Pharmaceutical composition for preventing or treating cancer comprising a 3-ketoacyl COA thiolase inhibitor and a carnitine acylcarnitine carrier inhibitor

Abstract

The present invention relates to a 3-ketoacyl-CoA thiolase inhibitor and a carnitine acylcarnitine carrier. The present invention relates to a pharmaceutical composition and an anti-cancer adjuvant for the prevention or treatment of cancer, which contains an anti-cancer inhibitor. 3-Ketoacyl-CoA thiolase inhibitors and carnitine acylcarnitine carriers of the present invention The composition comprising the inhibitor may be a 3-ketoacyl CoA thiolase inhibitor or a carnitine acyl carnitine thiolase inhibitor. Compared with the use of each nitine carrier inhibitor alone, cancer cell growth, oxygen consumption, and tumor size are significantly reduced, and therefore the combination can be provided as an effective anticancer agent.

Description

Detailed Description of the Invention

[0001] [Technical field] This application claims priority from Korean Patent Application No. 10-2020-0080973, filed on July 1, 2020. and the entire specification is incorporated herein by reference.

[0002] The present invention relates to a pharmaceutical composition and an anticancer adjuvant for preventing or treating cancer, which comprises a 3-ketoacyl CoA thiolase inhibitor and a carnitine acylcarnitine carrier inhibitor.

[0003] [Background technology] While normal cells can grow and suppress regularly and resiliently as needed, cancer cells grow without limit, and are also called tumors, as a mass of undifferentiated cells. These cancer cells infiltrate into surrounding tissues and metastasize to other organs in the body, causing severe pain and eventually death. Despite medical advances, the number of cancer patients in Japan has been steadily increasing, increasing by about 44% in the last 10 years, and the international anti-cancer drug market has also grown, reportedly reaching a scale of about 100 billion dollars per year. It has been reported.

[0004] Anticancer drugs are classified into first-generation chemical anticancer drugs and second-generation targeted anticancer drugs. To overcome the side effects of these drugs, third-generation immune anticancer drugs are being developed. New anticancer drugs have been developed and research is ongoing. However, the biggest problem in cancer treatment today is cancer recurrence. This is not only because cancer mutations are diverse and it is difficult to target a specific cancer, but also because there are many cases where resistance to anticancer drugs used in the treatment of recurrent cancer develops. In the end, even after treatment of the primary cancer, most patients die from metastasis and recurrent cancer. As a result, a strategy of combining anticancer drugs to improve the effectiveness of anticancer drugs has been proposed.

[0005] Trimetazidine, a 3-ketoacyl CoA thiolase (ACAA) inhibitor, has been used as an anti-ischemic (anti-anginal) metabolic agent and as a fatty acid oxidation inhibitor. It is known to have the effect of improving myocardial glucose utilization through inhibition of fatty acid metabolism, and carnitine acylcarnitine carrier (CAC) The inhibitor, omeprazole, is a proton pump inhibitor and is effective in treating gastroesophageal reflux disease, peptic ulcers, erosive esophagitis, and eosinophilic esophagitis. It is known to have results.

[0006] Korean Patent Publication No. 10-2020-0041806 is for a pharmaceutical composition for preventing or treating cancer comprising a malate-aspartate shuttle transport inhibitor and a carnitine acylcarnitine carrier inhibitor, and it is described that the type of carnitine acylcarnitine carrier inhibitor includes trimetazidine or omeprazole.

[0007] However, 3-ketoacyl CoA thiolase (ACAA) inhibitors and carnitine acylcarnitine carrier (CAC) inhibitors No description has been disclosed from studies on whether anticancer effects can be obtained by combining anticancer agents with other anticancer agents.

[0008] [Summary of the Invention] [Problem to be solved by the invention] Therefore, the present inventors have endeavored to provide a combination anticancer drug capable of significantly suppressing cancer cells, and have discovered a combination of a 3-ketoacyl-CoA thiolase inhibitor and a carnitine acylcarnitine carrier. The present invention was completed based on the finding that the cancer cell suppression effect is significantly increased when inhibitors are used in combination, compared with when each is treated alone.

