Combinations for immunomodulation in cancer treatment

Abstract

To provide a pharmaceutical composition for use in a method for treating a cancer or refractory cancer and / or inhibiting cancer metastasis, recurrence or progression.SOLUTION: A pharmaceutical composition includes CX-4945 or a pharmaceutically acceptable salt or ester thereof, the method including the step for administering a combination of (i) the CX-4945 or pharmaceutically acceptable salt or ester thereof and (ii) an anti-CTLA-4 antibody to subjects, the CX-4945 having the following structure.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] This invention relates to the field of cancer treatment. In particular, this invention provides a method for treating cancer using a combination of chemotherapeutic agents, CK2 inhibitors, and immune checkpoint inhibitors. [Background technology]

[0002] Protein kinase CK2 is upregulated in many human cancers and has therefore been considered a target for cancer therapy (Chua et al., Pharmaceuticals 2017, 10, 18). CK2 inhibitors have been shown to inhibit cell proliferation and may be used to treat neoplastic disorders such as breast cancer, prostate cancer, pancreatic cancer, lung cancer, hematopoietic cancer, colorectal cancer, skin cancer, and ovarian cancer (Chua et al., U.S. Patent No. 9,062,043 B2). However, CK2 inhibitors alone may only have a moderate anticancer effect against certain types of cancer.

[0003] Several series of CK2 inhibitors have been studied for their activity in inhibiting cell growth, both alone and in combination with other antiproliferative agents (U.S. Patent No. 9,062,043 B2, U.S. Patent No. 7,956,064 B2; Publications WO 2010080170 A1 and WO 2011011199 A1). WO 2010080170 A1 discloses that CK2 inhibitors may be combined with anticancer agents that inhibit cell growth, such as alkylating agents, antimetabolites, vinca alkaloids, taxanes, topoisomerase inhibitors, antitumor antibiotics, and tyrosine kinase inhibitors, to treat or improve neoplastic disorders. Furthermore, WO 2010008170 A1 also discloses that CK2 inhibitors may be combined with immunosuppressive macrolides to treat neoplastic disorders.

[0004] Certain CK2 inhibitors have also been shown to be combined with inhibitors of molecules essential to the cell growth pathway, such as AKT inhibitors, HDAC inhibitors, HSP90 inhibitors, mTOR inhibitors, PBK / niTGR inhibitors, PDK inhibitors, and antibodies targeting tumor / cancer antigens, to treat or improve neoplastic disorders and / or inflammatory, autoimmune, or infectious disorders (WO 2011011199 A1). [Overview of the Initiative] [Problems that the invention aims to solve]

[0005] However, there is a need to identify novel compounds that can enhance the immune response against tumor cells. [Means for solving the problem]

[0006] In one embodiment, the Disclosure provides a method for treating cancer or refractory cancer in a subject and / or inhibiting cancer metastasis, recurrence or progression, or increasing the likelihood of survival for an appropriate period of time in a subject diagnosed with cancer, the method comprising the step of administering to the subject a combination of an anticancer agent, a CK2 inhibitor and optionally an immune checkpoint inhibitor.

[0007] In one embodiment, the anticancer agent is (i) an antihormone agent that acts to regulate or inhibit the hormonal action on the tumor, such as tamoxifen, raloxifen, droloxifen, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY117018, onapristone, and toremifene citrate. (ii) Aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production in the adrenal gland, such as 4(5)-imidazole, aminoglutethimide, megestrol acetate, exemestane, formestanie, fadrozol, borozole, letrozole and anastrozole; (iii) Antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; and troxacitabine (1,3- (iv) dioxolane nucleoside cytosine analogs; (v) protein kinase inhibitors, e.g. MEK inhibitors, e.g. cobimetinib; (v) lipid kinase inhibitors, e.g. taselicib; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signaling pathways involved in abnormal cell proliferation, e.g., PKC-alpha, Raf, and H-Ras, e.g., oblimersen; (vii) ribozymes, e.g., VEGF expression inhibitors and HER2 expression inhibitors; (viii) vaccines, e.g., gene therapy vaccines, topoisomerase 1 inhibitors; (ix) angiogenesis inhibitors, e.g., bevacizumab; and any pharmaceutically acceptable salts, acids, and derivatives of any of the above. In another embodiment, the anticancer agent is a therapeutic antibody, e.g., alemtuzumab, bevacizumab, cetuximab, panitumumab, rituximab, pertuzumab, trastuzumab, trastuzumab, emtansine, and tocitumomab.

[0008] In a further embodiment, the anti-cancer agent is a chemotherapeutic agent including, but not limited to, cisplatin, gemcitabine, carboplatin, methotrexate, vincristine, adriamycin, bleomycin, hydroxyurea, salinosporamide A, bortezomib, PS-519, omuralid, cyclophosphamide, ifosfamide, 5-fluorouracil, vinblastine, docetaxel, doxorubicin, and paclitaxel.

[0009] In some embodiments of the present invention, the CK2 inhibitor is a compound having the structure of Formula I or a pharmaceutically acceptable salt or ester thereof

Chemical formula

[0010] In a particular preferred embodiment of the present invention, the CK2 inhibitor is CX-4945.

[0011] In one embodiment, the immunotherapy agent is an immune checkpoint inhibitor. In some embodiments of the present invention, the immune checkpoint inhibitor is a PD-1 antagonist or a CTLA-4 antagonist. In some embodiments, the immune checkpoint inhibitor is a CTLA-4, PD-1, or PD-1 antibody. In some embodiments, the PD-1 or CTLA-4 inhibitor includes, but is not limited to, humanized antibodies that block human PD-1, such as lambrolizumab (anti-PD-1 Ab, trademark Keytruda) or pizilizumab (anti-PD-1 Ab), nivolumab (anti-PD-1 Ab, trademark Opdivo), tisilimucob (anti-CTLA-4 Ab), and ipilimumab (anti-CTLA-4 Ab).

