KR101523434B1 - A composition comprising Julbernardia globiflora extracts having anti-cancer activity - Google Patents

Abstract

The present invention relates to a process for the preparation of < RTI ID = 0.0 > Julbernardia < globiflora extract as an active ingredient, and a food composition for preventing or ameliorating cancer. The composition comprising the extract of Zulberna diaglobifolia of the present invention shows apoptosis of cancer cells without showing toxicity to normal cells and shows a cancer cell killing effect and thus can be usefully used as an anti-cancer composition, Or as a pharmaceutical or food composition for treatment.

Description

[0001] The present invention relates to an anticancer composition comprising Julbernardia globuliflora extract as an active ingredient,

The present invention relates to a process for the preparation of < RTI ID = 0.0 > Julbernardia < globiflora extract as an active ingredient, and a food composition for preventing or ameliorating cancer.

Cancer is one of the major causes of death in Korea. There have been many studies to conquer cancer. However, it is an incurable disease that can not be regulated. In cancer, cells divide and grow uncontrollably to form malignant tumors and invade adjacent tissues of the body. In addition, cancer can migrate farther through lymphatic or blood flow as well as metastasis to adjacent tissues. Existing therapies for cancer include surgery, chemotherapy, and radiation therapy. Among these, chemotherapy using chemotherapeutic agents is now widely used for cancer treatment and is one of relatively well established treatment methods.

The term 'anticancer agent' refers to chemotherapeutic agents other than surgery and radiation therapy among the methods used to treat malignant tumors, and most of them are substances that inhibit the synthesis of nucleic acids and exhibit anticancer activity. Chemotherapeutic agents are largely classified into antimetabolites, alkylating agents, antimitotic drugs, and hormones. Methotrexate, 6-mercaptopurine, 6-thioguanine, 5-fluorouracil, and Cytarabine are examples of metabolic antagonists that inhibit the metabolic processes required for cancer cell proliferation. Examples of the alkylating agent that introduces an alkyl group into the DNA guanine to modify the DNA structure and cleave the chain to exhibit an anticancer effect include chlorambucil, cyclophosphamide, ethylenimine compound, thiotepa, (busulfan), a nitrosourea compound (carmustine), and a triazine compound (dacarbazine). As mitotic timing specificity drugs, mitotic inhibitors that inhibit mitosis by inhibiting mitosis by inhibiting mitosis include anticancer drugs such as actinomycin D, doxorubicin, bleomycin, mitomycin, vincristine, vinblastine Plant alkaloids, taxoids that are mitotic inhibitors including taxane rings, and the like. In addition, hormones such as corticosteroids or progesterone and platinum-containing compounds such as cisplatin are used as anticancer drugs.

Such anticancer agents interfere with the metabolism pathway of cancer cells and interfere with DNA replication, transcription, and translation by direct interaction with DNA, interfere with the synthesis of nucleic acid precursors, and inhibit cell division, thereby exhibiting toxicity to cells. However, substances generally used as anticancer drugs have considerable toxicity, and they do not selectively remove only cancer cells and cause fatal damage to normal cells, resulting in hemopoiesis such as leukocytes, platelets and red blood cells due to bone marrow destruction; Hair loss symptoms due to hair follicle cell destruction; Side effects of ovaries and testicles cause menstrual irregularities and male infertility; Side effects from digestive mucous membrane destruction include stomatitis, nausea and vomiting and food swallowing disorders and digestive disorders; Diarrhea symptoms; Renal toxicity by tubular necrosis; Peripheral neuritis and weakness caused by nervous system disorders; Vascular disorders such as vascular pain and rash; Skin and nail discoloration.

Therefore, it is urgently required to develop a chemotherapeutic agent having low toxicity and effective to prevent the development of cancer, as well as research to raise the therapeutic effect while minimizing side effects of the anticancer agent and treatment of cancer after the cancer. In addition, the biggest problem of chemotherapeutic agents is drug resistance, which is the main cause of failure in the end despite the initial successful response by the anticancer drug. Therefore, It is becoming important.

