KR20220054781A - 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

A composition for preventing or treating glioblastoma comprising an herbal medicine extract {Pharmaceutical composition for preventing or treating glioblastoma comprising Phellodendron amurense Ruprecht or Scutellaria baicalensis}

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

GBM (glioblastoma multiforme) refers to a glioma with the highest malignancy 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, radiation therapy, and chemotherapy using temozolomide, but the average survival rate is only 14 months, so a more effective treatment is needed.

Axl receptor tyrosine kinase belongs to the TAM (Tyro3-Axl-Mertk) receptor group, and has recently been attracting 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, and a single-pass transmembrane domain and a tyrosine phosphorylation domain inside the cell. Growth arrest-specific gene6 (GAS6) and Protein S (PROS1) are known as ligands that bind to the Axl receptor and induce activation.

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

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 drugs targeting Axl have been developed and clinical trials are being actively conducted. It was found that Axl inhibition was effective as a way to overcome the fact that Axl inhibitors induce cancer cell death and anticancer drug resistance to PARP inhibitors and PI3K inhibitors.

As such, the importance of Axl in cancer research has been greatly emphasized through research results over the past 10 years. Volume 198, 1 April 2018, Pages 99-111), the development of Axl-targeted natural anticancer drugs is non-existent.

Under this background, the present inventors made intensive research efforts to discover oriental medicines that inhibit the growth of glioblastoma by inhibiting the expression of Axl by targeting Axl. The present invention was completed.

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

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

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

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 or fractions thereof selected from the group consisting of gold, hwangbaek and bokbunja as an active ingredient.

As used herein, the term "golden (Scutellaria baicalensis)" refers to a drug made from the roots of gold (Scutellaria baicalensis GEORGE), a perennial herbaceous plant belonging to the family Lamiaceae. The gold is generally grown on the grass in the mountainous area, and several generations come out and grow into leaves, hairy, and branched. The root is conical and the flesh is yellow. In oriental medicine, the root is used as an antipyretic, diuretic, diuretic, edema, and anti-inflammatory agent. On the other hand, baicalin, a kind of flavonoid contained in the gold, is known to have antioxidant, anticancer, and anti-inflammatory effects.

As used herein, the term “hwangbaek (Phellodendron amurense Ruprecht)” refers to the bark of a phlegm tree used as a medicine, and is also called hwanggyeongpi. It is a deciduous broad-leaved arboreous tree that grows to about 10m in height. Cork is developed on the bark, and the inner bark is yellow. In Korea, it is distributed evenly in all regions except Jeju and Jeollanam-do, and it grows in arboreal forests or mountain valleys. The hwangbaek is used as a medicine by peeling the bark from the tree trunk before and after the summer solstice, removing or cutting the rough skin (粗皮: rough bark) and drying it in the sun. It is known that the yellow white has a blood sugar lowering action, inhibits the growth of pneumococci, tuberculosis pneumococcus, staphylococcus, and the like and the proliferation of tumor cells, and has a sterilizing action.

As used herein, the term "Rubus coreanus" belongs to the Rosaceae family and is known to be native to China, and Jeju Island and southern regions are known as major production areas in Korea. Bokbunja is harvested from dark red fruits in early summer and used for food. Since ancient times, bokbunja has been known as a valuable medicinal agent in oriental medicine as a tonic, stimulant, and liver-protecting summary.

As used herein, the term “extract” refers to a substance obtained by extracting a certain substance, specifically, an extract obtained by the extraction treatment of the present invention, a diluted or concentrated solution of the extract, and a dried product obtained by drying the extract , It includes extracts of all formulations that can be formed using the extract itself and the extract, such as a prepared or purified product of the extract, or a mixture thereof.

The regular extract may be a single extract of gold, hwangbaek, or bokbunja, and may be two or more complex extracts selected from the group consisting of gold, hwangbaek and bokbunja, but is not limited thereto. More specifically, golden and yellow-white extracts; hwangbaek and bokbunja extract; golden and raspberry extract; or extracts of golden, yellow, and bokbunja; can be

A 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 may 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 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” refers to a result obtained by performing fractionation in order to separate a specific component or a specific group of components from a mixture including various components.

