Combination administration therapy of poseltinib for preventing or treating lymphoma

Abstract

The present invention relates to a therapy for preventing or treating lymphoma by administering poseltinib in combination with an immunomodulatory drug. In particular, the present invention achieves a synergistic effect on the prevention or treatment of diffuse large B-cell lymphoma and primary central nervous system lymphoma through the co-administration of poseltinib and lenalidomide.

Description

Combination therapy with posseltinib for the prevention or treatment of lymphoma

[0001] The present invention relates to a pharmaceutical composition for preventing or treating lymphoma comprising Poseltinib and an immunomodulatory drug as active ingredients, a pharmaceutical composition for preventing or treating lymphoma comprising Poseltinib as an active ingredient and administered in combination with an immunomodulatory drug, or a combination of Poseltinib and an immunomodulatory drug.

[0002] Specifically, the lymphoma may be diffuse large B-cell lymphoma (DLBCL) and primary central nervous system lymphoma (PCNSL), and the present invention relates to a pharmaceutical composition that exhibits a synergistic effect with an active ingredient in the pharmaceutical composition.

[0003] Lymphoma is a cancer that develops in the lymphatic system, a crucial component of the body's immune system. The lymphatic system, comprised of lymph nodes, lymph vessels, the spleen, thymus, and bone marrow, is responsible for protecting against infections and removing unwanted substances from the body. Lymphoma is caused by the abnormal proliferation of lymphatic cells, primarily lymphocytes, and is broadly categorized into Hodgkin's lymphoma and non-Hodgkin's lymphoma. Non-Hodgkin's lymphoma is further classified into various subtypes, and clinical manifestations and treatment responses vary significantly depending on the site of origin and cell characteristics.

[0004] Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma and is a malignant tumor with an aggressive growth rate. DLBCL originates in lymphocytes called B cells and is characterized by large tumor cells that proliferate rapidly. DLBCL exhibits a variety of genetic mutations and abnormalities, and the disease's characteristics and treatment response can vary from patient to patient. Major symptoms include enlarged lymph nodes, fever, night sweats, and weight loss, and various other symptoms may accompany the disease depending on the site of the lesion.

[0005] Primary central nervous system lymphoma (PCNSL) is a type of non-Hodgkin's lymphoma, a malignant tumor that arises in the central nervous system, including the brain, spinal cord, and eyes. PCNSL is caused by the abnormal proliferation of lymphoid cells, primarily B cells. Due to the specific location of PCNSL, its clinical presentation and treatment strategies differ from those of lymphomas in other areas. PCNSL is relatively rare, but its incidence is increasing in immunosuppressed patients and the elderly. Tumor cells exhibit infiltrative growth within brain tissue and exhibit various genetic mutations and expression abnormalities. Major symptoms include headache, cognitive decline, personality changes, focal neurological deficits (such as paralysis, paresthesia, and blurred vision), and vomiting due to increased intracranial pressure. Symptoms can vary greatly depending on the location and size of the lesion.

[0006] Recently, in the field of lymphoma treatment, the development of targeted therapies is actively underway.

[0007] Poseltinib is a Bruton's tyrosine kinase (BTK) inhibitor that irreversibly binds to the active site of the enzyme BTK, which plays a crucial role in the B-cell receptor signaling pathway. Unlike existing BTK inhibitors, poseltinib acts noncovalently, allowing it to remain effective even when specific resistance mutations develop.

[0008] Meanwhile, lymphoma, especially DLBCL, has complex disease characteristics, making it often difficult to maximize therapeutic efficacy with a single agent. Furthermore, PCNSL develops in the unique environment of the central nervous system, hindering drug penetration due to the presence of the blood-brain barrier (BBB). Therefore, maximizing therapeutic efficacy with a single agent often proves challenging. Cancer cells activate diverse survival and proliferation pathways, and single-target inhibition alone cannot completely block this complex network. Furthermore, acquisition of drug resistance can be a major limitation of monotherapy. In the case of PCNSL, drug safety is also a critical consideration, as it must act on the sensitive brain region.

[0009] Therefore, in the treatment of lymphoma, a strategy is needed to simultaneously attack multiple pathways of cancer cells with drugs of different mechanisms of action by administering multiple drugs in combination and to reduce the possibility of developing drug resistance.

[0010] Accordingly, the present invention aims to provide a pharmaceutical composition and combination that overcomes the limitations of existing treatments and provides improved therapeutic effects to patients with relapsed or refractory DLBCL and patients with relapsed or refractory PCNSL by co-administering poseltiniib and an immunomodulatory drug. In particular, the goal of this combination therapy is to effectively penetrate the blood-brain barrier and enhance the therapeutic effect on cancer cells in the central nervous system. In addition, the present invention provides a pharmaceutical composition comprising poseltiniib and an immunomodulatory drug to maximize the therapeutic effect of DLBCL and PCNSL through a synergistic effect between the drugs.

[0011] [1] In one aspect of the present invention, the present invention relates to a pharmaceutical composition for preventing or treating lymphoma, comprising Poseltinib and an immunomodulatory drug as active ingredients.

[0012] [2] In another aspect of the present invention, the present invention relates to a pharmaceutical composition for preventing or treating lymphoma, comprising poceltinib as an active ingredient, wherein the pharmaceutical composition is administered in combination with an immunomodulatory drug.

[0013] [3] In the above [1] or [2], the immunomodulatory drug may be at least one selected from the group consisting of Lenalidomide, Thalidomide, Pomalidomide, Imiquimod, Rituximab, Mezigdomide, Iberdomide, Golcadomide, ICP-490, GLB-002, GT919, GT929, HP-001, BTX-1188, and Cemsidomide.

[0014] [4] In the above [3], the immunomodulatory drug may be lenalidomide or semsidomide.

[0015] [5] In the above [1], the pharmaceutical composition may further include as an active ingredient at least one selected from the group consisting of Bruton's Tyrosine Kinase Inhibitors (BTKi), immunosuppressants, targeted anticancer agents, immune checkpoint inhibitors, angiogenesis inhibitors, cytokines and cytokine modulators, cancer vaccines, small molecule compound inhibitors, epigenetic modification modulators, antibodies, anti-CD19 antibodies, anti-CD20 antibodies, anti-CD20 / anti-CD3 bispecific antibodies, antibody-drug conjugates, and chimeric antigen receptor T (CAR-T) cell therapy agents.

[0016] [6] In the above [2], the pharmaceutical composition may be administered in combination with at least one selected from the group consisting of Bruton's Tyrosine Kinase Inhibitors (BTKi), immunosuppressants, targeted anticancer agents, immune checkpoint inhibitors, angiogenesis inhibitors, cytokines and cytokine modulators, cancer vaccines, small molecule compound inhibitors, epigenetic modification modulators, antibodies, anti-CD19 antibodies, anti-CD20 antibodies, anti-CD20 / anti-CD3 bispecific antibodies, antibody-drug conjugates, and chimeric antigen receptor T (CAR-T) cell therapy agents.

[0017] [7] In the above [5] or [6], the BTKi may be at least one selected from the group consisting of Ibrutinib, Acalabrutinib, Zanubrutinib, Tirabrutinib, Orelabrutinib, Spebrutinib, Evobrutinib, Vecabrutinib, Pirtobrutinib, and Fenebrutinib.

[0018] [8] In the above [1] or [2], the lymphoma may be non-Hodgkin lymphoma or Hodgkin lymphoma.

[0019] [9] In the above [8], the non-Hodgkin's lymphoma may be any one selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), primary central nervous system lymphoma (PCNSL), follicular lymphoma, primary mediastinal B-cell lymphoma, mantle cell lymphoma, post-transplant lymphoproliferative disorder, marginal zone lymphoma, Waldenstrom's macroglobulinemia, and Burkitt lymphoma.

