WO2025071306A1 - Pharmaceutical composition for treating lymphoma comprising bcl-2 inhibitor, selective nuclear export protein inhibitor, and elf4a inhibitor - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating cancer in which an elF4A inhibitor and at least one of a BCL-2 inhibitor or a selective export protein (XPO-1) inhibitor are used in a combination regimen. The present invention relates to a pharmaceutical composition for treating cancer, wherein the elF4A inhibitor is one selected from the group consisting of silvestrol, CR-1-3B, zotatifin, rohinitib, didesmethylrocaglamide, and episylvestrol, and used in a combination regimen with at least one of a BCL-2 inhibitor or a selective nuclear export protein inhibitor.

Description

Pharmaceutical composition for treating lymphoma comprising a BCL-2 inhibitor, a selective nuclear export protein inhibitor and an EIF4A inhibitor

The present invention provides a pharmaceutical composition for treating cancer using a BCL-2 inhibitor and an elF4A inhibitor, Selinexor, and a combination of an elF4A inhibitor, a BCL-2 inhibitor, and Selinexor.

Among the types of malignant lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma (DLBCL) is the most common type, and patients with MYC/BCL-2 double-gene abnormality diffuse large B-cell lymphoma (DHL) have a very poor prognosis because the commonly used R-CHOP immunotherapy is not effective. In particular, the MYC oncogene is an important oncogene that increases early tumorigenesis by playing a role in evading various immune checkpoint mechanisms, and is known as an undruggable target that is difficult to attack with drugs that are difficult to directly inhibit. The importance of MYC targeting in diffuse large B-cell lymphoma continues to be emphasized, and given the frequency of lymphoma occurrence, research that can directly or indirectly inhibit the function of MYC is absolutely necessary in Korea as well.

Since it is difficult to attempt direct inhibition targeting MYC, research is being conducted to indirectly target MYC with the concept of synthetic lethality that approaches by blocking the cell signaling circuits with various interactions. In particular, the present invention suggests the possibility of a new anticancer treatment by confirming the cell killing effect by treating B-cell lymphoma cell lines and T-cell lymphoma with venetoclax, a BCL-2 inhibitor, Silvestrol, Zotatifin, Rohinitib, Didesmethylrocaglamide (DDR), and CR-1-31B, which are elF4A inhibitors capable of controlling the expression of MYC with synthetic lethality, and selinexor, a selective nuclear export protein 1 (XPO-1) inhibitor, alone or in combination.

The purpose of the present invention is to provide a composition for treating cancer by using at least one of an elF4A inhibitor, a BCL-2 inhibitor, and a selective nuclear export protein inhibitor in combination.

Another object of the present invention is to provide a composition for treating cancer by using at least one of an elF4A inhibitor, a BCL-2 inhibitor, and a selective nuclear export protein inhibitor in a combined administration, either sequentially or discontinuously.

Another object of the present invention is to provide a pharmaceutical composition for treating cancer using a combination of a BCL-2 inhibitor and a selective nuclear export protein 1 (XPO-1) inhibitor.

The technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, in order to provide a thorough understanding of the present invention, various specific details are set forth, such as specific configurations, compositions, and processes. However, certain embodiments may be practiced without one or more of these specific details, or in combination with other known methods and configurations. In other instances, well-known processes and manufacturing techniques are not described in specific detail so as not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, configuration, composition, or characteristic described in connection with the embodiment is included in one or more embodiments of the present invention. Thus, the appearances of "in one embodiment" or "an embodiment" in various places throughout this specification are not necessarily referring to the same embodiment of the present invention. Additionally, the particular features, configurations, compositions, or characteristics may be combined in any suitable manner in one or more embodiments.

The term 'elF4A inhibitor' in this specification refers to a compound or drug that inhibits the activity of eIF4A (Eukaryotic Initiation Factor 4A) protein. elF4A is an RNA helicase that plays an important role in eukaryotic protein translation, and functions to unwind double-stranded mRNA during the process of binding the 5' cap region of mRNA to the ribosome. Therefore, elF4A inhibitors inhibit the activity of elF4A that unwinds double strands of mRNA, thereby interfering with the initiation process of protein synthesis, which prevents cells from producing specific proteins, thereby inhibiting the protein synthesis process that is abnormally activated in cancer cells in particular, and thus can be used to inhibit the growth of cancer cells.

The above elF4A inhibitors may include, but are not limited to, silvestrol, CR-1-3B, zotatipine, rohinitib, didesmethyllocaglamide, and episilvestrol.

The term 'silvestrol' in this specification refers to a natural compound derived from plants, particularly from the Aglaila plant. Silvestrol has anticancer properties and is being studied as a potential treatment for various types of cancer, particularly acute leukemia, lymphoma, and neuroblastoma. Silvestrol acts as an inhibitor of elF4A, inhibiting protein synthesis and thereby inhibiting cancer cell growth.

The term 'CR-1-31B' in this specification refers to a synthetic compound structurally similar to silvestrol, which has anticancer effects. In particular, as an inhibitor of elF4A, it inhibits protein synthesis and plays a role in preventing cancer cell growth. CR-1-31B is an analogue of silvestrol, and was developed with the goal of higher efficacy and lower toxicity.

In this specification, the term 'zotatipine' is an anticancer agent that inhibits protein synthesis as an inhibitor of elF4A and thus inhibits cancer cell growth. Clinical studies are in progress, particularly for various cancers such as breast cancer and non-small cell lung cancer.

The term 'rohinitiv' in this specification refers to an inhibitor of elF4A, which acts to inhibit protein synthesis and thereby inhibit cancer cell growth. In particular, it is a potent and specific elF4A inhibitor that induces apoptosis of acute myeloid leukemia (AML) cell lines, thereby reducing the leukemia burden in AML xenograft models.

The term 'didesmethyllocaglamide' in this specification is a potent elF4A inhibitor with a mechanism similar to silvestrol. Didesmethyllocaglamide inhibits the activity of elF4A and thus interferes with protein synthesis in cancer cells. Its effects are being studied particularly in leukemia and lymphoma, and it has a structurally similar function to silvestrol.

