Method for treatment of advanced or metastatic solid cancer using Anti-CD47 antibody

Abstract

Provided is a method for treating solid cancer using an anti-CD47 antibody.

Description

Treatment of advanced or metastatic solid tumors using anti-CD47 antibodies

[0001] This patent application claims priority to Republic of Korea Patent Application No. 10-2024-0066776, filed with the Korean Intellectual Property Office on May 22, 2024, the disclosure of which is incorporated herein by reference.

[0002] The present invention relates to a method for treating patients with advanced or metastatic cancer using an anti-CD47 antibody.

[0003] Sequence list

[0004] This application includes a sequence listing, which has been submitted electronically in XML format and is incorporated herein by reference in its entirety. A copy of said sequence listing, dated October 15, 2024, is named KC24163.xml and is 10.3 kilobytes in size.

[0005] CD47 is a transmembrane protein expressed on the surface of cancer cells that transmits a "don't-eat-me" signal to signal-regulatory protein alpha (SIRPα), a receptor on phagocytes such as macrophages and dendritic cells. Binding of CD47 to SIRPα induces phosphorylation of the immunoreceptor tyrosine-based inhibitory motif (IBM) in the cytoplasmic domain of SIRPα, which then recruits Src homologous phosphatases 1 and 2 (SHP-1 and SHP-2), thereby inhibiting phagocyte signaling (Oldenborg 2013). This CD47 inhibitory mechanism is frequently overexpressed in various cancer cells, including hematological malignancies and solid tumors. In various animal models, it has been confirmed that blocking CD47 / SIRPα induces macrophage phagocytosis of cancer cells, promoting antitumor activity (Tseng D, et al. 2013; Majeti R, et al. 2009). In particular, CD47 blockade induces antitumor T cell responses by cross-presenting tumor antigens after phagocytosis of cancer cells by dendritic cells (Liu X, et al. 2015). In addition, combining CD47 / SIRPα with chemotherapy or immunomodulatory drugs such as programmed cell death protein 1 (PD-1) and programmed death ligand 1 (PD-L1) enhances anticancer effects (Gordon SR, et al. 2017; Liu X, et al. 2018; Lian S, et al. 2019).

[0006] Currently, various anticancer drugs targeting CD47 / SIRPα are in preclinical or clinical development. According to the related website (Clinicaltrials.gov), 12 drugs are currently in development as anticancer agents. Magrolimab (Hu5F9-G4, an anti-CD47 monoclonal antibody) has been reported to be effective as a monotherapy in patients with acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and some solid tumors (clear cell ovarian cancer and fallopian tube carcinoma). However, due to serious safety issues, the drug recently announced a halt to clinical trials for hematological malignancies and a suspension of clinical trial subject recruitment for solid tumors. Therefore, there is a critical need for anticancer drugs with safety, tolerability, and efficacy, especially for patients with advanced or metastatic cancer who have no approved or established therapeutic alternatives.

[0007] IMC-002, an anti-CD47 antibody, is a human IgG4 antibody with a hinge-stabilizing S228P mutation that prevents Fab substitution. IMC-002 targets CD47 and inhibits the interaction between CD47 and SIRPα, thereby enhancing macrophage phagocytosis of cancer cells in vitro and inhibiting tumor growth in vivo (in a xenograft model). IMC-002 does not bind well to red blood cells, and thus did not cause hemagglutination in vitro or anemia in vivo. These results suggest the potential of IMC-002 as a safe, well-tolerated, and effective anticancer agent.

[0008] [Prior Art Literature]

[0009] [Patent Document]

[0010] (Patent Document 0001) Korean Patent Publication No. 10-2625835

[0011] [Non-patent literature]

[0012] (Non-patent document 0001) Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242, 1991

[0013] (Non-patent literature 0002) Lefranc et al, Dev. Comp. Immunol. 29:185-203; 2005

[0014] (Non-patent literature 0003) Honegger and Pluckthun, J. Mol. Biol. 309(3):657-670; 2001

[0015] (Non-patent literature 0004) Tseng D, et al. 2013, Majeti R, et al. 2009

[0016] (Non-patent literature 0005) Gordon SR, et al. 2017 Liu X, et al. 2018, Lian S, et al. 2019

[0017] The purpose of the present invention is to solve all of the problems of the above-mentioned prior art.

[0018] The present invention aims to provide a method, pharmaceutical composition and kit for administering an anti-CD47 antibody to a patient with a solid cancer to treat the tumor or inhibit tumor proliferation.

[0019] The purpose of the present invention is not limited to the purposes mentioned above. The purpose of the present invention will become more apparent from the following description and may be realized by the means and combinations thereof set forth in the claims.

[0020] A representative configuration of the present invention to achieve the above purpose is as follows.

[0021] One aspect of the present invention provides a pharmaceutical composition for treating a tumor or inhibiting tumor growth in a patient with advanced or metastatic solid tumor, comprising an effective amount of an anti-CD47 antibody that specifically binds to CD47 and comprises HCDR1, HCDR2 and HCDR3 contained in a heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in a light chain variable region (LCVR) of SEQ ID NO: 2, wherein the antibody is administered to the patient at a dose of about 5 mg / kg to about 30 mg / kg every two to three weeks without priming.

[0022] One aspect of the present invention provides a method for treating a tumor or inhibiting tumor growth, comprising the steps of: (a) selecting a patient with an advanced or metastatic solid tumor; and (b) administering to the selected patient an anti-CD47 antibody or a pharmaceutical composition disclosed herein at a dose of about 5 mg / kg to about 30 mg / kg without priming every two to about three weeks, wherein the antibody is an anti-CD47 antibody that specifically binds to CD47 and comprises HCDR1, HCDR2 and HCDR3 contained in a heavy chain variable region of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in a light chain variable region of SEQ ID NO: 2.

[0023] Another aspect of the present invention provides a kit comprising an anti-CD47 antibody together with instructions for use of the anti-CD47 antibody for treating a tumor or inhibiting tumor growth in a patient with a solid tumor, wherein the instructions for use include instructions for a method of treating or inhibiting the tumor as disclosed herein, comprising administering the antibody to the patient without priming at a dose of about 5 mg / kg to about 30 mg / kg every two to about three weeks, wherein the antibody specifically binds to CD47 and comprises HCDR1, HCDR2 and HCDR3 contained in a heavy chain variable region of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in a light chain variable region of SEQ ID NO: 2.

[0024] In some embodiments, the antibody may be a fully human anti-CD47 antibody (e.g., the IMC-002 antibody).

[0025] In some embodiments, the heavy chain variable region of the antibody may comprise HCDR1 of SEQ ID NO: 5, HCDR2 of SEQ ID NO: 6, and HCDR3 of SEQ ID NO: 7, and the light chain variable region may comprise LCDR1 of SEQ ID NO: 8, LCDR2 of SEQ ID NO: 9, and LCDR3 of SEQ ID NO: 10.

[0026] In some embodiments, the heavy chain variable region of the antibody may comprise the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 95% identical thereto, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO: 2 or an amino acid sequence at least 95% identical thereto.

[0027] In some embodiments, the antibody may not cause significant levels of red blood cell aggregation or anemia in the patient.

[0028] In some embodiments, the composition comprises about 10 to 30 mM sodium phosphate, about 80 to 120 mM sodium chloride, about 50 to 80 mM mannitol, and about 0.01 to 0.1% polysorbate 80, and may have a pH of about 4 to 7.

[0029] In some embodiments, the composition may comprise about 20 mM sodium phosphate, about 100 mM sodium chloride, about 65.9 mM mannitol, and 0.05% polysorbate 80 and have a pH of about 5.5.

[0030] In some embodiments, the composition may be in a liquid dosage form for intravenous injection or infusion.

[0031] In some embodiments, the solid tumor may be liver cancer (e.g., hepatocellular carcinoma), breast cancer (e.g., triple negative breast cancer), or gallbladder cancer.

[0032] In some embodiments, the patient may be one who is ineligible for standard treatment or has failed standard treatment.

[0033] In some embodiments, the antibody may be administered to the patient about every two weeks or about every three weeks.

[0034] In some embodiments, the antibody may be administered to the patient at a dose of about 20 mg / kg or about 30 mg / kg.

[0035] In some embodiments, the patient may have received at least one prior systemic treatment prior to administration of the anti-CD47 antibody.