[0020]

[0010] In the above [8], the non-Hodgkin's lymphoma may be diffuse large B-cell lymphoma (DLBCL) or primary central nervous system lymphoma (PCNSL).

[0021]

[0011] In the above [4], the posseltinib and lenalidomide can be administered at a concentration ratio of 10:1 to 1:100.

[0022]

[0012] In the above

[0011] , the posseltinib and lenalidomide can be administered at a concentration ratio of 10:1 to 1:1.

[0023]

[0013] In the above [1] or [2], the effective ingredients may be administered simultaneously or sequentially.

[0024]

[0014] In the above [1] or [2], the composition may have a synergistic effect on the prevention or treatment of lymphoma.

[0025]

[0015] In one aspect of the present invention, the present invention relates to a method for preventing or treating lymphoma, comprising administering to a subject a therapeutically effective amount of a pharmaceutical composition comprising poceltinib and an immunomodulatory drug as active ingredients.

[0026]

[0016] In one aspect of the present invention, the present invention relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of poceltinib and an immunomodulatory drug as active ingredients for the prevention or treatment of lymphoma.

[0027]

[0017] In one aspect of the present invention, the present invention relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of poceltinib and an immunomodulatory drug as active ingredients for preparing a medicament for the prevention or treatment of lymphoma.

[0028] The present invention provides a pharmaceutical composition and combination comprising poseltiniib and an immunomodulatory drug as active ingredients for preventing or treating lymphoma, particularly diffuse large B-cell lymphoma (DLBCL) and primary central nervous system lymphoma (PCNSL), thereby improving the therapeutic effect by effectively inhibiting various survival and proliferation pathways of cancer cells through combined administration of poseltiniib and an immunomodulatory drug.

[0029] Figure 1 depicts the effect of single treatments of posseltinib, lenalidomide, and ibrutinib on cell viability in non-Hodgkin's lymphoma cell lines OCI-LY-19, SU-DHL-5, and TMD8.

[0030] Figure 2 shows the cell viability when the TMD8 cell line was treated with posseltinib and ibrutinib at concentrations ranging from 0 uM to 10 uM, respectively, to achieve a 1:1 posseltinib-lenalidomide and ibrutinib-lenalidomide concentration ratio.

[0031] Figure 3 shows the synergism index when posseltinib-lenalidomide and ibrutinib-lenalidomide were combined in a 1:1 ratio and treated with TMD8 cell lines.

[0032] Figure 4 shows the cell viability when posseltinib was treated at a concentration of 0 uM to 0.4 uM and lenalidomide at a concentration of 0 uM to 20 uM to achieve a posseltinib-lenalidomide concentration ratio of 1:50. Also, the cell viability is shown when ibrutinib was treated at a concentration of 0 uM to 0.02 uM and lenalidomide at a concentration of 0 uM to 20 uM to achieve an ibrutinib-lenalidomide concentration ratio of 1:1000.

[0033] Figure 5 shows the synergy index when posseltinib-lenalidomide and ibrutinib-lenalidomide were combined at a ratio of 1:50 and 1:1000, respectively, and treated with TMD8 cell lines.

[0034] Figure 6 shows the cell viability when the OCI-LY19 cell line was treated with posseltinib and ibrutinib at concentrations ranging from 0 uM to 10 uM, respectively, to achieve a 1:1 posseltinib-lenalidomide and ibrutinib-lenalidomide concentration ratio.

[0035] Figure 7 shows the synergy index when posseltinib-lenalidomide and ibrutinib-lenalidomide were combined in a 1:1 ratio and treated with the OCI-LY19 cell line.

[0036] Figure 8 shows the cell viability when SU-DHL-5 cells were treated with poseltiniib and ibrutinib at concentrations ranging from 0 uM to 10 uM, respectively, to achieve a 1:1 poseltiniib-lenalidomide and ibrutinib-lenalidomide concentration ratio.

[0037] Figure 9 shows the synergy index when posseltinib-lenalidomide and ibrutinib-lenalidomide were combined in a 1:1 ratio and treated with SU-DHL-5 cell line.

[0038] Figure 10 shows the results of observing the change in body weight of each animal when vehicle, posseltinib alone, lenalidomide alone, and posseltinib-lenalidomide combination were administered to a mouse Xenograft model.

[0039] Figure 11 shows the results of observing the change in tumor volume in each animal when vehicle, posseltinib alone, lenalidomide alone, and posseltinib-lenalidomide combination were administered to a mouse Xenograft model.

[0040] Figure 12 shows the cell viability and synergy index when posseltinib-cemcidomide was co-administered to the SU-DHL-5 cell line at concentrations of 0 μM to 8 μM of posseltinib and 0 μM to 0.1 μM of cemcidomide.

[0041] Hereinafter, the present invention will be described in more detail.

[0042] Each description and embodiment disclosed in this invention can also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this invention fall within the scope of this invention. Furthermore, the scope of this invention is not limited by the specific descriptions described below.

[0043] Furthermore, those skilled in the art will recognize or be able to ascertain, through routine experimentation, numerous equivalents to the specific embodiments of the invention described herein. Furthermore, such equivalents are intended to be encompassed by the present invention.

[0044]

[0045] definition

[0046] As used herein, the term “consisting of” means that the proportion of a particular component(s) totals 100%. The components or features listed below the term “consisting of” may be essential or mandatory.

[0047] As used herein, the term "comprising" means the presence of a feature, step, or component described below, and does not exclude the presence or addition of one or more features, steps, or components. Components or features described below "comprising" in this specification may be essential or mandatory, but some embodiments may further include other optional or non-essential components or features.

[0048] In this specification, the term “comprising” may, in some implementations, be modified to refer to “consisting essentially of” or “consisting of.”

[0049] As used herein, the term "immunomodulatory drug" refers to a drug that helps the immune system work more effectively by changing its function. This includes all treatments that increase or decrease the immune response. It refers to a substance or drug that regulates the strength or direction of the immune response in the body, and can be used to treat autoimmune diseases or inflammatory diseases by suppressing excessive or imbalanced immune responses, or conversely, to increase the defense against diseases such as infections or cancer by activating immune function in a state where the immune response is depressed. Specifically, immunosuppressants (e.g., corticosteroids, azathioprine, cyclosporine, tacrolimus), immunopotentiators (e.g., imiquimod, interferon, interleukin, vaccine adjuvants, some anticancer immunotherapies), and Immunomodulatory Imide Drugs; IMiDs (e.g., thalidomide, lenalidomide, pomalidomide, semcidomide, ICP-490, GLB-002, GT919, GT929, HP-001, BTX-1188), CELMoDs (e.g., mezigdomide, iverdomide, golcadomide), cytokines (e.g., interferon alpha, interleukin-2), checkpoint inhibitors (e.g., nivolumab, pembrolizumab, atezolizumab), and rituximab.

[0050] Additionally, the “immunomodulatory drug” mentioned in the present invention can be interpreted in a broad sense and can include various types of immunomodulatory drugs that can be utilized in the treatment of lymphoma.

[0051] In this specification, the term “active ingredient” means a substance that exhibits direct pharmacological activity in the diagnosis, treatment, alleviation, cure or prevention of a disease.

[0052] As used herein, the term "lymphoma" refers to cancer arising in the lymphatic system. Lymphoma is broadly divided into non-Hodgkin lymphoma and Hodgkin lymphoma. Specifically, the lymphoma may be any one selected from the group consisting of, but is not limited to, diffuse large B-cell lymphoma (DLBCL), primary central nervous system lymphoma (PCNSL), follicular lymphoma, primary mediastinal B-cell lymphoma, mantle cell lymphoma, post-transplant lymphoproliferative disorder, marginal zone lymphoma, Waldenstrom's macroglobulinemia, and Burkitt lymphoma.