The term 'episilvestrol' in this specification refers to a compound that acts as an elF4A inhibitor, similar to silvestrol. Episilvestrol inhibits the helicase activity of elF4A, thereby blocking protein synthesis and inhibiting the growth of cancer cells. Although it has some structural differences compared to silvestrol, the mechanism is the same and it has an anticancer effect by inhibiting protein synthesis.

The term 'BCL-2 inhibitor' in this specification refers to a compound or drug that inhibits the function of the BCL-2 (B-cell lymphoma 2) protein. The BCL-2 protein is an anti-apoptotic protein that plays a role in inhibiting apoptosis in cells, and is known to be overexpressed particularly in cancer cells. A BCL-2 inhibitor binds to the BCL-2 protein, preventing cancer cells from evading apoptosis any longer.

The above BCL-2 inhibitors may include, but are not limited to, venetoclax, navitoclax, obatoclax, and sonorotoclax.

The term 'venetoclax' in this specification refers to a BCL-2 inhibitor, a drug that binds to the BCL-2 protein and blocks its action, thereby inducing cancer cells to self-destruct. It is mainly used to treat chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), and acute myeloid leukemia (AML).

The term 'sonorotoclax' in this specification refers to a next-generation BCL-2 inhibitor, which is currently being studied for its effectiveness in treating various blood cancers, especially chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL). Sonorotoclax, similar to venetoclax, promotes apoptosis of cancer cells by inhibiting the BCL-2 protein, which enables cancer cells to evade apoptosis. In particular, sonorotoclax is attracting attention as a safe and effective treatment method because initial research results have shown that it may have higher selectivity and efficacy than venetoclax.

The term 'export protein 1 (XPO-1) inhibitor' used herein refers to a compound or drug that inhibits the function of the XPO-1 (Exportin-1) protein. XPO-1 is an important transport protein that transports proteins and RNA from the nucleus to the cytoplasm, and plays an important role in helping cells avoid normal apoptosis by exporting key regulatory proteins, such as tumor suppressor proteins such as p53, from the nucleus to the cytoplasm and inhibiting their function. XPO-1 inhibitors bind to XPO-1 and prevent the nuclear transport of tumor suppressor proteins, thereby inhibiting the survival and proliferation of cancer cells.

The term 'selinexor' in this specification refers to a representative XPO-1 inhibitor that is effective in various types of cancer. Selinexor promotes cancer cell death by inducing nuclear accumulation of tumor suppressor proteins.

In this specification, the term 'cancer' refers to a disease in which cells grow and divide abnormally, and is characterized by uncontrolled cell proliferation. Unlike normal cells that undergo regulated growth and division and die after a certain period of time, cancer cells can evade this process and continue to proliferate, invading surrounding tissues or metastasizing to other organs.

The above cancers may include, but are not limited to, breast cancer, lung cancer, colon cancer, prostate cancer, liver cancer, pancreatic cancer, stomach cancer, kidney cancer, ovarian cancer, cervical cancer, oral cancer, nasopharyngeal cancer, esophageal cancer, myeloma, leukemia, lymphoma, melanoma, brain tumor, neuroblastoma, chondrosarcoma, osteosarcoma, bladder cancer, gallbladder cancer, thyroid cancer, small cell lung cancer, Hodgkin's lymphoma, and non-Hodgkin's lymphoma.

In this specification, the term 'lymphoma' refers to a type of cancer that occurs in the lymphatic system (lymph nodes, lymph vessels, lymph fluid, spleen, etc.), which is part of the immune system. Lymphocytes proliferate abnormally or are transformed to form tumors in the lymph nodes or other parts of the lymphatic system. The cause of lymphoma is often unknown, but some say it is related to viruses such as Epstein-Barr virus and abnormal immune regulation. Congenital or acquired immunodeficiency is also an important risk factor. In particular, patients receiving immunosuppressive treatment after organ transplantation, Sjogren's syndrome, lupus, rheumatoid arthritis, and various autoimmune diseases are related to malignant lymphoma. Symptoms of lymphoma include painless, gradual enlargement of lymph nodes, cold symptoms, fever, weight loss, night sweats, and systemic symptoms. When the bone marrow is invaded, a decrease in blood cells is shown in a blood test, and when the central nervous system, such as the spinal cord and brain, is infiltrated, vomiting and headaches are complained of. In this way, symptoms of malignant lymphoma appear in various ways depending on the site of occurrence. Lymphoma is broadly divided into two types: Hodgkin's lymphoma and non-Hodgkin's lymphoma.

The term 'Hodgkin Lymphoma' in this specification refers to lymphoma in which a specific abnormal lymphocyte, the Reed-Sternberg cell, is characteristically found, and tends to be relatively well treated.

The term 'Non-Hodgkin Lymphoma' in this specification includes various types of lymphoma in which Reed-Sternberg cells are not found, and can arise from B cells and T cells. In particular, it is classified as a very complex disease because it has various subtypes, and accounts for approximately 90% of all lymphomas.

The term 'B-cell lymphoma' in this specification refers to a type of lymphoma that arises from B cells. B-cell lymphoma is the most common subtype of non-Hodgkin's lymphoma and can appear in various forms.

The above B-cell lymphomas may include, but are not limited to, diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia, and mantle cell lymphoma.

The term 'T-cell lymphoma' in this specification refers to a type of lymphoma that arises from T cells, and is one of the subtypes of non-Hodgkin's lymphoma. T cells play an important role in the immune system and have the function of defending the body from infection and disease. When these T cells proliferate abnormally or are transformed, T-cell lymphoma occurs. T-cell lymphoma accounts for about 10-15% of non-Hodgkin's lymphoma, and occurs more rarely than B-cell lymphoma.

The above T-cell lymphomas may include, but are not limited to, peripheral T-cell lymphoma, cutaneous T-cell lymphoma, and adult T-cell lymphoma.

The term 'diffuse large B-cell lymphoma (DLBCL)' in this specification refers to a type of non-Hodgkin's lymphoma derived from B cells, which is characterized by rapid proliferation of abnormal B cells in lymph nodes or other parts of the body to form a mass.

The above diffuse large B-cell lymphoma can be divided into various subtypes according to molecular characteristics, and may include, but is not limited to, germinal center B-cell type (GCB) and activated B-cell type (ABC).

