KR20200098311A - Pharmaceutical composition for preventing or treating glioblastoma comprising Phellodendron amurense Ruprecht or Scutellaria baicalensis - Google Patents

Abstract

The present invention relates to a composition for preventing or treating glioblastoma including an extract of Scutellaria baicalensis, Phellodendron amurense Ruprecht or Rubus coreanus, or a fraction thereof, as an active ingredient. More particularly, the present invention relates to a pharmaceutical composition, a food composition and a feed composition for preventing or treating glioblastoma including an extract of Scutellaria baicalensis, Phellodendron amurense Ruprecht or Rubus coreanus, or a fraction thereof, as an active ingredient; and a method for preventing or treating glioblastoma including a step of administering the pharmaceutical composition. According to the present invention, Scutellaria baicalensis, Phellodendron amurense Ruprecht or Rubus coreanus shows an excellent effect of treating glioblastoma, and can be used for a pharmaceutical composition or food to develop an agent for treating or alleviating glioblastoma.

Description

Composition for preventing or treating glioblastoma comprising herbal extracts {Pharmaceutical composition for preventing or treating glioblastoma comprising Phellodendron amurense Ruprecht or Scutellaria baicalensis}

The present invention relates to a composition for preventing or treating glioblastoma comprising a golden, hwangbaek or bokbunja extract or a fraction thereof as an active ingredient, and more specifically, the present invention comprises a golden, hwangbaek or bokbunja extract or a fraction thereof as an active ingredient It relates to a method for preventing or treating glioblastoma comprising the step of administering a pharmaceutical composition, a food composition, a feed composition, and the pharmaceutical composition for preventing or treating glioblastoma.

Glioblastoma (GBM, glioblastoma multiforme) refers to the most malignant glioma of WHO classification 4 among primary brain cancers and is known to have a very poor prognosis. The standard treatment for glioblastoma patients is surgical resection, radiotherapy, and chemotherapy using temozolomide, but the average survival rate is only 14 months, so more effective treatment is required.

Axl receptor tyrosine kinase belongs to the group of TAM (Tyro3-Axl-Mertk) receptors, and has recently attracted attention as a target protein for cancer treatment. The protein structure consists of two immunoglobulin-like (Ig) domains and two fibronectin III domains exposed outside the cell, a single pass transmembrane domain and a tyrosine phosphorylation domain inside the cell. Growth arrest-specific gene 6 (GAS6) and Protein S (PROS1) are known to bind to the Axl receptor and induce activation.

Clinically, it was reported that Axl expression was higher in primary and metastatic cancer compared to normal tissues, and the higher the Axl expression was, the poorer the prognosis was in lung cancer, pancreatic cancer, kidney cancer, colon cancer, liver cancer, esophageal cancer, and glioblastoma. In addition, Axl is known as a protein that plays an important role in inducing intrinsic and acquired resistance to chemotherapy, immunotherapy, and molecular target anticancer agents. Since the first report of a study showing that Axl mRNA is more than twice as high in cisplatin-resistant ovarian cancer, it has been found that there is a correlation between Axl expression and anticancer drug resistance in breast cancer, colon cancer, and lung cancer. Recently, it has been found that overexpression of Axl is a mechanism of resistance to erlotinib, an anticancer drug targeting EGFR, in non-small cell lung cancer with a mutation in the Epidermal growth factor (EGF) receptor.

As the results of recent studies prove that Axl plays a very important role in cancer cell growth and resistance to anticancer drugs in almost all carcinomas, anticancer agents targeting Axl have been developed, and clinical trials for this are being actively conducted. The fact that Axl inhibitors induce cancer cell death and that Axl inhibition is effective as a method to overcome anticancer drug resistance to PARP inhibitors and PI3K inhibitors was found to be effective.

As such, the importance of Axl in cancer research has been greatly emphasized through the research results for the past 10 years. In particular, research results that Axl inhibitors inhibit the growth, invasion, and metastasis of glioblastoma have been steadily published. There is no development of natural anticancer drugs.

Under this background, the present inventors have made extensive research efforts to discover an herbal medicine that inhibits the growth of glioblastoma by inhibiting the expression of Axl targeting Axl, and as a result, it has been confirmed that gold, Hwangbaek and bokbunja have an effect on glioblastoma inhibition, The present invention has been completed.

One object of the present invention is to provide a pharmaceutical composition for preventing or treating glioblastoma comprising any one or more extracts or fractions thereof selected from the group consisting of golden, yellow and red bokbun as an active ingredient.

