KR101465238B1 - Composition for preventing or treating of cancer comprising 4-isopropyl-2,6-bis(1-phenylethyl)phenol from Cordyceps bassiana as an active ingredient - Google Patents

Abstract

The present invention relates to a composition comprising 4-isopropyl-2,6-bis(1-phenylethyl)phenol derived from Cordyceps bassiana as an active ingredient for preventing or treating cancer and, more specifically, to 4-isopropyl-2,6-bis(1-phenylethyl)phenol which concentration-dependently inhibits proliferation of cancer cells in RAW264.7 cells, C6 cells, MDA-MB-231 cells, and A549 cells; induces apoptosis by activating caspase-8 and -7; and effectively suppresses the formation of tumors and increases the survival rate in a tumor transplantation mice model. Thus, the 4-isopropyl-2,6-bis(1-phenylethyl)phenol of the present invention can be usefully applied as an active ingredient for a composition or a functional health food for preventing and treating cancer.

Description

The present invention relates to a composition for prevention or treatment of cancer comprising 4-isopropyl-2,6-bis (1-phenylethyl) phenol as an active ingredient, 6-bis (1-phenylethyl) phenol from Cordyceps bassiana as an active ingredient}

The present invention includes 4-isopropyl-2,6-bis (1-phenylethyl) phenol as an active ingredient, which is a component derived from yellow-rooted caterpillar fungus Or a pharmaceutical composition for preventing or treating cancer, or a health food containing the same.

Despite the development of modern science and technology, along with the development of medical technology, biotechnology and genetic engineering, various researches and methods for treating cancer have been developed, but cancer is still recognized as a serious disease that can not be cured. Many methods such as incision, chemotherapy, radiotherapy, and incision have been developed to treat these cancers, but radiation therapy or incision is mainly effective only when the initial diagnosis of cancer is made, It is of no use and can only be treated with chemotherapy. Since the 1940s, the chemotherapy introduced into cancer therapy has been attracting much attention because of its advantage that it can be applied relatively easily regardless of the timing of cancer, and various chemotherapeutic agents have been developed.

However, when the above-mentioned anticancer drugs are administered repeatedly for a long period or when the cancer is recurred, the cancer cells lose the therapeutic effect by acquiring resistance to the anticancer drug. In addition, most anticancer drugs have the effect of inhibiting the synthesis of nucleic acid in the cell or directly binding to nucleic acid to impair its function. These anticancer drugs do not only act selectively on cancer cells, but also on normal cells, Which causes various side effects such as a decrease in bone marrow function, gastrointestinal mucosal damage, hair loss, and the like.

Recently, alternative medical treatments such as Chinese traditional medicine have been rapidly increasing not only in the western countries but also in Asia. So far, many natural compounds derived from nature have been shown to exhibit anticancer effects induced by apoptosis, and these natural compounds can be used as new and effective natural compounds based on the discovery of new drugs for the treatment and prevention of cancer (Athar et al., 2007; Nobili et al., 2009).

Apoptosis is known to be a programmed cell death, a genetic control mechanism that plays an important role in maintaining cell homeostasis. Apoptosis can generally be induced through an exogenous (death receptor) pathway or endogenous (in mitochondria) pathway. In the exogenous pathway, the ligation of the TNF / Fas-receptor by its ligand induces cleavage of the caspase-8 initiator and directly activates the effector caspase-3 or activates Bcl-2 (Mellier et al., 2010; Tan et al., 2009; Wu, 2009) induces the disruption of Bax in the mitochondrial membrane after inducing cleavage of the family member Bid. On the other hand, the endogenous pathway is mediated by mitochondria, and in response to aortotoxic stimuli, cytochrome c and apoptosis-inducing factor (AIF) are released from the mitochondria and these factors are caspase-dependent and independent It is involved in apoptosis. Cytochrome c binds to Apaf-1 to form a structure called apoptosome, activates caspase-9, then activates caspase-3, and eventually causes apoptotic cell death . AIF is a marker of caspase-independent apoptosis, which, after apoptotic stimulation, translocates into the nucleus and induces DNA fragmentation (Huerta et al, 2006; Millan and Huerta, 2009).

On the other hand, it is a generic term for all the fungi which are parasitic on insects, arthropods, fungi or seeds of higher plants. Mycologically, it belongs to the acanthaceous germs, incomplete germs (incomplete germs), and junctional germs (germs). Up to now, about 800 fungi have been reported, and 76 fungi have been isolated and identified in Korea.

