KR20150117504A - Pharmaceutical compositions for the treatment or prevention of cancers comprising idebenone - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating or preventing cancers by inhibiting a calcium-activated chloride channel (CaCC) and/or anoctamin 1 (ANO1) and, particularly, to a pharmaceutical composition comprising an idebenone compound. The pharmaceutical composition provides effects in treating or preventing prostate cancer, pancreatic cancer, colorectal cancer, lung cancer, head and neck cancer, esophageal cancer, digestive organ cancer, breast cancer, and renal cancer.

Description

[0001] PHARMACEUTICAL COMPOSITIONS FOR THE TREATMENT OR PREVENTION OF CANCERS COMPRISING IDEBENONE [0002]

The present invention relates to a pharmaceutical composition for the treatment or prevention of cancer comprising an idebone compound as an active ingredient.

Idebenone is a synthetic derivative of coenzyme Q10, which is already being used / developed for the treatment of Friedreich ataxia or mitochondrial and neuromuscular diseases. It is used for improving hearing loss due to external damage, improving optic neuritis, The effect of suppressing aging, moisturizing skin, and improving wrinkles due to an increase in collagen synthesis.

The calcium-activated chloride channel (CaCC) is widely expressed in various cells and tissues and regulates physiological functions such as epithelial body fluid secretion, smooth muscle contraction, sensory neuron signaling, and cell growth. do. Until recently, it was found that the CaCC gene was not known, but in 2008, ANO1 / TMEM16A was found to be CaCC, and various studies have revealed that ANO1 has been shown to be involved in bronchial, intestinal, luteal, smooth muscle, Cancer, kidney cancer, and many other types of tumor cells have been shown to have a variety of physiological functions.

Anoctamin 1 (ANO1) is located in chromosome band 11q13 and overexpressed in various cancer cells such as head and neck cancer, breast cancer and prostate cancer. Recent studies have shown that ANO1 is involved in cell growth, migration, cancer cell growth, and cancer development. For example, inhibition of ANO1 expression in prostate cancer cells (PC3) inhibited cell growth, metastasis, infiltration, and tumor development in the mouse model. T16A _inh -A01, an ANO1 inhibitor, (ICC) and pancreatic cancer cells (CFPAC1). In addition, inhibition of ANO1 expression in breast cancer cells has been reported to inhibit the growth of breast cancer cells, induce the death of cancer cells, and inhibit tumor growth in the xenograft model. In addition, inhibition of ANO1 has been shown to inhibit HNSCC, ESCC, and EGFR and calmodulin-dependent protein kinase 2 (CAMK2) signaling in breast cancer cells It turned out.

Recent studies suggest that the inhibition of the pharmacological activity of the ANO1 ion channel is likely to be a therapeutic agent for head and neck cancer, esophageal cancer, gastrointestinal cancer, breast cancer and prostate cancer, which overexpress ANO1. Since the ANO1 channel has recently been identified, CaCC _inh -A01, Tannic acid, T16A _inh -A01, digallic acid, dichlorophen, benzbromarone, and N - (4-methoxy) -2-naphthyl-5-ntroanthranilic acid (MONNA) Although some ANO1 inhibitors have been discovered, the pharmacological mechanisms and properties of these inhibitors have not yet been determined.

The present inventors have made efforts to develop a therapeutic agent for effectively inhibiting various tumors, particularly cancer cells overexpressing CaCC / ANO1. In particular, we sought to develop anticancer drugs by identifying CaCC / ANO1 inhibitors that can be repositioned through screening using existing drugs, drugs under development, and natural products.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer.

In order to solve the above problems, the present inventors used a cell-based high-speed mass screening method to search about 2,500 compounds (including licensed drugs and drugs under development and natural products) and found that idebenone effectively inhibited CaCC / ANO1 .

According to one aspect of the present invention, there is provided a pharmaceutical composition for treating or preventing cancer comprising an effective amount of an ibebenone compound represented by the following formula (1).

