Combination Therapies Containing KRAS Inhibitors for the Treatment of Cancer - Patent application

Abstract

Provided herein is a method for treating KRAS mutant cancer (e.g., KRAS G12C mutant cancer or KRAS G12D mutant cancer) in a subject, comprising administering to the subject an effective amount of an agent that blocks the interaction between CD47 and SIRPα and an effective amount of a KRAS inhibitor (e.g., when the cancer is a KRAS G12C mutant cancer, a KRAS G12C inhibitor, or when the cancer is a KRAS G12D mutant cancer, a KRAS G12D inhibitor).Also provided is a method for stimulating the phagocytosis of a population of cancer cells that express KRAS mutant protein (e.g., KRAS G12C mutant protein or KRAS G12D mutant protein) by macrophages.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to International Application No. PCT / CN2022 / 079307, filed March 4, 2022, the contents of which are incorporated by reference in their entirety herein.

[0002] Reference to Electronic Sequence Listing The contents of the electronic format of the sequence listing (233002000741SEQLIST.xml, size: 45,965 bytes, and creation date: February 15, 2023) are incorporated by reference in their entirety into this specification.

[0003] The present application relates to combination therapies for the treatment of KRAS mutant cancers, such as non-small cell lung cancer (NSCLC), colon cancer, colorectal cancer, and the like. [Background technology]

[0004] KRAS mutations play a role in some of the most common and deadly cancers, including lung, colon, colorectal, and rectal cancers. G12C is a single point mutation with a glycine to cysteine ​​substitution at codon 12 of the KRAS protein. This substitution favors an active, GTP-bound conformation of KRAS, which amplifies the signaling pathway that leads to carcinogenesis. KRAS G12C is particularly prevalent in non-small cell lung cancer (NSCLC), which comprises approximately 85% of all lung cancer cases in the United States. Approximately 13% of Americans with NSCLC have the KRAS G12C mutation, and approximately 23,000 new cases of KRAS G12C NSCLC are diagnosed each year in the United States alone. Clinical trials of the KRAS G12C allele-specific inhibitors adagrasib and sotorasib have shown promising activity in cancers expressing the KRAS G12C mutant protein. However, clinical trial data also show that there is a large variability in response among patients treated with KRAS G12C inhibitors, and that monotherapy with KRAS G12C inhibitors is unlikely to be sufficient to elicit sustained therapeutic response.G12D is another single point mutation with a glycine to aspartic acid substitution at codon 12 of KRAS protein.G12D mutation accounts for about 41% of all G12 mutations, and is the most common G12 mutation in colorectal cancer and pancreatic ductal adenocarcinoma (PDAC).Therefore, there is a continuing need in the art for a method of treating cancers expressing KRAS G12C mutant protein or KRAS G12D mutant protein.

[0005] All references cited herein, including patent applications, patent publications, and UniProtKB / Swiss-Prot accession numbers, are hereby incorporated by reference in their entirety, as if each individual reference was specifically and individually indicated to be incorporated by reference. Summary of the Invention [Means for solving the problem]

[0006] Provided herein is a method of treating cancer in a subject, comprising administering to the subject an effective amount of an agent that blocks the interaction between CD47 and SIRPα and an effective amount of a KRAS inhibitor, wherein the cancer comprises one or more cancer cells that express a KRAS mutant protein.Also provided herein is a method of stimulating phagocytosis of a population of cancer cells by macrophages, comprising contacting the population with an effective amount of an agent that blocks the interaction between CD47 and SIRPα and an effective amount of a KRAS inhibitor, wherein the population of cancer cells comprises one or more cancer cells that express a KRAS mutant protein.

[0007] In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is a polypeptide that binds to CD47.In some embodiments, the polypeptide that binds to CD47 is an anti-CD47 antibody or an immunologically active fragment thereof.In some embodiments, the anti-CD47 antibody or an immunologically active fragment thereof is CC-90002, 5F9, LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or any one of the immunologically active fragments of the above.

[0008] In some embodiments, the anti-CD47 antibody or immunologically active fragment thereof comprises a heavy chain variable domain (V H ) and the three complementarity determining regions (CDRs) of the light chain variable domain (V L In some embodiments, the anti-CD47 antibody or immunologically active fragment thereof comprises three CDRs: (a) V, comprising (1) a CDR-H1 comprising RAWMN (SEQ ID NO: 5), (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO: 6), and (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 7). Hand (b) (1) a CDR-L1 comprising KSSQSVLYAGNNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10), L and wherein the CDR sequences are defined according to the Kabat numbering system. In some embodiments, the V of an anti-CD47 antibody or immunologically active fragment thereof comprises H The domain comprises an amino acid sequence having at least 95% identity to SEQ ID NO:1 and is a V domain of an anti-CD47 antibody or an immunologically active fragment thereof. L comprises an amino acid sequence having at least 95% identity to SEQ ID NO:2. H The N-terminal amino acid of the domain is E, and V H The C-terminal amino acid of the domain is S. In some embodiments, (a) the C-terminal amino acid of the domain is V according to the Kabat numbering system. H The N-terminal amino acid of the domain corresponds to position H1 and V H The C-terminal amino acid of the domain corresponds to position H113. (b) According to the Chothia numbering system, V H The N-terminal amino acid of the domain corresponds to position H1 and V H The C-terminal amino acid of the domain corresponds to position H113, or (c) V according to the IMGT numbering system. H The N-terminal amino acid of the domain corresponds to position H1 and V H The C-terminal amino acid of the domain corresponds to position H128. H The N-terminal amino acid of the domain corresponds to amino acid 1 of SEQ ID NO:1, H The C-terminal amino acid of the domain corresponds to amino acid 118 of SEQ ID NO:1. In some embodiments, the anti-CD47 antibody is a full length antibody. In some embodiments, the full length anti-CD47 antibody comprises a human IgG4 Fc region comprising a S228P substitution, or a variant thereof, where the amino acid numbering is according to the EU index. In some embodiments, the full length anti-CD47 antibody comprises a heavy chain comprising SEQ ID NO:3 or SEQ ID NO:35, and a light chain comprising SEQ ID NO:4.

[0009] In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor, and the KRAS mutant protein is a KRAS G12C mutant protein. In some embodiments, the KRAS G12C inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of the KRAS G12C mutant protein. In some embodiments, the KRAS G12C inhibitor is a small molecule. In some embodiments, the small molecule is selected from the group consisting of AMG 510, MRTX849, JAB-21822, GDC-6036, JDQ443, D-1553, GH35, GFH925, BPI-421286, and LY3537982. In some embodiments, the small molecule is AMG 510 or MRTX849.

[0010] In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor, and the KRAS mutant protein is a KRAS G12D mutant protein.In some embodiments, the KRAS G12D inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of the KRAS G12D mutant protein.In some embodiments, the KRAS G12D inhibitor is a small molecule.In some embodiments, the small molecule is MTRX1122.

[0011] In some embodiments of any of the methods herein, the cancer is lung cancer, colon cancer, colorectal cancer, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric-esophageal cancer, or rectal cancer. In some embodiments, the cancer is KRAS G12C cancer. In some embodiments, the KRAS G12C cancer is lung cancer. In some embodiments, the lung cancer is lung adenocarcinoma or non-small cell lung cancer (NSCLC). In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC). In some embodiments, the cancer is KRAS G12D cancer. In some embodiments, the KRAS G12D cancer is colorectal cancer or colon cancer.

[0012] In some embodiments of any of the methods herein, the anti-CD47 antibody or immunologically active fragment thereof and the KRAS inhibitor are administered simultaneously. In some embodiments, the anti-CD47 antibody or immunologically active fragment thereof and the KRAS inhibitor are administered sequentially. In some embodiments, the anti-CD47 antibody or immunologically active fragment thereof is administered before the KRAS inhibitor. In some embodiments, the KRAS inhibitor is administered before the anti-CD47 antibody or immunologically active fragment thereof.

[0013] In some embodiments, the subject is a human.

[0014] In some embodiments of the methods herein, the one or more cancer cells expressing the KRAS mutant protein are lung cancer cells, colon cancer cells, colorectal cancer cells, pancreatic cancer cells, cholangiocarcinoma cells, endometrial cancer cells, ovarian cancer cells, peritoneal cancer cells, bladder cancer cells, gastric cancer cells, thyroid cancer cells, melanoma cells, breast cancer cells, head and neck cancer cells, multiple myeloma cells, acute myeloid leukemia (AML) cells, uterine cancer cells, gastric and esophageal cancer cells, or rectal cancer cells. In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor, the one or more cancer cells expressing the KRAS mutant protein express a KRAS G12C mutant protein, the one or more cancer cells expressing the KRAS G12C mutant protein are lung cancer cells, and the lung cancer cells are lung adenocarcinoma cells or non-small cell lung cancer (NSCLC) cells. In some embodiments, the lung cancer cells are NSCLC cells. In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor, the one or more cancer cells expressing a KRAS mutant protein express a KRAS G12D mutant protein, and the one or more cancer cells expressing a KRAS G12D mutant protein are colon cancer cells or colorectal cancer cells.

[0015] In some embodiments, a kit for treating cancer in a human subject is provided, comprising: (a) an agent that blocks the interaction between CD47 and SIRPα; and (b) instructions for administering an effective amount of the agent and an effective amount of a KRAS inhibitor to a subject having a cancer comprising one or more cancer cells expressing a KRAS mutant protein. In some embodiments, the polypeptide is a polypeptide that binds to CD47. In some embodiments, the polypeptide that binds to CD47 is an anti-CD47 antibody or an immunologically active fragment thereof. In some embodiments, the anti-CD47 antibody or an immunologically active fragment thereof is an anti-CD47 antibody or an immunologically active fragment thereof comprising: (a) a CDR-H1 comprising (1) RAWMN (SEQ ID NO: 5), (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO: 6), and (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 7). H and (b) (1) a CDR-L1 comprising KSSQSVLYAGNNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10), L and wherein the CDR sequences are defined according to the Kabat numbering system. In some embodiments, the V of an anti-CD47 antibody or immunologically active fragment thereof comprises H The domain comprises an amino acid sequence having at least 95% identity to SEQ ID NO:1 and is a V domain of an anti-CD47 antibody or an immunologically active fragment thereof. L comprises an amino acid sequence having at least 95% identity to SEQ ID NO:2. H The N-terminal amino acid of the domain is E, and V HThe C-terminal amino acid of the domain is S. In some embodiments, the anti-CD47 antibody is a full-length antibody. In some embodiments, the full-length anti-CD47 antibody comprises a heavy chain comprising SEQ ID NO:3 or SEQ ID NO:35 and a light chain comprising SEQ ID NO:4. In some embodiments, the kit further comprises a KRAS inhibitor. In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor and the cancer comprises one or more cancer cells expressing a KRASG12C mutant protein. In some embodiments, the KRAS G12C inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of the KRAS G12C mutant protein. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the small molecule is selected from the group consisting of AMG 510, MRTX849, JAB-21822, GDC-6036, JDQ443, D-1553, GH35, GFH925, BPI-421286, and LY3537982. In some embodiments, the small molecule is AMG 510 or MRTX849. In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor, and the cancer comprises one or more cancer cells expressing a KRASG12D mutant protein. In some embodiments, the KRAS G12D inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of a KRAS G12D mutant protein. In some embodiments, the KRAS G12D inhibitor is a small molecule inhibitor. In some embodiments, the small molecule is MRTX1133. In some embodiments, the cancer comprising one or more cancer cells expressing a KRAS mutant protein is lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal adenocarcinoma.

[0016] In some embodiments, a pharmaceutical composition is provided that includes an agent that blocks the interaction between CD47 and SIRPα and a KRAS inhibitor. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody or an immunologically active fragment thereof. In some embodiments, the anti-CD47 antibody or immunologically active fragment thereof is CC-90002, 5F9, LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment thereof. In some embodiments, the anti-CD47 antibody or immunologically active fragment thereof is selected from the group consisting of: (a) a CDR-H1 comprising (1) RAWMN (SEQ ID NO:5), (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO:6), and (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO:7). H and (b) (1) a CDR-L1 comprising KSSQSVLYAGNNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10), L and wherein the CDR sequences are defined according to the Kabat numbering system. In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor. In some embodiments, the KRAS G12C inhibitor is selected from the group consisting of AMG 510, MRTX849, JAB-21822, GDC-6036, JDQ443, D-1553, GH35, GFH925, BPI-421286, and LY3537982. In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor. In some embodiments, the KRAS G12D inhibitor is MTRX1133.

[0017] It should be understood that one, some, or all of the features of the various embodiments described herein may be combined to form other embodiments of the present invention. These and other aspects of the present invention will be apparent to those skilled in the art. These and other embodiments of the present invention are further described in the following detailed description. [Brief description of the drawings]

[0018] [Figure 1] 1 provides the results of an experiment conducted to compare the efficacy of (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, and (d) AMG-510 in combination with TJC4 in inhibiting tumor growth in mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts. The NCI-H358 cell line harbors a KRAS G12C mutation.

[0019] [Diagram 2] 1 provides an experiment performed to compare tumor weights from mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts following treatment with (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4.

[0020] [Figure 3A] 1 provides the results of flow cytometry experiments performed to determine levels of calreticulin ("CALR") in tumors from mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts following treatment with (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4.

[0021] [Figure 3B] 1 provides the results of flow cytometry experiments performed to determine the ratio of M1:M2 macrophages in tumors from mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts following treatment with (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4.

[0022] [Figure 3C]13 provides the results of flow cytometry experiments performed to determine the percentage of M1 macrophages in tumors from mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts following treatment with (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4.

[0023] [Figure 3D] 1 provides the results of multiplex immunohistochemistry experiments performed to determine the percentages of total, M1, and M2 macrophages in tumors from mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts following treatment with (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4.

[0024] [Figure 3E] 1 provides the results of multiplex immunohistochemistry experiments performed to determine the ratio of M1:M2 macrophages as determined via multiplex immunohistochemistry in tumors from mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts following treatment with (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4.

[0025] [Figure 3F] 13A-13D provide multiplex immunofluorescence staining images of tumors from mice bearing human NCI-H358 non-small cell lung cancer tumor xenografts following treatment with (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4.

[0026] [Figure 4A]1 provides the results of an in vitro phagocytosis assay performed using NCI-H358 cells incubated with M0 macrophages, 1 μg / ml TJC4, and 0.1 μm AMG 150.

[0027] [Figure 4B] 1 provides the results of an in vitro phagocytosis assay performed using NCI-H358 cells incubated with M0 macrophages, 1 μg / ml TJC4, and 1 μM AMG 150.

[0028] [Figure 4C] 1 provides the results of an in vitro phagocytosis assay performed using NCI-H358 cells incubated with M0 macrophages, 1 μg / ml TJC4, and 10 μm AMG 150.

[0029] [Figure 5A] 13 provides the results of flow cytometry experiments performed to evaluate the effects of DMSO, MRTX1133, TJC4, and MRTX1133+TJC4 on the phagocytosis of human LS1780 colon cancer cells by macrophages in vitro.

[0030] [Figure 5B] Quantitative results of the data shown in FIG. 5A are provided.

