KR20210156462A - 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 preventing or treating cancer, containing, as an active ingredient, isosakuranetin or a pharmaceutically acceptable salt thereof. It has been identified that cancer cell proliferation and the cell cycle are arrested and apoptosis and autophagy are induced according to the treatment of colorectal cancer cells (HCT116), lung cancer cells (A549), and breast cancer cells (MDA-MB-231) with isosakuranetin, and thus provided, as an anticancer preparation, is a composition containing isosakuranetin in order to inhibit cancer growth, recurrence and metastasis and cause cancer cell apoptosis.

Description

A pharmaceutical composition for the prevention or treatment of cancer, comprising isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient, the 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 that is caused by an increase in the number of abnormal cells derived from normal tissues, and huge capital is invested in development to cure cancer worldwide. It is known that cancer is caused by various carcinogen factors such as smoking, ultraviolet rays, chemicals, food, and other environmental factors. However, in the prior art, various studies are being conducted to develop a suitable therapeutic agent according to the cause of the disease. However, in the case of the developed therapeutic agent, it has been reported that the therapeutic effect appears differently depending on the cause or the site of the disease.

Malignancy of cancer is the main cause of death of patients in large hospitals, and since more than half of the world's patients die due to malignancy of cancer, a lot of research is being done on therapeutic agents for cancer or therapies to suppress malignancy of cancer. In the treatment of cancer, surgery, radiation therapy, development of anticancer drugs, biological methods, etc. 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. have. Therefore, there is an urgent need to develop an anticancer agent with low toxicity that can prevent cancer from becoming malignant as well as exhibiting preventive and therapeutic effects.

On the other hand, as it is statistically known that the risk of cancer is reduced when a large amount of some fruits or vegetables is consumed, research is being conducted to find ingredients showing anticancer effects from natural sources. In particular, as it has been reported that the effects of antioxidant, anti-inflammatory, anticancer, antiviral, etc. appear in flavonoid compounds abundant in tea, fruits and vegetables, various flavonoid compounds are attracting attention as candidates for anticancer 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, colon cancer (HCT116), lung cancer (A549), breast cancer (MDA) -MB-231) cell proliferation and cell cycle of cancer cells are stopped as cells are treated with Isosakuranetin, and apoptosis and autophagy are induced. In order to inhibit growth, recurrence, and metastasis, and to kill cancer cells, a composition containing isosakuranetin is provided as an anticancer 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 the prevention or improvement of cancer comprising isosakuranetin or an acceptable salt thereof as a food ingredient as an active ingredient.

Or a pharmaceutical composition for inhibiting recurrence or metastasis of cancer comprising isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient.

According to the present invention, as colon cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells are treated with isosakuranetin, cell proliferation and cell cycle of cancer cells are stopped, and apoptosis As it was confirmed that (apoptosis) and autophagy were induced, a composition containing Isosakuranetin was used as an anticancer agent to inhibit cancer growth, recurrence, and metastasis, and to kill cancer cells. can be provided.

1 is a graph evaluating the cell proliferation inhibitory ability of isosakuranetin according to the concentration in colon cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
2 is a photograph confirming the wound healing inhibitory ability of isosakuranetin in colon cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
3 is a graph evaluating the wound healing inhibitory ability 24 hours after isosakuranetin treatment in colorectal cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells. Wound specific gravity (wound %) was evaluated based on 100% of time 0 before treatment.
4 is a graph showing a histogram 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.
6 is a photograph evaluating the expression change of G1 cell cycle-related protein according to isosakuranetin treatment in colon cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
7 is a photograph evaluating the expression change 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 changes in autophagy according to isosakuranetin treatment in colon cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells with a fluorescence microscope.
9 is a photograph evaluating the expression change of autophagy-related proteins according to isosakuranetin treatment in colon cancer (HCT116), lung cancer (A549), and breast cancer (MDA-MB-231) cells.
10 is a photograph evaluating the expression change of AKT-mTOR pathway-related protein 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 as currently widely used general terms as possible while considering the functions in the present invention, but these may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, and the like. In addition, in a specific case, there is a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the corresponding 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, rather than the name of a simple term.