[0036] In some embodiments, the patient may be a stage 4 patient with multiple distant metastases or a locally advanced patient.

[0037] In some embodiments, the antibody may be administered in combination with other anticancer agents.

[0038] In some embodiments, the antibody may be administered in combination with lenvatinib.

[0039] In some embodiments, the antibody may be administered in combination with paclitaxel.

[0040] In some embodiments, the antibody may be administered in combination with gemcitabine, carboplatin, or a combination thereof.

[0041] In some embodiments, the antibody may be administered concurrently with, sequentially with, or in reverse order to other anticancer agents, with time intervals as necessary for treatment.

[0042] In some embodiments, lenvatinib may be administered daily.

[0043] In some embodiments, paclitaxel may be administered on day 1 of a cycle of antibody administration.

[0044] In some embodiments, gemcitabine and carboplatin may be administered on days 1 and 8 of the antibody dosing cycle.

[0045] In some embodiments, the treatment or inhibition method may further comprise measuring CD47 positive macrophage density in a biological sample obtained from the patient before or after administration of the anti-CD47 antibody.

[0046] In one aspect of the present invention, a pharmaceutical composition for treating a tumor or inhibiting tumor proliferation in a patient with advanced or metastatic hepatocellular carcinoma, comprising an effective amount of an IMC-002 antibody, wherein the antibody is administered to the patient at a dose of about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, or about 30 mg / kg about every two weeks or about three weeks without priming, is provided.

[0047] In some embodiments, a pharmaceutical composition for treating a tumor or inhibiting tumor growth in a patient with advanced or metastatic hepatocellular carcinoma may be administered to the patient in combination with levatinib.

[0048] In one aspect of the present invention, a pharmaceutical composition for treating a tumor or inhibiting tumor growth in a patient with advanced or metastatic gallbladder cancer, comprising an effective amount of an IMC-002 antibody, wherein the antibody is administered to the patient at a dose of about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, or about 30 mg / kg about every two weeks or about three weeks without priming is provided.

[0049] In one aspect of the present invention, a pharmaceutical composition for treating a tumor or inhibiting tumor growth in a patient with advanced or metastatic breast cancer, comprising an effective amount of an IMC-002 antibody, wherein the antibody is administered to the patient at a dose of about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, or about 30 mg / kg about every two weeks or about three weeks without priming is provided.

[0050] In some embodiments, the pharmaceutical composition for treating a tumor or inhibiting tumor growth in a patient with advanced or metastatic breast cancer may be administered to the patient in combination with a combination of paclitaxel or gemcitabine and carboplatin.

[0051] The anti-CD47 antibody IMC-002 induces macrophage phagocytosis of cancer cells and potently inhibits tumor growth in patients with advanced or metastatic cancer. IMC-002 was administered at a dose of approximately 5 mg / kg to approximately 30 mg / kg every two to three weeks without priming, demonstrating an excellent safety profile and efficacy in patients with advanced or metastatic solid tumors who had failed standard treatment.

[0052] Figure 1 shows the mechanism of action of an anti-CD47 antibody (IMC-002) according to one embodiment of the present invention.

[0053] Figure 2 shows a schematic diagram of a dose-escalation study and an expansion study according to a clinical trial design of one embodiment of the present invention.

[0054] Figure 3 shows the change in hemoglobin level according to the dose of IMC-002 administered in a dose-escalation study as a result of a clinical trial according to one embodiment of the present invention.

[0055] Figure 4 shows the change in neutrophil count according to the dose of IMC-002 administered in a dose-escalation study as a result of a clinical trial according to one embodiment of the present invention.

[0056] Figure 5 shows the change in platelet count according to the dose of IMC-002 administered in a dose-escalation study as a result of a clinical trial according to one embodiment of the present invention.

[0057] Figure 6 shows a swimmer plot of tumor response over time in a dose-escalation study as a result of a clinical trial according to one embodiment of the present invention. In the swimmer plot, the symbol ● indicates SD (Stable Disease), PD (Progressive Disease, progressive lesion), means continuing treatment.

[0058] Figure 7 shows the change rate of tumor burden of target lesions in a dose-escalation study as a result of a clinical trial according to one embodiment of the present invention.

[0059] FIG. 8A and FIG. 8B show the CD47 positive macrophage density and CD47 negative macrophage density in patients who achieved clinical benefit (CBR) and patients who did not achieve clinical benefit (Non-CBR) as a result of a clinical trial according to one embodiment of the present invention.

[0060] Figure 9 shows the simulated pharmacokinetic (PK) profile of IMC-002 after intravenous infusion every three weeks as a result of a clinical trial according to one embodiment of the present invention.

[0061] Figure 10 shows the results of a clinical trial according to one embodiment of the present invention, showing comparative results of PK profiles according to the administered dose.

[0062] Figure 11 shows the PK serum concentration after initial administration as a result of a clinical trial according to one embodiment of the present invention.

[0063] Figure 12 shows the change in hemoglobin level according to the dose of IMC-002 administered in an expansion study as a result of a clinical trial according to one embodiment of the present invention.

[0064] Figure 13 shows the change in neutrophil count according to the dose of IMC-002 administered in an expansion study as a result of a clinical trial according to one embodiment of the present invention.

[0065] Figure 14 shows the change in platelet count according to the dose of IMC-002 administered in an expansion study as a result of a clinical trial according to one embodiment of the present invention.

[0066] Figure 15 shows a swimmer plot of tumor response over time in an extension study as a result of a clinical trial according to one embodiment of the present invention. In the swimmer plot, the symbol ▲ indicates PR (Partial Response), ● indicates SD (Stable Disease), PD (Progressive Disease, progressive lesion), means continuing treatment.

[0067] Figure 16 shows the change rate of tumor burden of target lesions in an extended study as a result of a clinical trial according to one embodiment of the present invention.

[0068] The detailed description of the present invention, which follows, will be described with reference to specific drawings (if any) regarding specific embodiments in which the present invention may be practiced; however, the present invention is not limited thereto, but is defined solely by the appended claims to the full scope equivalent to or equivalent to what the claims describe. It should be understood that the various embodiments / embodiments of the present invention, while different from each other, are not necessarily mutually exclusive. For example, specific shapes, structures, and characteristics described herein may be modified from one embodiment / embodiment to another, or multiple embodiments / embodiments may be combined, without departing from the spirit and scope of the present invention. Technical and scientific terms used herein have the same meaning as commonly used in the art to which the present invention belongs, unless otherwise defined. In case of conflict, the present specification, including its definitions, will control. The definitions of terms set forth in this specification will be applied for the purpose of interpreting this specification, and terms expressed in the singular will be construed to also refer to the plural (i.e., at least one) and vice versa, unless the context otherwise makes it inappropriate.

[0069] definition

[0070] The term "about" as used herein refers to the typical error range for each value known to those of ordinary skill in the art. Furthermore, unless otherwise specified, all numbers, values, and / or expressions expressing ingredients, conditions, compositions, amounts, and so forth used herein should be understood to be modified by the term "about" because such numbers are approximations that inherently reflect, among other things, the various uncertainties of measurement that arise in obtaining such values. As examples, this may include ±10% or less, ±9% or less, ±8% or less, ±7% or less, ±6% or less, ±5% or less, ±4% or less, ±3% or less, ±2% or less, ±1% or less, or ±0.5% or less of a given numerical value.

[0071] The term "cancer" refers to a variety of diseases characterized by the uncontrolled growth of abnormal cells in the body. "Cancer" or "cancer tissue" may include tumors.

[0072] The term "advanced" is used in relation to cancer to refer to cancer that has not been completely removed or controlled by surgery, and is locally advanced. "Advanced cancer" is cancer that has reached at least stage IIIB.

[0073] The term "metastatic" is used in relation to cancer to refer to a cancer condition in which cancer cells break away from their original site of origin and travel, for example, through the blood or lymphatic system, to form tumors (metastatic tumors) elsewhere in the body. "Metastatic cancer" is cancer that has reached at least stage IV.

[0074] The term "standard treatment" refers to a treatment method generally recognized by medical professionals as appropriate for the treatment of a specific cancer, preferably a specific solid tumor. "Standard treatment" may be the same or different for different tumor types. "Standard treatment" may include first-line or second-line treatments, such as surgery, radiation therapy, chemotherapy, and immunotherapy. "Standard treatment" may be approved by various regulatory agencies, such as the U.S. Food and Drug Administration.