[0031] [Figure 6A] 1 provides the results of flow cytometry experiments performed to evaluate the effects of DMSO, MRTX1133, TJC4, and MRTX1133+TJC4 on apoptosis of human LS1780 colon cancer cells in vitro.

[0032] [Figure 6B] We provide quantitative results for the data shown in FIG. 6A.

[0033] [Figure 7A] 1 provides an overview of the experimental procedures described in Example 4.

[0034] [Figure 7B] 1 provides the results of experiments performed to evaluate the antitumor effects of PBS, MRTX1133, TJC4, and MRTX1133+TJC4 in mice bearing human LS180 colon cancer tumor xenografts.

[0035] [Figure 7C] 13 provides luciferase images of human LS180 colon cancer tumor xenografts in mice treated with PBS, MRTX1133, TJC4, or MRTX1133+TJC4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] definition Before describing the embodiments in detail, it is to be understood that this disclosure is not limited to particular compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0037] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a molecule" optionally includes a combination of two or more such molecules, and so forth.

[0038] The term "about" as used herein refers to a normal range of error for the respective value, which is readily apparent to one of ordinary skill in the art. Reference herein to "about" a value or parameter includes (and describes) embodiments that are directed to the value or parameter itself.

[0039] Aspects and embodiments of the present disclosure are understood to include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0040] The term "CD47", which is also known as integrin-associated protein (IAP), antigen surface determinant protein OA3, OA3, CD47 antigen, Rh-associated antigen, integrin-associated signaling factor, antigen identified by monoclonal antibody 1D8, CD47 glycoprotein, preferably refers to human CD47, and in particular to the amino acid sequence [ka] or a protein comprising said amino acid sequence variants. The term "CD47" also refers to any post-translationally modified and conformational variants.

[0041] As used herein, the term "antibody" is used in the broadest sense and specifically encompasses intact antibodies (e.g., full-length antibodies), antibody fragments (including but not limited to Fab, F(ab')2, scFv, scFv-Fc, single domain antibodies), monoclonal antibodies, and polyclonal antibodies, so long as they exhibit the desired biological activity (e.g., epitope binding).

[0042] 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 is one that has been identified and separated and / or recovered from components of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses of the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. An antibody may be rendered substantially free of naturally associated components (or components associated with the cellular expression system used to produce the antibody) by isolation using protein purification techniques well known in the art. In some embodiments, the antibody is purified to (1) greater than 75% by weight, most preferably greater than 80%, 90%, 95%, or 99% by weight of antibody as determined by the Lowry method, or (2) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue, preferably silver staining. Isolated antibody includes the antibody in situ within a recombinant cell, 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.

[0043] As used herein, the term "epitope" refers to any antigenic determinant on an antigen to which the paratope of an antibody binds. Epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics.

[0044] As used herein, the term "natural antibodies and immunoglobulins" refers to heterotetrameric glycoproteins of about 150,000 daltons, usually composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond (also referred to as a "VH / VL pair"), while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes. 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 several constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at the other end, with the constant domain of the light chain aligned with the first constant domain of the heavy chain, and the light chain variable domain aligned with the variable domain of the heavy chain. It is believed that certain amino acid residues form an interface between the light chain variable domain and the heavy chain variable domain. See, e.g., Chothia et al., J. Mol. Biol., 186:651 (1985); Novotny and Haber, Proc. Natl. Acad. Sci. USA, 82:4592 (1985).

[0045] As used herein, the term "variable" refers to the fact that the sequences of certain portions of the variable domains vary widely among antibodies and are used in the binding and specificity of each particular antibody to its particular antigen. However, the variability is not uniformly distributed throughout the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) or hypervariable regions in both the light and heavy chain variable domains. The more highly conserved portions of the variable domains are called frameworks (FRs). Natural heavy and light chain variable domains each contain four FR regions that are largely in a β-sheet configuration, and these FRs are connected by three CDRs that form loops that connect, and in some cases form part of, the β-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, together with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies. See, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991). The constant domains are not directly involved in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity. Variable region sequences of interest include the humanized variable region sequences of the CD47 antibody detailed elsewhere herein.

[0046] The term "hypervariable region (HVR)" or "complementarity determining region (CDR)" may refer to subregions of the VH and VL domains characterized by enhanced sequence variability and / or the formation of defined loops. These include three CDRs in the VH domain (H1, H2, and H3) and three CDRs in the VL domain (L1, L2, and L3). H3 is thought to be essential for conferring fine binding specificity, while L3 and H3 show the highest level of diversity. See Johnson and Wu, in Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, NJ, 2003).

[0047] Several CDR / HVR demarcations are known. The Kabat complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). Chothia instead refers to the location of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). The AbM HVRs represent a compromise between the Kabat HVRs and the Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. The "contact" HVRs are based on analysis of available complex crystal structures. Residues from each of these HVR / CDRs are listed below. "Framework" or "FR" residues are variable domain residues other than HVR / CDR residues. [Table 3]

[0048] "Extended" HVRs are also known: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL, and 26-35 (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in VH (Kabat numbering).

[0049] "Numbering according to Kabat" may refer to the numbering system used for the heavy or light chain variable domains of the collection of antibodies in Kabat et al. (supra). The actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion into, the FRs or HVRs of the variable domain. The Kabat numbering of residues may be determined for a given antibody by alignment of the antibody sequence at the regions of homology with a "standard" Kabat numbered sequence. Typically, Kabat numbering is used when referring to residues in the variable domain (approximately residues 1-107 for the light chain and residues 1-113 for the heavy chain), while the EU numbering system or index (e.g., EU index as in Kabat's numbering according to EU IgG1) is generally used when referring to residues in the heavy chain constant region.

[0050] As used herein, a "monoclonal" antibody refers to a substantially homogeneous antibody, e.g., an antibody obtained from a population of antibodies that are substantially identical but tolerate minor levels of background mutations and / or modifications. "Monoclonal" refers to the substantially homogeneous nature of the antibody and does not require the production of the antibody by any particular method. In some embodiments, a monoclonal antibody is selected for its HVR, VH, and / or VL sequence and / or binding characteristics, e.g., selected from a pool of clones (e.g., recombinant, hybridoma, or phage derived). A monoclonal antibody may be engineered to contain one or more mutations, e.g., to affect the binding affinity or other properties of the antibody, to create a humanized or chimeric antibody, to improve antibody production and / or homogeneity, to design a multispecific antibody, and the resulting antibody is still considered to be monoclonal in nature. A population of monoclonal antibodies may be distinguished from a polyclonal antibody because each monoclonal antibody in the population recognizes the same antigenic site. Various techniques for producing monoclonal antibodies are known, such as the hybridoma method (e.g., Kohler and Milstein, Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14(3):253-260 (1995); Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, NY, 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567), phage display technology (e.g., Clackson et al., Nature, 352:624-628 (1991); Marks et al., Nature, 352:624-628 (1991)), and the like. al., J.Mol.Biol.222:581-597(1992), Sidhu et al., J.Mol.Biol.338(2):299-310(2004), Lee et al., J. Mol. Biol. 340(5):1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34):12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2):119-132 (2004)), and techniques for producing human or human-like antibodies in animals having some or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (e.g., WO 1998 / 24893, WO 1996 / 34096, WO 1996 / 33735, WO 1991 / 10741, Jakobovits et al., Proc. Natl. Acad. Sci. USA 101(34):12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2):119-132 (2004)). 90:2551(1993), Jakobovits et al., Nature 362:255-258(1993), Bruggemann et al., Year in Immunol.7:33 (1993), U.S. Patent Nos. 5,545,807, 5,545,806, 5,569,825, 5,625,126, 5,633,425, and 5,661,016, Marks et al.,Bio / Technology 10:779-783(1992), Lonberg et al., Nature 368:856-859(1994), Morrison, Nature 368:812-813(1994), Fishwild et al., Nature Biotechnol. 14:845-851 (1996); Neuberger, Nature Biotechnol. 14:826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13:65-93 (1995).

[0051] A "chimeric" antibody may refer to an antibody having some portions of the heavy and / or light chains from a particular isotype, class, or organism, and other portions from another isotype, class, or organism, hi some embodiments, the variable regions are from one source or organism and the constant regions are from another source or organism.

[0052] A "humanized antibody" may refer to an antibody that has primarily human sequences and minimal amounts of non-human (e.g., mouse or chicken) sequences. In some embodiments, a humanized antibody has one or more HVR sequences (with the desired binding specificity) from an antibody derived from a non-human (e.g., mouse or chicken) organism grafted onto the framework (FR) of a human recipient antibody. In some embodiments, non-human residues are further grafted onto the human framework (not present in either the source or recipient antibody), e.g., to improve the properties of the antibody. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. A humanized antibody will also optionally comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. See Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).

[0053] A "human" antibody may refer to an antibody having an amino acid sequence that corresponds to that of an antibody produced by a human and / or an antibody made using any of the techniques for making human antibodies as disclosed herein. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991)), preparation of human monoclonal antibodies (such as those described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991)), and transgenic animals that have been engineered to produce such antibodies in response to antigenic challenge, but in which the endogenous locus has been disabled, e.g., immunized xenogeneic mice. xenomice (see, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584 for XENOMOUSE™ technology) or by administering antigen to chickens with human immunoglobulin sequence(s) (see, e.g., WO2012162422, WO2011019844, and WO2013059159).

[0054] There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, IgA2. The heavy chain constant domains that correspond to these different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known.

[0055] As used herein, the term "antibody fragment," and all grammatical variations thereof, is defined as a portion of an intact antibody that contains the antigen binding site or variable region of the intact antibody, which in certain cases does not contain the constant heavy chain domains of the Fc region of the intact antibody (i.e., CH2, CH3, and / or CH4, depending on the antibody isotype). Examples of antibody fragments include Fab, Fab', Fab'-SH, F(ab')2, and Fv fragments; diabodies; any antibody fragment which is a polypeptide having a primary structure consisting of one uninterrupted sequence of contiguous amino acid residues (referred to herein as a "single-chain antibody fragment" or "single-chain polypeptide"), including, but not limited to, (1) a single-chain Fv (scFv) molecule, (2) a single-chain polypeptide comprising only one light chain variable domain without associated heavy chain portions, or a fragment thereof comprising the three CDRs of the light chain variable domain, and (3) a single-chain polypeptide comprising only one heavy chain variable region without associated light chain portions, or a fragment thereof comprising the three CDRs of the heavy chain variable region; as well as multispecific or multivalent structures formed from antibody fragments. In antibody fragments comprising one or more heavy chains, the heavy chain(s) can comprise any of the constant domain sequences found in the non-Fc region of an intact antibody (e.g., CH1 in an IgG isotype) and / or can comprise any of the hinge region sequences found in an intact antibody and / or can comprise a leucine zipper sequence fused or positioned to the hinge region or constant domain sequence of the heavy chain(s).

[0056] Papain digestion of an antibody results in two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, a name reflecting the ability to crystallize readily. Pepsin treatment produces an F(ab')2 fragment that has two antigen-binding sites and is still capable of cross-linking antigen. "Fv" is the smallest antibody fragment that contains an intact antigen recognition and binding site. In two-chain Fv species, this region consists of a dimer of one heavy-chain variable domain and one light-chain variable domain in tight, non-covalent association. In single-chain Fv species (scFv), one heavy-chain variable domain and one light-chain variable domain can be covalently linked by a flexible peptide linker such that the light and heavy chains can associate in a "dimeric" structure similar to that in the two-chain Fv species. In this configuration, the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three antigen-specific CDRs) has the ability to recognize and bind antigen, although with lower affinity than the entire binding site. See, e.g., Pluckthun, in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0057] Fab fragments also contain the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CH1 domain including one or more cysteines from the antibody hinge region. Fab'-SH is the designation used herein to represent Fab' in which the cysteine ​​residue(s) of the constant domains bear a free thiol group. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical linkages of antibody fragments are also known.

[0058] 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 possible naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Each mAb is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they may be synthesized by hybridoma culture, free from contamination by other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies used in accordance with the present invention may be produced in immortalized B cells or hybridomas thereof, or may be produced by recombinant DNA methods.

[0059] Monoclonal antibodies herein include hybrid and recombinant antibodies produced by combining variable (including hypervariable) domains of a CD47 antibody with constant domains (e.g., "humanized" antibodies), or by combining light chains with heavy chains, or by combining chains from one species with chains from another species, or fusions with heterologous proteins, regardless of species of origin or immunoglobulin class or subclass designation, as well as antibody fragments (e.g., Fab, F(ab')2, and Fv), so long as they exhibit the desired biological activity.

[0060] Monoclonal antibodies herein specifically include chimeric antibodies (immunoglobulins) in which a portion of the heavy and / or light chain is identical or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity.

[0061] As used herein, the term "treatment" refers to a clinical intervention designed to change the natural history of the individual or cells being treated during the course of clinical pathology. Desirable treatment effects include a reduction in the rate of disease progression, amelioration or alleviation of disease symptoms, and remission or improved prognosis. For example, an individual is successfully "treated" if there is a reduction or elimination of one or more symptoms associated with cancer, including, but not limited to, a reduction (or destruction) in the proliferation of cancerous cells, a reduction in symptoms resulting from the disease, an improvement in the quality of life of the person suffering from the disease, a reduction in the dose of other drugs required to treat the disease, and / or an increase in the survival time of the individual. In some embodiments, "treating" a disease, such as cancer, refers to delaying the progression of the disease, i.e., prolonging, preventing, slowing, delaying, stabilizing, and / or postponing the onset of the disease (such as cancer). This delay may be of various lengths of time depending on the history of the disease and / or the individual undergoing the treatment. As will be apparent to one of skill in the art, a sufficient or significant delay may in fact encompass prevention, in that the individual does not develop the disease. For example, advanced stage cancer, such as the development of metastases, may be delayed.

[0062] An "effective amount" is at least the minimum amount required to bring about a measurable improvement or prevention of a particular disease (e.g., cancer). The effective amount herein may vary depending on factors such as the patient's condition, age, sex, and weight, and the ability of the therapeutic agent (or combination of therapeutic agents) to elicit a desired response in an individual. An effective amount is also an amount in which the therapeutically beneficial effects outweigh any toxic or adverse effects of the treatment. With respect to therapeutic use, beneficial or desired results include clinical results such as a reduction in one or more symptoms resulting from the disease, an improvement in the quality of life of the person suffering from the disease, a reduction in the dose of other drugs required to treat the disease, an enhancement of the effect of another drug, such as through targeting, a delay in the progression of the disease, and / or an increase in survival time. In the case of cancer or tumors, an effective amount of a drug may have an effect in reducing the number of cancer cells, reducing tumor size, inhibiting (i.e., slowing or desirably stopping) the invasion of cancer cells into peripheral organs, inhibiting (i.e., slowing or desirably stopping) tumor metastasis, inhibiting tumor growth to some extent, and / or alleviating to some extent one or more of the symptoms associated with the disorder. An effective amount may be administered in one or more administrations. For purposes of this disclosure, an effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to effect a therapeutic treatment, whether directly or indirectly. As will be understood in the clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administration of one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if a desired result can be or is achieved in conjunction with one or more other agents.

