KR102489977B1 - Pharmaceutical composition for the prevention or treatment of cancer, comprising isosakuranetin or pharmaceutically acceptable salts thereof as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer comprising Isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient, including colorectal cancer (HCT116), lung cancer (A549), breast cancer (MDA) -MB-231) As it was confirmed that treatment of cells with Isosakuranetin stopped the cell proliferation and cell cycle of cancer cells and induced apoptosis and autophagy, A composition containing Isosakuranetin to inhibit growth, recurrence, and metastasis and to kill cancer cells is provided as an anti-cancer agent.

Description

Pharmaceutical composition for the prevention or treatment of cancer, comprising isosakuranetin or pharmaceutically acceptable salts thereof as an active ingredient}

The present invention relates to a pharmaceutical composition for preventing or treating cancer comprising isosacuranetin or a pharmaceutically acceptable salt thereof as an active ingredient.

Cancer is a disease caused by an increase in the number of abnormal cells derived from normal tissues, and enormous capital is being invested in development to cure cancer worldwide. Cancer is known to be caused by various carcinogen factors such as smoking, ultraviolet rays, chemicals, food, and other environmental factors. However, in the past, various studies have been conducted to develop suitable therapeutic agents according to the cause of onset, but in the case of the developed therapeutic agent, it has been reported that the therapeutic effect is different depending on the cause of onset or onset site.

Cancer malignancy is a major cause of death of patients in large hospitals, and since more than half of the world's deaths are due to cancer malignancy, many studies are being conducted on cancer treatments or treatments that inhibit cancer malignancy. In the treatment of cancer, surgery, radiation therapy, development of anticancer drugs, and biological methods have been developed. Research has been conducted to develop anticancer drugs using various types of cancer cells.

However, since some cancer therapeutics are toxic at high concentrations, it is difficult to selectively remove only cancer cells, which causes various side effects. there is. Therefore, it is urgently needed to develop an anticancer agent with less toxicity that can prevent malignant transformation of cancer as well as exhibit cancer prevention and treatment effects.

On the other hand, as it is known statistically that the risk of cancer is reduced when a large amount of some fruits or vegetables are consumed, research is being conducted to find components exhibiting anticancer effects in natural resources. In particular, as it has been reported that antioxidant, anti-inflammatory, anti-cancer, and anti-viral effects appear in flavonoid compounds abundant in tea, fruits and vegetables, various flavonoid compounds are attracting attention as candidates for anti-cancer drugs.

Korean Patent Publication No. 10-2018-0090122

Chemical biology & drug design 89.3 (2017): 353-364.

The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer comprising Isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient, including colorectal cancer (HCT116), lung cancer (A549), breast cancer (MDA) -MB-231) As it was confirmed that treatment of cells with Isosakuranetin stopped the cell proliferation and cell cycle of cancer cells and induced apoptosis and autophagy, To inhibit growth, recurrence, and metastasis, and to provide a composition containing Isosakuranetin to kill cancer cells as an anti-cancer agent.

The present invention provides a pharmaceutical composition for preventing or treating cancer comprising Isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving cancer comprising Isosakuranetin or a salt acceptable as a food ingredient thereof as an active ingredient.

Or Isosakuranetin (Isosakuranetin) or a pharmaceutical composition for inhibiting recurrence or metastasis of cancer containing a pharmaceutically acceptable salt thereof as an active ingredient.

According to the present invention, by treating colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells with Isosakuranetin, the cell proliferation and cell cycle of cancer cells are stopped and apoptosis occurs. As it is confirmed that apoptosis and autophagy are induced, a composition containing Isosakuranetin to suppress cancer growth, recurrence, and metastasis, and kill cancer cells as an anticancer agent. can be provided.

1 is a graph evaluating the cell proliferation inhibitory ability of Isosakuranetin according to its concentration in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
FIG. 2 is a photograph confirming the ability of Isosakuranetin to inhibit wound healing in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
Figure 3 is a graph evaluating the wound healing inhibition ability after 24 hours of isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells. The wound percentage (wound %) was evaluated on the basis of 100% at 0 time zone before treatment.
4 is a graph showing histograms of each cell cycle according to Isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
5 is a graph showing the percentage (%) of cells occupying each cell cycle according to Isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
FIG. 6 is a photograph showing changes in the expression of G1 cell cycle-related proteins according to Isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
FIG. 7 is a photograph showing changes in expression of apoptosis-related proteins according to Isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
8 is a photograph confirming the change in autophagy according to Isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells using a fluorescence microscope.
FIG. 9 is a photograph showing changes in the expression of autophagy-related proteins according to Isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
FIG. 10 is a photograph showing changes in the expression of proteins related to the AKT-mTOR pathway according to Isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.