[0053] As used herein, the term "diffuse large B-cell lymphoma (DLBCL)" refers to the most common subtype of non-Hodgkin lymphoma, a malignant tumor that originates from lymphocytes called B cells. The characteristic appearance of DLBCL is that it is a B cell that is large (giant), has a distinct nucleus, and has abundant cytoplasm. The term "diffuse" refers to the characteristic that the cancer cells grow widely and evenly while destroying the normal structure of the lymph node, meaning that it is not limited to a specific area within the lymph node but infiltrates the entire lymph node.

[0054] As used herein, the term "primary central nervous system lymphoma (PCNSL)" refers to a type of non-Hodgkin lymphoma, a malignant tumor that occurs in the central nervous system, including the brain, spinal cord, and eyes. PCNSL primarily originates from lymphocytes called B cells, and is characterized by abnormal proliferation of tumor cells within the tissues of the central nervous system. The term "primary" means that the lymphoma originated in the central nervous system, rather than having originated elsewhere and then metastasized to the central nervous system. The tumor cells of PCNSL can infiltrate the brain parenchyma to form a mass, or can occur in various parts, such as the meninges, spinal cord, and eyes, and their morphology and growth patterns are very diverse.

[0055] In the present invention, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined according to the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route of administration and the excretion rate, the duration of treatment, factors including concurrently used drugs, and other factors well known in the medical field. The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take all of the above factors into consideration and administer an amount that can achieve the maximum effect with the minimum amount without causing side effects, and this can be easily determined by those skilled in the art.

[0056] In the present invention, the term "administration" refers to introducing a given substance into a subject by any suitable method, and the pharmaceutical composition may be administered via any route as long as it can reach the target tissue. Such administration methods include, but are not limited to, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, and rectal administration. However, since proteins are digested during oral administration, it may be desirable to formulate oral compositions to coat the active agent or protect it from degradation in the stomach. Furthermore, the pharmaceutical composition may be administered by any device that allows the active agent to travel to target cells.

[0057] In the present invention, the terms "combined administration," "combined administration," and "combined use" not only mean simultaneous administration, but also may be a dosage form in which metformin, or a pharmaceutically acceptable salt or solvate thereof, and elserine, or a pharmaceutically acceptable salt or solvate thereof, act together on a subject so that each substance can perform a level equivalent to or higher than its original function. Therefore, when the term "combined administration" is used herein, it should be understood that it refers to simultaneous, separate, sequential, or reverse administration, and the order is unlimited. When the administration is sequential, reverse, or separate, the order of administration is not particularly limited, but the interval between administrations of the secondary components should be such that the beneficial effect of the combination is not lost.

[0058] In the present invention, the term "Bruton's Tyrosine Kinase Inhibitors (BTKi)" refers to drugs that inhibit the activity of Bruton's Tyrosine Kinase (BTK), an enzyme that plays a key role in the B-cell receptor (BCR) signal transduction pathway. BTKi binds to the active site of the BTK enzyme and blocks its function, thereby inhibiting the BCR signal transduction pathway and blocking signals necessary for the growth and survival of cancerous B cells, ultimately inducing the death of cancer cells or inhibiting their proliferation. BTKi has an effect of. They can be classified into the first generation (e.g., ibrutinib) that binds irreversibly to BTK but can also affect other kinases and cause side effects, the second generation (e.g., acalabrutinib, zanubrutinib) that have increased specificity for BTK and thus reduced side effects, and the third generation (e.g., pirtobrutinib) that has a non-covalent reversible binding mode and can be effective in patients who are resistant to existing BTKi.

[0059] In the present invention, the term "combination" refers to the combined administration of posseltinib and an immunomodulatory drug, and can be understood to have the same meaning as "combined use." This also includes, but is not limited to, pharmaceutical compositions and pharmaceutical kits characterized by the combined administration of posseltinib and an immunomodulatory drug.

[0060] In this specification, the term “prevention” means any act of suppressing the onset of clinical symptoms of a disease by administering a pharmaceutical composition according to the present invention.

[0061] In this specification, the term “treatment” means any action by which the clinical symptoms of a disease are improved or beneficially changed by administration of the pharmaceutical composition according to the present invention.

[0062] As used herein, the term “therapeutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined based on factors including the subject’s body weight, sex, age, health status, severity, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field.

[0063] In this specification, the term "synergistic effect" means a phenomenon in which, when two or more drugs are used together, a greater effect is produced than when each drug is used alone, and can be quantified by a synergism index.

[0064] Pharmaceutical compositions and combinations

[0065] In one aspect of the present invention, the present invention relates to a pharmaceutical composition for preventing or treating lymphoma comprising poceltinib and an immunomodulatory drug as active ingredients.

[0066] In another aspect of the present invention, the present invention relates to a pharmaceutical composition for preventing or treating lymphoma, comprising poceltinib as an active ingredient, wherein the pharmaceutical composition is administered in combination with an immunomodulatory drug.

[0067] The pharmaceutical composition according to the present invention can be prepared according to conventional methods in the pharmaceutical field. The pharmaceutical composition can be combined with an appropriate pharmaceutically acceptable carrier depending on the dosage form, and, if necessary, can be prepared by further including excipients, diluents, dispersants, emulsifiers, buffers, stabilizers, binders, disintegrants, solvents, etc. The appropriate carriers, etc., which do not inhibit the activity and properties of the pharmaceutical composition according to the present invention, can be selected differently depending on the dosage form and dosage form.

[0068] The pharmaceutical composition of the present invention may additionally include suitable carriers, excipients, or diluents commonly used in the manufacture of pharmaceutical compositions. Compositions containing pharmaceutically acceptable carriers may be administered in various oral or parenteral dosage forms. When formulated, the compositions may be prepared using commonly used diluents or excipients, such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants.

[0069] Solid dosage forms for oral administration may include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms may be prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid dosage forms for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included.

[0070] Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0071] In addition, the pharmaceutical composition of the present invention may have any one dosage form selected from the group consisting of, but not limited to, tablets, pills, powders, granules, capsules, suspensions, liquid solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories.

[0072] Each component of the pharmaceutical composition according to the present invention may be included in the pharmaceutical composition in a pharmaceutically effective amount.

[0073] The above pharmaceutical composition can be administered in a pharmaceutically effective amount.

[0074] The above pharmaceutically effective amount may vary depending on the intended use, the patient's age, sex, weight, and health condition, the type and severity of the disease, the activity of the drug, the sensitivity to the drug, the method of administration, the time of administration, the route of administration, and the excretion rate, the duration of treatment, the drugs used in combination or concurrently, and other factors well known in the medical field. For example, although not fixed, it may generally be administered in the range of 1 to 1000 mg / kg, for example, 10 to 400 mg / kg once or several times daily. The above dosage does not limit the scope of the present invention in any way.

[0075] In another aspect of the present invention, the immunomodulatory drug may be at least one selected from the group consisting of lenalidomide, thalidomide, pomalidomide, imiquimod, rituximab, mezigdomide, iberdomide, golcadomide, ICP-490, GLB-002, GT919, GT929, HP-001, BTX-1188, and cemsidomidide, but is not limited thereto. Specifically, the immunomodulatory drug may be lenalidomide or cemsidomidide.