Another object of the present invention is to provide a food composition for preventing or improving glioblastoma, comprising any one or more extracts selected from the group consisting of gold, yellow white and bokbun, or a fraction thereof as an active ingredient.

Another object of the present invention is to provide a feed composition for preventing or improving glioblastoma comprising any one or more extracts selected from the group consisting of gold, yellow white and bokbunja or a fraction thereof as an active ingredient.

Another object of the present invention is to provide a method for preventing or treating glioblastoma comprising administering the pharmaceutical composition.

One embodiment of the present invention for achieving the above object is to provide a pharmaceutical composition for preventing or treating glioblastoma comprising any one or more extracts selected from the group consisting of golden, yellow, and bokbun as an active ingredient or a fraction thereof.

The term "golden (Scutellaria baicalensis)" of the present invention refers to a medicinal material made from the roots of gold (Scutellaria baicalensis GEORGE), a perennial herbaceous plant belonging to the Lamiaceae family. The gold generally grows in the grassy field of the mountain and grows as a vegetation with several generations, hairs, and branches are split. The root is conical and the flesh is yellow. In herbal medicine, the root is used as antipyretic, diuretic, branch, ear infection, and anti-inflammatory. On the other hand, baicalin, a kind of flavonoid contained in the gold, is known to have antioxidant, anticancer, and anti-inflammatory effects.

The term "Hwangbaek (Phellodendron amurense Ruprecht)" of the present invention refers to the bark of the Hwangbyeok tree used as a medicinal material, and is also called Hwanggyeongpi. It is a deciduous broad-leaved arboreous tree that grows up to 10m in height. Cork is developed on the bark and the inner bark is yellow. In Korea, it is evenly distributed in all areas except Jeju and Jeonnam, and grows in a mixed forest or valley in the mountains. The Hwangbaek is used as a medicinal material by peeling the bark from a tree trunk before and after the lower extremities to remove or chop it and dry it in the sun. The Hwangbaek is known to lower blood sugar, inhibit the growth of pneumococcal, tuberculosis, staphylococcus, and the like, and prevent the growth of tumor cells, and sterilize.

The term "Rubus coreanus" in the present invention belongs to the Rosaceae family, and is known to have origin in China, and Jeju Island and southern regions are known as major production areas in Korea. Bokbunja harvests dark red fruits in early summer and uses them for edible use.Since ancient times, it has been known as a valuable medicinal material as a tonic, gangjeong, and a summary that protects the liver, and is known to be used in physical weakness, yangwi, oil and urine.

The term “extract” of the present invention refers to a substance obtained by extracting a substance, and specifically, an extract obtained by the extraction treatment of the present invention, a dilution or concentrate of the extract, or a dried product obtained by drying the extract , It includes the extract of all formulations that can be formed by using the extract itself and the extract, such as the preparation or purified product of the extract or a mixture thereof.

The usual extract may be a single extract of golden, yellow white or bokbunja, and may be a complex extract of two or more selected from the group consisting of golden, yellow and red, but is not limited thereto. More specifically, golden and yellow white extract; Hwangbaek and bokbunja extract; Golden and bokbunja extract; Or golden, hwangbaek and bokbunja extract; Can be

The method of extracting the extract is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extract extraction method include a solvent extraction method, an ultrasonic extraction method, a filtration method, a reflux extraction method, and the like, and these may be performed alone or in combination of two or more methods.

In the present invention, the type of the extraction solvent used for the extract is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and these may be used alone or in combination of one or more. Specifically, the extraction solvent may be hot water.

As used herein, the term "fraction" means a result obtained by performing fractionation in order to separate a specific component or a specific group of components from a mixture containing various components.

The fractionation method for obtaining the fraction in the present invention is not particularly limited as long as it can exhibit an effect of inhibiting glioblastoma, but may be performed according to a method commonly used in the art. For example, a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing through an ultrafiltration membrane having a certain molecular weight cut-off value, and various chromatography (separation according to size, charge, hydrophobicity or affinity) It may be a chromatographic fractionation method and a combination thereof, etc. to perform one designed for).

In the present invention, the kind of the fractionation solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvent include polar solvents such as water, distilled water, and alcohol; Non-polar solvents, such as hexane (Hexan), ethyl acetate (Ethyl acetate), chloroform (Chloroform), and dichloromethane (Dichloromethane), etc. are mentioned. These may be used alone or in combination of two or more. In the case of using an alcohol in the fractionation solvent, preferably a C1 to C4 alcohol may be used.