[0009] Therefore, the object of the present invention is to provide a method for the preparation of a 3-ketoacyl CoA thiolase inhibitor and a carnitine acylase inhibitor. The present invention provides a pharmaceutical composition for preventing or treating cancer or an anti-cancer adjuvant, which comprises a monocarnitine carrier inhibitor. [Means for solving the problem]

[0010] In order to achieve the above object, the present invention relates to a 3-ketoacyl CoA thiolase (ACAA) inhibitor and a carnitine acyl ester represented by the following formula (1): Cancer prevention or treatment with carnitine acylcarnitine carrier (CAC) inhibitors or a pharmaceutical composition for treating the same comprising:

[0011] [ka]

[0012] (wherein R1 to R4 are each independently H, C which is optionally substituted or unsubstituted with one or more halogens. 1-6 Optionally substituted or unsubstituted with alkyl, or one or more halogens. I C 1-6 alkoxy, where halogen is selected from the group consisting of F, Cl, Br, and I. It is selected.) The present invention can also provide an anticancer adjuvant comprising a 3-ketoacyl-CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor represented by the above formula (1).

[0013] The ACAA inhibitor can be trimetazidine (KN713), ranolazine (KN715), or a pharma- ceutically acceptable salt thereof. The CAC inhibitor may be omeprazole (KN510), lansoprazole (KN511), pantoprazole (KN512), or a pharma- ceutically acceptable salt thereof.

[0014] The ACAA inhibitor and the CAC inhibitor may be contained in a concentration ratio of 1:100 to 100:1. The ACAA inhibitor and the CAC inhibitor can be administered sequentially or simultaneously.

[0015] The cancer may include one or more selected from the group consisting of colon cancer, lung cancer, gastric cancer, breast cancer, brain cancer, melanoma, glioblastoma, prostate cancer, ovarian cancer, renal cancer, pancreatic cancer, blood cancer and liver cancer. The pharmaceutical composition or the anti-cancer adjuvant may further comprise an additional anti-cancer agent.

[0016] The additional anticancer agent may be irinotecan, paclitaxel, caffecitabine (5-fu), gemcitabine, vemurafenib, or a pharmacologic equivalent thereof. It may be an acceptable salt.

[0017] The present invention can also provide a method for preventing or treating cancer, comprising administering or taking to an individual a composition containing a 3-ketoacyl-CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor represented by the above formula (1) as active ingredients.

[0018] The present invention also provides a cancer prevention or treatment use of a composition containing a 3-ketoacyl-CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor represented by the above formula (1) as active ingredients.

[0019] [Effects of the invention] 3-Ketoacyl-CoA Thiolase Inhibitors and Carnitine Acylcarnitine Carriers of the Present Invention The composition comprising the inhibitor is a 3-ketoacyl CoA thiolase inhibitor or a carnitine acyl carnitine inhibitor. Compared with the case where each nitine carrier inhibitor is used alone, the growth of cancer cells, oxygen consumption, and tumor size are significantly reduced, and therefore the combination can be provided as an effective anticancer drug. [Brief description of the drawings]

