035606 A1 specifically binds to the programmed death-1 (PD-1) receptor and inhibits PD-1-mediated cell signaling in immune cells (by binding to a set of amino acid residues required for binding to its ligand PD-L1). In another embodiment, the anti-PD-1 antibody is a humanized monoclonal antibody comprising a heavy chain variable region (V H ) and a light chain variable region (V L ) (comprising SEQ ID No 24 and SEQ ID No 26, respectively) and an IgG4 heavy chain effector or constant domain (SEQ ID NO: 88), hereinafter referred to as Mab-1, which specifically binds to PD-1 (including PD-1 receptor residues K45 and I93; or I93, L95 and P97)) and inhibits PD-1-mediated cell signaling in immune cells.

[0017] In another embodiment, the anti-PD-1 antibody is a monoclonal antibody that binds to human PD-1 and comprises a modified IgG4 Fc region, wherein the modified IgG4 Fc region reduces binding of the antibody to one or more Fcγ receptors. In a preferred embodiment, the anti-PD-1 antibody is a monoclonal antibody that binds to human PD-1 and comprises an IgG4 Fc region comprising an S228P mutation at position 228 and amino acid mutations at positions 233, 234, and 235, wherein the mutations at positions 233, 234, and 235 cause the antibody to exhibit reduced binding to at least one Fcγ receptor relative to Fc binding of a reference IgG4 antibody having only the mutation at position 228 and no other Fc region mutations, as disclosed in WO 2015 / 035606 A1. In a further embodiment, the IgG4 Fc region comprises amino acid mutations at positions 228, 233, 234, 235, and 265, as disclosed in WO 2015 / 035606 A1. In a further embodiment, the IgG4 Fc region comprises amino acid mutations at positions 228, 233, 234, 235, 265, 309, and 409, as disclosed in WO 2015 / 035606 A1. In another embodiment, the IgG4 Fc region comprises amino acid mutations of S228P, E233P, F234V, and L235A, as disclosed in WO 2015 / 035606 A1. In one embodiment, the above-mentioned antibody may further comprise an IgG4 heavy chain constant domain comprising any one of SEQ ID NOs: 83-88.

[0018] The antibody claimed herein, Mab-1, has previously been shown to be tolerable in patients with advanced solid tumors such as hepatocellular carcinoma, and its toxicity profile indicates that adverse events (AEs) are generally low to moderate in severity, manageable, and reversible; Desai J, et al. J Immunother Cancer. 2016; 4(Suppl 1): P154; WO 2019 / 001417 published Jan 3, 2019).

[0019] The treatment methods disclosed herein have been shown to reduce, delay progression, and alleviate urothelial carcinoma in human patients. The inventors of the present invention have found that anti-PD-1 antibodies, such as Mab-1, can be used as an effective immunotherapy for the treatment of urothelial carcinoma, especially PD-L1 + Urothelial carcinoma, because of PD-L1 - Compared with UC, PD-L1 +The inventors also found that Mab-1 treatment was generally well tolerated in pre-treated patients with UC, and that adverse events reported in UC patients were consistent with the overall safety profile observed in the study and were generally low or moderate in severity, manageable, and reversible. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The potential mechanism of T cell clearance by the anti-PD-1 antibodies used in this application was shown to be that reduced FcγR binding prevented macrophage-mediated T cell clearance.

[0021] Figure 2 The design of this clinical study is shown.

[0022] Figure 3 The duration of treatment and response of UC patients treated with the antibodies of the invention are shown.

[0023] Figure 4 The greatest tumor reduction was shown in UC patients treated with the antibodies of the invention.

[0024] Figure 5 Shown are changes in target lesions over time in UC patients treated with the antibodies of the invention.

[0025] Figures 6A-6H are radiographic images of a 74-year-old male patient with PD-L1-positive UC who had previously failed three anticancer treatment regimens without any immunotherapy. The images show UC tumor regression from day 0 (baseline) to day 46 of the treatment cycle. DETAILED DESCRIPTION abbreviation

[0026] Throughout the detailed description and examples of the invention, the following abbreviations will be used: Ab antibody AE adverse event CD Cluster of Differentiation CDR complementarity determining region CR Full Response DPBS Dulbecco's phosphate-buffered saline FcγR Fc-γ receptor ip intraperitoneal or intraperitoneal iv intravenous or intravenous IFN-γ Interferon-γ IgG immunoglobulin G mAb monoclonal antibody max maximum value MHC major histocompatibility complex min minimum value NK natural killer cells po "orally" or "by mouth" PD progressive disease PD-1 Programmed death 1 protein, Pdcd-1, or CD279 PD-L1 Programmed cell death ligand-1 PDX patient-derived xenografts PR Partial Response Q2W Once every 2 weeks Q3W Once every 3 weeks Q4W Once every 4 weeks QW once a week RP2D Phase 2 recommended dose SD stable disease TILs tumor-infiltrating lymphocytes UC urothelial carcinoma V H Heavy chain variable region V L Light chain variable region definition

[0027] 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.