[0076] In another aspect of the present invention, a pharmaceutical composition for preventing or treating lymphoma comprising Poseltinib and an immunomodulatory drug as active ingredients may further comprise, as an active ingredient, at least one selected from the group consisting of Bruton's Tyrosine Kinase Inhibitors (BTKi), immunosuppressants, targeted anticancer agents, immune checkpoint inhibitors, angiogenesis inhibitors, cytokines and cytokine modulators, cancer vaccines, small molecule compound inhibitors, epigenetic modification modulators, antibodies, anti-CD19 antibodies, anti-CD20 antibodies, anti-CD20 / anti-CD3 bispecific antibodies, antibody-drug conjugates, and chimeric antigen receptor T (CAR-T) cell therapy agents, but is not limited thereto.

[0077] The pharmaceutical composition according to the present invention may further comprise a surfactant. Surfactants include, but are not limited to, lipids such as phospholipids, phosphatidylcholine, lecithin, cardiolipin, fatty acids, phosphatidylethanolamine, phosphatides, tyloxapol, polyethylene glycol, PEG 400, PEG 1500, PEG 2000, poloxamer 407, poloxamer 188, polysorbate 80, polysorbate 20, sorbitan lithium, sorbitan stearate, sorbitan palmitate, or mixtures thereof.

[0078] The pharmaceutical composition according to the present invention may further comprise a stabilizer or gelling agent. Such stabilizers or gelling agents include, but are not limited to, propylene glycol monopalmitostearate, glyceryl monostearate, glyceryl dibehenate, glyceryl distearate, hydrogenated fats, polyvinylpyrrolidone, polyethylene, glycerol, polyoxyethylene stearate, sorbitan fatty acid esters, cholesterol, macrogol-20-glycerol monostearate, poloxamer 124, isopropyl myristate, isopropyl palmitate, colloidal silica, hydrophobic colloidal silica, magnesium stearate, zinc stearate, aluminum stearate, lanolin alcohol, organoclay, petrolatum or polyoxyl 6 stearate.

[0079] The pharmaceutical composition according to the present invention may further comprise a polymer-based carrier. Polymer-based carriers include, but are not limited to, acrylic polymers such as polyacrylic and polymethacrylic acid derivatives, such as cellulose, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), carboxymethyl cellulose (CMC), methylcellulose (MC), hydroxyethylcellulose (HEC), amylase, amylopectin, dextran, polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), HEMA, carbopol, derivatives thereof, or mixtures thereof.

[0080] The pharmaceutical composition according to the present invention may further comprise an organic cosolvent. Organic cosolvents include, but are not limited to, ethylene glycol, propylene glycol, N-methyl pyrrolidone, 2-pyrrolidone, 3-pyrrolidinol, 1,4-butanediol, dimethylglycol monomethyl ether, diethylene glycol monomethyl ether, solketal, glycerol, polyethylene glycol, or polypropylene glycol.

[0081] The pharmaceutical composition according to the present invention may additionally comprise a pH-active ingredient. Suitable pH-active ingredients, such as buffers or pH-adjusting agents, include, but are not limited to, disodium phosphate, monosodium phosphate, boric acid, sodium borate, sodium citrate, hydrochloric acid, or sodium hydroxide.

[0082] The pharmaceutical composition according to the present invention may additionally comprise an osmotically active ingredient. Examples of osmotically active ingredients include, but are not limited to, sodium chloride, mannitol, or glycerol.

[0083] The pharmaceutical composition according to the present invention may further comprise a preservative. Preservatives include, but are not limited to, benzalkonium chloride, alkyldimethylbenzylammonium chloride, cetrimide, cetylpyridinium chloride, benzododecinium bromide, benzethonium chloride, thiomersal, chlorobutanol, benzyl alcohol, phenoxethanol, phenylethyl alcohol, sorbic acid, methyl, propyl paraben, chlorhexidine digluconate, EDTA, or mixtures thereof.

[0084] In addition, the pharmaceutical composition according to the present invention may include a sustained-release form, such as, but not limited to, a gel formulation, a liposome formulation, a lipid microemulsion formulation, a microsphere formulation, a nanosphere formulation, or an implant formulation, to provide the active compound continuously.

[0085] In another aspect of the present invention, a pharmaceutical composition for preventing or treating lymphoma, comprising the above-described posseltinib as an active ingredient, which is administered in combination with an immunomodulatory drug, may be administered in combination with any one or more selected from the group consisting of Bruton's Tyrosine Kinase Inhibitors (BTKi), immunosuppressants, targeted anticancer agents, immune checkpoint inhibitors, angiogenesis inhibitors, cytokines and cytokine modulators, cancer vaccines, small molecule compound inhibitors, epigenetic modification modulators, antibodies, anti-CD19 antibodies, anti-CD20 antibodies, anti-CD20 / anti-CD3 bispecific antibodies, antibody-drug conjugates, and chimeric antigen receptor T (CAR-T) cell therapy agents, but is not limited thereto.

[0086] In another aspect of the present invention, the BTKi may be at least one selected from the group consisting of Ibrutinib, Acalabrutinib, Zanubrutinib, Tirabrutinib, Orelabrutinib, Spebrutinib, Evobrutinib, Vecabrutinib, Pirtobrutinib, and Fenebrutinib, but is not limited thereto.

[0087] The pharmaceutical composition may be appropriately administered to a subject according to a conventional method, administration route, and dosage used in the art, depending on the purpose or need. Examples of administration routes include oral, parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal administration, and parenteral injection includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In addition, an appropriate dosage and frequency of administration may be selected according to a method known in the art, and the actual amount of the pharmaceutical composition of the present invention to be administered and the frequency of administration may be appropriately determined by various factors, such as the type of symptom to be treated, administration route, sex, health condition, diet, age and weight of the subject, and severity of the disease.

[0088] The pharmaceutical composition may be administered to any animal capable of developing lymphoma, particularly diffuse large B-cell lymphoma (DLBCL) and primary central nervous system lymphoma (PCNSL), and the animal may include, for example, humans and primates, as well as livestock such as cows, pigs, horses, and dogs. In some embodiments, the animal may be an animal other than a human.

[0089] The above pharmaceutical composition can be administered via an appropriate route of administration depending on the formulation, and can be administered via various routes, either oral or parenteral, as long as it can reach the target tissue. The method of administration is not particularly limited, and can be administered by conventional methods such as oral, rectal, intravenous, intramuscular, transdermal, or respiratory inhalation.

[0090] The pharmaceutically effective amount of each active ingredient used in combination may vary depending on the specific compound or pharmaceutical composition used, the mode of administration, the condition to be treated, the severity of the condition to be treated, the species of warm-blooded animal, body weight, sex, diet, and age. Therefore, the dosage regimen using the pharmaceutical composition of the present invention is selected based on various factors, including the route of administration and the patient's renal and hepatic function. A surgeon, clinician, or veterinarian skilled in the art can readily determine and prescribe the effective amount of the drug required to prevent, respond to, or arrest the progression of the condition. Optimal precision in achieving drug concentrations within the range of efficacy without toxicity requires a regimen based on the dynamics of drug availability targeting the site. This includes considering the distribution, equilibrium, and elimination of the drug. Therefore, the dosage regimen, i.e., the dosage level and frequency of administration of any individual component of the present invention, can be adjusted to provide the optimal therapeutic response.

[0091] In another aspect of the present invention, the immunosuppressant, targeted anticancer agent, anti-CD19 antibody, anti-CD20 antibody, anti-CD20 / anti-CD3 bispecific antibody, antibody drug conjugate or chimeric antigen receptor T (CAR-T) cell therapy agent is selected from the group consisting of cyclophosphamide, doxorubicin, etoposide, gemcitabine, liposomal doxorubicin, prednisone, procarbazine, vincristine, selinexor, dexamethasone, tafasitamab, ofatumumab, obinutuzumab, rituximab, It may be any one selected from the group consisting of, but is not limited to, mosunetuzumab, epcoritamab, glofitamab, polatuzumab vedotin, brentuximab vedotin, loncastuximab tesirine, tisagenlecleucel, axicabtagene ciloleucel, and lisocabtagene maraleucel.