Cordyceps has been used as an antidote to treatment of tuberculosis and asthma jaundice and opioid poisoning, as a remedy for obesity, and as a supplemental tonic and immunity enhancer. In addition, it has been reported that the herbal product is effective in the treatment of kidney and pulmonary diseases, and it has the effect of strengthening the fragile constitution and increasing the immunity. In addition, the Chinese Taipei Taechangwacho is sweet and mild and makes the lung strong, so it is said that it is good for respiratory diseases. In recent years, there have been a lot of studies on the development of therapeutic agents and medicines using cocoon extracts in Korea. However, it is very difficult to collect caterpillar fungus in its natural state. Even if it is collected, very small amount of fungicide is collected. Therefore, it is difficult to develop a product using only collected organisms and to search for pharmacological components and new substances beneficial to human body.

Cordyceps, one of the species of Cordyceps, bassiana ) is a type of Entomopathogenic Fungi that enters the insect and forms a fruiting body by its host. The taxonomic positions are ascomycota, Pyrenomycetes, Clavicipitales, Cordycipitaceae, , Cordyceps (Cordyceps) belong to the incomplete generation. It was first reported in China and was found in Korea. Beauveria , a form that appears in the life cycle of caterpillar fungus, bassiana has long been used in the treatment of neurological diseases, cancer, skin infections, wounds and the like in China and Korea (Donguibogam). However, 4-isopropyl-2,6-bis (1- 4-isopropyl-2,6-bis (1-phenylethyl) phenol) has not been known at all for the human body.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a process for producing 4-isopropyl-2,6-bis (1-phenylethyl) 6-bis (1-phenylethyl) phenol) inhibits the proliferation of cancer cells and inhibits tumor growth. Thus, it has been shown that 4-isopropyl-2,6- And the like.

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

In order to achieve the above object, the present invention provides a process for preparing 4-isopropyl-2,6-bis (1-phenylethyl) phenol, Or a pharmaceutically acceptable salt thereof as an active ingredient, or a health food containing the same.

In one embodiment of the present invention, the 4-isopropyl-2,6-bis (1-phenylethyl) phenol is obtained from Cordyceps bassiana ).

In another embodiment of the present invention, the cancer is characterized in that it is selected from the group consisting of glioma glioma, breast cancer and lung cancer.

In another embodiment of the present invention, the 4-isopropyl-2,6-bis (1-phenylethyl) phenol increases the activity of Caspase-8 or Caspase-7 protein in cancer cells to induce apoptosis .

The 4-isopropyl-2,6-bis (1-phenylethyl) phenol of the present invention inhibits the proliferation of cancer cells in various cancer cell lines, increases the activity of signal transduction proteins that induce apoptosis of cancer cells However, it can exhibit an inhibitory effect on tumor formation even in vivo, and thus can be usefully used for prevention or treatment of cancer.

As far as the effects on 4-isopropyl-2,6-bis (1-phenylethyl) phenol have not been studied so far, 4-isopropyl- Can be used not only for the purpose of attempting a new scientific study on cancer diseases but also for the development of medicines and quasi drugs.

FIG. 1 is a schematic diagram showing a process for obtaining an extract or a fraction using a yellow-leafy Cordyceps;
FIG. 2 is a schematic view showing a process of performing secondary silica chromatography using a butanol fraction of a yellow copal cordworm.
FIG. 3 shows that among the 18 peaks obtained from the secondary chromatography fractions of the butanol fraction of the yellow copalia cordata extract, the Si2-100-4 fraction, the Si2-90-1 fraction, and the Si2-90-2 fraction were combined, (1-phenylethyl) phenol (KTH-13) was confirmed in the second peak.
4 shows the results of ¹ H-NMR data of 4-isopropyl-2,6-bis (1-phenylethyl) phenol of the present invention.
FIG. 5 shows the results of confirming the inhibitory effect of 4-isopropyl-2,6-bis (1-phenylethyl) phenol on cancer cell proliferation in various cancer cell lines.
6 is a result of confirming the activity of inducing the activity of caspase-8 protein of 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13).
FIG. 7 shows the activity-inducing effect of caspase-7 protein of 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13).
FIG. 8 shows the results of confirming the inhibitory effect of 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) on tumorigenesis in tumor transplanted mice.
FIG. 9 shows the results of confirming the effect of 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) on the survival rate of tumor-transplanted mice.

