Pharmaceutical composition for prevention or treatment of cancer, comprising BTK degrader and mettl3 inhibitor

Abstract

The present invention relates to a pharmaceutical composition for prevention or treatment of cancer, comprising a BTK degrader and a METTL3 inhibitor, for which it has been confirmed that when a BTK degrader and a METTL3 inhibitor based on proteolysis-targeting chimera (PROTAC) technology are co-administered, a synergistic effect of exhibiting notable anticancer activity is achieved compared to when same are used alone, and thus a prevention or treatment effect on various cancers such as leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer may be created.

Description

Pharmaceutical composition for preventing or treating cancer comprising a BTK decomposer and a METTL3 inhibitor

[0001] This patent application claims priority to Republic of Korea Patent Application No. 10-2023-0177297, filed with the Korean Intellectual Property Office on December 8, 2023, the disclosure of which is incorporated herein by reference.

[0002] The present invention was made possible by the support of the Ministry of Science and ICT under the project identification number 1711172425, project number 2022R1F1A1061005 (Industry-Academia Cooperation Group Number: 202203890001). The research management specialized organization of the project is the Industry-Academia Cooperation Group of Inje University, the research project name is "Basic Research", the research project name is "Development of a combination immunotherapy using a PROTAC-based epigenetic regulator EZH2 target drug and an immune checkpoint inhibitor for the treatment of refractory lymphoma", the main organization is the Industry-Academia Cooperation Group of Inje University, and the research period is 2022.06.01 ~ 2025.02.28.

[0003] The present invention relates to a pharmaceutical composition for preventing or treating cancer comprising a BTK degrader and a METTL3 inhibitor, and more particularly, to a combination therapy use of a BTK degrader and a METTL3 inhibitor for preventing or treating cancer.

[0004] Leukemia is a group of life-threatening malignant diseases of the blood and bone marrow. Genetic errors such as reciprocal chromosomal translocations, chromosomal deletions, point mutations, and epigenetic changes can inhibit stem cell maturation throughout various stages of hematopoiesis, leading to the excessive proliferation of immature white blood cells. Leukemias are classified as myeloid and lymphoid, depending on their cell origin. Leukemia is a cancer that causes a clonal expansion of white blood cells in the bone marrow, increases the number of affected lineage cells in the circulation, and is also a lymphoid malignancy, a type of cancer that causes abnormal cell proliferation in lymphoid tissue, resulting in the suppression of normal white blood cell, red blood cell, and platelet production. A decrease in white blood cell count can lead to immune suppression, leading to sepsis caused by bacterial infection. A decrease in red blood cells can lead to anemia, and a decrease in platelets can lead to a bleeding tendency. Furthermore, hyperproliferating leukemic cells can cause fever, fatigue, pain, decreased consciousness, and a tendency to bleed. Leukemia is classified as acute or chronic depending on the degree of cell differentiation, i.e. the rate of deterioration.

[0005] Lymphoma is a blood cancer that occurs when B or T lymphocytes become cancerous. Lymphoma is a malignant tumor that develops in lymphoid tissue, but it can also develop in non-lymphoid tissue. Lymphoma is broadly classified as Hodgkin lymphoma and non-Hodgkin lymphoma, with Hodgkin lymphoma accounting for approximately 10% of all lymphomas, and non-Hodgkin lymphoma accounting for the remaining 90%. The most well-established risk factor for non-Hodgkin lymphoma is immunosuppression, with patients with HIV at increased risk for high-grade non-Hodgkin lymphoma. Other high-risk groups include organ transplant recipients, patients receiving high-dose chemotherapy in conjunction with stem cell transplantation, and those with inherited immunodeficiency syndromes or autoimmune diseases. Infections also play a significant role in the development of some lymphomas. Epstein-Barr Virus (EBV), a type of herpes virus, infects most people and causes an asymptomatic latent infection, but it is known to cause tumors such as Burkitt's lymphoma, nasopharyngeal carcinoma (NPC), and Hodgkin's disease in individuals with weakened immune systems. Helicobacter pylori infection has been identified as a risk factor for gastric mucosa-associated lymphoid tissue lymphoma. Existing treatments for lymphoma include reducing the dose of immunosuppressants, using antiviral agents, chemotherapy, and administration of rituximab antibodies. However, there are differences in treatment response rates, and an effective treatment method has not yet been established.

[0006] For the past several decades, conventional small molecule inhibitors (SMIs) with high cell penetration have been the primary therapeutic approach for cancer. These drugs exhibit efficacy by binding to the active site of a target protein and inhibiting its function. However, many proteins lack active sites or possess sites that are unsuitable for inhibitory action. Consequently, high doses are required to maximize therapeutic efficacy, often leading to drug resistance. To overcome these limitations, a strategy that has recently gained attention in clinical practice is PROteolysis Targeting Chimera (PROTAC) technology.

[0007] PROTAC technology, a targeted therapy for cancer cells, utilizes the ubiquitin-proteasome system (UPS) to selectively degrade target proteins. Research is currently underway on this technology. PROTAC induces ubiquitination of a target protein by positioning it in proximity to an E3 ubiquitin ligase, thereby promoting its natural degradation by the patient's own protein degradation system. This technology holds great potential as a treatment for various diseases, including blood cancer.

[0008] However, there are reports that PROTAC-based tumor protein degradation does not result in an effective level of tumor growth inhibition, or that the target protein degradation effect of PROTAC varies greatly depending on the tissue of administration. Therefore, there is a need to develop a PROTAC-based anticancer therapy that can exhibit an anticancer effect greater than that of conventional chemical drugs.