[0012] In one embodiment, the administration induces immunological memory against the cancer. In another embodiment, the administration increases the number of T cells and activated T cells in the tumor microenvironment.

[0013] In one embodiment, the combination of anticancer agents and CK2 inhibitors may be administered simultaneously, sequentially, intermittently, or periodically.

[0014] A treatment protocol appropriate for treating a subject with cancer or for inhibiting cancer metastasis, recurrence, or progression in a subject includes, for example, administering at least one administration cycle.

[0015] In one embodiment, the method comprises at least one administration cycle, the cycle having a duration of 8 weeks or less. In a further embodiment, the cycle has a duration of 4 weeks.

[0016] In one embodiment, a treatment cycle involves administering to a subject one or more first treatments (e.g., induction therapy, e.g., a combination of an anticancer agent and a CK2 inhibitor) in a dose and regimen sufficient to achieve a response (partial or complete response).

[0017] In one embodiment, the administration cycle includes administering a CK2 inhibitor daily and an anticancer drug once a week. In a further administration, the administration cycle further includes administering an immune checkpoint inhibitor twice a week.

[0018] In one embodiment, the chemotherapeutic agent, CK2 inhibitor, and optionally an immune checkpoint inhibitor are present in synergistically effective amounts.

[0019] In one embodiment, a chemotherapeutic agent, a CK2 inhibitor, and optionally an immune checkpoint inhibitor are administered in doses effective in increasing the immune response against cancer in the subject.

[0020] In some embodiments, the CK2 inhibitor is administered daily at a dose ranging from approximately 25 mg (approximately 25 mg / kg) to approximately 2,000 mg / kg of the subject's body weight. In some embodiments, the daily dose is in the range of approximately 50 mg / kg to approximately 200 mg / kg. In some embodiments, the CK2 inhibitor is administered twice daily.

[0021] In some embodiments, the anticancer drug is administered in a dose ranging from approximately 1 mg (approximately 1 mg / kg) to approximately 20 mg / kg per kg of the subject's body weight. In some embodiments, the dose is in the range of approximately 5 mg / kg to approximately 10 mg / kg.

[0022] In some embodiments, immune checkpoint inhibitors are administered in doses ranging from approximately 1 mg (approximately 1 mg / kg) to approximately 20 mg / kg of the subject's body weight. In some embodiments, the dose ranges from approximately 8 mg / kg to approximately 15 mg / kg.

[0023] In some embodiments, cancers include, but are not limited to, skin cancers (e.g., melanoma and basal cell carcinoma), glioblastoma, liver cancers (e.g., hepatocellular carcinoma), colorectal cancer, glioblastoma, gastric cancer, colorectal cancer, esophageal cancer, lung cancers (e.g., non-small cell lung cancer (NSCLC) and small cell lung cancer), pancreatic cancer, renal cell carcinoma, benign prostatic hyperplasia, prostate cancer, ovarian cancer, melanoma, breast cancer, chronic lymphocytic leukemia (CLL), Merkel cell carcinoma, non-Hodgkin lymphoma, acute myeloid leukemia (AML), gallbladder cancer, bile duct cancer, bladder cancer, and uterine cancer.

[0024] According to the present invention, the subject may also be further administered by CAR-T therapy.

[0025] In some embodiments, administration of a therapeutically effective dose of a concomitant therapeutic agent reduces the expression of IL-6 mRNA or protein in the subject.

[0026] In some embodiments, administration of a therapeutically effective dose of a combination therapy increases the target's immune response to the tumor. In some embodiments, the increased immune response may be measured as a greater number or activity of T cells in the tumor microenvironment of the treated target, fewer MDSCs systemically or in the tumor microenvironment, or fewer TAMs in the tumor microenvironment. [Brief explanation of the drawing]

[0027] [Figure 1] This figure shows the average tumor volume in an ovarian tumor model. [Figure 2] This figure shows the average tumor volume in a colorectal cancer model. [Figure 3] This figure shows the 14-day antitumor growth activity of CX-4945 after cisplatin treatment in the immunocompetent syngeneic mouse model 4T1. [Figure 4] This figure shows tumor-infiltrating CD4 and CD8 T cells in the syngeneic mouse model 4T1 after treatment with CX-4945 and / or cisplatin. [Figure 5] This figure shows the Kaplan-Meier survival curves for subjects treated with a vehicle, CX-4945 at 37.5 mg / kg, anti-CTLA4 at 10 mg / kg, and a combination of CX-4945 and anti-CTLA-4. [Figure 6] This figure shows the tumor growth curves (average tumor volume over time) for naive mice and mice whose cancer was cured. [Figure 7] This figure shows the results of the change in average body weight in tumor-bearing mice. [Figure 8] This figure shows the Kaplan-Meier survival curves for subjects in a Phase 1 clinical trial, where patients received at least one cycle of CX-4945 in combination with cisplatin and gemcitabine without dose changes or reductions. [Modes for carrying out the invention]

[0028] It should be understood that the technical terms used herein are for the sole purpose of describing specific embodiments and are not intended to limit them.

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art in which the present invention pertains. Generally, the nomenclature and experimental methods used herein, as described below, are well known and commonly used in the art.

[0030] As used herein, the terms “a,” “an,” and “the,” as well as similar references used in this context, may be interpreted as encompassing both singular and plural forms.

