KR20220003917A - Pharmaceutical composition for preventing or treating cancers comprising the gold nanoparticle of Crisiumjaponicum - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer, containing gold nanoparticles containing a Cirsium japonicum extract. More specifically, the present invention relates to a pharmaceutical composition comprising gold nanoparticles containing a Cirsium japonicum extract having an anti-cancer effect. The pharmaceutical composition comprising the gold nanoparticles containing the Cirsium japonicum extract according to the present invention has an effect of killing cancer cells and inhibiting survival, and exhibits superior anticancer activity compared to conventional anticancer drugs, thereby being able to be used as an anticancer therapeutic agent.

Description

TECHNICAL FIELD [0001] Pharmaceutical composition for preventing or treating cancers comprising the gold nanoparticles of Crisiumjaponicum, comprising gold nanoparticles containing milk thistle extract

The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer, comprising gold nanoparticles containing a milk thistle extract, and more particularly, to a pharmaceutical composition comprising gold nanoparticles containing a milk thistle extract having an anticancer effect on gastric cancer. to the composition.

Gastric cancer is the fourth most common cancer in the world and a common cause of cancer-related deaths. Although gastric cancer can be cured at an early stage, more than half of patients are diagnosed with gastric cancer at an advanced stage. Although various methods have been tried to treat gastric cancer, gastric cancer cells often resist such treatment. Statistically, the total cost of treating gastric cancer in the United States alone is expected to reach $2.31 billion in 2020, which imposes a significant cost burden, requiring the development of new drugs for gastric cancer treatment.

Among various therapies for treating cancer, inducing apoptosis is important, and apoptosis has several forms, such as apoptosis or feroptosis. Conventional apoptosis induction is mainly limited to apoptosis, but as apoptosis-related apoptosis, there is peroptosis, which may also play an important role in inducing apoptosis in cancer cells.

On the other hand, thistle ( Crisiumjaponicum. maackii ) plant is a perennial herb, and it is one of the herbs frequently used in China as well as Japan and Korea. Milk thistle has been reported to have medicinal, antidiabetic, antitumor, antioxidant and anti-inflammatory effects. For example, milk thistle extract has been used to treat Weil's disease and chronic hepatitis.

The main active ingredients of thistle plants include flavonoids, triterpenes, polyolefins, and some acetylenes, and although these active ingredients exhibit the above effects, these active ingredients are often found in diglycosylated form and have low hydrophobicity. Therefore, it is easy to pass through the biofilm, but it is in a form that is not soluble in water, so it is difficult to formulate, and there are problems that development and bioavailability as a drug are low (J. Agric. Food Chem. 2009, 57, 15, 6642-6648). To solve these problems, it is urgent to discover a technology for delivering extracts of plants of the genus Thistle to the body.

Accordingly, as a result of research efforts to develop a composition capable of reducing intracellular toxicity and enhancing anticancer effect by supporting a natural extract, the present inventors completed the development of gold nanoparticles containing a milk thistle extract capable of exhibiting anticancer activity.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, comprising gold nanoparticles including milk thistle extract as an active ingredient.

Also, the present invention

a) obtaining a thistle extract from milk thistle;

b) reacting the thistle extract with gold to obtain a mixture; and

c) centrifuging the mixture; An object of the present invention is to provide a method for preparing gold nanoparticles comprising a milk thistle extract, characterized in that it comprises a.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising gold nanoparticles containing milk thistle extract as an active ingredient.

In one embodiment of the present invention, the cancer may be any one selected from the group consisting of uterine cancer, breast cancer, stomach cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, blood cancer and liver cancer.

In another embodiment of the present invention, the composition may be characterized by increasing the expression of BAX, chtochrome C, Caspase 3 and Caspase 9.

In another embodiment of the present invention, the composition may be characterized in that it reduces the expression of GPX-4 and Bcl-2.

Also, the present invention

a) obtaining a thistle extract from milk thistle;

b) reacting the thistle extract with gold to obtain a mixture; and

c) centrifuging the mixture; It provides a method for producing gold nanoparticles comprising a milk thistle extract, characterized in that it comprises a.