[0009] The B-cell receptor (BCR) plays a crucial role in regulating the proliferation and survival of various lymphomas, including chronic lymphocytic leukemia (CLL), non-Hodgkin lymphoma (NHL), mantle cell lymphoma (MCL), and diffuse large B-cell lymphoma (DLBCL). Bruton's tyrosine kinase (BTK), a member of the tyrosine kinase subfamily, is a key mediator of B-cell development and malignancy due to its ability to mediate signaling complexes at the B-cell receptor. Excessive activation of B-cell receptor (BCR) signaling promotes the proliferation, differentiation, and survival of malignant B cells, and disruption of BTK by specific inhibitors can interfere with downstream B-cell receptor signaling and thereby induce apoptosis of B-cell tumor cells. Therefore, small molecule inhibitors targeting BTK may be beneficial in the treatment of B-cell malignancies and autoimmune diseases.

[0010] METTL3 (N6-adenosine-methyltransferase complex catalytic subunit) regulates mRNA by transferring methyl groups during m6A modification, thereby promoting tumor initiation and progression. METTL3 inhibitors are expected to be effective in tumor-targeted therapy, as they inhibit this process and restore chemosensitivity of tumor cells in vitro.

[0011] Accordingly, the present inventors have confirmed that when a BTK degrader and a METTL3 inhibitor are administered in combination based on proteolysis-targeting chimera (PROTAC) technology, there is a synergistic effect that shows significant anticancer activity compared to when they are administered alone, and thus, it can create a preventive or therapeutic effect on various cancers such as leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer.

[0012] Accordingly, the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which comprises a BTK decomposer and a METTL3 inhibitor as active ingredients.

[0013] Another object of the present invention is to provide a combination kit for preventing or treating cancer comprising (i) a BTK degrader and (ii) a METTL3 inhibitor.

[0014] Another object of the present invention is to provide a method for preventing or treating cancer using a pharmaceutical composition comprising a BTK decomposer and a METTL3 inhibitor as active ingredients.

[0015] Another object of the present invention is to provide a use of a BTK degrader and a METTL3 inhibitor for the prevention or treatment of cancer.

[0016] Another object of the present invention is to provide a combination therapy use of a BTK degrader and a METTL3 inhibitor for the prevention or treatment of cancer.

[0017] The present invention relates to a pharmaceutical composition for preventing or treating cancer comprising a BTK decomposer and a METTL3 inhibitor, and leukemia, lymphoma, lung cancer, pancreatic cancer, or breast cancer can be effectively prevented or treated through the pharmaceutical composition of the present invention.

[0018] Hereinafter, the present invention will be described in more detail.

[0019] One aspect of the present invention relates to a pharmaceutical composition for preventing or treating cancer, comprising a BTK decomposer and a METTL3 inhibitor as active ingredients.

[0020] The term "BTK" in this specification refers to a member of the tyrosine kinase subfamily, a mediator that plays a crucial role in the development and malignancy of B cells due to its ability to mediate signaling complexes at the B-cell receptor. Excessive activation of B-cell receptor (BCR) signaling promotes the proliferation, differentiation, and survival of malignant B cells, and it has been shown that disruption of BTK by specific inhibitors can interfere with downstream B-cell receptor signaling and thereby induce apoptosis of B-cell tumor cells. Therefore, small molecule inhibitors targeting BTK are known to be beneficial in the treatment of B-cell malignancies and autoimmune diseases.

[0021] The term “BTK degrader (dBTK)” in this specification refers to an agent that targets and degrades the BTK protein.

[0022] The term "METTL3" in this specification refers to N 6 -methyladenosine (m 6 A) METTL3 refers to an RNA methyltransferase involved in the biosynthesis, degradation, and translation control of mRNA through modification. METTL3 promotes the translation of important oncogenes such as epidermal growth factor receptor (EGFR) and the Hippo pathway effector TAZ in human cancer cells, and is known to be required for cancer cell growth, survival, and invasion. Inhibiting METTL3 is expected to be effective in tumor-targeted therapy, as it restores chemosensitivity of tumor cells in vitro.

[0023] The term “METTL3 inhibitor (iMETTL3)” in this specification refers to an agent that inhibits the activity of METTL3.

[0024] The term "comprising as an active ingredient" in this specification means including an amount sufficient to achieve a specific effect of a BTK degrader or METTL3 inhibitor, for example, activity for preventing or treating cancer for the purposes of the present invention.

[0025] The term “prevention” in this specification means any action that inhibits or delays the progression of cancer by administering the pharmaceutical composition of the present invention.

[0026] The term “treatment” as used herein means inhibition of cancer development, alleviation of cancer, and elimination of cancer.

[0027] In the present invention, the composition may be for combined administration of a BTK decomposer and a METTL3 inhibitor.

[0028] In the present invention, the BTK decomposer may be based on PROTAC (Proteolysis targeting chimera).

[0029] In the present invention, the PROTAC molecule is a heterobifunctional compound composed of a ubiquitin ligase binding ligand, a target protein binding ligand, and a linker connecting them, which serves to bind an E3 ligase and a target protein, and then degrades the protein using a ubiquitin-based protein degradation system within the cell.

[0030] In one embodiment of the present invention, the BTK degrader is a PROTAC molecule.

[0031] In one embodiment of the present invention, the PROTAC-based BTK degrader may mean a substance aimed at degrading the BTK protein.

[0032] In the present invention, the BTK decomposer may be MT-802, but is not necessarily limited thereto.

[0033] In one embodiment of the present invention, the MT-802 is a compound having a structure represented by the following structural formula 1.

[0034] [Structural formula 1]

[0035]

[0036] In the present invention, the METTL3 inhibitor may be STM2457, but is not necessarily limited thereto.

[0037] In one embodiment of the present invention, the STM2457 is a compound having a structure of structural formula 2 below.