[0028] As used herein, including the appended claims, for example, “ one "," A sort of ” and “the” in the singular include their corresponding plural referents unless the context clearly indicates otherwise.

[0029] The term " Antibody " is used in the broadest sense and specifically covers antibodies (including full-length monoclonal antibodies) and antibody fragments so long as they recognize PD-1. Antibody molecules are generally monospecific but can also be described as heterospecific, heterospecific, or multispecific. Antibody molecules bind to a specific antigenic determinant or epitope on an antigen through a specific binding site.

[0030] In this article, the term " Monoclonal antibodies "or" mAbs"Monoclonal" refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules contained in the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include a plurality of different antibodies having different amino acid sequences in their variable domains, particularly their CDRs, which are generally specific for different epitopes. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. Monoclonal antibodies (mAbs) can be obtained by methods known to those skilled in the art. See, e.g., Kohler and Milstein, Nature (London) 256:495 (1975); U.S. Patent No. 4,376,110; Ausubel et al., Short Protocols in Molecular Biology, 3rd ed., Wiley & Sons, 1995; Harlow and Lane, Using Antibodies, A Laboratory Manual Manual [Using Antibodies: A Technical Experimental Guide] (1998); and Colligan et al., eds., Current Protocols in Immunology [Contemporary Immunology Program], Greene Publishing Assoc. and Wiley Interscience, New York, (1992, 1993). The mAbs disclosed herein can be of any immunoglobulin class, including IgG, IgM, IgD, IgE, IgA, and any subclass thereof. The hybridomas producing mAbs can be cultured in vitro or in vivo. High titer mAbs can be obtained in vivo by intraperitoneal injection of cells from a single hybridoma into mice, such as primary primed Balb / c mice, to produce ascites containing high concentrations of the desired mAb. MAbs of the IgM or IgG isotype can be purified from such ascites, or from the culture supernatant, using column chromatography methods well known to those skilled in the art.

[0031] Typically, the basic antibody structural unit comprises a tetramer. Each tetramer comprises two pairs of identical polypeptide chains, each pair having one "light chain" (about 25 kDa) and one "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 kappa and lambda light chains. In addition, human heavy chains are typically classified as alpha, delta, epsilon, gamma, or mu, and define the isotype of the antibody 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.

[0032] Each light chain / heavy chain (V L \V H ) pair of variable regions form the antibody binding site. Therefore, in general, an intact antibody has two binding sites. Except for bifunctional or bispecific antibodies, the two binding sites are generally the same.

[0033] Typically, the variable domains of heavy and light chains contain three hypervariable regions, also called " Complementarity determining regions (CDRs) ", which are located within relatively conserved framework regions (FRs). The CDRs are usually aligned by the framework regions, enabling binding to specific epitopes. In general, from N-terminus to C-terminus, both the light and heavy chain variable domains contain FR-1, CDR-1, FR-2, CDR-2, FR-3, CDR-3, and FR-4. In general, the amino acid assignments for each domain conform to the definition of protein sequences of immunological interest, as defined by 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.

[0034] the term" Hypervariable region"" refers to the amino acid residues in an antibody that are responsible for antigen binding. The hypervariable region comprises amino acid residues from the "complementarity determining regions" or "CDRs" (i.e., CDR-L1, CDR-L2, and CDR-L3 in the light chain variable domain and CDR-H1, CDR-H2, and CDR-H3 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. Health [National Institutes of Health], Bethesda, MD (defining the CDR regions of antibodies by sequence); see also Chothia and Lesk (1987) J. Mol. Biol. [Journal of Molecular Biology] 196:901-917 (defining the CDR regions of antibodies by structure). The term "framework" or "FR" residues refers to those variable domain residues other than the hypervariable region residues defined herein as CDR residues. The definition of antigen binding sites is described in the following references: Ruiz et al., IMGT, International ImMunoGeneTics Database. Nucleic Acid Res. [Nucleic Acids Research], 28, 219-221 (2000); and Lefranc, M.-P. IMGT, International ImMunoGeneTics Database. Nucleic Acid Res. [Nucleic Acids Research], January 1; 29(1):207-09 (2001); MacCallum et al., Antibody-antigen interaction: Contact analysis and binding site topography [Antibody-antigen interactions: binding analysis and binding site distribution], J Mol. Biol., 262(5), 732-745 (1996); and Martin et al., Proc. Natl Acad. Sci. USA, 86, 9268-9272 (1989); Martin, et al., Methods Enzymol., 203, 121-153, (1991); Pedersen et al., Immunomethods, 1, 126, (1992); and Rees et al., in Sternberg MJE (ed.), Protein Structure Prediction. Oxford University Press, Oxford, 141-172 1996).