[Chemical Formula 1]

The pharmaceutical composition according to the present invention can provide an effect of treating or preventing prostate cancer, pancreatic cancer, colon cancer, lung cancer, head and neck cancer, esophageal cancer, gastrointestinal cancer, breast cancer, or kidney cancer.

The composition according to the present invention inhibits the activity of the calcium-dependent chloride ion channel (CaCC), and particularly treats or prevents cancer by inhibiting the activity of ANO1.

In addition, the composition according to the present invention may induce apoptosis in cancer cells.

In another embodiment of the present invention, the composition according to the present invention may be administered in combination with other anticancer agents.

Other anticancer agents as described above include, but are not limited to, cyclophosphamide (CTX), taxane (paclitaxel or docetaxel), cyclodextrin, cisplatin, cyclophosphamide, methotrexate, daunorubicin, doxorubicin, melphalan, cladribine, , Bin blastin or chlorambucil.

The present invention relates to a composition capable of treating or preventing cancer by inhibiting CaCC / ANO1 activity, wherein CaCC / ANO1 is overexpressed in 80% of head and neck squamous cell carcinoma, Prostate cancer, and cancer cells from respiratory or digestive epithelial cells. Therefore, the pharmaceutical composition comprising idenone according to the present invention has sufficient marketability as an anticancer agent. In addition, since currently used anticancer drugs do not have a drug targeting CaCC / ANO1, it has been developed as a new class of anticancer drugs and can be used alone or in combination with existing anticancer drugs.

1 is a graph showing the effect of inhibiting the activity of ANO1 in the presence of calcium signal by ibebenone. (a) is a graph showing the ANO1 inhibitory effect of FRP-ANO1 cells treated with 1, 3, 10, 30, and 100 μM of IFEn and YFP type and measurement, and (b) (Calcium-specific fluorescence) fluorescence after treatment with 30 μM ibebenone, and the effect of ibebenone on the intracellular calcium concentration was measured.
2 is a graph showing the effect of inhibiting the activity of CaCC by idebanone in HT-29 cells. (a) is a graph showing the CaCC / ANO1 inhibitory effect measured by fluorescence measurement of YFP after treating 1, 3, 10, 30, and 100 μM of idebenone in HT-29 cells, FIG. 4 is a graph showing the CaCC / ANO1 inhibitory effect measured by YFP fluorescence measurement after treating ibebenone with various concentrations in the cells and HT-29 cells.
FIG. 3 is a graph showing the inhibitory effect of idebenone on CaCC activity in FRT-ANO1, T-84 and Calu-3 cells through short-circuit current measurement. (B), T-84 (human colon cancer cell line) cells, (c) Calu-3 cells, and (Human lung cancer cell line) cells and observed CaCC / ANO1 inhibitory effect.
FIG. 4 is a graph showing the inhibitory effect of idebenone on CaCC / ANO1 activity through whole-cell patch clamp in FRT-ANO1 and PC3 cells. (A, D) is the result of the whole-cell CaCC current measurement, (B, E) is the current density graph, and (C, F) is the current / voltage (I / V) plot.
FIG. 5 shows the expression of ANO1 protein and the inhibitory effect of idebenone on the activity of CaCC / ANO1 in PC3 and CFPAC-1 cells in FRT-ANO1, PC3, CFPAC-1, HT-29, T-84, Calu- (A) and CFPAC-1 cells (B), respectively, showing changes in CaCC / ANO1 activity by YFP fluorescence measurement.
FIG. 6 is a graph showing the inhibitory effect of idebenone on cell growth in PC3, CFPAC-1, HT-29, T-84, Calu-3 and A549 cells. (A) is the MTS assay, and (B) is the BrdU assay.
FIG. 7 is a fluorescence image showing apoptosis induction effect by idebenone in PC3, CFPAC-1, HT-29, T-84, Calu-3 and A549 cells.

In one aspect, the invention provides a pharmaceutical composition for treating or preventing cancer comprising an effective amount of idebenone.