[0092] In another aspect of the present invention, the lymphoma may be non-Hodgkin lymphoma or Hodgkin lymphoma.

[0093] Specifically, the non-Hodgkin's lymphoma may be any one selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), primary central nervous system lymphoma (PCNSL), follicular lymphoma, primary mediastinal B-cell lymphoma, mantle cell lymphoma, post-transplant lymphoproliferative disorder, marginal zone lymphoma, Waldenstrom's macroglobulinemia, and Burkitt lymphoma, but is not limited thereto.

[0094] More specifically, the non-Hodgkin's lymphoma may be diffuse large B-cell lymphoma (DLBCL) or primary central nervous system lymphoma (PCNSL).

[0095] In another aspect of the present invention, a composition comprising posseltinib and an immunomodulatory drug, such as lenalidomide, may have a synergistic effect on the prevention or treatment of lymphoma.

[0096] In particular, the mixing ratio of posseltinib and an immunomodulatory drug, such as lenalidomide, in the composition of the present invention can be determined to exhibit maximum therapeutic efficacy and minimum side effects in actual clinical application by comprehensively considering the degree of synergistic effect according to drug interaction shown in in vitro experimental results evaluated at the cancer cell level, and the actual tumor growth inhibition efficacy and pharmacokinetic characteristics observed in an in vivo experimental environment using an animal model. This optimal ratio may vary depending on the specific cancer type, type of animal model, route of administration, etc.

[0097] Specifically, the process of determining the ratio of posseltinib and lenalidomide included in the composition can be determined comprehensively by considering not only the direct pharmacological action of the two drugs on cancer cells under in vitro conditions, particularly the ratio that maximizes the synergistic effect, but also the complex drug behavior characteristics in the in vivo environment. In vitro experiments can be determined by considering the interaction and effect of the drugs themselves on specific cancer cells, and in vivo experiments can be determined by considering various biological factors such as drug delivery through blood circulation, distribution to various tissues, drug-metabolizing enzyme activity in the liver and other organs, and potential interactions with immune cells. Therefore, the ratio that maximizes therapeutic efficacy in actual clinical settings can be determined by comprehensively considering in vitro and in vivo data.

[0098] Therefore, in the actual clinical trial stage, the administration ratio of posseltinib and lenalidomide can be finely adjusted by comprehensively considering the individual characteristics of the patient (e.g., age, gender, underlying disease), type and stage of cancer, and presence or absence of other concomitant treatments.

[0099] Poseltinib and an immunomodulatory drug, such as lenalidomide, may be administered at a ratio of 10:1 to 1:100. Specifically, poseltinib and an immunomodulatory drug, such as lenalidomide or semcidomide, may be administered at a ratio of 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, or 1:1. Alternatively, the above-mentioned posseltinib and immunomodulatory drugs such as lenalidomide or semcidomide may be administered at a concentration ratio of 1:100, 1:90, 1:80, 1:70, 1:60, 1:50, 1:40, 1:30, 1:20, 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, or 1:1.

[0100] Preferably, the above-mentioned posseltinib and immunomodulatory drugs such as lenalidomide or semcidomide may be administered at a concentration ratio of 10:1 to 1:1, 9:1 to 1:1, 8:1 to 1:1, 7:1 to 1:1, 6:1 to 1:1, 5:1 to 1:1, 4:1 to 1:1, 3:1 to 1:1, 2:1 to 1:1, or 1:1 to 1:1. Alternatively, the concentration ratio may be 8:1 to 1:1, 8:1 to 2:1, 8:1 to 3:1, 8:1 to 4:1, or 8:1 to 5:1.

[0101] In another aspect of the present invention, the effective ingredients may be administered simultaneously or sequentially.

[0102] In another aspect of the present invention, the composition may have a synergistic effect on the prevention or treatment of lymphoma.

[0103] Methods for prevention or treatment, and uses

[0104] In one aspect of the present invention, the present invention relates to a method for preventing or treating lymphoma, comprising administering to a subject a therapeutically effective amount of a pharmaceutical composition comprising poceltinib and an immunomodulatory drug as active ingredients.

[0105] In one aspect of the present invention, the present invention relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of posseltinib and an immunomodulatory drug as active ingredients for preparing a medicament for the prevention or treatment of lymphoma.

[0106] In another aspect of the present invention, the present invention relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of poceltinib as an active ingredient for preparing a medicament for the prevention or treatment of lymphoma, the pharmaceutical composition being administered in combination with an immunomodulatory drug.

[0107] In one aspect of the present invention, the present invention relates to a use of a pharmaceutical composition comprising a therapeutically effective amount of poceltinib and an immunomodulatory drug as active ingredients for the prevention or treatment of lymphoma.

[0108] In another aspect of the present invention, the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of poceltinib as an active ingredient, the pharmaceutical composition being administered in combination with an immunomodulatory drug, for the prevention or treatment of lymphoma.

[0109] Among the terms or elements mentioned in the above methods and uses, the composition of the pharmaceutical composition is understood to be the same as mentioned in the description of the pharmaceutical composition and combination above.

[0110] Hereinafter, to aid understanding of the present invention, examples will be given in detail. However, the following examples are intended only to illustrate the content of the present invention and are not intended to limit the scope of the present invention to the following examples.

[0111]

[0112] Example 1: Single drug treatment of poseltiniib, lenalidomide, and ibrutinib

[0113] We aimed to determine the single treatment effects of posseltinib, lenalidomide, and ibrutinib on non-Hodgkin's lymphoma cell lines OCI-LY-19 (DSMZ), SU-DHL-5 (DSMZ), and TMD8 (Seoul National University).

[0114] First, the above cells to be cultured are placed in a cell culture medium at 5x10 4 cells / ml concentration and dispensed 100 μl per well into a 96-well plate (Corning, 3598). Poseltinib (Hanmi Pharmaceutical Co., Ltd.), ibrutinib (Selleckchem Co., Ltd., S2680), and lenalidomide (Selleckchem Co., Ltd., S1029) were diluted 2-fold to the final target concentrations of 0.00001 uM, 0.0001 uM, 0.001 uM, 0.1 uM, 1 uM, and 10 uM, respectively, in cell culture medium (RPMI1640, 84%; FBS, 15%; Penicillin-Streptomycin, 1%) and dispensed 100 μl per well into the 96-well plate. At this time, the total volume per well was 200 μl. After that, it was cultured in a CO2 incubator at 37℃ for 72 hours.

[0115] Afterwards, 20 ul of WST-8 (BIOMAX, QM1000), corresponding to 10% of the total volume per well, was treated, and the reaction was performed in a 37℃ CO2 incubator for 2 to 4 hours. After that, the cell viability was confirmed by measuring the absorbance at 450 nm using a Microplate reader (Molecular Devices, SpectraMax190).

[0116] As a result, when poseltiniib, lenalidomide, and ibrutinib were treated alone at a concentration of 10 uM in the OCI-LY-19 cell line, cell viability decreased to approximately 70%, and when poseltiniib, lenalidomide, and ibrutinib were treated alone at a concentration of 10 uM in the SU-DHL-5 cell line, cell viability also decreased to approximately 80%.