[0075] The term “antibody” includes monoclonal antibodies (including full-length antibodies) of any isotype, such as IgG, IgM, IgA, IgD, and IgE, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), antibody fusions (e.g., fusions of an antibody and a (poly)peptide, or of an antibody and a compound), and antibody fragments (including antigen-binding fragments). As used herein, the prefix “anti-”, when associated with an antigen, means that the antibody is reactive with that antigen. Antibodies reactive with a particular antigen can be produced by synthetic and / or recombinant methods, such as, but not limited to, screening of recombinant antibody libraries in phage or similar vectors, or by immunizing an animal with the antigen or an antigen-encoding nucleic acid. A representative IgG antibody is composed of two identical heavy chains and two identical light chains joined by disulfide bonds. Each heavy and light chain comprises a constant region and a variable region. The heavy chain variable region (HVR) and light chain variable region (LVR) each contain three segments, called “complementarity determining regions” (“CDRs”) or “hypervariable regions,” which are primarily involved in binding to antigenic epitopes. These are numbered sequentially from the N-terminus and are commonly referred to as CDR1, CDR2, and CDR3. The more conserved regions of the variable regions outside the CDRs are referred to as “framework regions” (“FRs”). As used herein, an antibody may be, for example, an animal antibody, a chimeric antibody, a humanized antibody, or a human antibody.

[0076] The term "monoclonal antibody" or "mAb" refers to a population of substantially homogeneous antibodies, the antibody molecules comprising the population being identical in amino acid sequence except for possible naturally occurring mutations that may be present in trace amounts. In contrast, conventional (polyclonal) antibody preparations typically comprise a plurality of different antibodies having different amino acid sequences in the variable domains, particularly CDR sequences, that are specific for different epitopes. "Monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in the therapeutic methods, medicaments, and uses disclosed may be produced by the hybridoma method first described by Kohler et al. (1975) Nature 256: 495, or by recombinant deoxyribonucleic acid (DNA) methods. "Monoclonal antibody" may also be produced by methods described, for example, by Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0077] The term "priming" refers to the initial antigenic stimulation administered prior to the start of treatment. "Priming" is used to activate the immune system or maximize the effectiveness of a treatment. For example, in certain immunotherapies, "priming" administration activates T cells, making subsequent treatments more effective. "Priming" refers to the primary antigenic stimulation, which induces an immune response to a specific antigen and then recalls a high level of immune response to that antigen upon subsequent reimmunization with the same antigen.

[0078] The term "pharmaceutical composition" or "pharmaceutical formulation" refers to a combination of an active ingredient with an inert or active carrier and / or excipient.

[0079] The term "effective amount" refers to an amount of a compound (including macromolecules such as antibodies) or composition (e.g., a compound or composition of the present invention) sufficient to achieve a beneficial or desired result. An effective amount may be administered in one or more doses, applications, or dosages, and is not intended to be limited to a particular formulation or route of administration.

[0080] The term "patient," "subject," "subject," or "subject" refers to a single subject who requires a therapy or treatment method, or who is participating in a clinical trial, epidemiological study, or used as a control, and includes human and mammalian veterinary patients, such as cattle, horses, dogs, and cats.

[0081] The term "administration" refers to the physical introduction of a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those of ordinary skill in the art. Exemplary routes of administration include oral, intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes. Parenteral routes of administration can be, for example, injection or infusion (e.g., intravenous infusion). For oral administration, the active ingredient may be formulated with pharmaceutically acceptable excipients in the form of tablets, capsules, sachets, tablets, etc. The term "parenteral administration" as used herein generally means any mode of administration other than enteral and topical, generally by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. Therapeutic agents may be administered parenterally or orally. Other parenteral routes include topical, epidermal, or mucosal administration, such as intranasal, vaginal, rectal, sublingual, or topical. Administration may be administered once, multiple times, and / or over an extended period of one day or more.

[0082] The term "combination administration" refers to the administration of two or more therapeutic agents. These types of therapeutic agents may be administered simultaneously, sequentially, or separately to a subject. Simultaneous administration refers to administering each type of therapeutic agent at once using the same or different administration methods, while sequential administration refers to administering each type of therapeutic agent separately and consecutively at regular intervals, with the time required between administrations being as short as possible. Separate administration refers to administering each type of therapeutic agent at regular intervals. These administration methods can be appropriately selected by a person skilled in the art, taking into account the patient's therapeutic efficacy and side effects. The frequency of administration can also be appropriately selected, taking into account the patient's condition and therapeutic efficacy, along with the administered dose. For example, it can be administered once or multiple times, and can be administered over one or more extended periods.

[0083] The term "biological sample" includes a clinical sample, and also includes tissue obtained by surgical resection, tissue obtained by biopsy, cultured cells, cell supernatants, cell lysates, tissue samples, organs, bone marrow, blood, plasma, serum, etc. A "biological sample" includes a sample suspected of containing target cells or normal control cells or such cells or biological fluids derived therefrom (e.g., cancerous cells, infected cells, etc.), for example, a sample containing polynucleotides and / or polypeptides obtained from such cells (e.g., a cell lysate or other cell extract containing polynucleotides and / or polypeptides). A biological sample comprising cells that are responsible for a patient's disease may also include cells that are not responsible for the disease.

[0084] The terms "phagocytic cell" and "phagocyte" are used interchangeably herein to refer to cells capable of phagocytosis. There are three main categories of phagocytes: macrophages, mononuclear cells (histocytes and monocytes); polymorphonuclear leukocytes (neutrophils), and dendritic cells.

[0085] Treatment methods

[0086] In one aspect of the present invention, a method of treating a patient with an effective amount of an anti-CD47 agent is provided. The invention is based, in part, on the clinical demonstration that both safety and efficacy can be achieved without priming when an anti-CD47 antibody (particularly IMC-002) is administered to patients with advanced or metastatic solid tumors at specific doses and cycles.

[0087] In some embodiments, the cancer may be an advanced or metastatic solid tumor. Specifically, the solid tumor may be one or more selected from lung cancer, pancreatic cancer, colon cancer, colorectal cancer, gallbladder cancer, bile duct cancer, stomach cancer, liver cancer, brain cancer, breast cancer, thyroid cancer, bladder cancer, esophageal cancer, ovarian cancer, melanoma, head and neck cancer, skin cancer, prostate cancer, hepatocellular carcinoma, fibrosarcoma, and cervical cancer. More specifically, the solid tumor may be liver cancer (e.g., hepatocellular carcinoma), breast cancer (e.g., triple-negative breast cancer), or gallbladder cancer.

[0088] In some embodiments, the patient may be ineligible for or have failed standard treatment. Specifically, standard treatment may be a treatment method generally recognized by medical professionals as appropriate for the treatment of a specific cancer, preferably a specific solid tumor. Specifically, standard treatment may include first-line or second-line treatment options, such as surgery, radiation therapy, chemotherapy, immunotherapy, or a combination thereof. Failure to standard treatment may mean relapse / refractory / progression after standard treatment.

[0089] In some embodiments, the patient may be a Stage 4 patient with multiple distant metastases or a locally advanced patient. A Stage 4 patient with multiple distant metastases or a locally advanced patient refers to, for example, a patient with histologically or cytologically proven metastatic or locally advanced liver cancer (e.g., hepatocellular carcinoma), breast cancer (e.g., triple-negative breast cancer), or gallbladder cancer.

[0090] In some embodiments, the methods of the present invention do not include a priming step with an anti-CD47 agent. Specifically, the absence of a priming step means that the preparatory step for activating the immune system in the early stages of cancer treatment is omitted. Omitting the priming step can simplify the treatment process, and may be particularly beneficial for elderly patients or those with other health problems. Omitting the priming step can also reduce the side effects of cancer treatment.

[0091] In some embodiments, the anti-CD47 agent of the present invention may be an anti-CD47 antibody, specifically a fully human anti-CD47 monoclonal antibody. A fully human antibody of the present invention is an antibody that has an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human and / or made using any technique for making human antibodies.

