Method for treating solid tumor

Abstract

Use of a CD73 antibody or antigen-binding fragment thereof for the preparation of a medicament for the treatment of a solid tumor in a subject in need thereof, wherein the medicament further comprises a PD-1 antagonist.

Description

Technical Field

[0001] The present application relates to a method for treating solid tumors. In particular, a method for treating solid tumors (e.g., advanced solid tumors) by using an anti-CD73 antibody or an antigen-binding fragment thereof is provided. Background Art

[0002] CD73, cluster of differentiation 73, also known as 5'-nucleotidase (5'-NT) or ecto-5'-nucleotidase, is an enzyme used to convert AMP to adenosine. CD73 catalyzes the formation of extracellular adenosine, which contributes to the immunosuppressive tumor environment. CD73 is overexpressed in stromal cells and multiple types of tumor cells, as well as Treg, M2 and myeloid-derived suppressor cells (MDSC).

[0003] Preclinical evidence shows that CD73 inhibition prevents adenosine-mediated lymphocyte suppression, enhances the activity of CD8+ effector cells, and reduces MDSCs and Tregs. Several anti-CD73 antibodies are being developed as potential anticancer drugs, but none has been approved for clinical use. Summary of the invention

[0004] This application meets clinical treatment needs by using CD73 antibodies.

[0005] In one aspect, the present application provides a method for treating solid tumors, comprising administering a CD73 antibody or an antigen-binding fragment thereof and a PD-1 antagonist to a subject in need thereof at a dose of about 2 to about 60 mg / kg body weight.

[0006] In some embodiments of the methods of the present application, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 15 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 10 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 10 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 5 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 20 to about 30 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2, about 5, about 10, about 15, or about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 20 or 30 mg / kg body weight.

[0007] In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject once a week (QW), once every two weeks (Q2W), once every three weeks (Q3W), once every four weeks (Q4W), or once a month. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject once every three weeks (Q3W). In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of 20 or 30 mg / kg, once every three weeks (Q3W). In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject intravenously.

[0008] In some embodiments, the CD73 antibody or its antigen-binding fragment is selected from the group consisting of a full-length antibody, Fab, Fab', F(ab')2, scFv and sdAb. In some embodiments, the CD73 antibody or its antigen-binding fragment is a full-length antibody. In some embodiments, the CD73 antibody or its antigen-binding fragment comprises HCDR1, HCDR2 and HCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the heavy chain variable region (VH) as shown in SEQ ID NO: 1, wherein CDR1, CDR2 and CDR3 are according to the Kabat numbering scheme. In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises: (1) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO:3 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:3; (2) HCDR2 comprising the amino acid sequence as shown in SEQ ID NO:4 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:4; and (3) HCDR3 comprising the amino acid sequence as shown in SEQ ID NO:5 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:5. In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising the amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 90% identity to SEQ ID NO:1.

[0009] In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises LCDR1, LCDR2, and LCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2, and CDR3 in the light chain variable region (VL) as shown in SEQ ID NO:2, wherein CDR1, CDR2, and CDR3 are according to the Kabat numbering scheme. In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises: (1) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO:6 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:6; (2) LCDR2 comprising the amino acid sequence as shown in SEQ ID NO:7 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:7; and (3) LCDR3 comprising the amino acid sequence as shown in SEQ ID NO:8 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:8. In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises a light variable region (VL) comprising an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 90% identical to SEQ ID NO:2.

[0010] In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises: (1) HCDR1, HCDR2 and HCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the heavy chain variable region (VH) as shown in SEQ ID NO:1, and (2) LCDR1, LCR2 and LCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the light chain variable region (VL) as shown in SEQ ID NO:2, wherein CDR1, CDR2 and CDR3 are according to the Kabat numbering scheme. In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises: (1) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO:3, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:3; (2) HCDR2 comprising the amino acid sequence as shown in SEQ ID NO:4, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:4; (3) HCDR3 comprising the amino acid sequence as shown in SEQ ID NO:5, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:5; (4) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO:6, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:6; (5) LCDR2 comprising the amino acid sequence as shown in SEQ ID NO:7, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:7; and (6) LCDR3 comprising the amino acid sequence as shown in SEQ ID NO:8, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:8.

[0011] In some embodiments, the CD73 antibody or antigen-binding fragment thereof comprises: (1) a heavy chain variable region (VH) comprising an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence that is at least 90% identical to SEQ ID NO:1, and (2) a light chain variable region (VL) comprising an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 90% identical to SEQ ID NO:2.

[0012] In some embodiments, the CD73 antibody or antigen-binding fragment thereof is selected from the group consisting of uliledlimab (I-Mab Biopharma), oleclumab (AstraZeneca), CPI-006 (Corvus Pharma), BMS-986179 (Bristol-Myers Squibb), AB-680 (Arcus Biosciences), NZV-930 (SRF373, Surface Oncolohgy / Novartis), JAB-BX102 (Jacobio), AK119 (Akesobio), Sym024 (Symphogen), IBI 325 (Innovent), BR 101 (Hisun BioRay), and LY3475070 (Eli Lilly).

[0013] In some embodiments, the solid tumor is selected from the group consisting of ovarian cancer, sarcoma, cervical cancer, bladder cancer, breast cancer, colorectal cancer, endometrial cancer, esophageal cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, lymphoma, acute lymphoblastic leukemia, multiple myeloma, glioma, mesothelioma, melanoma, skin cancer (basal cell carcinoma), pancreatic cancer, prostate cancer, uterine cancer and thyroid cancer.

[0014] In some embodiments, lung cancer includes non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). In some embodiments, the solid tumor is an advanced solid tumor. In some embodiments, the solid tumor is a metastatic solid tumor. In some embodiments, the solid tumor is a recurrent solid tumor. In certain embodiments, the solid tumor is a refractory solid tumor. In some embodiments, the solid tumor is in phase III or phase IV.

[0015] In some embodiments, the CD73 antibody or antigen-binding fragment thereof and the PD-1 antagonist are administered simultaneously. In some embodiments, the CD73 antibody or antigen-binding fragment thereof and the PD-1 antagonist are administered separately.

[0016] In some embodiments, the PD-1 antagonist is a PD-1 antibody. In some embodiments, the PD-1 antibody comprises: (1) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 10 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 10; (2) LCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 11 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 11; (3) LCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 12 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 12; (4) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 13 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 13; (5) HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 14 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 14; and (6) HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 15 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 15. In some embodiments, the PD-1 antibody comprises: (1) a heavy chain variable region (VH) comprising an amino acid sequence as shown in SEQ ID NO: 16 or an amino acid sequence having at least 90% identity with SEQ ID NO: 16; and (2) a light chain variable region (VL) comprising an amino acid sequence as shown in SEQ ID NO: 17 or an amino acid sequence having at least 90% identity with SEQ ID NO: 17.