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

In the present invention, the type 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 water, distilled water, a polar solvent such as alcohol; and non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of two or more. In the case of using alcohol in the fractionation solvent, preferably C1 to C4 alcohol may be used.

As used herein, the term “prevention” refers to any action that inhibits or delays the growth of glioblastoma by administration of the pharmaceutical composition according to the present invention.

As used herein, the term “treatment” refers to any action in which the symptoms of a suspected and diseased individual of glioblastoma are improved or beneficially changed by administration of the pharmaceutical composition.

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

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

More specifically, carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, 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 (poly-L-lactic acid), mineral oil, and the like.

The pharmaceutical composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories and sterile injection solutions according to conventional methods, respectively. The carrier may include various amorphous carriers, microspheres, nanofibers, and the like.

In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used.

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

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

Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

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

In this case, the description of the “pharmaceutical composition”, “prevention” and “treatment” is the same as described above.

Since the pharmaceutical composition of the present invention exhibits a preventive or therapeutic effect on glioblastoma, the method of the present invention comprising administering the same to an individual can be usefully utilized for the prevention or treatment of glioblastoma.

As used herein, the term “administration” means introducing the composition to a subject by an appropriate method.

As used herein, the term "individual" refers to all animals, such as mice, mice, and livestock, including humans, that have or can develop glioblastoma, and specifically, 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 or fractions thereof selected from the group consisting of gold, hwangbaek and bokbunja.

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

As used herein, the term “improvement” refers to any action that at least reduces a parameter, eg, the severity of a symptom, associated with a condition to be treated by administration of the composition.

Since the extract of gold, hwangbaek or bokbunja according to the present invention exhibits an excellent glioblastoma inhibitory effect, it may be included in a food composition for the purpose of preventing or improving glioblastoma, and since the food composition can be ingested on a daily basis, glioblastoma A high effect can be expected for the prevention or improvement of

As used herein, 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. include all foods in the usual sense, and as long as it can contain gold or yellow white of the present invention, it is not limited thereto.

As used herein, the term “health functional food” refers to food manufactured and processed using raw materials or ingredients useful for the human body in accordance with Act No. 6727 of the Health Functional Food Act, and 'functionality' refers to the structure of the human body. And it means to obtain a useful effect for health use, such as regulating nutrients for function or physiological action. On the other hand, health food means food that has an active health maintenance or promotion effect compared to general food, and health supplement means food for the purpose of health supplementation. In some cases, the terms health functional food, health food and health supplement food can be mixed.

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

The food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art. Specifically, the food composition may further include a physiologically acceptable carrier, the type of carrier is not particularly limited and any carrier commonly used in the art may be used. In addition, the food composition contains food additives such as preservatives, disinfectants, antioxidants, colorants, coloring agents, bleaching agents, seasonings, sweeteners, flavoring agents, expanding agents, strengthening agents, emulsifying agents, thickeners, filming agents, gum base agents, foam inhibitors, solvents, and improving agents. may 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 composition for food of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage of not having side effects that may occur during long-term administration of drugs using food as a raw material, and is excellent in portability, so the present invention Foods of can be ingested as an adjuvant to enhance the effect of prevention or improvement of glioblastoma.

Gold, hwangbaek or bokbunja of the present invention may be included in various weight % in the food composition if it can exhibit the effect of preventing or improving glioblastoma. Specifically, it may be included in an amount of 0.00001 to 100% by weight or 0.01 to 80% by weight relative to the total weight of the food composition, but is not limited thereto. When ingested for a long period of time for health and hygiene purposes, it may contain an amount below the above range, 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 or fractions thereof selected from the group consisting of gold, hwangbaek, and bokbunja.

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

Gold, Hwangbaek or bokbunja according to the present invention exhibits an excellent therapeutic effect on glioblastoma, and thus may be included in a feed composition for the purpose of preventing or improving glioblastoma. A high effect can be expected for the prevention or improvement of blastoma.

As used herein, the term "feed" means any natural or artificial diet, meal, etc., or a component of said meal, intended for or suitable for being eaten, consumed, and digested by an animal.

The type of feed is not particularly limited, and feed commonly used in the art may be used. Non-limiting examples of the feed include plant feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, gourds or grain by-products; and animal feeds such as proteins, inorganic materials, oils and fats, minerals, oils and fats, single cell proteins, zooplankton, or food. These may be used alone or in mixture of two or more.