[0092] In some embodiments, the anti-CD47 antibody of the present invention is an IgG4 antibody. Specifically, the anti-CD47 antibody is an IgG4 kappa isotype antibody. More specifically, the anti-CD47 antibody is composed of two identical gamma heavy chains and two identical kappa light chains, and is formed into a glycosylated tetramer linked by disulfide bonds.

[0093] In some embodiments, the anti-CD47 antibody of the invention comprises a hinge region mutation, S228P, which prevents Fab substitution. The S228P mutation reduces Fab arm exchange.

[0094] In some embodiments, the anti-CD47 antibody may comprise HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region of SEQ ID NO: 1 and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region of SEQ ID NO: 2.

[0095] In some embodiments, the heavy chain variable region of the anti-CD47 antibody comprises or consists of an amino acid sequence of SEQ ID NO: 1 or an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence set forth in SEQ ID NO: 1.

[0096] In some embodiments, the light chain variable region of the anti-CD47 antibody comprises or consists of an amino acid sequence of SEQ ID NO: 2 or an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence set forth in SEQ ID NO: 2.

[0097] In some embodiments, the anti-CD47 antibody may comprise a heavy chain variable region of SEQ ID NO: 1 and / or a light chain variable region of SEQ ID NO: 2.

[0098] In some embodiments, the anti-CD47 antibody may comprise a heavy chain of SEQ ID NO: 3 and / or a light chain of SEQ ID NO: 4.

[0099] In some embodiments, the anti-CD47 antibody may be an antibody designated IMC-002. IMC-002 is a fully human IgG4 anti-CD47 antibody comprising a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 1 and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 2. IMC-002 comprises a S228P mutation as a hinge region mutation. The amino acid sequence information of IMC-002 is provided in Table 1 below.

[0100] 구분서열서열번호중쇄 가변 도메인QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSTLWVSEFDYWGQGTLVTVSS1경쇄 가변 도메인QSALTQPASVSGSPGQSISISCTGTSSDVGGHNYVSWYQQHPGKAPKLMIYDVRNRPSGVSNRFSGSKSGSTASLTISGLQTEDEADYYCNSYTGSRTYVFGSGTKLTVL2중쇄QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSTLWVSEFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK3경쇄QSALTQPASVSGSPGQSISISCTGTSSDVGGHNYVSWYQQHPGKAPKLMIYDVRNRPSGVSNRFSGSKSGSTASLTISGLQTEDEADYYCNSYTGSRTYVFGSGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS4중쇄 가변 도메인 CDR1GYTFTSY5중쇄 가변 도메인 CDR2NPSGGS6중쇄 가변 도메인 CDR3STLWVSEFDY7경쇄 가변Domain CDR1TGTSSDVGGHNYVS8Light chain variable domain CDR2DVRNRPS9Light chain variable domain CDR3NSYTGSRTYV10

[0101] IMC-002 is an antibody selected from among various anti-CD47 antibodies disclosed in Korean Patent Publication No. 10-2625835 (International Patent Application No. PCT / US2016 / 020980) that exhibited low hemagglutination in an in vitro hemagglutination assay, and is the C47B10-H3-D4 clone antibody disclosed in the aforementioned document. The entire contents of the aforementioned document are incorporated herein by reference.

[0102] In some embodiments, the dosage of the anti-CD47 antibody may vary depending on the age and weight of the subject being administered, the target disease, the condition, the route of administration, etc. When the anti-CD47 antibody is used to treat or inhibit the proliferation of solid tumors, the anti-CD47 antibody may be administered in single or multiple doses. The frequency and duration of treatment may be adjusted depending on the severity of the condition.

[0103] In some embodiments, the anti-CD47 antibody can be administered to the tumor of the subject one or more times, for example, one, two, three, four, five, six, seven, eight, nine, or ten or more times, in a therapeutically effective amount. For example, the therapeutic dosing regimen can include one or more administrations of the anti-CD47 antibody spaced about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks apart. In some embodiments, the anti-CD47 antibody can be administered about every 2 weeks to about 3 weeks. In some embodiments, the anti-CD47 antibody can be administered about every 2 weeks or about every 3 weeks.

[0104] In some embodiments, each dose of the anti-CD47 antibody can be about 5 to about 30 mg / kg. For example, each dose of the anti-CD47 antibody can be about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, or about 40 mg / kg. In some embodiments, each dose of the anti-CD47 antibody can be about 20 to about 30 mg / kg. In some embodiments, each dose of the anti-CD47 antibody is about 20 mg / kg or about 30 mg / kg.

[0105] In some embodiments, the lowest effective concentration of the anti-CD47 antibody for the treatment of a solid tumor may be about 20 to about 30 μg / mL. In some embodiments, the lowest effective concentration of the anti-CD47 antibody for the treatment of a solid tumor may be about 24 μg / mL. The lowest effective concentration is the minimum concentration of the drug that must be maintained in the serum for the treatment of a solid tumor. In some embodiments, administration of the pharmaceutical composition of the present invention may result in a serum concentration of the drug that is greater than or equal to the lowest effective concentration of about 20 to about 30 μg / mL. In some embodiments, administration of the pharmaceutical composition of the present invention may result in a serum concentration of the drug that is greater than or equal to the lowest effective concentration of about 24 μg / mL.

[0106] Pharmaceutical composition

[0107] In another aspect of the present invention, the anti-CD47 formulations disclosed herein can be provided as pharmaceutical compositions suitable for therapeutic use, e.g., human treatment. In some embodiments, the anti-CD47 antibodies disclosed herein can be formulated with one or more carriers and / or excipients to provide a pharmaceutical composition. In some embodiments, a pharmaceutical composition comprising an anti-CD47 antibody disclosed herein and one or more carriers and / or excipients is provided. Specifically, a therapeutic formulation comprising an anti-CD47 antibody can be prepared for storage by mixing an anti-CD47 antibody having a desired degree of purity with an optional physiologically acceptable carrier, excipient, or stabilizer in the form of a lyophilized formulation or aqueous solution. For a discussion of carriers, excipients, stabilizers, and the like, see Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980). The preferred form of the composition will depend on the intended route of administration and therapeutic application. In some embodiments, the composition may also include a pharmaceutically acceptable, non-toxic carrier or diluent, defined as a vehicle commonly used in formulating pharmaceutical compositions for animal or human administration, depending on the desired formulation. The diluent is selected so as not to affect the biological activity of the combination. Examples of diluents include distilled water, physiological phosphate-buffered saline, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also include other carriers, adjuvants, or non-toxic, non-therapeutic, non-immunogenic stabilizers, etc.

[0108] Typically, pharmaceutical compositions are prepared as injectables as liquid solutions or suspensions, although solid forms suitable for solution or suspension in liquid vehicles prior to injection may also be prepared. Furthermore, as discussed above, the formulations may be emulsified or encapsulated in liposomes or microparticles such as polylactides, polyglycolides, or copolymers for enhanced adjuvant effect. The formulations of the present invention may be administered in the form of depot injections or implant formulations, which may be formulated in a manner that allows for sustained or pulsatile release of the active ingredient. The pharmaceutical compositions are generally formulated aseptically, substantially isotonic, and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration.

[0109] In some embodiments, the pharmaceutical composition may be an aqueous solution composition.

[0110] In some embodiments, the pharmaceutical composition may comprise an anti-CD47 antibody of the invention, together with a phosphate buffer comprising about 10 to 30 mM sodium phosphate, about 80 to 120 mM sodium chloride, about 50 to 80 mM mannitol, and about 0.01 to 0.1% polysorbate 80.

[0111] In some embodiments, the pharmaceutical composition may comprise about 20 mM sodium phosphate, about 100 mM sodium chloride, about 65.9 mM mannitol, and about 0.05% polysorbate 80, together with an anti-CD47 antibody of the invention.

[0112] In some embodiments, the pH of the pharmaceutical composition may be about 4 to 7, specifically about 5 to 6, more specifically about 5.5.

[0113] In some embodiments, the pharmaceutical composition may comprise an anti-CD47 antibody of the invention at a concentration of about 40 to 60 mg / mL, specifically at a concentration of about 50 mg / mL.

[0114] In some embodiments, the anti-CD47 antibody of the present invention may be provided as a liquid formulation in a 10 mL sterile glass vial.