[0063] As used herein, the term "subject" for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and livestock animals, as well as zoo, sport, or pet animals, such as dogs, horses, cats, cows, etc. Preferably, the mammal is a human.

[0064] All references cited herein, including patent applications and publications, are hereby incorporated by reference in their entirety.

[0065] Overview This application is based on the unexpected finding that a combination treatment comprising a KRAS inhibitor and an agent that blocks the interaction between CD47 and SIRPα (e.g., an anti-CD47 antibody) is significantly more effective than either drug alone in inhibiting the growth of tumors containing cells expressing KRAS mutant proteins. Without being bound by theory, the synergistic anti-tumor effect of this combination is due, at least in part, to the fact that such a combination stimulates phagocytosis of tumor cells by macrophages to a greater extent than either drug alone. In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor (e.g., AMG 510), and the cancer or population of cancer cells (e.g., tumor) comprises one or more cells expressing KRAS G12C mutant proteins. In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor (e.g., AMG MRTX1133), and the cancer or population of cancer cells (e.g., tumor) comprises one or more cells expressing KRAS G12D mutant proteins.

[0066] Methods for treating cancer and stimulating phagocytosis of cancer cells by macrophages - Patents.com Provided herein are methods of treating cancer in a subject (i.e., a cancer comprising one or more cancer cells expressing a KRAS mutant protein (e.g., a KRAS G12C mutant protein or a KRAS G12D mutant protein)), the methods comprising administering to the subject an effective amount of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), and an effective amount of a KRAS inhibitor (e.g., a KRAS G12C inhibitor if the KRAS mutant protein is a KRAS G12C mutant protein, or a KRAS G12D inhibitor if the KRAS mutant protein is a KRAS G12D mutant protein). In some embodiments, the subject is a human. Also provided is a method of stimulating phagocytosis by macrophages of a population of cancer cells comprising one or more cancer cells expressing a KRAS mutant protein (e.g., a KRAS G12C mutant protein or a KRAS G12D mutant protein), the method comprising contacting the population with an effective amount of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and an effective amount of a KRAS inhibitor (e.g., a KRAS G12C inhibitor if the KRAS mutant protein is a KRAS G12C mutant protein, or a KRAS G12D inhibitor if the KRAS mutant protein is a KRAS G12D mutant protein). A cancer (or population of cancer cells) comprising one or more cancer cells expressing a KRAS mutant protein is alternatively referred to herein as a "KRAS-associated cancer" or a "KRAS mutant." In some embodiments, the KRAS mutant cancer is lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal cancer.

[0067] In some embodiments, the agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) is a polypeptide. In some embodiments, the agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) is a polypeptide that binds to CD47 (e.g., hCD47). In some embodiments, the polypeptide is or includes an anti-CD47 antibody, an immunologically active fragment thereof, or an antibody-based construct (such as a multispecific construct, e.g., a bispecific antibody). Exemplary anti-CD47 antibodies utilized with the methods are described in more detail below.

[0068] In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor, and the cancer (or population of cancer cells) comprises one or more cancer cells expressing a KRAS G12C mutant protein. Such cancers are alternatively referred to herein as "KRAS G12C-associated cancers" or "KRAS G12C mutant cancers." In some embodiments, the KRAS G12C mutant cancer is lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer is lung adenocarcinoma or non-small cell lung cancer (NSCLC). In some embodiments, the KRAS G12C inhibitor is, for example, a polypeptide (such as an antibody), peptide, antisense oligonucleotide, or small molecule that inhibits the activity of KRAS G12C mutant protein. In some embodiments, the KRAS G12C inhibitor is a small molecule. Exemplary small molecule KRAS G12C inhibitors that can be used with the methods provided herein include, but are not limited to, AMG 510 (Amgen / Beigene), MRTX849 (Mirati / Zai Lab), JAB-21822 (Jacobiopharma), GDC-6036 (Genentech), JDQ443 (Novartis), D-1553 (InventisBio and Merck Sharp & Dohme), GH35 (Genhouse Bio), GFH925 (GenFleet Therapeutics), BPI-421286 (Bettapharma), and LY3537982. Additional details regarding these and other exemplary small molecule KRAS G12C inhibitors are provided below.

[0069] In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor, and the cancer (or population of cancer cells) comprises one or more cancer cells expressing a KRAS G12D mutant protein. Such cancers are alternatively referred to herein as "KRAS G12D-associated cancers" or "KRAS G12D mutant cancers." In some embodiments, the KRAS G12D mutant cancer is lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12D mutant cancer is colon cancer, such as colorectal adenocarcinoma, or colon cancer, such as colon adenocarcinoma. In some embodiments, the KRAS G12D inhibitor is, for example, a polypeptide (such as an antibody), a peptide, an antisense oligonucleotide, or a small molecule that inhibits the activity of the KRAS G12D mutant protein. In some embodiments, the KRAS G12D inhibitor is a small molecule. Exemplary small molecule KRAS G12D inhibitors utilized with the methods provided herein include, but are not limited to, for example, MRTX1133. In some embodiments, the KRAS G12D inhibitor utilized with the methods provided herein is or includes an siRNA (also referred to herein as "KRAS G12D siRNA") that targets the mRNA encoding the KRAS G12D mutant protein. In some embodiments, the KRAS G12D siRNA is loaded into mesenchymal stromal cell-derived exosomes. Such exosomes are also known as "iExosomes" (see, e.g., Clinical Trial NCT03608631 and Surana et al. (2022) Journal of Clinical Oncology, 40:4_suppl, TPS633-TPS633). Additional details regarding these KRAS G12D inhibitors are provided below.

[0070] In some embodiments, the agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and the KRAS inhibitor are administered simultaneously. In some embodiments, "co-administration" means that the agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and the KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered about 15 minute(s) or less apart, such as about 10 minutes or less, about 5 minutes or less, or about 1 minute or less apart. In some embodiments, the simultaneous administration of an agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and a KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) can be combined with additional doses of an agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and / or a KRAS inhibitor. In some embodiments, the agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and a KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered sequentially. In some embodiments, "sequential administration" means that the agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and the KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered more than about 15 minutes apart, e.g., more than about 20 minutes, more than about 30 minutes, more than about 40 minutes, more than about 50 minutes, more than about 60 minutes, or more. For example, in some embodiments, the agent that blocks the interaction between CD47 and SIRPα is administered before the small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor). In some embodiments, the small molecule KRAS inhibitor is administered before the agent that blocks the interaction between CD47 and SIRPα.In some embodiments, the administration of the agent that blocks the interaction between CD47 and SIRPα and the KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) is concurrent, i.e., the administration periods of the agent that blocks the interaction between CD47 and SIRPα and the KRAS inhibitor overlap with each other. In some embodiments, the administration of the agent that blocks the interaction between CD47 and SIRPα and the KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) is non-concurrent.

[0071] Exemplary Anti-CD47 Antibodies An anti-CD47 antibody (or an immunologically active fragment thereof) is an antibody that binds to CD47 (e.g., human CD47 or "hCD47") with sufficient affinity and specificity. As used herein, an "immunologically active fragment" of an antibody refers to an antigen-binding fragment of an antibody. The terms "immunologically active fragment" and "antigen-binding fragment" are used interchangeably herein. In some embodiments, an anti-CD47 antibody (or an immunologically active fragment thereof) is a chimeric (e.g., humanized) monoclonal antibody (or an immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is selected from the group consisting of 5F9 (also known as Hu5F9-G4 and magrolimab), being developed by Gilead Sciences / Forty Seven, Inc., CC-90002 (also known as INBRX103), being developed by Celgene, LQ001, being developed by Novamab, HLX24, being developed by Henlius, TI-061, being developed by Arch Oncology (formerly known as Tioma Therapeutics), AO-176, being developed by Arch Oncology, SRF-231, being developed by Surface Oncology, IBI-188, being developed by Innovent Bio, AK117, being developed by Akesobio, IMC-002, being developed by ImmuneOncia Therapeutics 3D Medicines, SHR-1603, being developed by Jiangsu HengRui Medicine, Sorrento Therapeutics In some embodiments, the immunologically active fragment of the anti-CD47 antibody is an immunologically active fragment of any one of the aforementioned anti-CD47 antibodies. Additional details about these exemplary anti-CD47 antibodies can be found, for example, in Jiang et al. (2021) J Hematol Oncol 14:180 doi(dot)org / 10(dot)1186 / s13045-021-01197-w, WO2011 / 143624A2, USP9,382,320B2.

[0072] In some embodiments, the anti-CD47 antibody (or immunologically active fragment thereof) comprises a heavy chain variable domain (V H ), and / or a light chain variable domain (V L ).

[0073] In some embodiments, an anti-CD47 antibody (or an immunologically active fragment thereof) comprises: (a) a VH domain comprising (1) a CDR-H1 comprising RAWMN (SEQ ID NO:5), (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO:6), (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO:7), and (b) a VL domain comprising (1) a CDR-L1 comprising KSSQSVLYAGNNRNYLA (SEQ ID NO:8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO:9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO:10). In some embodiments, the CDR sequences are defined according to Kabat (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). In some embodiments, an anti-CD47 antibody (or an immunologically active fragment thereof) comprises: (a) a VH domain comprising (1) a CDR-H1 comprising GLTFERA (SEQ ID NO: 11), (2) a CDR-H2 comprising KRKTDGET (SEQ ID NO: 12), (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 7), and (b) a VL domain comprising (1) a CDR-L1 comprising KSSQSVLYAGNNRNYLA (SEQ ID NO: 14), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 15), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 16). In some embodiments, the CDR sequences are defined according to the Chothia numbering system (see, e.g., Chothia and Lesk (1986) EMBO J. 5(4):823-6 and Al-Lazikani et al., (1997) JMB 273:927-948).In some embodiments, the anti-CD47 antibody (or immunologically active fragment thereof) comprises: (a) a VH domain comprising (1) a CDR-H1 comprising GLTFERAW (SEQ ID NO: 17); (2) a CDR-H2 comprising IKRKTDGETT (SEQ ID NO: 18); (3) a CDR-H3 comprising AGSNRAFDI (SEQ ID NO: 19); and (b) a VL domain comprising (1) a CDR-L1 comprising QSVLYAGNNRNY (SEQ ID NO: 20); (2) a CDR-L2 comprising QAS (SEQ ID NO: 21); and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 22). In some embodiments, the CDR sequences are defined according to the IMGT numbering system (see, e.g., Lefranc MP. (2013) IMGT Unique Numbering. In: Dubitzky W., Wolkenhauer O., Cho KH., Yokota H. (eds) Encyclopedia of Systems Biology. Springer, New York, NY, https: / / doi.org / 10.1007 / 978-1-4419-9863-7_127). In some embodiments, the anti-CD47 antibody (or immunologically active fragment thereof) comprises: (a) a VH domain comprising (1) a CDR-H1 comprising GLTFERAWMN (SEQ ID NO: 23); (2) a CDR-H2 comprising RIKRKTDGETTD (SEQ ID NO: 24); (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 25); and (b) a VL domain comprising (1) a CDR-L1 comprising KSSQSVLYAGNNRNYLA (SEQ ID NO: 26); (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 27); and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 28). In some embodiments, the CDR sequences are defined according to the AbM numbering system (see, e.g., Abhinandan RK, Martin ACA analysis and improvements to Kabat and structurally correct numbering of antibody variable domains. Mol. Immunol. 2008; 45: 3832-3839. doi: 10.1016 / j.molimm.2008.05.022).In some embodiments, an anti-CD47 antibody (or an immunologically active fragment thereof) comprises: (a) a VH domain comprising (1) a CDR-H1 comprising ERAWMN (SEQ ID NO:29), (2) a CDR-H2 comprising WVGRIKRKTDGETTD (SEQ ID NO:30), (3) a CDR-H3 comprising AGSNRAFD (SEQ ID NO:31), and (b) a VL domain comprising (1) a CDR-L1 comprising LYAGNNRNYLAWY (SEQ ID NO:32), (2) a CDR-L2 comprising LLINQASTRA (SEQ ID NO:33), and (3) a CDR-L3 comprising QQYYTPPL (SEQ ID NO:34). In some embodiments, the CDR sequences are defined according to the Contact numbering system (see, e.g., McCallum et al. (1996) J Mol Biol. 262(5):732-45, doi:10.1006 / jmbi.1996.0548).

[0074] For ease of reference, the amino acid sequences of SEQ ID NOs:5-34 are provided in Table A below. [Table 4-1] [Table 4-2]

[0075] In some embodiments, the anti-CD47 antibody (or immunologically active fragment thereof) comprises the three CDRs of a VH domain comprising SEQ ID NO:1. Additionally or alternatively, in some embodiments, the anti-CD47 antibody (or immunologically active fragment thereof) comprises the three CDRs of a VL domain comprising SEQ ID NO:2. In some embodiments, the three CDRs of the VH domain are CDRs according to the Kabat, Chothia, AbM, or Contact numbering system. Additionally or alternatively, in some embodiments, the three CDRs of the VL domain are CDRs according to the Kabat, Chothia, AbM, or Contact numbering system. In some embodiments, the N-terminal amino acid of the VH domain of the anti-CD47 antibody (or immunologically active fragment thereof) is E, and optionally, in some embodiments, the C-terminal amino acid of the VH domain of the anti-CD47 antibody (or immunologically active fragment thereof) is S.