[0117] Meanwhile, in the TMD8 cell line, when treated with posseltinib at a concentration of 0.1 uM, cell viability decreased to approximately 60%, when treated with a concentration of 1 uM, it decreased to approximately 30%, and when treated with a concentration of 10 uM, it decreased to less than 30%. In the case of lenalidomide, when treated with a concentration of 10 uM, cell viability decreased to less than 30%, and in the case of ibrutinib, when treated with a concentration of 0.001 uM, it decreased to approximately 70%, and when treated with a concentration of 0.01 uM or higher, it decreased to less than approximately 30% (Fig. 1). That is, ibrutinib showed a high cell killing rate at a low concentration when treated as a single agent.

[0118] Example 2: Effect of combination treatment of poseltiniib-lenalidomide and ibrutinib-lenalidomide (Drug combination treatment)

[0119] The aim was to determine the effect of combined drug administration on non-Hodgkin's lymphoma cell lines OCI-LY-19, SU-DHL-5, and TMD8.

[0120] Cells (OCI-LY-19, SU-DHL-5, and TMD8) and drugs (poseltiniib, lenalidomide, and ibrutinib) were prepared in the same manner as in Example 1. Two drug combinations (poseltiniib and lenalidomide; ibrutinib and lenalidomide) for which the synergistic effect of combination treatment was to be confirmed were diluted 4-fold to the target concentrations of 0.0001 uM, 0.001 uM, 0.1 uM, 1 uM, and 10 uM, respectively, and dispensed 50 μl per well into the 96-well plate. At this time, the total volume per well became 200 μl. Thereafter, they were cultured in a CO2 incubator at 37°C for 72 hours.

[0121] Afterwards, 20 ul of WST-8 (BIOMAX, QM1000), corresponding to 10% of the total volume per well, was treated, and the reaction was performed in a 37℃ CO2 incubator for 2 to 4 hours. After that, the cell viability was confirmed by measuring the absorbance at 450 nm using a Microplate reader (Molecular Devices, SpectraMax190).

[0122] Example 2-1: TMD8 cell line (1:1 ratio)

[0123] This study aimed to investigate the combination treatment effect of poseltiniib-lenalidomide and ibrutinib-lenalidomide at a 1:1 ratio in the TMD8 cell line. In particular, the synergism index, which indicates the synergistic effect of the two drugs, was investigated. The synergism index was calculated by using the concentrations and survival rates of the two co-administered substances as a ZIP (Zero Interaction Potency) score, and R (version 4.4.2) and the synergyfinder (version 3.14.0) package were used. The formula for calculating the ZIP score is as follows. To analyze the combination treatment effect, a drug concentration-response curve was generated from the single drug treatment group, and the difference between the predicted combination effect and the actual observed combination effect was quantified as a ZIP score to evaluate the synergistic effect.

[0124]

[0125] x1, x2: concentration of each experimental drug; λ1, λ2: shape parameters representing the sigmoid or slope of the concentration-response curve of each experimental drug; m1, m2: median of the maximum and minimum effects of each experimental drug.

[0126] To achieve a 1:1 ratio of poseltiniib-lenalidomide concentration, poseltiniib and lenalidomide were combined in a 1:1 ratio at concentrations ranging from 0 uM to 10 uM, respectively, and treated to the TMD8 cell line. Ibrutinib-lenalidomide was also treated in the same manner to determine cell viability.

[0127] Additionally, based on the results of cell viability according to the concentration of the above drugs, the synergistic effect of combined treatment between the two drugs was confirmed.

[0128] As in Example 1 above, when the effect of single drug treatment was confirmed, ibrutinib showed the best effect, and when ibrutinib-lenalidomide was combined, a lower cell viability was observed even at a lower concentration than when poseltiniib-lenalidomide was combined (Fig. 2).

[0129] However, in the case of the synergy index, the average for poseltiniib-lenalidomide was 1.90, the maximum was 6.31, and the average for ibrutinib-lenalidomide was -1.19, the maximum was 4.65, showing that the synergy effect of poseltiniib-lenalidomide was higher (Figure 3).

[0130] Example 2-2: TMD8 cell line (1:50, 1:1000 ratio)

[0131] We aimed to determine the combined treatment effects of poseltiniib-lenalidomide at a 1:50 ratio and ibrutinib-lenalidomide at a 1:1000 ratio in TMD8 cell lines. In particular, we aimed to determine the synergy index, which indicates the synergistic effect of the two drugs.

[0132] In the case of ibrutinib, the highest cell death rate was observed in the single treatment in Example 1. When measuring the synergy index of the two drugs, it is necessary to conduct the experiment by setting the concentration to an appropriate level of cell death rate that does not kill all cells when each drug is treated alone. Therefore, in the case of ibrutinib-lenalidomide, the experiment was conducted at a lower concentration ratio of 1:1000 than that of posseltinib-lenalidomide, which was set at a concentration ratio of 1:50.

[0133] First, to achieve a posseltinib-lenalidomide concentration ratio of 1:50, posseltinib was prepared at a concentration of 0 uM to 0.4 uM, and lenalidomide was prepared at a concentration of 0 uM to 20 uM, and treated to the TMD8 cell line.

[0134] Additionally, to achieve an ibrutinib-lenalidomide ratio of 1:1000, ibrutinib was prepared at a concentration of 0 uM to 0.02 uM, and lenalidomide was prepared at a concentration of 0 uM to 20 uM, and the TMD8 cell line was treated to determine cell viability.

[0135] Additionally, based on the results of cell viability at different drug concentrations, the synergistic effect of the combination treatment between the two drugs was confirmed. The synergy index was calculated as a ZIP (Zero Interaction Potency) score using the concentration and viability of the two substances administered together, and R (version 4.4.2) and the synergyfinder (version 3.14.0) package were used. The formula for calculating the ZIP score is as follows. To analyze the combination treatment effect, a drug concentration-response curve was generated from the single drug treatment group, and the difference between the predicted combination effect and the actually observed combination effect was quantified as a ZIP score to evaluate the synergistic effect.

[0136]

[0137] x1, x2: concentration of each experimental drug; λ1, λ2: shape parameters representing the sigmoid or slope of the concentration-response curve of each experimental drug; m1, m2: median of the maximum and minimum effects of each experimental drug.

[0138] At this time, similar levels of cell viability were achieved when posseltinib-lenalidomide was co-treated at a concentration of 1:50 and when ibrutinib-lenalidomide was co-treated at a concentration of 1:1000 (Fig. 4).

[0139] Meanwhile, the synergy index was confirmed to be 19.49 for posseltinib-lenalidomide and 13.52 for ibrutinib-lenalidomide, indicating that the combination treatment of posseltinib-lenalidomide showed a much better synergy effect (Figure 5).

[0140] Example 2-3: OCI-LY19 cell line (1:1 ratio)

[0141] We aimed to investigate the combined treatment effects of poseltiniib-lenalidomide and ibrutinib-lenalidomide at a 1:1 ratio in the OCI-LY19 cell line. In particular, we aimed to determine the synergy index, which indicates the synergistic effect of the two drugs.

[0142] To achieve a 1:1 ratio of poseltiniib-lenalidomide concentration, poseltiniib and lenalidomide were combined in a 1:1 ratio at concentrations ranging from 0 uM to 10 uM, respectively, and treated to the OCI-LY19 cell line. Ibrutinib-lenalidomide was also treated in the same manner to determine cell viability.

[0143] Additionally, based on the results of cell viability at different drug concentrations, the synergistic effect of the combination treatment between the two drugs was confirmed. The synergy index was calculated as a ZIP (Zero Interaction Potency) score using the concentration and viability of the two substances administered together, and R (version 4.4.2) and the synergyfinder (version 3.14.0) package were used. The formula for calculating the ZIP score is as follows. To analyze the combination treatment effect, a drug concentration-response curve was generated from the single drug treatment group, and the difference between the predicted combination effect and the actually observed combination effect was quantified as a ZIP score to evaluate the synergistic effect.