In one embodiment of the present invention, in step b), the concentration of the milk thistle extract may be 0.1 mg/mL to 1 mg/mL, and the concentration of gold may be 0.5 mM to 3.5 mM.

In another embodiment of the present invention, in step b), the reaction may be carried out for 30 minutes to 50 minutes at 70 °C to 90 °C and pH 4 to pH 10 conditions.

The pharmaceutical composition comprising gold nanoparticles containing the milk thistle extract according to the present invention has cancer cell death and survival inhibitory effects, and exhibits superior anticancer activity compared to conventional anticancer drugs, and thus can be used as an anticancer therapeutic agent.

However, the effect of the present invention is not limited to the above effect, and it should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

1 shows detailed conditions for preparing gold nanoparticles containing milk thistle extract.
2 is a transmission electron microscope analysis result of gold nanoparticles containing a milk thistle extract.
3 is an energy-dispersive X-ray spectroscopy analysis result of gold nanoparticles containing a milk thistle extract.
4 is a result of particle size distribution analysis of gold nanoparticles containing milk thistle extract.
5 is an X-ray diffraction analysis result of gold nanoparticles containing milk thistle extract.
6 is an electron diffraction analysis result of gold nanoparticles containing milk thistle extract.
7 is a Fourier transform infrared analysis result of gold nanoparticles containing milk thistle extract.
8 is a result showing the cell viability when treated with gold nanoparticles containing milk thistle extract for Raw264.7 cell line, NHDF cell line, and AGS cell line.
9 is a microscopic view of an AGS cell line treated with gold nanoparticles containing a milk thistle extract.
10 is a result showing cell morphology, colony formation, and wound healing assays that appear after each treatment of gold nanoparticles and thistle extract containing a milk thistle extract in AGS cell lines.
11 is a result of lysosome formation in AGS cell lines after each treatment with gold nanoparticles containing a milk thistle extract and a milk thistle extract.
12 is a result of the formation of mitophagosomes after the AGS cell line was treated with gold nanoparticles containing a milk thistle extract and a milk thistle extract, respectively.
13 is a result confirming the apoptosis-inducing effect of gold nanoparticles containing control, cisplatin, gold nanoparticles, milk thistle extract and thistle extract on AGS cells through propidium iodide staining and Hoechst33258 staining.
14 is a comparison result of mRNA expression levels of apoptosis-related genes BAX, BCL-2, Caspase3, Caspase9, and Cytocrome C, which appear when milk thistle extract and gold nanoparticles containing milk thistle extract are treated in AGS cell lines.
15 is a graph showing the cell viability of AGS cell lines treated with milk thistle extract, gold nanoparticles containing milk thistle extract, Erastin, and Ferrostain-1, respectively, or a mixture of each concentration.
16 is a result confirming the effect of inducing feroptosis when AGS cell lines are treated with thistle extract, gold nanoparticles containing thistle extract, Erastin, and Ferrostain-1, respectively or by mixing them.
17 is a comparison result of the protein expression level of GPX-4 when AGS cell lines were treated with thistle extract, gold nanoparticles containing thistle extract, Erastin, and Ferrostain-1, respectively.

The present inventors conducted research to develop a composition capable of reducing intracellular toxicity and enhancing anticancer effect. As a result, the development of gold nanoparticles containing thistle extract with superior anticancer activity compared to conventional anticancer drugs and thistle extract was completed.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating cancer, comprising gold nanoparticles containing milk thistle extract as an active ingredient.

In the present invention, the " milk thistle extract" refers to a substance obtained by concentrating a plant belonging to the Thistle family belonging to the genus Crisium japonicum , and is known to have anti-inflammatory, antioxidant and anticancer effects.