The term 'MYC/BCL-2 double-hit diffuse large B-cell lymphoma (DHL)' used in this specification refers to a subtype of diffuse large B-cell lymphoma, which is a lymphoma in which mutations occur simultaneously in the MYC and BCL-2 genes. This double-hit lymphoma is caused by the translocation or overexpression of two genes (MYC and BCL-2) that promote the survival and proliferation of tumor cells, and exhibits more aggressive characteristics than typical diffuse large B-cell lymphoma (DLBCL).

In this specification, the term 'combination use' refers to a method of treating a disease by using two or more drugs or treatments simultaneously. It is used when a greater therapeutic effect is expected, such as showing a synergistic effect, reducing side effects, or suppressing resistance, compared to when each drug is used alone.

In this specification, the term 'apoptosis' refers to the process in which a cell ceases function and dies for a specific reason, and there are two main forms: apoptosis and necrosis.

The term 'apoptosis' in this specification refers to the process by which cells die spontaneously in a programmed manner, and is an important mechanism for maintaining homeostasis in the body.

In this specification, the term 'synergism' means that when two or more elements are used together, the effect is greater than the effect of each element. In other words, when used in combination, it means that the effect of each individual element is increased beyond the simple addition.

In this specification, the term 'additive effect' means that when two or more elements are used together, the same result is produced as when the effects of each are simply added together. In this case, the two elements do not affect each other and the effects are expressed independently.

The term 'agent that reduces the expression level of a gene encoding elF4A protein' used herein may include, but is not limited to, any one or more selected from the group consisting of an antisense nucleotide, a short interfering RNA (siRNA), a short hairpin RNA, and a ribozyme that complementarily bind to a gene encoding said protein, preferably a portion thereof, and may include any and all means that directly or indirectly act on a gene encoding a target elF4A protein to induce an effect of inhibiting its expression, as long as it can be easily derived by a known technique using a method commonly used in the art.

In this specification, the term "EoB (Excess Over Bliss)" refers to a concept for evaluating the synergistic effect between two drugs or compounds, and is a value indicating how much the combined effect of the two drugs exceeds the effect predicted by the Bliss model. When the actual effect is greater than the effect predicted by the Bliss model, the EoB value is positive, indicating a synergistic effect. Conversely, when the actual effect is smaller than the effect predicted by the Bliss model, the EoB value is negative, indicating antagonism. When the EoB value is 0, the effect predicted by the Bliss model and the actual effect match, indicating that the two drugs act independently of each other.

The term "Bliss model" in this specification refers to a statistical model used to explain drug interactions, which is a model that assumes that two or more drugs act independently. This model is used to predict the combined effect when two or more drugs act independently of each other.

In the present invention, the pharmaceutical composition may be characterized as being in the form of a capsule, tablet, granule, injection, ointment, powder or beverage, and the pharmaceutical composition may be characterized as being intended for humans.

In the present invention, the pharmaceutical composition is not limited thereto, but may be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, and aqueous suspensions, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, and fragrances for oral administration, and buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, and the like may be mixed and used for injections, and bases, excipients, lubricants, and preservatives may be used for topical administration. The formulation of the pharmaceutical composition of the present invention may be variously prepared by mixing with the pharmaceutically acceptable carriers described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injections, it can be manufactured in the form of unit dose ampoules or multiple doses. In addition, it can be formulated in the form of solutions, suspensions, tablets, capsules, sustained-release preparations, etc.

Meanwhile, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate or mineral oil. In addition, fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be additionally included.

The routes of administration of the pharmaceutical composition of the present invention include, but are not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal. Oral or parenteral administration is preferred.

The "parenteral" of the present invention includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. Preferably, the pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration, but is not limited thereto.

The pharmaceutical composition of the present invention may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, dosage, administration time, administration route, excretion rate, drug combination, and the severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route, and period, but may be appropriately selected by those skilled in the art, and may be administered at 0.0001 to 50 mg/kg or 0.001 to 50 mg/kg per day. Administration may be administered once a day or divided into several times. The dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition according to the present invention can be formulated as a pill, a dragee, a capsule, a solution, a gel, a syrup, a slurry, or a suspension.

In one specific embodiment of the present invention, a pharmaceutical composition for treating cancer is provided, wherein at least one of an elF4A inhibitor, a BCL-2 inhibitor, and a selective nuclear export protein (XPO-1) inhibitor is used in combination, wherein the nuclear export protein inhibitor is Selinexor, and the pharmaceutical composition for treating cancer is provided, wherein the BCL-2 inhibitor is one selected from the group consisting of Venetoclax, Navitoclax, Obatoclax, and Sonorotoclax, and the pharmaceutical composition for treating cancer is provided, wherein the elF4A inhibitor is Silvestrol, CR-1-3B, Zotatifin, Rohinitib, Didesmethylrocaglamide (DDR), and A pharmaceutical composition for treating cancer is provided, wherein the pharmaceutical composition comprises one selected from the group consisting of episilvestrol.

In one specific embodiment of the present invention, the elF4A inhibitor is an agent that reduces the activity or expression level of the elF4A protein; or an agent that reduces the expression level of a gene encoding the protein, and a pharmaceutical composition for treating cancer is provided, wherein the agent that reduces the expression level of the gene is at least one selected from the group consisting of antisense nucleotides, small interfering RNAs (siRNAs), and ribozymes that complementarily bind to the elF4A gene or a portion thereof.

In one specific embodiment of the present invention, the pharmaceutical composition for treating cancer is provided, wherein at least one of an elF4A inhibitor, a BCL-2 inhibitor, and a selective nuclear export protein inhibitor is used in a combination regimen, either sequentially or discontinuously.

In one specific embodiment of the present invention, the cancer is lymphoma, the pharmaceutical composition is provided, the lymphoma comprises B-cell lymphoma or T-cell lymphoma, the B-cell lymphoma comprises diffuse large B-cell lymphoma, and the pharmaceutical composition comprises bigenic diffuse large B-cell lymphoma.

In one specific embodiment of the present invention, a pharmaceutical composition is provided, wherein the elF4A inhibitor, the BCL-2 inhibitor and the selective nuclear export protein inhibitor are contained at 0.0001 μM to 1.0 μM.