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

Numerical ranges are inclusive of the values defined in that range. Every maximum numerical limitation given throughout this specification includes all lower numerical limitations as if the lower numerical limitation were expressly written. Every minimum numerical limitation given throughout this specification includes all higher numerical limitations as if the higher numerical limitation were expressly written. All numerical limitations given throughout this specification will include all numerical ranges that are better within the broader numerical limits, 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 induces apoptosis 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.

The isosakuranetin (Isosakuranetin) is a compound represented by the following formula (1), a kind of flavanone (flavanone).

[Formula 1]

The isosakuranetin (Isosakuranetin) has a molecular formula C ₁₆ H ₁₄ O ₅ , a molecular weight of 286.3, and 4'-methylnaringenin (4'-methylnaringenin) or (2S)-5,7-dihydroxy-2-(4 -Methoxyphenyl)-2,3-dihydrochromen-4-one ((2S)-5,7-dihydroxy-2-(4-methoxyphenyl)-2,3-dihydrochromen-4-one) 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, blood cancer, rectal cancer, prostate cancer, ovarian cancer, kidney cancer, brain cancer, or skin cancer .

The isosakuranetin has an activity of inhibiting cancer cell proliferation and arresting the cell cycle in the G1 phase, thereby inhibiting cancer growth, recurrence, and metastasis. In addition, the isosakuranetin (Isosakuranetin) has an activity to kill cancer cells by inducing apoptosis and autophagy of cancer cells.

The pharmaceutical composition may further include a carrier, excipient or diluent commonly used in the preparation of pharmaceutical compositions.

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

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier and diluent for formulation. The pharmaceutically acceptable carriers and diluents include starch, sugar, and excipients such as mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose, hydroxypropylcellulose, gelatin, alginate, and polyvinyl blood. binders such as rolidone, lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone and crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol, and the like. doesn't happen 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 (eg, intravenously, subcutaneously, intraperitoneally or topically) according to a 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 status, 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 product.

The food composition of the present invention can be used as a health functional food. The term "health functional food" means a food manufactured and processed using raw materials or ingredients useful for the human body in accordance with the Health Functional Food Act, and "functionality" refers to the structure and function of the human body. It refers to ingestion for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological effects.

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

The items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; Mixed preparations such as sodium L-glutamate preparation, noodle-added alkali agent, preservative agent, and tar color agent can be mentioned.

The food composition of the present invention may be manufactured 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 a conventional hard capsule with the composition according to the present invention with additives such as excipients, and the soft capsule is a composition according to the present invention. It can be manufactured by mixing with additives such as excipients and filling in capsule bases such as gelatin. The soft capsule formulation may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

The term definitions for the excipients, binders, disintegrants, lubricants, flavoring agents, and the like are described in documents known in the art and include those having the same or similar functions. The type of food is not particularly limited, and includes all health functional foods in the ordinary sense.

In the present invention, the term “prevention” refers to any act of inhibiting or delaying a disease by administering the composition according to the present invention. In the present invention, the term “treatment” refers to any action that improves or beneficially changes the symptoms of a disease by administration of the composition according to the present invention. In the present invention, "improvement" refers to any action that improves the bad state of a disease by administering or ingesting the composition of the present invention to an individual.

Hereinafter, experimental examples and examples will be described in detail to help the understanding of the present invention. However, the following experimental examples and examples are only to illustrate the content of the present invention, 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 of ordinary skill in the art.

<Experimental example> Experimental material and method

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

Colorectal 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 synthesized by the inventor with reference to a known preparation method (Hong-shun Gu1 et al. 2017).

2. Cell proliferation evaluation

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

3. Wound healing evaluation

Inoculate each 6-well dish with colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, and then add 10% FBS and 1% penicillin until the inoculated cells cover the dish. Incubated in an incubator at _{37 ° C., 5% CO 2} with DMEM medium (Wellgene) containing this. The cell cultured dish was treated with 25 μg/ml of mitomycin C (Sejinsiai) for 30 minutes, and a 200 mL tip was used in the middle of the dish to make an injury line. After treating each dish with 300 μM of Isosakuranetin, the degree of cell filling in the wound line was observed for 24 hours. The degree of wound recovery was evaluated by the change in the width of the wound line (injury line) over time based on 100% of the width of the wound line (injury line) at time 0.