In the present invention, the extract can be extracted according to a conventional method known in the art for extracting an extract from a natural product, that is, using a conventional solvent under conditions of a conventional temperature and pressure. For example, in the present invention, the extract may use one or more solvents selected from water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof. In addition, the method of extracting the extract may be extracted through various methods such as hot water extraction, cold extraction extraction, reflux extraction, ultrasonic extraction, but is not limited thereto.

The prepared extract may then be filtered or concentrated or dried to remove the solvent, and both filtration, concentration and drying may be performed. For example, the filtration may be performed using filter paper or a reduced pressure filter, the concentration may be performed by a vacuum concentrator, and the drying may be performed by a freeze-drying method, but is not limited thereto.

As used herein, the term “cancer” refers to a disease in which cells proliferate unrestrictedly in living tissues to cause malignant tumors, and is divided into carcinomas generated from epithelial cells and sarcomas generated from non-epithelial cells. Cancer is divided into gastric cancer, liver cancer, lung cancer, pancreatic cancer, etc. according to the site of occurrence, and there are various types, and each type of cancer has different treatment methods and effects.

In the present invention, the target cancer for which the milk thistle extract has an anticancer effect may be any one selected from the group consisting of uterine cancer, breast cancer, stomach cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, blood cancer and liver cancer, preferably It could be stomach cancer.

In the present invention, "a pharmaceutical composition for preventing or treating cancer" refers to a substance that inhibits the proliferation of cancer cells or kills cancer cells, and the inhibition or death of cancer cells causes cytotoxicity to cancer cells due to the composition, decomposing cancer cells. It means to increase the expression of factors related to the formation of mitophagosomes and lysosomes or cell death.

Accordingly, in the present invention, "cancer prevention or treatment" preferably increases the expression of BAX, chtochrome C, Caspase 3 and Caspase 9, which are factors related to apoptosis, and the expression of Bcl-2, which is known to inhibit apoptosis. and reducing the expression of GPX-4, which is known to inhibit peroptosis.

The present inventors confirmed that the composition of the present invention has a cytotoxic effect and apoptosis effect on cancer cells through specific examples.

Specifically, in one embodiment of the present invention, gold nanoparticles and thistle extract containing milk thistle were treated in the Raw264.7 cell line, NHDF cell line, and AGS cell line, which is a human gastric cancer cell line, which are mouse macrophages, respectively. Cell viability of Raw264.7 cell line and NHDF cell line was not different when treated with gold nanoparticles containing milk thistle compared to treatment with milk thistle extract, but the viability of AGS cells was significant when treated with gold nanoparticles containing thistle extract compared to this extract It was confirmed to decrease significantly. Through this, it was confirmed that the case where the milk thistle extract was supported on the gold nanoparticles had a significantly superior anticancer effect compared to the case where it was not. In addition, it was confirmed that treatment with gold nanoparticles containing milk thistle extract showed significantly superior cell proliferation inhibitory effect on AGS cells compared to the case where AGS cells were treated with thistle extract and cisplatin, which is known as a conventional anticancer agent, respectively ( See Example 3-1).

In another embodiment of the present invention, the treatment of gold nanoparticles containing milk thistle extract on AGS cells compared to the treatment of gold nanoparticles, cisplatin and thistle extract respectively on AGS cells, mitophago associated with intracellular hydrolysis It was confirmed that the formation of some or lysosomes was significantly induced. In addition, treatment of gold nanoparticles containing milk thistle extract increased the expression of genes related to apoptosis induction, BAX, Caspase3, Caspase9 and cytochrome C, compared to treatment with cisplatin and milk thistle extract, respectively, and Bcl-, which is known to inhibit apoptosis It was confirmed that the expression of 2 was reduced. Accordingly, it was confirmed that the gold nanoparticles containing the thistle extract had a significantly superior apoptosis effect compared to the gold nanoparticles, cisplatin, and the milk thistle extract, respectively (see Example 3-2).