[0017] In some embodiments, the PD-1 antibody is selected from the group consisting of: pembrolizumab, nivolumab, toripalimab, pidilizumab, cemiplimab, sintilimab, cetrelimab, spartalizumab, camrelizumab, tislelizumab, balstilimab, dostarlimab, ABBV-181, penpulimab, genolimzumab ), retifanlimab, sasanlimab, AMP-224, AB122, F-520, MEDI-3387, MEDI-5771, MEDI-0680, SG-001, BCD-100, BAT-1306, BI-754091, CBT-501, GLS-010, LZM-009, Sym-021, CS-1003, HLX-10, AK-103, AM-0001, ENUM-244C8, ENUM-388D4, JTX-4014, RXI-762, STI-A1110, HLX-20, SSI-361, APL-501, TJ0141H, and SNA-01. In some embodiments, the PD-1 antibody is toripalimab. In some embodiments, the PD-1 antibody is administered to the subject at a fixed dose of 240 mg once every three weeks (Q3W).

[0018] In some embodiments, the subject has tumor tissue in which at least 40% of the tumor cells are CD73 positive. In some embodiments, the subject has tumor tissue in which at least 30%, 35%, 40%, 45%, or 50% of the tumor cells are CD73 positive. In some embodiments, at least 1% of the tumor cells are PD-L1 positive.

[0019] In another aspect, the present application provides a use of a CD73 antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating a solid tumor in a subject in need thereof, wherein the medicament is administered to the subject in combination with a PD-1 antagonist at a dose of about 2 to about 60 mg / kg body weight.

[0020] In another aspect, the present application provides a product comprising (1) a CD73 antibody or an antigen-binding fragment thereof, and (2) a package insert suggesting that the CD73 antibody or the antigen-binding fragment thereof be administered in combination with a PD-1 antagonist to a subject in need thereof at a dose of about 2 to about 30 mg / kg body weight. In some embodiments, the product further comprises a PD-1 antagonist.

[0021] It should be understood that one, some or all of the various embodiments described herein may be combined to form other embodiments of the present invention. For those skilled in the art, these and other aspects of the present invention are apparent. The following detailed description of the invention will further set forth these and other embodiments of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Shown is the simulated pharmacokinetic profile of uliledlimab at the proposed regimen of 30 mg / kg, Q3W (with additional dose on C1D8) using a population pharmacokinetic model developed based on observed pharmacokinetic results.

[0023] Figures 2A-2F Shown are the expression levels of total and free CD73 as determined by immunohistochemistry (IHC) staining and the residual CD73 enzyme activity as determined by enzyme histochemistry (EHC) staining in tumor biopsies obtained from two NSCLC patients treated with 30 mg / kg Uliledlimab. 波谷 Tumor biopsies were collected on Cycle 4 Day 1 (C4D1).

[0024] Figure 3 Receiver operating characteristic (ROC) analysis to identify cut-off values ​​for CD73 levels is shown.

[0025] Figure 4 It was shown that in patients with higher CD73 expression (≥40%), especially in those with PD-L1 TPS1-49% and ≥50%, the combination therapy achieved better clinical responses. DETAILED DESCRIPTION definition

[0026] Before describing the embodiments in detail, it should be understood that the present disclosure is not limited to specific compositions or biological systems, which, of course, can vary. It should also be understood that the terms used in the present application are only for the purpose of describing specific embodiments and are not intended to be restrictive.

[0027] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural references unless the content clearly dictates otherwise. Thus, for example, reference to "a molecule" may optionally include a combination of two or more such molecules, and so forth.

[0028] The term "about" used in this application refers to the usual error range of each numerical value known to those skilled in the art. "About" a value or parameter mentioned in this application includes (and describes) embodiments for the value or parameter itself.

[0029] It should be understood that various aspects and embodiments of the present disclosure include "comprising / including" various aspects and embodiments, "consisting of various aspects and embodiments", and "consisting essentially of various aspects and embodiments.

[0030] As used herein, the term "antibody" is used in the broadest sense and specifically covers intact antibodies (such as full-length antibodies), antibody fragments (including but not limited to Fab, F(ab')2, scFv, scFv-Fc, single domain antibodies, single heavy chain antibodies and single light chain antibodies), monoclonal antibodies and polyclonal antibodies, so long as they exhibit the desired biological activity (such as epitope binding).

[0031] As used herein, the term "isolated" antibody may refer to an antibody that is substantially free of other cellular material. In one embodiment, an isolated antibody is substantially free of other proteins from the same species. In another embodiment, an isolated antibody is expressed by cells from a different species and is substantially free of other proteins from the different species. In some embodiments, an "isolated" antibody refers to an antibody that has been identified and separated and / or recovered from components of its natural environment, wherein contaminating components of its natural environment refer to substances that interfere with the diagnostic or therapeutic use of the antibody and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. The antibody may be isolated using protein purification techniques well known in the art such that the antibody is substantially free of naturally associated components (or components associated with the cellular expression system used to produce the antibody). In some embodiments, the antibody will be purified: (1) greater than 75% by weight of the antibody as determined by the Lowry method, and most preferably greater than 80%, 90%, 95%, or 99% by weight; or (2) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or, preferably, silver stain. Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step.

[0032] As used in this application, the terms "natural antibodies and immunoglobulins" are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light chains (L) and two identical heavy chains (H). Each light chain is connected to the heavy chain by one covalent disulfide bond (also called a "VH / VL pair"), while the number of disulfide bonds between the heavy chains of different immunoglobulin isotypes varies. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has a variable domain (VH) at one end, followed by a number of constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the variable domain of the light chain is aligned with the variable domain of the heavy chain. It is believed that specific amino acid residues form an interface between the light chain and heavy chain variable domains. See, for example, Chothia et al., J. Mol. Biol., 186:651 (1985); Novotny and Haber, Proc. Natl. Acad. Sci. USA, 82:4592 (1985).

[0033] As used herein, the term "variable" refers to the fact that some parts of the variable domains have great differences in sequence between antibodies and are used for the binding and specificity of each specific antibody to its specific antigen. However, variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called complementary determining regions (CDRs) or hypervariable regions in both the light chain and heavy chain variable domains. The more conservative part of the variable domain is called framework (FR). The variable domains of natural heavy and light chains each contain four FR regions, most of which adopt a β sheet structure, connected by three CDRs, which form a loop connecting the β sheet structure, and in some cases constitute a part of the β sheet structure. The CDRs on each chain are kept together in close proximity by the FR region, and together with the CDRs on the other chain, contribute to the formation of the antigen binding site of the antibody. For example, see Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Maryland, (1991). The constant domains are not directly involved in the binding of an antibody to an antigen, but exhibit various effector functions, such as participation in antibody-dependent cellular cytotoxicity. Variable region sequences of interest include humanized variable region sequences of CD47 antibodies described in detail elsewhere in this application.