The above-mentioned ideone is represented by the following formula 1:

[Chemical Formula 1]

The present invention provides a method for treating or preventing cancer by specifically inhibiting the activity of calcium-dependent chloride ion channel (CaCC) and the activity of ANO1, which is one of CaCCs. Specifically, this is because ibebenone inhibits the activity of CaCC / ANO1 in various cancer cells expressing CaCC / ANO1 without affecting intracellular calcium signal, thereby causing inhibition of cancer cell growth and apoptosis. In accordance with the present invention, the inhibition of cell growth and the induction of apoptosis-inducing effect by ibebenone in various cancer cells were confirmed in Examples 6 and 7. Specifically, ibebenone is a prodrug of PC3 cells (prostate cancer cells), CFPAC-1 cells (pancreatic cancer cells), HT-29 and T-84 cells (colorectal cancer cells) and Calu-3 cells (lung cancer cells) expressing CaCC / ANO1 The effect of ibebenone on the growth of A549 cells (lung cancer cells), which do not express CaCC / ANO1, was found to be influenced by concentration of ibebenone. Respectively. It also revealed apoptosis in PC3, CFPAC-1, HT-29, T-84 and Calu-3 cells. Accordingly, the pharmaceutical composition containing ibebenone according to the present invention inhibits the growth of cancer cells expressing CaCC / ANO1 and induces apoptosis, thereby providing a pharmaceutical composition for treating or preventing cancer, which targets cancer expressing CaCC / ANO1 .

In the present invention, " cancer " is a disease caused by a variety of causes, such as abnormal cell growth, abnormal cell growth, abnormal invasion of surrounding tissues and organs, To form masses (masses) and to destroy or deform existing structures.

The composition according to the present invention can treat or prevent cancer. Here, the target cancer is characterized by expression of CaCC / ANO1, which may be prostate cancer, pancreatic cancer, colon cancer, lung cancer head and neck cancer, esophageal cancer, gastrointestinal cancer, breast cancer, or kidney cancer.

In the present invention, "prevention" means any action that inhibits or slows the onset of cancer by administration of the composition, and "treatment" It means all acts.

The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

In another embodiment of the present invention, the composition according to the present invention can be administered in combination with other anticancer drugs.

Here, other anticancer agents include cyclophosphamide (CTX), taxane (papillotaxel or docetaxel), biphenylyl, cisplatin, cyclophosphamide, methotrexate, dovorubicin, doxorubicin, melphalan , Cladribine, vincristine, vinblastine or chlorambucil.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, topical administration, transdermal administration and the like.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on such factors as formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate, . A preferred dosage of the pharmaceutical composition of the present invention is 2,000 mg / day to 2,500 mg / day on an adult basis, more preferably about 2,250 mg / day

&Quot; Effective amount " of the present invention means a therapeutically effective amount, which means that the amount of the drug is not less than the amount showing the action necessary for the treatment and less than the amount showing toxicity to the living body.

The pharmaceutical composition of the present invention may be formulated into a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Idebenone On by Calcium signal Independent ANO1 Activity inhibition

FRT (Fisher Rat Thyroid) cells expressing ANO1 / TMEM16A and YFP were cultured in a 96-well microplate for 48 hours with DMEM / Ham`s F12 1: 1 media. The culture medium was then removed and the cells were washed with a regular solution (140 mM NaCl, 5 mM KCl, 1 mM MgCl ₂ , 10 mM HEPES, 1 mM CaCl ₂ , 10 mM Glucose D (+), pH 7.4) were treated with Idebenone at concentrations of 0, 1, 3, 10 and 30 μM. After 10 minutes of reaction time of drug and cells, NaI solution (140 mM NaI, 5 mM KCl, 1 mM MgCl ₂ , 10 mM HEPES, 1 mM CaCl ₂ , 10 mM Glucose D (+), pH 7.4) containing 100 uM of ATP was added to each well, reader was used to measure the decrease in YFP fluorescence by iodide ions entering the cell through ANO1 at intervals of 0.4 seconds for 5 seconds. Excitation 488nm and emission 520nm were used for YFP fluorescence. All experiments were done at 37 < 0 > C (Figure 1A)