The terms used in this specification have been selected from general terms that are currently widely used as much as possible while considering the functions in the present invention, but these may vary depending on the intention of a person skilled in the art, precedent, or the emergence of new technologies. In addition, in a specific case, there is also a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the invention. Therefore, the term used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, not simply the name of the term.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and unless explicitly defined in this application, it should not be interpreted in an ideal or excessively formal meaning. don't

Numerical ranges are inclusive of the values defined therein. Every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written. Every minimum numerical limitation given throughout this specification includes every higher numerical limitation, as if such higher numerical limitations were expressly written. Every numerical limitation given throughout this specification will include every better numerical range within the broader numerical range, as if the narrower numerical limitations were expressly written.

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

The present inventors have completed the present invention by confirming that Isosakuranetin inhibits the growth of cancer cells and exhibits an effect of inducing death of cancer cells in view of this point.

The present invention provides a pharmaceutical composition for preventing or treating cancer comprising Isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient.

Isosakuranetin is a compound represented by Formula 1 below and is a kind of flavanone.

[Formula 1]

Isosakuranetin has a molecular formula of C ₁₆ H ₁₄ O ₅ and a molecular weight of 286.3, and is 4'-methylnaringenin or (2S)-5,7-dihydroxy-2-(4). Also called -methoxyphenyl)-2,3-dihydrochromen-4-one ((2S)-5,7-dihydroxy-2-(4-methoxyphenyl)-2,3-dihydrochromen-4-one) It became. Isosakuranetin is also known as an aglycone compound of poncirin, a component of Poncirus trifoliata , and can be extracted with solvents such as chloroform, dichloromethane, ethyl acetate, DMSO, and acetone. can

The cancer may be colorectal cancer, lung cancer, breast cancer, stomach cancer, liver cancer, pancreatic cancer, uterine cancer, esophageal cancer, bladder cancer, head and neck cancer, leukemia, lymphoma, hematological cancer, rectal cancer, prostate cancer, ovarian cancer, kidney cancer, brain cancer, or skin cancer. .

The isosakuranetin inhibits the proliferation of cancer cells and stops the cell cycle at the G1 phase, thereby inhibiting cancer growth, recurrence, and metastasis. In addition, the isosakuranetin has an activity of killing cancer cells by inducing apoptosis and autophagy of cancer cells.

The pharmaceutical composition may further include carriers, excipients or diluents commonly used in the preparation of pharmaceutical compositions.

The pharmaceutical composition may be formulated in the form of one or more external agents selected from the group consisting of creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma agents.

The pharmaceutical composition of the present invention may further contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose, gelatin, alginates, and polyvinyl fibres. Binders such as rolidone, etc., lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone, crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol. It doesn't work. The pharmaceutically acceptable carrier and diluent may be biologically and physiologically compatible with the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents and/or dispersion media.

The pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the desired method, and is preferably administered orally. In addition, the dosage of the pharmaceutical composition of the present invention varies depending on the subject's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease, but is not limited thereto.

In addition, the present invention provides a health functional food composition for preventing or improving cancer comprising Isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention can be generally used as a commonly used food.

The food composition of the present invention can be used as a health functional food. The term "health functional food" refers to food manufactured and processed using raw materials or ingredients having useful functionalities for the human body in accordance with the Health Functional Food Act, and "functional" refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.

The food composition of the present invention may include conventional food additives, and the suitability as the "food additive" is determined according to the general rules of the food additive code approved by the Ministry of Food and Drug Safety and general test methods, etc., unless otherwise specified. It is judged according to the specifications and standards for the item.

Items listed in the "Food Additive Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.

The food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like.