[0031] Unless otherwise indicated, all figures used in this specification and the claims, representing quantities of components, reaction conditions, etc., should be understood in all cases to be modified by the term “approximately.” Therefore, unless otherwise indicated, the numerical parameters shown in this specification and the appended claims are approximations that may vary depending on the desired properties to be obtained by this application. Generally, as used herein, the term “approximately,” when referring to measurable values ​​such as weight, time, or dose, means encompassing a variation from a particular quantity of, in one example, ±15% or ±10%, in another ±5%, in yet another ±1%, and in yet another ±0.1%, such variation itself being appropriate for carrying out the disclosed method.

[0032] As used herein, the term “therapeutic dose” means an amount sufficient to perform such treatment for a disease, disorder, or condition when administered to an animal for the treatment of a disease.

[0033] As used herein, the terms “treatment,” “to treat,” and “to treat” mean reversing, reducing, delaying the onset of a disease or disorder or one or more of its symptoms, or inhibiting the progression of a disease or disorder or one or more of its symptoms, as described herein. In some embodiments, the treatment may be administered after the onset of one or more symptoms. In other embodiments, the treatment may be administered in the absence of symptoms.

[0034] As used herein, “pharmaceutically acceptable” means suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, etc., and effective for intended use within the bounds of sound medical judgment, commensurate with a reasonable benefit-risk ratio.

[0035] "Salt" includes derivatives of activators, which are modified by producing acid or base addition salts thereof. Preferably, the salt is a pharmaceutically acceptable salt. Such salts include, but are not limited to, pharmaceutically acceptable acid addition salts, pharmaceutically acceptable base addition salts, pharmaceutically acceptable metal salts, ammonium and alkylated ammonium salts. Acid addition salts include salts of inorganic and organic acids. Typical examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, and nitric acid. Typical examples of suitable organic acids include formic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, propionic acid, benzoic acid, cinnamic acid, citric acid, fumaric acid, glycolic acid, lactic acid, maleic acid, malic acid, malonic acid, mandelic acid, oxalic acid, picric acid, pyruvic acid, salicylic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, tartaric acid, ascorbic acid, pamoic acid, bismethylenesalicylic acid, ethanedisulfonic acid, gluconic acid, citraconic acid, aspartic acid, stearic acid, palmitic acid, EDTA, glycolic acid, p-aminobenzoic acid, glutamic acid, benzenesulfonic acid, p-toluenesulfonic acid, sulfates, nitrates, phosphates, perchlorates, borates, acetates, benzoates, hydroxynaphthoates, glycerophosphates, and ketoglutarates. Base addition salts include, but are not limited to, ethylenediamine, N-methylglucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris-(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabiethylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids such as lysine and arginine dicyclohexylamine. Examples of metal salts include lithium salts, sodium salts, potassium salts, and magnesium salts.Examples of ammonium salts and alkylated ammonium salts include ammonium salt, methylammonium salt, dimethylammonium salt, trimethylammonium salt, ethylammonium salt, hydroxyethylammonium salt, diethylammonium salt, butylammonium salt, and tetramethylammonium salt. Examples of organic bases include lysine, arginine, guanidine, diethanolamine, and choline. Standard methods for preparing pharmaceutically acceptable salts and their formulations are well known in the art and are disclosed in various references, including, for example, "Remington: The Science and Practice of Pharmacy," edited by A. Gennaro, 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.

[0036] As used herein, the term “synergistically effective” means that the combined effect of two or more therapeutic agents used in combination is greater than their additive effect when used individually.

[0037] As used herein, the terms “subject,” “individual,” or “patient” are interchangeable herein and refer to vertebrates, preferably mammals, and more preferably humans.

[0038] As used herein, the term “immunotherapy agent” refers to chemical and biological products that modulate a person’s immune response to produce a desired therapeutic effect.

[0039] As used herein, the terms “immunosuppressive cancer” or “immunosuppressive cancer” refer to cancer associated with immunosuppressive properties in the subject, either systemically or within the tumor microenvironment. These immunosuppressive properties include any one of the following: high regulated T cells, high myelo-derived suppressor cells (MDSCs), high tumor-associated macrophages (TAMs), low presence or activity of CD4 or CD8 T cells, and low activity of antigen-presenting cells (APCs).

[0040] The tumor microenvironment is the cellular environment in which a tumor resides, including surrounding blood vessels, hematopoietic precursors, immune cells, fibroblasts, extracellular matrix, and signaling molecules. Immune cells include lymphocytes and other hematopoietic cells involved in immune function (e.g., antigen-presenting cells, macrophages, neutrophils, NK cells, monocytes, and myeloid suppressor cells). Signaling molecules include cytokines, chemokines, growth factors, and other soluble proteins that function in influencing cell activity and composition.

[0041] CK2 inhibitors have been shown to act synergistically with immune checkpoint modulators (ICMs), such as anti-PD1 antibodies or anti-cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) antibodies. Both PD-1 and CTLA4 are known to suppress costimulatory signaling, which is essential for T cell activation. However, such CK2 inhibitors do not possess effective tumor growth inhibitory (TGI) properties when used alone (WO 2017 / 070137 A1). Furthermore, a range of CK2 inhibitors, namely BMS-211, BMS-699, and BMS-595, have been shown to reduce polymorphonuclear MDSCs (PMN-MDSCs) and tumor-associated macrophages (TAMs), which are negative regulators of tumor-specific immune responses, and this is likely to contribute to their synergistic effects with immune checkpoint inhibitors (Hashimoto et al., Cancer Res 78: 5644-5655).

[0042] The fact that simply antagonizing suppressive signaling molecules in T cells, such as PD-1 or CTLA-4, is insufficient to control tumor growth via the tumor-specific cytotoxic activity of T cells suggests that additional immune components within the tumor microenvironment may contribute to immunosuppressive mechanisms that help tumors evade T cell-mediated killing.