[0038] [Structural formula 2]

[0039]

[0040] The present inventors confirmed that when a BTK decomposition agent and a METTL3 inhibitor were used alone, the minimum molar concentration at which the drug was deemed effective was 5 μM for both drugs. Accordingly, when a BTK decomposition agent and a METTL3 inhibitor were used in combination, these results were reflected. When each drug was included in a pharmaceutical composition at a molar ratio of 1:1, the minimum molar concentration at which the drug was effective was 5 μM. As a result, compared to the single treatment, the combination treatment had a superior cancer cell growth inhibition effect, i.e., a superior anticancer activity, confirming that the combination treatment had a synergistic effect.

[0041] In one embodiment of the present invention, the molar ratio of the BTK degrader and the METTL3 inhibitor is 1:10 to 10:1, for example, 1:10 to 9:1, 1:10 to 8:1, 1:10 to 7:1, 1:10 to 6:1, 1:10 to 5:1, 1:10 to 4:1, 1:10 to 3:1, 1:10 to 2:1, 1:10 to 1:1, 1:9 to 10:1, 1:9 to 9:1, 1:9 to 8:1, 1:9 to 7:1, 1:9 to 6:1, 1:9 to 5:1, 1:9 to 4:1, 1:9 to 3:1, 1:9 to 2:1, 1:9 to 1:1, 1:8 to 10:1, 1:8 to 9:1, 1:8 to 8:1, 1:8 to 7:1, 1:8 to 6:1, 1:8 to 5:1, 1:8 to 4:1, 1:8 to 3:1, 1:8 to 2:1, 1:8 to 1:1, 1:7 to 10:1, 1:7 to 9:1, 1:7 to 8:1, 1:7 to 7:1, 1:7 to 6:1, 1:7 to 5:1, 1:7 to 4:1, 1:7 to 3:1, 1:7 to 2:1, 1:7 to 1:1, 1:6 to 10:1, 1:6 to 9:1, 1:6 to 8:1, 1:6 to 7:1, 1:6 to 6:1, 1:6 to 5:1, 1:6 to 4:1, 1:6 to 3:1, 1:6 to 2:1, 1:6 to 1:1, 1:5 to 10:1, 1:5 to 9:1, 1:5 to 8:1, 1:5 to 7:1, 1:5 to 6:1, 1:5 to 5:1, 1:5 to 4:1, 1:5 to 3:1, 1:5 to 2:1, 1:5 to 1:1, 1:4 to 10:1, 1:4 to 9:1, 1:4 to 8:1, 1:4 to 7:1, 1:4 to 6:1, 1:4 to 5:1, 1:4 to 4:1, 1:4 to 3:1, 1:4 to 2:1, 1:4 to 1:1, 1:3 to 10:1, 1:3 to 9:1, 1:3 to 8:1, 1:3 to 7:1, 1:3 to 6:1, 1:3 to 5:1, 1:3 to 4:1,1:3 to 3:1, 1:3 to 2:1, 1:3 to 1:1, 1:2 to 10:1, 1:2 to 9:1, 1:2 to 8:1, 1:2 to 7:1, 1:2 to 6:1, 1:2 to 5:1, 1:2 to 4:1, 1:2 to 3:1, 1:2 to 2:1, 1:2 to 1:1, 1:1 to 10:1, 1:1 to 9:1, 1:1 to 8:1, 1:1 to 7:1, 1:1 to 6:1, 1:1 to 5:1, 1:1 to 4:1, 1:1 to 3:1, or 1:1 to 2:1, for example, 1:3 to 3:1, but It is not limited to,

[0042] In one embodiment of the present invention, the pharmaceutical composition may comprise a BTK degrader at a concentration of 1 to 20 μM and a METTL3 inhibitor at a concentration of 1 to 20 μM.

[0043] In one embodiment of the present invention, the pharmaceutical composition may comprise a BTK degrading agent at a concentration of 1 to 20 μM, 1 to 15 μM, 1 to 10 μM, 2 to 20 μM, 2 to 15 μM, 2 to 10 μM, 3 to 20 μM, 3 to 15 μM, 3 to 10 μM, 4 to 20 μM, 4 to 15 μM, 4 to 10 μM, 4 to 8 μM, 4 to 6 μM, 5 to 20 μM, 5 to 15 μM, or 5 to 10 μM, for example, at a concentration of 5 μM or 10 μM.

[0044] The composition may comprise a METTL3 inhibitor at a concentration of 1 to 20 μM, 1 to 15 μM, 1 to 10 μM, 2 to 20 μM, 2 to 15 μM, 2 to 10 μM, 3 to 20 μM, 3 to 15 μM, 3 to 10 μM, 4 to 20 μM, 4 to 15 μM, 4 to 10 μM, 4 to 8 μM, 4 to 6 μM, 5 to 20 μM, 5 to 15 μM, or 5 to 10 μM, for example, at a concentration of 5 μM or 10 μM.

[0045] In the present invention, the cancer may be at least one selected from the group consisting of leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer.

[0046] In one embodiment of the present invention, the leukemia is acute lymphoblastic leukemia, B cell precursor acute lymphoblastic leukemia, T cell precursor acute lymphoblastic leukemia, acute biphenotypic leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, B cell prolymphocytic leukemia, acute myelogenous leukemia, acute promyelocytic leukemia, acute myeloblastic leukemia, acute megakaryoblastic leukemia, chronic myelogenous leukemia The disease may be at least one selected from the group consisting of, but is not necessarily limited to, chronic myelomonocytic leukemia, hairy cell leukemia, T cell prolymphocytic leukemia, large granular lymphocytic leukemia, adult T cell leukemia, and chronic eosinophilic leukemia.

