Compositions for combined administration containing 2,3,5-substituted thiophene compounds

Abstract

The present invention relates to a composition for combined administration containing a 2,3,5-substituted thiophene compound, which has superior inhibitory activity against cancer cells associated with leukemia compared to treatment with a 2,3,5-substituted thiophene compound or an anticancer agent alone, and can be usefully used for the prevention, amelioration, or treatment of leukemia.

Description

[Technical Field]

[0001] The present invention relates to co-administration compositions comprising 2,3,5-substituted thiophene compounds. [Background technology]

[0002] Although the development of modern medicine has enabled the treatment and prevention of many diseases, cancer remains one of the most difficult diseases to treat. Cancer is currently the leading cause of death and is showing a continuous increase in incidence.

[0003] Cancer treatment methods include chemotherapy, surgery, and / or radiation therapy. Among these, chemotherapy, which uses anticancer drugs, is the most widely used method for treating cancer. Currently, approximately 60 different anticancer drugs are in clinical use, and new anticancer drugs are continuously being developed as knowledge about cancer development and the characteristics of cancer cells improves. However, most anticancer drugs currently in clinical use often have side effects such as nausea, vomiting, oral and small intestinal ulcers, diarrhea, hair loss, and / or bone marrow suppression, which causes reduced production of active ingredients in the blood. For example, mitomycin C is known to cause renal failure, and adriamycin is known to cause bone marrow suppression. In particular, cisplatin, the most effective anticancer drug developed to date, is widely used to treat testicular cancer, ovarian cancer, lung cancer, head and neck cancer, bladder cancer, stomach cancer, and cervical cancer. However, it has serious side effects, such as hematopoietic toxicity (e.g., anemia), gastrointestinal toxicity (e.g., vomiting and nausea), nephrotoxicity (e.g., renal tubule damage), hearing loss, electrolyte imbalance, shock, and peripheral nerve abnormalities.

[0004] Recently developed new, expensive targeted anticancer drugs with excellent safety have shown efficacy in treating many cancer patients, but they have only shown efficacy in small patient groups identified through companion diagnostics. There is an urgent need for precision medicine, which enables individualized treatment for various cancers.

[0005] Leukemia is a general term for diseases in which white blood cells grow like tumors. Leukemia can be divided into myeloid leukemia and lymphocytic leukemia depending on the white blood cell of origin, and into acute leukemia and chronic leukemia depending on the rate of progression. The clinical manifestations of leukemia vary depending on the type of disease and the nature of the affected cells. Lymphocytic leukemia occurs in lymphoid blood cells, myeloid leukemia occurs in myeloid blood cells, and chronic myeloid leukemia occurs when cells in the maturation stage mutate. Acute myeloid leukemia is caused by damage to myeloid parent cells, which begin to differentiate at a relatively early stage in the hematopoietic process.

[0006] Therefore, the present inventors conducted research to find a dosage form that is effective in treating leukemia, and completed the present invention. Summary of the Invention [Problem to be solved by the invention]

[0007] One object of the present invention is to provide a pharmaceutical composition for preventing or treating leukemia, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof and an anticancer agent: [ka] .

[0008] Another object of the present invention is to provide a method for treating leukemia, which comprises administering the pharmaceutical composition to a patient suffering from leukemia.

[0009] Another object of the present invention is to provide a use of a compound represented by formula 1 for the prevention or treatment of leukemia. [ka] .

[0010] Another object of the present invention is to provide a use of a compound represented by Chemical Formula 1 for the manufacture of a pharmaceutical composition for the prevention or treatment of leukemia. [ka] . [Means for solving the problem]

[0011] In order to achieve the above object, one aspect of the present invention provides a pharmaceutical composition for preventing or treating leukemia, comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof and an anticancer agent: [ka] .

[0012] In one embodiment of the present invention, the anti-cancer agent may be a bcl2 inhibitor.

[0013] In one embodiment of the present invention, the anticancer drug may be one or more of cytarabine, venetoclax, and azacitidine. One aspect of the present invention provides a method for treating leukemia, comprising administering the pharmaceutical composition to a patient with leukemia.

[0014] Another aspect of the present invention provides the use of a compound of formula 1 for the prevention or treatment of leukemia. [ka] .