[0115] The anti-CD47 agent can be administered by any suitable route and method, including oral or parenteral administration. In some embodiments, the anti-CD47 agent is administered parenterally. Parenteral administration refers to any route of administration other than enteral and topical administration, typically by injection, and includes epidermal, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratendinous, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, intracranial, intrathoracic, epidural, and intrasternal injection and infusion. In some embodiments, the route of administration of the anti-CD47 agent is intravenous injection or infusion. In some embodiments, the route of administration of the anti-CD47 agent is intravenous infusion.

[0116] Combination administration

[0117] In one embodiment of the present invention, the anti-CD47 antibody of the present invention may be administered in combination with another anticancer agent. For example, the other anticancer agent may include, but is not limited to, one or more of the following: a chemotherapeutic agent (e.g., carboplatin, gemcitabine, paclitaxel, or a combination thereof), a tyrosine kinase inhibitor (e.g., lenvatinib, regorafenib, cabozantinib, or a combination thereof).

[0118] In some embodiments, the anti-CD47 antibody of the invention may be administered in combination with a tyrosine kinase inhibitor, preferably lenvatinib, regorafenib, cabozantinib, or a combination thereof, more preferably lenvatinib.

[0119] In some embodiments, the anti-CD47 antibody of the invention may be administered in combination with a chemotherapeutic agent, preferably paclitaxel, carboplatin, gemcitabine or a combination thereof, more preferably paclitaxel or carboplatin and gemcitabine.

[0120] In some embodiments, the anti-CD47 antibody may be administered in combination with lenvatinib to patients with hepatocellular carcinoma. In some embodiments, when administered in combination, lenvatinib may be administered once daily. In some embodiments, when administered in combination, lenvatinib may be administered in a dose of 4 to 8 mg (e.g., an 8 mg dose) once daily (body weight <60 kg) or in a dose of 4 to 12 mg (e.g., a 12 mg dose) once daily (body weight ≥ 60 kg).

[0121] In some embodiments, the anti-CD47 antibody may be administered in combination with paclitaxel to patients with triple-negative breast cancer. In some embodiments, when administered in combination, paclitaxel may be administered on days 1 to 3 of each cycle, preferably on day 1 of each cycle. In some embodiments, when administered in combination, paclitaxel is administered at a dose of 100 to 250 mg / m on days 1 to 3 of each cycle. 2 can be administered by IV infusion. For example, paclitaxel is administered at 175 mg / m on day 1 of each cycle. 2 It can be administered by IV.

[0122] In some embodiments, the anti-CD47 antibody may be administered in combination with gemcitabine and carboplatin to patients with triple-negative breast cancer. In some embodiments, when administered in combination, gemcitabine and carboplatin may each be administered 1 to 3 times, preferably 2 times, in each cycle. In some embodiments, when administered in combination, gemcitabine and carboplatin may each be administered on days 1 to 3 of each cycle and on days 6 to 10 of each cycle, respectively. In some embodiments, when administered in combination, gemcitabine and carboplatin may each be administered on days 1 and 8 of each cycle, respectively. In some embodiments, gemcitabine is administered at a dose of 800 to 1,200 mg / m on days 1 to 3 and on days 6 to 10 of each cycle. 2 Carboplatin can be administered IV at an AUC of 1.5 to 2.5. For example, gemcitabine can be administered at 1,000 mg / m on days 1 and 8 of each cycle. 2 It can be administered IV, and carboplatin can be administered IV at an AUC of 2.

[0123] Kit

[0124] In another aspect of the present method, a kit for using an anti-CD47 pharmaceutical composition disclosed herein is provided. The kit comprises an anti-CD47 agent disclosed herein. In some embodiments, the kit comprises two or more anti-CD47 agents. In some embodiments, the anti-CD47 agent is provided in a dosage form (e.g., a therapeutically effective dosage form). In some embodiments, the anti-CD47 agent is provided in two or more different dosage forms (e.g., two or more different therapeutically effective dosage forms). In some embodiments, the anti-CD47 agent can be provided in liquid or solid form in any convenient packaging (e.g., a stick pack, a dose pack, etc.).

[0125] In some embodiments, the kit may further include instructions for carrying out the method. The instructions for carrying out the method are the therapeutic or inhibitory methods disclosed herein. These instructions may be present in the kit in various forms, one or more of which may be present in the kit. One form in which these instructions may be present is information printed on a suitable medium or substrate, such as a piece or pieces of paper with the information printed on it, the packaging of the kit, a package insert, etc. Another form in which these instructions may be present is a computer-readable medium having the information recorded on it, such as a diskette, compact disc (CD), flash drive, etc. Another form in which these instructions may be present is a website address that can be used to access the information from a removed site via the Internet.

[0126] All patents and references cited herein are incorporated herein by reference in their entirety.

[0127] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended only to illustrate the present invention and the scope of the present invention is not limited to these examples.

[0128] Example

[0129] Example 1. Phase 1 dose-escalation and expansion study to evaluate the safety, tolerability, pharmacokinetics, and clinical activity of IMC-002 in patients with advanced and metastatic solid tumors.

[0130] This example describes a dose-escalation and expansion cohort study of the anti-CD47 antibody IMC-002 performed in patients with histologically or cytologically confirmed metastatic or locally advanced solid tumors who are refractory to standard therapy or who have progressed after standard therapy.

[0131] 1.1. Background and Basis

[0132] Pharmacological data from IMC-002 demonstrated its potential as an anticancer agent, and nonclinical safety, pharmacokinetic, and pharmacodynamic data were used to determine the starting dose for clinical trials. Furthermore, completed clinical trials in patients with metastatic or locally advanced solid tumors suggested that IMC-002 was safe and well-tolerated when administered weekly or biweekly.

[0133] CD47 is a transmembrane glycoprotein ubiquitously expressed on the surface of various cell types and overexpressed in various cancer cells. IMC-002 is a fully human IgG4 antibody targeting CD47 that lacks Fc modulator functions, including complement-dependent cellular cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC). IMC-002 binds to various cancer cells, including MDA-MB-231 (EC50 = 5.73 μg / mL) and Raji (EC50 = 4.77 μg / mL), but, unlike other CD47 blockers, it did not bind to red blood cells up to a concentration of 300 μg / mL and thus did not induce hemagglutination.

[0134] An in vitro phagocytosis assay was used to determine whether IMC-002 could induce antibody-dependent cellular phagocytosis (ADCP). As a single agent, IMC-002 induced phagocytosis in a dose-dependent manner against various cancer cell lines derived from solid tumors as well as hematological tumors.

[0135] The in vivo efficacy of IMC-002 was evaluated in a mouse xenograft model. In an efficacy study using the MDA-MB-231 xenograft model, administration of 3 mg / kg or more at weekly intervals significantly controlled cancer cell growth and prolonged survival. Furthermore, in vivo efficacy of IMC-002 was confirmed in the MDA-MB-231 orthotopic xenograft model at 10 mg / kg administered three times weekly. No drug-induced weight loss or abnormal clinical signs were observed in either test model. The pharmacokinetic (PK) characteristics of IMC-002 were evaluated in mice and monkeys. IMC-002 was administered intravenously to C57BL / 6 mice and cynomolgus monkeys at doses ranging from 0.4 to 10.0 mg / kg. IMC-002 exhibited overall dose-proportional systemic exposure. The steady-state volume of distribution (Vdss) was 75.1–83.8 mL / kg in mice and 137.9–200.3 mL / kg in monkeys. Total body clearance (CL) was 0.2–0.3 mL / hr / kg in mice and 0.7–1.1 mL / hr / kg in monkeys. Serum concentrations decreased with polyphasic pharmacokinetics, with a terminal elimination half-life (t1 / 2) of 224.9–240.9 hours in mice and 114.8–171.6 hours in monkeys.

[0136] In a 15-day preliminary toxicity study in monkeys, IMC-002 100 mg / kg administered intravenously three times at one-week intervals was well tolerated, and decreases in red blood cell and platelet counts were minimal. In a 4-week repeated-dose toxicity study in monkeys, IMC-002 10, 30, and 100 mg / kg were administered intravenously at one-week intervals for four weeks, followed by a four-week recovery period to evaluate reversibility. No abnormalities were found in clinical signs, general behavior and neurological function, body weight, food intake, eyes, electrocardiogram, respiratory rate, blood pressure, urinalysis, clinical chemistry, cytokine analysis, autopsy results, organ weights, or histopathology. Although drug-related changes were observed in hematology and peripheral blood immunophenotyping, they were of low toxicological significance, and individual values ​​were within or slightly outside the reference range. No related changes were observed in clinical or histopathological analyses. As a result, the no-observable adverse effect level (NOAEL) of IMC-002 was confirmed to be 100 mg / kg.