[0035] Unless otherwise stated, " Antibody fragments "or" antigen-binding fragment " refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to specifically bind to the antigen bound by the full-length antibody, e.g., a fragment that retains one or more CDR regions. Examples of antibody 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., sc-Fv; nanobodies; and multispecific antibodies formed from antibody fragments.

[0036] “ Specific binding An antibody to a specific target protein is one that exhibits preferential binding to that target compared to other proteins, but such specificity does not require absolute binding specificity. An antibody is considered to be "specific" for its intended target if binding of the antibody determines the presence of the target protein in a sample, for example, without producing undesirable results such as false positives. The antibodies or binding fragments thereof useful in the present invention will bind to the target protein with an affinity that is at least 2-fold higher, preferably at least 10-fold higher, more preferably at least 20-fold higher, and most preferably at least 100-fold higher than that of a non-target protein. An antibody herein is said to specifically bind to a polypeptide comprising a given amino acid sequence (e.g., the amino acid sequence of a mature human PD-1 molecule) if it binds to a polypeptide comprising that sequence but not to a protein lacking that sequence.

[0037] In this article, the term " Human antibodies ” refers to antibodies that contain only human immunoglobulin protein sequences. Human antibodies may contain murine carbohydrate chains if produced in mice, mouse cells, or hybridomas derived from mouse cells. Similarly, a “mouse antibody” or a “rat antibody” refers to antibodies that contain only mouse or rat immunoglobulin sequences, respectively.

[0038] the term" Humanized antibodies " refers to a form of antibody that contains sequences from non-human (e.g., murine) antibodies as well as human antibodies. Such antibodies contain minimal sequence derived from non-human immunoglobulins. In general, a humanized antibody will comprise essentially all of at least one, and typically two, variable domains, with all or essentially all of its hypervariable loops corresponding to those of a non-human immunoglobulin and all or essentially all of its FRs being those of a human immunoglobulin sequence. The humanized antibody will also optionally comprise 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 the humanized antibody from the parent rodent antibody, the prefix " hum ”, “ hu "or" h" is added to the antibody clone name. A humanized form of a rodent antibody may 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.

[0039] the term" treatment "or" Treating "Treatment" is a method of obtaining beneficial or desired clinical results, including but not limited to one or more of the following: alleviating one or more symptoms caused by the disease, reducing the extent of the disease, stabilizing the disease (e.g., preventing or delaying worsening of the disease), preventing or delaying the spread of the disease (e.g., metastasis), preventing or delaying recurrence of the disease, delaying or slowing the progression of the disease, palliating the disease state, providing remission (partial or complete) of the disease, reducing the dose of one or more other drugs required to treat the disease, delaying the progression of the disease, improving the quality of life, and / or prolonging survival. Therefore, the term "treatment" also includes the reduction of pathological findings of urothelial carcinoma. The methods disclosed herein encompass any one or more of these therapeutic aspects. Anti-PD-1 antibodies

[0040] As disclosed herein, an anti-PD-1 antibody is an antibody or an antigen-binding fragment thereof that specifically binds to human PD-1.