[0034] The term "hypervariable region (HVR)" or "complementarity determining region (CDR)" may refer to a subdomain of a VH and VL domain characterized by enhanced sequence variability and / or forming defined loops. These include three CDRs (H1, H2, and H3) in the VH domain and three CDRs (L1, L2, and L3) in the VL domain. H3 is considered to be key to conferring good binding specificity, while L3 and H3 show the highest levels of diversity. See Johnson and Wu, in Methods in Molecular Biology 248: 1-25 (Lo ed., Human Press, Totowa, NJ, 2003).

[0035] Many descriptions of CDR / HVR are known. Kabat complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used complementarity determining regions (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, Maryland (1991)). Chothia refers to the position of structural loops (Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987)). AbM HVR represents a compromise between Kabat HVR and Chothia structural loops and is adopted by Oxford Molecular's AbM antibody modeling software. "Contact" HVRs are based on analysis of existing complex crystal structures. The residues of each of these HVR / CDRs are recorded as follows: "framework" or "FR" residues refer to those variable domain residues other than HVR / CDR residues.

[0036] Also known are "extended" HVRs: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL, 26-35 (H1), 50-65 or 49-65 (H2), and 93-102, 94-102 or 95-102 (H3) in VH (Kabat numbering).

[0037] "Numbering according to Kabat" may refer to the numbering system used for the heavy chain variable domain or light chain variable domain compiled by Kabat et al. above. The actual linear amino acid sequence may contain fewer or additional amino acids, corresponding to the shortening of the FR or HVR of the variable domain or the insertion of the FR or HVR of the variable domain. The Kabat numbering of the residues of a given antibody can be determined by comparing the homologous region of the antibody sequence with the "standard" Kabat numbering sequence. Typically, when referring to the residues in the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain), Kabat numbering is used, and when referring to the residues in the heavy chain constant region, the EU numbering system or index (such as the EU index in Kabat, according to the numbering of EU IgG1) is generally used.

[0038] As used herein, the term "antibody fragment" and all grammatical variants thereof are defined as a portion of an intact antibody that contains the antigen binding site or variable region of the intact antibody and, in some cases, does not contain the constant heavy chain domains (i.e., CH2, CH3, and / or CH4, depending on the antibody isotype) of the Fc region of the intact antibody. Examples of antibody fragments include Fab, Fab', Fab'-SH, F(ab')2, and Fv fragments; diabodies; any antibody fragment of a polypeptide having a primary structure consisting of an uninterrupted sequence of continuous amino acid residues (referred to herein as a "single-chain antibody fragment" or "single-chain polypeptide"), including but not limited to (1) single-chain Fv (scFv) molecules, (2) a single-chain polypeptide containing only one light chain variable domain or a fragment thereof containing the three CDRs of the light chain variable domain without the associated heavy chain portion, and (3) a single-chain polypeptide containing only one heavy chain variable region or a fragment thereof containing the three CDRs of the heavy chain variable region without the associated light chain portion; and multispecific or multivalent structures formed by antibody fragments. In antibody fragments comprising one or more heavy chains, the heavy chain may comprise any constant domain sequence found in the non-Fc region of an intact antibody (such as CH1 in IgG isotypes), and / or may comprise any hinge region sequence found in an intact antibody, and / or may comprise a leucine zipper sequence fused to or located within a hinge region sequence or a constant domain sequence of a heavy chain.

[0039] The Fab fragment also contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. The Fab' fragment differs from the Fab fragment by the addition of several residues to the carboxyl terminus of the heavy chain CH1 domain, including one or more cysteines in the antibody hinge region. Fab'-SH herein refers to Fab' in which the cysteine ​​residues of the constant domains carry free thiol groups. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0040] As used herein, the term "monoclonal antibody" (mAb) refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies constituting the population are identical except for a small amount of possible naturally occurring mutations. Monoclonal antibodies are highly specific and are directed only to a single antigenic site. Each mAb is directed to a single determinant on the antigen. In addition to its specificity, the advantage of monoclonal antibodies is that they can be synthesized by hybridoma culture without being contaminated by other immunoglobulins. The modifier "monoclonal" indicates that the characteristic of the antibody is obtained from a substantially homogeneous antibody population, and should not be construed as requiring the production of antibodies by any particular method. For example, the monoclonal antibodies used according to the disclosure of the present application can be made in immortalized B cells or their hybridomas, or can also be made by DNA recombination methods.

[0041] The monoclonal antibodies in the present application include hybrid antibodies and recombinant antibodies produced by splicing the variable (including hypervariable) domains of CD47 antibodies with constant domains (e.g., "humanized" antibodies), or splicing light chains with heavy chains, or splicing chains from one species with chains from another species, or fusion with heterologous proteins, regardless of their species of origin or immunoglobulin class or subclass designations, as well as antibody fragments (e.g., Fab, F(ab')2, and Fv), as long as they exhibit the desired biological activity.

[0042] The monoclonal antibodies in the present application specifically include chimeric antibodies (immunoglobulins) in which a portion of the heavy chain and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a specific species or belonging to a specific antibody class or subclass, while the remainder of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, as long as they exhibit the desired biological activity.

[0043] As used in this application, the term "treatment" refers to a clinical intervention intended to change the natural course of the treated individual or cell in the clinical pathological process. The ideal therapeutic effect includes reducing the rate of disease progression, improving or alleviating the disease state, and alleviating or improving the prognosis. For example, if one or more symptoms related to cancer are alleviated or eliminated, including but not limited to reducing the proliferation of cancer cells (or eliminating cancer cells), reducing the symptoms caused by the disease, improving the quality of life of individuals with the disease, reducing the dosage of other drugs required for treating the disease, and / or extending the survival period of the individual, then the individual has successfully obtained "treatment". In some embodiments, "treating" a disease (such as cancer) refers to delaying the development of the disease, that is, postponing, hindering, slowing down, delaying, stabilizing and / or delaying the development of a disease (such as cancer). This delay time varies in length, depending on the disease and / or the medical history of the individual being treated. As is apparent to those skilled in the art, sufficient or significant delays can actually include prevention, which is that the individual will not have disease development, for example, advanced cancer (such as metastasis) may be postponed.

[0044] As used herein, the term "subject" for purposes of treatment refers to any animal classified as a mammal, including humans, livestock and farm animals, and zoo, sport or pet animals, such as dogs.

[0045] As used herein, the term "advanced solid tumor" refers to a solid tumor that cannot be cured or grows beyond the original site of origin, whether locally advanced or metastatic. In some embodiments, advanced solid tumors include but are not limited to solid tumors in stage III or stage IV.

[0046] As used herein, the term "metastatic" or "metastasis" refers to the spread of a tumor from its initial or primary site to different or secondary sites in a subject's body. It is generally distinguished from cancer invasion, which refers to the direct spread and infiltration of cancer cells into adjacent tissues.

[0047] As used herein, the term "refractory" refers to a disorder, disease or condition that has not responded to prior treatment, which may include one or more lines of therapy. CD73 Antibody

[0048] CD73, cluster of differentiation 73, also known as 5'-nucleotidase (5'-NT) or ecto-5'-nucleotidase, is an enzyme used to convert AMP to adenosine. CD73 catalyzes the formation of extracellular adenosine, which contributes to the immunosuppressive tumor environment. CD73 is overexpressed in stromal cells and multiple types of tumor cells, as well as Treg, M2 and myeloid-derived suppressor cells (MDSC).