Accordingly, the inventors of the present invention have made efforts to search for natural products having anticancer effects and less side effects, and as a result, it has been confirmed that an extract of Zulberna diaglobifolia, a kind of foreign bioactive plants, can kill cancer cells effectively, and the present invention has been completed .

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cancer comprising an extract of Julbernardia globiflora as an active ingredient.

Another object of the present invention is to provide a method for the treatment of < RTI ID = And to provide a food composition for preventing or ameliorating cancer comprising an extract as an active ingredient.

In one aspect of the present invention, the present invention provides a pharmaceutical composition comprising Julbernardia < RTI ID = 0.0 > globiflora ) extract as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating cancer.

The term " julbernardia " globiflora "is a genus Julbernardia plant of the family Fabaceae or Leguminosae belonging to the Angiosperms . It is widely distributed at medium altitudes in the Miombo forest area to the southern and eastern parts of the equator forest area of the Congo Basin. It is an existing tropical African tree, also called Mnondo. It exists over a large area and is ecologically important and less It dominates as co-dominant in various forms of woodland, such as growing well in the drier form of competitive deciduous forests. It is far from the coast except the southern limit. I prefer hot, dry rest and rainy summers. It grows well in places where daily variability is noticeable. It is a medium-sized tree with tens of meters of deciduousness. Preferably, the Zulberna diaglubi flora may be Julbernardia globiflora Troupin. On the other hand, the growth characteristics of the above-mentioned zebernadiolabi flora are known, but the activity thereof, particularly the anticancer activity has not been known at all.

The term "extract " of the present invention means a preparation which is obtained by squeezing a herbal medicine with an appropriate leaching solution and concentrating by evaporating the leaching solution, and is not limited thereto. The extract, the diluted solution, A dried product obtained by drying, a controlled preparation thereof, or a purified product thereof. The sulbated diaglobiophyla trough pin Extracts can be prepared using common extraction, isolation and purification methods known in the art. The extraction method may be, but not limited to, hot water extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.

According to a specific embodiment of the present invention, the Zulberna diaglobrifloratrowin extract of the present invention is prepared by crushing a certain amount of Zulberna diaglobiprophatrachin powder into powder and extracting it with 5 to 10 times of a solvent. As the solvent, an organic solvent may be used, and the organic solvent may be water, C1 to C4 lower alcohol or a mixture thereof, preferably methanol. Further, the extraction can be preferably carried out at a temperature of 35 to 55 ° C, more preferably at 45 ° C. As described above, the extraction method can be carried out by using any method known in the art without limitation, but it can be preferably extracted by ultrasonication. The extraction was repeated several times, and the extracted sample was filtered, concentrated and lyophilized.

In the present invention, the term "prevention" means any action which inhibits or delays the development, spread and recurrence of cancer by the administration of the pharmaceutical composition according to the present invention, and "treatment" Suspicion of cancer and all the behaviors that alleviate or ameliorate symptoms of the onset individuals.

The cancer which can be prevented or treated by the composition of the present invention is a cancer selected from the group consisting of cervical cancer, lung cancer, non-small cell lung cancer, pancreatic cancer, colon cancer, liver cancer, colon cancer, bone cancer, skin cancer, head cancer, Hodgkin's disease, Esophageal cancer, Small intestine cancer, Endocrine cancer, Endometrial carcinoma, Endometrial carcinoma, Endometrial carcinoma, Endometrial carcinoma, Vaginal cancer, Hodgkin's disease, Uterine cancer, Ovarian cancer, Rectal cancer, Brain tumor, Bladder cancer, Renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, renal cell carcinoma, renal cell carcinoma, renal cell carcinoma, kidney cancer, pancreatic cancer, thyroid cancer, pituitary cancer, adrenal cancer, soft tissue sarcoma, , Spinal cord tumor, brainstem glioma, and pituitary adenoma. Preferably lung cancer, colorectal cancer, or liver cancer, and more preferably, it can be, but is not limited to, colorectal cancer or liver cancer.