[0041] As disclosed herein, the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody, the heavy chain variable region (V H ) and light chain variable region (V L ) contain complementarity determining regions (CDRs) defined using the Kabat numbering system and listed below:

[0042] As disclosed herein, the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody, the heavy chain variable region (V H ) and light chain variable region (V L ) contains any combination of the CDRs listed below:

[0043] As disclosed herein, the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody, the heavy chain variable region (V H ) and light chain variable region (V L )Include:

[0044] In some embodiments, the antibody comprises an IgG4 Fc region having a serine to proline mutation at position 228 (EU numbering system). In some embodiments, the mutation is referred to as an S228P mutation. In some embodiments, the antibody comprises an IgG4 Fc region having a mutation at one or more of positions 233, 234, 235, 265, 309, and 409 (EU numbering system). For example, in some embodiments, the antibody comprises an IgG4 region having a mutation at 228 and at least one other position, wherein the at least one other mutation results in reduced binding to one or more FcγRs. In further embodiments, the antibody comprises an IgG4 region having a mutation at position 228 and at least 2, at least 3, at least 4, at least 5, or at least 6 additional positions, wherein the one or more additional mutations result in reduced binding to one or more FcγRs. In some embodiments, the antibody comprises an IgG4 region having a mutation at positions 234 and 235. In some embodiments, the antibody comprises an IgG4 region having a mutation at positions 233, 235, and 235. In some embodiments, the antibody comprises an IgG4 region having mutations at positions 234, 235, and 265. In some embodiments, the antibody comprises an IgG4 region having mutations at positions 233, 234, 235, and 265. In some embodiments, the antibody comprises an IgG4 region having mutations at positions 234, 235, 265, and 409. In some embodiments, the antibody comprises an IgG4 region having mutations at positions 233, 234, 235, 265, and 409. In some embodiments, the antibody comprises an IgG4 region having mutations at positions 234, 235, 265, 309, and 409. In some embodiments, the antibody comprises an IgG4 region having mutations at positions 233, 234, 235, 265, 309, and 409. The mutation at position 234 can be a substitution of phenylalanine for valine or a substitution of phenylalanine for alanine. The mutation at position 235 can be a substitution of leucine for alanine. The mutation at position 233 can be a substitution of glutamic acid for proline. The mutation at position 265 can be a substitution of aspartic acid for valine or aspartic acid for threonine. The mutation at position 309 can be a substitution of leucine for valine. The mutation at position 409 can be a substitution of arginine for lysine, threonine, or methionine.

[0045] As disclosed herein, the anti-PD-1 antibody comprises an IgG4 heavy chain constant domain comprising any one of SEQ ID NOs: 83-88 or 91-106.

[0046] As disclosed herein, the anti-PD-1 antibody is an antibody containing F(ab) or F(ab')2, wherein the F(ab) or F(ab')2 comprises the above-mentioned domains (including the heavy chain variable region (V H ), light chain variable region (V L ) and IgG4 heavy chain constant domain).

[0047] As disclosed herein, the anti-PD-1 antibodies comprise a heavy chain variable region (V H ) and light chain variable region (V L ) and an IgG4 heavy chain constant domain comprising SEQ ID NO: 87 or 88, wherein the heavy chain variable region (V H ) and light chain variable region (V L )Include:

[0048] As disclosed herein, the anti-PD-1 antibodies comprise a heavy chain variable region (V H ) and light chain variable region (V L ) and an IgG4 heavy chain effector or constant domain containing SEQ ID NO: 88, wherein the heavy chain variable region (V H ) and light chain variable region (V L ) contain SEQ ID NO:24 and SEQ ID NO:26, respectively.

[0049] As disclosed herein, anti-PD-1 antibodies comprise a uniquely engineered humanized IgG4 Fc domain that reduces FcγRs and thereby reduces antibody-dependent phagocytosis, a potential mechanism of T cell clearance, thereby improving efficacy.

[0050] Anti-PD1 antibodies and antigen-binding fragments thereof can be prepared as described in WO 2015 / 035606 A1, which is incorporated herein by reference. Treatment

[0051] In the treatment methods disclosed herein, a Mab-1 antibody is administered to a patient with UC at a dose. In one embodiment, the patient with UC has not been previously treated with an immunotherapy molecule. In another embodiment, the patient with UC has received anti-cancer systemic therapy (e.g., MVAC). In another embodiment, the patient has previously received platinum adjuvant and neoadjuvant therapy.

[0052] In the treatment method for UC, the anti-PD-1 antibody is administered at a dose of 0.5-10 mg / kg QW or Q2W or Q3W or Q4W. In some embodiments herein, Mab-1 is administered at a dose of 0.5-10 mg / kg QW or Q2W or Q3W. In another treatment regimen, Mab-1 is administered at a dose of 2-10 mg / kg Q2W or Q3W or at a fixed dose of 200 mg Mab-1. Mab-1 can also be administered intravenously at a dose of 2.0 mg / kg Q2W, 5 mg / kg Q2W, 2 mg / kg Q3W or 5 mg / kg Q3W or at a fixed dose of 200 mg of Mab-1.