[0076] In some embodiments, the anti-CD47 antibody (or immunologically active fragment thereof) comprises a heavy chain variable domain (VH) comprising an amino acid sequence having at least about 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO:1. In some embodiments, the N-terminal amino acid of the VH domain having at least about 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 is E. Additionally or alternatively, in some embodiments, the C-terminal amino acid of the VH domain having at least about 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 is S. In some embodiments, the VH domain of the anti-CD47 antibody (or immunologically active fragment thereof) comprises a heavy chain variable domain (VH) comprising an amino acid sequence having at least about 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 is E. Additionally or alternatively, in some embodiments, the C-terminal amino acid of the VH domain having at least about 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 is S. H The N-terminal amino acid of the domain corresponds to position H1 according to the Kabat numbering system and corresponds to position V of an anti-CD47 antibody (or an immunologically active fragment thereof). H The C-terminal amino acid of the domain corresponds to position H113 according to the Kabat numbering system. HThe N-terminal amino acid of the domain corresponds to position H1 according to the Chothia numbering system and corresponds to position V of an anti-CD47 antibody (or an immunologically active fragment thereof). H The C-terminal amino acid of the domain corresponds to position H113 according to the Chothia numbering system. H The N-terminal amino acid of the domain corresponds to position H1 according to the IMGT numbering system and corresponds to position V of an anti-CD47 antibody (or an immunologically active fragment thereof). H The C-terminal amino acid of the domain corresponds to position H128 according to the IMGT numbering system. H The N-terminal amino acid of the domain corresponds to amino acid 1 of SEQ ID NO:1, H The C-terminal amino acid of the domain of an anti-CD47 antibody (or an immunologically active fragment thereof) corresponds to amino acid 118 of SEQ ID NO: 1. In some embodiments, an anti-CD47 antibody comprises (e.g., further comprises) a light chain variable domain (VL) comprising an amino acid sequence having at least about 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, an anti-CD47 antibody comprises a VH comprising SEQ ID NO: 1 and a VL comprising SEQ ID NO: 2. The amino acid sequences of SEQ ID NOs: 1 and 2 are provided below. [ka]

[0077] In some embodiments, the anti-CD47 antibody is a full-length antibody. In some embodiments, the full-length antibody comprises a human Fc region. In some embodiments, the human Fc region is an IgG1, IgG2, or IgG4 Fc region. In some embodiments, the full-length anti-CD47 antibody comprises a human IgG4 Fc region comprising a S228P substitution, or a variant thereof, where the amino acid numbering is according to the EU index. In some embodiments, the full-length anti-CD47 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:3 and a light chain comprising the amino acid sequence of SEQ ID NO:4. In some embodiments, the full-length anti-CD47 antibody is a full-length antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:35 and a light chain comprising the amino acid sequence of SEQ ID NO:4. In some embodiments, the anti-CD47 antibody is lemzoparlimab (also known as TJ011133). [ka]

[0078] An anti-CD47 antibody that specifically binds to hCD47 can be of any of the various types of antibodies as defined above, but in certain embodiments is a human, humanized, or chimeric antibody. In some embodiments, the anti-CD47 antibody is a human antibody. In some embodiments, the anti-CD47 is a humanized antibody that includes a constant domain of a human antibody (e.g., a human Fc domain, e.g., a human IgG Fc domain, e.g., a human IgG1, human IgG2, human IgG3, or human IgG4 Fc domain, or a variant of a human IgG4 Fc domain, including a S228P substitution (wherein amino acid numbering is according to the EU index). In some embodiments, the anti-CD47 antibody is a chimeric antibody. See, e.g., U.S. Pat. No. 4,816,567 and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984). In some embodiments, a chimeric anti-CD47 antibody comprises a non-human variable region (e.g., a variable region derived from a non-human primate such as chicken, mouse, rat, hamster, rabbit, or monkey) and a human constant region. In some embodiments, a chimeric antibody is a "class-switched" antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0079] In some embodiments, a chimeric antibody is a humanized antibody. A non-human antibody may be humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the non-human parent antibody. Generally, a humanized antibody comprises one or more variable domains in which the HVR, e.g., CDR (or a portion thereof) is derived from a non-human antibody (e.g., a chicken antibody) and the FR (or a portion thereof) is derived from a human antibody sequence. A humanized antibody also optionally comprises at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are replaced with the corresponding residues from a non-human antibody (e.g., the antibody from which the HVR or CDR residues are derived), e.g., to restore or improve antibody specificity or affinity. Humanized antibodies and methods for their production are reviewed, e.g., in Almagro and Fransson, Front.Biosci.13:1619-1633 (2008).

[0080] Human framework regions useful for humanization include, but are not limited to, framework regions selected using the "best-fit" method; framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions; somatically mutated human framework regions or human germline framework regions; and framework regions derived from screening of FR libraries.See, for example, Sims et al. J.Immunol. 151:2296 (1993); Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al. J.Immunol., 151:2623 (1993); Almagro and Fransson, Front.Biosci. 13:1619-1633 (2008); and Baca et al., J.Biol.Chem. 272:10678-10684 (1997).

[0081] In some embodiments, the anti-CD47 antibodies of the present disclosure are human antibodies. Human antibodies can be produced using a variety of techniques known in the art. In some embodiments, human antibodies are produced by genetically engineered non-human animals, such as chickens (see, e.g., U.S. Pat. Nos. 8,592,644 and 9,380,769) and / or mice as described herein. Human antibodies are generally described in Lonberg, Curr. Opin. Immunol. 20:450-459 (2008).

[0082] In some embodiments, the anti-CD47 antibodies of the present disclosure are antibody fragments (e.g., immunologically active fragments), including, but not limited to, Fab, F(ab')2, Fab'-SH, Fv, or scFv fragments, or single domain, single heavy chain, or single light chain antibodies. As used herein, an "immunologically active fragment" of an antibody refers to an antigen-binding fragment of an antibody. The terms "immunologically active fragment" and "antigen-binding fragment" are used interchangeably herein. Antibody fragments can be generated, for example, by enzymatic digestion or by recombinant techniques. In some embodiments, antibody fragments are generated using proteolytic digestion of intact antibodies, for example, as described in Morimoto et al., Journal of Biochemical and Biophysical Methods 24:107-117 (1992) and Brennan et al., Science, 229:81 (1985). In some embodiments, antibody fragments are produced by recombinant host cells. For example, Fab, Fv and ScFv antibody fragments can be expressed by and secreted from E. coli. Antibody fragments can alternatively be isolated from antibody phage libraries. Other methods for producing immunologically active fragments of antibodies are known in the art.

[0083] In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) specifically recognizes (e.g., binds to) hCD47 expressed on the surface of a cell. In some embodiments, the anti-CD47 antibody specifically recognizes hCD47 expressed on the surface of a cancer cell, e.g., a KRAS G12C mutant cancer cell or a KRAS G12D mutant cancer cell (such as a lung adenocarcinoma cell, a non-small cell lung cancer (NSCLC) cell, a colon adenocarcinoma cell, a colorectal adenocarcinoma cell, a pancreatic cancer cell, a cholangiocarcinoma cell, an endometrial cancer cell, an ovarian cancer cell, a peritoneal cancer cell, a bladder cancer cell, a gastric cancer cell, a thyroid cancer cell, a melanoma cell, a breast cancer cell, a head and neck cancer cell, a multiple myeloma cell, an acute myeloid leukemia (AML) cell, a uterine cancer cell, a gastric or esophageal cancer cell, or a rectal cancer cell). In some embodiments, binding of an anti-CD47 antibody described herein (or an immunologically active fragment thereof) to hCD47 (e.g., hCD47 expressed on the surface of a cell) blocks the interaction of hCD47 with signal regulatory protein alpha (SIRPα), e.g., human SIRPα ("hSIRPα"). In some embodiments, binding of an anti-CD47 antibody described herein (or an immunologically active fragment thereof) to hCD47 expressed on the surface of a cancer cell (e.g., a KRAS G12C mutant cancer cell or a KRAS G12D mutant cancer cell) promotes macrophage-mediated phagocytosis of the cancer cell.

[0084] Further details regarding exemplary anti-CD47 antibodies (including pharmaceutical formulations containing same, exemplary dosages, and exemplary administration schedules) that may be used with methods of treating KRAS mutant cancers (e.g., KRAS G12C mutant cancers or KRAS G12D mutant cancers) are provided in PCT / CN2021 / 123892. Further details regarding methods of producing / manufacturing the anti-CD47 antibodies described herein are provided in PCT / CN2021 / 123892.

[0085] KRAS and Exemplary KRAS Inhibitors KRAS, a member of the RAS family, is a key regulator of signaling pathways responsible for cell proliferation, differentiation, and survival. See, e.g., Cox et al. (2003) Nat Rev Drug Discov. 13(11):828-851 and Downward J. (2003) Nat Rev Cancer. 3(1):11-22. KRAS is the most frequently mutated oncogene in human cancers, and mutations in KRAS can lead to continued cell proliferation and cancer development. KRAS G12C mutations are found in approximately 13% of NSCLC patients and 1%-3% of colorectal and other solid tumors. G12C is a single point mutation with a glycine to cysteine ​​substitution at codon 12. See Cox et al. (2003) Nat Rev Drug Discov. 13(11):828-851, Neumann et al. (2009) Pathol Res Pract. 205:858-862, and Biernacka et al. (2016) Cancer Genet. 209(5):195-198. This substitution favors an activated, GTP-bound state of KRAS, amplifying the signaling pathways that lead to carcinogenesis (see Ryan et al. (2018) Nat Rev Clin Oncol. 15(11):709-720 and Kim et al. (2020 Cell, 183(4):850-859). KRAS G12D mutations are found in approximately one-third of pancreatic cancers and one-tenth of colorectal cancers. The glycine-to-aspartic acid mutation at position 12 (G12D) leads to a bulkier, negatively charged side chain protruding into the active site, which causes steric hindrance in GTP hydrolysis (see Malumbres et al. (2003) Nat Rev Cancer, 3,459-465), impairing GTPase function and locking KRAS into its active (GTP-bound) state (Scheffzek et al. (1997) Science 277, 333-338).

[0086] In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor, and the cancer (or population of cancer cells) comprises one or more cells expressing a KRAS G12C mutant protein. In some embodiments, the KRAS G12C mutant cancer is lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer is lung adenocarcinoma or non-small cell lung cancer (NSCLC).

[0087] In some embodiments, the term "KRAS G12C inhibitor" refers to any agent that inhibits the activity of drug KRAS G12C mutant protein, such as polypeptide, fusion polypeptide, antibody, peptide, antisense oligonucleotide, or small molecule.In some embodiments, KRAS G12C inhibitor directly interacts with KRAS G12C mutant protein and inhibits the activity of said protein.In some embodiments, KRAS G12C inhibitor is a small molecule drug. Exemplary small molecule KRAS G12C inhibitors for use with the methods provided herein include, but are not limited to, AMG 510 (also known as sotorasib, LUMAKRAS™, and LUMYKRAS™), MRTX849 (also known as adagrasib), JAB-21822 (also known as JAB-21000), GDC-6036, JDQ443, D-1553, GH35, GFH925, BPI-421286, or LY3537982. Other exemplary small molecule KRAS G12C inhibitors are described, for example, in Hillig et al. (2019) Proc Natl Acad Sci US A. 116(7):2551-2560 and Sun et al. (2012) Angew Chem Int Ed. Engl. 51(25):6140-6143, as well as WO2020 / 233592, WO2021 / 023247, WO2021 / 083167, WO2020 / 238791, WO2021 / 000885, CN112585129, CN112552295, CN112390818, CN112390796, WO2019 / 141250, CN111592528, WO2020 / 259432, CN112300153, CN112552294, WO2020 / 239123, CN11 0698378, CN111377918, CN111205286, CN112047939, WO2020 / 259513, WO2021 / 023154, CN112430234, WO2021 / 063346, WO2021 / 058018, C N112225734, WO2021 / 155716, CN112159405, CN112778302, CN112830928, WO2020 / 1027943, CN112574199, WO2021 / 037018, CN110172089,WO2020 / 156285, CN111499634, WO2020 / 177629, WO2020 / 216190, WO2020 / 221239, WO2020 / 233592, WO2020 / 238791, WO2020 / 239077, WO2020 / 239123 , CN112047933, CN112047937, CN112047948, WO2020 / 259513, WO2020 / 259573, WO2020 / 259432, WO2021 / 000885, CN112174950, CN112300153, WO2021 / 023154, WO2021 / 023247, CN112390818, WO2021 / 027943, WO2021 / 027911, WO2021 / 031952, CN112390796, WO2021 / 037018, CN112442029, WO2021 / 0433 22, WO2021 / 052499, CN112538084, CN112552295, WO2021 / 058018, WO2021 / 063346, WO2021 / 068898, WO2021 / 078285, CN112225734, CN112707905, CN1 12745335, WO2021 / 083167, CN112778284, WO2021 / 088458, CN112851663, WO2021 / 093758, WO2021 / 098859, CN112830928, CN111377918, WO2021 / 1044 31, WO2021 / 109737, WO2021 / 113595, CN112920183, WO2021 / 121371, WO2021 / 121367, CN113004269, WO2021 / 129824, WO2021 / 129820, CN113061132, W WO2021 / 139678, WO2021 / 139748, CN111205286, WO2021 / 143693, CN113135924, WO2021 / 147965, WO2021 / 155716, WO2021 / 168193, WO2021 / 169990, WO2021 / 169963, CN113321654, WO2021 / 175199, WO2021 / 180181, WO2021 / 185233, WO2021 / 190467, WO2021 / 197499, CN112574199, and CN112300269,The contents of this document are incorporated herein by reference in their entirety.

[0088] In some embodiments, the KRAS G12C inhibitor is AMG 510, also known as sotorasibe, LUMAKRAS™, and LUMYKRAS™, as described above. AMG 510 is currently in development by Amgen / Beigene. AMG 510 can exist in either of two atropisomers, one of which is more active than the other (see, for example, https: / / cen(dot)acs(dot)org / pharmaceuticals / drug(dash)discovery / Amgen(dash)unveils(dash)KRas(dash)inhibitor(dash)human / 97 / i14). AMG 510 selectively forms an irreversible covalent bond to the sulfur atom in a cysteine ​​residue that is present in the G12C mutant form of the KRAS protein but not in the wild-type form. Covalent binding of AMG 150 to KRAS G12C locks the protein in its inactive GDP-bound conformation, thus inhibiting KRAS-dependent signaling. AMG 150 has the empirical formula C 30 HF2N6O3 and has a molecular weight of 560.606 g / mol. AMG 150 is chemically described as 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one and has the following chemical structure: [ka] The CAS registration number for AMG 510 is 2252403-56-6. The efficacy of AMG 510 has been demonstrated in a subset of patients enrolled in a single-arm, open-label, multicenter study (NCT03600883) and is currently being investigated in further clinical trials. Complete information on the preparation, formulation, dosage, and administration schedule of AMG 510 can be found in the local package insert (in the United States, see, for example, www(dot)accessdata(dot)fda(dot)gov / drugsatfda_docs / label / 2021 / 214665s000lbl.pdf). Further details regarding the structure and synthesis of AMG 510 are provided in WO2018 / 217651, the contents of which are incorporated herein by reference in their entirety.

[0089] In some embodiments, the KRAS G12C inhibitor is MRTX849 (also known as adagrasib). MRTX849 is currently in development by Mirati / Zai Lab. MRTX849 selectively forms an irreversible covalent bond to the sulfur atom in a cysteine ​​residue that is present in the G12C mutant form of KRAS protein but not in the wild-type form. The covalent binding of MRTX849 to KRAS G12C locks the protein in its inactive GDP-bound conformation, thus inhibiting KRAS-dependent signaling. MRTX849 has the empirical formula C 32 H 35 It has ClFN7O2 and a molecular weight of 604.13 g / mol. MRTX849 is chemically described as 2-[(2S)-4-[7-(8-chloronaphthalen-1-yl)-2-[[(2S)-1-methylpyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-1-(2-fluoroprop-2-enoyl)piperazin-2-yl]acetonitrile and has the following chemical structure: [ka] The CAS Registry Number for MRTX849 is 2326521-71-3. MRTX849 is currently being evaluated in several clinical trials, including NCT04613596, NCT04685135, NCT03785249, NCT04330664, etc. Further details regarding the structure and synthesis of MRTX849 are provided in Fell et al. (2020) J. Med. Chem. 63, 6679-6693 and WO2017 / 201161, the contents of which are incorporated herein by reference in their entirety.

[0090] In some embodiments, the KRAS G12C inhibitor is JAB-21822. JAB-21822 is being developed by Jacobio Pharmaceuticals Group Co., LTD. (see, for example, en(dot)jacobiopharma(dot)com / blank3(dot)html?introId=23 and www(dot)jacobiopharma(dot)com / news / 207(dot)html). JAB-21822 is currently being evaluated in several clinical trials, including NCT05009329 and NCT05002270. Further details regarding the structure and synthesis of JAB-21822 are provided in WO2021 / 057832, the contents of which are incorporated herein by reference in their entirety.