HT29 (human colon cancer cell line) cells are cultured in a 96-well RPMI1640 medium for 48 hours, then washed with PBS, and then 100 μl of Flou4 -NW dye mix is added and cultured for 1 hour in a dark state. After adding 10 μl of idebenone to the cells loaded with Fluo4-NW, the cells were reacted for 10 min. Then, a solution containing 100 μM of P2Y receptor activator ATP (100 μM) was added to the cells and the change of fluorescence was measured at intervals of 0.5 sec for 100 sec. Change was measured. Excitation 488nm and emission 520nm were used for the measurement fluorescence wavelength. All experiments were performed at 37 ° C (FIG. 1B).

In the FRT cells expressing ANO1, idebenone inhibited the ANO1 channel by concentration. In addition, idebenone specifically inhibited ANO1 without affecting intracellular calcium signal.

HT -29 cells idebenone Endogenously expressed by CaCC Activity inhibition

HT-29 cells expressing iodine-specific YFP are cultured in a 96-well plate for 48 hours in RPMI1640 medium, and then the culture medium is removed. Regular solution (100 μl) was added to each well of a 96-well plate. Idebenone was treated with concentration (1, 3, 10, 30, 100 μM) and reacted for 10 min. After that, 100 μl of ATP (100 μM) was added to the NaI solution, and the decrease of YFP fluorescence was measured, and the effect of CaCl 2 inhibition by idebenone was measured at intervals of 0.4 second for a total of 25 seconds (FIG. 2A).

Idebenone was treated at different concentrations (1, 3, 10, 30, 100 μM) in FRT cells and HT29 cells to determine the decrease in YFP fluorescence and to determine the dose response curve (FIG. 2B).

We confirmed that Idebenone inhibits CaCC / ANO1 by concentration in human colon cancer cell HT-29 cells.

Short - circuit current  Through measurement FTR - ANO1 , T-84 and Calu -3 cells by idebenone CaCC Activity inhibition

FRT cells expressing ANO1 / TMEM16A were cultured in Transwell for 5 days and differentiated. Short-circuit currents were measured using a Ussing chamber. There basolateral bath of the Ussing chamber RegularHCO ₃ ^- into the solution (120 mMNaCl, 5 mM KCl , 1 mM MgCl 2, 2.5 mM HEPES, 1mM CaCl 2, 10 mM Glucose, 25 mM NaHCO 3) luminal bath contains 70mM Cl ^- Put Cl ^- gradient was prepared and stabilized by adding 5% CO ₂ for 20 minutes. After that, idebenone was treated with luminal bath at concentration (3, 10, 30, 100 μM) for 10 min and ATP (100 μM) was added to measure the inhibitory effect of Cl ^- secretion through CaCC activity (see FIG. 3A).

T-84 (human colorectal cell line) cells expressing endogenous CaCC were cultured in transwell for 7 days in DMEM / Ham`s F12 1: 1 culture medium and short-circuit current was measured. After stabilization in a regulated HCO ₃ ^- solution for 20 min, 10 μM of CFTR _Inh -172 was added to the luminal bath to inhibit CFTR. Then, the CaCC current was increased by adding 100 μM of ATP, followed by treatment with idebenone (concentration of 1, 3, 10, 30, 100 μM) to observe CaCC inhibitory effect (see FIG.

Calu-3 (human lung carcinoma cell) cells expressing endogenous CaCC were cultured in transwell for 10 days in 1: 1 DMEM / Ham`s F12 culture medium, and short-circuit current was measured. After stabilization in regular HCO ₃ ^- solution for 20 min, 10 μM of CFTR _Inh -172 was added to inhibit the activity of CFTR. After 10 min of incubation with Idebenone (10, 30, 100 μM), ATP 100 μM was added to the basolateral bath And CaCC was activated to observe the effect of idebenone on CaCC (see FIG. 3C).

The short-circuit current measurement of idebenone inhibited CaCC / ANO1 activity in FRT cells expressing CaCC / ANO1, T-84, and Calu-3.