[0053] In another embodiment, Mab-1 is combined with another agent or agents to treat UC. In one embodiment, Mab-1 is combined with standard therapy for UC (methotrexate, vincristine, doxorubicin, and cisplatin (MVAC)) to treat UC. In yet another embodiment, UC is treated with Mab-1 and cisplatin or cisplatin and gemcitabine. A variation of this treatment is to administer paclitaxel before cisplatin and gemcitabine, and to administer Mab-1 before or during paclitaxel treatment and before or during cisplatin and gemcitabine treatment. Example Study Design—Patient Identification and Recruitment

[0054] The study design aims to enroll patients with UC for dose determination and initial patient identification in Phase 1A and Phase 1B, respectively. Figure 2 In. Figure 2 In the , * indicates the dose expansion regimen; represents a fixed dose that does not exceed the maximum tolerated dose, and Indicates that it is carried out in parallel with Phase 1B.

[0055] Phase 1A was used to determine the safety, RP2D, and preliminary efficacy of Mab-1. In Phase 1A, 10 mg / kg once every two weeks (Q2W) was the maximum dose of Mab-1 administered, and the maximum tolerated dose (MTD) was not reached. In Part 1 of Phase 1A, a dose escalation study was conducted starting from 0.5 mg / kg Q2W to 10 mg / kg Q2W. In Part 2 of Phase 1A, a protocol expansion study was conducted with 2 mg / kg or 5 mg / kg of Mab-1 Q2W or Q3W. In Part 3 of Phase 1A, a fixed-dose expansion study was conducted with 200 mg of Mab-1 Q3W (a fixed dose in selected tumors that did not exceed the exposure at the maximum tolerated dose), and the study was conducted in parallel with Phase 1B. All Phase 1B patients received Mab-1, 5 mg / kg intravenously, Q3W. Radiographic assessment was performed every 8 or 9 weeks according to the Response Evaluation Criteria in Solid Tumors Guidelines Version 1.1 (RECIST v1.1), and an example of this is shown in Figures 6A-6G. Key eligibility criteria for UC subpopulations

[0056] Adult patients (aged ≥18 years) with histologically or cytologically confirmed UC who had at least one measurable lesion (as defined by RECIST v1.1); had received standard therapy but no prior anti-PD-1 or programmed death ligand 1 (PD-L1) therapy; and had an Eastern Cooperative Oncology Group (ECOG) performance status ≤1 were enrolled.

[0057] Patients were excluded if they had a history of severe hypersensitivity reactions to other antibodies. Patients with a history of previous malignancy activity other than UC within the past 2 years and apparently cured localized curable cancers such as basal cell or squamous cell skin cancer, superficial bladder cancer, or carcinoma in situ of the cervix or breast were also excluded.

[0058] Pretreatment tumor specimens were retrospectively assessed for PD-L1 membrane expression by immunohistochemistry on an automated platform. PD-L1 expression status was determined by membrane staining of tumor cells (TC) or tumor-associated immune cells (IC) for PD-L1 of any intensity. PD-L1 was defined as high expression if ≥25% Tc or ≥25% IC expressed membrane PD-L1; and PD-L1 was defined as low / negative if both TC and IC had <25% PD-L1 membrane staining. Eighteen (18) UC patients (n=18) were enrolled in the clinical trial. Patient distribution

[0059] Of the 18 recruited UC patients, 14 had received at least one prior chemotherapy, and 6 had received prior radiotherapy, suggesting the difficulty in treating UC. Figure 3 and Figure 5 As shown, a total of 6 patients (33.3%) continued to receive extended treatment, with at least 3 patients receiving treatment for more than 36 weeks. One of these patients showed a complete response, while the other two showed good partial responses ( Figure 5 Table 1 shows the demographic and disease characteristics of the UC patients. Table 1 Antitumor activity

[0060] Of the 18 UC patients enrolled, 17 were evaluable. Evaluable UC patients were defined as having a measurable baseline tumor assessment and at least one evaluable post-baseline tumor response assessment, or who went on to progress or died before the initial tumor assessment.

[0061] In the clinical trials and results presented below, all patients received Mab-1 at doses of 2.0, 5.0, or 10.0 mg / kg or a fixed dose of 200 mg, administered intravenously every 2 weeks or every 3 weeks.