In another embodiment of the present invention, AGS cells contain thistle extract, gold nanoparticles containing milk thistle extract, gold nanoparticles containing thistle extract containing Erastin, a feroptosis inducer, and thistle extract containing Ferrostatin, an inhibitor of feroptosis. When gold nanoparticles were treated individually, the cell viability of AGS cells was significantly reduced in gold nanoparticles containing the same amount of milk thistle extract compared to milk thistle extract. Even in this case, it was confirmed that the cell viability was significantly lower than that of the milk thistle extract. In addition, it was confirmed that the treatment of gold nanoparticles containing milk thistle extract reduced the expression of GPX-4, which is known to inhibit peroptosis, compared to treatment with milk thistle extract and ferrostatin, respectively. Accordingly, it was confirmed that the gold nanoparticles containing the milk thistle extract had a significantly superior peroptosis-inducing effect compared to the milk thistle extract (see Example 3-3).

The pharmaceutical composition according to the present invention may further include a commonly used pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is commonly used in formulation, and includes saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol and liposomes, etc. It is not limited, and may further include other conventional additives, such as antioxidants and buffers, if necessary. In addition, diluents, dispersants, surfactants, binders, lubricants, etc. may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets. Regarding suitable pharmaceutically acceptable carriers and formulations, formulations can be preferably made according to each component using the method disclosed in Remington's literature. The pharmaceutical composition of the present invention is not particularly limited in the formulation, but may be formulated as injections, inhalants, external preparations for skin, and the like.

The composition of the present invention may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally, or locally applied) according to a desired method, but preferably may be administered orally, and the dosage may be determined by the patient. Although it varies depending on the condition and weight of the patient, the degree of disease, the drug form, the route and time of administration, it may be appropriately selected by those skilled in the art.

The composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat or diagnose a disease at a reasonable benefit/risk ratio applicable to medical treatment or diagnosis, and the effective dose level is determined by the patient's disease type, severity, drug activity, sensitivity to drugs, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs, and other factors well known in the medical field. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or may be administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art. Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on the patient's age, sex, condition, weight, absorption of the active ingredient into the body, inactivation rate and excretion rate, disease type, and drugs used in combination, in general 0.001 to 150 mg, preferably 0.01 to 100 mg per 1 kg of body weight, may be administered daily or every other day, or may be administered in divided doses 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, the severity of obesity, sex, weight, age, etc., the dosage is not intended to limit the scope of the present invention in any way.

In another aspect of the present invention, the present invention

a) obtaining a thistle extract from milk thistle;

b) reacting the thistle extract with gold to obtain a mixture; and

c) centrifuging the mixture; It provides a method for producing gold nanoparticles comprising a milk thistle extract, characterized in that it comprises a.

In the present invention, the concentration of the milk thistle extract used in the reaction may be 0.1 mg/mL to 1 mg/mL, preferably 0.25 mg/mL to 0.75 mg/mL, and more preferably 0.5 mg /mL.

In the present invention, the concentration of gold used in the reaction may be 0.5 mM to 3.5 mM, preferably 1 mM to 3 mM, and more preferably 2 mM.

In the present invention, the reaction temperature used for the reaction may be 70 °C to 90 °C, preferably 75 °C to 85 °C, more preferably 80 °C.

In the present invention, the reaction time used for the reaction may be 30 minutes to 50 minutes, preferably 35 minutes to 45 minutes, more preferably 40 minutes.

In the present invention, the pH used for the reaction may be 4 to 10, preferably 6 to 8, more preferably 7 may be.

In an embodiment of the present invention, 70% ethanol of 10 times the volume of thistle was added to dried thistle, extracted under reflux at 65° C. for 3 hours, and then vacuum filtration, concentration and drying oven were performed to prepare a thistle extract.

After that, the optimal conditions were checked by varying the concentration, reaction temperature, reaction time, gold concentration, and pH range of the thistle extract. As a result, the concentration of the milk thistle extract was 0.5 mg/mL, the reaction temperature was 80 °C, and the reaction time was 40 minutes. , it was confirmed that the yield of gold nanoparticles containing thistle extract was the highest when the gold concentration was 2 mM and pH 7 (see Example 1).