The term "prevention" of the present invention means any action of inhibiting or delaying the growth of glioblastoma by administration of the pharmaceutical composition according to the present invention.

The term "treatment" of the present invention refers to any action in which symptoms of a suspicious and onset individual of glioblastoma are improved or beneficially altered by administration of the pharmaceutical composition.

The content of the golden, yellow white or bokbunja extract contained in the pharmaceutical composition of the present invention is not particularly limited thereto, but 0.0001 to 80% by weight, 0.0001 to 50% by weight, more specifically 0.01 to 20% by weight based on the total weight of the final composition It can be included in% content.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent, and the carrier may include a non-naturally occurring carrier.

More specifically, the carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate. , Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, polycaprolactone, polylactic acid (Poly Lactic Acid), poly-L-lactic acid, mineral oil, etc. are mentioned.

Each of the pharmaceutical compositions may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions according to a conventional method, The form of the carrier may include various amorphous carriers, microspheres, nanofibers, and the like.

In the case of formulation, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient, such as starch, calcium carbonate, in the extract and its fractions, It can be prepared by mixing sucrose or lactose, gelatin, and the like. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. have.

Preparations for parenteral administration may include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, suppositories, and the like. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used.

Another embodiment of the present invention provides a method for preventing or treating glioblastoma comprising administering the pharmaceutical composition to an individual.

At this time, the description of the “pharmaceutical composition”, “prevention” and “treatment” is as described above.

Since the pharmaceutical composition of the present invention exhibits a prophylactic or therapeutic effect of glioblastoma, the method of the present invention including the step of administering it to an individual can be usefully utilized for the prevention or treatment of glioblastoma.

The term "administering" of the present invention means introducing the composition to a subject in an appropriate manner.

The term "individual" of the present invention refers to all animals such as mice, mice, livestock, etc., including humans that have or may develop glioblastoma, and may be mammals including humans, but is not limited thereto. .

Another embodiment of the present invention provides a food composition for preventing or improving glioblastoma, comprising as an active ingredient any one or more extracts selected from the group consisting of gold, yellow white and bokbun.

In this case, the definitions of the terms “golden”, “hwangbaek”, “bokbunja”, “extract”, “fraction” and “prevention” are as described above.

The term "improvement" of the present invention means any action that at least reduces the severity of a parameter, such as a symptom, related to the condition being treated with the administration of the composition.

Since the golden, hwangbaek or bokbunja extract according to the present invention exhibits an excellent glioblastoma inhibitory effect, it can be included in a food composition for the purpose of preventing or improving glioblastoma, and the food composition can be consumed on a daily basis. High effects can be expected for the prevention or improvement of

In the present invention, the term "food" refers to meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages , Vitamin complexes, health functional foods, etc., including all foods in the usual sense, and as long as it may contain the golden or yellow white of the present invention, it is not limited thereto.

The term "health functional food" of the present invention refers to a food manufactured and processed using raw materials or ingredients having useful functions for the human body according to the Health Functional Food Act No.6727, and'functionality' refers to the structure of the human body. And it means to obtain a useful effect for health use, such as controlling nutrients for function or physiological action. On the other hand, health food refers to food that has an active health maintenance or promotion effect compared to general food, and health supplement food refers to food for the purpose of health supplement. In some cases, the terms of health functional food, health food, and health supplement food Can be used interchangeably.

Golden, yellow white or bokbunja of the present invention may be added as it is or may be used together with other foods or food ingredients, and may be appropriately used according to a conventional method.

The food product of the present invention can be prepared by a method commonly used in the art, and during the production, raw materials and ingredients commonly added in the art may be added to prepare it. Specifically, the food composition may further include a physiologically acceptable carrier, and the type of the carrier is not particularly limited, and any carrier commonly used in the art may be used. In addition, the food composition may contain food additives such as preservatives, disinfectants, antioxidants, coloring agents, coloring agents, bleaching agents, seasonings, sweeteners, flavoring agents, expanding agents, reinforcing agents, emulsifying agents, thickening agents, coating agents, gum base agents, foam inhibitors, solvents, and improving agents. Can include. The additive may be selected according to the type of food and used in an appropriate amount.

In addition, the formulation of the food may be prepared without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be prepared in various forms of formulation, and unlike general drugs, it has the advantage of not having side effects that may occur during long-term use of drugs using food as a raw material, and is excellent in portability. Food can be taken as an adjuvant to enhance the effect of preventing or improving glioblastoma.