Preferably, the prevention or treatment of the cancer can be accomplished by the apoptosis of cancer cells induced by the urea diaglabi flora extract according to the present invention. The above-mentioned "apoptosis" is a kind of programmed cell death (PCD), in which abnormal cells, damaged cells and aged cells are killed and killed by the biological program inputted into our body, . Such apoptosis is distinguished from cell death caused by stimulation such as burns, scabs, and poisons, such as necrosis, which is the death or accidental death of a cell. That is, apoptosis is an active death of a cell that is regulated by a protein expressed from a specific gene. While apoptosis occurs chaotically, apoptosis occurs in an orderly fashion in a short time. Specifically, apoptosis begins with the contraction of the cell, creating a gap between the adjacent cells, and DNA, which is a genetic material in the nucleus of the cell in the process of dying, is regularly cleaved into fragments. Ultimately, the entire cell called the apoptotic body is also broken up, and it is killed by other cells around it. It is involved in the formation of the human body during the early development of the human body. In adults, it is responsible for eliminating normal cells when they become senescent, or removing cells with abnormalities. The apoptosis is distinguished from PCD in that it also occurs in cases other than PCD, and is related to cell death due to cancer cell death, viral infection, drug, and radiation.

According to a specific embodiment of the present invention, the composition of the present invention, such as, for example, Methanol extracts showed no cytotoxicity against normal cells (3T3-L1) but showed killing effect on cancer cell lines (A549, HT29 and HepG2). Especially, it was found that colon cancer cells (HT29) and hepatocyte (Fig. 1). In addition, it was confirmed that the treatment of the composition induces apoptosis and induces apoptosis. In particular, in the case of colorectal cancer and hepatic cancer cells, apoptosis is effectively induced upon treatment with Zebernadagloviflora extract, (Figures 2 and 3). Further, by confirming the expression of proteins involved in the apoptosis of cancer cells, it can be confirmed once again that the composition of the present invention induces apoptosis of cancer cells through the external pathway through the death receptor and the internal pathway in the mitochondria (Fig. 4).

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" of the present invention means that it exhibits properties that are not toxic to the cells or humans exposed to the composition. Such carriers may be used without limitation as long as they are known in the art such as buffers, preservatives, wetting agents, solubilizers, isotonic agents, stabilizers, bases, excipients and lubricants.

In addition, the pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method have. Furthermore, it can be used in the form of an external preparation for skin in the form of ointments, lotions, spray agents, patches, creams, powders, suspensions, gels or gels. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate ), Sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension, liquid solutions, emulsions and syrups have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

Meanwhile, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount " of the present invention means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, The type of cancer, the severity, the activity of the drug, the sensitivity to the drug, the method of administration, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including the drugs used concurrently or concurrently and other factors well known in the medical arts . The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the extract contained in the composition of the present invention may vary depending on the age, sex, and body weight of the patient. Generally, the extract of Zulberna diaglobifolia of the present invention is administered per kg of body weight per day Preferably 1 to 10 mg / kg body weight per day, more preferably 1 to 5 mg / day, or one to three divided doses per day. However, the dosage may not be limited in any way because it may be increased or decreased depending on route of administration, severity of disease, sex, weight, age, and the like.

The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

In another aspect, the present invention provides a method of preventing or treating cancer, comprising administering a diabetic strain to a subject in need thereof.

The term "individual" of the present invention means a mammal such as a monkey, a cow, a horse, a sheep, a pig, a chicken, a turkey, a quail, a cat, a dog, a mouse, a rat, a rabbit or a guinea pig , And the diseases can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to an individual. The pharmaceutical composition of the present invention can be administered in parallel with existing therapeutic agents.