In another embodiment of the present invention, transmission election microscope (TEM), energy-dispersive X-ray spectroscpty (EDX), X-ray diffraction (XRD), electron The physical properties of the nanoparticles prepared in Example 1 were confirmed by using diffraction analysis (Selected area election eiffraction) and Fourier transform infrared (FTIR) to confirm the preparation of gold nanoparticles containing thistle extract. (See Example 2).

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these Examples and the like are intended to explain the present invention in more detail, it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereby.

[Example]

Example 1. Preparation of Gold Nanoparticles Containing Milk Thistle Extract

1-1. Thistle Extract Preparation

In order to prepare gold nanoparticles containing thistle extract according to the present invention, the following experiment was performed.

First, to prepare a thistle extract, dried thistle was purchased from a farmhouse in Imsil, Jeollabuk-do. 70% ethanol 10 times the volume of thistle was added to the dried thistle, and extraction was performed under reflux at 65° C. for 3 hours. The extracted milk thistle extract was vacuum filtered at 25° C., and the filtrate was concentrated at 45° C. using a vacuum rotary evaporator. The concentrate was dried in a drying oven at 40°C and then stored at -20°C for further experiments.

1-2. Identification of Optimal Conditions for Preparation of Gold Nanoparticles Containing Milk Thistle Extract

1-2-1. Milk Thistle Extract Concentration

As shown in FIG. 1 , gold containing thistle extract was obtained by varying the concentration of the thistle extract to 0.125 mg/mL, 0.25 mg/mL, 0.5 mg/mL, 1 mg/mL, 2 mg/mL and 4 mg/mL. As a result of confirming the yield of nanoparticles, it was found that the highest yield appeared when 0.5 mg/mL.

1-2-2. reaction temperature

As shown in FIG. 1 , the reaction temperature for preparing the nanoparticles containing the thistle extract was confirmed at 20°C to 100°C at 10°C intervals. When the reaction temperature was 80°C, the yield of gold nanoparticles containing the thistle extract was high. was found to be the highest.

1-2-3. reaction time

As shown in FIG. 1 , as a result of checking the reaction time for preparing the nanoparticles containing thistle extract from 10 minutes to 60 minutes at 10-minute intervals, when the reaction time was 40 minutes, the yield of gold nanoparticles containing the thistle extract was the highest. was confirmed to be high.

1-2-4. gold concentration

As shown in FIG. 1 , the concentration of gold used to prepare the gold nanoparticles containing the thistle extract was checked at 0.5 mM to 3.5 mM at 0.5 mM intervals. As a result, when the concentration of gold used was 2 mM, the thistle extract contained It was confirmed that the yield of gold nanoparticles was the highest.

1-2-5. reaction pH

As shown in FIG. 1 , when the reaction pH for preparing the gold nanoparticles containing the thistle extract was checked at 2 to 7 at 1 interval, when the reaction pH was 7, the yield of gold nanoparticles containing the thistle extract was the highest. Confirmed.

The mixture obtained under the above conditions was centrifuged at 1,2000 rpm for 20 minutes to finally obtain gold nanoparticles containing thistle extract. The gold nanoparticles containing the obtained thistle extract were washed with distilled water and then dried in the air.

Hereinafter, an experiment was conducted with gold nanoparticles containing the thistle extract obtained under the optimum conditions of 0.5 mg/mL of the thistle extract, a reaction temperature of 80° C., a reaction time of 40 minutes, a gold concentration of 2 mM, and a pH of 7.

Example 2. Characterization of Gold Nanoparticles Containing Milk Thistle Extract

In order to confirm whether the preparation of the gold nanoparticles containing the thistle extract according to the present invention was well performed, as shown in FIGS. 2 to 7 , transmission election microscope (TEM), energy dispersive X-ray spectroscopy (Energy -dispersive X-ray spectroscpty (EDX), X-ray diffraction (XRD), electron diffraction analysis (Selected area election eiffraction), and Fourier transform infrared (FTIR) in Example 1 The physical properties of the nanoparticles prepared in .