[0020] [Figure 1] Figure 1 shows the inhibitory effect of trimetazidine (KN713) and / or omeprazole (KN510) on the proliferation of pancreatic cancer cell lines (MIA PaCa2 and Capan1). It was confirmed that the proliferation of the cell lines was significantly inhibited when trimetazidine or omeprazole was treated in combination compared to when each was treated alone. [Diagram 2] Figure 2 shows the inhibitory effect of treating pancreatic cancer cell lines (Capan2 and BxPC-3) with trimetazidine (KN713) and / or omeprazole (KN510) on their proliferation. It was confirmed that the proliferation of the cell lines was significantly inhibited when trimetazidine or omeprazole was treated in combination compared to when each was treated alone. [Diagram 3] Figure 3 shows the inhibitory effect of trimetazidine (KN713) and / or omeprazole (KN510) on the proliferation of pancreatic cancer cell lines (SNU-213 and SNU-324). It was confirmed that the proliferation of the cell lines was significantly inhibited by the combined treatment with trimetazidine or omeprazole compared to the treatment with either alone. [Figure 4] Figure 4 shows the oxygen consumption of pancreatic cancer cell line MIA PaCa2 treated with trimetazidine (KN713) and / or omeprazole (KN510). It was confirmed that the oxygen consumption was significantly reduced when trimetazidine or omeprazole was treated in combination compared to when each was treated alone. [Diagram 5]Figure 5 shows the change in tumor size when trimetazidine (KN713) was administered to a mouse model implanted with the pancreatic cancer cell line MIA PaCa2. It was confirmed that the tumor size was significantly suppressed as the dose of KN713 increased. [Figure 6] Figure 6 shows the sites of action of trimetazidine and omeprazole. Trimetazidine inhibits 3-ketoacyl CoA thiolase (ACAA1) in peroxisomes, and omeprazole suppresses carnitine acylcarnitine carrier transport in mitochondria. [Figure 7a] FIG. 7a shows the results of analyzing the effect of combined treatment with a CAC inhibitor, KN510 (Omeprazole), and an ACAA inhibitor, KN715 (Ranolazine). [Figure 7b] FIG. 7b shows the results of analyzing the effect of combined treatment with a CAC inhibitor, KN511 (Lansoprazole), and an ACAA inhibitor, KN713 (Trimetazidine). [Figure 7c] FIG. 7c shows the results of analyzing the effect of combined treatment with the CAC inhibitor KN512 (Pantoprazole) and the ACAA inhibitor KN713 (Trimetazidine). [Figure 7d] FIG. 7d shows the results of analyzing the effect of combined treatment with a CAC inhibitor, KN511 (Lansoprazole), and an ACAA inhibitor, KN715 (Ranolazine). [Figure 7e] FIG. 7e shows the results of analyzing the effect of combined treatment with the CAC inhibitor KN512 (Pantoprazole) and the ACAA inhibitor KN715 (Ranolazine). [Figure 8a]FIG. 8 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in colon cancer cell lines. [Figure 8b] FIG. 8 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in colon cancer cell lines. [Figure 9a] FIG. 9 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in renal cancer cell lines. [Figure 9b] FIG. 9 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in renal cancer cell lines. [Figure 10a] FIG. 10 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in liver cancer cell lines. [Figure 10b] FIG. 10 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in liver cancer cell lines. [Figure 11a] FIG. 11 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in breast cancer cell lines. [Figure 11b] FIG. 11 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in breast cancer cell lines. [Figure 12a]FIG. 12 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in ovarian cancer cell lines. [Figure 12b] FIG. 12 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in ovarian cancer cell lines. [Figure 13a] FIG. 13 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in prostate cancer cell lines. [Figure 13b] FIG. 13 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in prostate cancer cell lines. [Figure 14a] FIG. 14 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anti-cancer drug in glioblastoma (GBM) cell lines. [Figure 14b] FIG. 14 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anti-cancer drug in glioblastoma (GBM) cell lines. [Figure 15a] FIG. 15 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in melanoma cell lines. [Figure 15b] FIG. 15 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in melanoma cell lines. [Figure 16a]FIG. 16 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anti-cancer drug in pancreatic cancer (PDAC) cell lines. [Figure 16b] FIG. 16 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anti-cancer drug in pancreatic cancer (PDAC) cell lines. [Figure 17a] FIG. 17 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in gastric cancer cell lines. [Figure 17b] FIG. 17 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in gastric cancer cell lines. [Figure 18a] FIG. 18 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in non-small cell lung cancer (NSCLC) cell lines. [Figure 18b] FIG. 18 shows the results of analyzing triple combination treatment of a CAC inhibitor (KN510: omeprazole), an ACAA inhibitor (KN713: trimetazidine) and an anticancer drug in non-small cell lung cancer (NSCLC) cell lines. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] The present invention will now be described in more detail. As described above, the chemical anticancer drugs conventionally used in cancer treatment have problems such as side effects and a tendency to develop drug resistance, and therefore there is a demand for the development of alternative targeted anticancer therapeutic agents.

[0022] The 3-ketoacyl CoA thiolase (ACAA) inhibitors according to the present invention Carnitine Acylcarnitine Carrier: The composition containing a 3-ketoacyl CoA thiolase (ACAA) inhibitor or a carnitine acylcarnitine carrier (CAC) inhibitor has been shown to inhibit the growth and proliferation of cancer cells, as compared to the use of either a 3-ketoacyl CoA thiolase (ACAA) inhibitor or a carnitine acylcarnitine carrier (CAC) inhibitor alone. Since the oxygen consumption or tumor size is significantly reduced, the composition is effective as a cancer prevention or treatment composition or anti-cancer adjuvant.

[0023] Therefore, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising a 3-ketoacyl-CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor represented by the following formula (1):

[0024] [ka]

[0025] (wherein R1 to R4 are each independently H, C which is optionally substituted or unsubstituted with one or more halogens. 1-6 Optionally substituted or unsubstituted with alkyl, or one or more halogens. I C 1-6 alkoxy, where halogen is selected from the group consisting of F, Cl, Br, and I. It is selected.) The ACAA inhibitor can be trimetazidine (KN713), ranolazine (KN715), or a pharma- ceutically acceptable salt thereof.

[0026] The CAC inhibitor may be omeprazole (KN510), lansoprazole (KN511), pantoprazole (KN512), or a pharma- ceutically acceptable salt thereof. The ACAA inhibitor and the CAC inhibitor may be contained in a concentration ratio of 1:100 to 100:1.