[0137] In vivo safety pharmacology studies evaluated the adverse effects of IMC-002 on the central nervous system, respiratory system, and cardiovascular system in a 4-week toxicity study in monkeys. No adverse drug-related changes in the nervous, respiratory, or cardiovascular systems were observed.

[0138] 1.2. IMC-002 formulation

[0139] The active pharmaceutical ingredient is IMC-002, a fully human anti-CD47 monoclonal antibody of the IgG4 kappa isotype. IMC-002 consists of a glycosylated tetramer composed of two identical 446-amino acid gamma heavy chains and two identical 216-amino acid kappa light chains linked by disulfide bonds. The IMC-002 drug product is provided as a liquid dosage form for IV infusion. IMC-002 is provided as a solution in a 10-mL sterile glass vial containing the antibody at a concentration of 500 mg / 10 mL (50 mg / mL) in a phosphate buffered saline solution (sodium phosphate 20 mM, sodium chloride 100 mM, mannitol 65.9 mM, 0.05% polysorbate 80, pH 5.5). IMC-002 was diluted with 5% Dextrose Injection and then injected into the subjects.

[0140] The drug substance (DS) of IMC-002 was manufactured by Samsung Biologics (Korea), and the drug product (DP) was manufactured by Vetter Pharma (US).

[0141] During the first four doses, all subjects were administered an antihistamine and paracetamol (e.g., 25-50 mg diphenhydramine and 500-650 mg paracetamol [acetaminophen] intravenously or an oral equivalent) approximately 30-60 minutes before each dose (or as per local standards), and the investigator decided whether to continue premedication in subjects who did not experience an infusion-related reaction thereafter. The medications administered were modified according to local treatment standards and guidelines.

[0142] 1.3. Clinical trial design

[0143] This clinical trial aimed to evaluate the safety, tolerability, pharmacokinetics, and clinical activity of IMC-002 in patients with advanced cancer who had failed standard treatment. It was conducted in two phases: a dose-escalation study (Part 1) and an expansion study (Part 2), as shown in Figure 2. In the expansion study, IMC-002 was administered in combination with other anticancer agents that patients had not previously received.

[0144] 1.3.1. Capacity-Step Escalation Study (Part 1)

[0145] A traditional 3+3 design was used to evaluate dose-limiting toxicities (DLTs) over 21 days, administering four doses of IMC-002 (5, 10, 20, and 30 mg / kg). IMC-002 was administered intravenously over a 3-hour (±30-minute) period (i.e., 150 to 210 minutes) every 2 weeks until disease progression or unacceptable toxicity. Tumor assessments were performed every 6 weeks according to RECIST 1.1 criteria.

[0146] Patients with metastatic or locally advanced solid tumors, histologically or cytologically confirmed, were selected for inclusion if they were ineligible for standard treatment or had not progressed to standard treatment. Furthermore, patients with at least one measurable lesion according to Response Evaluation Criteria in Solid Tumors version (RECIST) 1.1 and an ECOG performance status of 0–1 were included. RECIST is a well-known solid tumor response evaluation criterion as described by Eisenhauer et al. (European J. Cancer 45: 228-247 (2009). The Eastern Cooperative Oncology Group (ECOG) performance status index can be used to evaluate or select subjects for treatment, e.g., subjects with poor performance from prior therapy (see, e.g., Oken et al., (1982) Am J Clin Oncol. 5:649-655). The ECOG scale of performance status describes the patient's functional level in terms of ability to care for themselves, activities of daily living, and physical ability (e.g., walking, working, etc.).

[0147] Twelve refractory patients (9 with hepatocellular carcinoma, 2 with breast cancer, and 1 with gallbladder cancer) who met the above criteria were selected, with a mean age of 57 years (range, 48–73 years). Three patients were enrolled in each dose level 1–3 (5 mg / kg, 10 mg / kg, and 20 mg / kg), and no DLTs were observed. A total of four patients were enrolled in dose level 4 (30 mg / kg) because one patient was ineligible and not included in the DLT population. The analysis was performed in the DLT population. Eleven of these patients were stage IV patients with multiple distant metastases and a high tumor burden. The median number of prior systemic therapies was three (range, 1–3), and four patients had received anti-PD-(L)1 antibodies. The median interval between the last dose of prior therapy and the administration of IMC-002 was 1 month (range, 1–4). The baseline characteristics of the selected patients are summarized in Table 2 below.

[0148] Tendency 5 mg / kg 10 mg / kg 20 mg / kg 30 mg / kg Total (n=3) (n=3) (n=3) (n=3) (N=12) Age, Year * 56 (54-57) 62 (53-73) 53 (52-73) 60 (48-64) 57 (48-73) Sex, n Male 31228 Female 02114 Race, Asian 333312 ECOG PS 0 233311 Primary Disease 32229 Breast Cancer 101012 Gallbladder Cancer 00101 Disease Stage IV 332311 Surgical History 332311 RT History 31138# Number of Previous Systemic Treatments 10 200 220 111 3330 227 Previous Anti-PD-(L)1 227 n20114 Interval between IMC-002 doses in previous treatment, months*1.0 (0.7-1.0)1.0 (1.0-1.6)1.0 (1.0-3.9)1.5 (1.3-1.5)1.0 (0.7-3.9)

[0149] 1.3.2 Extension Study: Combination Administration (Part 2)

[0150] Nine patients with hepatocellular carcinoma (HCC) and one patient with triple-negative breast cancer (TNBC) were enrolled as trial subjects, and eight patients had prior anti-PD-(L)1 therapy. In the extension study, IMC-002 was administered in combination with previously untreated chemotherapy agents. Lenvatinib was used in the combination therapy for the HCC cohort, and gemcitabine plus carboplatin was used in the combination therapy for the triple-negative breast cancer cohort.

[0151] All subjects received IMC-002 20 mg / kg IV every 3 weeks (Q3W). In the first cycle, IMC-002 was administered over 3 hours (±30 minutes). If the first infusion was tolerated, subsequent infusions were administered over 1 to 1.5 hours (±10 minutes). Standard of care (SOC, lenvatinib, paclitaxel, or gemcitabine plus carboplatin) was initiated after completion of the 1-hour safety monitoring for IMC 002. IMC-002 was administered for up to 2 years (34 cycles) until progressive disease (PD) or unacceptable toxicity occurred.

[0152] In the hepatocellular carcinoma cohort, lenvatinib was administered once daily at the clinically approved dose of 8 mg (two 4-mg capsules) in subjects weighing less than 60 kg or 12 mg (three 4-mg capsules) in subjects weighing 60 kg or more. Lenvatinib was administered for up to 2 years (34 cycles) until progressive disease or unacceptable toxicity.

[0153] In the triple-negative breast cancer cohort, subjects received gemcitabine plus carboplatin for up to 2 years (34 cycles) until progressive disease (PD) or unacceptable toxicity. Gemcitabine 1,000 mg / m on days 1 and 8 of each cycle. 2After administration of gemcitabine, carboplatin AUC 2 was administered intravenously. The dosage of carboplatin was applied according to the following formula. Gemcitabine was administered over approximately 30 minutes, and carboplatin was administered over approximately 30 to 60 minutes.

[0154] Carboplatin dose (mg) = AUC × (GFR + 25)

[0155] GFR: Glomerular filtration rate (not converted to body surface area)

[0156] The baseline characteristics of the selected patients are summarized in Table 3 below.

[0157] Total (N=10) Age, year *61 Sex, n Male 9 Female 1 Race, Asian 10 ECOG PS 0, n Primary disease, n Hepatocellular carcinoma 9 Triple-negative breast cancer 1 Disease stage IV, n9 History of surgery, n4 History of RT, n6# Number of previous systemic treatments ≥ 24 Previous anti-PD-(L)1, n8

[0158] 1.4. Safety Results

[0159] 1.4.1. Capacity-escalation study (Part 1)

[0160] No dose-limiting toxicities (DLTs) were observed across all four dose levels. The majority of treatment-related adverse events (TRAEs) were grade 1-2 (92%), and most of these events occurred in the first cycle (95%). TRAEs observed in at least two patients included transient scleroderma, skin rash, anemia, and nausea. No infusion-related reactions, thrombocytopenia, or neutropenia were reported. The rates of treatment-related adverse events in the safety population are summarized in Table 4.