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

1 is a graph comparing the effect of Axl on glioblastoma growth by loss of Axl gene through shRNA in U87MG and U373MG cells, which are glioblastomas, through MTT analysis.
2 is a graph analyzed by Western blotting to determine what mechanism was inhibited to inhibit the growth of glioblastoma in glioblastoma U87MG and U373MG in which the Axl gene was lost.
3 is a graph showing the results of comparing the expression levels of Axl and GAPDH through western blotting after 24 hours of treatment with water extracts of 26 kinds of herbal medicines in U87MG, a glioblastoma, in order to find oriental medicines capable of inhibiting Axl; am.
4 is a graph showing the results of fertilizing the growth of the cells by treating the Hwangbaek extract, which showed the effect of inhibiting Axl, to U87MG, which is a glioblastoma, for 48 hours.
5 is a graph showing the results of fertilizing the growth of the cells by treating the golden extract showing the effect of inhibiting Axl to U87MG, a glioblastoma, for 48 hours.
FIG. 6 is a graph analyzed by Western blotting to see if gold and yellow white, which had the effect of inhibiting the growth of glioblastoma, inhibited the growth of glioblastoma by inhibiting any mechanism.

Hereinafter, the present invention will be described in more detail by way of Examples. However, the following examples are only for illustrating the present invention, 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. In the extraction method, after extracting from 30% alcohol, all of the alcohol was removed using a vacuum concentrator, and then the extracted powder was separated by freeze-drying. The obtained powder was dissolved in tertiary distilled water at 200 mg/ml and used.

Example 2: Glioblastoma malignant cell culture

U87MG glioblastoma and HEK293T (Human embryonic kidney cells 293T) used in the present invention were purchased from the Korea Cell Line Bank (Seoul National University Cancer Institute) and used in the experiment. U87MG and HEK293T cells were cultured using a cell culture medium supplemented with 10% FBS (Fetal Bovine Serum) and 1% antibiotics (P/S, Penicillin and Streptomycin) in DMEM (Dulbecco Modified Eagle Medium) 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 glioblastoma malignant by Axl loss

Since Axl is known to play a very important role in cancer cell growth and resistance to anticancer drugs in almost all carcinomas, in order to examine the effect of loss of Axl expression on brain cancer cell growth, it was confirmed by loss of Axl expression.

More specifically, viruses expressing two different Axl shRNAs (#2, #4) were infected with lentiviral shRNA to U87MG and U373MG cells to abolish the expression of Axl. Infected U87MG and U373MG cells were inoculated into 96-well plates in the number of 5x10 ⁴ after 5 days, and after culturing the cells for 48 hours, MTT analysis was performed.

The next day after plating 1x10 ⁶ ml of HEK293T on a 6 well plate, 1 μg of pLKO.1/shNS or pLKO.1/shAxl, 0.75 μg psPAX2 (packaging plasmid), and 0.25 μg pDM2.G (envelope plasmid) were mixed with Lipofectamine 3000 reagent. It was delivered into cells through use, liposome-mediated introgression. At 2 and 3 days after introgression, 2 ml of the cell culture solution was collected, and filtered using a 0.4 μm syringe filter to infect the 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 200 μl of DMSO into each well to dissolve all of the formazan, absorbance was measured at 540 nm using an ELISA reader.

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

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

Example 4: Changes in Cell Signaling System in Growth of Glioblastoma Malignant by Axl Loss

Expression of ERK, p38, JNK1, AKT, S6K and STAT3 was analyzed to examine which cell signaling pathway Axl loss inhibits the growth of glioblastoma malignancy.

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

The U87MG and U373MG were washed with PBS, and RIPA buffer (150 mM NaCl, 1% NP-40, 0.5% DOC, 0.1% SDS, 50 mM Tris (pH8.0), 1 mM EDTA, 1 mM PMSF, 1 mM NaF, 1 mM Na3PO4, 1 μg /mL aprotinin, leupeptin, pepstatin) was dissolved and the protein was separated by centrifugation at 13,000×g 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% nonfat skim milk powder (Tris buffered saline containing 0.1% Tween-20: TBS-T) for 1 hour at room temperature, and the primary antibody diluted at a concentration of 1:1000 was 4 It was reacted at ℃ 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 membrane, and the band was confirmed by reacting it 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 reduced by Axl loss in U87MG and U373MG cells, and there was no change in other signaling systems.

