Combination therapy for cancer using anti-CD24 antibody and PD-1-PD-L1 pathway blocking antibody

Abstract

The present application relates to a composition comprising i) an antibody, or an antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or an antigen-binding portion thereof, capable of inhibiting the binding / interaction of PD-1 and PD-L1, and the use of the composition in the treatment of cancer. The present application also relates to a combination cancer therapy using i) an antibody, or antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or antigen-binding portion thereof, capable of inhibiting the binding / interaction of PD-1 and PD-L1.

Description

[Technical Field]

[0001] This application claims priority from Chinese Patent Application No. 202211568698.8, filed on December 7, 2022.

[0002] This application relates to the combination of an anti-CD24 antibody, or antigen-binding portion thereof, and an antibody, or antigen-binding portion thereof, capable of inhibiting the binding / interaction of PD-1 and PD-L1 in the treatment of cancer. Specifically, this application relates to a composition comprising i) an antibody, or antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or antigen-binding portion thereof, capable of inhibiting the binding / interaction of PD-1 and PD-L1, and the use of the composition in the treatment of cancer and / or the preparation of a medicament for the treatment of cancer. [Background technology]

[0003] Cancer cells have developed numerous mechanisms to evade host immune surveillance, including: 1) evading immune surveillance by macrophages, T lymphocytes, B lymphocytes, and natural killer cells by highly expressing the membrane protein CD24, which binds to receptors such as Siglec-10 on the surface of immune cells, thereby suppressing immune activation; and 2) evading immune surveillance and inducing apoptosis of immune cells by expressing high levels of the membrane proteins PD-L1 and PD-L2, which bind to PD-1 on the surface of immune cells such as T cells. As described above, cancer cells are highly intelligent and can rapidly proliferate based on the evasion mechanisms they have developed. CD24 and Siglec-10

[0004] Sialic acid-binding immunoglobulin-like lectins (Siglecs) are immunoglobulin-like type I transmembrane proteins. Among the Siglec family, Siglec-10 is an inhibitory receptor widely expressed on immune cells, such as macrophages, B cells, NK cells, and activated T cells. It has five extracellular Ig-like domains, a transmembrane region, and a cytoplasmic tail. The IgV domain of Siglec-10 contains a key arginine residue involved in sialic acid recognition. Expression of Siglec-10 on T cells is known to inhibit T cell activation by suppressing the formation of T cell major histocompatibility complex class I (MHC-I) peptide complexes and the phosphorylation of T cell receptor-associated kinases, Lck, and ZAP-70. Siglec-10, expressed on B cells and NK cells, can suppress BCR-mediated and NK cell receptor-mediated signaling (Yin et al. (2020) Front Immunol 11:1324).

[0005] CD24 is a glycosylphosphatidylinositol-anchored protein found on the surface of developing T lymphocytes and most B lymphocytes (Yin et al., supra). Numerous cancer cells also highly express CD24, including ovarian cancer, breast cancer, cervical cancer, endometrial cancer, colorectal cancer, non-small cell lung cancer, small cell lung cancer, head and neck tumors, urothelial carcinoma, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), multiple myeloma, Hodgkin's lymphoma (HL), non-Hodgkin's lymphoma (NHL), cholangiocarcinoma, liver cancer, bladder cancer, pancreatic cancer, gastric cancer, and glioblastoma (Barkal et al., (2019) Nature 572(7769):392-396; Liu et al., (2013) Oncol Lett 6(1):96-100; Panagiotou E et al., (2022) J Pers Med. 12(8):1235). CD24 on cancer cells interacts with Siglec-10 on immune cells, generating a "don't eat me" signal that is used for immune evasion and protects tumor cells from immune attack, including inhibiting phagocytosis by macrophages.

[0006] Studies have shown a correlation between CD24 expression and bladder tumor recurrence (Liu et al., supra). In patients with ovarian cancer, CD24 expression is an independent predictor of overall survival and correlates with tumor stage, peritoneal, and lymph node metastasis; CD24-positive cells are associated with increased proliferation of a highly invasive phenotype and cisplatin resistance in ovarian cancer cells (Nakamura et al., (2017) Oncol Rep. 37(6):3189-3200). Monoclonal anti-CD24 antibodies have been shown to reduce lung metastasis and prolong overall survival in mouse models of bladder cancer and triple-negative breast cancer. It has also been shown in the literature that antibody-mediated inhibition of CD24-Siglec10 interaction can induce a reduction in macrophage-mediated tumor growth and prolong the survival of tumor-bearing mice (Barkal et al., supra; Chan et al., (2019) Mol Cancer Ther 18(1):147-161; Overdevest et al., (2011) Cancer Res 71(11):3802-11). In addition, in various preclinical animal studies, CD24 inhibitors have shown inhibitory effects on a wide range of tumor models, including ovarian cancer, breast cancer, non-small cell lung cancer, small cell lung cancer, head and neck tumors, urothelial carcinoma, non-Hodgkin's lymphoma (NHL), liver cancer, bladder cancer, pancreatic cancer, gastric cancer, and colorectal cancer (Panagiotou E et al., (2022) J Pers Med. 12(8):1235). PD-L1 and PD-1

[0007] PD-1 is a checkpoint molecule that downregulates immune responses and is primarily expressed in activated T cells and B cells. PD-L1 and PD-L2 are two ligands for PD-1, with PD-L1 being expressed, among other things, on antigen-presenting cells, T cells, B cells, monocytes, and epithelial cells (Keir ME et al., (2008) Annu Rev Immunol. 26:677-704; Chen J et al., (2016) Ann Oncol. 27(3):409-416). Engagement of PD-L1 or PD-L2 with PD-1 transmits an inhibitory signal that prevents activation of T cells and B cells, achieving immune system homeostasis and avoiding unintended damage to normal cells.

[0008] However, PD-1 signaling can be exploited by tumor cells to help them evade immune surveillance. Studies have shown that various tumor cells constitutively express PD-L1 and / or PD-L2, and that the tumor microenvironment (TME) induces immune cells, such as infiltrating T cells, to highly express PD-1 molecules. In this situation, PD-1 signaling in the tumor microenvironment is continuously activated, suppressing T cell-mediated killing of tumor cells.