[0043] Accordingly, the present disclosure provides a method for treating cancer or refractory cancer in a subject and / or inhibiting cancer metastasis, recurrence or progression, or increasing the likelihood of survival over an appropriate period of time in a subject diagnosed with cancer, the method comprising the step of administering to the subject a combination of an anticancer agent, a CK2 inhibitor and optionally an immune checkpoint inhibitor.

[0044] The anticancer agents used in this disclosure may also be chemotherapeutic agents. Chemotherapeutic agents include conventional chemotherapeutic reagents, such as alkylating agents, antimetabolites, plant alkaloids, antibiotics and other compounds, such as cisplatinum, CDDP, methotrexate, vincristine, adriamycin, bleomycin and hydroxyurea. Chemotherapeutic agents also include proteasome inhibitors, such as salinosporamide (e.g., salinosporamide A), bortezomib, PS-519 and omralid. The most commonly used types of anticancer drugs include DNA alkylating agents (e.g., cyclophosphamide, ifosfamide), antimetabolites (e.g., methotrexate, folate antagonists, and 5-fluorouracil, pyrimidine antagonists), microtubule disruptors (e.g., vincristine, vinblastine, paclitaxel), DNA intercalators (e.g., doxorubicin, daunomycin, cisplatin), and hormone therapies (e.g., tamoxifen, flutamide). Other platinum-coordinated complexes evaluated in clinical trials include carboplatin, tetraplatin, olmiplatin, iproplatin, and oxaliplatin (see Kelland, Crit. Rev. Oncol. Hematol, 15: 191-219 (1993)).

[0045] Alkylating agents include (a) alkylating-like platinum-based chemotherapeutic agents, such as cisplatin, carboplatin, nedaplatin, oxaliplatin, satoraplatin and (SP-4-3)-(cis)-aminedichloro-[2-methylpyridine]platinum(II); (b) alkyl sulfonates, such as busulfan; (c) ethyleneimine and methylmelamine derivatives, such as altoretamine and thiotepa; (d) nitrogen mustard, for example (e) chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechloretamine, trophosamide, prednimustine, melphalan and uramustine; (f) nitrosourea, such as carmustine, lomustine, fotemustine, nimustine, ranimustine and streptozocin; (g) triazenes and imidazotetrazines, such as dacarbazine, procarbazine, temozolamide and temozolomide.

[0046] According to certain embodiments of the present invention, the chemotherapeutic agent may be selected from the group consisting of cisplatin, methotrexate, vincristine, adriamycin, bleomycin, hydroxyurea, salinosporamide A, bortezomib, PS-519, omralid, cyclophosphamide, ifosfamide, methotrexate, 5-fluorouracil, vinblastine, and paclitaxel.

[0047] CK2 inhibitors can offer anti-cancer and anti-inflammatory potential. CK2 inhibitors are generally classified into three categories: (1) inhibitors that target regulatory subunits of CK2 (e.g., genetically selected peptide aptamers); (2) inhibitors of the catalytic activity of CK2 (e.g., quinobene, TBB, DMAT, IQA); and (3) CK2 holoenzyme disruptors, which are often molecules that bind to the CK2 subunit interface and inhibit the high-affinity interactions of those subunits. Each class of CK2 inhibitor may be any type of molecule, such as small molecules, functional nucleic acids, antibodies, or peptide mimes.

[0048] CK2 catalytic subunits possess constitutive activity. However, in eukaryotic cells, the CK2β subunit is not only a central component of the tetrameric CK2 complex but is also involved in the recruitment of CK2 substrates. Therefore, the dynamic interactions of CK2 subunits observed in living cells may play a crucial role in the CK2 signaling pathway. Drugs that specifically target this interaction are less likely to have side effects than drugs that act as general inhibitors of CK2 catalytic activity.

[0049] CK2 inhibitors consist of a wide variety of chemical substances, including flavonoids (e.g., apigenin), hydroxyanthraquinone / xanthenone derivatives (e.g., emodin), hydroxycoumarin derivatives (e.g., DBC), tetrabromotriazole / imidazole derivatives (e.g., DRB, TBB, DMAT, TBCA, TBBz), and indoloquinazoline derivatives (e.g., IQA).

[0050] In certain embodiments, the CK2 inhibitor is a compound represented by formula I as described herein. According to certain embodiments of the present invention, the CK2 inhibitor may be selected from the group consisting of 4,5,6,7-tetrabromobenzotriazole (TBB), quinalizarin, hematein, tetrabromocinnamic acid (TBCA), CIGB-300, CX-4945, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), apigenin, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT), emodin, 5-oxo-5,6-dihydro-indro(1,2-a)quinazoline-7-yl]acetic acid (IQA), CX-4945, and 6,7-dichloro-1,4-dihydro-8-hydroxy-4-[(4-methylphenylamino)methylene]dibenzo[b,d]furan-3(2H)-one (TF).

[0051] In particular, the CK2 inhibitor is CX-4945 (silmitasertib), which has the following structure:

[0052] [ka]

[0053] Immunotherapy agents are drugs that can modulate the physiological levels of one or more cytokines within the tumor microenvironment of cancer. In some embodiments, immunotherapy agents induce local production of at least one endogenous cytokine, e.g., TNF-α or IFN-γ, which has a cytotoxic effect on tumor cells. In some embodiments, immunotherapy agents inhibit the production of endogenous cytokines, e.g., IL-10, TGFβ, or VEGF, which interfere with T cell recognition and the destruction of cancer cells. In other embodiments, immunotherapy agents are drugs that can induce tumor cell production of one or more chemokines that attract immune cells, e.g., dendritic cells, effector T cells (e.g., CD8+ lymphocytes), and natural killer (NK) cells to tumor cells. In some embodiments, chemokines include, but are not limited to, CCL19, CCL20, CCL21, CX3CL1, CXCL9, and CXCL10.