As shown in FIG. 2 , the surface of the gold nanoparticles containing the thistle extract and dispersion of the gold particles could be confirmed through electron microscopic analysis. Through this, it was possible to confirm the preparation of gold nanoparticles containing the thistle extract in which gold particles were evenly distributed.

Also, as shown in FIG. 3 , the presence of gold particles could be confirmed by energy dispersive X-ray spectroscopy, and as shown in FIG. 4 , the gold nanoparticles containing thistle extract produced by particle size distribution were The number, density and volume distribution were confirmed. Through this, it was confirmed that the material mixed with the thistle extract was gold nanoparticles, and gold nanoparticles containing the thistle extract were prepared.

In addition, as shown in FIGS. 5 and 6, the structure of the prepared nanoparticles was confirmed through X-ray diffraction analysis and electron diffraction, and in FIG. 7, between gold thistle extract and gold nanoparticles containing thistle extract through Fourier transform infrared. It was confirmed that there was a difference in molecular structure.

By performing the above experiment, it was confirmed that the gold nanoparticles containing the milk thistle extract in Example 1 were well prepared.

Example 3. Confirmation of anticancer activity of gold nanoparticles containing milk thistle extract

3-1. Confirmation of inhibition of cancer cell proliferation

In order to confirm the cytotoxic effect of gold nanoparticles containing the milk thistle extract prepared in Example 1 on gastric cancer cells, the nanoparticles were added to the mouse macrophage Raw264.7 cell line, the NHDF cell line, and the human gastric cancer cell line AGS cell line. Treatment was performed to measure the viability of the cells.

More specifically, the Raw264.7 cell line, the NHDF cell line and the AGS cell line were cultured in DMEM and RPMI medium supplemented with 10% FBS and 1% penicillin-streptomycin (5% CO ₂ , 37° C.). Each of the cultured Raw264.7 cell line, NHDF cell line, and AGS cell line was treated with thistle extract and gold nanoparticles containing thistle extract at concentrations of 100, 150 and 200 μg/ml, respectively, and cultured for 24 hours, followed by MTT assay. Cytotoxicity and cell growth rates were measured. As a result, as shown in FIG. 8 , in the Raw264.7 cell line and the NHDF cell line, there was no significant difference in cell viability when the gold nanoparticles containing the thistle extract and the thistle extract were treated, respectively, but in the case of the AGS cell line, the gold nanoparticles containing the thistle extract Cell viability was significantly reduced compared to milk thistle extract when treated with particles.

Also, as shown in FIG. 9 , it was confirmed that AGS cell proliferation was inhibited in gold nanoparticles containing milk thistle extract compared to the control group when observed under a microscope.

Also, as shown in FIG. 10 , when gold nanoparticles containing cisplatin, thistle extract, and thistle extract, known as anticancer drugs, were administered to AGS cells, respectively, and wound healing assay was performed, the gold nanoparticles containing thistle extract were compared to other controls. It appeared that cell proliferation appeared in a slow form, colony formation was small, and cell activity and migration rate were suppressed.

From the above results, it was confirmed that gold nanoparticles containing milk thistle extract had a significantly superior cancer cell proliferation inhibitory effect on gastric cancer cells than cisplatin, gold nanoparticles, and milk thistle extract. In particular, it was found that when the milk thistle extract was supported on gold nanoparticles, it had a significantly superior anticancer effect compared to the case where it was not.

3-2. Confirmation of apoptosis effect

In order to confirm the gastric cancer cell killing effect of the gold nanoparticles containing the milk thistle extract prepared in Example 1, AGS cells were treated with the gold nanoparticles containing the milk thistle extract, followed by intracellular hydrolysis-related mitophagosome or lysosome It was confirmed whether the formation of

As a result, as shown in FIGS. 11 and 12 , when gold nanoparticles containing thistle extract, gold nanoparticles, cisplatin, and milk thistle extract were treated in AGS cells, respectively, mitophagosome and mitophagosome and gold nanoparticles containing thistle extract It was confirmed by fluorescence microscopy that the effect of inducing lysosome formation was markedly excellent.