In one specific embodiment of the present invention, the pharmaceutical composition provides a pharmaceutical composition that is administered to a patient with lymphoma, wherein the patient with lymphoma includes a B-cell lymphoma patient or a T-cell lymphoma patient.

In one specific embodiment of the present invention, the pharmaceutical composition is provided in the form of an oral formulation of a powder, granules, capsules, tablets, powder or aqueous suspension, an external preparation, a suppository, or a sterile injectable solution.

In one specific embodiment of the present invention, a pharmaceutical composition for treating cancer is provided, wherein a BCL-2 inhibitor and a selective nuclear export protein 1 (XPO-1) inhibitor are used in combination, and the pharmaceutical composition provides a pharmaceutical composition for treating cancer, wherein the BCL-2 inhibitor and the selective nuclear export protein 1 (XPO-1) inhibitor are used in combination, either sequentially or non-sequentially.

In one specific embodiment of the present invention, a pharmaceutical composition is provided, wherein the BCL-2 inhibitor is any one selected from the group consisting of venetoclax, navitoclax, obatoclax, and sonorotoclax, and the selective nuclear export protein inhibitor is selinexor.

Unless otherwise specifically defined herein, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs.

The present invention confirms that when a BCL-2 inhibitor and an elF4A inhibitor are used in combination, when a nuclear export protein inhibitor selinexor and an elF4A inhibitor are used in combination, and when a BCL-2 inhibitor and selinexor are used in combination, a synergistic effect of high cell death is exhibited in B-cell lymphoma.

The present invention confirms that when Selinexor and an elF4A inhibitor are used in combination, a synergistic effect of high cell death is exhibited in T-cell lymphoma.

In addition, it should be understood that the effects of the present invention are not limited to the effects described above, but include all effects that can be inferred from the composition of the invention described in the detailed description or claims of the present invention.

Figure 1 shows the results of cell death after combined treatment with venetoclax and elF4A inhibitors (silvestrol and CR-1-31B) in OCI-LY7, OCI-LY8, and OCI-LY18 cell lines.

Figure 2 shows the results of cell death after combined treatment with venetoclax and selinexor in OCI-LY1, OCI-LY7, OCI-LY8, OCI-LY18, SUDHL6, and Toledo cell lines.

Figure 3 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and silvestrol in OCI-LY7, OCI-LY8, and OCI-LY18 cell lines.

Figure 4 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and CR-1-31B in OCI-LY7, OCI-LY8, and OCI-LY18 cell lines.

Figure 5 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and selinexor in OCI-LY7, OCI-LY8, and OCI-LY18 cell lines.

Figure 6 is a drawing showing the results of cell death in the HuT-78 cell line after combined treatment with selinexor and CR-1-31B.

Figure 7 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and CR-1-31B in the HuT-78 cell line.

Figure 8 shows the results of cell death after combined treatment with selinexor and CR-1-31B in H9 cell line.

Figure 9 is a graph showing statistical analysis of the results of cell death after combined treatment with Selinexor and CR-1-31B in the H9 cell line.

Figure 10 is a drawing showing the results of cell death after combined treatment with venetoclax and zotatipine in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 11 is a drawing showing the results of cell death after combined treatment with venetoclax and rohinitib in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 12 is a drawing showing the results of cell death after combined treatment with venetoclax and DDR in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 13 is a drawing showing the results of cell death after combined treatment with venetoclax and episilbestrol in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 14 is a drawing showing the results of cell death after combined treatment with venetoclax and CR-1-13B in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 15 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and zotatipine in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 16 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and rohinitib in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 17 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and DDR in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 18 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and episilbestrol in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 19 is a graph showing statistical analysis of the results of cell death after combined treatment with venetoclax and CR-1-13B in OCI-LY1, OCI-LY18, and SU-DHL-6 cell lines.

Figure 20 is a drawing showing the results of cell death after combined treatment with selinexor and zotatipine in OCL-LY1 and OCL-LY18 cell lines.

Figure 21 is a drawing showing the results of cell death after combined treatment with selinexor and rohinitib in OCL-LY1 and OCL-LY18 cell lines.

Figure 22 is a drawing showing the results of cell death after combined treatment with selinexor and DDR in OCL-LY1 and OCL-LY18 cell lines.

Figure 23 is a drawing showing the results of cell death after combined treatment with selinexor and episilbestrol in OCL-LY1 and OCL-LY18 cell lines.

Figure 24 is a drawing showing the results of cell death after combined treatment with selinexor and CR-1-13B in OCL-LY1 and OCL-LY18 cell lines.

Figure 25 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and zotatipine in OCL-LY1 and OCL-LY18 cell lines.

Figure 26 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and rohinitib in OCL-LY1 and OCL-LY18 cell lines.

Figure 27 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and DDR in OCL-LY1 and OCL-LY18 cell lines.

Figure 28 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and episilbestrol in OCL-LY1 and OCL-LY18 cell lines.

Figure 29 is a graph showing statistical analysis of the results of cell death after combined treatment with Selinexor and CR-1-13B in OCL-LY1 and OCL-LY18 cell lines.

Figure 30 is a drawing showing the results of cell death after combined treatment with selinexor and zotatipine in HuT-78 and H9 cell lines.

[Correction under Rule 91 11.11.2024]
Figure 31 is a drawing showing the results of combined treatment with selinexor and rohinitiv in the HuT-78 cell line.

[Correction under Rule 91 11.11.2024]
Figure 32 is a drawing showing the results of cell death caused by combined treatment with selinexor and rohinitiv in H9 cell line.

Figure 33 is a drawing showing the results of cell death caused by combined treatment with selinexor and episilbestrol in HuT-78 and H9 cell lines.

Figure 34 is a drawing showing the results of cell death after combined treatment with selinexor and CR-1-13B in HuT-78 and H9 cell lines.

Figure 35 is a graph showing statistical analysis of the results of cell death caused by combined treatment with selinexor and zotatipine in HuT-78 and H9 cell lines.

Figure 36 is a graph showing statistical analysis of the results of combined treatment with selinexor and rohinitiv in HuT-78 and H9 cell lines.

Figure 37 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and episilbestrol in HuT-78 and H9 cell lines.

Figure 38 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and episilbestrol in HuT-78 and H9 cell lines.