[0047] In one embodiment of the present invention, the lymphoma may be at least one T-cell lymphoma (TCL) selected from the group consisting of T-cell non-Hodgkin lymphoma, cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL), anaplastic large cell lymphoma (ALCL), adult T-cell leukemia / lymphoma (ATLL), angioimmunoblastic T-cell lymphoma (AITL), extranodal NK / T-cell lymphoma, and hepatosplenic T-cell lymphoma, but is not necessarily limited thereto.

[0048] In one embodiment of the present invention, the lymphoma is Burkitt's lymphoma, B cell non-Hodgkin lymphoma (NHL), B cell chronic lymphocytic leukemia (B-CLL), B cell acute lymphoblastic leukemia (B-ALL), large B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, marginal zone B-cell lymphoma, chronic lymphocytic leukemia (CLL), mantle cell lymphoma, primary mediastinal large B-cell lymphoma (PMBCL), intravascular large B cell lymphoma (IVLBCL), It may be one or more types of B-cell lymphoma (BCL) selected from the group consisting of, but not necessarily limited to, primary effusion lymphoma, lymphomatoid granulomatosis, primary central nervous system lymphoma (PCNSL), ALK-positive large B-cell lymphoma, and plasmablastic lymphoma (PBL).

[0049] In one embodiment of the present invention, the lymphoma may be nodular lymphocyte predominant Hodgkin lymphoma, lymphocyte rich classical Hodgkin lymphoma, nodular sclerosis classical Hodgkin lymphoma, mixed cellularity classical Hodgkin lymphoma, or lymphocyte depletion classical Hodgkin lymphoma.

[0050] In another embodiment of the present invention, the lymphoma may be non-Hodgkin's lymphoma that occurs in a human immunodeficiency virus (HIV) patient, an organ transplant recipient, a patient who has received stem cell transplantation and high-dose chemotherapy, a hereditary immunodeficiency syndrome, or an autoimmune disease patient.

[0051] In another embodiment of the present invention, the lymphoma may be Burkitt's lymphoma, nasopharyngeal carcinoma (NPC), or Hodgkin's disease that occurs in an immunocompromised patient infected with Ebstein-Barr Virus (EBV).

[0052] In another embodiment of the present invention, the lymphoma may be gastric mucosa-associated lymphoid tissue lymphoma caused by Helicobacter pylori infection.

[0053] In one embodiment of the present invention, the lung cancer may be, but is not necessarily limited to, non-small cell lung cancer, small cell lung cancer, pancoast tumor, or lung carcinoid tumor.

[0054] In one embodiment of the present invention, the non-small cell lung cancer may be, but is not necessarily limited to, squamous cell carcinoma, adenocarcinoma, or large-cell carcinoma.

[0055] In one embodiment of the present invention, the pancreatic cancer may be at least one pancreatic cystic tumor selected from the group consisting of serous cystadenoma, mucinous cystic neoplasm, intraductal papillary mucinous neoplasm (IPMN), and solid pseudopapillary tumor, or at least one malignant pancreatic tumor selected from the group consisting of pancreatic ductal adenocarcinoma, acinar cell carcinoma, and neuroendocrine tumor, but is not necessarily limited thereto.

[0056] In one embodiment of the present invention, the breast cancer may be at least one selected from the group consisting of triple negative breast cancer (TNBC), metastatic breast cancer, refractory breast cancer, androgen receptor (AR)-positive breast cancer, estrogen receptor (ER)-positive breast cancer, and receptor tyrosine-protein kinase erbB-2 (HER2)-positive breast cancer, or may be breast cancer that has failed treatment with at least one therapeutic agent selected from the group consisting of tamoxifen, toremifene, exemestane, anastrozole, letrozole, trastuzumab, ado-trastuzumab, emtansine, pertuzumab, lapatinib, bevacizumab, and fulvestrant.

[0057] The pharmaceutical composition of the present invention may additionally comprise a pharmaceutically acceptable carrier.

[0058] The term "pharmaceutically acceptable" as used herein means, as commonly used in the pharmaceutical field, that when administered, it does not stimulate the organism and does not inhibit the biological activity and properties of the administered compound.

[0059] In the present invention, any carrier commonly used in the art may be used as the carrier. Non-limiting examples of the carrier include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, maltodextrin, glycerol, ethanol, or combinations thereof.

[0060] The pharmaceutical composition of the present invention may be used by adding other pharmaceutically acceptable additives, such as excipients, diluents, antioxidants, buffers, or bacteriostatic agents, if necessary, and may be used by additionally adding fillers, bulking agents, wetting agents, disintegrating agents, dispersing agents, surfactants, binders, or lubricants.

[0061] The pharmaceutical composition of the present invention may be administered in various dosages depending on factors such as the formulation method, administration method, patient age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. The dosage may be determined or prescribed as an effective dosage for the desired treatment or prevention. For example, the daily dosage of the pharmaceutical composition of the present invention may be 0.0001-1000 mg / kg.

[0062] The pharmaceutical composition of the present invention can be manufactured in a unit dose form or can be manufactured by placing it in a multi-dose container by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by a person having ordinary skill in the art to which the present invention pertains, and thereby. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, suppository, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer, but is not limited thereto.

[0063] The dosage of the pharmaceutical composition of the present invention may vary depending on the patient's age, weight, sex, dosage form, health condition, and disease severity, and may be administered once or several times a day at regular intervals, depending on the judgment of a doctor or pharmacist. For example, the daily dosage may be 1 to 1000 ug / mL based on the active ingredient content, but this is an example of an average case, and the dosage may be higher or lower depending on individual differences.

[0064] Another aspect of the present invention relates to a combination kit for preventing or treating cancer comprising (i) a BTK degrader and (ii) a METTL3 inhibitor.