[0161] Adverse Effect Terminology 5 mg / kg (n=3) 10 mg / kg (n=3) 20 mg / kg (n=3) 30 mg / kg (n=3) Total (N=12) Anemia 235 (42%) Decreased appetite 11 (8%) Diarrhea 11 (8%) Fatigue 11 (8%) Headache 112 (17%) Increased liver function test values ​​11 (8%) Muscle pain 22 (17%) Nausea 112 (17%) Non-cardiac chest pain 11 (8%) Pruritus 11 (8%) Fever 11 (8%) Skin rash 23229 (75%) Rash 3328 (67%) Vomiting 11 (8%)

[0162] Hemoglobin, neutrophil, and platelet counts were measured after IMC-002 administration. Although hemoglobin levels decreased slightly after initial exposure to IMC-002, they subsequently recovered without evidence of hemolytic anemia. Recovery was delayed in the 30 mg / kg cohort compared to the 20 mg / kg cohort. Based on this, the 20 mg / kg dose was administered in the expansion study (Part 2). Neutropenia and thrombocytopenia, specific side effects of anti-CD47 agents, were not observed after initial exposure. Figures 3 to 5 demonstrate that hemoglobin, platelet, and neutrophil counts remained above the lower limit of normal (LLN) after IMC-002 administration.

[0163] 1.4.2. Extended Study (Part 2)

[0164] A total of 10 patients were treated with a 3-week regimen and combination therapy. Anemia was reported in one patient as a specific side effect of the anti-CD47 agent. Skin rash and erythema were observed at a lower rate than in the dose-escalation study (Part 1). The rates of treatment-related adverse reactions in the safety population are summarized in Table 5.

[0165] Adverse reaction terms 20 mg / kg (N=10) Anemia 1 (10%) Diarrhea 1 (10%) Fatigue 1 (10%) Fever 1 (10%) Asthenia 3 (30%) Headache 1 (10%) Indigestion 1 (10%) Infusion-related reaction 1 (10%) Skin rash 4 (40%) Tinnitus 1 (10%) Thyroid-stimulating hormone increase 1 (10%)

[0166] Hemoglobin, neutrophil, and platelet counts were measured after IMC-002 administration. No grade 2 anemia was observed during the 3-week treatment period. Anemia, neutropenia, and thrombocytopenia, which are specific side effects of anti-CD47 agents, were also not observed after initial exposure. Figures 12 to 14 demonstrate that hemoglobin, platelet, and neutrophil counts remained above the lower limit of normal (LLN) after IMC-002 administration.

[0167] 1.5. Validity Results

[0168] 1.5.1. Capacity-escalation study (Part 1)

[0169] Serum exposure (Cmax and AUC) of IMC-002 increased in a dose-dependent manner, and the predicted trough concentration exceeded the lowest efficacy concentration of IMC-002 when administered at doses ≥10 mg / kg once every 2 weeks. Of the 12 patients eligible for efficacy evaluation, 6 had stable disease (Disease Control Rate 50%) after a median treatment period of 11 cycles (range 6–28), and 4 maintained stable disease for ≥6 months (Clinical Benefit Rate 30%). Five of these patients had hepatocellular carcinoma and 1 had breast cancer. See Figure 6 for a swimmer plot of tumor response over time. The change in tumor burden from baseline to nadir for target lesions is shown in Figure 7. Referring to the waterfall plot in Figure 7, it can be seen that 3 of the 12 patients experienced a decrease in tumor size (up to -20%).

[0170] Among the 12 patients eligible for efficacy evaluation, the disease control rate was 50.0%, the clinical benefit rate (CBR; stable disease for ≥6 months of treatment) was 33.3%, and the median treatment duration was 10 months. CD47 immunohistochemistry (IHC) images were analyzed using an AI-based analysis tool (Lunit SCOPE) that can distinguish staining positivity and cell types at the single-cell level. Specifically, the AI-based analysis tool used the Lunit SCOPE uIHC v.2 model.

[0171] In the AI ​​analysis of CD47 IHC, the density of CD47-positive macrophages tended to be higher in cases with clinical benefit than in cases without (mean CD47+ macrophage density: 71.0 vs 44.3 cells / mm 2 ), the proportion of tumor cells expressing CD47 was similar between the two groups (mean CD47+ tumor cells: 3289.3 vs 4098.4 cells / mm 2 ). Figures 8A and 8B show the CD47-positive macrophage density and CD47-negative macrophage density in patients who achieved clinical benefit and those who did not. The group of patients who achieved clinical benefit showed a higher density of CD47-positive macrophages. Table 6 below shows the analysis results of the four patients who showed clinical benefit.

[0172] Subject (gender / age) Cohort (mg / kg) Diagnosis Change in target lesion % Period (months) CD47 IHC (%) CD47+ cancer cell density (cells / mm 2 )CD47-cancer cell density (cells / mm 2 )CD47+ macrophage density (cells / mm 2 )CD47-macrophage density (cells / mm 2)A(M / 56)5HCC-17.246615876.33784.09.20.4I(M / 73)20HCC9.5211395.03339.917.98.1J(M / 48)30HCC-14.638843344.3624.0196.59.1K(M / 64)30HCC-20.0018703841.71627.760.41.2

[0173] 1.5.2. Capacity Determination through PK Modeling

[0174] Population pharmacokinetic (PK) analyses were performed using NONMEM (version 7.5, ICON Development Solutions, Ellicott City, MD, USA) with support from PsN (version 3.0.0, Perl-speaks-NONMEM, Uppsala, Sweden). A total of 213 unbound IMC-002 serum concentration samples from patients were analyzed for model development. The first-order conditional estimation with interaction (FOCE-I) method was used for parameter estimation, and model selection was based on numeric (e.g., statistically significant decrease in objective function value (OFV)) and visual (e.g., goodness-of-fit (GOF) plots and visual predictive testing (VPC, n=1000)) criteria. Bootstrapping (n=1000) was performed for nonparametric model evaluation. Structural models explored general compartment models (e.g., 1-, 2-, and 3-compartment models) and target-mediated drug dispensing (TMDD) models via neonatal Fc receptor (FcRn) recycling. Stepwise covariate modeling (SCM) was used to identify clinically relevant covariates. Simulation studies of the final model were performed under various scenarios to optimize dose for clinical trials.

[0175] After considering numerical and visual criteria, the TMDD model including FcRn recycling of IgG was selected as the final model because it adequately described the pharmacokinetics of IMC-002. In addition, physiological model parameters (e.g., CD47 degradation rate, total FcRn receptor abundance) and target binding affinity (e.g., CD47 binding affinity, FcRn binding affinity, dissociation rate of drug-CD47 complex) were fixed in the final model based on literature and in vitro observations. There was no covariate effect on the PK of IMC-002. Simulations were performed using the final model for the optimal dose considering the lowest effective concentration (MEC, 24 μg / mL) obtained in the nonclinical efficacy study. Consequently, the 3-weekly dosing regimen (Q3W) was considered an equivalent efficacy regimen compared to the 2-weekly dosing regimen (Q2W) performed in the previous clinical trial. Key parameters at steady state for evaluating the equivalent efficacy regimen are presented in Table 7 and Figure 9. AUC in Table 7 tau means the area under the serum concentration curve at the dosing interval, MEC means the lowest effective concentration, and C trough refers to the minimum serum concentration of the drug. Figure 9 shows the simulated pharmacokinetic (PK) profile of IMC-002 after IV infusion every 3 weeks (Q3W), where the solid line represents the median PK profile, the shaded area represents the 5th to 95th percentile PK profiles, and the dashed line represents the lowest effective concentration (MEC, 24 μg / mL). Figure 10 shows the comparative results of PK profiles after intravenous administration every 3 weeks according to the administered dose. Figure 11 shows the PK serum concentration after the initial administration. Table 7 shows the PK simulation results for the 3-week regimen in the 5th cycle (cycle 5) when steady state was confirmed.