[0009] Studies suggest that blocking PD-1 signaling may suppress the growth of various solid and hematological tumors. Approved or clinically tested PD-1 or PD-L1 targeting indications include mantle cell lymphoma, diffuse large B-cell lymphoma, and follicular lymphoma, as well as melanoma, non-small cell lung cancer, renal cell carcinoma, Hodgkin's lymphoma, bladder cancer, head and neck cancer, neuroendocrine tumors, and solid tumors with high microsatellite instability and defective mismatch repair (Akinleye A, Rasool Z. (2019) J Hematol Oncol. 12(1):92; Chong Sun et al. (2018) Immunity 48(3):434-452). Anti-PD-1 antibodies currently on the market include tislelizumab, nivolumab (Opdivo), or pembrolizumab (Keytruda), and approved anti-PD-L1 antibodies include atezolizumab, durvalumab, and avelumab. Combination therapy

[0010] Although therapies targeting single molecules such as PD-1 or PD-L1 have shown efficacy in a wide range of cancers, a significant proportion of patients do not respond to them. For example, several papers have noted that the efficacy of PD-1 or PD-L1 blocking antibodies is limited in ovarian and pancreatic cancers (Panagiotou E et al., (2022) supra), while CD24 is highly expressed on ovarian and pancreatic cancers (Barkal et al., (2019) Nature 572(7769):392-396; Liu et al., (2013) Oncol Lett 6(1):96-100).

[0011] Drug combinations have become a new therapeutic strategy, where targeting more than one signaling pathway can more efficiently kill cancer cells and suppress cancer growth. However, not all drug combinations lead to better therapeutic outcomes, and not all drugs can be combined for use. For example, in a pooled analysis of 14 Phase I-III trials, a 93% incidence of side effects was observed in patients receiving dual therapy with anti-PD-1 antibodies and anti-CTLA-4 antibodies, while 64% of patients receiving multiple doses of anti-CTLA-4 antibodies alone experienced immune-related side effects (Wolchok, JD et al., (2013) N. Engl. J. Med. 369:122-33). As another example, the combination of panobinostat and carfilzomib caused a 2% increase in treatment-related heart failure and treatment-related death in patients with relapsed / refractory multiple myeloma (Berdeja JG et al. (2015) Haematologica 100(5):670-676).

[0012] In addition, the interactions and roles of the two immune checkpoint pathways, CD24 / Siglec-10 and PD-1 / PD-L1, have not yet been elucidated, and more data are needed to support the anticancer effects of combining drugs that target these two axes (Panagiotou E et al., (2022), see above). Interestingly, in non-small cell lung cancer (NSCLC) with relatively low PD-L1 expression, CD24 was clearly found to be inversely correlated with progression-free survival in patients treated with immune checkpoint inhibitors (Ozawa Y et al., (2021) Cancer Sci. 112(1):72-80).

[0013] Citation or identification of any document in this application is not an admission that such document is available as prior art to the present invention. Summary of the Invention

[0014] The inventors of the present application have surprisingly found that by simultaneously targeting the CD24 pathway and the PD-1-PD-L1 pathway, for example by using an antibody that specifically binds CD24 in combination with an antagonist anti-PD-1 or anti-PD-L1 antibody, tumor or cancer growth can be synergistically inhibited.

[0015] In a first aspect, the present application provides: i) an antibody, or an antigen-binding portion thereof, capable of specifically binding to CD24; and ii) A method of treating cancer comprising administering to a subject an antibody, or an antigen-binding portion thereof, capable of inhibiting the binding of PD-1 to PD-L1 or the interaction of PD-1 and PD-L1.

[0016] The antibody or antigen-binding portion thereof capable of specifically binding to CD24 specifically binds to CD24 (e.g., human CD24) and preferably inhibits CD24-Siglec binding or CD24-Siglec interaction. In some embodiments, the antibody or antigen-binding portion thereof capable of specifically binding to CD24 has an FcR-binding heavy chain constant region and inhibits CD24. + It can induce antibody-dependent cell-mediated cytotoxicity (ADCC) against tumor cells.

[0017] In some embodiments, the antibody, or antigen-binding portion thereof, capable of specifically binding to CD24 has a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises heavy chain variable region CDR1 (HV-CDR1), HV-CDR2, and HV-CDR3; and the light chain variable region comprises light chain variable region CDR1 (LV-CDR1), LV-CDR2, and LV-CDR3, wherein HV-CDR1, HV-CDR2, and HV-CDR3 are at least nucleotides selected from the group consisting of GYSITSGYS (SEQ ID NO: 1), IHYSGST (SEQ ID NO: 2), and ARGADYALDY (SEQ ID NO: 3), respectively. LV-CDR1, LV-CDR2, and LV-CDR3 may each comprise an amino acid sequence having 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to QSLLYSSNQKNY (SEQ ID NO: 4), WAS, and QQNFIYPLT (SEQ ID NO: 5), respectively. In certain embodiments, the heavy and light chain variable regions may comprise amino acid sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to 1) SEQ ID NOs: 6 and 7, respectively; 2) SEQ ID NOs: 6 and 10, respectively; or 3) SEQ ID NOs: 11 and 12, respectively. In one embodiment, the heavy and light chain variable regions may comprise amino acid sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 6 and 7, respectively. The antibody or antigen-binding portion thereof capable of specifically binding to CD24 may further comprise a heavy chain constant region and / or a light chain constant region. The heavy chain constant region comprises FcR binding ability and can induce, for example, ADCC. In one embodiment, the heavy chain constant region may be, for example, a human IgG1 heavy chain constant region comprising the amino acid sequence of SEQ ID NO:8.The light chain constant region can be, for example, a human K light chain constant region comprising, for example, the amino acid sequence of SEQ ID NO:9.

[0018] An antibody or antigen-binding portion thereof capable of inhibiting the binding of PD-1 to PD-L1 or the PD-1-PD-L1 interaction can be an antagonist anti-PD-1 antibody or antigen-binding portion thereof capable of specifically binding to PD-1, or an antagonist anti-PD-L1 antibody or antigen-binding portion thereof capable of specifically binding to PD-L1.

[0019] The antagonist anti-PD-1 antibody, or antigen-binding portion thereof, may specifically bind to PD-1 (e.g., human PD-1) and preferably inhibit the binding or interaction of PD-1 and PD-L1, or the PD-1-PD-L1 and PD-1-PD-L2 binding or interaction. In certain embodiments, the antagonist anti-PD-1 antibody, or antigen-binding portion thereof, may be tislelizumab, nivolumab (e.g., Opdivo®), pembrolizumab (e.g., Keytruda®), or an antigen-binding portion thereof.

[0020] The antagonist anti-PD-L1 antibody, or antigen-binding portion thereof, may specifically bind to PD-L1 (e.g., human PD-L1) and preferably inhibit the binding of PD-1 to PD-L1 or the PD-1-PD-L1 interaction. In one embodiment, the antagonist anti-PD-L1 antibody, or antigen-binding portion thereof, may bind an FcR, e.g., PD-L1 + It may have a heavy chain constant region capable of inducing ADCC against tumor cells. The antagonist anti-PD-L1 antibody, or antigen-binding portion thereof, may be atezolizumab (e.g., Tecentriq®), durvalumab, avelumab, IMM2515H, or an antigen-binding portion thereof.