In addition, as shown in FIG. 13 , apoptosis was measured after treatment of gold nanoparticles containing thistle extract, cisplatin, gold nanoparticles, and thistle extract to AGS cells through propidium iodide staining and Hoechst33258 staining. It was confirmed that the particles induce a significantly superior apoptotic effect compared to gold nanoparticles, milk thistle extract, and cisplatin.

In addition, as shown in FIG. 14 , as a result of treating AGS cells with gold nanoparticles containing thistle extract, cisplatin, and milk thistle extract, respectively, a gene related to apoptosis in the gold nanoparticles containing thistle extract compared to cisplatin and thistle extract was found. The expression of BAX, Caspase3, Caspase9 and cytochrome C was increased, and the expression of Bcl-2, which is known to inhibit apoptosis, was decreased.

Through this, it can be seen that gold nanoparticles containing milk thistle extract have a significantly superior apoptotic effect on gastric cancer cells compared to conventional anticancer drugs and milk thistle extract.

3-3. Confirmation of the effect of iron-dependent apoptosis (peroptosis)

In order to confirm the effect of iron-dependent apoptosis on gastric cancer cells by gold nanoparticles containing milk thistle extract, gold nanoparticles containing milk thistle extract, Erastin, a feroptosis inducer, and Ferrostain-1, an inhibitor, were added to AGS cells, respectively. After treatment with thistle extract-containing gold nanoparticles, milk thistle extract, Ferrostain-1 and Erastin, it was checked whether feroptosis was induced.

As shown in FIGS. 15 and 16 , the cell viability of AGS cells was significantly reduced in the gold nanoparticles containing the milk thistle extract in the same amount as compared to the milk thistle extract, and Ferrostain-1, a peroptosis inhibitor, was added to the gold nanoparticles containing the thistle extract. Even when added, the cell viability was significantly lower than that of the milk thistle extract. As a result of treating the gold nanoparticles containing the milk thistle extract with Erastin, a peroptosis inducer, it was confirmed that the cell viability was significantly reduced.

Also, as shown in FIG. 17 , as a result of treating AGS cells with gold nanoparticles containing thistle extract, thistle extract, Ferrostain-1, and Erastin, respectively, gold nanoparticles containing thistle extract inhibited peroptosis compared to this extract. It was confirmed that the expression of GPX-4, which is known to be significantly reduced. In addition, it was confirmed that, as a result of treating AGS cells with gold nanoparticles containing milk thistle extract, GPX-4 expression similar to that of Erastin, a peroptosis derivative, appeared.

Through this, it can be seen that when the milk thistle extract is contained in gold nanoparticles and treated with the cells, it has a significantly superior peroptotic effect on gastric cancer cells compared to the milk thistle extract.

The stated description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (7)

A pharmaceutical composition for preventing or treating cancer, comprising gold nanoparticles containing milk thistle extract as an active ingredient.
The method of claim 1,
The cancer is uterine cancer, breast cancer, stomach cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, a pharmaceutical composition of any one selected from the group consisting of blood cancer and liver cancer.
The method of claim 1,
The composition is a composition, characterized in that it increases the expression of BAX, chtochrome C, Caspase 3 and Caspase 9.
The method of claim 1,
The composition is a composition, characterized in that it reduces the expression of Bcl-2 and GPX-4.
a) obtaining a thistle extract from milk thistle;
b) reacting the thistle extract with gold to obtain a mixture; and
c) centrifuging the mixture; A method for producing gold nanoparticles comprising a milk thistle extract, comprising:
6. The method of claim 5,
In step b), the concentration of the milk thistle extract is 0.1 mg/mL to 1 mg/mL, and the concentration of gold is 0.5 mM to 3.5 mM.
6. The method of claim 5,
In step b), the reaction is characterized in that made for 30 minutes to 50 minutes at 70 ℃ to 90 ℃ and pH 4 to pH 10 conditions, the manufacturing method.

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