[0015] Another aspect of the present invention provides the use of a compound of formula 1 for the manufacture of a pharmaceutical composition for the prevention or treatment of leukemia. [ka] . [Effects of the Invention]

[0016] A composition for preventing, ameliorating, or treating leukemia, comprising a 2,3,5-substituted thiophene compound and an anticancer agent according to an embodiment of the present invention, has superior inhibitory activity against cancer cells associated with leukemia compared to treatment with the 2,3,5-substituted thiophene compound or the anticancer agent alone, and can be usefully used for preventing, ameliorating, or treating leukemia. [Brief explanation of the drawings]

[0017] [Figure 1] These results confirm the optimal dosage when venetoclax is administered alone. [Figure 2] These are the results of confirming the optimal dosage when azacitidine is administered alone. [Figure 3] These results confirm the optimal dosage when cytarabine is administered alone. [Figure 4] The results confirm the effect of combined administration of a compound according to one embodiment of the present invention and venetoclax. [Figure 5] The results confirm the effect of combined administration of a compound according to one embodiment of the present invention and venetoclax. [Figure 6] 1 shows the results of confirming the effect of combined administration of a compound according to one embodiment of the present invention and azacitidine. [Figure 7] 1 shows the results of confirming the effect of combined administration of a compound according to one embodiment of the present invention and cytarabine. DETAILED DESCRIPTION OF THE INVENTION

[0018] One aspect of the present invention provides a pharmaceutical composition for preventing or treating leukemia, comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof and an anticancer agent: [ka] .

[0019] The compound represented by Chemical Formula 1, which is contained as an active ingredient in the pharmaceutical composition of the present invention, is (S)-5-((3-fluorophenyl)ethynyl)-N-(piperidin-3-yl)-3-ureidothiophene-2-carboxamide.

[0020] In one embodiment of the present invention, the anticancer drug may be a bcl2 inhibitor. The bcl2 inhibitor refers to a substance that suppresses the inactivation of Bcl2 (B-cell lymphoma 2), a protein that regulates cell death by inducing pro-apoptosis and suppressing anti-apoptosis, by inhibiting its expression. The bcl2 inhibitor may be one or more of cytarabine, venetoclax, and azacitidine.

[0021] Cytarabine (also known as cytarabine) is a cytotoxic anticancer drug that inhibits the synthesis of DNA, which is necessary for cell survival, and is known to be used alone or in combination to treat leukemia, including acute myeloid leukemia, and various other cancers.

[0022] Venetoclax (Venclexta® or Venclyxto®) is a substance used to treat acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), and other diseases.

[0023] Azacitidine (also known as 5-azacytidine, azacitidine, or 5-azcitidine) is known to inhibit protein synthesis by penetrating into RNA and DNA via the nucleic acid biosynthesis pathway in actively dividing cells, thereby exhibiting cytocidal effects.

[0024] In one embodiment of the present invention, the cytarabine is a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof in a treatment ratio of 1:1 to 1:6000; The venetoclax is a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof, and the ratio of the amount of the compound represented by Chemical Formula 1 to the amount of the venetoclax is 1:0.0001 to 1:100. The azacitidine may be contained in a ratio of the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof in a treatment amount of 1:0.02 to 1:14.

[0025] The pharmaceutical composition for preventing or treating leukemia of the present invention has been confirmed to have an excellent synergistic effect, specifically, an excellent effect in the treatment of leukemia, by combining the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof with an anticancer agent.

[0026] The pharmaceutical compositions of the present invention may be prepared in unit dose form or in multi-dose containers by formulating them with pharmaceutically acceptable carriers in a manner that can be easily carried out by a person of ordinary skill in the art to which the invention pertains.

[0027] The pharmaceutically acceptable carriers are those commonly used in formulations, including, but not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above ingredients, the pharmaceutical composition of the present invention may additionally contain lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (22nd ed., 2013).

[0028] In the present invention, the content of the additives contained in the pharmaceutical composition is not particularly limited, and can be appropriately adjusted within the content range used in ordinary formulations.

[0029] The compositions of the present invention can be formulated into injectable dosage forms such as aqueous solutions, suspensions, emulsions and the like, pills, capsules, granules or tablets.

[0030] The pharmaceutical compositions of the present invention may be for oral administration, and non-limiting examples of formulations for oral administration include tablets, troches, lozenges, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs.

[0031] The pharmaceutical composition of the present invention may also be for parenteral administration, and non-limiting examples of parenteral formulations include injection solutions, suppositories, powders for respiratory inhalation, aerosols for spraying, ointments, powders for application, oils, creams, etc.

[0032] The preferred dosage of the pharmaceutical composition of the present invention varies depending on the patient's condition and weight, age, sex, health condition, dietary habits, properties of the formulation, severity of the disease, administration time, administration method, administration period or interval, excretion rate, and drug form, and can be appropriately selected by those skilled in the art. For example, it may be in the range of about 0.1 to 10,000 mg / kg, but is not limited thereto, and can be administered once or several times a day.