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

Example 5: Screening of herbal medicines that reduce the expression level of Axl protein

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

More specifically, the herbal medicine water extracts were each treated at a concentration of 500 μg/ml for 24 hours. Extraction, cell culture, and Western blotting methods of herbal preparations are the same as described above.

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

Through this, it was confirmed that bokbunja, cheonhwabun, golden or yellow fern can be a herbal medicine that can treat glioblastoma by inhibiting and reducing the expression of Axl.

Example 6: Glioblastoma malignant growth inhibitory effect of Hwangbaek and Golden extract

MTT analysis was performed by including the extract in a cell culture medium and culturing U87MG cells in order to examine the role of Hwangbaek or Golden extract among herbal extracts whose Axl expression loss ability has been verified on the growth of glioblastoma malignant cells.

More specifically, U87MG cells were inoculated in a 96-well plate at a number of 5x10 ⁴ /ml, and the next day, the Hwangbaek extract was treated at a concentration of 500 μg/ml for 48 hours, and the golden extract was treated at a concentration of 20 to 1000 μg/ml. . The extraction method of the herbal preparation used in the present invention, the cell culture method, and the MTT analysis method are the same as described above.

As a result, as shown in FIG. 4, U87MG cells grown in a cell culture medium containing 500 μg/ml of Hwangbaek extract showed a cell growth rate of about 65% compared to the control containing no extract, and as shown in FIG. 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 had the ability to inhibit the expression of Axl in U87MG cells and to inhibit the growth of glioblastoma.

Example 7: Changes in the cell signaling system of glioblastoma by Hwangbaek and Golden extract

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

More specifically, after treating gold or yellow white extract at a concentration of 500 μg/ml in U87MG cells for 48 hours, Western blotting was performed to determine the phosphorylation level of each protein indicating the activity of ERK, p38, JNK1, AKT, S6K and STAT3. The western blotting method used was as described above.

As a result, as shown in FIG. 6 , when 500 μg/ml of Hwangbaek and gold extract was 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 Hwangbaek or Golden extract inhibits the growth of glioblastoma by reducing STAT3 activity, which has a similar effect to inhibition of activity through virus infection.

In conclusion, the present invention discovered a herbal medicine targeting Axl of glioblastoma and presented a natural anticancer agent for glioblastoma. Based on the previous study that Axl plays an important role in the growth of cancer cells and treatment with Axl inhibitors reduces cancer cell growth, we looked at how the loss of Axl affects the growth of glioblastoma. As a result, lentiviral shRNA The loss of Axl by induced inhibition inhibited the growth of U87MG and U373MG by 50% or less, confirming that Axl also plays an important role in the growth of glioblastoma.

In addition, Hwangbaek, gold or bokbunja extract reduced the expression of Axl protein, suggesting the possibility of treating glioblastoma. Among them, Hwangbaek or Golden extract inhibited the growth of cancer cells in cell lines, It was found that by inhibiting the activity of STAT3, it ultimately inhibits the growth of glioblastoma.

This suggested that Hwangbaek, gold or bokbunja extract could be effectively used for the treatment of glioblastoma, and that Axl could be a novel and efficient 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 may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the following claims and their equivalents.

Claims (7)

A pharmaceutical composition for the prevention or treatment of glioblastoma, comprising an alcohol mixture extract of gold and bokbunja as an active ingredient.
The pharmaceutical composition of claim 1, wherein the extract comprises 0.0001 to 10% by weight of the total composition.
The pharmaceutical composition of 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 3 to a subject other than a human.
A food composition for the prevention or improvement of glioblastoma, comprising an alcohol mixture extract of gold and bokbunja as an active ingredient.
A feed composition for preventing or improving glioblastoma, comprising an alcohol mixed extract of gold and bokbunja as an active ingredient.
A composition for inhibiting Axl expression in vitro, comprising an alcohol mixture extract of gold and bokbunja as an active ingredient.

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