[0176] Parameter 5 mg / kg 10 mg / kg 20 mg / kg 30 mg / kg AUC tau(μgh / ml)9381 ± 312324035 ± 1182861847 ± 32098103366 ± 46690AUC tau / MEC (μgh / ml)2349 ± 171312993 ± 1101049465 ± 3214790984 ± 46827C trough (μg / mL)6.1 ± 2.317.1 ± 11.855.9 ± 46.2109.2 ± 74.3C trough over MEC (%)-148099

[0177] IMC-002 is an engineered fully human IgG4 monoclonal antibody. Because IgG exhibits a longer half-life than other immunoglobulins due to recycling of the neonatal Fc receptor (FcRn) in the hemolymph, IMC-002 exhibited characteristics similar to endogenous IgG. This phenomenon is attributed to the nonlinear PK of the drug, which is why a mechanism-based TMDD model was selected as the final model. Because IMC-002 was administered at a weight-based dose and all tested covariates (e.g., weight, age, and sex) could be correlated with weight, no statistically significant covariates were expected. Based on the simulation results, considering the plasma concentration retention time and maximum tolerated dose (MTD) for MEC observed in nonclinical and clinical studies, a 20 mg / kg Q3W regimen was suggested as an effective and safe equivalent dosing regimen. Additionally, the half-life of IMC-002 is estimated to be approximately 4 to 5 days, which provides the advantage of synchronizing the dosing frequency with other anticancer agents in a clinical setting. Since chemotherapy cycles typically last three weeks, a half-life of 4 to 5 days is expected to maintain consistent systemic exposure without accumulation.

[0178] 1.5.3. Extended Study (Part 2)

[0179] As a result of administering IMC-002 at a dose of 20 mg / kg once every 3 weeks (combination administration), among 9 patients with liver cancer, 1 showed a partial response and 5 showed stable disease, resulting in a disease control rate of 67%. Of these, 3 patients maintained a partial response or stable disease for more than 6 months, resulting in a clinical benefit rate of 33%. Refer to Figure 15 for a swimmer plot of tumor response over time. Figure 16 shows the change in tumor burden from baseline to the nadir for the target lesion. Referring to the waterfall plot of Figure 16, it can be seen that a decrease in tumor size (up to -31%) was observed in 6 out of 10 patients. In the case of a patient with triple-negative breast cancer (Figure 16 (A)), it can be seen that the tumor size was reduced by 12%.

[0180] conclusion

[0181] Dose-escalation studies demonstrated that IMC-002 had an excellent safety profile at doses up to 30 mg / kg every two weeks for up to 18 months and showed preliminary efficacy in patients with advanced or metastatic solid tumors whose disease had progressed following standard therapy.

[0182] Based on PK modeling and safety data, 20 mg / kg every 3 weeks was determined to be the optimal dose, and this dosing regimen was thought to allow for easier combination dosing.

[0183] We confirmed that IMC-002 monotherapy provided significant clinical benefit in advanced or metastatic solid tumors, particularly refractory HCC. Furthermore, we observed a higher density of CD47-positive macrophages in patients with CBR compared to non-CBR patients.

[0184] An extension study confirmed that IMC-002 exhibited enhanced safety and clinical activity when administered at a dose of 20 mg / kg every 3 weeks. IMC-002 demonstrated an excellent safety profile when administered at a dose of 20 mg / kg every 3 weeks, and clinical benefit was enhanced when combined with other drugs, such as lenvatinib, paclitaxel, or gemcitabine plus carboplatin.

Claims

1. A pharmaceutical composition for treating a tumor or inhibiting tumor proliferation in a patient with advanced or metastatic solid tumor, comprising an effective amount of a fully human anti-CD47 antibody that specifically binds to CD47 and comprises HCDR1, HCDR2 and HCDR3 contained in a heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in a light chain variable region (LCVR) of SEQ ID NO: 2, The antibody is administered to the patient at a dose of about 5 mg / kg to about 30 mg / kg every two to three weeks without priming. Pharmaceutical composition.

2. In paragraph 1, A pharmaceutical composition, wherein the heavy chain variable region comprises HCDR1 of SEQ ID NO: 5, HCDR2 of SEQ ID NO: 6, and HCDR3 of SEQ ID NO: 7, and the light chain variable region comprises LCDR1 of SEQ ID NO: 8, LCDR2 of SEQ ID NO: 9, and LCDR3 of SEQ ID NO:

10.

3. In paragraph 1, A pharmaceutical composition wherein the heavy chain variable region comprises an amino acid sequence of SEQ ID NO: 1 or an amino acid sequence at least 95% identical thereto, and the light chain variable region comprises an amino acid sequence of SEQ ID NO: 2 or an amino acid sequence at least 95% identical thereto.

4. In paragraph 1, The above antibodies do not cause significant levels of red blood cell aggregation or anemia in patients. Pharmaceutical composition.

5. In paragraph 1, A pharmaceutical composition comprising about 10 to 30 mM sodium phosphate, about 80 to 120 mM sodium chloride, about 50 to 80 mM mannitol, and about 0.01 to 0.1% polysorbate 80, and having a pH of about 4 to 7.

6. In paragraph 1, A pharmaceutical composition comprising about 20 mM sodium phosphate, about 100 mM sodium chloride, about 65.9 mM mannitol and 0.05% polysorbate 80 and having a pH of about 5.

5.

7. In paragraph 1, A pharmaceutical composition, wherein the composition is in a liquid dosage form for intravenous injection.

8. In paragraph 1, A pharmaceutical composition wherein the solid cancer is liver cancer, breast cancer or gallbladder cancer.

9. In paragraph 1, A pharmaceutical composition wherein the above solid cancer is hepatocellular carcinoma.

10. In paragraph 1, A pharmaceutical composition wherein the above solid tumor is triple negative breast cancer.

11. In paragraph 1, A pharmaceutical composition for a patient who is unable to receive standard treatment or has failed standard treatment.

12. In paragraph 1, A pharmaceutical composition wherein the antibody is administered to the patient about every two or three weeks.

13. In paragraph 1, A pharmaceutical composition wherein the antibody is administered to the patient at a dose of about 20 mg / kg or about 30 mg / kg.

14. In paragraph 1, The above patient had received at least one systemic treatment prior to administration of anti-CD47 antibody. Pharmaceutical composition.

15. In paragraph 1, The above patient is a stage 4 patient with multiple distant metastases or a locally advanced patient. Pharmaceutical composition.

16. In paragraph 1, The above antibody is administered in combination with other anticancer drugs. Pharmaceutical composition.

17. In paragraph 16, The other anticancer drugs mentioned above are lenvatinib, paclitaxel, gemcitabine, carboplatin or a combination thereof. Pharmaceutical composition.

18. In paragraph 17, The above lenvatinib is administered once a day. Pharmaceutical composition.

19. In paragraph 17, The above paclitaxel is administered on the first day of the administration cycle of the above antibody. Pharmaceutical composition.

20. In paragraph 17, The above gemcitabine and carboplatin are administered on the 1st and 8th days of the administration cycle of the above antibody. Pharmaceutical composition.

21. As a method for treating tumors or inhibiting tumor growth, (a) selecting a patient with advanced or metastatic solid tumor; and (b) administering an anti-CD47 antibody to the patient at a dose of about 5 mg / kg to about 30 mg / kg without priming every two to three weeks. The above antibody is an antibody that specifically binds to CD47 and includes HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region of SEQ ID NO:

2. method.

22. In paragraph 21, further comprising a step of measuring CD47 positive macrophage density in a biological sample obtained from a patient before or after administration of the anti-CD47 antibody. method.

23. In paragraph 21, (c) further comprising a step of additionally administering another anticancer agent before or after administration of the antibody, or simultaneously with administration of the antibody. method.

24. In paragraph 23, The above other anticancer agent is administered once a day, or is administered on the first day of the administration cycle of the antibody, or is administered on the first and eighth days of the administration cycle of the antibody. method.

25. A kit comprising an anti-CD47 antibody together with instructions for use of the anti-CD47 antibody for treating a tumor or inhibiting tumor growth in a patient with a solid tumor, The instructions for use include instructions for administering the antibody to the patient at a dose of about 5 mg / kg to about 30 mg / kg every two to three weeks without priming, The above antibody is an antibody that specifically binds to CD47 and includes HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region of SEQ ID NO:

2. Kit.

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