[0027] The ACAA inhibitor and the CAC inhibitor can be administered sequentially or simultaneously. The cancer may be selected from the group consisting of colon cancer, lung cancer, gastric cancer, breast cancer, brain cancer, melanoma, glioblastoma, prostate cancer, ovarian cancer, renal cancer, pancreatic cancer, blood cancer and liver cancer.

[0028] The composition of the present invention can be in various formulations, oral or parenteral.When formulating the composition, one or more buffers (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g., aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrating agents or The formulation may be prepared using a surfactant, diluent or excipient.

[0029] Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. Starch, etc.), calcium carbonate, sucrose The active ingredient is mixed with a solid excipient, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, gelatin, etc. For example, tablets or sugar-coated tablets can be obtained by mixing the active ingredient with a solid excipient, pulverizing it, and adding a suitable adjuvant to process it into a granular mixture.

[0030] In addition to simple excipients, lubricants such as magnesium stearate, talc, etc. are also used. Liquid preparations for oral administration include suspensions, liquid preparations, emulsions, syrups, etc., and in addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, flavorings, preservatives, etc. may be included. In some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, sodium alginate, etc. may be added as a disintegrant, and anti-agglomerating agents, lubricants, wetting agents, flavorings, emulsifiers, preservatives, etc. may further be included.

[0031] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, twin 61, cacaodi, laurindi, glycerol, and gelatin.

[0032] The composition of the present invention can be administered orally or parenterally, and when administered parenterally, it can be administered topically to the skin; They can be formulated according to methods known in the art in the form of injectables to be injected intraperitoneally, rectally, intravenously, intramuscularly, subcutaneously, intrauterine dura or intracerebral blood vessels.

[0033] The above-mentioned injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Examples of suitable carriers for injections include, but are not limited to, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils, including solvents or dispersion media. More preferably, suitable carriers include isotonic solutions such as Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine or sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. To protect the injection from microbial contamination, various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In addition, the injection may further contain an isotonic agent such as sugar or sodium chloride in most cases.

[0034] The compositions of the invention are administered in a pharma- ceutical effective amount, meaning an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to any medical treatment, and including effective dosage levels. The dose can be determined according to factors including the type and severity of the patient's disease, the activity of the drug, its sensitivity to the drug, the administration time, the route of administration and the excretion rate, the duration of treatment, the drugs used simultaneously, and other factors well known in the medical field. The compositions of the present invention can be administered as individual therapeutic agents or in combination with other therapeutic agents, can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered in single or multiple doses. That is, the total effective amount of the compositions of the present invention can be administered to the patient in a single dose, or can be administered by a divided treatment method in which multiple doses are administered over an extended period of time. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect at the minimum amount without side effects, which can be easily determined by those skilled in the art.

[0035] The preferred dose of the composition varies depending on the patient's condition, body weight, degree of disease, drug form, administration route and period, but can be appropriately selected by those skilled in the art. For example, it is 0.0 mg / day. 0.001 to 2,000 mg / kg, more preferably 0.001 to 2,000 mg / kg The dosage may be administered once a day or in several divided doses per day. However, the scope of the present invention is not limited to the above dosage.

[0036] The compositions of the present invention may be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy and methods using biological response modifiers. Therefore, a pharmaceutical composition for preventing or treating cancer comprising a 3-ketoacyl CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor of the present invention is administered to a cancer cell line. When these are used together, a significantly increased cancer cell inhibitory effect can be obtained compared to when each is treated alone. In this case, the control group used for comparing the cancer cell inhibitory effect is a medium treated with a vehicle solvent, and the vehicle can be DW (1%), DMSO (0.1% to 0.2%) or SMSO (0.1%).

[0037] The present invention can also provide an anticancer adjuvant comprising a 3-ketoacyl-CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor represented by the above formula (1).

[0038] The ACAA inhibitor can be trimetazidine (KN713), ranolazine (KN715), or a pharma- ceutically acceptable salt thereof. The CAC inhibitor may be omeprazole (KN510), lansoprazole (KN511), pantoprazole (KN512), or a pharma- ceutically acceptable salt thereof.

[0039] The ACAA inhibitor and the CAC inhibitor may be contained in a concentration ratio of 1:100 to 100:1. The ACAA inhibitor and the CAC inhibitor can be administered sequentially or simultaneously.

[0040] The cancer may be selected from the group consisting of colon cancer, lung cancer, gastric cancer, breast cancer, brain cancer, melanoma, glioblastoma, prostate cancer, ovarian cancer, renal cancer, pancreatic cancer, blood cancer and liver cancer. The anticancer adjuvant of the present invention means all forms for increasing the anticancer effect of anticancer drugs or suppressing or improving the side effects of anticancer drugs.The anticancer adjuvant of the present invention can be administered in combination with various kinds of anticancer drugs or anticancer adjuvant drugs, and even if the anticancer drug is administered at a lower level than the usual dose of anticancer drugs during the combined administration, it can show the same level of anticancer therapeutic effect, so that it can perform safer anticancer treatment.