[0062] Among the 17 evaluable UC patients, one patient achieved a confirmed complete response (CR), four achieved a confirmed partial response (PR), and three achieved stable disease (SD). The disease control rate (DCR = CR + PR + SD) was found to be 47.1% (n = 8 / 17). The median duration of treatment was 3.0 months, the longest duration was 18.3 months, and the shortest duration was 0.7 months. The anti-tumor activity of Mab-1 was shown in Figures 3 to 5 middle. PD-L1 status response

[0063] PD-L1 status and clinical response in UC patients treated with Mab-1 at doses of 2.0, 5.0, or 10.0 mg / kg or a fixed dose of 200 mg intravenously Q2W or Q3W were also evaluated.

[0064] A total of 10 patients had evaluable PD-L1 status and clinical response. Clinical responses were observed in patients with both high and low PD-L1 expression (Tables 2 and Figure 4 Table 2 shows the best overall response for each evaluable patient, as confirmed by PD-L1 status. Table 2: Best overall response per evaluable patient confirmed by PD-L1 status *PD-L1 status could not be assessed due to insufficient tumor tissue. PD-L1 high was defined as ≥25% of tumor cells (TC) or ≥25% of tumor-associated immune cells (IC); if both TC and IC had <25% PD-L1 staining, PD-L1 low / negative.

[0065] The study found that PD-L1 - disease compared to PD-L1 + The objective response rate in patients with PD-L1 disease was higher, indicating that Mab-1 + UC patients can be effective. Safety and tolerability

[0066] This clinical study also found that Mab-1 treatment was generally well tolerated in UC patients. Fifteen of the 18 UC patients experienced treatment-related adverse events (TRAEs) (Table 3), of which fatigue (n=1), hyperglycemia (n=1), and type 1 diabetes (n=1) were the only treatment-related adverse events with a severity of ≥ grade 3 (regardless of attribution).

[0067] One patient discontinued treatment due to an infusion-related reaction related to Mab-1, and another patient had a treatment-emergent adverse event with fatal outcome (muscle weakness) that was unrelated to Mab-1 treatment. This data is shown in Table 3. Table 3: Treatment-related adverse events occurring in ≥2 patients with UC

[0068] These results confirm that the antibody of the present application (Mab-1) is tolerable and its toxicity profile demonstrates that adverse events (AEs) are generally of low severity, manageable, and reversible.

[0069] The foregoing examples and descriptions of certain embodiments should be considered illustrative, rather than limiting, of the present invention as defined by the claims. As will be readily appreciated, many variations and combinations of the features described above may be employed without departing from the present invention as set forth in the claims. All such variations are intended to be within the scope of the present invention. All references cited are incorporated herein by reference in their entirety.

[0070] It will be appreciated that, even if a prior art publication is referred to herein, this reference does not constitute an admission that the publication forms part of the common general knowledge in the art in any country.