[0054] In other embodiments, immunotherapeutic agents are drugs that induce immune checkpoint blockade, such as PD-1 blockade and CTLA-4 blockade. Immune checkpoint inhibitors are known to provide some antitumor activity in humans, although this partial antitumor activity is observed in only a small fraction of the treated subjects. Immune checkpoint inhibitors include antagonists of inhibitory receptors that inhibit the PD-1 or CTLA-4 pathway, such as anti-PD-1, anti-PD-L1, or anti-CTLA-4 antibodies or inhibitors. Examples of PD-1 or PD-L1 inhibitors include, but are not limited to, humanized antibodies that block human PD-1, such as lambrolizumab (anti-PD-1 Ab, trade name Keytruda) or pizilizumab (anti-PD-1 Ab), Bavencio (anti-PD-L1 Ab, avelumab), Imfinzi (anti-PD-L1 Ab, durvalumab) and Tecentriq (anti-PD-L1 Ab, atezolizumab), as well as fully human antibodies, such as nivolumab (anti-PD-1 Ab, trade name Opdivo). Other PD-1 inhibitors may include, but are not limited to, other PD-1 inhibitors currently under investigation and / or development for use in presentation and therapy of soluble PD-1 ligands containing PD-L2 Fc fusion proteins, also known as B7-DC-Ig or AMP-244. Furthermore, immune checkpoint inhibitors may include, but are not limited to, humanized or fully human antibodies that block PD-L1, such as durvalumab and MIH1, as well as other PD-L1 inhibitors currently under investigation.

[0055] The active ingredients described herein may be pharmaceutical compositions prepared by commonly used methods using excipients commonly used in this field, i.e., pharmaceutical excipients, pharmaceutical carriers, etc.

[0056] Tablets, powders, granules, etc., are used as solid compositions for oral administration. In such solid compositions, one, two, or more types of active ingredients are mixed with at least one inactive excipient. The composition may also contain inactive additives, such as lubricants, disintegrants, stabilizers, and solvents, in commonly used manner.

[0057] Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solution preparations, suspensions, syrups, or elixirs, and commonly used inert diluents, such as purified water or ethanol. In addition to the inert diluent, the liquid composition may also contain adjuvants, such as solubilizers, wetting agents and suspension agents, sweeteners, flavorings, fragrances, or preservatives.

[0058] Injections for parenteral administration contain sterile water or non-aqueous preparations, suspensions, or emulsions. Aqueous solvents include, for example, distilled water for injection or physiological saline. Non-aqueous solvents include, for example, alcohols, such as ethanol. Such compositions may further contain isotonic agents, preservatives, wetting agents, emulsifiers, dispersants, stabilizers, or solvents. These are sterilized, for example, by filtration with a bacterial-retaining filter, mixing with a bactericide, or irradiation. Furthermore, these may also be used in a manner in which a sterile solid composition is prepared and dissolved or suspended in sterile water or a sterile solvent for injection before use.

[0059] Transmucosal preparations, such as nasal preparations, are used in solid, liquid, or semi-solid form and can be prepared according to methods known in the relevant field. For example, known excipients, pH adjusters, preservatives, surfactants, lubricants, stabilizers, and thickeners may be added as appropriate. For administration, an appropriate device can be used for inhalation or inhalation.

[0060] Co-administration may include the simultaneous administration of an anticancer agent, a CK2 inhibitor, and optionally an immune checkpoint inhibitor, or separate administration of the therapeutic agents, in the same or different dosage forms. For example, the anticancer agent, the CK2 inhibitor, and optionally an immune checkpoint inhibitor may be administered simultaneously. Alternatively, the anticancer agent may be administered in combination with the CK2 inhibitor and optionally an immune checkpoint inhibitor, and the anticancer agents that can be administered in combination with the CK2 inhibitor and optionally an immune checkpoint inhibitor may be formulated for separate administration and administered simultaneously or sequentially.