[0021] IMM2515H has a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain variable region CDR1 (HV-CDR1), HV-CDR2, and HV-CDR3, and the light chain variable region comprises a light chain variable region CDR1 (LV-CDR1), LV-CDR2, and LV-CDR3, and the HV-CDR1, HV-CDR2, and HV-CDR3 are at least 85%, 86%, and 87% identical to GYTFTSNW (SEQ ID NO: 13), IHPNSGSS (SEQ ID NO: 14), and ARSYYGSSPYYFDY (SEQ ID NO: 15), respectively. , 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to QDIINY (SEQ ID NO: 16), YTS, and QQGDTLPWT (SEQ ID NO: 17), respectively, and LV-CDR1, LV-CDR2, and LV-CDR3 may comprise amino acid sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to QDIINY (SEQ ID NO: 16), YTS, and QQGDTLPWT (SEQ ID NO: 17), respectively. In certain embodiments, the heavy chain variable region and the light chain variable region may comprise amino acid sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 18 and 19, respectively. IMM2515H may further comprise a heavy chain constant region and / or a light chain constant region. The heavy chain constant region may have FcR binding ability and, for example, induce ADCC. In one embodiment, the heavy chain constant region may be, for example, a human IgG1 heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 8. The light chain constant region may be, for example, a human K light chain constant region comprising the amino acid sequence of SEQ ID NO: 9.

[0022] The method may comprise the steps of simultaneously administering i) an antibody, or antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or antigen-binding portion thereof, capable of inhibiting the binding of PD-1 to PD-L1 or the interaction of PD-1 and PD-L1, or the steps of sequentially administering i) an antibody, or antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or antigen-binding portion thereof, capable of inhibiting the binding of PD-1 to PD-L1 or the interaction of PD-1 and PD-L1.

[0023] In some embodiments, the cancer may be a solid tumor or a hematological cancer, including, but not limited to, gastric cancer, liver cancer, colon cancer, non-small cell lung cancer, small cell lung cancer, ovarian cancer, pancreatic cancer, breast cancer, cervical cancer, endometrial cancer, colorectal cancer, head and neck tumors, urothelial cancer, cholangiocarcinoma, bladder cancer, glioblastoma, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), multiple myeloma, Hodgkin's lymphoma (HL), and non-Hodgkin's lymphoma (NHL). In some embodiments, the cancer may be a CD24-expressing cancer. In some embodiments, the cancer may be a cancer that does not express CD24. In some embodiments, the cancer may be a cancer that expresses PD-L1 and / or PD-L2. In some embodiments, the cancer may be a cancer that does not express PD-L1 or PD-L2. In some embodiments, the cancer may be a cancer that highly expresses PD-L1. In some embodiments, the cancer may be a cancer that has low PD-L1 expression. In some embodiments, the cancer may be colon cancer.

[0024] The present application also relates to the use of i) antibodies, or antigen-binding portions thereof, capable of specifically binding to CD24, and ii) antibodies, or antigen-binding portions thereof, capable of inhibiting the binding of PD-1 to PD-L1 or the PD-1-PD-L1 interaction, in the treatment of cancer and in the preparation of a medicament for treating cancer.

[0025] In a second aspect, the present application provides a composition, such as a pharmaceutical composition, comprising i) an antibody, or an antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or an antigen-binding portion thereof, capable of inhibiting the binding of PD-1 to PD-L1 or the interaction of PD-1 and PD-L1.

[0026] The antibody, or antigen-binding portion thereof, capable of specifically binding to CD24, and the antibody, or antigen-binding portion thereof, capable of inhibiting the binding of PD-1 to PD-L1 or the interaction of PD-1 and PD-L1 are as defined above.

[0027] In one embodiment, a pharmaceutical composition is provided comprising therapeutically effective amounts of i) an antibody or antigen-binding portion thereof capable of specifically binding to CD24, and ii) an antibody or antigen-binding portion thereof capable of inhibiting the binding of PD-1 to PD-L1 or the interaction of PD-1 and PD-L1. The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier.

[0028] In a third aspect, the present application provides methods of treating cancer in a subject in need thereof, comprising administering to the subject i) an antibody, or antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or antigen-binding portion thereof, capable of inhibiting the binding of PD-1 to PD-L1 or the interaction of PD-1 and PD-L1. In some embodiments, the method comprises administering to the subject a composition of the present application.

[0029] The cancer may be a solid tumor or a hematological cancer, including, but not limited to, gastric cancer, liver cancer, colorectal cancer, non-small cell lung cancer, small cell lung cancer, ovarian cancer, pancreatic cancer, breast cancer, cervical cancer, endometrial cancer, head and neck tumors, urothelial carcinoma, bile duct cancer, bladder cancer, glioblastoma, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), multiple myeloma, Hodgkin's lymphoma (HL), and non-Hodgkin's lymphoma (NHL). In some embodiments, the cancer may be a cancer that expresses CD24, PD-L1, and / or PD-L2, or a cancer that does not express CD24, PD-L1, or PD-L2. In some embodiments, the cancer may be a cancer that highly expresses PD-L1. In some embodiments, the cancer may be a cancer with low PD-L1 expression. In some embodiments, the cancer may be colorectal cancer.

[0030] The present application also relates to the use of the present compositions in the treatment of cancer and in the preparation of a medicament for treating cancer.

[0031] Other features and advantages of the present disclosure will be apparent from the following detailed description and examples, which should not be construed as limiting. All references, Genbank entries, patents, and published patent applications cited throughout this application are hereby incorporated by reference. [Brief explanation of the drawings]

[0032] The following detailed description is by way of example and is not intended to limit the application to only the particular embodiments described, and can be best understood with reference to the accompanying drawings.

[0033] [Figure 1] The mean tumor size change in each group of mice treated with IMM47 and tislelizumab is shown.

[0034] [Figure 2] Tumor size changes in individual mice from each group treated with IMM47 and tislelizumab are shown.

[0035] [Figure 3] Shows the mean tumor size change in each group of mice treated with IMM47 in combination with Opdivo® and Keytruda®.

[0036] [Figure 4] Shows the change in tumor size for individual mice from each group treated with IMM47 in combination with Opdivo® and Keytruda®. DETAILED DESCRIPTION OF THE INVENTION

[0037] Before particular embodiments of the present application are disclosed and described, it is to be understood that this invention is not limited to the particular methods and materials disclosed herein, as these may vary to some extent. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not limiting, since the scope of the present application will be limited only by the appended claims and their equivalents.