[0065] In one embodiment of the present invention, the (i) BTK degrader and (ii) METTL3 inhibitor may be contained in the same container or in different containers.

[0066] The above kit may additionally include not only the active ingredients (i) a BTK degrader and (ii) a METTL3 inhibitor, but also containers, materials, package inserts, etc. commonly used in the art suitable for use as a combination kit for the prevention or treatment of cancer.

[0067] The above containers may include, but are not necessarily limited to, vials, syringes, bottles, etc.

[0068] The above materials may include, but are not necessarily limited to, diluents such as saline solution, filters, IV bags and lines, needles and syringes, etc.

[0069] The package insert may be instructions for using a BTK degrader and the METTL3 inhibitor in combination to treat or delay the progression of cancer, or instructions for treating a subject for cancer using the BTK degrader and the METTL3 inhibitor.

[0070] In the present invention, the BTK decomposer may be MT-802, but is not necessarily limited thereto.

[0071] In the present invention, the METTL3 inhibitor may be STM2457, but is not necessarily limited thereto.

[0072] In the present invention, the (i) BTK decomposition agent and (ii) METTL3 inhibitor may be administered simultaneously or sequentially.

[0073] The term “simultaneously” as used herein means that two preparations or pharmaceutical compositions are administered at the same time.

[0074] The term "sequentially" as used herein means that the active ingredients or pharmaceutical compositions are administered one after the other, rather than simultaneously. For example, "sequentially" administration may mean that one agent / pharmaceutical composition is administered within 1 minute, 5 minutes, 10 minutes, 30 minutes, or several hours, for example, within 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 12 hours, 24 hours, or 48 hours, of another agent / pharmaceutical composition, provided that the circulating half-life of the first agent administered is sufficiently long to allow both agents to be present simultaneously in therapeutically effective amounts, but is not necessarily limited thereto. The time delay in administration between components may vary depending on the exact nature, interactions, and respective half-lives of the components.

[0075] In one embodiment of the present invention, the (i) BTK degrader and (ii) METTL3 inhibitor may be administered at the same time or at different time intervals.

[0076] In one embodiment of the present invention, the (i) BTK decomposer and (ii) METTL3 inhibitor may be administered via the same route of administration or different routes of administration.

[0077] In the present invention, the cancer may be at least one selected from the group consisting of leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer.

[0078] Since the kit of the present invention comprises the active ingredient of the pharmaceutical composition described above, description of duplicate content is omitted to avoid excessive complexity of this specification.

[0079] Another aspect of the present invention relates to a method for preventing or treating cancer by administering to a subject a pharmaceutical composition comprising a BTK decomposer and a METTL3 inhibitor as active ingredients.

[0080] In one embodiment of the present invention, the BTK degrader and the METTL3 inhibitor may be administered simultaneously, sequentially, or alternatingly. When the BTK degrader and the METTL3 inhibitor are administered alternately, each agent may be sequentially administered at a time interval of, but not limited to, about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 12 hours, about 24 hours, about 48 hours, about 72 hours, about 96 hours, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 8 weeks, or about 12 weeks.

[0081] In one embodiment of the present invention, the BTK degrading agent may be administered at a dose of, but is not limited to, 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 50 mg / kg, 1 mg / kg to 1000 mg / kg, 1 mg / kg to 500 mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 5 mg / kg to 1000 mg / kg, 5 mg / kg to 500 mg / kg, 5 mg / kg to 100 mg / kg, or 5 mg / kg to 50 mg / kg.

[0082] In one embodiment of the present invention, the METTL3 inhibitor may be administered at a dose of, but is not limited to, 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 50 mg / kg, 1 mg / kg to 1000 mg / kg, 1 mg / kg to 500 mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 5 mg / kg to 1000 mg / kg, 5 mg / kg to 500 mg / kg, 5 mg / kg to 100 mg / kg, or 5 mg / kg to 50 mg / kg.

[0083] In one embodiment of the present invention, the pharmaceutical composition may be administered 1 to 6 times, 1 to 5 times, 1 to 4 times, or 1 to 3 times daily, as needed, but is not limited thereto.

[0084] In one embodiment of the present invention, the pharmaceutical composition may be in the form of, but is not limited to, tablets, capsules, granules, syrups, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, creams, and injections.

[0085] The term "subject" in this specification may be a mammal including a human, for example, but not limited to, a human, monkey, cow, horse, sheep, pig, cat, dog, mouse, rat, rabbit or guinea pig.

[0086] The term "administration" as used herein means providing a given substance to a subject by any suitable method, and the route of administration of the composition comprising the BTK decomposer and METTL3 inhibitor of the present invention as active ingredients may be administered orally or parenterally through any common route as long as it can reach the target tissue. In addition, the composition of the present invention may be administered using any device capable of delivering the active ingredient to the target cell. For example, the pharmaceutical composition of the present invention may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, topical administration, intranasal administration, intrapulmonary administration, rectal administration, intrathecal administration, ocular administration, skin administration, and transdermal administration, but is not limited thereto.

[0087] Another aspect of the present invention relates to the use of a pharmaceutical composition comprising a BTK decomposer and a METTL3 inhibitor as active ingredients for the prevention or treatment of cancer.

[0088] Another aspect of the present invention relates to a combination therapy use of a BTK degrader and a METTL3 inhibitor for the prevention or treatment of cancer.

[0089] Another aspect of the present invention relates to a method for preventing or treating cancer, comprising administering to a subject in need of treatment an effective amount of a BTK degrader and a METTL3 inhibitor.

[0090] The term “subject” in this specification is meant to include humans (e.g., patients) and mammals (e.g., mice, rats, dogs, cats, rabbits, chickens, monkeys, etc.).