For example, among the health functional foods in the form of capsules, hard capsules can be prepared by mixing and filling the composition according to the present invention with additives such as excipients in conventional hard capsules, and soft capsules can be prepared by filling the composition according to the present invention. It can be prepared by mixing with additives such as excipients and filling in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

Definitions of terms for the excipients, binders, disintegrants, lubricants, corrigents, flavoring agents, etc. are described in literature known in the art, and include those having the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in the usual sense.

In the present invention, the term "prevention" refers to all activities that inhibit or delay a disease by administering the composition according to the present invention. In the present invention, the term "treatment" refers to all activities that improve or beneficially change the symptoms of a disease by administering the composition according to the present invention. In the present invention, "improvement" means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to a subject.

Hereinafter, experimental examples and examples will be described in detail to aid understanding of the present invention. However, the following experimental examples and examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following experimental examples and examples. Experimental examples and examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

<Experimental Example> Experimental Materials and Methods

The following experimental examples are intended to provide experimental examples commonly applied to each embodiment according to the present invention.

1. Cell culture and materials

Colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells used in this experiment were purchased from the Korean Cell Line Bank. Each cell was cultured in a 5% CO ₂ incubator at 37° C. by adding 10% FBS and 1% penicillin to DMEM medium (Wellgene).

Isosakuranetin used in this experiment was directly synthesized by the inventor with reference to a known manufacturing method (Hong-shun Gu1 et al. 2017).

2. Cell proliferation evaluation

Colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were counted and inoculated in 3 X 10 ⁴ ~ 5 X 10 ⁴ each in 96-well plates, and then inoculated for 12 hours. It was cultured in a DMEM medium (Wellgene) containing 1% FBS and 1% penicillin in a 5% CO ₂ incubator. The cultured cells were treated with Isosakuranetin at a concentration of 0 μM to 700 μM, and cultured in an incubator at 37° C. and 5% CO ₂ for 72 hours. After the culture was completed, to evaluate the degree of cell proliferation inhibition, the medium was removed and 0.5% MTT solution (3-[4,5-Dimethylthiazol-2yl]-2,5-diphenyl-etrazolium Bromide) (Sigma-Aldrich Korea) was added. It was mixed with serum-free medium and treated at a ratio of 1/10, and after 2 hours, the number of cells was measured by 590 nm absorbance.

3. Evaluation of wound healing

Colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were inoculated into 6-well dishes, respectively, and inoculated with 10% FBS and 1% penicillin until the inoculated cells covered the entire dish. DMEM medium (Wellgene) containing this was cultured in an incubator at 37°C and 5% CO ₂ . The dish in which the cells were cultured was treated with 25 μg/ml of mitomycin C (Sejinshiai) for 30 minutes, and an injury line was made in the center of the dish with a 200 mL tip. After treating each dish with 300 μM of Isosakuranetin, the degree of cell filling in the injury line was observed for 24 hours at each time point. The degree of wound healing was evaluated by the change in the width of the injury line over time based on the width of the injury line at time 0 as 100%.

4. Flow cytometry

Colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were inoculated into 60 mm plates, respectively, and DMEM medium (Wellgene) containing 10% FBS and 1% penicillin for 12 hours. 37 ℃, 5% CO ₂ It was cultured in an incubator. It was treated with 300 μM of Isosakuranetin and cultured for 24 hours. After recovering the cultured cells and fixing the cell cycle with 70% ethanol, 50 μg/ml propidium iodine (Abcam) and RNase A (Qiagen) were treated for 30 minutes. The cell cycle of each cancer cell was observed using a flow cytometer (BD FACSDiva 7.0 software program).

5. Immunofluorescence flow cytometry

Colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were inoculated into a 24-well plate with a 12-mm coverslip, respectively, and treated with 10% FBS and 1% penicillin for 12 hours. It was cultured in a 37°C, 5% CO ₂ incubator with DMEM medium (Wellgene). After treatment with 300 μM of Isosakuranetin and incubation for 24 hours, the cell cycle was fixed by treatment with 4% paraformaldehyde for 20 minutes. After washing each cell with PBS (phosphate buffer saline) (RD tech), it was treated with 0.1% Triton-X 100 for 20 minutes. LC3B (light chain 3 antibody), an autophagy marker, was treated at a ratio of 1:200 and incubated at 4°C for 12 hours. Then, after treatment with Alexa Fluor 488 secondary antibody (Invitrogen) for 1 hour at room temperature, DAPI (4′,6-diamidino-2-phenylindol) (Sigma-Aldrich) for 5 minutes and confocal microscopy A fluorescence image of each cell was confirmed.