[0071] The present disclosure relates to the following embodiments. 1. A method for treating a patient with urothelial carcinoma, the method comprising administering to the patient a therapeutically effective amount of an anti-PD-1 antibody or an antigen-binding fragment thereof that reduces FcγR binding and reduces antibody-dependent phagocytosis. 2. The method according to embodiment 1, wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ), the heavy chain variable region (V H ) and light chain variable region (V L ) contains the following complementarity determining regions (CDRs): a) mu317 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NOs: 11, 12, 13); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 14, 15, 16); b) mu326 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NO: 17, 18, 19); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 20, 21, 22); c) 317-4B6 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NOs: 31, 32, 33); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 34, 35, 36); d) 326-4A3 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NO: 37, 38, 39); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 40, 41, 42); e) 317-1H CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NOs: 11, 59, 13); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 14, 15, 16); f) 317-4B2 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NOs: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 317-4B2 CDR-H1, 60, 13); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 61, 15, 16); g) 317-4B5 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NO: 11, 60, 13); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 61, 15, 16); h) 317-4B6 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NOs: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 317-4B6 CDR-H1, 32, 13); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 61, 15, 16); i) 326-1 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NO: 17, 62, 19); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 20, 21, 22); j) 326-3B1 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NO: 17, 62, 19); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 20, 21, 22); or k) 326-3G1 CDR-H1, CDR-H2 and CDR-H3 (SEQ ID NO: 17, 62, 19); and CDR-L1, CDR-L2 and CDR-L3 (SEQ ID NO: 20, 21, 22). 3. The method according to embodiment 1, wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody, the heavy chain variable region (V H ) and light chain variable region (V L ) contains the following CDRs: (a) CDR-H1 (SEQ ID NO 31), CDR-H2 (SEQ ID NO 32) and CDR-H3 (SEQ ID NO 33), CDR-L1 (SEQ ID NO 34), CDR-L2 (SEQ ID NO 35) and CDR-L3 (SEQ ID NO 36); or (b) CDR-H1 (SEQ ID NO 37), CDR-H2 (SEQ ID NO 38) and CDR-H3 (SEQ ID NO 39), CDR-L1 (SEQ ID NO 40), CDR-L2 (SEQ ID NO 41) and CDR-L3 (SEQ ID NO 42). 4. The method according to embodiment 1, wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody, the heavy chain variable region (V H ) and light chain variable region (V L )Include: 5. The method of any one of embodiments 2-4, wherein the anti-PD-1 antibody comprises an IgG4 heavy chain constant domain as shown in any one of SEQ ID NOs: 83-88 or 91-106. 6. The method of any one of embodiments 2-3, wherein the anti-PD-1 antibody comprises a F(ab) or F(ab')2 comprising the CDRs of embodiment 2 or 3, or the heavy chain variable region (V H ) and light chain variable region (V L ). 7. The method according to embodiment 1, wherein the anti-PD-1 antibody comprises an IgG4 heavy chain constant domain as shown in SEQ ID NO: 87 or 88, and wherein the heavy chain variable region (V H) and light chain variable region (V L )Include: a) mu317 (SEQ ID NOs: 4 and 6, respectively); p) 317-3H1 (SEQ ID NOs: 69 and 26, respectively); b) mu326 (SEQ ID NOs: 8 and 10, respectively); q) 317-311 (SEQ ID NOs: 70 and 26, respectively); c) 317-4B6 (SEQ ID NOs: 24 and 26, respectively); d) 326-4A3 (SEQ ID NOs: 28 and 30, respectively); r) 317-4B1 (SEQ ID NOs: 71 and 26, respectively); e) 317-4B2 (SEQ ID NOs: 43 and 44, respectively); s) 317-4B3 (SEQ ID NOs: 72 and 26, respectively); f) 317-4B5 (SEQ ID NOs: 45 and 46, respectively); t) 317-4B4 (SEQ ID NOs: 73 and 26, respectively); g) 317-1 (SEQ ID NOs: 48 and 50, respectively); u) 317-4A2 (SEQ ID NOs: 74 and 26, respectively); h) 326-3B1 (SEQ ID NOs: 51 and 52, respectively); v) 326-3A1 (SEQ ID NOs: 75 and 30, respectively); i) 326-3GI (SEQ ID NOs: 53 and 54, respectively); w) 326-3C1 (SEQ ID NOs: 76 and 30, respectively); j) 326-1 (SEQ ID NOs: 56 and 58, respectively); x) 326-3D1 (SEQ ID NOs: 77 and 30, respectively); k) 317-3A1 (SEQ ID NOs: 64 and 26, respectively); y) 326-3E1 (SEQ ID NOs: 78 and 30, respectively); l) 317-3C1 (SEQ ID NOs: 65 and 26, respectively); z) 326-3F1 (SEQ ID NOs: 79 and 30, respectively); m) 317-3E1 (SEQ ID NOs: 66 and 26, respectively); aa) 326-3B N55D (SEQ ID NOs: 80 and 30, respectively); n) 317-3F1 (SEQ ID NOs: 67 and 26, respectively); ab) 326-4A1 (SEQ ID NOs: 28 and 81, respectively); or o) 317-3G1 (SEQ ID NOs: 68 and 26, respectively); ac) 326-4A2 (SEQ ID NOs: 28 and 82, respectively). 8. The method of embodiment 1, wherein the anti-PD-1 antibody comprises an IgG4 heavy chain domain comprising SEQ ID NO: 88, and wherein the heavy chain variable region (V H ) and light chain variable region (V L ) are shown as SEQ ID NO: 24 and SEQ ID NO: 26, respectively. 9. The method of embodiment 1, wherein the anti-PD-1 antibody comprises an IgG4 Fc region comprising an S228P mutation at position 228 and amino acid mutations at positions 233, 234, and 235, wherein the mutations at positions 233, 234, and 235 reduce binding to at least one Fcγ receptor relative to Fcγ binding of a reference IgG4 antibody having only the mutation at position 228. 10. The method of embodiment 9, wherein the IgG4 Fc region comprises amino acid mutations at positions 228, 233, 234, 235, and 265. 11. The method of embodiment 9, wherein the IgG4 Fc region comprises amino acid mutations at positions 228, 233, 234, 235, 265, 309, and 409. 12. The method of embodiment 9, wherein the IgG4 Fc region consists of amino acid mutations of S228P, E233P, F234V, and L235A. 13. The method of embodiment 1, wherein the urothelial cancer is a cancer of the ureter, urethra, renal pelvis and / or bladder. 14. The method of embodiment 13, wherein the urothelial carcinoma is transitional cell carcinoma. 15. The method of embodiment 13, wherein the urothelial carcinoma is advanced or metastatic. 16. The method according to embodiment 13, wherein the urothelial cancer is PD-L1 + Urothelial carcinoma or PD-L1 - Urothelial carcinoma. 17. The method according to embodiment 13, wherein the urothelial cancer is PD-L1+ Urothelial carcinoma. 18. The method of embodiment 1, wherein the anti-PD-1 antibody is administered parenterally at a dose of 0.5-10 mg / kg QW, or Q2W, or Q3W, or Q4W. 19. The method of embodiment 18, wherein the anti-PD-1 antibody is administered at a dose of 2-10 mg / kg QW, Q2W, or Q3W. 20. The method of embodiment 18, wherein the anti-PD-1 antibody is administered at a dose of 2-10 mg / kg or at a fixed dose of 200 mg Q2W or Q3W. 21. The method of embodiment 18, wherein the anti-PD-1 antibody is administered intravenously at a dose of 2.0 mg / kg Q2W, 5 mg / kg Q2W, 2 mg / kg Q3W, or 5 mg / kg Q3W, or at a fixed dose of 200 mg Q2W or Q3W. 22. The method of embodiment 1, further comprising administering an additional therapeutic agent. 23. The method of embodiment 22, wherein the additional therapeutic agent is methotrexate, vincristine, doxorubicin, and cisplatin (MVAC). 24. The method of embodiment 22, wherein the additional therapeutic agent is cisplatin. 25. The method of embodiment 24, wherein cisplatin is administered together with gemcitabine. 26. The method of embodiment 22, wherein the additional therapeutic agent is paclitaxel. 27. The method of embodiment 25, wherein paclitaxel is administered before cisplatin and gemcitabine.