[0049] Any CD73 antibody known in the art can be used in the present application. In some embodiments, the CD73 antibody is selected from the group consisting of a full-length antibody, Fab, Fab', F(ab')2, scFv, and sdAb. In some embodiments, the CD73 antibody is a full-length antibody.

[0050] In some embodiments, the CD73 antibody comprises HCDR1, HCDR2, and HCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2, and CDR3 in the heavy chain variable region (VH) as shown in SEQ ID NO: 1, wherein CDR1, CDR2, and CDR3 are according to the Kabat numbering scheme. In some embodiments, the CD73 antibody comprises: (1) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 3 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 3; (2) HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 4 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 4; and (3) HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 5 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 5.

[0051] In some embodiments, the CD73 antibody comprises a heavy chain variable region (VH) comprising an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence that is at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:1.

[0052] In some embodiments, the CD73 antibody comprises LCDR1, LCR2, and LCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2, and CDR3 in the light chain variable region (VL) shown in SEQ ID NO:2, wherein CDR1, CDR2, and CDR3 are according to the Kabat numbering scheme. In some embodiments, the CD73 antibody comprises: (1) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO:6 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:6; (2) LCDR2 comprising the amino acid sequence as shown in SEQ ID NO:7 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:7; and (3) LCDR3 comprising the amino acid sequence as shown in SEQ ID NO:8 or an amino acid sequence having one or more substitutions compared to SEQ ID NO:8.

[0053] In some embodiments, the CD73 antibody comprises a light chain variable region (VL) comprising an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:2.

[0054] In some embodiments, the CD73 antibody comprises: (1) HCDR1, HCDR2 and HCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the heavy chain variable region (VH) as shown in SEQ ID NO:1, and (2) LCDR1, LCR2 and LCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the light chain variable region (VL) as shown in SEQ ID NO:2, wherein CDR1, CDR2 and CDR3 are according to the Kabat numbering scheme.

[0055] In some embodiments, the CD73 antibody comprises: (1) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO:3, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:3; (2) HCDR2 comprising the amino acid sequence as shown in SEQ ID NO:4, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:4; (3) HCDR3 comprising the amino acid sequence as shown in SEQ ID NO:5, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:5; (4) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO:6, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:6; (5) LCDR2 comprising the amino acid sequence as shown in SEQ ID NO:7, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:7; and (6) LCDR3 comprising the amino acid sequence as shown in SEQ ID NO:8, or an amino acid sequence having one or more substitutions compared to SEQ ID NO:8.

[0056] In some embodiments, the CD73 antibody comprises: (1) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 1, or an amino acid sequence at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1, and (2) a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:2, or an amino acid sequence at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:2.

[0057] In some embodiments, the CD73 antibody is selected from the group consisting of uliledlimab (I-Mab Biopharma), oleclumab (AstraZeneca), CPI-006 (Corvus Pharma), BMS-986179 (Bristol-Myers Squibb), AB-680 (Arcus Biosciences), NZV-930 (SRF373, Surface Oncolohgy / Novartis), JAB-BX102 (Jacobio), AK119 (Akesobio), Sym024 (Symphogen), IBI325 (Innovent), BR101 (Hisun BioRay), and LY3475070 (Eli Lilly). In some embodiments, the CD73 antibody is uliledlimab (I-Mab Biopharma) or oleclumab (AstraZeneca). PD-1 antagonists

[0058] Any suitable PD-1 antagonist known in the art may be used in the present application. In some embodiments, the PD-1 antagonist includes a small molecule inhibitor of PD-1 signaling. In some embodiments, the PD-1 antagonist includes an antibody or an antigen-binding fragment thereof, such as a PD-1 antibody or an antigen-binding fragment thereof. In some embodiments, the PD-1 antagonist is a PD-1 antibody or an antigen-binding fragment thereof.

[0059] In some embodiments, the PD-1 antibody comprises HCDR1, HCDR2, and HCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2, and CDR3 in the heavy chain variable region (VH) as shown in SEQ ID NO: 17, wherein CDR1, CDR2, and CDR3 are according to the Kabat numbering scheme. In some embodiments, the CD73 antibody comprises: (1) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 13 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 13; (2) HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 14 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 14; and (3) HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 15 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 15.

[0060] In some embodiments, the PD-1 antibody comprises a heavy chain variable region (VH) comprising an amino acid sequence as shown in SEQ ID NO: 17, or an amino acid sequence having at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17.

[0061] In some embodiments, the PD-1 antibody comprises LCDR1, LCR2, and LCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2, and CDR3 in the light chain variable region (VL) shown in SEQ ID NO: 16, wherein CDR1, CDR2, and CDR3 are according to the Kabat numbering scheme. In some embodiments, the CD73 antibody comprises: (1) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 10 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 10; (2) LCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 11 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 11; and (3) LCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 12 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 12.

[0062] In some embodiments, the PD-1 antibody comprises a light chain variable region (VL) comprising an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:16.

[0063] In some embodiments, the PD-1 antibody comprises: (1) HCDR1, HCDR2 and HCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the heavy chain variable region (VH) as shown in SEQ ID NO: 17, and (2) LCDR1, LCR2 and LCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the light chain variable region (VL) as shown in SEQ ID NO: 16, wherein CDR1, CDR2 and CDR3 are according to the Kabat numbering scheme.

[0064] In some embodiments, the PD-1 antibody comprises: (1) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 13 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 13; (2) HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 14 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 14; (3) HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 15 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 15; (4) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 10 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 10; (5) LCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 11 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 11; and (6) LCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 12 or an amino acid sequence having one or more substitutions compared to SEQ ID NO: 12.

[0065] In some embodiments, the PD-1 antibody comprises: (1) a heavy chain variable region (VH) comprising an amino acid sequence as shown in SEQ ID NO: 17, or an amino acid sequence that is at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 17, and (2) a light chain variable region (VL) comprising an amino acid sequence as shown in SEQ ID NO: 16, or an amino acid sequence that is at least 80%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 16.