[0091] In some embodiments, the KRAS G12C inhibitor is GDC-6036. GDC-6036 is being developed by Genentech, Inc. (see, e.g., www(dot)genentechoncology(dot)com / pipeline-molecules / kras-g12c.html) and is currently being evaluated in clinical trial NCT04449874. Further details regarding the structure and synthesis of GDC-6036 are provided in WO2020 / 097537, the contents of which are incorporated herein by reference in their entirety.

[0092] In some embodiments, the KRAS G12C inhibitor is JDQ443. JDQ443 is being developed by Novartis. The structure of JDQ443 is: [ka] JDQ443 is currently being evaluated in clinical trial NCT04699188. Further details regarding the synthesis of JDQ443 are provided in WO2021 / 124222, the contents of which are incorporated herein by reference in their entirety.

[0093] In some embodiments, the KRAS G12C inhibitor is D-1553. D-1553 is being developed by InventisBio Co., Ltd. (see, e.g., www(dot)inventisbio(dot)com / %e4%b8%b4%e5%ba%8a%e8%af%95%e9%aa%8c / ) and is currently being evaluated in clinical trial NCT04585035 in collaboration with Merck Sharp & Dohme. Further details regarding the structure and synthesis of D-1553 are provided in WO2020 / 233592, the contents of which are incorporated herein by reference in their entirety.

[0094] In some embodiments, the KRAS G12C inhibitor is GH35. GH35 is being developed by Suzhou Genhouse Bio Co., Ltd. (see, for example, www(dot)genhousebio(dot)com / en / product / index(dot)html) and is being evaluated in clinical trial NCT05010694. Further details regarding the structure and synthesis of GH35 are provided in WO2020 / 177653, the contents of which are incorporated herein by reference in their entirety.

[0095] In some embodiments, the KRAS G12C inhibitor is GFH925. GFH925 is being developed by GenFleet Therapeutics (Zhejiang) (see, e.g., www(dot)genfleet(dot)com / en / science) and is being evaluated in clinical trial NCT05005234. Further details regarding the structure and synthesis of GFH925 are provided in WO2020 / 177629, WO2020 / 221239, and WO2021 / 031952, the contents of which are incorporated herein by reference in their entirety.

[0096] In some embodiments, the KRAS G12C inhibitor is BPI-421286. BPI-421286 is being developed by Betta Pharmaceutical Co.,Ltd (see, for example, www(dot)bettapharma(dot)com / News / show / id / 2380), and clinical trial applications (CXHL2100046 and CXHL2100047) for the evaluation of BPI-421286 have been approved by the State Food and Drug Administration of the People's Republic of China. Further details regarding the structure and synthesis of BPI-421286 are provided in CN112390796, the contents of which are incorporated herein by reference in their entirety.

[0097] In some embodiments, the KRAS G12C inhibitor is LY3537982. LY3537982 is being developed by Eli Lilly and Company and Loxo Oncology, Inc. (see, e.g., www(dot)lillyloxooncologypipeline(dot)com / molecule / kras-g12c-inhibitor / ) and is being investigated in clinical trial NCT04956640. Further details regarding the structure and synthesis of LY3537982 are provided in WO2021 / 118877, the contents of which are incorporated herein by reference in their entirety.

[0098] In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor, and the cancer (or population of cancer cells) comprises one or more cells expressing a KRAS G12D mutant protein. In some embodiments, the KRAS G12D mutant cancer is lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12D mutant cancer is a colon cancer, such as a colorectal adenocarcinoma, or a colon cancer, such as a colon adenocarcinoma. In some embodiments, the term "KRAS G12D inhibitor" refers to any agent, e.g., a polypeptide, a fusion polypeptide, an antibody, a peptide, an antisense oligonucleotide, or a small molecule, that inhibits the activity of a drug KRAS G12D mutant protein. In some embodiments, the KRAS G12D inhibitor directly interacts with the KRAS G12D mutant protein to inhibit the activity of the protein. In some embodiments, the KRAS G12D inhibitor is a small molecule drug. An exemplary small molecule KRAS G12D inhibitor utilized with the methods provided herein is MRTX1133. In some embodiments, the KRAS G12D inhibitor utilized with the methods provided herein is or includes an siRNA (also referred to herein as "KRAS G12D siRNA") that targets the mRNA encoding the KRAS G12D mutant protein. In some embodiments, the KRAS G12D siRNA is loaded into mesenchymal stromal cell-derived exosomes, also known as "iExosomes" (see, e.g., Clinical Trial NCT03608631 and Surana et al. (2022) Journal of Clinical Oncology, 40:4_suppl, TPS633-TPS633).

[0099] In some embodiments, the KRAS G12D inhibitor is MRTX1133. MRTX1133, which is being developed by Mirati Therapeutics, Inc., exhibits selective and reversible inhibition of both active and inactive states of KRAS G12D mutant proteins (Wang et al. (2022) J Med Chem, 65(4):3123-3133). The specificity of MRTX1133 for KRAS G12D mutant proteins is more than 1000-fold greater than that of wild-type KRAS (Wang et al. (2022) J Med Chem, 65(4):3123-3133). MRTX1133 has the empirical formula C 33 H 31 F3N6O2 and has a molecular weight of 600.63 g / mol. MRTX1133 has the following chemical structure: [ka] The CAS Registry Number for MRTX1133 is 2621928-55-8. MRTX1133 is currently being evaluated in clinical trial NCT05737706. Further details regarding MRTX1133 are provided, for example, in Wang et al. (2022) J Med Chem, 65(4):3123-313, Xie et al. (2021) Front Oncol, 11:672612, and Hallin et al. (2022) Nat Med, 28(10):2171-2182.

[0100] In some embodiments, the KRAS G12D inhibitor is or includes an siRNA (also referred to herein as "KRAS G12D siRNA") targeting an mRNA encoding a KRAS G12D mutant protein. In some embodiments, the KRAS G12D siRNA is loaded into bone marrow mesenchymal / stromal cell-derived exosomes. Such exosomes are known as "iExosomes" (see, e.g., Clinical Trial NCT03608631 and Surana et al. (2022) Journal of Clinical Oncology, 40:4_suppl, TPS633-TPS633). Additional details regarding iExosomes are provided in Mendt et al. (2018) JCI Insight, 3(8):e99263 and Kamerkar et al. (2017) Nature. 546:498-503.

[0101] Exemplary Methods In some embodiments, a method of treating a KRAS mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS mutant protein, e.g., a KRAS G12C mutant protein or a KRAS G12D mutant protein) in a subject (e.g., a human subject) is provided, the method comprising administering to the subject an effective amount of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), and an effective amount of a KRAS inhibitor (e.g., a KRAS G12C inhibitor if the cancer is a KRAS G12C mutant cancer, or a KRAS G12D inhibitor if the cancer is a KRAS G12C mutant cancer). In some embodiments, a method is provided for stimulating phagocytosis of a population of cancer cells comprising one or more cancer cells expressing a KRAS mutant protein (e.g., a KRAS G12C mutant protein or a KRAS G12D mutant protein), the method comprising contacting the population with an effective amount of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and an effective amount of a KRAS inhibitor (e.g., a KRAS G12C inhibitor if the KRAS mutant protein is a KRAS G12C mutant protein, or a KRAS G12D inhibitor if the KRAS mutant protein is a KRAS G12D mutant protein). In some embodiments, the KRAS mutant cancer (e.g., KRAS G12C mutant cancer or KRAS G12D mutant cancer), or the one or more cancer cells expressing a KRAS mutant protein (e.g., KRAS G12C mutant protein or KRAS G12D mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS mutant cancer is a KRAS G12C mutant cancer. In some embodiments, the population of cancer cells comprises one or more cells expressing a KRAS G12C mutant protein.In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing KRAS G12C mutant protein) is NSCLC. In some embodiments, the KRAS mutant cancer is a KRAS G12D mutant cancer. In some embodiments, the population of cancer cells comprises one or more cells expressing KRAS G12D mutant protein. In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing KRAS G12D mutant protein) is a colorectal cancer, e.g., colorectal adenocarcinoma, or a colon cancer, e.g., colon adenocarcinoma. In some embodiments of the method of treatment, the agent that blocks the interaction between CD47 and SIRPα and the KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered simultaneously. In some embodiments of the method of treatment, the agent that blocks the interaction between CD47 and SIRPα and the KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered sequentially. In some embodiments of the method of treatment, the agent that blocks the interaction between CD47 and SIRPα is administered before the KRAS inhibitor. In some embodiments of the method of treatment, the KRAS inhibitor is administered before the agent that blocks the interaction between CD47 and SIRPα. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is a polypeptide. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody or antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, shown below. [ka] In some embodiments, the KRAS G12C inhibitor is MRTX849, shown below. [ka]

[0102] In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor. In some embodiments, the KRAS G12D inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12D inhibitor is MRTX1122, as shown below. [ka]

[0103] In some embodiments, a method of treating a KRAS mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS mutant protein, e.g., a KRAS G12C mutant protein or a KRAS G12D mutant protein) in a subject (e.g., a human subject) is provided, the method comprising administering to the subject an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), and an effective amount of a KRAS G inhibitor (e.g., a KRAS G12C inhibitor if the cancer is a KRAS G12C mutant cancer, or a KRAS G12D inhibitor if the cancer is a KRAS G12C mutant cancer). In some embodiments, a method is provided for stimulating phagocytosis of a population of cancer cells comprising one or more cancer cells expressing a KRAS G12C mutant protein, the method comprising contacting the population with an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), and an effective amount of a KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing a KRAS G12C mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing a KRAS G12C mutant protein) is NSCLC. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the KRAS G12C inhibitor.In some embodiments of the method of treatment, the KRAS G12C inhibitor is administered prior to the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, or ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the anti-CD47 antibody (or immunologically active fragment thereof) is an anti-CD47 antibody (or immunologically active fragment thereof) described herein. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above.

[0104] In some embodiments, a method of treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject) is provided, comprising administering to the subject an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) and an effective amount of a KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer is NSCLC. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the KRAS G12C inhibitor. In some embodiments, the KRAS G12C inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above.

[0105] In some embodiments, a method of treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject) is provided, the method comprising administering to the subject (a) an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising: (1) a VH domain comprising: (1) a CDR-H1 comprising: (2) a CDR-H2 comprising: (1) RAWMN (SEQ ID NO:5); (2) a CDR-H2 comprising: (3) a CDR-H3 comprising: SNRAFDI (SEQ ID NO:7); and (b) a VL domain comprising: (1) a CDR-L1 comprising: KSSQSVLYAGNNRNYLA (SEQ ID NO:8); (2) a CDR-L2 comprising: QASTRAS (SEQ ID NO:9); and (3) a CDR-L3 comprising: QQYYTPPLA (SEQ ID NO:10); and an effective amount of a small molecule KRAS G12C inhibitor. In some embodiments, a method of stimulating phagocytosis of a population of cancer cells comprising one or more cancer cells expressing a KRAS G12C mutant protein is provided, the method comprising contacting the population with (a) an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a VH domain comprising: (1) a CDR-H1 comprising RAWMN (SEQ ID NO:5); (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO:6); (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO:7); and (b) a VL domain comprising: (1) a CDR-L1 comprising KSSQSVLYAGNNRNYLA (SEQ ID NO:8); (2) a CDR-L2 comprising QASTRAS (SEQ ID NO:9); and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO:10); and an effective amount of a small molecule KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing a KRAS G12C mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal cancer.In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing a KRAS G12C mutant protein) is NSCLC. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12C inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12C inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above.

[0106] In some embodiments, a method of treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject) is provided, the method comprising administering to the subject (a) an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising: (1) a VH domain comprising: (1) a CDR-H1 comprising: (2) a CDR-H2 comprising: (1) RAWMN (SEQ ID NO:5); (2) a CDR-H2 comprising: (3) a CDR-H3 comprising: SNRAFDI (SEQ ID NO:7); and (b) a VL domain comprising: (1) a CDR-L1 comprising: KSSQSVLYAGNNRNYLA (SEQ ID NO:8); (2) a CDR-L2 comprising: QASTRAS (SEQ ID NO:9); and (3) a CDR-L3 comprising: QQYYTPPLA (SEQ ID NO:10); and an effective amount of a small molecule KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer or the KRAS G12D mutant cancer is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS mutant cancer is a KRAS G12C mutant cancer. In some embodiments, the KRAS G12C mutant cancer is NSCLC. In some embodiments, the KRAS mutant cancer is a KRAS G12D mutant cancer. In some embodiments, the KRAS G12D mutant cancer is a colon cancer, e.g., colorectal adenocarcinoma, or a colon cancer, e.g., colon adenocarcinoma. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered prior to the small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor).In some embodiments of the treatment method, a small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) is administered prior to the anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above. In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor. In some embodiments, the KRAS G12D inhibitor is MTRX1133, the structure of which is shown above.

[0107] In some embodiments, a method is provided for treating a KRAS mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS G12C mutant protein, e.g., a KRAS G12C mutant protein or a KRAS G12D mutant protein) in a subject (e.g., a human subject), the method comprising administering to the subject (a) a CDR-H1 comprising (1) a glutamic acid residue (E) at its N-terminus, (2) a CDR-H1 comprising RAWMN (SEQ ID NO: 5), (3) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO: 6), (4) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 7), and (5) a V H (b) a CDR-L1 comprising (1) KSSQSVLYAGNNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10), Ldomain, and an effective amount of a small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor if the cancer is a KRAS G12C mutant cancer, or a KRAS G12D inhibitor if the cancer is a KRAS G12C mutant cancer). In some embodiments, a method is provided for stimulating phagocytosis of a population of cancer cells comprising one or more cancer cells expressing a KRAS mutant protein (e.g., a KRAS G12C mutant protein or a KRAS G12D mutant protein), the method comprising: (a) administering to the population an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 inhibitor comprising a CD47 domain and a CD47 residue at its C-terminus. H (b) a CDR-L1 comprising (1) KSSQSVLYAGNNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10), Ldomain, and an effective amount of a small molecule inhibitor (e.g., a KRAS G12C inhibitor, if the mutant protein is a KRAS G12C mutant protein, or a KRAS G12D inhibitor, if the mutant protein is a KRAS G12C mutant protein). In some embodiments, the KRAS mutant cancer (e.g., KRAS G12C mutant cancer or KRAS G12D mutant cancer), or the one or more cancer cells expressing a KRAS mutant protein (e.g., KRAS G12C mutant protein or KRAS G12D mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS mutant cancer is a KRAS G12C mutant cancer. In some embodiments, the population of cancer cells comprises one or more cells expressing a KRAS G12C mutant protein. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing KRAS G12C mutant protein) is NSCLC. In some embodiments, the KRAS mutant cancer is a KRAS G12D mutant cancer. In some embodiments, the population of cancer cells comprises one or more cells expressing KRAS G12D mutant protein. In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing KRAS G12D mutant protein) is a colorectal cancer, e.g., colorectal adenocarcinoma, or a colon cancer, e.g., colon adenocarcinoma. In some embodiments of the method of treatment, the anti-CD47 antibody (or an immunologically active fragment thereof) and a small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered simultaneously. In some embodiments of the treatment method, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS inhibitor (eg, a KRAS G12C inhibitor or a KRAS G12D inhibitor) are administered sequentially.In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor). In some embodiments of the method of treatment, the small molecule KRAS inhibitor (e.g., a KRAS G12C inhibitor or a KRAS G12D inhibitor) is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above. In some embodiments, the KRAS inhibitor is a KRAS G12D inhibitor. In some embodiments, the KRAS G12D inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12D inhibitor is MTRX1133, the structure of which is shown above.