Golden, Hwangbaek, or bokbunja of the present invention may be included in a variety of weight percent in the food composition if it can exhibit the effect of preventing or improving glioblastoma. Specifically, it may be included in 0.00001 to 100% by weight or 0.01 to 80% by weight based on the total weight of the food composition, but is not limited thereto. When ingested for a long time for health and hygiene purposes, the content below the above range may be included, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

Another embodiment of the present invention provides a feed composition for preventing or improving glioblastoma, comprising as an active ingredient any one or more extracts selected from the group consisting of golden, hwangbaek, and bokbun.

At this time, the definitions of the terms “golden”, “hwangbaek”, “bokbunja”, “extract”, “fraction”, “prevention” and “improvement” are as described above.

Golden, Hwangbaek, or Bokbunja according to the present invention exhibits excellent glioblastoma treatment effect, so it can be included in the feed composition for the purpose of preventing or improving glioblastoma, and the feed composition can be consumed on a daily basis by animals. High effects can be expected in the prevention or improvement of blastoma.

The term "feed" of the present invention means any natural or artificial diet, one meal meal, etc., or a component of the one meal meal, for animals to eat, ingest, and digest.

The kind of feed is not particularly limited, and feed commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, meals or grain by-products; Animal feeds such as proteins, inorganic logistics, oils and fats, minerals, oils and fats, single-cell proteins, zooplanktons or foods. These may be used alone or in combination of two or more.

Since the golden, hwangbaek or bokbunja of the present invention exhibits excellent glioblastoma treatment effect, it can be included as an active ingredient in a pharmaceutical composition or food and used in the development of a therapeutic agent or ameliorating agent for glioblastoma.

1 is a graph comparing the effect of Axl on glioblastoma growth by loss of Axl gene through shRNA in glioblastoma U87MG and U373MG cells through MTT analysis.
FIG. 2 is a graph analyzed by Western blotting to determine which mechanism was inhibited in glioblastoma U87MG and U373MG in which the Axl gene was lost, thereby inhibiting the growth of glioblastoma.
Figure 3 is a graph showing the result of comparing the expression amount of Axl and GAPDH through Western blotting after treating 26 kinds of herbal medicine water extracts to U87MG, a glioblastoma, for 24 hours in order to find an herbal medicine that can inhibit Axl to be.
Figure 4 is a graph showing the results of fertilizing the growth of the cells by treating the Hwangbaek extract showing the effect of inhibiting Axl in U87MG, a glioblastoma, for 48 hours.
5 is a graph showing the results of fertilizing the growth of the cells by treating the golden extract, which showed the effect of inhibiting Axl, in U87MG, a glioblastoma, for 48 hours.
6 is a graph analyzed through Western blotting to determine which mechanisms of gold and yellow white, which showed the effect of inhibiting the growth of glioblastoma, inhibited the growth of glioblastoma.

Hereinafter, the present invention will be described in more detail by the following examples. However, the following examples are for illustrative purposes only, and the scope of the present invention is not limited thereto.

Example 1: Preparation of herbal extracts

The herbal medicine extract used in the present invention was provided by Hanpoong Pharmaceutical and complied with the herbal medicine raw material test regulations. As for the extraction method, after extracting from 30% alcohol, all the alcohol was removed using a vacuum concentrator, and then freeze-dried to separate the extracted powder. The obtained powder was dissolved in tertiary distilled water at 200 mg/ml and used.

Example 2: Culture of malignant glioblastoma cells

U87MG glioblastoma and HEK293T (Human embryonic kidney cells 293T) used in the present invention were pre-sale from the Korea Cell Line Bank (Seoul National University Cancer Research Institute) and used in the experiment. U87MG and HEK293T cells were prepared in DMEM (Dulbecco Modified Eagle Medium) with 10% FBS (Fetal Bovine Serum) and 1% antibiotic (P/S, Penicillin and Streptomycin) added to the cell culture solution at 37°C and 5%. The culture was maintained in a cell incubator in which CO ₂ was maintained.

Example 3: Inhibition of growth of malignant glioblastoma by loss of Axl

Since Axl is known to play a very important role in cancer cell growth and anticancer drug resistance in almost all carcinomas, Axl expression was lost to investigate the effect of loss of Axl expression on brain cancer cell growth.