[0066] Exemplary anti-PD-1 antibodies that can be used in the methods of the present application include, but are not limited to, polyclonal antibodies, monoclonal antibodies, Fab, scFv, diantibodies, triantibodies, miniantibodies, VHHs, and sdAbs. Exemplary anti-PD-1 antibodies include, but are not limited to, pidilizumab, cemiplimab, sintilimab, cetrelimab, spartalizumab, camrelizumab, tislelizumab, balstilimab, toripalimab, dostarlimab, ABBV-181, penpulimab, Pembrolizumab, genolimzumab, retifanlimab. ), sasanlimab, AMP-224, AB122, F-520, MEDI-3387, MEDI-5771, MEDI-0680, SG-001, nivolumab, BCD-100, BAT-1306, BI-754091, CBT-501, GLS-010, LZM-009, Sym-021, CS-1003, HLX-10, AK-103, AM-0001, ENUM-244C8, ENUM-388D4, JTX-4014, RXI-762, STI-A1110, HLX-20, SSI-361, APL-501, TJ0141H, and SNA-01. In some embodiments, the anti-PD-1 antibody is toripalimab. method

[0067] In one aspect, a method for treating a solid tumor is provided, comprising administering a CD73 antibody or an antigen-binding fragment thereof to a subject in need thereof at a dose of about 2 to about 50 mg / kg body weight. In some embodiments, the method comprises administering a CD73 antibody to a subject in need thereof at a dose of about 10 to about 40 mg / kg body weight. In some embodiments, the method comprises administering a CD73 antibody to a subject in need thereof at a dose of about 20 to about 40 mg / kg body weight. In some embodiments, the method comprises administering a CD73 antibody to a subject in need thereof at a dose of about 25 to about 35 mg / kg body weight. In some embodiments, the method comprises administering a CD73 antibody as described herein to a subject in need thereof at a dose of about 30 mg / kg body weight.

[0068] In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 3 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 4 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 5 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 6 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 7 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 8 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 9 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 10 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 11 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 12 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 13 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 14 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 15 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 16 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 17 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 18 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered at a dose of about 19 to about 20 mg / kg body weight.

[0069] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 13 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 12 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 11 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 10 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 9 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 8 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 7 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 6 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 2 to about 5 mg / kg body weight.

[0070] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 13 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 12 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 11 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 10 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 9 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 8 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 7 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 6 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 3 to about 5 mg / kg body weight.

[0071] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 13 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 12 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 11 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 10 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 9 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 8 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 7 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 6 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 4 to about 5 mg / kg body weight.

[0072] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 13 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 12 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 11 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 10 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 9 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 8 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 7 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 5 to about 6 mg / kg body weight.

[0073] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 13 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 12 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 11 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 10 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 9 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 8 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 7 mg / kg body weight.

[0074] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 7 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 7 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 6 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 7 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 7 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 7 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 7 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 7 to about 13 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 7 to about 12 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 7 to about 11 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 7 to about 10 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 7 to about 9 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 7 to about 8 mg / kg body weight.

[0075] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 13 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 12 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 11 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 10 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 8 to about 9 mg / kg body weight.

[0076] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 13 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 12 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 11 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 9 to about 10 mg / kg body weight.

[0077] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 10 to about 13 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 10 to about 12 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 10 to about 11 mg / kg body weight.

[0078] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 11 to about 13 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 11 to about 12 mg / kg body weight.

[0079] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 14 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 12 to about 13 mg / kg body weight.

[0080] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 13 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 13 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 13 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 13 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 13 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 13 to about 15 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 13 to about 14 mg / kg body weight.

[0081] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 14 to about 20 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 14 to about 19 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 14 to about 18 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 14 to about 17 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 14 to about 16 mg / kg body weight. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered to a subject at a dose of about 14 to about 15 mg / kg body weight.

[0082] In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 15 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 15 to about 19 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 15 to about 18 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 15 to about 17 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 15 to about 16 mg / kg body weight.

[0083] In some embodiments, the CD73 antibody or antigen binding fragment thereof is administered to the subject at a dose of about 16 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or antigen binding fragment thereof is administered to the subject at a dose of about 16 to about 19 mg / kg body weight. In some embodiments, the CD73 antibody or antigen binding fragment thereof is administered to the subject at a dose of about 16 to about 18 mg / kg body weight. In some embodiments, the CD73 antibody or antigen binding fragment thereof is administered to the subject at a dose of about 16 to about 17 mg / kg body weight.

[0084] In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 17 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 17 to about 19 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 17 to about 18 mg / kg body weight.

[0085] In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 18 to about 20 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 18 to about 19 mg / kg body weight. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 19 to about 20 mg / kg body weight.

[0086] In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 10 to about 30 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 15 to about 30 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 20 to about 30 mg / kg body weight. In some embodiments, the CD73 antibody or its antigen-binding fragment is administered to the subject at a dose of about 25 to about 30 mg / kg body weight.

[0087] In some embodiments, the CD73 antibody or antigen binding fragment thereof is about 2, about 2.5, about 3, about 3.5, about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 8, about 8.5, about 9, about 9.5, about 10, about 10.5, about 11, about 11.5, about 12, about 12.5, about 13, about 13.5, about 14, about 14.5, about 15, about 15.5, about 16 , about 16.5, about 17, about 17.5, about 18, about 18.5, about 19, about 19.5, about 20 mg / kg, about 20.5, about 21, about 21.5, about 22, about 22.5, about 23, about 23.5, about 24, about 24.5, about 25, about 25.5, about 26, about 26.5, about 27, about 28.5, about 29, about 29.5, and about 30 mg / kg body weight are administered to a subject. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to a subject at a dose of about 20 mg / kg body weight.

[0088] In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to the subject once a week (QW), once every two weeks (Q2W), once every three weeks (Q3W), once every four weeks (Q4W), or once a month. In some embodiments, the CD73 antibody or antigen-binding fragment thereof is administered to the subject once every three weeks (Q3W).

[0089] In some embodiments, a CD73 antibody or antigen-binding fragment thereof is administered to a subject once every three weeks (Q3W) at a dose of 25-35, or 20 or 30 mg / kg.

[0090] In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered intravenously to a subject. In some embodiments, a CD73 antibody or an antigen-binding fragment thereof is administered intravenously to a subject once every three weeks (Q3W) at a dose of 20 or 30 mg / kg. In some embodiments, the CD73 antibody is uliledlimab, and uliledlimab is administered intravenously to a subject once every three weeks (Q3W) at a dose of 20 or 30 mg / kg. In some embodiments, the CD73 antibody is oleclumab, and oleclumab is administered intravenously to a subject once every three weeks (Q3W) at a dose of 20 or 30 mg / kg.

[0091] In some embodiments, the solid tumor is selected from the group consisting of acute lymphoblastic leukemia, multiple myeloma, glioma, mesothelioma, ovarian cancer, sarcoma, cervical cancer, bladder cancer, breast cancer, colorectal cancer, endometrial cancer, esophageal cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, lymphoma, melanoma, skin cancer (basal cell carcinoma), pancreatic cancer, prostate cancer, uterine cancer and thyroid cancer. In some embodiments, lung cancer includes non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC).

[0092] In some embodiments, the solid tumor is an advanced solid tumor. In some embodiments, the solid tumor is a metastatic solid tumor. In some embodiments, the solid tumor is a refractory solid tumor. In some embodiments, the solid tumor is in stage III or stage IV.

[0093] In some embodiments, the method further comprises administering a PD-1 antagonist to the subject.Any suitable PD-1 antagonist known in the art can be used in the present application.