[0038] As used herein, the term "antibody" includes whole antibodies, e.g., IgG, IgA, IgD, IgE, and IgM, as well as any antigen-binding fragment (or antigen-binding portion) or single chain thereof. Whole antibodies are glycoproteins comprising at least two heavy chains and two light chains, which are linked by disulfide bonds. Each heavy chain contains a heavy chain variable region (V H ) and the heavy chain constant region. The heavy chain constant region is made up of three domains, C H1 , C H2 , and C H3 Each light chain comprises a light chain variable region (V L ) and a light chain constant region. The light chain constant region comprises one domain, C L V H and V L The regions can be further subdivided into regions of hypervariability, called complementarity-determining regions, interspersed with more conserved regions, called framework regions (FR). H or VL consists of three CDRs and four FRs arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant region of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0039] The term "antigen-binding portion" of an antibody (or simply "antibody portion") refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody include (i) Fab fragments, V L , V H , C L and C H1 (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a V H and C H1 (iv) Fd fragments consisting of a single group of antibody V L and V H Fv fragment consisting of domains, (v) V H (vi) isolated complementarity determining regions (CDRs). L and V H Although the two domains of the Fv fragment are encoded by separate genes, they can be joined by a synthetic linker using recombinant methods, where the synthetic linker connects the V L and V HThese can be configured as a single protein chain (known as single-chain Fv (scFv)) in which the regions pair to form a monovalent molecule. Such single-chain antibodies are also intended to be encompassed by the term "antigen-binding portion" of an antibody. These antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies.

[0040] The heavy and light chain variable region CDRs of the antibodies, or antigen-binding portions thereof, of the present disclosure are defined according to the IMGT numbering system. As is well known in the art, the heavy and light chain variable region CDRs can be determined, for example, by the Chothia, Kabat, AbM, or Contact numbering systems / methods.

[0041] As used herein, the term "murine" heavy / light chain variable region refers to a variable region in which the framework (FR) and CDR regions are all derived from mouse germline immunoglobulin sequences, while "humanized" heavy / light chain variable region refers to a variable region derived from a non-human species but that has been modified to increase its similarity to antibody variants naturally occurring in humans.

[0042] The term "antibody or antigen-binding portion thereof capable of inhibiting the binding / interaction of PD-1 and PD-L1" refers to any antibody or antigen-binding portion thereof capable of blocking or suppressing the PD-1 signaling pathway by inhibiting the binding or interaction of PD-1 and PD-L1, and includes, but is not limited to, anti-PD-1 antibodies or antigen-binding portions thereof that specifically bind to PD-1 and inhibit PD-1-PD-L1 interaction, anti-PD-L1 antibodies or antigen-binding portions thereof that specifically bind to PD-L1 and inhibit PD-1-PD-L1 interaction, and anti-PD-L1 antibodies or antigen-binding portions thereof that specifically bind to PD-L1 and achieve PD-1-PD-L1 inhibition by inducing ADCC or the like and killing PD-L1 cells.

[0043] The term "antagonist" or "blocking" antibody or antigen-binding portion thereof refers to an antibody or antigen-binding portion thereof that specifically binds a particular antigen and blocks this antigen-mediated signaling pathway. For example, an antagonist anti-PD-1 antibody or antigen-binding portion thereof refers to an antibody or antigen-binding portion thereof that specifically binds PD-1 and blocks the PD-1 signaling pathway induced by the binding of, for example, PD-L1 or PD-L1 plus PD-L2 to PD-1; an antagonist anti-PD-L1 antibody or antigen-binding portion thereof refers to an antibody or antigen-binding portion thereof that specifically binds PD-L1 and blocks the PD-1 signaling pathway induced by PD-1-PD-L1 interaction, including anti-PD-L1 antibodies or antigen-binding portions thereof that specifically bind PD-L1 and inhibit PD-1-PD-L1 interaction, and anti-PD-L1 antibodies or antigen-binding portions thereof that specifically bind PD-L1 and achieve PD-1-PD-L1 blockade by inducing ADCC or the like and killing PD-L1 cells.

[0044] As used herein, "high expression" or "low expression" of PD-L1 may be determined according to criteria known in the art or the criteria described in Ozawa Y et al. (2021) Cancer Sci. 112(1):72-80. The cells used in the examples to construct mouse tumor models had "high expression" of PD-L1 relative to common PD-L1-expressing tumor cells.

[0045] The terms "antibody-dependent cytotoxicity," "antibody-dependent cell-mediated cytotoxicity," or "ADCC" refer to a cell-mediated immune defense in which effector cells of the immune system actively lyse target cells, such as cancer cells, whose membrane surface antigens are bound by antibodies, such as anti-CD24 antibodies.

[0046] As used herein, the term "sequence identity" refers to the percentage of nucleotides / amino acids in a sequence that are identical to the nucleotides / amino acid residues in a reference sequence after sequence alignment, where spaces are introduced into the sequence alignment, if necessary, to achieve the maximum percentage of sequence identity between the two sequences. Those skilled in the art can perform pairwise or multiple sequence alignments and determine the percentage of sequence identity between two or more nucleic acid or amino acid sequences by various methods, such as using computer software such as ClustalOmega, T-coffee, Kalign, MAFFT, etc.

[0047] The term "subject" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, cows, and horses, although mammals such as non-human primates, sheep, dogs, cats, cows, horses, chickens, amphibians, and reptiles are preferred.

[0048] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and / or dosage forms that, within the scope of lawful and medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic response, and / or other problem or complication, and that meet a reasonable benefit / risk ratio.

[0049] As used herein, the term "effective amount" refers to the amount of a drug or pharmaceutical agent, such as an antibody or antigen-binding portion thereof, or composition of the present application, that is sought by an investigator or clinician to elicit a biological or medical response, e.g., in a tissue, system, animal, or human. Additionally, the term "therapeutically effective amount" refers to any amount that results in an improved treatment, cure, prevention, or reduction of a disease, condition, or side effect, or a reduced rate of progression of a disease or condition, compared to a comparable subject who does not receive such amount. An effective amount may be administered in one or more administrations, applications, or dosages, and is not intended to be limited to a particular formulation or route of administration. For example, a "therapeutically effective amount" of a composition or antibody combination of the present application preferably results in a reduction in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods of the disease, or prevention of functional impairment or disability due to the affliction of the disease. For example, for the treatment of a cancer-bearing subject, a "therapeutically effective amount" inhibits tumor growth by at least about 40%, more preferably at least about 60%, more preferably at least about 80%, and more preferably at least about 99% relative to an untreated subject.