More specifically, viruses expressing two different Axl shRNAs (#2, #4) were infected with U87MG and U373MG cells using lentiviral shRNA, thereby losing the expression of Axl. The infected U87MG and U373MG cells were inoculated into a 96-well plate in a number of 5×10 ⁴ after 5 days, and the cells were cultured for 48 hours, followed by MTT analysis.

1 μg of pLKO.1/shNS or pLKO.1/shAxl, 0.75 μg psPAX2 (packaging plasmid), and 0.25 μg pDM2.G (envelope plasmid) of 1 μg pLKO.1/shNS or pLKO.1/shAxl, 0.75 μg pDM2.G (envelope plasmid) were added to Lipofectamine 3000 reagent the next day after plating 1×10 ⁶ ml of HEK293T onto a ⁶ well plate. And delivered into cells through liposome-mediated introgression. At the 2nd and 3rd days after introgression, 2ml of cell culture solution was collected and filtered using a 0.4 μm syringe filter to infect cells.

After culturing the infected cells for 5 days, 200 μl of 0.5 mg/ml MTT reagent was dispensed and further cultured at 37° C. for 2 hours. After removing the MTT reagent, and dissolving all the formazan formed by dispensing 200 μl of DMSO into each well, the absorbance was measured at 540 nm using an ELISA reader.

As a result, as shown in Figure 1, compared to the control cells in the U87MG and U373MG cells in which Axl expression was lost, both shAxl #2 and shAxl #4 showed a cell growth rate of 50% or less.

Through this, it was found that Axl expression is a protein that plays a very important role in the growth of malignant glioblastoma.

Example 4: Cell signaling system change in malignant glioblastoma growth due to Axl loss

The expression of ERK, p38, JNK1, AKT, S6K and STAT3 was analyzed in order to examine the cell signaling pathway through which loss of Axl inhibits the growth of malignant glioblastoma.

More specifically, ERK, p38, JNK1, AKT, S6K and STAT3 are representative signaling systems closely related to cancer cell growth. Therefore, the degree of phosphorylation of each protein exhibiting the activity of ERK, p38, JNK1, AKT, S6K and STAT3 in U87MG and U373MG cells in which Axl was lost was confirmed through Western blotting.

Wash the U87MG and U373MG with PBS and RIPA buffer (150mM NaCl, 1% NP-40, 0.5% DOC, 0.1% SDS, 50mM Tris (pH8.0), 1mM EDTA, 1mM PMSF, 1mM NaF, 1mM Na3PO4, 1μg /mL aprotinin, leupeptin, pepstatin) was dissolved and the protein was separated by centrifugation at 13,000 xg for 20 minutes.

20 μg of the same amount of protein was separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then the separated protein was transferred to a nitrocellulose membrane (NC) using a Trans-blot unit. . The protein-transferred membrane was blocked with 5% fat-free skim milk powder (Tris buffered saline containing 0.1% Tween-20: TBS-T) for 1 hour at room temperature, and the primary antibody diluted to a concentration of 1:1000 was 4 It was reacted at °C for 16 hours. After washing in PBS, the secondary antibody was diluted 1:5,000 and reacted at room temperature for 1 hour. After the washing process, Pierce ECL Western Blotting Substrate was applied to the film, and the band was confirmed by reacting with the film in a dark room.

As a result, as shown in FIG. 2, it was found that only the activity of STAT3 was decreased by the loss of Axl in the U87MG and U373MG cells, and there was no change in other signaling systems.

Through this, it was confirmed that Axl loss inhibited the growth of glioblastoma by reducing the activity of STAT3.

Example 5: Screening for herbal medicines to reduce the amount of Axl protein expression

Through the above process, it was confirmed that the decrease in Axl expression of glioblastoma inhibited the growth of glioblastoma. Therefore, in order to find herbal medicines that reduce Axl expression, water extracts of 26 kinds of herbal medicines were treated with U87MG cell line, a representative malignant glioblastoma, and the amount of Axl expression was confirmed through western blotting.

More specifically, the herbal medicine water extract was treated for 24 hours at a concentration of 500 μg/ml, respectively. Extraction, cell culture, and Western blotting methods of herbal medicine are as described above.

As a result, as shown in FIG. 3, it was confirmed that the thickness of the protein band decreased when bokbunja, cheonhwabun, gold, or hwangbaek were treated.

Through this, it was confirmed that bokbunja, cheonhwabun, golden or hwangbaek can be used as herbal medicines that can treat glioblastoma by inhibiting and reducing the expression of Axl.