[0094] In some embodiments, the PD-1 antagonist is a PD-1 antibody. In some embodiments, the PD-1 antagonist is a PD-1 antibody described herein. In some embodiments, the PD-1 antibody is selected from the group consisting of: Pembrolizumab, nivolumab, toripalimab, pidilizumab, cemiplimab, sintilimab, cetrelimab, spartalizumab, camrelizumab, tislelizumab, balstilimab, dostarlimab, ABBV-181, penpulimab, genolimzumab ), retifanlimab, sasanlimab, AMP-224, AB122, F-520, MEDI-3387, MEDI-5771, MEDI-0680, SG-001, BCD-100, BAT-1306, BI-754091, CBT-501, GLS-010, LZM-009, Sym-021, CS-1003, HLX-10, AK-103, AM-0001, ENUM-244C8, ENUM-388D4, JTX-4014, RXI-762, STI-A1110, HLX-20, SSI-361, APL-501, TJ0141H, and SNA-01. In some embodiments, the PD-1 antibody is toripalimab.

[0095] In some embodiments, the PD-1 antibody is toripalimab, and is administered to the subject at a fixed dose of 240 mg once every three weeks (Q3W).

[0096] In some embodiments, the method comprises administering to the subject intravenously a dose of 20 or 30 mg / kg of uliledlimab once every three weeks (Q3W) and a fixed dose of 240 mg of toripalimab once every three weeks (Q3W). In some embodiments, the method comprises administering to the subject intravenously a dose of 20 or 30 mg / kg of oleclumab once every three weeks (Q3W) and a fixed dose of 240 mg of toripalimab once every three weeks (Q3W).

[0097] In some embodiments, the antibody or antigen-binding fragment thereof and the PD-1 antagonist are administered simultaneously. In some embodiments, the antibody or antigen-binding fragment thereof and the PD-1 antagonist are administered separately.

[0098] In some embodiments, the method comprises administering to the subject intravenously at a dose of 20 or 30 mg / kg of uliledlimab once every three weeks (Q3W) and a fixed dose of 240 mg of toripalimab once every three weeks (Q3W). In some embodiments, the method comprises administering to the subject intravenously at a dose of 20 or 30 mg / kg of oleclumab once every three weeks (Q3W) and a fixed dose of 240 mg of toripalimab once every three weeks (Q3W).

[0099] In another aspect, the present application provides a use of a CD73 antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating a solid tumor in a subject in need thereof, wherein the medicament is administered to the subject in combination with a PD-1 antagonist at a dose of about 2 to about 30 mg / kg body weight.

[0100] In some embodiments, the subject has tumor tissue in which at least 40% of the tumor cells are CD73 positive. In some embodiments, the subject has tumor tissue in which at least 30%, 35%, 40%, 45%, or 50% of the tumor cells are CD73 positive. In some embodiments, at least 1% of the tumor cells are PD-L1 positive.

[0101] In some embodiments, the treatment methods disclosed herein require measuring CD73 expression in a sample from a patient, and administering a CD73 antibody or antigen-binding fragment thereof if at least 40% of the tumor cells in the sample are positive for CD73 expression. In some embodiments, the method also requires measuring PD-L1 expression in the sample.

[0102] In some embodiments, if at least 40% of the tumor cells are CD73 positive, a CD73 antibody or an antigen binding fragment thereof is administered to the subject. In some embodiments, if at least 30%, 35%, 40%, 45%, or 50% of the tumor cells are CD73 positive, a CD73 antibody or an antigen binding fragment thereof is administered to the subject. In some embodiments, if at least 1% of the tumor cells are PD-L1 positive, a CD73 antibody or an antigen binding fragment thereof is administered to the subject. In some embodiments, a PD-1 antagonist is further administered to the subject.

[0103] In some embodiments, CD73 expression is protein expression of CD73. In some embodiments, PD-L1 expression is protein expression of PD-L1. Protein levels can be determined or measured using any suitable method known in the art. In some embodiments, the method determines protein expressed on the cell membrane. In some embodiments, the method determines the level of expression in the cytosol. In some embodiments, the method determines the level of CD73 protein expressed on the cell membrane and in the cytosol.

[0104] In some embodiments, the protein expression level is determined by immunohistochemistry (IHC), which measures the amount of CD73 or PD-L1 protein in a cancer tissue sample (e.g., a NSCLC tissue sample). In some embodiments, the sample is a formalin-fixed and / or paraffin-embedded (FFPE) sample. In some embodiments, the sample is an archived sample. In some embodiments, the sample is a fresh sample. In some embodiments, the sample is a frozen sample. In some embodiments, the sample is obtained prior to the combined treatment described in the present application.

[0105] In some embodiments of the method of the present application, in IHC detection, according to the staining intensity, the level of CD73 protein in each cell (including cell membrane and cytosol) can be set to an intensity of 0, 1, 2 or 3 based on the standards shown in Table 1. Table 1. CD73 staining criteria Staining intensity standard 0 No staining 1 Faint or weak staining 2 Medium or moderate staining 3 Intense or strong staining

[0106] In some embodiments, the IHC assay determines the percentage of cells relative to the total cells with staining intensities 1, 2, and 3. In some embodiments, the cells include, but are not limited to, tumor cells, immune cells (e.g., tumor infiltrating immune cells (TILs)), stromal cells, vascular cells, and any combination thereof.

[0107] In some embodiments, the IHC assay determines the percentage of tumor cells with staining relative to total tumor cells. In some embodiments, the IHC assay determines the percentage of tumor cells with staining intensities 1, 2, and 3 relative to total tumor cells (TC). Products

[0108] In another aspect, the present application provides an article of manufacture comprising: (1) a CD73 antibody or an antigen-binding fragment thereof, and (2) a package insert suggesting that the CD73 antibody or antigen-binding fragment thereof be administered in combination with a PD-1 antagonist to a subject in need thereof at a dose of about 2 to about 20 mg / kg body weight. In some embodiments, the article of manufacture further comprises a PD-1 antagonist.

[0109] All publications and patent applications cited in this specification are incorporated herein by reference as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. Example

[0110] The following examples are presented to provide a person of ordinary skill in the art with a complete disclosure and description of how to make and use the invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are the inventors intended to represent that the following experiments are all or the only experiments performed. Efforts have been made to ensure the accuracy of the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should also be taken into account. Unless otherwise indicated, parts are by part weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure. Example 1. Phase I / II study of Uliledlimab for advanced solid tumors

[0111] A dose-escalation and dose-expansion Phase I / II study was designed to evaluate the safety, pharmacokinetics (PK), pharmacodynamics (PD), and preliminary efficacy of uliledlimab (ULI) alone or in combination with toripalimab (TOR) in patients with advanced solid tumors. ULI was administered intravenously at 2, 5, or 10 mg / kg weekly (QW), or at 15, 20, or 30 mg / kg every 3 weeks (Q3W), alone, or in combination with TOR (240 mg, intravenous injection, Q3W), with increasing doses. In the dose-expansion cohort, the preliminary efficacy of ULI (20 mg / kg, Q3W) + TOR (240 mg, Q3W) in non-small cell lung cancer (NSCLC) was studied. Serum concentrations of ULI and soluble CD73, and CD73 receptor occupancy (RO) in circulating CD19+ B cells were measured. Tumor response was assessed using RECIST1.1 / iRECIST.