Figure 39 is a graph showing statistical analysis of the results of cell death after combined treatment with selinexor and CR-1-13B in HuT-78 and H9 cell lines.

Hereinafter, the present invention will be described in more detail through examples. These examples are only intended to explain the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention.

Preparation Example. Cell line preparation

The cell lines used in this experiment are as shown in Table 1 below, and each cell culture was cultured in the specified medium as shown in Table 1.

Before drug treatment, 1X10^5/ml cells were dispensed into each well of a 12-well plate (1ml each). Venetoclax was prepared at 0.0025μM, 0.005μM, 0.01μM, 0.02μM, selinexor at 0.05μM, 0.1μM, 0.2μM, 0.5μM, CR-1-31B at 0.125nM, 0.25nM, 0.5nM, 1nM, and zotatipine, rohinitib, didesmethyllocaglamide, and episilbestrol at 1nM, 2nM, 5nM, 10nM, respectively, and each well was treated alone or in combination. Information on the cell lines and subtypes was as shown in Table 1 below.

Cell line Subtype
badge OCI-LY-10 DLBCL ABC IMDM (10% FBS, 1% PenStrep) SU-DHL-2 DLBCL ABC RPMI (10% FBS, 1% PenStrep) HBL-1 DLBCL ABC RPMI (10% FBS, 1% PenStrep) OCI-LY-1 DLBCL GCB/DHL IMDM (20% FBS, 1% PenStrep) OCI-LY-8 DLBCL GCB/DHL RPMI (10% FBS, 1% PenStrep) OCI-LY-18 DLBCL GCB/DHL RPMI (10% FBS, 1% PenStrep) SU-DHL-6 DLBCL GCB/DHL RPMI (10% FBS, 1% PenStrep) Toledo DLBCL GCB/DHL RPMI (10% FBS, 1% PenStrep) OCI-LY-7 DLBCL GCB IMDM (10% FBS, 1% PenStrep) SU-DHL-10 DLBCL GCB RPMI (10% FBS, 1% PenStrep) HuT-78 T cell lymphoma IMDM (10% FBS, 1% PenStrep) H9 T cell lymphoma RPMI (10% FBS, 1% PenStrep)

Example 1. Confirmation of the apoptotic effect by combined treatment with BCL-2 inhibitor and elF4A inhibitor in diffuse large B-cell lymphoma cell line

The combined effect of BCL-2 inhibitors and elF4A inhibitors was confirmed in a broad range of large B-cell lymphoma cell lines.

Specifically, venetoclax was prepared at concentrations of 0.0025 μM, 0.005 μM, 0.01 μM, and 0.02 μM in the cultured diffuse large B-cell lymphoma cell line in Table 1, silvestrol, an eF4A inhibitor, was prepared at concentrations of 0.0025 μM, 0.005 μM, 0.01 μM, and 0.02 μM, CR-1-31B, an eF4A inhibitor, was prepared at concentrations of 0.125 nM, 0.25 nM, 0.5 nM, and 1 nM, and zotatipine, rohinitib, didesmethyllocaglamide, and episilbestrol, respectively, were prepared at concentrations of 1 nM, 2 nM, 5 nM, and 10 nM, respectively, and treated alone or in combination in each well. Afterwards, the cells were cultured in a 37℃, 5% _CO2 incubator for 48 hours and then apoptosis was measured. After 48 hours, the cells were harvested and stained with 7-AAD and Annexin-V, and the apoptosis effect was confirmed using a flow cytometer (Fluorescence-Activated Cell Sorting, FACS).

Example 2. Confirmation of the apoptotic effect by combined treatment with XPO-1 inhibitor Selinexor and elF4A inhibitor in diffuse large B-cell lymphoma cell lines

The combined administration effect of XPO-1 inhibitor and elF4A inhibitor was confirmed in diffuse large B-cell lymphoma cell lines.

Specifically, selinexor was prepared at 0.05 μM, 0.1 μM, 0.2 μM, and 0.5 μM for the cultured diffuse large B-cell lymphoma cell line in Table 1, silvestrol, an eF4A inhibitor, was prepared at 0.0025 μM, 0.005 μM, 0.01 μM, and 0.02 μM, CR-1-31B, an eF4A inhibitor, was prepared at 0.125 nM, 0.25 nM, 0.5 nM, and 1 nM, and zotatipine, rohinitib, didesmethyllocaglamide, and episilbestrol, which are similarly prepared at 1 nM, 2 nM, 5 nM, and 10 nM, respectively, and treated alone or in combination in each well. Afterwards, the cells were cultured in a 37℃, 5% _CO2 incubator for 48 hours and then apoptosis was measured. After 48 hours, the cells were harvested and stained with 7-AAD and Annexin-V and analyzed using a flow cytometer (Fluorescence-Activated Cell Sorting, FACS).

Example 3. Confirmation of the apoptotic effect by combined treatment with BCL-2 inhibitor and XPO-1 inhibitor, Selinexor, in diffuse large B-cell lymphoma cell lines

The combined administration effect of selinexor, a BCL-2 inhibitor and an XPO-1 inhibitor, was confirmed in diffuse large B-cell lymphoma cell lines.

Specifically, venetoclax was prepared at concentrations of 0.0025 μM, 0.005 μM, 0.01 μM, and 0.02 μM, and selinexor was prepared at concentrations of 0.05 μM, 0.1 μM, 0.2 μM, and 0.5 μM, and each well was treated alone or in combination. After that, the cells were cultured in an incubator at 37°C and 5% CO ₂ for 48 hours and then apoptosis was measured. After 48 hours, the cells were harvested and the apoptotic effect was confirmed by staining the harvested cells with 7-AAD and Annexin-V using a flow cytometer (Fluorescence-Activated Cell Sorting, FACS).

Example 4. Confirmation of cell death effect by combined treatment with XPO-1 inhibitor Selinexor and elF4A inhibitor in T cell lymphoma cell lines

The combined administration effect of XPO-1 inhibitor and elF4A inhibitor was confirmed in T-cell lymphoma cell lines.