Claims

1. A method for treating a patient with urothelial carcinoma, the method comprising administering to the patient a therapeutically effective amount of an anti-PD-1 antibody or an antigen-binding fragment thereof that reduces FcγR binding and reduces antibody-dependent phagocytosis.

2. The method of claim 1, wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ), the heavy chain variable region (V H ) and light chain variable region (V L ) contains the following complementarity determining regions (CDRs):

3. The method of claim 1, wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody, the heavy chain variable region (V H ) and light chain variable region (V L ) contains the following CDRs:

4. The method of claim 1, wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody, the heavy chain variable region (V H ) and light chain variable region (V L )Include:

5. The method of any one of claims 2-4, wherein the anti-PD-1 antibody comprises an IgG4 heavy chain constant domain as shown in any one of SEQ ID NOs: 83-88 or 91-106.

6. The method of any one of claims 2-3, wherein the anti-PD-1 antibody comprises a F(ab) or F(ab')2 comprising the CDRs of claim 2 or 3, or the heavy chain variable region (V H ) and light chain variable region (V L ).

7. The method of claim 1, wherein the anti-PD-1 antibody comprises an IgG4 heavy chain constant domain as shown in SEQ ID NO: 87 or 88, and wherein the heavy chain variable region (V H ) and light chain variable region (V L )Include:

8. The method of claim 1, wherein the anti-PD-1 antibody comprises an IgG4 heavy chain domain comprising SEQ ID NO: 88, and wherein the heavy chain variable region (V H ) and light chain variable region (V L ) are shown as SEQ ID NO:24 and SEQ ID NO:26, respectively.

9. The method of claim 1 , wherein the anti-PD-1 antibody comprises an IgG4 Fc region comprising an S228P mutation at position 228 and amino acid mutations at positions 233, 234, and 235, wherein the mutations at positions 233, 234, and 235 reduce binding to at least one Fcγ receptor relative to Fcγ binding of a reference IgG4 antibody having only the mutation at position 228.

10. The method of claim 9, wherein the IgG4 Fc region comprises amino acid mutations at positions 228, 233, 234, 235, and 265.

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