[0041] The administration route of the anticancer adjuvant can be any common route as long as it can reach the target tissue.The anticancer adjuvant of the present invention can be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, orally, pulmonary, or rectally as desired, but is not limited thereto.In addition, the anticancer adjuvant can be administered by any device that can move the active substance to the target cell.

[0042] The anti-cancer adjuvant of the present invention can be preferably formulated as an anti-cancer adjuvant that contains one or more pharma- ceutically acceptable carriers in addition to the active ingredient for administration.The carrier, excipient or diluent that can be contained in the anti-cancer adjuvant of the present invention includes, but is not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0043] The anticancer adjuvant of the present invention may be a formulation for oral or parenteral administration, and the description of the formulation replaces the description of the formulation of the pharmaceutical composition. [Mode for carrying out the invention] The present invention will be described in more detail below with reference to examples. These examples are merely for the purpose of illustrating the present invention, and it is obvious to those skilled in the art that the scope of the present invention is not limited by these examples.

[0044] [Example 1] SRB analysis 1 Pancreatic cancer (MIA PaCa2, Capan1, Capan2, BxPC-3, SNU-213 or SNU-324) cell lines (100 μl) were seeded into 96-well microtiter plates at densities ranging from 7,500 to 10,000 cells / well depending on the doubling time of each cell line. Meprazole (Omeprazole, KN510) 100 or 200 μM, trimetazidine 1 mM + omeprazole 100 μl of each of 100 μM trimethoprim, 1 mM trimetazidine + 200 μM omeprazole were added to each well, and the plate was incubated in a CO2 incubator, followed by the addition of cold TCA to terminate the assay. Cells were fixed in situ by gently adding 50 μl of cold 50% (w / v) TCA (final concentration: 10% TCA) and incubated at 4 °C for 60 min. The supernatant was discarded, and the plate was washed five times with tap water and air-dried. 100 μl of 0.4% (w / v) sulforhodamine B solution in 1% acetic acid was added to each well, and the plate was left at room temperature for 10 min. After staining, unbound dye was removed by washing five times with 1% acetic acid, and the plate was air-dried. Bound dye was then solubilized with 10 mM trizma base, and absorbance was measured at 515 nm using an automated plate reader. The data were recorded using a

[0045] As a result, as shown in [Figures 1] to [Figures 3], it was confirmed that synergy occurs when trimetazidine or omeprazole is used in combination compared to when each is used alone, and the effect of inhibiting the growth of cancer cell lines is significantly increased. The following [Tables 1] to [Table 3] show the degree of growth inhibition (relative to the control group) for the pancreatic cancer cell lines in [Figures 1] to [Figure 3], respectively.

[0046] [Table 1]

[0047] [Table 2]

[0048] [Table 3]

[0049] [Example 2] Oxygen consumption measurement The oxygen consumption rate (OCR), basal respiration, and ATP production (ATP ... Production) was measured.

[0050] Specifically, cells were plated in MIA PaCa-2 cell culture plates (Seahorse Bioscience, North Billerica). MIA PaCa-2 cells were seeded at 15,000 cells / well (XF96 plates) and incubated in a humidified 37°C incubator with 5% CO2 for 24 hours. Before performing the analysis, the growth medium in the wells was at a minimum concentration of 1:1000, and 170 μl of analysis medium was added to the cells. During calibration of the sensor cartridge, the cell plates were incubated for 60 minutes in a 37°C / non-CO2 incubator before the analysis started. All experiments were performed at 37°C. Each measurement cycle consisted of a 2 minute mixing time and a 4 minute data collection cycle time. Respiratory chain inhibitors (oligomycin, FCCP (carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone), rotenone & antymycin A) (25 μl) were prepared to appropriate concentrations in the cartridge, adjusted to pH 7.4, and then added to each injection port. Three baseline measurements were taken before agent addition and three responses were taken after each addition. OCR data points represent the average absolute rate (pmoles / min) during the measurement cycle. [Table 4 ] indicates the basal respiration rate or ATP production rate in [Figure 4].

[0051] [Table 4]

[0052] As a result, as shown in Figure 4, the combined treatment of trimetazidine and omeprazole significantly reduced oxygen consumption, basal respiration, or ATP, compared to the treatment of either trimetazidine or omeprazole alone. It was confirmed that the production volume was significantly reduced.