[0112] A total of 92 patients were enrolled: 12 in the dose-escalation cohort of ULI alone, 9 in the ULI+TOR dose-escalation cohort, and 71 in the ULI+TOR dose-expansion cohort. ULI was well tolerated, and no DLTs were observed during dose escalation. Most treatment-related adverse events (TRAEs) were grade 1-2, while grade 3-4 TRAEs occurred in 25% of the ULI-alone group (2 patients had decreased lymphocyte counts and 1 patient had transient QT prolongation) and in 13.8% of the ULI+TOR group (the most commonly reported events were hypotension and diarrhea in 2 patients each). The most common TRAEs were grade 1-2 chills (47.8%), vomiting (46.7%), pyrexia (40.2%), diarrhea (32.6%), nausea (19.6%), pruritus (14.1%), and rash (13%), which occurred after the first dose of ULI and resolved with subsequent infusions and could be significantly mitigated by steroid-based premedication. At doses ≥5 mg / kg, the PK of ULI was linear, and the model showed a mean derived effective t1 / 2 of 13.2 days. After the first dose ≥15 mg / kg Q3W, the C 波谷 , soluble CD73 was undetectable and complete RO was maintained. Among the 48 NSCLC patients evaluable for efficacy in the dose expansion cohort, 6 partial responses (PR) (ORR 12.5%, 95% CI: 4.7%-25.2%) and 21 stable diseases (DCR 56.4%, 95% CI, 41.2%-70.5%) were observed. PR occurred in 3 of 8 patients who had not received systemic treatment, and PR occurred in 3 of 39 patients with relapsed / refractory disease to previous systemic treatment (including 2 patients who had not received previous PD-(L)1 inhibitor treatment and 1 patient who failed).

[0113] ULI was safe and tolerated up to 30 mg / kg, Q3W. Linear PK was observed at doses ≥5 mg / kg. The t1 / 2 was 13.2 days. At doses ≥15 mg / kg, Q3W, circulating and cell-bound CD73 were fully saturated. ULI combined with TOR showed evidence of antitumor activity in NSCLC patients. Example 2. Pharmacokinetic profile of Uliledlimab

[0114] This example performed a pharmacokinetic (PK) analysis in a trial that included NSCLC patients. ULI doses included 2 mg / kg (QW), 5 mg / kg (QW), 10 mg / kg (QW), 15 mg / kg (Q3W), 20 mg / kg (Q3W) and 30 mg / kg (Q3W).

[0115] At doses of 5 mg / kg weekly or higher, ULI exhibited a linear PK profile, indicating target saturation. The maximal inhibition of CD73 by ULI at C was estimated using relevant receptor occupancy (RO) data and ≥80% maximal inhibition of CD73 in tumor cell lines (with a reasonable assumption of 10% tumor penetration). 波谷 The target concentration at the time of the assay was 80 μg / mL in the hope of obtaining sufficient target inhibition.

[0116] ULI simulation in humans based on a population PK model established on clinical PK data ( Figure 1 ) shows that when C 波谷 Adequate tumor target binding can be expected at a concentration of 80 ug / mL. Thus, 30 mg / kg Q3W (with an additional dose on Cycle 1 Day 8 (C1D8)) can be considered an appropriate ULI dosing regimen.

[0117] To confirm the occupancy of CD73 receptor in tumors and the inhibitory effect of ULI on CD73 enzymatic activity at the selected dose of 30 mg / kg Q3W, 波谷 Tumor biopsies from two NSCLC patients treated with 30 mg / kg ULI were collected at time points C3D1 or C4D1. Total CD73 expression in tumors, including those bound to ULI, was determined by immunohistochemistry (IHC) staining using an anti-CD-73 antibody that does not compete with ULI ( Figure 2A and 2D ), and the expression of free CD73 not bound to ULI was determined by IHC staining using an anti-CD73 antibody that competes with ULI ( Figure 2B and 2E In addition, residual CD73 enzymatic activity in tumors was measured by enzyme histochemistry (EHC) staining ( Figure 2C and 2F At a dose of 30 mg / kg Q3W, ULI saturated CD73 occupancy, leading to complete inhibition of CD73 enzymatic activity in tumors, even in C 波谷 hour. Example 3. CD73 expression level is associated with response

[0118] This example evaluates CD73 expression levels in tumors and its correlation with patient response to treatment with ULI combined with toripalimab (TOR).

[0119] Based on IHC staining of tumor samples at baseline, patients were generally divided into high CD73 group and low CD73 group.Clinical response was measured by objective response rate (ORR) and disease control rate (DCR) according to iRECIST criteria.

[0120] To determine the appropriate CD73 cut-off level in predicting this clinical response, a receiver operating characteristic (ROC) curve analysis was performed on all treated patients (n=45). The ROC curve is presented in Figure 3 Based on this, a cutoff value of 40% was selected. Here, a value of 40% means that 40% of tumor or immune cells were stained with CD73 antibody (the staining intensity can be any of 1 (weak), 2 (moderate) or 3 (strong)).

[0121] As shown in Table 1, high CD73 patients had a significantly better response to treatment compared to low CD73 patients. Table 1. CD73 expression and response All patients (n=45#) High CD73 (n=18) Low CD73 (n=27) ORR 29％(13 / 45) 50％(9 / 18) 15％(4 / 27) DCR 73％(33 / 45) 89％(16 / 18) 63％(17 / 27)

[0122] When CD73 levels were combined with PD-L1 expression levels, their ability to stratify patients into high and low response groups was further improved. Figure 4 As shown in the results, in the PD-L1 1-49% group and the PD-L1 ≥ 50% group, the clinical response improvement of patients with high CD73 was more prominent than that of patients with low CD73 compared with that of the PD-L1 < 1% group.

[0123] The present invention has been described to include the preferred mode of the present invention's practice based on the specific embodiments found or proposed by the inventors. It will be appreciated by those skilled in the art that, according to the present application disclosure, a large number of modifications and changes can be made to the specific embodiments of the examples without departing from the intended scope of the present invention. For example, due to codon redundancy, potential DNA sequences can be changed without affecting the protein sequence. In addition, for the consideration of biological function equivalence, protein structure can be changed without affecting its biological effect in kind or quantity. All these modifications are expected to be included in the scope of the appended claims. Sequence Listing

Claims

1. A method for treating solid tumors, comprising: The CD73 antibody or antigen-binding fragment thereof and the PD-1 antagonist are administered to a subject in need thereof at a dose of about 2 to about 60 mg / kg body weight.

2. The method according to claim 1, wherein: The CD73 antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 10 to about 40 mg / kg body weight.

3. The method according to claim 1, wherein: The CD73 antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 25 to about 35 mg / kg body weight.