[0108] In some embodiments, a method is provided for treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS G12C mutant protein) in a subject (e.g., a human subject), the method comprising administering to the subject (a) a CDR-H1 comprising (1) a glutamic acid residue (E) at its N-terminus, (2) a CDR-H1 comprising RAWMN (SEQ ID NO:5), (3) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO:6), (4) a CDR-H3 comprising SNRAFDI (SEQ ID NO:7), and (5) a CDR-H4 comprising a serine (S) at its C-terminus, H (b) a CDR-L1 comprising (1) KSSQSVLYAGNNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10), LThe method includes administering an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a CD47 domain and an effective amount of a small molecule KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer is NSCLC. In some embodiments of the treatment method, the anti-CD47 antibody (or an immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12C inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12C inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above.

[0109] In some embodiments, a method of treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS G12C mutant protein) in a subject (e.g., a human subject) is provided, the method comprising administering to the subject (a) a V H domain and (b) V comprising SEQ ID NO:2 L In some embodiments, a method is provided for stimulating phagocytosis of a population of cancer cells comprising one or more cancer cells expressing a KRAS G12C mutant protein, the method comprising administering to the population an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a KRAS G12C mutant domain, the KRAS G12C mutant domain, the V ... H domain and (b) V comprising SEQ ID NO:2L The method includes contacting an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a domain, and an effective amount of a small molecule KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing KRAS G12C mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing KRAS G12C mutant protein) is NSCLC. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12C inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12C inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above.

[0110] In some embodiments, a method is provided for treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS G12C mutant protein) in a subject (e.g., a human subject), the method comprising administering to the subject (a) a V H domain and (b) V comprising SEQ ID NO:2 LThe method includes administering an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a CD47 domain, and an effective amount of a small molecule KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer is NSCLC. In some embodiments of the method of treatment, the anti-CD47 antibody (or an immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12C inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12C inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above.

[0111] In some embodiments, a method is provided for treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS G12D mutant protein) in a subject (e.g., a human subject), the method comprising administering to the subject (a) a CDR-H1 comprising (1) a glutamic acid residue (E) at its N-terminus, (2) a CDR-H1 comprising RAWMN (SEQ ID NO:5), (3) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO:6), (4) a CDR-H3 comprising SNRAFDI (SEQ ID NO:7), and (5) a V H(b) a CDR-L1 comprising (1) KSSQSVLYAGNNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10), L The method includes administering an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a CD47 domain and an effective amount of a small molecule KRAS G12D inhibitor. In some embodiments, the KRAS G12D mutant cancer is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12D mutant cancer is colon cancer, e.g., colorectal adenocarcinoma, or colon cancer, e.g., colon adenocarcinoma. In some embodiments of the method of treatment, the anti-CD47 antibody (or an immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12D inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12D inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS G12D inhibitor is MRTX1122, the structure of which is shown above.

[0112] In some embodiments, a method is provided for treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS G12D mutant protein) in a subject (e.g., a human subject), the method comprising administering to the subject (a) a V H domain and (b) V comprising SEQ ID NO:2 LIn some embodiments, a method is provided for stimulating phagocytosis of a population of cancer cells comprising one or more cancer cells expressing a KRAS G12D mutant protein, the method comprising administering to the population an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a V domain comprising SEQ ID NO:1, and an effective amount of a small molecule KRAS G12D inhibitor. H domain and (b) V comprising SEQ ID NO:2 L The method includes contacting an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a domain, and an effective amount of a small molecule KRAS G12D inhibitor. In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing KRAS G12D mutant proteins) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing KRAS G12D mutant proteins) is colon cancer, e.g., colorectal adenocarcinoma, or colon cancer, e.g., colon adenocarcinoma. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12D inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12D inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS G12D inhibitor is MRTX1133, the structure of which is shown above.

[0113] In some embodiments, a method is provided for treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells that express a KRAS G12D mutant protein) in a subject (e.g., a human subject), the method comprising administering to the subject (a) a V H domain and (b) V comprising SEQ ID NO:2 L The method includes administering an effective amount of an anti-CD47 antibody (or an immunologically active fragment thereof) comprising a CD47 domain, and an effective amount of a small molecule KRAS G12D inhibitor. In some embodiments, the KRAS G12D mutant cancer is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12D mutant cancer is colon cancer, e.g., colorectal adenocarcinoma, or colon cancer, e.g., colon adenocarcinoma. In some embodiments of the method of treatment, the anti-CD47 antibody (or an immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12D inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12D inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof). In some embodiments, the KRAS G12D inhibitor is MRTX1122, the structure of which is shown above.

[0114] In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject), wherein the medicament is for administration together with a KRAS G12C inhibitor. In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject), wherein the medicament is for administration together with a KRAS G12C inhibitor. In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for stimulating phagocytosis of a population of cancer cells in a subject (e.g., a human subject), comprising one or more cancer cells expressing a KRAS G12C mutant protein, wherein the medicament is for administration together with a KRAS G12C inhibitor. In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for stimulating phagocytosis of a population of cancer cells in a subject (e.g., a human subject), comprising one or more cancer cells expressing a KRAS G12C mutant protein, wherein the medicament is for administration together with a KRAS G12C inhibitor. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing a KRAS G12C mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal cancer.In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing KRAS G12C mutant protein) is NSCLC. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is a polypeptide. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody of an antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) is any anti-CD47 antibody (or an immunologically active fragment thereof) described herein. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12C inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12C inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof).

[0115] In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12D mutant protein) in a subject (e.g., a human subject), wherein the medicament is for administration together with a KRAS G12D inhibitor. In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12D mutant protein) in a subject (e.g., a human subject), wherein the medicament is for administration together with a KRAS G12D inhibitor. In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for stimulating phagocytosis of a population of cancer cells in a subject (e.g., a human subject), comprising one or more cancer cells expressing a KRAS G12D mutant protein, wherein the medicament is for administration together with a KRAS G12D inhibitor. In some embodiments, there is provided a use of an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) in the manufacture of a medicament for stimulating phagocytosis of a population of cancer cells in a subject (e.g., a human subject), comprising one or more cancer cells expressing a KRAS G12D mutant protein, wherein the medicament is for administration together with a KRAS G12D inhibitor. In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing a KRAS G12D mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal cancer.In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing KRAS G12D mutant protein) is colorectal cancer, e.g., colorectal adenocarcinoma, or colon cancer, e.g., colon adenocarcinoma. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is a polypeptide. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody of an antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) is any anti-CD47 antibody (or an immunologically active fragment thereof) described herein. In some embodiments, the KRAS G12D inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12D inhibitor is MRTX1133, the structure of which is shown above. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered before the small molecule KRAS G12D inhibitor. In some embodiments of the method of treatment, the small molecule KRAS G12D inhibitor is administered before the anti-CD47 antibody (or immunologically active fragment thereof).

[0116] In some embodiments, there is provided a method for treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject), comprising an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the agent is for use (e.g., administered) together with a KRAS G12C inhibitor. In some embodiments, there is provided a method for treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject), comprising an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the KRAS G12C inhibitor is for use (e.g., administered) together with an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα). In some embodiments, there is provided a method for stimulating phagocytosis of a population of cancer cells, including one or more cancer cells expressing a KRAS G12C mutant protein, in a subject (e.g., a human subject), using an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the agent is for use (e.g., administered) with a KRAS G12C inhibitor. In some embodiments, there is provided a method for stimulating phagocytosis of a population of cancer cells, including one or more cancer cells expressing a KRAS G12C mutant protein, in a subject (e.g., a human subject), using an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the KRAS G12C inhibitor is for use (e.g., administered) with a KRAS G12C inhibitor.In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing the KRAS G12C mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing the KRAS G12C mutant protein) is NSCLC. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody of an antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) is any anti-CD47 antibody (or an immunologically active fragment thereof) described herein. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered prior to the small molecule KRAS G12C inhibitor.In some embodiments of the treatment method, a small molecule KRAS G12C inhibitor is administered prior to the anti-CD47 antibody (or an immunologically active fragment thereof).

[0117] In some embodiments, there is provided a method for treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12D mutant protein) in a subject (e.g., a human subject), comprising an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the agent is for use (e.g., administered) together with a KRAS G12D inhibitor. In some embodiments, there is provided a method for treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12D mutant protein) in a subject (e.g., a human subject), comprising an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the KRAS G12D inhibitor is for use (e.g., administered) together with an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα). In some embodiments, there is provided a method for stimulating phagocytosis of a population of cancer cells, comprising one or more cancer cells expressing KRAS G12D mutant protein, in a subject (e.g., a human subject), using an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the agent is for use (e.g., administered) together with a KRAS G12D inhibitor. In some embodiments, there is provided a method for stimulating phagocytosis of a population of cancer cells, comprising one or more cancer cells expressing KRAS G12D mutant protein, in a subject (e.g., a human subject), using an agent (e.g., a therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα), wherein the KRAS G12D inhibitor is for use (e.g., administered) together with a KRAS G12D inhibitor.In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing the KRAS G12D mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric-esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing the KRAS G12D mutant protein) is colon cancer, e.g., colorectal adenocarcinoma, or colon cancer, e.g., colon adenocarcinoma. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody of the antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) is any anti-CD47 antibody (or an immunologically active fragment thereof) described herein. In some embodiments, the KRAS G12D inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12D inhibitor is MRTX1133, the structure of which is shown above. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered prior to the small molecule KRAS G12D inhibitor.In some embodiments of the treatment method, a small molecule KRAS G12D inhibitor is administered prior to the anti-CD47 antibody (or an immunologically active fragment thereof).

[0118] In some embodiments, a composition (e.g., pharmaceutical composition) is provided that includes an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) for use in treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject), where the composition is for use (e.g., administered) together with a KRAS G12C inhibitor. In some embodiments, a composition (e.g., pharmaceutical composition) is provided that includes a KRAS G12C inhibitor for use in treating a KRAS G12C mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12C mutant protein) in a subject (e.g., a human subject), where the composition inhibitor is for use (e.g., administered) together with an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα). In some embodiments, a composition (e.g., pharmaceutical composition) is provided that includes an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) for use in stimulating phagocytosis of a population of cancer cells in a subject (e.g., human subject), comprising one or more cancer cells expressing a KRAS G12C mutant protein, wherein the composition is for use (e.g., administered) with a KRAS G12C inhibitor. In some embodiments, a composition is provided that includes KRAS G12C for stimulating phagocytosis of a population of cancer cells in a subject (e.g., human subject), comprising one or more cancer cells expressing a KRAS G12C mutant protein, wherein the composition is for use (e.g., administered) with an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα).In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing the KRAS G12C mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12C mutant cancer (or one or more cancer cells expressing the KRAS G12C mutant protein) is NSCLC. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody of an antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188 (also known as Retaplimab), IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) is any anti-CD47 antibody (or an immunologically active fragment thereof) described herein. In some embodiments, the KRAS G12C inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12C inhibitor is AMG 510, the structure of which is shown above. In some embodiments, the KRAS G12C inhibitor is MRTX849, the structure of which is shown above. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12C inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered prior to the small molecule KRAS G12C inhibitor.In some embodiments of the treatment method, a small molecule KRAS G12C inhibitor is administered prior to the anti-CD47 antibody (or an immunologically active fragment thereof).

[0119] In some embodiments, a composition (e.g., pharmaceutical composition) is provided that includes an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) for use in treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12D mutant protein) in a subject (e.g., a human subject), where the composition is for use (e.g., administered) together with a KRAS G12D inhibitor. In some embodiments, a composition (e.g., pharmaceutical composition) is provided that includes a KRAS G12D inhibitor for use in treating a KRAS G12D mutant cancer (i.e., a cancer comprising one or more cancer cells expressing a KRAS G12D mutant protein) in a subject (e.g., a human subject), where the composition inhibitor is for use (e.g., administered) together with an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα). In some embodiments, a composition (e.g., pharmaceutical composition) is provided that includes an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα) for use in stimulating phagocytosis of a population of cancer cells in a subject (e.g., human subject), comprising one or more cancer cells expressing a KRAS G12D mutant protein, wherein the composition is for use (e.g., administered) with a KRAS G12D inhibitor. In some embodiments, a composition is provided that includes KRAS G12D for stimulating phagocytosis of a population of cancer cells in a subject (e.g., human subject), comprising one or more cancer cells expressing a KRAS G12D mutant protein, wherein the composition is for use (e.g., administered) with an agent (e.g., therapeutic agent) that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα).In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing the KRAS G12D mutant protein) is lung cancer (e.g., lung adenocarcinoma or NSCLC), colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric-esophageal cancer, or rectal cancer. In some embodiments, the KRAS G12D mutant cancer (or one or more cancer cells expressing the KRAS G12D mutant protein) is colon cancer, e.g., colorectal adenocarcinoma, or colon cancer, e.g., colon adenocarcinoma. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody of the antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody (or an immunologically active fragment thereof). In some embodiments, the anti-CD47 antibody is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188 (also known as Retaplimab), IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing. In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) is any anti-CD47 antibody (or an immunologically active fragment thereof) described herein. In some embodiments, the KRAS G12D inhibitor is a small molecule inhibitor. In some embodiments, the KRAS G12D inhibitor is MRTX1133, the structure of which is shown above. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered simultaneously. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) and the small molecule KRAS G12D inhibitor are administered sequentially. In some embodiments of the method of treatment, the anti-CD47 antibody (or immunologically active fragment thereof) is administered prior to the small molecule KRAS G12D inhibitor.In some embodiments of the treatment method, a small molecule KRAS G12D inhibitor is administered prior to the anti-CD47 antibody (or an immunologically active fragment thereof).