[0053] [Example 3] Xenograft tumor models Six to eight week-old Balb / c-nu mice (Orient, Seoul, Korea) were inoculated with MIA PaCa-2 cells (1 × 10 7) The mice were subcutaneously inoculated with 1 ml of the vaccine using a syringe. After one week, the mice were divided into the control group ( Vehicle treatment), KN713 40mg / kg / 100μl treatment group and KN713 80mg / KN713 was administered intraperitoneally once a day (49 days). The primary tumor size was measured weekly using calipers, and the tumor volume was V = (A × B 2 ) / 2 (V = volume (mm 3 ) 、 (A = major axis, B = minor axis). This experiment was conducted in accordance with the Institutional Animal Care and Use Committee (IACUC) of the National Cancer Center Research Institute. The National Cancer Center Research Institute is an AAALAC International approved institution that follows the Laboratory Animal Resources Guide (Protocol: ncc-19-494). [Table 5] shows the tumor size in [Figure 5].

[0054] [Table 5]

[0055] As a result, as shown in FIG. 5, it was confirmed that the tumor size was significantly reduced when treated with trimetazidine or omeprazole in combination compared to when treated with either alone.

[0056] [Example 4] SRB analysis 2 SRB analysis revealed that the pancreatic cancer cell lines SW1990, MIA PaCa2, Panc-1, SU.86.86, and BxPC-3 Carnitine acylcarnitine carrier (CAC) inhibitors and 3-ketoacyl CoA thiolase (3-ketoacyl CoA thiolase) in AsPC-1, SNU-213, and SNU-324 The effect of combined treatment with an ACAA (Actathiolase) inhibitor on cell proliferation was analyzed. The degree of cell proliferation was analyzed relative to the standard value of 100%).

[0057] SRB analysis was performed as follows: 5,000–20,000 cells / well depending on the doubling time of each cell line. Cells (100 μl) were plated in 96 wells at plating densities ranging from 100 μl per well to 100 μl per well. After cell seeding, the microtiter plates were incubated for 24 hours before adding the experimental drugs. Drugs were prepared at the indicated concentrations and added in 100 μl portions to each well. The plates were then incubated in a CO2 incubator. The analysis was then terminated by adding cold TCA. Cells were fixed in situ by gentle addition of 10% TCA (final concentration) and incubated at 4°C for 60 min. The supernatant was discarded and the plate was washed five times with tap water and air-dried. 100 μl of 0.4% (w / v) SRB (Sulforhodamine B) solution in 1% acetic acid was added to each well and the plate was left at room temperature for 10 min. After staining, unbound dye was removed by washing five times with 1% acetic acid and the plate was air-dried. Bound dye was then eluted with 10 mM Tris(OH)2Cl2. The mixture was solubilized with mercury base and the absorbance was recorded at 515 nm using an automated plate reader.

[0058] Figure 7a and Table 6 show the results of the CAC inhibitor KN510 (omeprazole) and the ACAA inhibitor The results are shown in Table 1. The results are shown in Table 1. The effects of combined treatment with KN510 (ranolazine), a drug used to treat glaucoma, were analyzed. The results are shown in Table 1. The control, KN510 100 μM alone, KN510 200 μM alone, KN715 200 μM alone, and KN510 100μM + KN715 200μM (combined treatment), KN510 200μM + KN715 200μM (combined treatment) As shown in Figure 7a and Table 6, the CAC inhibitors inhibited the cytotoxicity of pancreatic cancer cells after 48 hours of treatment. It was confirmed that the combination of KN510 (Omeprazole) and KN715 (Ranolazine), an ACAA inhibitor, significantly inhibited cell proliferation.

[0059] [Table 6]

[0060] Figure 7b and Table 7 show the CAC inhibitor KN511 (Lansoprazole) and the ACAA inhibitor The results of analyzing the effect of combined treatment with KN713 (trimetazidine), a potent anti-cancer drug, are shown below. The results are shown in order: control, KN511 50μM alone, KN511 100μM alone, KN713 2.5mM alone, KN511 50μM + KN713 2.5mM (combined treatment), and KN511 100μM + KN713 2.5mM (combined treatment). As shown in Figure 7b and Table 7, the CAC inhibitor It was confirmed that cell proliferation was significantly suppressed when KN511 (Lansoprazole), an inhibitor of ACAA, was treated in combination with KN713 (Trimetazidine), an inhibitor of ACAA.