4. The method according to claim 1, wherein: The CD73 antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 30 mg / kg body weight.

5. The method according to any one of claims 1 to 4, wherein: The CD73 antibody or antigen-binding fragment thereof is administered to the subject once a week (QW), once every two weeks (Q2W), once every three weeks (Q3W), once every four weeks (Q4W), or once a month.

6. The method according to any one of claims 1 to 5, wherein: The CD73 antibody or antigen-binding fragment thereof is administered to the subject once every three weeks (Q3W).

7. The method according to any one of claims 1 to 6, wherein: The CD73 antibody or antigen-binding fragment thereof is administered to the subject at a dose of 25 or 35 mg / kg once every three weeks (Q3W).

8. The method according to any one of claims 1 to 7, wherein: The CD73 antibody or antigen-binding fragment thereof is administered intravenously to the subject.

9. The method according to any one of claims 1 to 8, wherein: The CD73 antibody or antigen-binding fragment thereof is selected from the group consisting of full-length antibody, Fab, Fab', F(ab')2, scFv and sdAb.

10. The method according to claim 9, wherein: The CD73 antibody or antigen-binding fragment thereof is a full-length antibody.

11. The method according to any one of claims 1 to 10, wherein: The CD73 antibody or antigen-binding fragment thereof comprises HCDR1, HCDR2 and HCDR3, which respectively comprise the amino acid sequences of CDR1, CDR2 and CDR3 in the heavy chain variable region (VH) as shown in SEQ ID NO: 1, wherein CDR1, CDR2 and CDR3 are according to the Kabat numbering scheme.

12. The method according to any one of claims 1 to 11, wherein: The CD73 antibody or antigen-binding fragment thereof comprises: (1) HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 3, (2) HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 4, (3) HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 5, (4) LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 6, (5) LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 7, and (6) LCDR3 comprising the amino acid sequence shown in SEQ ID NO:

8.

13. The method according to any one of claims 1 to 12, wherein: The CD73 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region (VH) comprises the amino acid sequence as shown in SEQ ID NO: 1, and the light chain variable region (VL) comprises the amino acid sequence as shown in SEQ ID NO:

2.

14. The method of any one of claims 1-13, wherein the CD73 antibody or antigen-binding fragment thereof is selected from the group consisting of uliledlimab (I-Mab Biopharma), oleclumab (AstraZeneca), CPI-006 (Corvus Pharma), BMS-986179 (Bristol-Myers Squibb), AB-680 (Arcus Biosciences), NZV-930 (SRF373, Surface Oncolohgy / Novartis), JAB-BX102 (Jacobio), AK119 (Akesobio), Sym024 (Symphogen), IBI 325 (Innovent), BR 101 (HisunBioRay) and LY3475070 (Eli Lilly).

15. The method according to any one of claims 1 to 14, wherein: The solid tumor is selected from the group consisting of ovarian cancer, sarcoma, cervical cancer, bladder cancer, breast cancer, colorectal cancer, endometrial cancer, esophageal cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, lymphoma, acute lymphoblastic leukemia, multiple myeloma, glioma, mesothelioma, melanoma, skin cancer (basal cell carcinoma), pancreatic cancer, prostate cancer, uterine cancer and thyroid cancer.

16. The method according to any one of claims 1 to 15, wherein: The lung cancer includes non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC).

17. The method according to any one of claims 1 to 16, wherein: The solid tumor is an advanced solid tumor.

18. The method according to any one of claims 1 to 17, wherein: The solid tumor is a metastatic solid tumor.

19. The method according to any one of claims 1 to 18, wherein: The solid tumor is a recurrent solid tumor.

20. The method according to any one of claims 1 to 19, wherein: The solid tumor is a refractory solid tumor.

21. The method according to any one of claims 1 to 20, wherein: The solid tumor is in stage III or stage IV.

22. The method according to any one of claims 1 to 21, wherein: The CD73 antibody or antigen-binding fragment thereof and the PD-1 antagonist are administered simultaneously.

23. The method according to any one of claims 1 to 21, wherein: The CD73 antibody or antigen-binding fragment thereof and the PD-1 antagonist are administered separately.

24. The method according to any one of claims 1 to 23, wherein: The PD-1 antagonist is a PD-1 antibody.

25. The method according to claim 24, wherein: The PD-1 antibody comprises: (1) LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 10, (2) LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 11, (3) LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 12, (4) HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 13, (5) HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 14, and (6) HCDR3 comprising the amino acid sequence shown in SEQ ID NO:

15.

26. The method according to claim 24 or 25, wherein: The PD-1 antibody comprises: (1) a heavy chain variable region (VH) comprising an amino acid sequence as shown in SEQ ID NO: 16 or an amino acid sequence having at least 90% identity with SEQ ID NO: 16, and (2) a light chain variable region (VL) comprising an amino acid sequence as shown in SEQ ID NO: 17 or an amino acid sequence having at least 90% identity with SEQ ID NO:

17.

27. The method according to claim 24, wherein: The PD-1 antibody is selected from the group consisting of: pembrolizumab, nivolumab, toripalimab, pidilizumab, cemiplimab, sintilimab, cetrelimab, spartalizumab, camrelizumab, tislelizumab, balstilimab, dostarlimab, ABBV-181, penpulimab, genolimzumab, retifanlimab, sasanlimab, AMP-224, AB122, F-520, MEDI-3387, MEDI-5771, MEDI-0680, SG-001, BCD-100, BAT-1306, BI-754091, CBT-501, GLS-010, LZM-009, Sym-021, CS-1003, HLX-10, AK-103, AM-0001, ENUM-244C8, ENUM-388D4, JTX-4014, RXI-762, STI-A1110, HLX-20, SSI-361, APL-501, TJ0141H, and SNA-01.

28. The method according to claim 27, wherein: The PD-1 antibody is Teplizumab.

29. The method according to claim 28, wherein: The PD-1 antibody was administered to the subject at a fixed dose of 240 mg once every three weeks (Q3W).

30. The method according to any one of claims 1 to 29, wherein: The subject has tumor tissue in which at least 40% of the tumor cells are CD73 positive.

31. The method according to any one of claims 1 to 29, wherein: The subject has tumor tissue in which at least 30%, 35%, 40%, 45% or 50% of the tumor cells are CD73 positive.

32. The method according to claim 30 or 31, wherein: At least 1% of tumor cells are PD-L1 positive.

33. Use of a CD73 antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating a solid tumor in a subject in need thereof, wherein the medicament is administered to the subject in combination with a PD-1 antagonist at a dose of about 2 to about 60 mg / kg body weight.

34. An article comprising: (1) a CD73 antibody or an antigen-binding fragment thereof, and (2) a package insert that recommends administering the CD73 antibody or antigen-binding fragment thereof in combination with a PD-1 antagonist to a subject in need thereof at a dose of about 2 to about 60 mg / kg body weight.

35. The article of manufacture according to claim 34, further comprising a PD-1 antagonist.

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