[0033] In the present invention, the pharmaceutical composition may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the intended method.

[0034] The combination of the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof with an anticancer drug may be prepared not only in a single dosage form, but also in separate dosage forms and administered together or at different times.

[0035] When prepared into a single dosage form, the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof and the anticancer agent may be mixed together, or the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof and the anticancer agent may be compartmentalized in a single dosage form.

[0036] When prepared as separate dosage forms, the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof and the anticancer agent may be formulated into different forms in consideration of their individual dosage amounts, administration routes, administration times, etc., depending on the properties of the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof and the anticancer agent.

[0037] In one embodiment of the present invention, the compound represented by Chemical Formula 1, its stereoisomer, its pharmaceutically acceptable salt, its hydrate or its solvate may be administered parenterally or orally, preferably orally.

[0038] The pharmaceutically effective amount and effective dosage of the pharmaceutical composition of the present invention vary depending on the formulation method, administration mode, administration time and / or administration route of the pharmaceutical composition, etc., and a person skilled in the art can easily determine and prescribe an effective dosage for the intended treatment.

[0039] The pharmaceutical composition of the present invention may be administered once a day or in divided doses several times a day.

[0040] Another aspect of the present invention provides a method for treating leukemia, comprising administering the pharmaceutical composition to a patient suffering from leukemia.

[0041] A pharmaceutical composition according to one embodiment of the present invention comprises a compound represented by Chemical Formula 1 and an anticancer drug as active ingredients. Therefore, before administering the pharmaceutical composition of the present invention, a companion diagnosis step of selecting a patient group that will respond to the compound represented by Chemical Formula 1 and the anticancer drug may be further included, which is carried out using a leukemia diagnostic method known in the art. Meanwhile, the term "companion diagnosis" as used in the present invention refers to a diagnosis for predicting a patient's responsiveness to a specific drug treatment.

[0042] Another aspect of the present invention provides use of a compound represented by Chemical Formula 1 for the prevention or treatment of leukemia, or use of a compound represented by Chemical Formula 1 for the manufacture of a pharmaceutical composition for the prevention or treatment of leukemia. [Example]

[0043] Hereinafter, one or more specific examples will be described in more detail through examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

[0044] Experimental Example 1: Materials and Methods The experimental materials and equipment used are as follows: [Table 1]

[0045] Experimental Example 2: Confirmation of optimal dosage of individual anticancer drugs Experimental Example 2-1. Confirmation of Venetoclax Dosage The leukemia cell lines prepared were BDCM, HL-60, MolM13, MolM14, MV4-11, THP-1, and U937, and the administered drug was venetoclax.

[0046] In the first and second experiments, the cell lines were tested three times with venetoclax doses reduced by half from 100uM, in the third experiment, the BDCM cell line was tested three times with venetoclax doses reduced by half from 100uM, and in the HL-60, MolM13, MolM14, and MV4-11 cell lines, the HL-60, MolM13, MolM14, and MV4-11 cell lines were tested three times with venetoclax doses reduced by half from 2.5uM, and in the fourth experiment, the HL-60, MolM13, MolM14, and MV4-11 cell lines were tested three times with venetoclax doses reduced by half from 2.5uM.

[0047] Specifically, in the first and second experiments, 50 μl of 10 mM venetoclax was dissolved in 450 μl of solvent to prepare a 10x solution. 250 μl of this solution was serially diluted in half, and 250 μl was filled into an 8E-tube. 80 μl of cell lines were then dispensed into 96-well plates. 10 μl of cell lines were then dispensed using JANUS and cultured under CO2 conditions for 72 hours. After the cell titer glo assay, IC 50 was calculated.

[0048] In the third experiment, 15 μl of 10 mM venetoclax was dissolved in 135 μl of solvent to prepare a 10x solution for the BDCM cell line. 100 μl of this solution was serially diluted in half, and 100 μl was filled into an 8E-tube, after which the cell line was dispensed into a 96-well plate. 25 μl of the cell line was then dispensed using JANUS and cultured under CO2 conditions for 72 hours. After performing the cell titer glo assay, IC 50The IC was calculated. Then, 1 μl of 10 mM venetoclax was dissolved in 399 μl of solvent to prepare a 10x solution for the HL-60, MolM13, MolM14, and MV4-11 cell lines. 200 μl of this solution was serially diluted in half, and 200 μl was filled into an 8E-tube. The cell lines were then dispensed into a 96-well plate. 60 μl of the cell lines were then dispensed using JANUS and cultured under CO2 conditions for 72 hours. After the Celltiter glo assay, the IC was calculated. 50 was calculated.