[0091] The term "effective amount" as used herein means an amount (e.g., dose) of a pharmaceutical composition that provides a significant reduction in the clinical symptoms of a disease or condition to be treated without causing excessive toxic side effects.

[0092] The term “dose” as used herein means the weight (e.g., milligrams (mg)) of active substance per kilogram (kg) of body weight of the subject.

[0093] In one embodiment of the present invention, the BTK degrader and the METTL3 inhibitor may be present as separate formulations, but are not limited thereto.

[0094] In one embodiment of the present invention, the BTK degrader and the METTL3 inhibitor may be administered simultaneously, sequentially, or alternately. In one embodiment of the present invention, the BTK degrader and the METTL3 inhibitor may be administered in combination.

[0095] The present invention relates to a pharmaceutical composition for preventing or treating cancer, comprising a BTK degrader and a METTL3 inhibitor. It has been confirmed that when a BTK degrader and a METTL3 inhibitor are administered together based on proteolysis-targeting chimera (PROTAC) technology, there is a synergistic effect in which they exhibit remarkable anticancer activity compared to when they are administered alone, and thus, it can create a preventive or therapeutic effect on various cancers such as leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer.

[0096] Figure 1 is a graph showing the results of measuring the optical density (OD) when a BTK decomposition agent (MT-802) and a METTL3 inhibitor (STM2457) according to one embodiment of the present invention were treated alone or in combination with a T-cell leukemia cell line (Jurkat).

[0097] Figure 2 is a graph showing the results of measuring absorbance when a BTK decomposition agent (MT-802) and a METTL3 inhibitor (STM2457) according to one embodiment of the present invention were treated alone or in combination with a B-cell lymphoma cell line (Daudi).

[0098] FIG. 3 is a graph showing the results of measuring cell survival rate (%) when a BTK decomposition agent (MT-802) and a METTL3 inhibitor (STM2457) according to one embodiment of the present invention were treated alone or in combination with a lung cancer cell line (A549).

[0099] Figure 4 is a graph showing the results of measuring cell survival rate when a BTK decomposition agent (MT-802) and a METTL3 inhibitor (STM2457) according to one embodiment of the present invention were treated alone or in combination with a pancreatic cancer cell line (Capan-1).

[0100] FIG. 5 is a graph showing the results of measuring cell survival rate when a BTK decomposition agent (MT-802) and a METTL3 inhibitor (STM2457) according to one embodiment of the present invention were treated alone or in combination with a breast cancer cell line (MDA-MB-231).

[0101] The present invention relates to a pharmaceutical composition for preventing or treating cancer, comprising a BTK decomposer and a METTL3 inhibitor as active ingredients.

[0102] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only intended to illustrate the present invention, and the scope of the present invention is not limited by these examples.

[0103] Unless otherwise stated, all numbers, values ​​and / or expressions expressing ingredients, reaction conditions and quantities of ingredients used in this specification are approximations that inherently reflect, among other things, the various uncertainties of measurement that arise in obtaining such values ​​and therefore should be understood as being modified in all instances by the term "about", and when a numerical range is disclosed herein, such range is continuous and includes every value from the minimum value to the maximum value inclusive, unless otherwise stated.

[0104] Also, the term "or" is intended to mean an inclusive "or" rather than an exclusive "or." That is, where the connection or use between constructions is not otherwise specified or clear from the context, i.e., if X includes A; X includes B; or X includes both A and B, "X includes A or B" can be applied to any of these cases.

[0105] And, throughout the specification, whenever a part is said to "include" a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise specifically stated.

[0106]

[0107] Example 1: Combination treatment experiment of BTK degrader and METTL3 inhibitor on T-cell leukemia cell lines

[0108] Using Jurkat, a T-cell leukemia cell line, 2×10 per well 4 Cells (100 μl) were cultured in RPMI medium (Gibco) supplemented with 10% FBS (Gibco), antibiotics (Gibco), and glutamax (Gibco). After stimulation with PROTAC-based BTK degrader (MT-802) and METTL3 inhibitor (STM2457) according to the conditions, the cells were cultured in a CO2 incubator for 72 h. Then, CCK-8 (Cell Counting Kit-8, BIOMAX) reagent was stimulated at 10 μl per well, and the optical density (OD) was measured at 450 nm after 3 h. Statistics were analyzed using t-test, and *p<0.05, **p<0.01, ***p<0.001 means.

[0109] Treatment group Absorbance (OD) Control group 1.474 MT-802 alone treatment group (5 μM) 0.646 MT-802 alone treatment group (10 μM) 0.46 275 STM2457 alone treatment group (5 μM) 0.79 STM2457 alone treatment group (10 μM) 0.47 425 MT-802 (5 μM) + STM2457 (5 μM) combination treatment group 0.2625

[0110]

[0111] As can be seen in Fig. 1 and Table 1 above, in the T-cell leukemia cell line (Jurkat), the PROTAC-based BTK degrader MT-802 single treatment group showed an absorbance value of 0.46275 at 10 μM, and the METTL3 inhibitor STM2457 single treatment group showed an absorbance value of 0.47425 at 10 μM, whereas the combination treatment group of MT-802 (5 μM) and STM2457 (5 μM) showed an absorbance value of 0.2625, confirming a statistically significant decrease compared to the single treatment group.

[0112] In particular, the absorbance of the combined treatment group of BTK decomposition agent MT-802 (5 μM) and METTL3 inhibitor STM2457 (5 μM) was significantly lower at 0.2625 than the average value of the absorbance of the single treatment group of BTK decomposition agent MT-802 (10 μM) and the single treatment group of METTL3 inhibitor STM2457 (10 μM) (0.46275 + 0.47425) / 2 = 0.4685, confirming that the combined treatment group of BTK decomposition agent and METTL3 inhibitor had a synergistic effect on the anticancer activity against leukemia compared to the single treatment group.