[0120] Manufactured Products and Kits An article of manufacture is provided that includes materials useful for treating KRAS mutant cancer (e.g., KRAS G12C mutant cancer or KRAS G12D mutant cancer). In some embodiments, the KRAS mutant cancer (e.g., KRAS G12C mutant cancer or KRAS G12D mutant cancer) is lung adenocarcinoma, non-small cell lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric-esophageal cancer, or rectal adenocarcinoma). In certain embodiments, the article of manufacture or kit includes a container that contains a therapeutic agent that blocks the interaction between CD47 (e.g., hCD47) and SIRPα (e.g., hSIRPα). In some embodiments, such a therapeutic agent is a polypeptide, e.g., a polypeptide that binds to CD47 (e.g., hCD47). In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an antibody, an antibody construct, or an immunologically active fragment of an antibody or antibody construct. In some embodiments, the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody or an immunologically active fragment thereof. In some embodiments, the anti-CD47 antibody (or an immunologically active fragment thereof) is CC-90002 (also known as INBRX103), 5F9 (also known as Hu5F9-G4 and Magrolimab), LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, or ZL-1201. In some embodiments, the anti-CD47 antibody or an immunologically active fragment thereof is an anti-CD47 antibody (or a fragment thereof) described herein, or a pharmaceutical composition comprising such an anti-CD47 antibody or antibody fragment. In certain embodiments, the article of manufacture or kit comprises a container housing a nucleic acid(s) encoding an anti-CD47 antibody (or an immunologically active fragment thereof), e.g., an anti-CD47 antibody (or fragment thereof) described herein. In some embodiments, the kit comprises cells of a cell line that produces an anti-CD47 antibody (or an immunologically active fragment thereof) described herein. In some embodiments, the kit comprises one or more positive controls, e.g., CD47 (or a fragment thereof) or CD47 +In some embodiments, the kit includes a negative control, e.g., a surface or solution that is substantially free of CD47, or cells that do not express CD47.

[0121] In certain embodiments, the article of manufacture or kit comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV infusion bags, test tubes, and the like. The container can be formed from a variety of materials, such as glass or plastic. The container holds a composition (e.g., a composition comprising an anti-CD47 antibody or an immunologically active fragment thereof) that is effective, by itself or in combination with another composition, to treat (e.g., delay progression of) a KRAS mutant cancer (e.g., a KRAS G12C mutant cancer or a KRAS G12D mutant cancer), such as lung adenocarcinoma, non-small cell lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric-esophageal cancer, or rectal adenocarcinoma). The container may have a sterile access port (e.g., the container may be an intravenous fluid bag or a vial having a stopper pierceable by a hypodermic needle). At least one agent in the composition is an agent that blocks binding of hCD47 to hSIRPα. In some embodiments, at least one agent in the composition is an anti-CD47 antibody (or an immunologically active fragment thereof), e.g., an anti-CD47 antibody described herein. In some embodiments, the label or package insert indicates that the composition is used in combination with a KRAS G12C inhibitor to treat a KRAS G12C mutant cancer, such as lung adenocarcinoma, non-small cell lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric or esophageal cancer, or rectal adenocarcinoma. In some embodiments, the KRAS G12C mutant cancer is lung cancer, e.g., lung adenocarcinoma, or NSCLC. In some embodiments, the label or package insert indicates that the composition is used in combination with a KRAS G12D inhibitor to treat a KRAS G12D mutant cancer, such as lung adenocarcinoma, non-small cell lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric and esophageal cancer, or rectal adenocarcinoma.In some embodiments, the KRAS G12D mutant cancer is colorectal cancer, e.g., colorectal adenocarcinoma, or colon cancer, e.g., colon adenocarcinoma.

[0122] In some embodiments, the article of manufacture or kit is for the treatment of KRAS G12C cancer (e.g., a KRAS G12C cancer described herein) and includes (a) a first container containing a composition, the composition comprising an agent that blocks the binding of hCD47 to hSIRPα (such as an anti-CD47 antibody, or an immunologically active fragment thereof, e.g., an anti-CD47 antibody or fragment thereof described herein), and (b) a second container containing a composition, the composition comprising a KRAS G12C inhibitor (e.g., a polypeptide, antibody, fusion polypeptide, antisense oligonucleotide, or small molecule drug that can inhibit the activity of a KRAS G12C mutant protein). In some embodiments, the second container contains a small molecule KRAS G12C inhibitor. Exemplary small molecule KRAS G12C inhibitors that may be packaged by the kits provided herein include, but are not limited to, e.g., AMG 510 (Amgen), MRTX849 (Mirati), JAB-21822 (Jacobiopharma), GDC-6036 (Genentech), JDQ443 (Novartis), D-1553 (InventisBio and Merck Sharp & Dohme), GH35 (Genhouse Bio), GFH925 (GenFleet Therapeutics), BPI-421286 (Bettapharma), and LY3537982. In some embodiments, the article of manufacture or kit is for the treatment of KRAS G12D cancer (e.g., a KRAS G12D cancer described herein) and includes (a) a first container containing a composition, the composition comprising an agent that blocks the binding of hCD47 to hSIRPα (such as an anti-CD47 antibody, or an immunologically active fragment thereof, e.g., an anti-CD47 antibody or fragment thereof described herein), and (b) a second container containing a composition, the composition comprising a KRAS G12D inhibitor (e.g., a polypeptide, antibody, fusion polypeptide, antisense oligonucleotide, or small molecule drug that can inhibit the activity of a KRAS G12D mutant protein). In some embodiments, the second container contains a small molecule KRAS G12D inhibitor.Exemplary small molecule KRAS G12D inhibitors that may be packaged with the kits provided herein include, but are not limited to, MRTX1133 (Mirati). Additionally, the article of manufacture may further comprise additional containers containing pharma- ceutically acceptable buffers, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[0123] All publications and patent applications cited in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. EXAMPLES

[0124] The following examples are presented so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments described below are all or the only ones performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be allowed for. Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

[0125] Example 1. In vivo antitumor efficacy of anti-CD47 antibodies in combination with KRAS G12C inhibitors in mice bearing human non-small cell lung cancer (NSCLC) tumor xenografts The anti-tumor effect of anti-CD47 antibody TJC4 in combination with the KRAS G12C inhibitor AMG-510 was evaluated in mice bearing human NCI-H358 non-small cell lung cancer (NSCLC) tumor xenografts. The NCI-H358 cell line, established from a chemotherapy-naive NSCLC tumor, harbors a KRAS G12C mutation. TJC4 is a V-terminal fragment containing SEQ ID NO:1. H, V including SEQ ID NO:2 L and a human IgG4 Fc region containing a S228P substitution (EU numbering). AMG-510 is also known as sotorasib. KRAS G12C mutations, commonly found in NSCLC, lead to constitutive activation of the tumor growth-promoting RAS / MAPK signaling pathway.

[0126] Briefly, NCI-H358 cells (ATCC, CRL-5807) were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C. Thirty-two CB17 SCID mice (female, age: 6-8 weeks, body weight: 16-20 g) were then each cultured with 5 × 10 6 NCI-H358 cells (resuspended in 0.2 mL of DPBS containing 50% Matrigel) were inoculated subcutaneously on the right dorsal side. Seven days later, the tumor volume was 70–80 mm 3 Upon reaching this threshold, animals were randomly divided into four groups (n=8 / group) and administered (a) PBS (control), (b) AMG-510 monotherapy, (c) TJC4 monotherapy, or (d) AMG-510 in combination with TJC4 according to the dosages and frequency of administration shown in Table 1 below. [Table 1]

[0127] The tumor size of each animal was measured three times a week for 15 days. The tumor size was calculated as V = 0.5L × W 2 (where L and W represent the long and short diameters of the tumor, respectively). The average tumor size of the animals in each group was analyzed, as shown in Figure 1. The antitumor effect of the drug was also analyzed by TGI% (percentage of tumor growth inhibition). TGI% was calculated as follows: TGI(%)=[1-(V N -V0) / (V CN -V C0 )] × 100% (In the formula, V N is the mean tumor volume on day N of administration, V0 is the mean tumor volume on day 0 of administration, and V CNis the mean tumor volume in the control group on day N of administration, and V C0 is the mean tumor volume in the control group on day 0 of treatment).

[0128] T / C (relative tumor inhibition) was calculated as follows: T / C%=TRTV / CRTV×100% (where TRTV is the RTV of the treatment group and CRTV is the RTV of the control group). RTV=V N / V1 (where V1 is the mean tumor volume measured on day 1 of administration, and V N is the mean tumor volume measured on day N of administration).

[0129] As shown in Figure 1, mice treated with either the anti-CD47 antibody TJC4 as a single agent or the KRAS G12C inhibitor AMG-510 as a single agent did not show tumor growth inhibition compared to mice treated with a PBS control. In contrast, significant tumor growth inhibition was observed in mice treated with TJC4 in combination with AMG-510.

[0130] The synergy of the combination of TJC4 and AMG-510 was calculated as follows: Q=(TGI AMG-510+TJC4 / (T.G.I. AMG-510 +T.G.I. TJC4 -T.G.I. AMG-510 ×T.G.I. TJC4 ) (wherein Q≧1.15 indicates a synergistic effect between TJC4 and AMG-510, 0.85≦Q<1 indicates an additive effect between TJC4 and AMG-510, and Q<0.85 indicates an antagonistic effect between TJC4 and AMG-510.) On day 15, the Q value of TJC4 and AMG-510 was 3.33, indicating that the antitumor effect of the drug combination was synergistic.

[0131] On day 15, the mice were sacrificed and their tumor tissues were isolated, weighed, and dispersed into single-cell suspensions. The cells in each suspension were subjected to immunophenotyping by flow cytometry. The tumor weights of mice treated with TJC4 in combination with AMG-510 were found to be significantly lower than those of mice treated with either TJC4 as a single agent or AMG-510 as a single agent. See Figure 2. As shown in Figure 3A, tumors of mice treated with TJC4 in combination with AMG-510 showed significantly increased levels of calreticulin "CALR" (i.e., phagocytic signal) compared to tumors of mice treated with either drug as a single agent. As shown in Figures 3B and 3C, the M1:M2 ratio and M1 macrophage percentage were significantly increased in tumor cells of mice treated with TJC4 in combination with AMG-51 compared to tumor cells in mice given AMG 510 monotherapy. At the end of the study, tumor tissues were obtained from the mice, embedded in paraffin, and fixed in formalin. FFPE (formalin-fixed paraffin-embedded) samples were then subjected to tumor immune profiling via multiplex immunohistochemistry. As shown in Figures 3D and 3E, tumors from mice treated with TJC4 and AMG 510 showed significantly increased M1:M2 ratios and significantly decreased M2 macrophage percentages compared to tumors from mice treated with either drug as a single agent. Images of M1 and M2 macrophage populations in tumors from each treatment group are shown in Figure 3F (M2 macrophages are indicated by white arrows). Both M1 and M2 macrophages were F4 / 80 + and only M2 macrophages express CD206 + It is.

[0132] Example 2. Effect of a combination of anti-CD47 antibody and KRAS G12C inhibitor on phagocytosis of human non-small cell lung cancer (NSCLC) cells in vitro The following experiment was performed to evaluate the effect of anti-CD47 antibody TJC4 in combination with KRAS G12C inhibitor AMG-510 on in vitro phagocytosis of NCI-H358 human NSCLC tumor cells. As discussed in Example 1, NCI-H358 cells harbor a KRAS G12C mutation. In vitro phagocytosis assays were performed by co-culturing M0 macrophage cells with INCUCYTE® PHRODO®-labeled NCI-H358 tumor cells. Briefly, prior to co-culture, NCI-H358 cells were pretreated with AMG 510 at concentrations of 0 μm, 0.1 μm, 1 μm, or 10 μm for 24 hours. NCI-H358 cells were further incubated with M0 macrophages (at a 1:1 ratio) and 1 μg / ml TJC4 for 3 to 6 hours. NCI-H358 cells were then analyzed by an INCUCYTE® ZOOM live cell imaging system, and the percentage of phagocytosed NCI-H358 was calculated. Figures 4A-4C show that AMG 510 enhanced TJC4-mediated phagocytosis of NCI-H358 by macrophages.

[0133] The synergistic effect of the combination of AMG 510 and TJC4 in inducing phagocytosis was calculated as follows: F=(Combo / Blank) / ((C4 / Blank) x (SoC / Blank)).

[0134] An F value of ≧1.00 indicates synergy between AMG-510 and TJC4. As shown in Table 2, the combination of the anti-CD47 antibody TJC4 and the KRAS G12C inhibitor AMG-510 is synergistic in inducing phagocytosis of tumor cells at all concentrations of AMG-510 tested. [Table 2]

[0135] Example 3. Effect of a combination of anti-CD47 antibody and KRAS G12D inhibitor on phagocytosis of human colon cancer cells by macrophages in vitro LS180 human colon adenocarcinoma cells harboring a KRAS G12D mutation were incubated with (a) DMSO, (b) the KRAS G12D inhibitor MRTX1133 (100 nM), (c) the anti-CD47 antibody TJC4 (0.5 μg / ml), or (d) MRTX1133 (100 nM) + TJC4 (0.5 μg / ml) for 24 h and then labeled with 2.5 μM carboxyfluorescein succinimidyl ester (CFSE) for 10 min at 37° C. Macrophage cells obtained by coexisting human THP-1 monocytic leukemia cells with lipopolysaccharide (LPS) for 24 h were cultured in serum-free medium for 2 h and then 2 × 10 5 10,000 cells were added to CFSE-labeled LS180 cells. Macrophages and LS180 cells were co-cultured at 37°C for 2 hours. Macrophages were then stained with APC-labeled anti-F4 / 80 antibody, and CFSE+F4 / 80+ cells were detected by flow cytometry (10,000 cells were analyzed in each sample). The percentage of LS180 cells phagocytosed by macrophages was calculated by dividing the number of CFSE+F4 / 80+ cells (see the upper right quadrant of each flow cytometry histogram in Figure 5A, i.e., Q2) by the number of CFSE+ cells (see the upper left and upper right quadrants of each flow cytometry histogram in Figure 5A, i.e., Q1 and Q2, respectively). In other words, LS180 cells phagocytosed by macrophages (%) = [Q2 / (Q1+Q2)] x 100%. As shown in Figure 5B, both MRTX1133 as a single agent and TJC4 as a single agent increased the phagocytosis of LS180 cells by macrophages. The combination of MRTX1133 and TJC4 significantly increased the phagocytosis of LS180 cells by macrophages compared to either agent alone (see Figure 5B). The F value was 1.1, suggesting that MRTX1133 and TJC4 antibodies exhibited a synergistic effect on phagocytosis of G12D mutant colon tumor cells.

[0136] Example 4. Evaluation of the effect of anti-CD47 antibodies in combination with KRAS G12D inhibitors on apoptosis in human colon cancer cells LS180 human colon adenocarcinoma cells were incubated with (a) DMSO, (b) KRAS G12D inhibitor MRTX1133 (100 nM), (c) anti-CD47 antibody TJC4 (0.5 μg / ml), or (d) MRTX1133 (100 nM) + TJC4 (0.5 μg / ml) for 24 h and then labeled with fluorescein isothiocyanate (FITC)-annexin V and propidium iodide (PI). Annexin V (or annexin A5) is a member of the annexin family of intracellular proteins that binds phosphatidylserine (PS) in a calcium-dependent manner. PS is normally found only in the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and PS is translocated to the outer leaflet. Therefore, FITC-labeled annexin V can be used to specifically target and identify apoptotic cells. PI, a cell-impermeable nucleic acid intercalating dye. Following staining, cells were analyzed by flow cytometry. As shown in Figure 6A, both MRTX1133 as a single agent and TJC4 as a single agent increased the apoptosis of LS180 cells. The combination of MRTX1133 and TJC4 significantly increased the apoptosis of LS180 cells compared to either agent alone (see Figure 6B). The F value was 1.03, suggesting that MRTX1133 and TJC4 antibodies exhibited a synergistic effect on inducing apoptosis in G12D mutant colon tumor cells.