The term "administering" of the present invention means providing the patient with the desired substance in any suitable manner, and the administration route of the composition of the present invention may be administered through any conventional route so long as it can reach the target tissue have. But are not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrathecal, rectal. In addition, the pharmaceutical composition of the present invention may be administered by any device capable of moving the active substance to the target cell. The preferred modes of administration and formulations are intravenous, subcutaneous, intradermal, intramuscular, and drip injections. The injectable solution may be a non-aqueous solvent such as an aqueous solvent such as a physiological saline solution or a ring gel solution, a vegetable oil, a higher fatty acid ester (e.g., oleic acid), an alcohol (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.) (For example, ascorbic acid, sodium hydrogen sulfite, sodium pyrophosphate, BHA, tocopherol, EDTA and the like), an emulsifier, a buffer for pH control, a microbial growth inhibitor And a pharmaceutical carrier such as a preservative (e.g., mercury nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

The term "therapeutically effective amount " used in connection with the active ingredient in the present invention means the amount of the pharmaceutically acceptable salt of the urea dianthroblofura extract effective to prevent or treat the subject disease.

The pharmaceutical composition of the present invention may further include a known anticancer agent in addition to the effective amount of jwert's diaglabi flora extract, and may be used in combination with other therapies known for the treatment of these diseases. Other treatments include, but are not limited to, chemotherapy, radiation therapy, hormone therapy, bone marrow transplantation, stem cell replacement therapy, other biological therapies, immunotherapy, and the like.

Examples of anticancer agents that can be included in the pharmaceutical composition of the present invention include DNA alkylating agents such as mechloethamine, chlorambucil, phenylalanine, mustard, cyclophosphate, Cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin cisplatin and carboplatin; The anticancer antibiotics include dactinomycin (actinomycin D), doxorubicin (adriamycin), daunorubicin, idarubicin, mitoxantrone, Plicamycin, mitomycin C, and bleomycin; And plant alkaloids such as vincristine, vinblastine, paclitaxel, docetaxel, etoposide, teniposide, topotecan, And iridotecan, but are not limited thereto.

In another aspect, the present invention relates to a method of treating or preventing diabetes mellitus, A food composition for preventing or ameliorating cancer comprising an extract as an active ingredient. When the composition of the present invention is used as a food composition, it may be added as it is or may be used in combination with other food or food ingredients, and may be suitably used according to a conventional method. The composition may contain a food-acceptable food-aid additive in addition to the active ingredient, and the amount of the active ingredient to be mixed may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment).

As used herein, the term "food-aid additive " refers to a component that can be added to foods in a supplementary manner, and is appropriately selected and used by those skilled in the art as added to produce health functional foods of each formulation. Examples of food-aid additives include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, , a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated drink. However, the types of the food auxiliary additives of the present invention are not limited by these examples.

A health functional food may be included in the food composition of the present invention. The term "health functional food " as used in the present invention refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and rings using raw materials and components having useful functions in the human body. Here, 'functional' refers to the structure and function of the human body to obtain nutritional effects and obtain useful effects for health use such as physiological action. The health functional food of the present invention can be prepared by a method commonly used in the art and can be prepared by adding raw materials and ingredients that are conventionally added in the art. In addition, the formulations of the above health functional foods may also be manufactured without limitations as long as they are acceptable as health functional foods. The composition for food of the present invention can be manufactured in various forms, and unlike general pharmaceuticals, it has the advantage that there is no side effect that may occur when a drug is used for a long period of time, and is excellent in portability, Can be ingested as an adjuvant to enhance the effectiveness of anticancer drugs.

There is no limitation on the kind of health food to which the composition of the present invention can be used. In addition, the composition comprising the urea diaglabi flora extract of the present invention as an active ingredient may be prepared by mixing other suitable auxiliary ingredients and known additives, which may be contained in health functional foods, according to the selection of a person skilled in the art. Examples of foods that can be added include dairy products, such as meat, sausage, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Vitamin complex, and the like, and can be prepared by adding to the juice, tea, jelly, and juice prepared from the extract of the present invention as a main component.

The diabetic glioblastoma extract of the present invention can be used as a pharmaceutical composition or a food composition due to natural plant as a raw material, so that it can be safely contained in pharmaceutical composition and health functional food Lt; / RTI >

The composition comprising the extract of Zulberna diaglobifolia of the present invention shows apoptosis of cancer cells without showing toxicity to normal cells and shows a cancer cell killing effect and thus can be usefully used as an anti-cancer composition, Or as a pharmaceutical or food composition for treatment.