4. Flowcytometry

Colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were inoculated on 60 mm plates, respectively, and DMEM medium containing 10% FBS and 1% penicillin for 12 hours (Wellgene) 37 ℃, 5% CO ₂ Incubated in an incubator. Isosakuranetin was treated with 300 μM and cultured for 24 hours. After recovering the cultured cells and fixing the cell cycle with 70% ethanol, 50 μg/ml propidium iodine (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. Immunoflowcytometry

Colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were each inoculated in a 24-well plate with a 12-mm coverslip, and 10% FBS and 1% penicillin were added for 12 hours. It was cultured in an incubator at 37° C., 5% CO _{2 with the included DMEM medium (Wellgene).} Isosakuranetin (Isosakuranetin) was treated with 300 μM, and after 24 hours of incubation, 4% paraformaldehyde (paraformaldehyde) was treated for 20 minutes to fix the cell cycle. After washing each cell with PBS (phosphate buffer saline) (RD tech), 0.1% Triton-X 100 was treated 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. After that, Alexa Fluor 488 secondary antibody (Invitrogen) was treated at room temperature for 1 hour, and then treated with DAPI (4′,6-diamidino-2-phenylindol) (Sigma-Aldrich) for 5 minutes, followed by confocal microscopy. A fluorescence image of each cell was confirmed.

6. Immunoblot Analysis

Colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were inoculated on 60 mm plates, respectively, and DMEM medium containing 10% FBS and 1% penicillin for 12 hours (Wellgene) 37 ℃, 5% CO ₂ Incubated in an incubator. Isosakuranetin was treated with 300 μM 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 a protein in the supernatant. After measuring the concentration of the recovered protein with the BCA kit (Bicinchonic Acid kit), 30 μg of each protein was electrophoresed on 7.5-10% SDS-PAGE gel. The protein separated on the gel was transferred to a NC (nitrocellulose) membrane, and the NC membrane was treated with 5% skim milk for 1 hour. 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 with ImageQuant LAS-4000 mini (GE Healthcare Life Sciences) using ECL solution (Thermo Fisher Scientific).

7. Statistics

All experiments were repeated 3 times and expressed as mean±standard deviation (SD). In each data, * means P < 0.05, ** means P < 0.01.

Example 1. Inhibition of cell proliferation of cancer cells by isosakuranetin treatment

Isosakuranetin was administered to colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells to evaluate changes in cancer cell proliferation following isosakuranetin treatment. treated and the number of cells was counted.

As shown in Figure 1, colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells in proportion to the concentration of processing isosakuranetin (Isosakuranetin) cell proliferation is inhibited. As shown in Table 1 below, the IC50, the concentration at which the growth of colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells was 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 proves that it has an activity to inhibit cancer growth, recurrence and metastasis.

Example 2. Inhibition of wound healing of cancer cells by treatment with Isosakuranetin

In order to evaluate the change in the wound healing ability of cancer cells following isosakuranetin treatment, isosakuranetin was applied to colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells. ) was treated and the change in the degree of wound healing was measured.

As shown in Figures 2 and 3, colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells compared to the control group not treated with isosakuranetin (Isosakuranetin) Isosakuranetin) was shown to inhibit wound healing after 24 hours.

The above results confirmed that isosakuranetin has the effect of inhibiting wound healing of colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells, which is isosakuranetin. (Isosakuranetin) has an anticancer effect and proves that it has 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 isosakuranetin treatment, isosakuranetin was administered to colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells. After processing, flow cytometry was performed, and the expression of proteins related to the cell cycle was measured.