The present inventors have found that the relationship between 4-isopropyl-2,6-bis (1-phenylethyl) phenol and 4-isopropyl-2,6-bis (1-phenylethyl) phenol, which is a component derived from yellow- We could do infinite proliferation as RAW264.7 cells (macrophages), C6 cells (glioma cancer cells), MDA-MB-231 cells (breast cancer cells) and A549 cells (lung cancer cells) When the cell line that differentiates is treated with 4-isopropyl-2,6-bis (1-phenylethyl) phenol, the proliferation of cancer cells is inhibited in a concentration-dependent manner, and caspase-8 and- 4-isopropyl-2,6-bis (1-phenylethyl) phenol was found to inhibit tumor growth by confirming that it not only induces apoptosis but also effectively inhibits tumor growth and increases its survival rate in tumor- The present invention can be effectively used as an active ingredient of a composition for prevention and treatment or a health food. Completed.

Hereinafter, the present invention will be described in detail.

The present invention relates to a pharmaceutical composition containing 4-isopropyl-2,6-bis (1-phenylethyl) phenol or a pharmaceutically acceptable salt thereof as an active ingredient Or a pharmaceutically acceptable salt thereof.

The 4-isopropyl-2,6-bis (1-phenylethyl) phenol can be used as a natural product or a compound produced by synthesis through ordinary methods of synthesis and fractionation of substituents, ( Cordyceps bassiana , but is not limited thereto.

The 4-isopropyl-2,6-bis (1-phenylethyl) phenol of the present invention can be used in the form of a pharmaceutically acceptable salt. The salt may be formed by a pharmaceutically acceptable free acid Acidophosphate is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Dioleate, aromatic acid, aliphatic and aromatic sulfonic acids. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene sulfide Propyl sulphonate, naphthalene-1-yne, xylenesulfonate, phenylsulfate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, Sulfonate, naphthalene-2-sulfonate or mandelate.

The acid addition salt according to the present invention can be prepared by dissolving 4-isopropyl-2,6-bis (1-phenylethyl) phenol in an excess amount of an acid aqueous solution, For example, methanol, ethanol, acetone or acetonitrile.

By heating the same amount of 4-isopropyl-2,6-bis (1-phenylethyl) phenol and an acid or alcohol in water, then evaporating the mixture and drying, or by suction filtration of the precipitated salt .

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. In addition, the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

In addition, the 4-isopropyl-2,6-bis (1-phenylethyl) phenol of the present invention can be used not only for pharmaceutically acceptable salts, but also for all salts, hydrates and solvates .

The addition salt according to the present invention can be prepared by a conventional method. For example, 4-isopropyl-2,6-bis (1-phenylethyl) phenol is dissolved in a water-miscible organic solvent such as acetone, methanol, ethanol , Or acetonitrile, etc., adding an excess amount of an organic acid, or adding an aqueous acid solution of an inorganic acid, followed by precipitation or crystallization. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

The cancer may be selected from the group consisting of liver cancer, stomach cancer, breast cancer, colon cancer, bone cancer, pancreatic cancer, head or neck cancer, uterine cancer, colon cancer, lung cancer, ovarian cancer, rectum cancer, esophageal cancer, small intestine cancer, fallopian tube carcinoma, endometrial carcinoma, The present invention is preferably any one selected from the group consisting of glioma, vulvar carcinoma, vulvar carcinoma, Hodgkin's disease, prostate cancer, bladder cancer, kidney cancer, , But is not limited thereto.

Preferably, the 4-isopropyl-2,6-bis (1-phenylethyl) phenol induces apoptosis by increasing the activity of Caspase-8 or Caspase-7 protein in cancer cells, but is not limited thereto.

In a specific example of the present invention, when 4-isopropyl-2,6-bis (1-phenylethyl) phenol is treated with RAW264.7 cells, C6 cells, MDA-MB-231 cells and A549 cell lines, Inhibit the proliferation of cancer cells in dependence on the cancer cell concentration, activate caspases -8 and -7 to induce apoptosis, and effectively inhibit tumor growth in tumor transplantation model mice, Respectively.

Accordingly, the 4-isopropyl-2,6-bis (1-phenylethyl) phenol of the present invention effectively induces apoptosis of cancer cells in various cancer cell lines, and thus can be effectively used as an effective ingredient of a composition for preventing or treating cancer have.

When the composition of the present invention is used as a pharmaceutical composition, a pharmaceutical composition containing 4-isopropyl-2,6-bis (1-phenylethyl) phenol or a pharmaceutically acceptable salt thereof as an active ingredient, May be formulated and administered in a variety of oral or parenteral dosage forms such as, but not limited to,

Examples of the formulations for oral administration include tablets, pills, light / soft capsules, liquids, suspensions, emulsions, syrups, granules and elixirs. These formulations may contain, in addition to the active ingredient, a diluent (e.g., lactose, dextrose, (E.g., silica, talc, stearic acid and magnesium or calcium salts thereof and / or polyethylene glycols), such as, for example, water, rosin, sucrose, mannitol, sorbitol, cellulose and / or glycine. The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine and may optionally contain additives such as starch, agar, alginic acid or its sodium salt A disintegrating or boiling mixture and / or an absorbent, a colorant, a flavoring agent, and a sweetening agent.