[0049] In the fourth experiment, the same procedure as in the third experiment was carried out on the HL-60, MolM13, MolM14, and MV4-11 cell lines.

[0050] As a result of the above experiment, as can be seen from Figure 1 and Table 2, the IC 50 Venetoclax demonstrated an IC50 of less than 1uM in HL-60, MolM13, MolM14, and MV4-11 leukemia cancer cell lines. 50 confirmed. [Table 2]

[0051] Experimental Example 2 - Confirmation of the dosage of 2.5-azacytidine The leukemia cell lines prepared were BDCM, HL-60, MolM13, MolM14, MV4-11, THP-1, and U937, and the drug administered was 5-azacytidine.

[0052] In the first and second experiments, the cell line was treated with 5-azacytidine starting from 100 μM, and the concentration was reduced by half three times. Specifically, in the first and second experiments, 15 μl of 20 mM 5-azacytidine was dissolved in 285 μl of solvent to prepare a 10x solution. 250 μl of this solution was serially diluted in half, and 250 μl was filled into an 8E-tube. 80 μl of the cell line (3,000 cells) was then dispensed into a 96-well plate. 10 μl of the cell line was then dispensed using JANUS and cultured for 72 hours under CO2 conditions. After the cell titer glo assay was performed, IC 50 was calculated.

[0053] As a result, as can be seen from Figure 2 and Table 3, 50 got the value. [Table 3]

[0054] Experimental Example 2-3. Confirmation of Cytarabine Dosage The leukemia cell lines prepared were BDCM, HL-60, MolM13, MolM14, MV4-11, THP-1, and U937, and the drug administered was cytarabine. For the first and second experiments, the cell line was treated with 50,000 μM cytarabine, which was then reduced in half three times. Specifically, for the first and second experiments, 10 μL of 50 mM cytarabine was dissolved in 490 μL of solvent to prepare a 10x solution. 250 μL of this solution was serially diluted in half, and 250 μL was filled into an 8E-tube. 80 μL of the cell line (3,000 cells) was then dispensed into a 96-well plate. 10 μL of the cell line was then dispensed using JANUS and cultured for 72 hours under CO2 conditions. After the cell titer glo assay, IC 50 was calculated.

[0055] As a result, as can be seen from Figure 3 and Table 4, 50 got the value. [Table 4]

[0056] Experimental Example 3: Confirmation of combined administration ability Experimental Example 3-1. Confirmation of the efficacy of combined administration of the compound of formula I and venetoclax The leukemia cell lines prepared were BDCM, HL-60, MolM13, MolM14, MV4-11, THP-1, and U937, and the drugs administered were PHI-101 and venetoclax.

[0057] Specifically, in the first and second experiments, 1 μl of 1 mM venetoclax was dissolved in 999 μl of solvent to prepare a 10x solution. 500 μl of this solution was serially diluted in half, and 500 μl was filled into an 8E-tube. 90 μl of cell lines were then dispensed into 96-well plates. 10 μl of cell lines were then dispensed using JANUS and pre-treated for 30 minutes under CO2 conditions. PHI-101 solution was prepared as shown in Table 5 below, and 10 μl of this solution was added to the plate. After 72 hours of incubation under CO2 conditions, the Celltiter glo assay was performed, and IC 50 was calculated. [Table 5]

[0058] In the third and fourth experiments, 5 μl of 50 mM PHI-101 was dissolved in 495 μl of solvent to prepare a 10x solution. 250 μl of this solution was serially diluted in half, and 250 μl was filled into an 8E-tube. 60 μl of cell lines were then dispensed into 96-well plates. 10 μl of cell lines were then dispensed using JANUS and pre-treated for 30 minutes under CO2 conditions. Venetoclax solutions were prepared as shown in Table 6 below, and 10 μl of each solution was added to the plates. After 72 hours of incubation under CO2 conditions, a Celltiter glo assay was performed, and IC 50 was calculated. [Table 6]

[0059] The results obtained were then summarized and compared with those obtained when venetoclax or PHI-101 was administered alone.