Specifically, selinexor was prepared at 0.05 μM, 0.1 μM, 0.2 μM, and 0.5 μM for the cultured T-cell lymphoma cell line in Table 1, silvestrol, an elF4A inhibitor, was prepared at 0.0025 μM, 0.005 μM, 0.01 μM, and 0.02 μM, CR-1-31B, an elF4A inhibitor, was prepared at 0.125 nM, 0.25 nM, 0.5 nM, and 1 nM, and zotatipine, rohinitib, didesmethyllocaglamide, and episilbestrol, which are similarly prepared at 1 nM, 2 nM, 5 nM, and 10 nM, respectively, and treated alone or in combination in each well. Afterwards, the cells were cultured in a 37℃, 5% _CO2 incubator for 48 hours and then apoptosis was measured. After 48 hours, the cells were harvested and stained with 7-AAD and Annexin-V and analyzed using a flow cytometer (Fluorescence-Activated Cell Sorting, FACS).

Analysis example. Confirmation and analysis of the results of combined treatment by concentration in each cell line.

Synergistic effects were observed when venetoclax and an elF4A inhibitor were used together, when venetoclax and selinexor were used together, and when selinexor and an elF4A inhibitor were used together in diffuse large B-cell lymphoma cell lines and T-cell lymphoma cell lines. The synergistic effects were expressed through EoB values (Table 2).

Inhibitors and Concentrations Inhibitors and Concentrations Cell line EoB Venetoclax 0.1μM Sylvestrol 10nM OCI-LY7 1.73 0.05μM 25nM OCI-LY8 38.49 0.1μM 25nM OCI-LY18 44.55 Venetoclax 1μM CR-1-31B 0.25nM OCI-LY7 -13.03 0.05μM 0.25nM OCI-LY8 28.90 0.02μM 0.5nM OCI-LY18 29.48 0.02μM 1nM OCI-LY1 42.79 0.02μM 1nM SU-DHL-6 33.94 Venetoclax 0.02μM Steering wheel pin 10nM OCI-LY1 44.27 0.02μM 10nM OCI-LY18 12.99 0.02μM 10nM SU-DHL-6 22.37 Venetoclax 0.02μM Rohinitive 5nM OCI-LY1 18.34 0.02μM 5nM OCI-LY18 12.62 0.02μM 10nM SU-DHL-6 38.43 Venetoclax 0.02μM DDR 5nM OCI-LY1 15.92 0.02μM 10nM OCI-LY18 30.07 0.02μM 5nM SU-DHL-6 14.78 Venetoclax 0.02μM Episilvestrol 5nM OCI-LY1 23.67 0.01μM 10nM OCI-LY18 48.00 0.02μM 5nM SU-DHL-6 20.14 Selinexer 0.1μM Steering wheel pin 2nM OCI-LY1 44.40 0.1μM 5nM OCI-LY18 42.66 0.2μM 5nM HuT-78 46.60 0.2μM 2nM H9 32.38 Selinexer 0.1μM Rohinitive 2nM OCI-LY1 29.76 0.1μM 5nM OCI-LY18 56.46 0.2μM 2nM HuT-78 18.51 0.1μM 2nM H9 38.17 Selinexer 0.1μM DDR 2nM OCI-LY1 37.94 0.1μM 2nM OCI-LY18 62.19 0.1μM 5nM HuT-78 52.36 0.1μM 2nM H9 31.84 Selinexer 0.1μM Episilvestrol 2nM OCI-LY1 22.82 0.1μM 2nM OCI-LY18 59.53 0.1μM 5nM HuT-78 39.78 0.05μM 5nM H9 34.96 Selinexer 0.1μM CR-1-31B 0.25nM OCI-LY1 47.16 0.1μM 0.5nM OCI-LY18 52.04 0.2μM 0.5nM HuT-78 51.59 0.2μM 0.25nM H9 31.49 Venetoclax 1μM Selinexer 0.05μM OCI-LY7 -2.05 0.02μM 0.5μM OCI-LY1 60.00 0.02μM 0.5μM OCI-LY8 47.68 0.01μM 0.05μM OCI-LY18 33.64 0.5μM 0.05μM SU-DHL-6 26.07 0.02μM 0.1μM Toledo 37.41

According to Table 2 above, when venetoclax and silvestrol were used as combination therapy, the EoB value was positive in the double-gene abnormality cell line among B-cell lymphoma, confirming the presence of a synergistic effect. The EoB value was the highest at 44.55 when venetoclax was 0.1 μM and silvestrol was 0.025 nM in the OCI-LY18 cell line, and was 17.78 when venetoclax was 0.05 μM and silvestrol was 0.05 nM. In the OCI-LY8 cell line, the EoB value was the highest at 38.49 when venetoclax was 0.025 μM and silvestrol was 0.025 nM, and was 7.89 when venetoclax was 0.05 μM and silvestrol was 0.05 nM.

When venetoclax and CR-1-31B were used as combination therapy, the EoB value was positive in double-gene abnormality cell lines among B-cell lymphoma, confirming the presence of a synergistic effect. The EoB value was the highest at 40.93 in the OCI-LY18 cell line when venetoclax was 0.01 μM and CR-1-31B was 1 nM, and was 29.48 when venetoclax was 0.02 μM and CR-1-31B was 0.5 nM. In the OCI-LY1 cell line, the EoB value was the highest at 45.27 when venetoclax was 0.01 μM and CR-1-31B was 1 nM, and was the second highest at 42.79 when venetoclax was 0.02 μM and CR-1-31B was 1 nM.

When venetoclax and zotatipine were used in combination therapy, the EoB value was positive in double-gene abnormality cell lines among B-cell lymphoma, confirming the presence of a synergistic effect. The EoB value was the highest at 38.49 when venetoclax was 0.02 μM and zotatipine was 5 nM in the OCI-LY18 cell line, and the second highest at 15.38 when venetoclax was 0.02 μM and zotatipine was 2 nM.

When venetoclax and rohinitib were used in combination therapy, the EoB value was positive in double-gene abnormality cell lines among B-cell lymphoma, confirming the presence of a synergistic effect. The EoB value was the highest at 41.79 when venetoclax was 0.01 μM and rohinitib was 5 nM in the OCI-LY18 cell line, and the second highest at 15.97 when venetoclax was 0.005 μM and rohinitib was 5 nM.