As shown in Figures 4 and 5, colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells are treated with isosakuranetin, the cell cycle is stopped in the G1 phase It was found that, as a result, cells in S phase and G2/M phase were decreased. In addition, when isosakuranetin was treated, the number of G1 cells in each cancer cell increased compared to the control group, and in colon cancer (HCT116) cells, the number of G1 cells increased by 24.3% from 57.5% to 71.5%, and lung cancer (A549) In cells, the number of G1 cells increased by 11.8% from 73.8% to 82.5%, and in breast cancer (MDA-MB-231) cells, the number of G1 cells increased by 25.6% from 63.3% to 79.5%.

In addition, as shown in Figure 6, colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells are treated with isosakuranetin (Isosakuranetin), p53 known as a tumor suppressor expression was shown 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. The expression of both Cyclin D1 and CDK4 involved in the G1 phase was decreased, and the expression of pRB was also decreased. Due to this, it was found that the expression of Cyclin E and Cyclin A and CDK2 involved in the 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. (Isosakuranetin) has an anticancer effect and proves that it has an activity to inhibit cancer growth, recurrence and metastasis.

Example 4. Induction of apoptosis of cancer cells by treatment with Isosakuranetin

To evaluate the apoptosis changes of cancer cells following isosakuranetin treatment, colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were subjected to isosakuranetin (Isosakuranetin). ) was treated and the expression level of apoptosis-related proteins was measured.

As shown in Figure 7, colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells are treated with isosakuranetin (Isosakuranetin), the cleavage of PARP, a marker of apoptosis It was observed, and it was found that progenitor cell death factors such as Bim increased, and the expression of Bcl-2, an antiapoptotic 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. (Isosakuranetin) has an anticancer effect and proves that it has an activity to kill cancer cells.

Example 5. Induction of autophagy of cancer cells by treatment with Isosakuranetin

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

As shown in FIG. 8 , autophagy was induced on a fluorescence microscope by treatment with isosakuranetin in colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells. appear. In addition, as shown in Figure 9, colorectal cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells are treated with isosakuranetin (Isosakuranetin), the autophagy-related marker LC3B expression was shown to be induced.

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

Example 6. AKT-mTOR pathway changes in cancer cells according to isosakuranetin treatment

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

As shown in FIG. 10, as the colon cancer (HCT116) cells, lung cancer (A549) cells, and breast cancer (MDA-MB-231) cells were treated with isosakuranetin, p-Erk, p-AKT (Ser473) ), p-mTOR (Ser2448), p-P70S6K (Thr421/Ser424), and p-S6RP expression were decreased.

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

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, 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. According to claim 1, wherein the cancer is colon cancer, lung cancer, breast cancer, stomach cancer, liver cancer, pancreatic cancer, uterine cancer, esophageal cancer, bladder cancer, head and neck cancer, leukemia, lymphoma, blood cancer, rectal cancer, prostate cancer, ovarian cancer, kidney cancer, brain cancer , or a pharmaceutical composition characterized in that it is skin cancer. According to claim 1, wherein the isosakuranetin (Isosakuranetin) is a pharmaceutical composition, characterized in that it has an activity of inhibiting the growth of cancer by inhibiting the proliferation of cancer cells or by stopping the cell cycle of cancer cells in the G1 phase. The pharmaceutical composition according to claim 1, wherein the isosakuranetin has an activity to kill cancer cells by inducing apoptosis or autophagy of cancer cells. A health functional food composition for preventing or improving cancer comprising isosakuranetin or an acceptable salt thereof as an active ingredient as a food ingredient. According to claim 5, wherein the cancer is colon cancer, lung cancer, breast cancer, stomach cancer, liver cancer, pancreatic cancer, uterine cancer, esophageal cancer, bladder cancer, head and neck cancer, leukemia, lymphoma, blood cancer, rectal cancer, prostate cancer, ovarian cancer, kidney cancer, brain cancer , or a health functional food composition characterized in that it is skin cancer. A pharmaceutical composition for inhibiting cancer recurrence or metastasis, comprising isosakuranetin or a pharmaceutically acceptable salt thereof as an active ingredient. According to claim 7, wherein the isosakuranetin (Isosakuranetin) by inhibiting the proliferation of cancer cells or by stopping the cell cycle of cancer cells in the G1 phase, a pharmaceutical characterized in that it has an activity of inhibiting cancer recurrence and metastasis enemy composition.

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