[0061] The following examples provide a more detailed description of certain aspects and embodiments of the present disclosure, but they are merely illustrative and should not be considered to limit the scope of the claims. The present invention includes, for example, the following embodiments: [Embodiment 1] A method for treating cancer or refractory cancer in a subject and / or inhibiting cancer metastasis, recurrence or progression, or increasing the chances of survival over an appropriate period of time in a subject diagnosed with cancer, comprising the step of administering a combination of an anticancer agent, a CK2 inhibitor and optionally an immune checkpoint inhibitor to the subject. [Embodiment 2] The method according to Embodiment 1, wherein administration increases tumor-specific T cells in the tumor microenvironment of the target. [Embodiment 3] The method according to Embodiment 1, wherein the combination of an anticancer drug and a CK2 inhibitor can be administered simultaneously, sequentially, intermittently, or periodically. [Embodiment 4] The method according to Embodiment 1, comprising at least one administration cycle, wherein the cycle has a duration of 8 weeks or less. [Embodiment 5] The method according to Embodiment 1, wherein the combination provides immunological memory against the aforementioned cancer. [Embodiment 6] The method according to Embodiment 1, wherein the anticancer agent, CK2 inhibitor, and optionally an immune checkpoint inhibitor are present in synergistically effective amounts. [Embodiment 7] The method according to Embodiment 4, wherein the cycle has a period of 4 weeks. [Embodiment 8] The method according to Embodiment 4, wherein the administration cycle comprises administering to the subject one or more first treatments (e.g., induction therapy, e.g., a combination of a chemotherapy agent and a CK2 inhibitor) in an amount and regimen sufficient to achieve a response (partial or complete response), and then administering to the subject a certain amount of an immune checkpoint inhibitor. [Embodiment 9] The method according to Embodiment 4, wherein the administration cycle includes administering a CK2 inhibitor daily and a chemotherapeutic agent once a week. [Embodiment 10] The method according to Embodiment 9, further comprising administering an immune checkpoint inhibitor twice a week as part of the administration cycle. [Embodiment 11] The method according to Embodiment 4, wherein the CK2 inhibitor is administered at a daily dose in the range of approximately 25 mg (approximately 25 mg / kg) to approximately 2000 mg / kg per kg of the subject's body weight. [Embodiment 12] The method according to Embodiment 4, wherein the daily dose is in the range of approximately 50 mg / kg to approximately 200 mg / kg, and in some embodiments, the CK2 inhibitor is administered twice a day. [Embodiment 13] The method according to Embodiment 4, wherein the anticancer drug is administered in a dose ranging from approximately 1 mg (approximately 1 mg / kg) to approximately 20 mg / kg per kg of the subject's body weight. [Embodiment 14] The method according to Embodiment 4, wherein the dose of the anticancer drug is in the range of approximately 5 mg / kg to approximately 10 mg / kg. [Embodiment 15] The method according to Embodiment 4, wherein the immune checkpoint inhibitor is administered in a dose ranging from approximately 1 mg (approximately 1 mg / kg) to approximately 20 mg / kg per kg of the subject's body weight. [Embodiment 16] The method according to Embodiment 4, wherein the dose of the immune checkpoint inhibitor is in the range of approximately 8 mg / kg to approximately 15 mg / kg. [Embodiment 17] The method according to Embodiment 1, wherein the cancer is glioblastoma, skin cancer (e.g., melanoma and basal cell carcinoma), liver cancer (e.g., hepatocellular carcinoma), colorectal cancer, glioblastoma, gastric cancer, colorectal cancer, esophageal cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC) and small cell lung cancer), pancreatic cancer, renal cell carcinoma, benign prostatic hyperplasia, prostate cancer, ovarian cancer, melanoma, breast cancer, chronic lymphocytic leukemia (CLL), Merkel cell carcinoma, non-Hodgkin lymphoma, acute myeloid leukemia (AML), gallbladder cancer, bile duct cancer, bladder cancer, or uterine cancer. [Embodiment 18] The method according to Embodiment 1, wherein the anticancer agent is cisplatin, gemcitabine, carboplatin, methotrexate, vincristine, adriamycin, bleomycin, hydroxyurea, salinosporamide A, bortezomib, PS-519, omralid, cyclophosphamide, ifosfamide, docetaxel, doxorubicin, 5-fluorouracil, vinblastine, or paclitaxel. [Embodiment 19] The CK2 inhibitor is a compound having the structure of formula I or a pharmaceutically acceptable salt or ester thereof. [ka] (In the formula, each Z 1 、Z 2 、Z 3 and Z 4 is N or CR 3 And, Z 5 、Z 6 、Z 7 and Z 8 Each of these is N or CR 6 And, Z 1 ~Z 4 Of these, 0, 1, or 2 are N, and Z 5 ~Z 8 Of these, 0, 1, or 2 are N, and Z 1 ~Z 4 and Z 5 ~Z 8 At least one of them is a nitrogen atom, Each R 3 and each R 6 Independently, H is a C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl or C6-C12 heteroarylalkyl, or Each R 6 These are independently: Halo, OR, NR 2 NROR, NRNR 2 , SR, SOR, SO 2 R, SO 2 NR 2 NRSO 2 R, NRCONR 2 ,NRCOOR,NRCOR,CN,OC(O)R,COR,NO 2 , or polar substituents selected from carboxylic acids, carboxylates, esters, carboxamides, tetrazoles, or

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[0062] [Examples]

[0063] [Example 1] CX-4945 exhibits in vivo synergy with the DNA damaging agent gemcitabine. In the A2780 ovarian tumor model, a significant increase in time to endpoint was demonstrated with the combination of gemcitabine (Gem) (120 mg / kg) and CX-4945 (25 mg / kg or 100 mg / kg) after only four doses, compared to administration of each drug individually (Figure 1). The Colo-205 colorectal cancer model was resistant to each drug alone (Figure 2). However, the combination of gemcitabine and CX-4945 significantly reduced tumor growth, suggesting that this combination may be effective in indications where gemcitabine alone is not approved.

[0064] [Example 2] Enhancement of antitumor effects by drug combinations in animal cancer models Table 1 shows that the drug combinations described in the present invention yield synergistic antineoplastic effects in vivo. The use of 60 mg / kg gemcitabine by IV injection and 100 mg / kg CX-4945 by oral administration to A2780 ovarian cancer xenografts was well-tolerated and significantly enhanced antitumor activity compared to gemcitabine alone. Time to endpoint (TTE) analysis showed a significant delay in time to endpoint for gemcitabine, CX-4945, and combinations of both drugs.

[0065] [Table 1]

[0066] [Example 3] CX-4945 significantly increases T cells in tumors and synergistically inhibits tumor growth in vivo when used in conjunction with chemotherapy agents. The efficacy of tumor growth inhibition and T cell activation studies were conducted in 4T1, an immunocompetent syngeneic mouse model known to be resistant to immune checkpoint inhibitors, such as anti-PD1, anti-PD-L1, and anti-CTLA4. Resistance to these antagonists, which modulate T cell costimulatory signaling, indicates a highly immunosuppressive tumor microenvironment.