[0050] As used herein, the term "treatment" includes any effect, such as, for example, alleviation, reduction, modulation, amelioration, or elimination, that results in an improvement in, or amelioration of, a condition, illness, disease, and the like, or an improvement in its symptoms.

[0051] As used herein, the term "pharmaceutical composition" refers to a combination of an active agent and a carrier (inert or active) that makes the composition particularly suitable for in vivo or in vitro diagnostic or therapeutic use.

[0052] The present application provides a combination therapy for cancer, comprising administering to a subject: i) an antibody, or antigen-binding portion thereof, capable of specifically binding to CD24, and ii) an antibody, or antigen-binding portion thereof, capable of inhibiting the binding or interaction of PD-1 and PD-L1.

[0053] In one embodiment, the antibody or antigen-binding portion thereof capable of specifically binding to CD24 comprises a heavy chain variable region CDR1 (HV-CDR1), HV-CDR2, HV-CDR3, and a light chain variable region CDR1 (LV-CDR1), LV-CDR2, and LV-CDR3, which comprise the amino acid sequences GYSITSGYS (SEQ ID NO: 1), IHYSGST (SEQ ID NO: 2), ARGADYALDY (SEQ ID NO: 3), QSLLYSSNQKNY (SEQ ID NO: 4), WAS, and QQNFIYPLT (SEQ ID NO: 5), respectively. + It may contain an FcR-binding heavy chain constant region to induce antibody-dependent cell-mediated cytotoxicity (ADCC) against tumor cells. Exemplary antibodies, including IMM47, IMM47C, and IMM47H, are described in Chinese Patent Application No. CN202111195246.5 and target CD24 + their binding ability to cells, CD24 + It has been shown to possess the ability to induce ADCC against cells and potent in vivo antitumor activity.

[0054] An antibody or antigen-binding portion thereof capable of inhibiting the binding or interaction of PD-1 and PD-L1 refers to any antibody or antigen-binding portion thereof that is capable of blocking or inhibiting the PD-1 signaling pathway, including, but not limited to, an anti-PD-1 antibody or antigen-binding portion thereof that specifically binds to PD-1 and inhibits PD-1-PD-L1 interaction, an anti-PD-L1 antibody or antigen-binding portion thereof that specifically binds to PD-L1 and inhibits PD-1-PD-L1 interaction, and an anti-PD-L1 antibody that specifically binds to PD-L1 and achieves PD-1-PD-L1 inhibition by inducing ADCC or the like to kill PD-L1 cells.

[0055] It is well known in the field of antibodies that the CDR regions are important for the antigen-binding ability of an antibody, while amino acids in the framework regions of the variable region can be modified or changed to some extent without changing the antigen-binding ability and binding specificity of the antibody.

[0056] The combination therapy of the present application can be used to treat cancer, including, but not limited to, colorectal cancer. In some embodiments, the combination therapy of the present application has demonstrated synergistic effects with significantly better tumoricidal activity than targeting CD24 alone or targeting PD-1 / PD-L1 alone. Furthermore, the dose of each antibody can be reduced to some extent compared to single-targeted therapy, and the combination therapy further exhibits better therapeutic effects than single-targeted therapy.

[0057] The combination therapy of the present application can be applied to animals, preferably mammals (e.g., livestock, cats, dogs, mice, rats), and more preferably humans. Any method of administration can be used to deliver both of the antibodies of the present application to a subject in need thereof. In some embodiments, the two antibodies in the combination of the present application are administered parenterally, for example, via intraperitoneal injection, intravenous injection, and the like.

[0058] One or more other drugs or treatments may optionally be used in the combination therapy of the present application, such as other chemotherapeutic or other anti-cancer drugs, immunostimulatory drugs, immunosuppressants, anti-tumor vaccines, and / or cytokine therapy (e.g., IL2 and GM-CSF). The other drugs may be combined with the two antibodies of the present application in a single dosage form, or these therapeutic agents may be used as separate dosage forms.

[0059] The combination therapy of the present application can be administered simultaneously as a single composition in a pharmaceutically acceptable carrier, or simultaneously as separate compositions of each agent in a pharmaceutically acceptable carrier. When more than one dose of the combination therapy is administered sequentially, the order of sequential administration can be reversed at each time of administration or can remain the same. Sequential administration can be combined with simultaneous administration or any combination thereof. Compositions and Uses

[0060] The present application provides pharmaceutically acceptable compositions comprising a therapeutically effective amount of an antibody or antigen-binding portion thereof capable of specifically binding to CD24, formulated together with one or more pharmaceutically acceptable carriers, a therapeutically effective amount of an antibody or antigen-binding portion thereof capable of inhibiting the binding or interaction of PD-1 and PD-L1, formulated together with one or more pharmaceutically acceptable carriers, and, optionally, one or more other therapeutic agents, as desired.

[0061] Pharmaceutical compositions may include any number of carriers, including surfactants, thickening or emulsifying agents, solid binders, dispersing or suspending aids, solubilizing agents, colorants, flavoring agents, coatings, disintegrating agents, lubricants, sweeteners, preservatives, isotonic agents, and combinations thereof.

[0062] The pharmaceutical compositions are preferably suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active ingredient may be coated with a material to protect it from the action of acids and other natural conditions that may inactivate it. As used herein, the term "parenteral administration" refers to a method of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion. Alternatively, the compositions of the present disclosure can be administered via a parenteral route, such as a topical, epidermal, or mucosal route of administration, e.g., intranasal, oral, vaginal, rectal, sublingual, or topical.

[0063] The pharmaceutical compositions can be in the form of sterile aqueous solutions or dispersions, or they can be formulated as microemulsions, liposomes, or other ordered structures suitable to high drug concentration.

[0064] Dosage regimens for the pharmaceutical compositions of the present application vary depending on known factors such as the pharmacodynamic characteristics of each particular agent and its method and route of administration; the recipient's species, age, sex, health, condition, and weight; the nature and severity of symptoms; type of concurrent treatment; frequency of treatment; route of administration, the patient's renal and hepatic function, and the desired effect. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form varies depending on the subject being treated and the particular method of administration, and will generally be that amount of the composition that produces a therapeutic effect. Generally, out of 100 percent, this amount will range from about 0.01% to about 99% of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0065] Dosage regimens can be adjusted to provide the optimum desired response (e.g., therapeutic response). For example, a single bolus can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in unit dosage form for ease of administration and uniformity of dosing. As used herein, unit dosage form refers to physically discrete units suitable as a unitary dose for the subject to be treated; each unit contains a predetermined amount of active ingredient calculated to produce the desired therapeutic effect in combination with the necessary pharmaceutical carrier. Alternatively, antibodies can be administered as sustained-release formulations, requiring less frequent administration. Controlled-release formulations include implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. In certain embodiments, the compositions of the present disclosure can be formulated to ensure proper distribution in vivo. For example, to ensure that a composition of the present disclosure crosses the blood-brain barrier, it can be formulated in liposomes.