When venetoclax and DDR were used as combination therapy, the EoB value was positive in double-gene abnormality cell lines among B-cell lymphoma, confirming the synergistic effect. The EoB value was the highest at 30.07 when venetoclax was 0.01 μM and DDR was 10 nM in the OCI-LY18 cell line, and the second highest at 28.07 when venetoclax was 0.005 μM and DDR was 10 nM.

When venetoclax and episilbestrol were used as combination therapy, the EoB value was positive in double-gene abnormality cell lines among B-cell lymphoma, confirming the synergistic effect. The EoB value was the highest at 48.00 when venetoclax was 0.01 μM and episilbestrol was 10 nM in the OCI-LY18 cell line, and the second highest at 33.47 when venetoclax was 0.01 μM and episilbestrol was 2 nM.

When selinexor and zotatipine were used in combination therapy, the EoB values were positive in B-cell lymphoma with double genetic abnormalities and T-cell lymphoma cell lines, confirming the synergistic effect. The EoB value was the highest at 63.46 when selinexor was 0.2 μM and zotatipine was 2 nM in the OCI-LY18 cell line, and the second highest at 45.67 when selinexor was 0.1 μM and zotatipine was 2 nM.

When selinexor and rohinitiv were used in combination therapy, the EoB values were positive in B-cell lymphoma with double genetic abnormalities and T-cell lymphoma cell lines, confirming the synergistic effect. The EoB value was the highest at 68.12 when selinexor was 0.1 μM and rohinitiv was 2 nM in the OCI-LY18 cell line, and the second highest at 56.46 when selinexor was 0.1 μM and rohinitiv was 5 nM.

When selinexor and DDR were used as combination therapy, the EoB value was positive in double-gene abnormality cell lines among B-cell lymphoma, confirming the synergistic effect. The EoB value was the highest at 62.19 when selinexor was 0.1 μM and DDR was 2 nM in the OCI-LY18 cell line, and the second highest at 53.92 when selinexor was 0.1 μM and DDR was 5 nM.

When selinexor and CR-1-31B were used in combination therapy, the EoB values were positive in B-cell lymphoma cell lines with double genetic abnormalities and T-cell lymphoma cell lines, confirming a synergistic effect. The EoB value was the highest at 59.53 when selinexor was 0.1 μM and episilbestrol was 2 nM in the OCI-LY18 cell line, and the second highest at 56.89 when selinexor was 0.1 μM and episilbestrol was 5 nM.

When venetoclax and selinexor were used as combination therapy, the EoB value was positive in double-gene abnormality cell lines among B-cell lymphoma, confirming the presence of a synergistic effect. The EoB value was the highest at 33.64 when selinexor was 0.05 μM and venetoclax was 0.01 μM in the OCI-LY18 cell line, and the second highest at 26.48 when selinexor was 0.1 μM and venetoclax was 0.01 μM.

The results showing whether there was a synergistic effect depending on the cell line when each inhibitor was used as a combination therapy are as shown in Table 3 below.

Cell line Subtype Venetoclax + elF4A inhibitor Selinexor + elF4A inhibitor Venetoclax + Selinexer OCI-LY-10 DLBCL ABC No effect No effect No effect SU-DHL-2 DLBCL ABC No effect No effect - HBL-1 DLBCL ABC No effect No effect - OCI-LY-1 DLBCL GCB/DHL Synergy effect Synergy effect Synergy effect OCI-LY-8 DLBCL GCB/DHL Synergy effect Synergy effect Synergy effect OCI-LY-18 DLBCL GCB/DHL Synergy effect Synergy effect Synergy effect SU-DHL-6 DLBCL GCB/DHL Synergy effect Synergy effect Synergy effect Toledo DLBCL GCB/DHL Synergy effect Synergy effect Synergy effect OCI-LY-7 DLBCL GCB No effect No effect No effect SU-DHL-10 DLBCL GCB No effect No effect No effect HuT-78 T cell lymphoma No effect Synergy effect No effect H9 T cell lymphoma No effect Synergy effect No effect

Based on the above examples and Tables 1 to 3, the synergy effect according to each combination administration was summarized as in Table 4 below.

elF4A inhibitor BCL-2 inhibitor XPO1 inhibitor Test subject
Cell line Cell lines with synergistic effects Subtypes of cell lines with synergistic effects Sylvestrol Venetoclax OCI-LY7, OCI-LY8, OCI-LY18 OCI-LY8, OCI-LY18 GCB/DHL CR-1-31B Venetoclax OCI-LY1, OCI-LY18, SU-DHL-6,
OCI-LY7, OCI-LY8, OCI-LY18 OCI-LY1, OCI-LY18, SU-DHL-6
OCI-LY8, OCI-LY18 GCB/DHL Steering wheel pin Venetoclax OCI-LY1, OCI-LY18, SU-DHL-6 OCI-LY1, OCI-LY18, SU-DHL-6 GCB/DHL Rohinitive Venetoclax OCI-LY1, OCI-LY18, SU-DHL-6 OCI-LY1, OCI-LY18, SU-DHL-6 GCB/DHL Didesmethylocaglamide Venetoclax OCI-LY1, OCI-LY18, SU-DHL-6 OCI-LY1, OCI-LY18, SU-DHL-6 GCB/DHL Episilvestrol Venetoclax OCI-LY1, OCI-LY18, SU-DHL-6 OCI-LY1, OCI-LY18, SU-DHL-6 GCB/DHL CR-1-31B Selinexer HuT-78, H9, OCI-LY1, OCI-LY18 HuT-78, H9,
OCI-LY1, OCI-LY18 GCB/DHL, T-cell lymphoma cell line Steering wheel pin Selinexer HuT-78, H9,OCI-LY1, OCI-LY18 HuT-78, H9,
OCI-LY1, OCI-LY18 GCB/DHL, T-cell lymphoma cell line Rohinitive Selinexer HuT-78, H9,OCI-LY1, OCI-LY18 HuT-78, H9,
OCI-LY1, OCI-LY18 GCB/DHL, T-cell lymphoma cell line Didesmethylocaglamide Selinexer HuT-78, H9,OCI-LY1, OCI-LY18 HuT-78, H9,
OCI-LY1, OCI-LY18 GCB/DHL, T-cell lymphoma cell line Episilvestrol Selinexer HuT-78, H9,
OCI-LY1, OCI-LY18 HuT-78, H9,
OCI-LY1, OCI-LY18 GCB/DHL, T-cell lymphoma cell line Venetoclax Selinexer HuT-78, H9,
OCI-LY7, OCI-LY8, OCI-LY18 HuT-78, H9,
OCI-LY8, OCI-LY18 GCB/DHL, T-cell lymphoma cell line

GCB/DHL stands for GCB/DHL, a double-gene abnormality B-cell lymphoma cell line.