[0061] [Table 7]

[0062] Figure 7c and Table 8 show the effect of the CAC inhibitor KN512 (pantoprazole) and ACAA The results of analysis of the effect of combined treatment with the inhibitor KN713 (trimetazidine) are shown below. The results are in order: control, KN512 100μM alone, KN512 200μM alone, and KN713 2.5mM alone. Germany, KN511 50μM + KN713 2.5mM (combined treatment), KN512 100μM + KN713 2.5mM (combined treatment) As shown in Figure 7c and Table 8, the CAC inhibition was observed in pancreatic cancer cell lines treated with 100 mg / kg of 10 ... It was confirmed that cell proliferation was significantly suppressed when the cytotoxic agent KN512 (pantoprazole) and the ACAA inhibitor KN713 (trimetazidine) were treated in combination.

[0063] [Table 8]

[0064] Figure 7d and Table 9 show the CAC inhibitor KN511 (Lansoprazole) and the ACAA inhibitor The results are shown below. The results are based on the analysis of the effect of combined treatment with the anti-inflammatory drug KN715 (ranolazine). The results are listed in order: control, KN511 50μM alone, KN511 100μM alone, KN715 200μM alone, KN511 50μM + KN715 200μM (combined treatment), and KN511 100μM + KN715 200μM (combined treatment). ) was treated with pancreatic cancer cell lines for 48 hours. As shown in Figure 7d and Table 9, CAC inhibition It was confirmed that cell proliferation was significantly suppressed when the cytotoxic agent KN511 (Lansoprazole) and the ACAA inhibitor KN715 (Ranolazine) were treated in combination.

[0065] [Table 9]

[0066] Figure 7e and Table 10 show the effect of the CAC inhibitor KN512 (pantoprazole) and ACAA The results of analysis of the effect of combined treatment with the inhibitor KN715 (ranolazine) are shown. The results are as follows: control, KN512 100μM alone, KN512 200μM alone, KN715 200μM alone, KN512 100μM + KN715 200μM (combined treatment), and KN512 200μM + KN715 200μM (combined treatment) on pancreatic cancer cell lines for 48 hours. As shown in Figure 7e and Table 10, the CAC inhibitor It was confirmed that cell proliferation was significantly suppressed when the agent KN512 (pantoprazole) and the ACAA inhibitor KN715 (ranolazine) were treated in combination.

[0067] [Table 10]

[0068] [Example 5] SRB analysis 3 SRB analysis (n=3) demonstrated that the carnitine acylcarnitine carrier (CAC) inhibitor (KN510: omeprazole) and the 3-ketoacyl CoA thiolase (ACAA) inhibitor (KN713: trimetazidine) were effective in preventing and treating the inflammatory bowel disease in cancer cell lines. ) and triple combination treatments of anticancer drugs were analyzed (see Table 11).

[0069] [Table 11] TIFF0007669053000014.tif218170

[0070] SRB assays were performed as follows: Cells (100 μl) containing 5,000–40,000 cells / well (cells / well) depending on the doubling time of each cell line were incubated in 96-well microtiter plates. After 24 h, drugs (100 μl per well) were added to each well and the cultures were incubated at 37°C for 48 h. The cells were then soaked in 50% TCA ( Ink the plates at 4°C for at least 1 h or up to 3 h. After incubation, remove the liquid from the plate, rinse five times with water, and incubate at room temperature (RT) for approximately 12–24 hours. The fixed cells were stained with 0.4% SRB (100 μl per well) for 10 min at room temperature. Afterwards, the plates were washed three times with 1% glacial acetic acid and dried at room temperature for about 12-24 hours. SRB stain was dissolved in 10 mM trizma base and the absorbance was read at 515 nm. The effect of the drugs was evaluated by GI 50 (50% growth inhibition), TGI (total growth inhibition) or LC 50 (lethal concentration). GI 50 Cell proliferation is the maximum concentration at which 50% inhibition occurs, and IC 50 is the concentration of drug that inhibited in vitro enzyme activity by 50% in this study.

[0071] As a result of the analysis, as shown in Figures 8 to 18 and Tables 12 to 67, it was confirmed that the cancer cell lines were inhibited by triple combined treatment with a carnitine acylcarnitine carrier (CAC) inhibitor (KN510: omeprazole), a 3-ketoacyl CoA thiolase (ACAA) inhibitor (KN713: trimetazidine) and an anticancer drug. It was.