6. Immunoblot Analysis

Colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were inoculated into 60 mm plates, respectively, and DMEM medium (Wellgene) containing 10% FBS and 1% penicillin for 12 hours. 37 ℃, 5% CO ₂ It was cultured in an incubator. It was treated with 300 μM of Isosakuranetin and cultured for 24 hours. RIPA lysis buffer (Ato Technology) containing a protease inhibitor cocktail was treated for 30 minutes, and each cell was centrifuged to obtain proteins in the supernatant. After measuring the concentration of the recovered proteins with the BCA kit (Bicinchonic Acid kit), 30 μg of each protein was subjected to electrophoresis on a 7.5-10% SDS-PAGE gel. The protein separated on the gel was transferred to a nitrocellulose (NC) membrane, and the NC membrane was treated with 5% skim milk for 1 hour. The primary antibody diluted with 5% BSA (Bovine Serum Albumin) was treated at a ratio of 1:1,000 and incubated at 4°C for 12 hours. Thereafter, the secondary antibody diluted with 5% skim milk was treated at a ratio of 1:10,000 at room temperature for 1 hour. Each protein band was identified using ImageQuant LAS-4000 mini (GE Healthcare Life Sciences) using ECL solution (Thermo Fisher Scientific).

7. Statistics

All experiments were repeated three times and expressed as mean±standard deviation (SD). On each data, * means P < 0.05, ** means P < 0.01.

Example 1. Inhibition of cell proliferation of cancer cells according to Isosakuranetin treatment

In order to evaluate the proliferation change of cancer cells according to Isosakuranetin treatment, Isosakuranetin was administered to colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells. After treatment, the number of cells was measured.

As shown in Figure 1, colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells are inhibited in cell proliferation in proportion to the concentration of Isosakuranetin. As shown in Table 1 below, IC50, which is a concentration at which the growth of colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells are inhibited by 50%, was 349.7 and 114.4, respectively. and 594.8 μM.

The above results confirmed that Isosakuranetin has an effect of inhibiting the proliferation of colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, which is Isosakuranetin) has an anticancer effect, and it proves that it has an activity to inhibit cancer growth, recurrence, and metastasis.

Example 2. Inhibition of cancer cell wound healing by isosakuranetin treatment

In order to evaluate changes in the wound healing ability of cancer cells according to Isosakuranetin treatment, isosakuranetin (Isosakuranetin ) was treated and the change in the degree of wound healing was measured.

As shown in Figures 2 and 3, compared to the control group in which isosakuranetin was not treated in colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, isosakuranetin ( Isosakuranetin), it was found that the degree of wound healing after 24 hours was inhibited.

The above results confirmed that Isosakuranetin has an effect of inhibiting wound healing of colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, which is (Isosakuranetin) has an anticancer effect, and it is proved that there is an activity to inhibit cancer growth, recurrence and metastasis.

Example 3. Cell cycle change of cancer cells according to isosakuranetin treatment

To evaluate the cell cycle of cancer cells following treatment with Isosakuranetin, colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were treated with Isosakuranetin. After treatment, flow cytometry was performed, and the expression of proteins related to the cell cycle was measured.

As shown in Figures 4 and 5, the cell cycle is arrested in G1 phase by treating colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells with Isosakuranetin. As a result, cells in S phase and G2/M phase were reduced. In addition, when Isosakuranetin was treated, the G1 cell number of each cancer cell increased compared to the control group. In cells, the number of G1-phase cells increased by 11.8% from 73.8% to 82.5%, and in breast cancer (MDA-MB-231) cells, the number of G1-phase cells increased by 25.6% from 63.3% to 79.5%.

In addition, as shown in FIG. 6, by treating colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells with Isosakuranetin, p53, known as a tumor suppressor, expression was found to increase. In addition, the expression of p21, a cyclin-dependent kinase inhibitor or CDK-interacting protein 1, was increased, and the expression of p15, which is known to inhibit the cyclin D1 and CDK4 complex, was increased in colorectal cancer cells. Expressions of Cyclin D1 and CDK4 involved in the G1 phase were all decreased, and expression of pRB was also decreased. As a result, the expression of Cyclin E, Cyclin A, and CDK2 involved in S phase was reduced.