[0137] Example 5. In vivo antitumor efficacy of anti-CD47 antibodies in combination with KRAS G12D inhibitors in mice bearing human colon cancer tumor xenografts The antitumor efficacy of the anti-CD47 antibody TJC4 in combination with the KRAS G12D inhibitor MRTZ1133 was evaluated in mice bearing human LS180 colon adenocarcinoma tumor xenografts. 5LS180 cells were injected into the cecal wall of BALB / c nude mice. After 2 weeks, tumors formed and grew stably. Mice were given one of the following treatments for 4 weeks: (a) PBS (control), (b) TJC4 (1 mg / kg, intraperitoneally twice a week), (c) MRTX1133 (20 mg / kg, intraperitoneally twice a week), or (d) MRTX1133 (20 mg / kg, intraperitoneally twice a week) + TJC4 (1 mg / kg, intraperitoneally twice a week). See Figure 7A. Tumor growth was assessed weekly by bioluminescence imaging (BLI). As shown in Figure 7B, MRTX1133 as a single agent inhibited tumor growth. TJC4 as a single agent had a weak inhibitory effect on tumor growth. The combination of MRTX1133 and TJC4 showed increased tumor growth inhibition compared to either agent alone. Notably, the combined treatment resulted in complete tumor regression (see Figure 7C). The synergy coefficient Q was 2.4492, suggesting that MRTX1133 and TJC4 antibodies act synergistically to suppress tumor growth.

[0138] The present invention has been described in terms of specific embodiments found or proposed by the inventors to include preferred modes for carrying out the invention. In light of this disclosure, those skilled in the art will appreciate that numerous modifications and changes can be made in the specific embodiments exemplified without departing from the intended scope of the invention. For example, due to codon redundancy, modifications can be made in the underlying DNA sequence without affecting the protein sequence. Furthermore, due to biological functional equivalence considerations, modifications can be made in the protein structure without affecting the type or amount of biological action. All such modifications are intended to be included within the scope of the appended claims. [Table 5-1] [Table 5-2] [Table 5-3]

Claims

1. A method of treating cancer in a subject, comprising administering to the subject an effective amount of an agent that blocks the interaction between CD47 and SIRPα and an effective amount of a KRAS inhibitor, wherein the cancer comprises one or more cancer cells that express a KRAS mutant protein.

2. 1. A method for stimulating phagocytosis of a population of cancer cells by macrophages, comprising contacting the population with an effective amount of an agent that blocks the interaction between CD47 and SIRPα, and an effective amount of a KRAS inhibitor, wherein the population of cancer cells comprises one or more cancer cells that express a KRAS mutant protein.

3. The method of claim 1 or 2, wherein the agent that blocks the interaction between CD47 and SIRPα is a polypeptide that binds to CD47.

4. The method of claim 3, wherein the polypeptide that binds to CD47 is an anti-CD47 antibody or an immunologically active fragment thereof.

5. 5. The method of claim 4, wherein the anti-CD47 antibody or the immunologically active fragment thereof is CC-90002, 5F9, LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment of any one of the foregoing.

6. The anti-CD47 antibody or the immunologically active fragment thereof comprises a heavy chain variable domain (V Ｈ ) and a light chain variable domain (V) as defined in SEQ ID NO:

2. Ｌ 5. The method of claim 4, comprising the three CDRs:

7. The anti-CD47 antibody or the immunologically active fragment thereof, (a) a V comprising (1) a CDR-H1 comprising RAWMN (SEQ ID NO: 5), (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO: 6), and (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 7), Ｈ and, (b) V comprising (1) a CDR-L1 comprising KSSQSVLYAGNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10). Ｌ and wherein the CDR sequences are defined according to the Kabat numbering system.

8. The V of the anti-CD47 antibody or the immunologically active fragment thereof Ｈ The V domain of the anti-CD47 antibody or the immunologically active fragment thereof comprises an amino acid sequence having at least 95% identity to SEQ ID NO:

1. Ｌ The method of claim 6 or 7, wherein said sequence comprises an amino acid sequence having at least 95% identity to SEQ ID NO:

2.

9. The V Ｈ The N-terminal amino acid of the domain is E, Ｈ The method according to any one of claims 6 to 8, wherein the C-terminal amino acid of the domain is S.

10. (a) the V according to the Kabat numbering system; Ｈ The N-terminal amino acid of the domain corresponds to position H1, Ｈ The C-terminal amino acid of the domain corresponds to position H113. (b) said V according to the Chothia numbering system; Ｈ The N-terminal amino acid of the domain corresponds to position H1, Ｈ the C-terminal amino acid of the domain corresponds to position H113, or (c) said V according to the IMGT numbering system; Ｈ The N-terminal amino acid of the domain corresponds to position H1, Ｈ The C-terminal amino acid of the domain corresponds to position H128.

10. The method of claim 9.

11. The V Ｈ The N-terminal amino acid of the domain corresponds to amino acid 1 of SEQ ID NO:1, Ｈ The method of any one of claims 6 to 10, wherein the C-terminal amino acid of the domain corresponds to amino acid 118 of SEQ ID NO:

1.

12. The method of any one of claims 4 to 11, wherein the anti-CD47 antibody is a full-length antibody.

13. 13. The method of claim 12, wherein the full length anti-CD47 antibody comprises a human IgG4 Fc region or a variant thereof comprising a S228P substitution, and the amino acid numbering is according to the EU index.

14. The method of claim 12 or 13, wherein the full length anti-CD47 antibody comprises a heavy chain comprising SEQ ID NO:3 or SEQ ID NO:35 and a light chain comprising SEQ ID NO:

4.

15. The method according to any one of claims 1 to 14, wherein the KRAS inhibitor is a KRAS G12C inhibitor and the KRAS mutant protein is a KRAS G12C mutant protein.

16. 16. The method of claim 15, wherein the KRAS G12C inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of the KRAS G12C mutant protein.

17. 17. The method of claim 16, wherein the KRAS G12C inhibitor is a small molecule.

18. 18. The method of claim 17, wherein the small molecule is selected from the group consisting of AMG 510, MRTX849, JAB-21822, GDC-6036, JDQ443, D-1553, GH35, GFH925, BPI-421286, and LY3537982.

19. 19. The method of claim 18, wherein the small molecule is AMG 510 or MRTX849.

20. The method according to any one of claims 1 to 14, wherein the KRAS inhibitor is a KRAS G12D inhibitor and the KRAS mutant protein is a KRAS G12D mutant protein.

21. 21. The method of claim 20, wherein the KRAS G12D inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of the KRAS G12D mutant protein.

22. 22. The method of claim 21, wherein the KRAS G12D inhibitor is a small molecule.

23. 23. The method of claim 22, wherein the small molecule is MTRX1122.

24. The method according to any one of claims 1 and 3 to 23, wherein the cancer is lung cancer, colon cancer, colorectal cancer, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric / esophageal cancer, or rectal cancer.

25. 25. The method of any one of claims 1, 2-19, and 24, wherein the cancer is KRAS G12C cancer.

26. 26. The method of claim 25, wherein the KRAS G12C cancer is lung cancer.

27. 27. The method of claim 26, wherein the lung cancer is lung adenocarcinoma or non-small cell lung cancer (NSCLC).

28. 28. The method of claim 27, wherein the lung cancer is non-small cell lung cancer (NSCLC).

29. The method of any one of claims 1 to 14 and 20 to 24, wherein the cancer is a KRAS G12D cancer.

30. 30. The method of claim 29, wherein the KRAS G12D cancer is colorectal or colon cancer.

31. The method of any one of claims 4 to 30, wherein the anti-CD47 antibody or the immunologically active fragment thereof and the KRAS inhibitor are administered simultaneously.

32. The method of any one of claims 4 to 31, wherein the anti-CD47 antibody or the immunologically active fragment thereof and the KRAS inhibitor are administered sequentially.

33. The method of claim 32, wherein the anti-CD47 antibody, or the immunologically active fragment thereof, is administered prior to the KRAS inhibitor.

34. The method of claim 32, wherein the KRAS inhibitor is administered prior to the anti-CD47 antibody or the immunologically active fragment thereof.

35. 35. The method of any one of claims 1 and 34, wherein the subject is a human.

36. The method of any one of claims 2 to 24, wherein the one or more cancer cells expressing a KRAS mutant protein are lung cancer cells, colon cancer cells, colorectal cancer cells, pancreatic cancer cells, cholangiocarcinoma cells, endometrial cancer cells, ovarian cancer cells, peritoneal cancer cells, bladder cancer cells, gastric cancer cells, thyroid cancer cells, melanoma cells, breast cancer cells, head and neck cancer cells, multiple myeloma cells, acute myeloid leukemia (AML) cells, uterine cancer cells, gastric / esophageal cancer cells, or rectal cancer cells.

37. 37. The method of claim 36, wherein the KRAS inhibitor is a KRAS G12C inhibitor, the one or more cancer cells expressing a KRAS mutant protein express a KRAS G12C mutant protein, the one or more cancer cells expressing the KRAS G12C mutant protein are lung cancer cells, and the lung cancer cells are lung adenocarcinoma cells or non-small cell lung cancer (NSCLC) cells.

38. 38. The method of claim 37, wherein the lung cancer cells are NSCLC cells.

39. 37. The method of claim 36, wherein the KRAS inhibitor is a KRAS G12D inhibitor, the one or more cancer cells expressing a KRAS mutant protein express a KRAS G12D mutant protein, and the one or more cancer cells expressing a KRAS G12D mutant protein are colon cancer cells or colorectal cancer cells.

40. 1. A kit for treating cancer in a human subject, comprising: (a) an agent that blocks the interaction between CD47 and SIRPα; (b) instructions for administering to a subject having a cancer, the cancer cell comprising one or more cancer cells that express a KRAS mutant protein, an effective amount of the agent and an effective amount of a KRAS inhibitor; and The kit comprising:

41. The kit of claim 40, wherein the polypeptide is a polypeptide that binds to CD47.

42. The kit of claim 41 , wherein the polypeptide that binds to CD47 is an anti-CD47 antibody or an immunologically active fragment thereof.

43. The anti-CD47 antibody or the immunologically active fragment thereof, (a) a V comprising (1) a CDR-H1 comprising RAWMN (SEQ ID NO: 5), (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO: 6), and (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 7), Ｈ and, (b) V comprising (1) a CDR-L1 comprising KSSQSVLYAGNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10). Ｌ and wherein the CDR sequences are defined according to the Kabat numbering system.

44. The V of the anti-CD47 antibody or the immunologically active fragment thereof Ｈ The V domain of the anti-CD47 antibody or the immunologically active fragment thereof comprises an amino acid sequence having at least 95% identity to SEQ ID NO:

1. Ｌ The kit of claim 43, wherein said kit comprises an amino acid sequence having at least 95% identity to SEQ ID NO:

2.

45. The V Ｈ The N-terminal amino acid of the domain is E, Ｈ 45. The kit of claim 43 or 44, wherein the C-terminal amino acid of the domain is S.

46. The kit of any one of claims 42 to 45, wherein the anti-CD47 antibody is a full-length antibody.

47. The kit of claim 46, wherein the full length anti-CD47 antibody comprises a heavy chain comprising SEQ ID NO:3 or SEQ ID NO:35 and a light chain comprising SEQ ID NO:

4.

48. The kit of any one of claims 40 to 47, further comprising said KRAS inhibitor.

49. 49. The kit of claim 48, wherein the KRAS inhibitor is a KRAS G12C inhibitor and the cancer comprises one or more cancer cells expressing a KRASG12C mutant protein.

50. 50. The kit of claim 49, wherein the KRAS G12C inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of the KRAS G12C mutant protein.

51. 51. The kit of claim 50, wherein the KRAS G12C inhibitor is a small molecule inhibitor.

52. 52. The kit of claim 51, wherein the small molecule is selected from the group consisting of AMG 510, MRTX849, JAB-21822, GDC-6036, JDQ443, D-1553, GH35, GFH925, BPI-421286, and LY3537982.

53. 53. The kit of claim 52, wherein the small molecule is AMG 510 or MRTX849.

54. 49. The kit of claim 48, wherein the KRAS inhibitor is a KRAS G12D inhibitor and the cancer comprises one or more cancer cells expressing a KRASG12D mutant protein.

55. 55. The kit of claim 54, wherein the KRAS G12D inhibitor is an antibody, peptide, protein, antisense oligonucleotide, or small molecule that inhibits the activity of the KRAS G12D mutant protein.

56. 56. The kit of claim 55, wherein the KRAS G12D inhibitor is a small molecule inhibitor.

57. 57. The kit of claim 56, wherein the small molecule is MRTX1133.

58. The kit of any one of claims 40 to 57, wherein the cancer comprising one or more cancer cells expressing a KRAS mutant protein is lung cancer, colon adenocarcinoma, colorectal adenocarcinoma, pancreatic cancer, bile duct cancer, endometrial cancer, ovarian cancer, peritoneal cancer, bladder cancer, gastric cancer, thyroid cancer, melanoma, breast cancer, head and neck cancer, multiple myeloma, acute myeloid leukemia (AML), uterine cancer, gastric / esophageal cancer, or rectal adenocarcinoma.

59. A pharmaceutical composition comprising an agent that blocks the interaction between CD47 and SIRPα and a KRAS inhibitor.

60. 60. The pharmaceutical composition of claim 59, wherein the agent that blocks the interaction between CD47 and SIRPα is an anti-CD47 antibody or an immunologically active fragment thereof.

61. 61. The pharmaceutical composition of claim 60, wherein the anti-CD47 antibody or immunologically active fragment thereof is CC-90002, 5F9, LQ001, HLX24, TI-061, AO-176, SRF-231, IBI-188, IMC-002, SHR-1603, STI-6643, ZL-1201, or an immunologically active fragment.

62. The anti-CD47 antibody or the immunologically active fragment thereof, (a) a V comprising (1) a CDR-H1 comprising RAWMN (SEQ ID NO: 5), (2) a CDR-H2 comprising RIKRKTDGETTDYAAPVKG (SEQ ID NO: 6), and (3) a CDR-H3 comprising SNRAFDI (SEQ ID NO: 7), Ｈ and, (b) V comprising (1) a CDR-L1 comprising KSSQSVLYAGNRNYLA (SEQ ID NO: 8), (2) a CDR-L2 comprising QASTRAS (SEQ ID NO: 9), and (3) a CDR-L3 comprising QQYYTPPLA (SEQ ID NO: 10). Ｌ and wherein the CDR sequences are defined according to the Kabat numbering system.

63. The pharmaceutical composition of any one of claims 58 to 62, wherein the KRAS inhibitor is a KRAS G12C inhibitor.

64. 64. The pharmaceutical composition of claim 63, wherein the KRAS G12C inhibitor is selected from the group consisting of AMG 510, MRTX849, JAB-21822, GDC-6036, JDQ443, D-1553, GH35, GFH925, BPI-421286, and LY3537982.

65. The pharmaceutical composition of any one of claims 58 to 62, wherein the KRAS inhibitor is a KRAS G12D inhibitor.

66. 66. The pharmaceutical composition of claim 65, wherein the KRAS G12D inhibitor is MTRX1133.

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