Example 6: Effect of Hwangbaek and Golden Extracts on Growth Inhibition of Malignant Glioblastoma

Among the herbal extracts for which Axl expression loss ability was verified, MTT analysis was performed by including the extract in the cell culture solution and culturing U87MG cells to see what role Hwangbaek or Golden extract has on the growth of malignant glioblastoma cells.

More specifically, after inoculation of U87MG cells in a 96-well plate at 5×10 ⁴ /ml number, the next day, Hwangbaek extract was treated at a concentration of 500 μg/ml for 48 hours, and golden extract was treated at a concentration of 20 to 1000 μg/ml. . The extraction method, cell culture method, and MTT analysis method of the herbal medicine used in the present invention are as described above.

As a result, as shown in FIG. 4, U87MG cells grown in a cell culture solution containing 500 μg/ml of Hwangbaek extract showed a cell growth rate of about 65% compared to the control without extract, as shown in FIG. The golden extract showed a cell growth rate of about 30% compared to the control at 1000 μg/ml.

Through this, it was confirmed that the golden or Hwangbaek extract has the ability to inhibit the expression of Axl in U87MG cells and the growth of glioblastoma.

Example 7: Changes in cell signaling system of glioblastoma by Hwangbaek and golden extracts

Through the above process, it was confirmed that the activity of the STAT3 signaling system decreased in the glioblastoma in which Axl was lost. It was confirmed through Western blotting that the Hwangbaek and Golden extracts inhibit the growth of cancer cells through which cell signaling pathways.

More specifically, after treating U87MG cells with a 500 μg/ml concentration of golden or yellow white extract for 48 hours, Western blotting was performed to determine the degree of phosphorylation of each protein showing the activities of ERK, p38, JNK1, AKT, S6K and STAT3. The western blotting method was confirmed and used as described above.

As a result, as shown in FIG. 6, when the Hwangbaek and Golden extracts at a concentration of 500 μg/ml were treated for 48 hours, there was no change in other signaling systems of U87MG cells, but it was confirmed that the activity of STAT3 was rapidly decreased.

Through this, it was confirmed that the Hwangbaek or Golden extract inhibited the growth of glioblastoma by reducing STAT3 activity, which was confirmed to exhibit similar effects to the inhibition of activity through viral infection.

In conclusion, the present invention presented a natural anticancer agent against glioblastoma by discovering an herbal medicine targeting Axl of glioblastoma. Based on the results of previous studies that Axl plays an important role in the growth of cancer cells and that treatment with Axl inhibitors reduces the growth of cancer cells, we examined how loss of Axl affects the growth of glioblastoma. The loss of Axl caused by the inhibition of the growth of U87MG and U373MG by less than 50%, and through this, it was confirmed that Axl plays an important role in the growth of glioblastoma.

In addition, Hwangbaek, Golden, or Bokbunja extracts suggested the possibility of treating glioblastoma by reducing the expression of Axl protein, and among them, Hwangbaek or Golden extract inhibited the growth of cancer cells in the cell line and reduced the expression of Axl protein. It was found that inhibiting the activity of STAT3 ultimately inhibited the growth of glioblastoma.

Through this, it was suggested that Hwangbaek, Golden, or Bokbunja extracts could be effectively used for the treatment of glioblastoma, as well as that Axl could be a new effective target for the treatment of glioblastoma.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features thereof. In this regard, the embodiments described above are illustrative in all respects and should be understood as non-limiting. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the claims to be described later rather than the above detailed description, and equivalent concepts thereof.

Claims (8)

A pharmaceutical composition for the prevention or treatment of glioblastoma, comprising as an active ingredient any one or more extracts selected from the group consisting of golden, hwangbaek and bokbunja or a fraction thereof.
The pharmaceutical composition of claim 1, wherein the extract is prepared by extraction with a solvent selected from the group consisting of water, an alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof.
The pharmaceutical composition of claim 1, wherein the extract is a hot water extract.
The pharmaceutical composition of claim 1, wherein the extract contains 0.0001 to 10% by weight based on the total composition.
The pharmaceutical composition according to claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent.
A method for preventing or treating glioblastoma, comprising administering the composition of any one of claims 1 to 5 to an individual other than a human.
A food composition for preventing or improving glioblastoma, comprising any one or more extracts selected from the group consisting of golden, hwangbaek and bokbunja or a fraction thereof as an active ingredient.
A feed composition for preventing or improving glioblastoma, comprising any one or more extracts selected from the group consisting of golden, hwangbaek and bokbunja or a fraction thereof as an active ingredient.

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