[0060] As a result, as can be seen from Table 7 and Figures 4 and 5, the combined administration of venetoclax and PHI-101 in each leukemia cancer cell line was found to be 1.63 to 282,8000 times more effective than the administration of venetoclax or PHI-101 alone. [Table 7]

[0061] Experimental Example 3-2. Confirmation of the efficacy of combined administration of the compound of formula I and azacitidine The leukemia cell lines used were BDCM, HL-60, MolM13, MolM14, MV4-11, THP-1, and U937, and the drugs used were 5-azacytidine and PHI-101. Specifically, in the first and second experiments, 5 μl of 50 mM PHI-101 was dissolved in 495 μl of solvent to prepare a 10x solution for BDCM, HL-60, THP-1, and U937 cell lines. 250 μl of this solution was serially diluted in half, and 250 μl was filled into an 8E-tube. 60 μl of the cell line (3,000 cells) was dispensed into a 96-well plate. 10 μl of the cell line was then dispensed using JANUS and pretreated for 30 minutes under CO2 conditions. 5-azacytidine solution was prepared as shown in Table 8 below, and 10 μl of this was added to the plate. After 72 hours of incubation under CO2 conditions, the Celltiter glo assay was performed, and the IC 50 was calculated. [Table 8]

[0062] For MolM13, MolM14, and MV4-11 cell lines, 1 μl of 1 mM PHI-101 was dissolved in 999 μl of solvent to prepare a 10x solution. 250 μl of this solution was serially diluted in half, and 250 μl was filled into an 8E-tube. 50 μl of the cell line (3,000 cells) was dispensed into a 96-well plate. 10 μl of the cell line was then dispensed using JANUS and pretreated for 30 minutes under CO2 conditions. 5-azacytidine solution was prepared as shown in Table 9 below, and 10 μl of this was added to the plate. After 72 hours of culture under CO2 conditions, the Celltiter glo assay was performed, and IC 50 was calculated. [Table 9]

[0063] The results obtained were then summarized and compared with those obtained when 5-azacytidine or PHI-101 was administered alone.

[0064] As a result, as can be seen from Table 10 and Figure 6, the combined administration of 5-azacytidine and PHI-101 was 2 to 3,180 times more effective than the administration of 5-azacytidine or PHI-101 alone. In HL-60 and U937 leukemia cell lines, PHI-101 treatment showed less than 50% inhibitory activity at the lowest concentration. [Table 10]

[0065] Experimental Example 3-3. Confirmation of the efficacy of combined administration of the compound of formula I and cytarabine The leukemia cell lines used were BDCM, HL-60, MolM13, MolM14, MV4-11, THP-1, and U937, and the drugs used were cytarabine and PHI-101. Specifically, in the first and second experiments, 5 μl of 50 mM PHI-101 was dissolved in 495 μl of solvent to prepare a 10x solution for BDCM, HL-60, THP-1, and U937 cell lines. 250 μl of this solution was serially diluted in half, and 250 μl was filled into an 8E-tube. 60 μl of the cell line (3,000 cells) was dispensed into a 96-well plate. Then, 10 μl of the cell line was dispensed using JANUS and pretreated for 30 minutes under CO2 conditions. Next, cytarabine solution was prepared as shown in Table 11 below, and 10 μl of this was added to the plate. After 72 hours of incubation under CO2 conditions and the Celltiter glo assay, IC 50 was calculated. [Table 11]

[0066] For MolM13, MolM14, and MV4-11 cell lines, 1 μl of 1 mM CoA PHI-101 was dissolved in 999 μl of solvent to prepare a 10x solution. 250 μl of this solution was serially diluted in half, and 250 μl was filled into an 8E-tube. 50 μl of the cell line (3,000 cells) was dispensed into a 96-well plate. 10 μl of the cell line was then dispensed using JANUS and pretreated for 30 minutes under CO2 conditions. Cytarabine solution was prepared as shown in Table 12 below, and 10 μl of this was added to the plate. After 72 hours of incubation under CO2 conditions and the Celltiter glo assay, IC 50 was calculated. [Table 12]

[0067] The results obtained were then summarized and compared with those obtained when cytarabine or PHI-101 was administered alone. As a result, as can be seen from Table 13 and Figure 7, the combined administration of cytarabine and PHI-101 was found to be 2 to 19.2 times more effective than the single administration of cytarabine or PHI-101. [Table 13]

[0068] The present invention has been described above with a focus on preferred embodiments. Those skilled in the art will understand that the present invention can be realized in modified forms without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a restrictive perspective. The scope of the present invention is defined by the claims, not the above description, and all variations within the scope of the claims should be construed as being within the scope of the present invention.

Claims

1. A pharmaceutical composition for preventing or treating leukemia, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof and an anticancer agent, wherein the anticancer agent is one or more of venetoclax and azacitidine: 【Chemical 1】 。

2. Use of a compound represented by Chemical Formula 1 and an anticancer agent for the manufacture of a pharmaceutical composition for the prevention or treatment of leukemia, wherein the anticancer agent is one or more of venetoclax and azacitidine. 【Chemistry 2】 。

Images