FRT - ANO1 Wow PC3  In a cell whole - cell patch clamp  Through practice idebenone Inhibition of CaCC / ANO1 activity by

Whole-cell patch clamps were performed in (AC) FRT-ANO1 cells expressing ANO1 and (DF) PC3 cells. The Pipette solution consisted of 140 mM NMDG-Cl, 0.5 mM EGTA, 1 mM MgCl ₂ , 1 mM Tris-ATP, and 10 mM HEPES, and the pH was adjusted to 7.2. The bath solution consisted of 140 mM NMDG-Cl, 1 mM CaCl2, 1 mM MgCl2, 10 mM glucose, and 10 mM HEPES, and the pH was adjusted to 7.4. All experiments were conducted at 22-25 ° C and the pipette resistance was 3-5 MΩ. CaCC / ANO1 was activated with 100 μM ATP and treated with 10 or 30 μM Idebenone to inhibit Cl ^- current. The whole-cell current was measured from +80 mV to -80 mV in 20 mV steps and the holding potential was 0 mV. (A, D) Whole-cell CaCC current measurement. (B, E) Current density graph. (C, F) Current / voltage (I / V) plot.

Using a patch clamp, 10 μM Idebenone inhibited ANO1 ion channel activity by about 50% in one FRT cell and prostate cancer cell (PC3) expressing high levels of ANO1. In PC3 cells, 30 μM Idebenone inhibited the ANO1 ion channel activity by about 90%.

FRT - ANO1 , PC3 , CFPAC -One, HT -29, T-84, Calu -3, in A549 cells ANO1  Protein expression and PC3 Wow CFPAC -1 cells idebenone On by CaCC / ANO1 Activity inhibition

PC3, CFPAC-1, HT-29, T-84, Calu-3, A549 and FRT-ANO1 cells were resuspended in lysis buffer (20 mM Tris-HCl pH 7.9, 137 mM NaCl, 5 mM EDTA, 10% Glycerol, 1 mM Na3VO4, 1 mM β-glycerolphosphate), and the resulting mixture was centrifuged at 13,000 rpm at 4 ° C. for 10 minutes to obtain a supernatant And the proteins were separated. To each protein sample, 5X sample buffer (313 mM Tris-HCl (pH 6.8), 208 mM dithiothreitol, 50% glycerol, 10% SDS and 0.05% BPB) was added and boiled at 60 ° C for 5 minutes. , KOMA BIOTECH, KOREA) and transferred to a polyvinylidene fluoride membrane (Millipore, Billerica, MA, USA). Membrane was blocked with 5% skim milk (Tris-buffered saline (TBS-T) containing 0.1% Tween 20) and incubated with primary antibody (anti-TMEM16A 1: 1,500, anti-actin 1: . Membrane was washed with TBS-T, incubated with secondary antibody conjugated with HRP for 1 hour at room temperature, washed with TBS-T, and then treated with enhanced chemiluminescent HRP substrate (ECL substrate, Clarity Western ECL Substrate, BIO-RAD, USA ) And membrane were reacted and developed, and the TMEM16A band was observed (Figure 5A)

CFPAC-1 cells were cultured in a 24-well plate for 24 hours and then transfected with YFP using lipofectamine. After 6 hours, the cells were replaced with fresh medium. After culturing for 24 hours, the medium was replaced with 200 μl of a regular solution. Tannic acid 100 μM and Idebenone 30 μM were added for 10 minutes. Then, 200 μl of NaI solution containing 100 μM ATP And measured with a microscope every 0.5 second (Fig. 5B).

ANO1, PC3, CFPAC-1, HT-29, T-84 and Calu-3 cells expressed ANO1 while A549 cells expressed ANO1 .

In the case of CFPAC-1 cells treated with Idebenone 30 μM, the inhibition of the ion channel activity of ANO1 by 95% or more was observed similar to that of tannic acid 100 μM.