[0144]

[0145] x1, x2: concentration of each experimental drug; λ1, λ2: shape parameters representing the sigmoid or slope of the concentration-response curve of each experimental drug; m1, m2: median of the maximum and minimum effects of each experimental drug.

[0146] At this time, no significant difference in cell viability was confirmed when combined treatment with poseltiniib-lenalidomide and ibrutinib-lenalidomide (Fig. 6).

[0147] In terms of synergy indices, no significant difference was observed between poseltiniib-lenalidomide and ibrutinib-lenalidomide (Fig. 7). However, considering that ibrutinib achieved a higher cell death rate even at a lower concentration than poseltiniib when used alone, as confirmed in Example 1, the fact that both poseltiniib-lenalidomide and ibrutinib-lenalidomide had similar synergy indices when ibrutinib and poseltiniib were co-treated with lenalidomide at the same concentration and ratio, respectively, indicates the advantageous effect of poseltiniib-lenalidomide.

[0148] Example 2-4: SU-DHL-5 cell line (1:1 ratio)

[0149] We aimed to investigate the combined treatment effects of poseltiniib-lenalidomide and ibrutinib-lenalidomide at a 1:1 ratio in the SU-DHL-5 cell line. In particular, we aimed to determine the synergy index, which indicates the synergistic effect of the two drugs.

[0150] To achieve a 1:1 ratio of poseltiniib-lenalidomide concentration, poseltiniib and lenalidomide were combined in a 1:1 ratio at concentrations ranging from 0 uM to 10 uM, respectively, and treated to the SU-DHL-5 cell line. Ibrutinib-lenalidomide was also treated in the same manner to determine cell viability.

[0151] Additionally, based on the results of cell viability at different drug concentrations, the synergistic effect of the combination treatment between the two drugs was confirmed. The synergy index was calculated as a ZIP (Zero Interaction Potency) score using the concentration and viability of the two substances administered together, and R (version 4.4.2) and the synergyfinder (version 3.14.0) package were used. The formula for calculating the ZIP score is as follows. To analyze the combination treatment effect, a drug concentration-response curve was generated from the single drug treatment group, and the difference between the predicted combination effect and the actually observed combination effect was quantified as a ZIP score to evaluate the synergistic effect.

[0152]

[0153] x1, x2: concentration of each experimental drug; λ1, λ2: shape parameters representing the sigmoid or slope of the concentration-response curve of each experimental drug; m1, m2: median of the maximum and minimum effects of each experimental drug.

[0154] At this time, no significant difference in cell viability was confirmed when combined treatment with poseltiniib-lenalidomide and ibrutinib-lenalidomide (Fig. 8).

[0155] In terms of synergy indices, no significant difference was observed between poseltiniib-lenalidomide and ibrutinib-lenalidomide (Fig. 9). However, considering that ibrutinib achieved a higher cell death rate at a lower concentration than poseltiniib when used alone, as confirmed in Example 1, the fact that poseltiniib-lenalidomide and ibrutinib-lenalidomide had similar synergy indices when ibrutinib and poseltiniib were co-treated with lenalidomide at the same concentration and ratio, respectively, indicates the advantageous effect of poseltiniib-lenalidomide.

[0156] Example 3: Effect of combination treatment of posseltinib and lenalidomide using a xenograft model (drug combination treatment)

[0157] We created a xenograft animal model by transplanting the MYD88 / CD79B mutant TMD8 (Tokyo medical and dental university) tumor cell line into CB-17 SCID mice (Saeronbio) and examined the antitumor effect of combined administration of posseltinib and lenalidomide.

[0158] MYD88 / CD79B mutant TMD8 cell line was cultured in RPMI 1640 (Gibco, Cat. No. 22400-089) medium containing 10% NBCS (Gibco, Cat. No. 26010074) at 37°C and 5% CO2. The cultured cells were obtained and resuspended in 100 μL of Hank's Balanced Salt solution (HBSS, Gibco, Cat. No. 14170112). The suspended cells were plated on BD Matrigel. TM After mixing in a 2:1 ratio with the matrix, 0.15 mL (1 x 10) was injected subcutaneously in the flank of 28 8-week-old male CB-17 SCID mice. 7 Xenograft animals were created by injecting each cell.

[0159] Vehicle, poseltiniib alone, lenalidomide alone, and poseltiniib-lenalidomide combination were orally administered to the produced xenograft animals for 29 days as shown in Table 1.

[0160] [Table 1] Composition of TMD8 Xenograft animal groups and administration method for each group

[0161]

[0162] During the administration period, the body weight of animals was measured two to three times a week. In addition, the tumor size was measured three times a week using a digital caliper (MITUTOYO CD-15CPX, Japan). The tumor volume was calculated using the formula for calculating the volume of an ellipsoid, V (mm 3 ) = L × S 2 / 2(V, tumor volume; L, long diameter; S, short diameter) was used to calculate the tumor inhibition effect. The tumor inhibition effect was compared by calculating the inhibition rate, which was calculated by subtracting the tumor growth rate of the drug administration group from 100% and the tumor growth rate of the vehicle group (%). The tumor growth rate (relative tumor volume) was calculated by dividing the tumor volume on the day of measurement by the tumor volume on the first day. The maximum inhibition rate means the highest inhibition rate during the observation period.

[0163] When body weight loss was observed in each animal, no body weight loss was observed when 50 mg / kg and 100 mg / kg of poseltinibe were administered, and a maximum body weight loss of 0.5% was observed on day 2 when 30 mg / kg of lenalidomide was administered. When 50 mg / kg of poseltinibe and 30 mg / kg of lenalidomide were co-administered, a maximum body weight loss of 1.4% was observed on day 12, and when 100 mg / kg of poseltinibe and 30 mg / kg of lenalidomide were co-administered, a maximum body weight loss of 0.8% was observed on day 9, indicating that no significant body weight loss was observed when poseltinibe and lenalidomide were co-administered (Fig. 10).

[0164] In terms of tumor suppression effect, when poseltiniib 50 mg / kg was administered as a single agent, the maximum suppression effect was -6.1% (day 5), when poseltiniib 100 mg / kg was administered, it was 68.5% (day 21), and when lenalidomide 30 mg / kg was administered, it was 24.9% (day 21). On the other hand, in combination administration, when poseltiniib 50 mg / kg and lenalidomide 30 mg / kg were co-administered, the maximum suppression effect was 51.9% (day 28), and when poseltiniib 100 mg / kg and lenalidomide 30 mg / kg were co-administered, the maximum suppression effect was 81.4% (day 29), indicating that combination administration was more effective (Fig. 11).

[0165] Example 4: Effect of combination treatment of posseltinib and semcidomide (drug combination treatment)

[0166] The purpose of this study was to investigate the effect of combined treatment of posseltinib and semcidomide on SU-DHL-5. Cultured SU-DHL-5 cells were seeded at a density of 1x10 using cell medium (RPMI1640, 10% FBS, 1% sodium pyruvate, 1% HEPES, 1% Penicillin-Streptomycin). 5 After diluting to a concentration of 10 cells / mL, 100 μL was dispensed per well into a 96-well plate (Corning, 3598) (1x10 4 cells / well). Afterwards, they were cultured in a 37℃ CO2 incubator for 24 hours (recovery step).