FIG. 1 is a graph showing the results of a methanol extract of Julbernardia globiflora ; MEJG) on cancer cell death.
FIG. 2 is a graph showing the cell death-inducing effect of MEJG according to the present invention on colon cancer cell HT29. (A) is a graph showing the results of flow cytometry on cells stained with FITC-Annexin V and PI (propidium iodide) after MEJG treatment, and (B) shows a change in cell survival rate according to the treatment concentration of the extract .
FIG. 3 is a graph showing the induction of cell death by MEJG according to the present invention on HepG2 cell line HepG2. (A) is a graph showing the results of flow cytometry on cells stained with FITC-Annexin V and PI (propidium iodide) after MEJG treatment, and (B) shows a change in cell survival rate according to the treatment concentration of the extract .
FIG. 4 is a graph showing the ability of MEJG according to the present invention to regulate apoptosis-related protein expression in colon cancer cell HT29. FIG.

Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example  One: Zuernardia Grobbi Flora  Trough pin ( Julbernardia globiflora Preparation of Troupin extract

As a foreign biologically active plant, Julbernardia ( USA) globiflora Troupin) was distributed through the Korea Plant Extract Bank of the Korea Biotechnology Research Center. The plant samples were put into 99.9% methanol and sonicated at 45 ° C with a sonicator to extract. The extract was concentrated, lyophilized and stored. In the experiment, the lyophilized sample was dissolved in dimethyl sulfoxide (DMSO).

Example  2: Culture of cell line

The methanol extract of Julbernardia < RTI ID = 0.0 > globiflora ; MEJG) and to confirm the cancer cell killing effect, the cell line 3T3-L1 and the cancer cell line A549, the colon cancer cell line HT29 and the hepatoma cell line HepG2 were purchased from the American Type Culture Collection (ATCC) Respectively. A549, HT29 and HepG2 were cultured in DMEM (Dulbecco's modified Eagle medium) medium containing 10% fetal bovine serum (FBS) and penicillin / streptomycin, and 3T3-L1 was cultured in 10% calf And cultured in DMEM medium containing serum (bovine calf serum; BCS) and penicillin / streptomycin.

Example  3: Zuernardia Grobbi Flora Trough pin  Analysis of cancer cell killing effect of extract

The safety of the MEJG prepared in Example 1 on normal cells and the specific killing effect on cancer cells were confirmed. Cancer cell lines and normal cell lines were treated with various concentrations (0 to 200 占 퐂 / ml) of MEJG after being adhered and cultured in a 24-well tissue culture dish at a rate of 1 x 10 ⁴ to 3 x 10 ⁴ cells per well, After incubation, cell death induction was analyzed by WST assay. The results are shown in FIG. 1 and Table 1.

As shown in Fig. 1, cancer cells (A549, HT29, and HepG2) do not show toxicity to the normal cell line (3T3-L1) And it was confirmed that cancer cell specific killing effect was exerted on both colorectal cancer cells and liver cancer cells. In particular, it showed excellent effects on colon cancer and liver cancer cells. The concentrations indicating 50% killing effect for each cancer cell line were calculated and are shown in Table 1.

Methanol extract IC ₅₀ ([mu] g / ml) A549 HT29 HepG2 MEJG 277.1 202.2 202.0

Example  4: Extract of Zulberna diaglobula floratrin Apoptosis ( apoptosis ) Induction ability  analysis

In order to elucidate the mechanism of the anticancer activity of the MEJG prepared in Example 1, the ability of the extract to induce apoptosis was confirmed. Particularly, when apoptosis is induced, phosphatidylserine, a component of the cell membrane, is exposed to the outside of the cell from inside the cell. Thus, the degree of apoptosis was confirmed using Annexin V, which specifically binds to phosphatidylserine. Specifically, cancer cells were cultured in a 6-well tissue culture dish at a concentration of 1 × 10 ⁵ to 3 × 10 ⁵ cells / well, and the cells were cultured for 48 hours at various concentrations of MEJG. V & Dead Cell Kit (Merck Millipore, Darmstadt, Germany). The stained cells were analyzed using a flow cytometer (Muse ™ Cell Analyzer, Merck Millipore, Darmstadt, Germany) and the results are shown in FIGS. 2 and 3.