[0067] This series of studies was conducted in 6 - 8 week - old Balb / C mice. The breast cancer cell line 4T1 was subcutaneously injected into the right flank of the mice using 3×10 5 cells in 0.1 ml of PBS. The mice were randomized for treatment when the average tumor size reached approximately 80 - 120 mm 3 . They were administered CX - 4945 and / or cisplatin at the doses and schedules described in Table 1. CX - 4945 was formulated in a 25 mM Na2HPO4 solution (pH = 9.2) and administered by forced oral gavage. Cisplatin was dissolved in a saline buffer and administered by intraperitoneal injection. Tumors were measured two - dimensionally (width × length) using an electronic caliper, and the tumor volume was calculated using the formula V=(L×W×W) / 2, where V is the tumor volume, L is the tumor length (the longest dimension of the tumor), and W is the tumor width (the longest dimension of the tumor perpendicular to L). The mice were monitored twice a week and euthanized for flow cytometry analysis when the tumor size reached approximately 800 - 1000 mm 3 .

[0068]

Table 2

[0069] Antitumor activity was measured as the percentage tumor growth inhibition (%TGI) at the time of euthanasia. TGI was calculated as %TGI = 100×(1 - T / C). T and C were the mean tumor volumes of the treatment and control groups, respectively, on a given day. The 14 - day antitumor activity is shown in Table 2, and the tumor growth curve is shown in Figure 3. In summary, CX - 4945 shows a synergistic effect with cisplatin, a platinum - based DNA - damaging agent, in inhibiting tumor growth.

[0070] The composition of immune cells was analyzed when the tumor size was 800 - 1000 mm 3On the day when [the specified condition] was reached, it was analyzed using multi-color flow cytometry (FACS). The tumor was treated with lysis medium (Miltenyi, CAT#130 - 096 - 730) and resuspended in FACS buffer. T cell infiltration was measured by examining the number of T cells in the tumor sample. CD4 T cells, CD8 T cells, activated CD4 T cells, and activated CD8 T cells were identified by cell surface markers as CD45 + CD3 + CD4 + , CD45 + CD3 + CD8 + , CD45 + CD3 + CD4 + CD69 + and CD45 + CD3 + CD8 + CD69 + respectively. The absolute number of cells was measured by the normalized number of cells using counting beads (eBiosciences CAT# 01 - 1234 - 42) and the weight of each tumor sample. The absolute number of tumor - infiltrating T cells was calculated by the formula: absolute number (cells / mg) = [(number of cells × eBead volume) / (number of eBeads × tumor weight)] × eBead concentration.

[0071] Treatment of immunocompetent mice with CX - 4945 does not reduce the number of MDSCs in the tumor microenvironment either systemically or locally. Furthermore, treatment of immunocompetent mice with CX - 4945 and cisplatin unexpectedly increases both CD4 and CD8 T cells in the tumor microenvironment. In particular, the combination of CX - 4945 and cisplatin showed a synergistic effect in increasing the number of T cells and activated T cells in the tumor microenvironment (Figure 4).

[0072] The tumor growth inhibition by the combination of CX - 4945 and cisplatin was associated with cytotoxic T cell activity in the tumor microenvironment. Depleting CD8 T cells by administering an anti - CD8 antibody along with the treatment resulted in the disappearance of the antitumor growth activity.

[0073] [Example 4] The combination of CX-4945 and anti-CTL4 significantly increases the chances of survival in patients with cancer. The ability of CX-4945 to enhance T cell activity for effective tumor killing was also tested in 4T1, the same syngeneic model that has demonstrated refractory status to anti-PD1 or anti-CTLA4 treatment. This series of studies was conducted in 6-8 week old Balb / C mice. 4T1 breast cancer cells were placed in 0.1 ml PBS at a rate of 3 × 10⁶ cells. 5 The cells were subcutaneously injected into the right flank of mice. The mice had an average tumor size of approximately 80-120 mm. 3 When the mice reached a certain stage, they were randomized for treatment. The mice were then treated with 37.5 mg / kg of CX-4945 administered orally and / or 10 mg / kg of anti-CTLA4 antibody administered intraperitoneally. Kaplan-Meier survival curves were generated and log-rank tests were performed. 3000m 3 Mice with growing tumors were considered to have reached the tumor growth endpoint. The combination of CX-4945 and anti-CTLA4 significantly improved the overall survival of tumor-bearing mice (Figure 5), and 28.5% of the group treated with such a combination achieved complete remission.

[0074] Surviving mice (i.e., mice cured of cancer) that reached the efficacy phase (4T-1) (complete tumor regression) were re-challenged with the same cancer cell line, 4T-1, to confirm whether the mice could develop immunity to the same cancer cell line. After tumor cell inoculation, mice (naive mice and cancer-cured mice) were checked daily for morbidity and mortality. During routine monitoring, animals were checked for any effects on tumor growth and treatment behavior, such as exercise, food and water consumption, weight gain / loss (weight was measured twice a week), and any other abnormalities. Mortality and observed clinical signs were recorded for each individual animal. Tumor growth curves (mean tumor volume over time) for different groups are shown in Figure 6. The results of mean weight changes in tumor-bearing mice are shown in Figure 7. The results showed that mice cured of cancer from initial treatment with CX4945 and anti-mouse CTLA-4 antibody could not develop 4T-1 tumors after re-challenge, suggesting acquired immunity after drug treatment. In conclusion, surviving mice treated with a combination of CX-4945 and an anti-CTLA4 antibody showed superior activity in inhibiting tumor growth when re-challenged with 4T1 tumor cells, demonstrating successful induction of immunological memory.

[0075] In summary, activated T cell activity and immunological memory within the tumor microenvironment induced by the combined CX-4945 regimen contribute to the long-term survival of subjects with tumors.