[0167] After completion of the culture, poseltinibrix (Hanmi Pharmaceutical Co., Ltd.) and cemsidomide (Medchemexpress Co., Ltd., HY-144841) were prepared at 4 times the target concentration using cell medium and 50 μL was added to each well. When treating with one drug, 50 μL of cell medium was added. At this time, the total volume per well was 200 μL, and the target concentrations of each drug were set to 0, 4, 5, 6, 7, and 8 μM for poseltinibrix and 0, 0.00001, 0.0001, 0.001, 0.01, and 0.1 nM for cemsidomide. After that, the cells were cultured in a CO2 incubator at 37°C for 72 hours.

[0168] Afterwards, 20 μL of WST-8 (BIOMAX, QM1000) reagent, corresponding to 10% of the total volume per well, was treated, and the reaction was performed in a 37°C CO2 incubator for 2 to 3 hours. After that, the cell proliferation rate was confirmed by measuring the absorbance at 450 nm using a Microplate reader (Molecular Devices, SpectraMax, ABP00433).

[0169] The synergy index was calculated using the ZIP (Zero Interaction Potency) score using the concentrations and survival rates of the two co-administered substances, and the R (version 4.4.2) and synergyfinder (version 3.14.0) packages were used. The formula for calculating the ZIP score is as follows. To analyze the combination treatment effect, a drug concentration-response curve was generated from the single drug treatment group, and the difference between the predicted combination effect and the actually observed combination effect was quantified as the ZIP score to evaluate the synergy effect.

[0170]

[0171] x1, x2: concentration of each experimental drug; λ1, λ2: shape parameters representing the sigmoid or slope of the concentration-response curve of each experimental drug; m1, m2: median of the maximum and minimum effects of each experimental drug.

[0172] As a result, when the two substances were combined in the SU-DHL-5 cell line, a high cell survival inhibition effect was observed, and the average synergy index was confirmed to be 23.89, indicating a very excellent synergy effect (Fig. 12).

[0173] Example 5: Effect of poseltinibet on the treatment of primary central nervous system lymphoma (PCNSL)

[0174] In Example 2, the synergistic effect of poseltiniib-lenalidomide was confirmed, and the therapeutic effect of poseltiniib on primary central nervous system lymphoma (PCNSL) was examined.

[0175] To treat PCNSL with BTK inhibitors, high brain penetration is required because they must cross the blood-brain barrier (BBB) ​​to reach the central nervous system.

[0176] Example 5-1: Confirmation of brain penetration of posseltinib

[0177] After orally administering 10 mg / kg of poseltiniib to male Sprague-Dawley rats (Orient Bio, Korea), the concentrations of poseltiniib in plasma and brain tissue were measured by LC / MS (Liquid Chromatography Tandem Mass Spectrometry) method.

[0178] At this time, the peak concentration (C max ) was measured as 4.5 ng / ml, and the brain penetration rate was calculated as 16.8% (Table 2). This result is higher than the brain penetration rate of other BTK inhibitors using the same rats.

[0179] [Table 2]

[0180]

[0181] Example 5-2: Confirmation of therapeutic effect on PCNSL

[0182] A phase 2 clinical trial was conducted on the treatment of PCNSL with combination therapy with posseltinib (CRIS registration number: KCT0008105). The trial consisted of phases 1 and 2. In the phase 1 induction trial, the efficacy of the combination therapy with rituximab, lenalidomide, and posseltinib was confirmed. In the phase 2 consolidation trial, the efficacy of the combination therapy with lenalidomide 15 mg (QD) and posseltinib 40 mg (BID) was confirmed. As a result of administering the drugs as combination therapy to the seven recruited patients, three patients responded, resulting in an overall response rate of 42.9%. Of these, two patients showed complete response, confirming an excellent antitumor effect with a complete response rate of 28.6%.

Claims

1. Poseltinib; and Containing an immunomodulatory drug as an active ingredient, A pharmaceutical composition for the prevention or treatment of lymphoma.

2. A pharmaceutical composition for the prevention or treatment of lymphoma, comprising posseltinib as an active ingredient, wherein the pharmaceutical composition is administered in combination with an immunomodulatory drug.

3. In paragraph 1 or 2, A pharmaceutical composition, wherein the immunomodulatory drug is at least one selected from the group consisting of Lenalidomide, Thalidomide, Pomalidomide, Imiquimod, Rituximab, Mezigdomide, Iberdomide, Golcadomide, ICP-490, GLB-002, GT919, GT929, HP-001, BTX-1188, and Cemsidomide.

4. In paragraph 3, A pharmaceutical composition wherein the above immunomodulatory drug is lenalidomide or semsidomide.

5. In paragraph 1, The pharmaceutical composition may further include, as an active ingredient, at least one selected from the group consisting of Bruton's Tyrosine Kinase Inhibitors (BTKi), immunosuppressants, targeted anticancer agents, immune checkpoint inhibitors, angiogenesis inhibitors, cytokines and cytokine modulators, cancer vaccines, small molecule compound inhibitors, epigenetic modification modulators, antibodies, anti-CD19 antibodies, anti-CD20 antibodies, anti-CD20 / anti-CD3 bispecific antibodies, antibody-drug conjugates, and chimeric antigen receptor T (CAR-T) cell therapy agents.

6. In paragraph 2, The pharmaceutical composition may be administered in combination with at least one selected from the group consisting of Bruton's Tyrosine Kinase Inhibitors (BTKi), immunosuppressants, targeted anticancer agents, immune checkpoint inhibitors, angiogenesis inhibitors, cytokines and cytokine modulators, cancer vaccines, small molecule compound inhibitors, epigenetic modification modulators, antibodies, anti-CD19 antibodies, anti-CD20 antibodies, anti-CD20 / anti-CD3 bispecific antibodies, antibody-drug conjugates, and chimeric antigen receptor T (CAR-T) cell therapy agents.

7. A pharmaceutical composition according to claim 5 or 6, wherein the BTKi is at least one selected from the group consisting of Ibrutinib, Acalabrutinib, Zanubrutinib, Tirabrutinib, Orelabrutinib, Spebrutinib, Evobrutinib, Vecabrutinib, Pirtobrutinib, and Fenebrutinib.

8. In paragraph 1 or 2, A pharmaceutical composition, wherein the above lymphoma is non-Hodgkin lymphoma or Hodgkin lymphoma.

9. In paragraph 8, A pharmaceutical composition, wherein the non-Hodgkin's lymphoma is any one selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), primary central nervous system lymphoma (PCNSL), follicular lymphoma, primary mediastinal B-cell lymphoma, mantle cell lymphoma, post-transplant lymphoproliferative disorder, marginal zone lymphoma, Waldenstrom's macroglobulinemia, and Burkitt lymphoma.

10. In paragraph 8, A pharmaceutical composition, wherein the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma (DLBCL) or primary central nervous system lymphoma (PCNSL).

11. In paragraph 4, A pharmaceutical composition wherein the above posseltinib and lenalidomide can be administered at a concentration ratio of 10:1 to 1:

100.

12. In paragraph 11, A pharmaceutical composition wherein the above posseltinib and lenalidomide can be administered at a concentration ratio of 10:1 to 1:

1.

13. A pharmaceutical composition according to claim 1 or 2, wherein the effective ingredients can be administered simultaneously or sequentially.

14. A pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition has a synergistic effect on the prevention or treatment of lymphoma.

15. A method for preventing or treating lymphoma, comprising administering to a subject a therapeutically effective amount of a pharmaceutical composition comprising posseltinib and an immunomodulatory drug as active ingredients.

16. Use of a pharmaceutical composition comprising a therapeutically effective amount of posseltinib and an immunomodulatory drug as active ingredients for the prevention or treatment of lymphoma.

17. Use of a pharmaceutical composition comprising a therapeutically effective amount of posseltinib and an immunomodulatory drug as active ingredients for manufacturing a medicament for preventing or treating lymphoma.

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