As shown in FIGS. 2 and 3, when MEJG was treated at different concentrations in colorectal cancer cell line HT29 and hepatocellular cell line HepG2, the ratio of living cells decreased in a concentration-dependent manner, and the ratio of early apoptotic cells and late apoptotic cells Respectively. In other words, it was confirmed that apoptosis was increased depending on treatment concentration of MEJG. For example, in the case of HT29, early or late apoptosis occurred in 53% of the cells treated with 200 ㎍ / ㎖ of MEJG, and apoptosis was induced in approximately 60% of cells in the case of HepG2 treated with 100 ㎍ / ㎖ of MEJG there was.

Example  5: Zuernardia Grobbi Flora Trough pin  The anticancer activity-related protein of the extract Expression control ability  analysis

The expression levels of p53, p21, Bax, Bcl-2 and caspase, which are related to the apoptosis-related proteins of the extract, were analyzed by Western blot hybridization in order to clarify the mechanism of anticancer activity of MEJG prepared in Example 1 above. Specifically, the protein was extracted from the cancer cell line treated with MEJG as described above, and the protein concentration was determined using a Bradford assay. Then, 30 μg of the protein was electrophoresed on 10% SDS-PAGE, blotted on a nitrocellulose membrane, And then hybridized with the antibody to the antibody. After washing the membrane, horseradish peroxidase (HRP) was reacted with the tagged secondary antibody for 1 hour and analyzed using a chemiluminescence detection system, as shown in FIG.

As shown in FIG. 4, when MEJG was treated with cancer cell line, specifically, human colon cancer cell line HT29, the expression level of p53 as a tumor suppressor gene was not changed, but the expression of p21 as a CDK inhibitor was increased. That is, the expression of p53 was maintained at a similar level, but the expression level of p21 was increased in a p53-independent manner. In addition, the expression of Fas, a death receptor, was increased. Furthermore, the caspase cascade was activated and the expression of caspase 8 and caspase 3, active forms of caspase 8 and caspase 3, . Finally, it was confirmed that PARP (poly ADP-ribose polymerase) fragment involved in DNA repair and gene transcription, which is one of substrate proteins, was activated by the finally activated caspase 3. These results suggest that MEJG induces apoptosis of H29 by an external pathway through death receptor. In addition, the expression of Bax, a pro-apoptotic protein, and cytochrome C level in cytoplasm increased. On the other hand, it was confirmed that the expression of pro-caspase 9 was decreased due to the decrease of expression of anti-apoptotic protein Bcl-2 and the fragmentation of caspase-9. This suggests that MEJG induces apoptosis by the internal pathway through mitochondria. From these results, it was confirmed that MEJG induces apoptosis of cancer cells through two pathways of apoptosis, that is, an external pathway through death receptor and an internal pathway in mitochondria.

In summary, the extract of Zulberna diaglobrifloratrowin according to the present invention shows a killing effect specifically on cancer cells without showing toxicity to normal cells, and the Zulberna diaglobiatic florinthine extract is a cell of cancer cells Induced apoptosis, indicating that the anti - cancer effect is exhibited.

Claims (6)

A pharmaceutical composition for the prophylaxis or treatment of any cancer selected from the group consisting of lung cancer, colon cancer and liver cancer, which comprises an extract of Julbernardia globiflora as an active ingredient.
The method according to claim 1,
Wherein the extract is extracted with water, C1 to C4 lower alcohol or a mixed solvent thereof.
3. The method of claim 2,
Wherein the alcohol is methanol.
delete The method according to claim 1,
Wherein said cancer is prevented or treated by inducing apoptosis of cancer cells.
A health functional food for preventing or ameliorating any one of cancer selected from the group consisting of lung cancer, colon cancer and liver cancer, which comprises an extract of Zulberna diaglobina flora as an active ingredient.

Images