[0066] The present application also relates to in vivo gene therapy, in which nucleic acid molecules encoding two antibodies or antigen-binding portions thereof are directly introduced into a subject. For example, nucleic acid sequences encoding the antibodies or antigen-binding portions thereof of the present disclosure are introduced into target cells via local injection of a nucleic acid construct with or without an appropriate delivery vector, such as an adeno-associated viral vector. Alternative viral vectors include, but are not limited to, retrovirus, adenovirus, herpes simplex virus, and papillomavirus vectors. Physical introduction of a viral vector in vivo can be achieved by local injection of the desired nucleic acid construct or other appropriate delivery vector containing the desired nucleic acid sequence, liposome-mediated introduction, direct injection (naked DNA), or microparticle bombardment (gene gun).

[0067] The compositions of the present application can be used to treat cancer, including but not limited to, colorectal cancer. In some embodiments, administration of the compositions of the present application exhibits synergistic effects with significantly better tumoricidal activity compared to targeting CD24 alone and targeting PD-1 / PD-L1 alone.

[0068] The present application is further described with reference to the following non-limiting examples. example

[0069] IMM47 in the present application is an IgG antibody against CD24 and comprises two heavy chains and two light chains, wherein each heavy chain comprises a murine heavy chain variable region (SEQ ID NO: 6) and a human IgG1 constant region (SEQ ID NO: 8), and each light chain comprises a humanized light chain variable region (SEQ ID NO: 7) and a human K constant region (SEQ ID NO: 9).

[0070] IMM47 and two other anti-CD24 antibodies, IMM47C and IMM47H, which have the same heavy and light chain variable region CDRs, have similar CD24 activity. + Cell binding capacity, CD24 +These antibodies have demonstrated their ability to induce ADCC against human leukocytes and possess potent in vivo antitumor activity. In particular, IMM47C comprises a murine heavy chain variable region (SEQ ID NO: 6), a human IgG1 constant region (SEQ ID NO: 8), a murine light chain variable region (SEQ ID NO: 10), and a human K constant region (SEQ ID NO: 9); IMM47H comprises a humanized heavy chain variable region (SEQ ID NO: 11), a human IgG1 constant region (SEQ ID NO: 8), a humanized light chain variable region (SEQ ID NO: 12), and a human K constant region (SEQ ID NO: 9). More details can be found in Chinese Patent Application No. CN202111195246.5.

[0071] IMM2515H in the present application is an IgG antibody against PD-L1 comprising two heavy chains and two light chains, wherein each heavy chain comprises a heavy chain variable region (SEQ ID NO: 18) and a human IgG1 constant region (SEQ ID NO: 8), and each light chain comprises a light chain variable region (SEQ ID NO: 19) and a human K constant region (SEQ ID NO: 9). Example 1. In vivo antitumor effects of IMM47 in combination with tislelizumab

[0072] The human CD24 gene (Uniprot ID#P25063) was cloned into the neomycin-resistant expression vector pMac-G418 using the restriction enzyme cleavage sites Hind III and Not I to obtain the hCD24 expression plasmid. In addition, the human PD-L1 gene (Uniprot ID#Q9NZQ7) was cloned into the puromycin-resistant expression vector pMac-Pur using the restriction enzyme cleavage sites Hind III and Not I to obtain the hPD-L1 expression plasmid.

[0073] MC38 cells were transiently transfected with the above hCD24 expression plasmid using the drug PEI, and screened under neomycin selection pressure for multiple clones that stably express human CD24, which were then subcloned into 96-well plates using limiting dilution to select stable single-clonal cell lines with high CD24 expression. Using the same procedure as above, the selected MC38 cells that stably and highly express human CD24 were transfected with the above hPD-L1 expression plasmid and screened using puromycin for single-clonal cell lines that could stably and highly express both human CD24 and human PD-L1.

[0074] MC38-hCD24 / hPD-L1 cells prepared as described above were cultured in high-carbohydrate DMEM medium containing 10% inactivated fetal bovine serum in an incubator at 37°C under 5% CO2. Cells were separated into flasks and subcultured every 2 to 3 days when they reached 80% confluency.

[0075] MC38-hCD24 / hPD-L1 cells were harvested in log phase and resuspended in PBS for cell counting at a cell density of 3.0 × 10 7 The cell suspension was adjusted to approximately 3.0 × 10 cells / mL for each mouse. 6 The cells were subcutaneously inoculated into the right flank of 6-8 week old C57BL / 6-hPD-1 mice using a 1 mL syringe, 100 μL per mouse.

[0076] The average tumor size was approximately 89 mm 3 When tumor size reached 100 μg / kg, mice with medium-sized tumors were selected. Animals were randomly assigned to test groups according to tumor volume, with six animals in each group. The day of grouping was defined as D0, and intraperitoneal (ip) injections of the drug were initiated on that day. The specific dosing regimen is shown in Table 1. Table 1. Dosing regimens [Table 1]

[0077] Tumor size was measured twice a week and mice were observed until D33. Mice from the vehicle group had tumors of 3000 mm 3 The tumor volume (TV) was calculated as TV = 1 / 2 × a × b 2 Here, "a" represents the long diameter of the tumor, and "b" represents the short diameter of the tumor.

[0078] Relative tumor volume (RTV) is RTV=V t / V initial × 100%, where V initial represents the tumor size measured at the time of grouping (i.e., D0), and V t represents the tumor size at each measurement. The relative tumor growth rate T / C (%) is calculated as T / C (%) = (T RTV / C RTV ) × 100%, where T RTV represents the relative tumor volume in the treatment group, and C RTV represents the relative tumor volume in the vehicle group. The tumor volume inhibition rate (TGI) is calculated as follows: TGI = [1-(TV t -TV initial ) / (CV t -CV initial )] × 100%, where TV t represents the tumor volume of the treatment group at each measurement, and TV initial represents the tumor volume of the treatment group at the time of grouping, and CV t represents the tumor volume of the control group at each measurement, and CV initial represents the tumor volume of the control group at the time of grouping. The body weight change rate (BWC) of the animals was (BW final -BW initial ) / BW initial × 100%, where BWinitial represents the body weight of the animals at the time of grouping, and BW final represents the body weight of the animal at each measurement. The tumor weight inhibition rate (IR) was calculated as (W C -W T ) / W C × 100%, where W C represents the tumor weight in the control group, and W T indicates tumor weight in the treatment group. Raw data were measured and recorded, and analysis was performed based on the raw data. The analysis results were expressed as mean and standard error (Mean ± SEM), and the difference in tumor volume between the control group and the treatment group was analyzed using a t-test, with p<0.05 indicating a statistical difference.