[0076] [Example 5] Comparison of clinical trial results The ABC-02 trial collected data from 208 patients, 138 of whom had received first-line chemotherapy with gemcitabine and cisplatin (Juan Valle et al., N. Engl. J. Med. 362:1273-1281 (2010)). See Table 3 below.

[0077] [Table 3]

[0078] For comparison, Table 4 below shows the results of a Phase I / II study of CX-4945 in combination with gemcitabine and cisplatin as a frontline treatment for patients with cholangiocarcinoma.

[0079] [Table 4]

[0080] [Example 6] Treatment with CX-4945 in combination with cisplatin and gemcitabine increases overall survival in patients with cholangiocarcinoma. In a Phase I, open-label, multicenter, multi-dose, dose-escalation study of CX-4945 in combination with gemcitabine and cisplatin in patients with cholangiocarcinoma, 50 patients with unresectable cholangiocarcinoma were administered CX-4945 in combination with gemcitabine and cisplatin in 21-day cycles. CX-4945 was administered intravenously with cisplatin 25 mg / m² on days 1 and 8. 2 and gemcitabine 1,000 mg / m² 2 The drug was administered orally twice daily on days 0, 1, and 2, and on days 7, 8, and 9, before and after the administration of the drug. Treatment was repeated every 21 days unless there was disease progression or unacceptable toxicity.

[0081] Of the 50 patients, 36 received at least one cycle of the study drug without dose changes or dose reductions and were designated as the modified intention to treat (mITT) population. Overall survival in the mITT population in this study was compared to Study ABC-02, an expanded phase 3 randomized controlled trial in patients with unresectable recurrent or metastatic biliary tract cancer (intrahepatic or extrahepatic cholangiocarcinoma, gallbladder cancer, or ampulla cancer). The median overall survival in the mITT population was 20.8 months (95% CI: 13.8-NE), while the median overall survival in the ABC-02 study was 11.7 months (95% CI: 9.5-14.3), indicating an increased survival potential for cholangiocarcinoma patients treated with CX-4945 in combination with cisplatin and gemcitabine. The Kaplan-Meier survival curve for the mITT population is shown in Figure 8.

Claims

1. A pharmaceutical composition for use in a method to treat cancer or refractory cancer in a subject and / or inhibit cancer metastasis, recurrence or progression, or to increase the chances of survival over an appropriate period of time in a subject diagnosed with cancer, comprising CX-4945 or a pharmaceutically acceptable salt or ester thereof, wherein the method comprises the step of administering to a subject a combination of (i) CX-4945 or a pharmaceutically acceptable salt or ester thereof and (ii) an anti-CTLA-4 antibody, wherein CX-4945 has the following structure: 【Chemistry 1】 A pharmaceutical composition having a combination that induces an immune memory against the aforementioned cancer.

2. The pharmaceutical composition according to claim 1, wherein administration increases tumor-specific T cells in the tumor microenvironment of the target.

3. The pharmaceutical composition according to claim 1, wherein the combination of component (i) and component (ii) is administered simultaneously, sequentially, intermittently, or periodically.

4. The pharmaceutical composition according to claim 1, wherein the method comprises at least one administration cycle, the cycle having a duration of 8 weeks or less.

5. The pharmaceutical composition according to claim 4, wherein the cycle has a period of 4 weeks.

6. The pharmaceutical composition according to claim 4, wherein the administration cycle comprises administering component (i) to a subject in an amount and regimen sufficient to achieve a response, and then administering a certain amount of component (ii) to the subject.

7. The pharmaceutical composition according to claim 6, comprising administering component (i) to a subject in an amount and regimen sufficient to achieve a partial or complete response, and then administering a certain amount of component (ii) to the subject.

8. The pharmaceutical composition according to claim 4, wherein component (i) is administered in a daily dose ranging from 25 mg (25 mg / kg) to 2000 mg / kg per kg of body weight of the subject.

9. The pharmaceutical composition according to claim 8, wherein the daily dose is in the range of 50 mg / kg to 200 mg / kg.

10. The pharmaceutical composition according to claim 1, wherein component (ii) is administered in a dose ranging from 1 mg (1 mg / kg) to 20 mg / kg per kg of body weight of the subject.

11. The pharmaceutical composition according to claim 1, wherein the dose of component (ii) is in the range of 8 mg / kg to 15 mg / kg.

12. The pharmaceutical composition according to claim 1, wherein the cancer is glioblastoma, skin cancer, liver cancer, colorectal cancer, glioblastoma, gastric cancer, colorectal cancer, esophageal cancer, lung cancer, pancreatic cancer, renal cell carcinoma, benign prostatic hyperplasia, prostate cancer, ovarian cancer, melanoma, breast cancer, chronic lymphocytic leukemia (CLL), Merkel cell carcinoma, non-Hodgkin lymphoma, acute myeloid leukemia (AML), gallbladder cancer, bile duct cancer, bladder cancer, or uterine cancer.

13. The pharmaceutical composition according to claim 12, wherein the cancer is melanoma, basal cell carcinoma, hepatocellular carcinoma, non-small cell lung cancer (NSCLC), or small cell lung cancer.

14. The pharmaceutical composition according to claim 1, wherein the anti-CTLA-4 antibody is tisilimmab or ipilimumab.

15. The pharmaceutical composition according to claim 1, wherein the cancer is colorectal cancer, colorectal cancer, colon cancer, colon cancer, breast cancer, or breast cancer.

16. The pharmaceutical composition according to claim 1, wherein component (i) is administered twice a day.

17. The pharmaceutical composition according to claim 1, for use in a method for treating refractory cancer in a subject and / or inhibiting metastasis, recurrence or progression of refractory cancer, or for increasing the chances of survival over an appropriate period of time in a subject diagnosed with refractory cancer.

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