While the specific parts of the present invention have been described in detail above, it is obvious to those skilled in the art that such specific descriptions are merely preferred implementation examples and that the scope of the present invention is not limited thereto. Accordingly, it will be obvious to those skilled in the art that various modifications and variations are possible within the scope that does not depart from the technical idea of the present invention described in the claims.

Claims (33)

A pharmaceutical composition for treating cancer, comprising a combination of an elF4A inhibitor, a BCL-2 inhibitor, and a selective nuclear export protein 1 (XPO-1) inhibitor. In the first paragraph, A pharmaceutical composition wherein the above nuclear export protein inhibitor is Selinexor. In the first paragraph, A pharmaceutical composition, wherein the BCL-2 inhibitor is any one selected from the group consisting of venetoclax, navitoclax, obatoclax, and sonorotoclax. In the first paragraph, A pharmaceutical composition wherein the elF4A inhibitor is any one selected from the group consisting of Silvestrol, CR-1-3B, Zotatifin, Rohinitib, Didesmethylrocaglamide (DDR), and Episilvestrol. In clause 1 or 4, A pharmaceutical composition comprising as an active ingredient an agent that reduces the activity or expression level of the elF4A protein, or an agent that reduces the expression level of a gene encoding the protein. In clause 5, A composition wherein the agent for reducing the expression level of the above gene is at least one selected from the group consisting of antisense nucleotides, small interfering RNAs (siRNAs), and ribozymes that complementarily bind to the elF4A gene or a portion thereof. In the first paragraph, The above pharmaceutical composition for treating cancer is a pharmaceutical composition for treating cancer that uses at least one of an elF4A inhibitor, a BCL-2 inhibitor, and a selective nuclear export protein inhibitor in a combination regimen, either sequentially or discontinuously. In any one of paragraphs 1 to 7, A pharmaceutical composition wherein the cancer is lymphoma. In Article 8, A pharmaceutical composition wherein the lymphoma comprises B-cell lymphoma or T-cell lymphoma. In Article 9, A pharmaceutical composition, wherein the B-cell lymphoma comprises diffuse large B-cell lymphoma. In Article 10, A pharmaceutical composition, wherein the above diffuse large B-cell lymphoma comprises bigenic diffuse large B-cell lymphoma. In the first paragraph, A pharmaceutical composition comprising the elF4A inhibitor, the BCL-2 inhibitor and the selective nuclear export protein inhibitor at concentrations of 0.0001 μM to 1.0 μM. In the first paragraph, A pharmaceutical composition, wherein the pharmaceutical composition is administered to a patient with lymphoma. A pharmaceutical composition in claim 13, wherein the lymphoma patient includes a B-cell lymphoma patient or a T-cell lymphoma patient. In the first paragraph, The pharmaceutical composition is prepared in the form of an oral formulation of a powder, granules, capsules, tablets, powder or aqueous suspension, an external preparation, a suppository, or a sterile injectable solution. A pharmaceutical composition for treating cancer using a combination regimen of a BCL-2 inhibitor and a selective nuclear export protein 1 (XPO-1) inhibitor. In Article 16, The above pharmaceutical composition is a pharmaceutical composition for treating cancer, which uses a BCL-2 inhibitor and a selective nuclear export protein inhibitor in a combination regimen, either sequentially or non-sequentially. In Article 16, A pharmaceutical composition, wherein the BCL-2 inhibitor is any one selected from the group consisting of venetoclax, navitoclax, obatoclax, and sonorotoclax. In Article 16, A pharmaceutical composition wherein the selective nuclear export protein inhibitor is selinexor. In Article 16, A pharmaceutical composition wherein the cancer is lymphoma. In Article 16, A pharmaceutical composition wherein the lymphoma comprises B-cell lymphoma or T-cell lymphoma. In Article 16, A pharmaceutical composition, wherein the B-cell lymphoma comprises diffuse large B-cell lymphoma. In Article 16, A pharmaceutical composition, wherein the above diffuse large B-cell lymphoma comprises bigenic diffuse large B-cell lymphoma. A method for treating cancer by sequentially or non-sequentially administering in combination at least one of an elF4A inhibitor, a BCL-2 inhibitor, and a selective nuclear export protein (XPO-1) inhibitor. In Article 24, A method of treating cancer wherein the nuclear export protein inhibitor is Selinexor. In Article 24, A method for treating cancer, wherein the BCL-2 inhibitor is any one selected from the group consisting of venetoclax, navitoclax, obatoclax, and sonorotoclax. In Article 24, A method for treating cancer, wherein the elF4A inhibitor is any one selected from the group consisting of Silvestrol, CR-1-3B, Zotatifin, Rohinitib, Didesmethylrocaglamide (DDR), and Episilvestrol. In clause 24 or clause 27, A method for treating cancer, wherein the elF4A inhibitor comprises as an active ingredient an agent that reduces the activity or expression level of the elF4A protein; or an agent that reduces the expression level of a gene encoding the protein. In Article 28, A method for treating cancer, wherein the agent that reduces the expression level of the above gene is at least one selected from the group consisting of antisense nucleotides, small interfering RNAs (siRNAs), and ribozymes that complementarily bind to the elF4A gene or a portion thereof. In any one of Articles 24 to 29, A method of treatment, wherein the cancer comprises lymphoma. In Article 30, A method of treatment, wherein the lymphoma comprises B-cell lymphoma or T-cell lymphoma. In Article 31, A method of treatment, wherein the above B-cell lymphoma includes diffuse large B-cell lymphoma. In Article 32, A method of treating the above diffuse large B-cell lymphoma, wherein the diffuse large B-cell lymphoma includes bigenic diffuse large B-cell lymphoma.

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