[0079] Figure 1 shows that tislelizumab had some antitumor effects, with the best effects being achieved by administration of IMM47 alone and by the combination of IMM47 and tislelizumab. Figure 2 shows the changes in tumor volume in individual mice, demonstrating that the antitumor effect of simultaneous administration of IMM47 and tislelizumab was even better than that of administration of IMM47 alone. In particular, the complete remission (CR) rate in the IMM47 and tislelizumab simultaneous administration group reached 100%, while the CR rate in the IMM47 alone administration group was slightly lower.

[0080] It can be seen that IMM47 and tislelizumab act synergistically against tumors such as colon cancer, resulting in the complete elimination of tumor cells. Example 2. In vivo antitumor efficacy of IMM47 in combination with Opdivo® or Keytruda®

[0081] MC38-hCD24 / hPD-L1 cells were cultured in high-carbohydrate DMEM medium containing 10% inactivated fetal bovine serum in an incubator at 37°C under 5% CO2. Cells were dissociated into flasks and subcultured every 2 to 3 days when they reached 80% confluency.

[0082] MC38-hCD24 / hPD-L1 cells were harvested in log phase and resuspended in PBS for cell counting at a cell density of 3.0 × 10 7 The cell suspension was adjusted to approximately 3.0 × 10 cells / mL for each mouse. 6 The cells were subcutaneously inoculated into the right flank of 6-8 week old C57BL / 6-hPD-1 mice using a 1 mL syringe, 100 μL per mouse.

[0083] The average tumor size is approximately 90 mm 3 When tumor volume reached 100 μg / kg, mice with medium-sized tumors were selected. Animals were randomly assigned to test groups according to tumor volume, with six animals in each group. The day of grouping was defined as D0, on which intraperitoneal (ip) injections of drugs were initiated. The specific dosing regimen is shown in Table 2. Table 2. Dosing regimens [Table 2]

[0084] Tumor size was measured twice a week until D24. Tumor volume (TV) was calculated as TV = 1 / 2 × a × b 2 where "a" represents the long diameter of the tumor and "b" represents the short diameter of the tumor.

[0085] Relative tumor volume (RTV), relative tumor growth rate (T / C) (%), tumor volume inhibition rate (TGI), animal body weight change (BWC), and tumor weight inhibition rate (IR) were calculated as described above. Raw data were measured and recorded, and analyses were performed based on the raw data. The results were expressed as mean and standard error (mean ± SEM). The difference in tumor volume between the control group and the treatment group was analyzed using a t-test, with p<0.05 indicating a statistical difference.

[0086] It can be seen from Figure 3 that administration of IMM47 alone, administration of Opdivo® alone, and administration of Keytruda® alone were effective in suppressing tumor growth, while co-administration of IMM47 and Opdivo® and co-administration of IMM47 and Keytruda® resulted in complete elimination of tumor tissue. Furthermore, the changes in individual tumors in Figure 4 show that the complete response rate (CR) was extremely high in the IMM47-Opdivo® or -Keytruda® combination group, which was much higher than that in the other groups, indicating that the combination synergistically suppressed tumor growth.

[0087] Although administration of IMM47 alone at a high dose showed a strong antitumor effect, which may have masked to some extent the synergistic effect when IMM47 was used in combination with an anti-PD-1 antibody, the results of Examples 1 and 2 show that when the dose of IMM47 was reduced, IMM47 exerted a synergistic effect with the anti-PD-1 antibody. Therefore, the combination of IMM47 with an anti-PD-1 antibody can achieve excellent antitumor effects even at lower doses. Example 3. In vivo antitumor effect of IMM47 in combination with anti-PD-L1 antibody

[0088] In vivo antitumor efficacy was further measured for IMM47 in combination with Tecentriq® atezolizumab or the PD-L1 antibody IMM2515H.

[0089] MC38-hCD24 / hPD-L1 cells were cultured in high-carbohydrate DMEM medium containing 10% inactivated fetal bovine serum in an incubator at 37°C under 5% CO2. Cells were dissociated into flasks and subcultured every 2 to 3 days when they reached 80% confluency.

[0090] MC38-hCD24 / hPD-L1 cells were harvested in log phase and resuspended in PBS for cell counting at a cell density of 3.0 × 10 7 The cell suspension was adjusted to approximately 3.0 × 10 cells / mL for each mouse. 6The cells were subcutaneously inoculated into the right flank of 6-8 week old C57BL / 6-hPD-1 mice using a 1 mL syringe, 100 μL per mouse.

[0091] The average tumor size is approximately 100 mm 3 When tumor volume reached 100 μg / kg, mice with medium-sized tumors were selected. Animals were randomly assigned to test groups according to tumor volume, with six animals in each group. The day of grouping was defined as D0, and intraperitoneal (ip) injection of drugs into mice was initiated on that day. The specific dosing regimen is shown in Table 3. Table 3. Dosing regimens [Table 3]

[0092] Tumor size was measured twice a week until D24. Tumor volume (TV) was calculated as TV = 1 / 2 × a × b 2 where "a" represents the long diameter of the tumor and "b" represents the short diameter of the tumor.

[0093] Relative tumor volume (RTV), relative tumor growth rate (T / C) (%), tumor volume inhibition rate (TGI), animal body weight change (BWC), and tumor weight inhibition rate (IR) were calculated as described above. Raw data were measured and recorded, and analyses were performed based on the raw data. The results were expressed as mean and standard error (mean ± SEM). The difference in tumor volume between the control group and the treatment group was analyzed using a t-test, with p<0.05 indicating a statistical difference.

[0094] The sequence information in this application is summarized as follows: [Table 4] JPEG2025540021000005.jpg149130

[0095] While the present disclosure has been described above with reference to one or more embodiments, it should be understood that the disclosure is not limited to those embodiments. The description in this disclosure is intended to include all modifications and equivalents as may be included within the spirit and scope of the appended claims. All references cited herein are incorporated by reference in their entirety.