[0113]

[0114] Example 2: Combination treatment experiment of BTK degrader and METTL3 inhibitor on B-cell lymphoma cell lines

[0115] Using Daudi, a B-cell lymphoma cell line, 2 × 10 per well 4Cells (100 μl) were cultured in RPMI medium (Gibco) supplemented with 10% FBS (Gibco), antibiotics (Gibco), and glutamax (Gibco). After stimulation with PROTAC-based BTK degrader (MT-802) and METTL3 inhibitor (STM2457) according to the conditions, the cells were cultured in a CO2 incubator for 72 h. Then, CCK-8 (Cell Counting Kit-8, BIOMAX) reagent was stimulated at 10 μl per well, and the optical density (OD) was measured at 450 nm after 3 h, and the results are presented. Statistics were analyzed using t-test, and *p<0.05, **p<0.01, ***p<0.001 means significant.

[0116] Treatment group Absorbance (OD) Control group 1.0 19 33 MT-802 alone treatment group (5 μM) 0.68 MT-802 alone treatment group (10 μM) 0.31 6 STM2457 alone treatment group (5 μM) 0.51 5 25 STM2457 alone treatment group (10 μM) 0.32 3 25 MT-802 (5 μM) + STM2457 (5 μM) combination treatment group 0.19 5 25

[0117]

[0118] As can be seen in Fig. 2 and Table 2 above, in the B-cell lymphoma cell line (Daudi), the PROTAC-based BTK degrader MT-802 single treatment group showed an absorbance value of 0.316 at 10 μM, and the METTL3 inhibitor STM2457 single treatment group showed an absorbance value of 0.32325 at 10 μM, whereas the combination treatment group of MT-802 (5 μM) and STM2457 (5 μM) showed an absorbance value of 0.19525, confirming a statistically significant decrease compared to the single treatment group.

[0119] In particular, the absorbance of the combined treatment group of BTK decomposition agent MT-802 (5 μM) and METTL3 inhibitor STM2457 (5 μM) was significantly lower at 0.19525 than the average value of the absorbance of the single treatment group of BTK decomposition agent MT-802 (10 μM) and the single treatment group of METTL3 inhibitor STM2457 (10 μM) (0.316 + 0.32325) / 2 = 0.319625, confirming that the combined treatment group of BTK decomposition agent and METTL3 inhibitor had a synergistic effect on anticancer activity against lymphoma compared to the single treatment group.

[0120]

[0121] Example 3: Experimental study of combined treatment of lung cancer cell lines with a BTK degrader and a METTL3 inhibitor.

[0122] Using lung cancer cell line A549 (A-549), 5.0×10 3 Cells were treated with MT-802, a PROTAC-based BTK degrader (dBTK), and STM2457, a METTL3 inhibitor (iMETTL3), alone or in combination, and cell viability was determined 48 hours later using a CCK-8 assay (Cell Counting Kit-8, BIOMAX). Statistical analysis was performed using a t-test, and *p<0.05, **p<0.01, ***p<0.001 indicates significant differences.

[0123] Treatment group Cell viability (%) Control group 100 DMSO only treatment group 75.35 dBTK only treatment group (5 μM) 74.05 dBTK only treatment group (10 μM) 65.53 iMETTL3 only treatment group (5 μM) 78.27 iMETTL3 only treatment group (5 μM) 72.62 dBTK (5 μM) + iMETTL3 (5 μM) combination treatment group 55.83

[0124]

[0125] As can be seen in FIG. 3 and Table 3 above, the BTK decomposition agent (dBTK) MT-802 single treatment group and the METTL3 inhibitor (iMETTL3) STM2457 single treatment group did not have a significant effect on the survival rate of the A549 cell line, whereas the combined treatment group of the BTK decomposition agent (dBTK) MT-802 and the METTL3 inhibitor (iMETTL3) STM2457 statistically significantly inhibited the survival rate of the A549 cell line.

[0126] In particular, the average cell viability of the BTK decomposition agent MT-802 (10 μM) single treatment group and the METTL3 inhibitor STM2457 (10 μM) single treatment group was (65.53% + 72.62%) / 2 = 69.075%, which is significantly lower than the cell viability of the combined treatment group of the BTK decomposition agent MT-802 (5 μM) and the METTL3 inhibitor STM2457 (5 μM), which was 55.83%. Therefore, it was confirmed that the combined treatment group of the BTK decomposition agent and the METTL3 inhibitor had a synergistic effect on anticancer activity against lung cancer compared to the single treatment group.

[0127]

[0128] Example 4: Combination treatment experiment of BTK degrader and METTL3 inhibitor on pancreatic cancer cell lines.

[0129] Using the pancreatic cancer cell line Capan-1, 5.0×10 3 Cells were treated with MT-802, a PROTAC-based BTK degrader (dBTK), and STM2457, a METTL3 inhibitor (iMETTL3), alone or in combination, and cell viability was determined 48 hours later using a CCK-8 assay (Cell Counting Kit-8, BIOMAX). Statistical analysis was performed using a t-test, and *p<0.05, **p<0.01, ***p<0.001 indicates significant differences.

[0130] Treatment group Cell viability (%) Control group 100.02 DMSO only treatment group 73.77 dBTK only treatment group (5 μM) 86.64 dBTK only treatment group (10 μM) 65.98 iMETTL3 only treatment group (5 μM) 81.38 iMETTL3 only treatment group (10 μM) 74.26 dBTK (5 μM) + iMETTL3 (5 μM) combination treatment group 49.59

[0131]

[0132] As can be seen in Fig. 4 and Table 4 above, the BTK decomposition agent (dBTK) MT-802 single treatment group and the METTL3 inhibitor (iMETTL3) STM2457 single treatment group did not have a significant effect on the survival rate of the Capan-1 cell line, whereas the combined treatment group of the BTK decomposition agent (dBTK) MT-802 and the METTL3 inhibitor (iMETTL3) STM2457 statistically significantly inhibited the survival rate of the Capan-1 cell line.