PC3 , CFPAC -One, HT -29, T-84, Calu -3, in A549 cells idebenone To inhibit cell growth

(3, 10, and 30 μM) when cells were 30% confluent after culturing for 24 hours in a 96-well microplate. , 100 μM of Coenzyme Q10, 10 μM of T16A _ihn- A01, and 100 μM of tannic acid. The culture medium and drug were replaced every 12 hours. After 48 hours, 20 μl of MTS solution was added to each well, incubated at 37 ° C and 5% CO ₂ for 1 hour, and absorbance at 490 nm was measured with a plate reader. (Figure 6A)

PC3, CFPAC-1, Calu-3, A549, HT-29 and T-84 were cultured in 96-well microplates for 24 hours. After 30% of the cells were filled, Idebenone 30 μM, Coenzyme Q10 100 μM, T16A _ihn -A01 10 μM, tannic acid (100 μM). The culture medium and drug were changed every 12 hours. After 48 hours, 10 μM of BrdU labeling solution was added to each well and incubated for 4 hours at 37 ° C and 5% CO ₂ . After removing the culture solution, Fixdenat solution was added to each well at 200 ul each and reacted at room temperature for 30 minutes. Fixdenat solution was removed and 100 μl of anti-BrdU solution was added. The reaction was allowed to proceed at room temperature for 90 minutes. Thereafter, the cells were washed 3 times with washing solution for 5 minutes. Then, 100 μl of TMB substrate was added to each well. After incubation for 25 minutes at room temperature, the absorbance was measured at 370 nm / 492 nm (FIG.

MTS assay and BrdU assay showed that 30 μM of idebenone inhibited cell growth of CaCC / ANO1-expressing cells (PC3, CFPAC-1, Calu-3, HT-29 and T-84) But not cell growth of A549 cells.

PC3 , CFPAC -One, HT -29, T-84, Calu -3, in A549 cells idebenone On by apoptosis  Identification of inducing effect

PC3, HT-29, CFPAC-1, A549, Calu-3 and T-84 cells were cultured on a 96-well black plate for 24 hours. Idebenone 30 μM was then treated for 48 hours. After incubation, the cells were fixed with 1% paraformaldehyde, ehanol, acetic acid, and TdTenzyme was added. The following anti-digoxigenin conjugate was added and reacted at room temperature for 30 minutes, and then stained with DAPI and mounted. Each step was washed three times with 200 μl of PBS. Cells with apoptosis were observed using a fluorescence microscope (Fig. 7).

Apoptosis (green fluorescence) occurred in PC3, HT-29, CFPAC-1, A549, Calu-3 and T-84 cells treated with Idebenone 30 μM for 48 hours and apoptosis occurred in A549 cells not expressing ANO1 .

Claims (10)

1. A composition for treating or preventing cancer comprising an ibebenone compound represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

The pharmaceutical composition according to claim 1, wherein the cancer is selected from the group consisting of prostate cancer, pancreatic cancer, colon cancer, lung cancer, head and neck cancer, esophageal cancer, gastrointestinal cancer, breast cancer and kidney cancer.
2. The pharmaceutical composition according to claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier.
The pharmaceutical composition according to claim 1, wherein the composition inhibits the activity of the calcium-dependent chloride ion channel (CaCC).
5. The pharmaceutical composition according to claim 4, wherein the calcium-dependent chloride ion channel is ANO1.
2. The pharmaceutical composition according to claim 1, wherein said composition induces apoptosis in cancer cells.
The pharmaceutical composition according to claim 1, wherein said composition can be used in combination with other anticancer agents.
8. The pharmaceutical composition according to claim 7, wherein the anticancer agent is selected from the group consisting of cyclophosphamide (CTX), taxane (papillotaxel or docetaxel), vesicular, cisplatin, cyclophosphamide, methotrexate, daunorubicin, doxorubicin, melphalan, cladribine , Vincristine, vinblastine and chlorambucil. ≪ RTI ID = 0.0 > 8. < / RTI >
4. The composition of claim 3 wherein the pharmaceutically acceptable carrier is selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone , At least one selected from the group consisting of cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
The pharmaceutical composition according to claim 1, wherein the dosage of the pharmaceutical composition is 2,000 mg / day to 2,500 mg / day.

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