Claims

1. i) an antibody or antigen-binding portion thereof capable of specifically binding to CD24, and ii) An antibody or antigen-binding portion thereof capable of inhibiting the binding of PD-1 and PD-L1 or the interaction between PD-1 and PD-L1. Equipped with the antibody or the antigen-binding portion thereof capable of specifically binding to CD24 has a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising HV-CDR1, HV-CDR2, and HV-CDR3, the light chain variable region comprising LV-CDR1, LV-CDR2, and LV-CDR3, and the HV-CDR1, the HV-CDR2, the HV-CDR3, the LV-CDR1, the LV-CDR2, and the LV-CDR3 comprise the amino acid sequences GYSITSGYS (SEQ ID NO: 1), IHYSGST (SEQ ID NO: 2), ARGADYALDY (SEQ ID NO: 3), QSLLYSSNQKNY (SEQ ID NO: 4), WAS, and QQNFIYPLT (SEQ ID NO: 5), respectively; composition.

2. 2. The composition of claim 1, wherein the heavy chain variable region and the light chain variable region of the antibody or the antigen-binding portion thereof, which can specifically bind to CD24, comprise the amino acid sequences of 1) SEQ ID NOs: 6 and 7, respectively; 2) SEQ ID NOs: 6 and 10, respectively; or 3) SEQ ID NOs: 11 and 12, respectively.

3. The composition of claim 1 , wherein the antibody or the antigen-binding portion thereof capable of specifically binding to CD24 further comprises a heavy chain constant region with FcR binding ability.

4. The composition according to claim 3 , wherein the heavy chain constant region having FcR binding ability is a human IgG1 heavy chain constant region.

5. 2. The composition of claim 1, wherein the antibody or the antigen-binding portion thereof capable of specifically binding to CD24 has a heavy chain constant region and a light chain constant region, wherein the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 8 and the light chain constant region comprises the amino acid sequence of SEQ ID NO:

9.

6. the antibody or the antigen-binding portion thereof capable of inhibiting binding of PD-1 to PD-L1 or PD-1-PD-L1 interaction; an antagonist anti-PD-1 antibody or antigen-binding portion thereof capable of specifically binding to PD-1; Antagonist anti-PD-L1 antibodies or antigen-binding portions thereof capable of specifically binding to PD-L1 2. The composition of claim 1, wherein:

7. 7. The composition of claim 6, wherein the antagonist anti-PD-1 antibody is selected from the group consisting of tislelizumab, nivolumab, and pembrolizumab.

8. the antagonist anti-PD-L1 antibody is selected from the group consisting of atezolizumab and IMM2515H; IMM2515H has a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising HV-CDR1, HV-CDR2, and HV-CDR3, and the light chain variable region comprising LV-CDR1, LV-CDR2, and LV-CDR3, and the HV-CDR1, the HV-CDR2, the HV-CDR3, the LV-CDR1, the LV-CDR2, and the LV-CDR3 comprise the amino acid sequences GYTFTSNW (SEQ ID NO: 13), IHPNSGSS (SEQ ID NO: 14), ARSYYGSSPYYFDY (SEQ ID NO: 15), QDIINY (SEQ ID NO: 16), YTS, and QQGDTLPWT (SEQ ID NO: 17), respectively. The composition of claim 6.

9. Use of an antibody or antigen-binding portion thereof capable of specifically binding to CD24, and an antibody or antigen-binding portion thereof capable of inhibiting the binding of PD-1 and PD-L1 or the interaction of PD-1 and PD-L1, in the preparation of a medicament for the treatment of cancer, wherein the antibody or the antigen-binding portion thereof capable of specifically binding to CD24 has a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising HV-CDR1, HV-CDR2, and HV-CDR3, and the light chain variable region comprises a heavy chain variable region and a light chain variable region. The variable regions include LV-CDR1, LV-CDR2, and LV-CDR3, and the LV-CDR1, the LV-CDR2, the LV-CDR3, the LV-CDR1, the LV-CDR2, and the LV-CDR3 comprise the amino acid sequences GYSITSGYS (SEQ ID NO: 1), IHYSGST (SEQ ID NO: 2), ARGADYALDY (SEQ ID NO: 3), QSLLYSSNQKNY (SEQ ID NO: 4), WAS, and QQNFIYPLT (SEQ ID NO: 5), respectively; use.

10. The use of claim 9, wherein the heavy chain variable region and the light chain variable region of the antibody or the antigen-binding portion thereof, which are capable of specifically binding to CD24, comprise the amino acid sequences of 1) SEQ ID NOs: 6 and 7, respectively; 2) SEQ ID NOs: 6 and 10, respectively; or 3) SEQ ID NOs: 11 and 12, respectively.

11. The use according to claim 9, wherein the antibody or the antigen-binding portion thereof capable of specifically binding to CD24 further comprises a heavy chain constant region with FcR binding ability.

12. The use of claim 9, wherein the antibody or the antigen-binding portion thereof capable of specifically binding to CD24 has a heavy chain constant region and a light chain constant region, wherein the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 8 and the light chain constant region comprises the amino acid sequence of SEQ ID NO:

9.

13. the antibody or the antigen-binding portion thereof capable of inhibiting binding of PD-1 to PD-L1 or PD-1-PD-L1 interaction; an antagonist anti-PD-1 antibody or antigen-binding portion thereof capable of specifically binding to PD-1; Antagonist anti-PD-L1 antibodies or antigen-binding portions thereof capable of specifically binding to PD-L1 The use according to claim 9, wherein

14. 14. The use of claim 13, wherein the antagonist anti-PD-1 antibody is selected from the group consisting of tislelizumab, nivolumab, and pembrolizumab.

15. the antagonist anti-PD-L1 antibody is selected from the group consisting of atezolizumab and IMM2515H; IMM2515H has a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising HV-CDR1, HV-CDR2, and HV-CDR3, and the light chain variable region comprising LV-CDR1, LV-CDR2, and LV-CDR3, and the HV-CDR1, the HV-CDR2, the HV-CDR3, the LV-CDR1, the LV-CDR2, and the LV-CDR3 comprise the amino acid sequences GYTFTSNW (SEQ ID NO: 13), IHPNSGSS (SEQ ID NO: 14), ARSYYGSSPYYFDY (SEQ ID NO: 15), QDIINY (SEQ ID NO: 16), YTS, and QQGDTLPWT (SEQ ID NO: 17), respectively.

14. The use according to claim 13.

16. 10. The use of claim 9, wherein the cancer is selected from the group consisting of gastric cancer, liver cancer, colorectal cancer, non-small cell lung cancer, small cell lung cancer, ovarian cancer, pancreatic cancer, breast cancer, cervical cancer, endometrial cancer, head and neck tumors, urothelial carcinoma, bile duct cancer, bladder cancer, glioblastoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), multiple myeloma, Hodgkin's lymphoma (HL), and non-Hodgkin's lymphoma (NHL).

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