[0133] In particular, the average cell viability of the BTK decomposition agent MT-802 (10 μM) single treatment group and the METTL3 inhibitor STM2457 (10 μM) single treatment group was (65.98% + 74.26%) / 2 = 70.12%, which is significantly lower than the cell viability of the combined treatment group of the BTK decomposition agent MT-802 (5 μM) and the METTL3 inhibitor STM2457 (5 μM), which was 49.59%. Therefore, it was confirmed that the combined treatment group of the BTK decomposition agent and the METTL3 inhibitor had a synergistic effect on anticancer activity against pancreatic cancer compared to the single treatment group.

[0134]

[0135] Example 5: Experimental study of combined treatment of BTK degraders and METTL3 inhibitors on breast cancer cell lines.

[0136] Using the breast cancer cell line MDA-MB-231, 5.0×10 3Cells were treated with MT-802, a PROTAC-based BTK degrader (dBTK), and STM2457, a METTL3 inhibitor (iMETTL3), alone or in combination, and cell viability was determined 48 hours later using a CCK-8 assay (Cell Counting Kit-8, BIOMAX). Statistical analysis was performed using a t-test, and *p<0.05, **p<0.01, ***p<0.001 indicates significant differences.

[0137] Treatment group Cell viability (%) Control group 100 DMSO only treatment group 70.5 dBTK only treatment group (5 μM) 72.07 dBTK only treatment group (10 μM) 63.67 iMETTL3 only treatment group (5 μM) 69.66 iMETTL3 only treatment group (10 μM) 55.38 dBTK (5 μM) + iMETTL3 (5 μM) combination treatment group 45.05

[0138]

[0139] As can be seen in FIG. 5 and Table 5 above, the survival rate inhibition effect of the MDA-MB-231 cell line was not significant in the group treated with the BTK decomposition agent (dBTK) MT-802 alone and the group treated with the METTL3 inhibitor (iMETTL3) STM2457 alone, whereas the survival rate of the MDA-MB-231 cell line was statistically significantly inhibited in the group treated with the combination of the BTK decomposition agent (dBTK) MT-802 and the METTL3 inhibitor (iMETTL3) STM2457.

[0140] In particular, the average cell viability of the BTK decomposition agent MT-802 (10 μM) single treatment group and the METTL3 inhibitor STM2457 (10 μM) single treatment group was (63.67% + 55.38%) / 2 = 59.5252%, which is significantly lower than the cell viability of the combined treatment group of the BTK decomposition agent MT-802 (5 μM) and the METTL3 inhibitor STM2457 (5 μM), which was 45.05%. Therefore, it was confirmed that the combined treatment group of the BTK decomposition agent and the METTL3 inhibitor had a synergistic effect on the anticancer activity against breast cancer compared to the single treatment group.

[0141]

[0142] Sintering

[0143] From the above results, the inventors of the present invention confirmed that when a PROTAC-based BTK degrader (MT-802) and a METTL3 inhibitor (STM2457) were combined, compared to when they were treated alone, there was a synergistic effect on anticancer activity that significantly inhibited the growth and cell viability of leukemia (Jurkat), lymphoma (Daudi), lung cancer (A549), pancreatic cancer (Capan-1), and breast cancer (MDA-MB-231) cell lines.

[0144]

[0145] The present invention relates to a pharmaceutical composition for preventing or treating cancer, comprising a BTK degrader and a METTL3 inhibitor. It has been confirmed that when a BTK degrader and a METTL3 inhibitor are administered together based on proteolysis-targeting chimera (PROTAC) technology, there is a synergistic effect in which they exhibit remarkable anticancer activity compared to when they are administered alone, and thus, it can create a preventive or therapeutic effect on various cancers such as leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer.

Claims

1. A pharmaceutical composition for preventing or treating cancer, comprising a BTK decomposer and a METTL3 inhibitor as active ingredients.

2. A pharmaceutical composition according to claim 1, wherein the composition is for combined administration of a BTK decomposer and a METTL3 inhibitor.

3. A pharmaceutical composition according to claim 1, characterized in that the BTK decomposer is based on PROTAC (Proteolysis targeting chimera).

4. A pharmaceutical composition in claim 1, wherein the BTK decomposer is MT-802.

5. A pharmaceutical composition according to claim 1, wherein the METTL3 inhibitor is STM2457.

6. A pharmaceutical composition in claim 1, wherein the molar ratio of the BTK decomposer and the METTL3 inhibitor is 1:10 to 10:

1.

7. A pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises a BTK decomposer at a concentration of 1 to 20 μM and a METTL3 inhibitor at a concentration of 1 to 20 μM.

8. A pharmaceutical composition according to claim 1, wherein the cancer is at least one selected from the group consisting of leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer.

9. A combination kit for the prevention or treatment of cancer comprising (i) a BTK degrader and (ii) a METTL3 inhibitor.

10. A combination kit in clause 9, wherein the BTK decomposer is MT-802.

11. A combination kit according to claim 9, wherein the METTL3 inhibitor is STM2457.

12. A combination kit in claim 9, wherein (i) the BTK decomposer and (ii) the METTL3 inhibitor are administered simultaneously or sequentially.

13. A combination kit according to claim 9, wherein the cancer is at least one selected from the group consisting of leukemia, lymphoma, lung cancer, pancreatic cancer, and breast cancer.

Images