The above results confirmed that Isosakuranetin has an effect of inhibiting the cell cycle of colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, which is (Isosakuranetin) has an anticancer effect, and it is proved that there is an activity to inhibit cancer growth, recurrence and metastasis.

Example 4. Induction of apoptosis of cancer cells according to Isosakuranetin treatment

In order to evaluate the apoptotic changes of cancer cells according to isosakuranetin treatment, colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were treated with Isosakuranetin ) and the expression level of apoptosis-related proteins was measured.

As shown in Figure 7, as isosakuranetin is treated in colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, the cleavage of PARP, a marker of apoptosis, It was observed that pro-apoptotic factors such as Bim increased, and the expression of Bcl-2, an anti-apoptotic marker, decreased.

The above results confirmed that Isosakuranetin has an effect of inducing apoptosis of colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, which is (Isosakuranetin) has an anticancer effect, and it proves that there is an activity to kill cancer cells.

Example 5. Induction of autophagy of cancer cells according to Isosakuranetin treatment

In order to evaluate changes in autophagy in cancer cells following Isosakuranetin treatment, colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were treated with Isosakuranetin ), and changes in autophagy were observed under a fluorescence microscope, and the expression level of apoptosis-related proteins was measured. In order to compare the autophagy-inducing ability of Isosakuranetin, autophagy inhibitors chlorquin (CQ) or 3-MA were additionally treated.

As shown in FIG. 8, it was found that autophagy was induced under a fluorescence microscope by treating colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells with Isosakuranetin. appear. In addition, as shown in Figure 9, as isosakuranetin is treated in colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, the autophagy-related marker LC3B expression has been shown to be induced.

The above results confirmed that Isosakuranetin has an effect of inducing autophagy in colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, which is (Isosakuranetin) has an anticancer effect, and it proves that there is an activity to kill cancer cells.

Example 6. Changes in the AKT-mTOR pathway of cancer cells according to Isosakuranetin treatment

To evaluate changes in the AKT-mTOR pathway in cancer cells following Isosakuranetin treatment, isosakuranetin was administered to colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells. (Isosakuranetin), and the expression level of proteins related to the AKT-mTOR pathway was measured.

As shown in FIG. 10, p-Erk, p-AKT (Ser473 ), p-mTOR (Ser2448), p-P70S6K (Thr421/Ser424), and p-S6RP expressions were reduced.

The above results demonstrate that the cell proliferation inhibitory activity of Isosakuranetin is related to the AKT-mTOR pathway.

Having described specific parts of the present invention in detail above, it is clear to those skilled in the art that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. Do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (8)

A pharmaceutical composition for preventing or treating cancer comprising Isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient, wherein the Isosakuranetin inhibits the proliferation of cancer cells or A pharmaceutical composition characterized in that it has an activity to inhibit the growth of cancer by stopping the cell cycle in the G1 phase, wherein the cancer is selected from the group consisting of colorectal cancer, lung cancer and breast cancer. delete delete The pharmaceutical composition according to claim 1, wherein the isosakuranetin has an activity of killing cancer cells by inducing apoptosis or autophagy of cancer cells. A health functional food composition for preventing or improving cancer comprising Isosakuranetin or a salt acceptable as a food ingredient thereof as an active ingredient, wherein the cancer is selected from the group consisting of colorectal cancer, lung cancer and breast cancer Health functional food composition. The method of claim 1,
The pharmaceutical composition for preventing or treating cancer, characterized in that the pharmaceutical composition is for cancer patients in which p-AKT, p-mTOR or p-Erk proteins are overexpressed. A pharmaceutical composition for suppressing recurrence or metastasis of cancer comprising Isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient, wherein the Isosakuranetin inhibits the proliferation of cancer cells or A pharmaceutical composition characterized in that it has an activity to inhibit recurrence and metastasis of cancer by stopping the cell cycle in G1 phase, wherein the cancer is selected from the group consisting of colorectal cancer, lung cancer and breast cancer. The pharmaceutical composition for inhibiting recurrence or metastasis of cancer according to claim 7, which is for cancer patients in which p-AKT, p-mTOR or p-Erk proteins are overexpressed.

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