[0072] [Table 12]

[0073] [Table 13]

[0074]

Table 14

[0075]

Table 15

[0076]

Table 16

[0077]

Table 17

[0078]

Table 18

[0079]

Table 19

[0080]

Table 20

[0081]

Table 21

[0082]

Table 22

[0083]

Table 23

[0084]

Table 24

[0085]

Table 25

[0086]

Table 26

[0087]

Table 27

[0088]

Table 28

[0089]

Table 29

[0090]

Table 30

[0091]

Table 31

[0092]

Table 32

[0093]

Table 33

[0094]

Table 34

[0095]

Table 35

[0096]

Table 36

[0097]

Table 37

[0098]

Table 38

[0099]

Table 39

[0100]

Table 40

[0101]

Table 41

[0102]

Table 42

[0103]

Table 43

[0104]

Table 44

[0105]

Table 45

[0106]

Table 46

[0107]

Table 47

[0108]

Table 48

[0109]

Table 49

[0110]

Table 50

[0111]

Table 51

[0112]

Table 52

[0113]

Table 53

[0114]

Table 54

[0115]

Table 55

[0116]

Table 56

[0117]

Table 57

[0118]

Table 58

[0119]

Table 59

[0120]

Table 60

[0121]

Table 61

[0122]

Table 62

[0123]

Table 63

[0124]

Table 64

[0125]

Table 65

[0126]

Table 66

[0127]

Table 67

Claims

1. A pharmaceutical composition for preventing or treating cancer, comprising a 3-ketoacyl CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor represented by the following formula 1: A pharmaceutical composition for preventing or treating cancer, wherein the ACAA inhibitor is trimetazidine, ranolazine, or a pharma- ceutical acceptable salt thereof: 【Chemistry 1】 Here, R 1 ~R 4 each independently represents H, C optionally substituted or unsubstituted with one or more halogens １－６ C optionally substituted or unsubstituted with alkyl, or one or more halogens １－６ alkoxy, where the halogen is selected from the group consisting of F, Cl, Br, and I.

2. The pharmaceutical composition for preventing or treating cancer according to claim 1 , wherein the CAC inhibitor is omeprazole, lansoprazole, pantoprazole, or a pharma- ceutically acceptable salt thereof.

3. 3. The pharmaceutical composition of claim 1 or 2, wherein the ACAA inhibitor and the CAC inhibitor are contained in a concentration ratio of 1:100 to 100:

1.

4. The pharmaceutical composition of claim 1 or 2, wherein the ACAA inhibitor and the CAC inhibitor are administered sequentially or simultaneously.

5. The pharmaceutical composition according to claim 1 or 2, characterized in that the cancer is one or more cancers selected from the group consisting of colorectal cancer, lung cancer, gastric cancer, breast cancer, brain cancer, melanoma, glioblastoma, prostate cancer, ovarian cancer, renal cancer, pancreatic cancer, blood cancer and liver cancer.

6. 3. The pharmaceutical composition of claim 1 or 2, further comprising an additional anti-cancer agent.

7. 7. The pharmaceutical composition of claim 6, wherein the additional anticancer agent is irinotecan, paclitaxel, caffecitabine (5-fu), gemcitabine, vemurafenib, or a pharmaceutically acceptable salt thereof.

8. An anti-cancer adjuvant comprising a 3-ketoacyl CoA thiolase (ACAA) inhibitor and a carnitine acylcarnitine carrier (CAC) inhibitor represented by the following formula 1:

1. An anti-cancer adjuvant, wherein the ACAA inhibitor is trimetazidine, ranolazine, or a pharma- ceutically acceptable salt thereof: 【Chemistry 2】 Here, R 1 ~R 4 each independently represents H, C optionally substituted or unsubstituted with one or more halogens １－６ C optionally substituted or unsubstituted with alkyl, or one or more halogens １－６ alkoxy, where the halogen is selected from the group consisting of F, Cl, Br, and I.

9. 9. The anticancer adjuvant of claim 8, wherein the CAC inhibitor is omeprazole, lansoprazole, pantoprazole, or a pharma- ceutically acceptable salt thereof.

10. The anticancer adjuvant according to claim 8 or 9, wherein the ACAA inhibitor and the CAC inhibitor are contained in a concentration ratio of 1:100 to 100:

1.

11. The anticancer adjuvant of claim 8 or 9, wherein the ACAA inhibitor and the CAC inhibitor are administered sequentially or simultaneously.

12. The anticancer adjuvant according to claim 8 or 9, characterized in that the cancer is at least one cancer selected from the group consisting of brain cancer, melanoma, glioblastoma, prostate cancer, ovarian cancer, renal cancer, pancreatic cancer, blood cancer and liver cancer.

13. The anti-cancer adjuvant according to claim 8 or 9, further comprising an additional anti-cancer drug.

14. The anticancer adjuvant according to claim 13, wherein the additional anticancer drug is irinotecan, paclitaxel, caffecitabine (5-fu), gemcitabine, vemurafenib, or a pharmaceutically acceptable salt thereof.

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