The pharmaceutical composition containing 4-isopropyl-2,6-bis (1-phenylethyl) phenol as an active ingredient can be administered parenterally, and parenteral administration can be carried out by subcutaneous injection, intravenous injection, intramuscular injection, It depends on the injection method.

At this time, 4-isopropyl-2,6-bis (1-phenylethyl) phenol or a pharmaceutically acceptable salt thereof is mixed with water together with a stabilizer or a buffer to formulate a formulation for parenteral administration, And can be prepared in an ampoule or vial unit dosage form. The composition may be sterilized or may contain other therapeutically useful substances such as preservatives, stabilizers, wettable or emulsifying accelerators, salts for controlling osmotic pressure and / or buffers, and other therapeutically useful substances, It can be formulated according to the coating method.

The dose of the compound of the present invention to the human body may vary depending on the patient's age, weight, sex, dosage form, health condition, and disease severity. In general, when referring to an adult patient weighing 60 kg, And may be 0.01 to 500 mg / day, preferably 0.01 to 500 mg / day, and may be administered once or several times a day at regular intervals according to the judgment of a doctor or pharmacist.

The present invention also relates to the use of 4-isopropyl-2,6-bis (1-phenylethyl) phenol or a pharmaceutically acceptable salt thereof as an active ingredient Or a pharmaceutically acceptable salt thereof.

The 4-isopropyl-2,6-bis (1-phenylethyl) phenol can be used as a natural product or a compound produced by synthesis through ordinary methods of synthesis and fractionation of substituents, ( Cordyceps bassiana , but is not limited thereto.

The cancer may be selected from the group consisting of liver cancer, stomach cancer, breast cancer, colon cancer, bone cancer, pancreatic cancer, head or neck cancer, uterine cancer, colon cancer, lung cancer, ovarian cancer, rectum cancer, esophageal cancer, small intestine cancer, fallopian tube carcinoma, endometrial carcinoma, The present invention is preferably any one selected from the group consisting of glioma, vulvar carcinoma, vulvar carcinoma, Hodgkin's disease, prostate cancer, bladder cancer, kidney cancer, , But is not limited thereto.

The 4-isopropyl-2,6-bis (1-phenylethyl) phenol of the present invention inhibits the proliferation of cancer cells and effectively induces apoptosis in various cancer cell lines, Bis (1-phenylethyl) phenol can be usefully used as an active ingredient of a health food for cancer prevention or improvement.

The health food of the present invention may contain various flavors or natural carbohydrates as an additional ingredient. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The natural carbohydrate is preferably selected from the range of 0.01 to 0.04 part by weight, preferably about 0.02 to 0.03 part by weight per 100 parts by weight of the health food of the present invention, but is not limited thereto.

In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. These components may be used independently or in combination. The proportion of such additives is not critical, but is preferably selected from the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health food of the present invention, but is not limited thereto.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

[ Example ]

Example  1. From the yellow mulberry caterpillar fungus, 4-isopropyl-2,6- Bis (One- Phenylethyl )phenol( KTH -13)

In order to extract the effective substances of yellow mulberry caterpillars from brown rice, 360 g of yellow mulberry caterpillar fungus was extracted with ethanol to obtain 66.1 g (19% yield) of ethanol extract. Then, 66.1 g of the ethanol extract was fractionated successively with water, hexane, butanol and ethyl acetate, and the specific steps of obtaining each fraction are shown in the schematic diagram of FIG.

As a result, 63.4% of water fraction, 22% of hexane fraction, 7.3% of butanol fraction and 7.3% of ethyl acetate fraction were obtained. The butanol fraction of the obtained fractions showed the best result. The fractions were separated and purified by secondary silica chromatography.

The fractions (Si2-100-4 fraction, Si2-90-1 fraction and Si2-90-2 fraction) showing the result of high physiological activity (anti-cancer activity) among the separated fractions were combined and finishing was carried out. Among the 18 peaks, 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) was confirmed at the 11th peak (FIG.

Example  2. Production of 4-isopropyl-2,6-bis (1- Phenylethyl )phenol( KTH -13)

Cordyceps 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13), which is a component derived from bassiana , Respectively.

[constitutional formula]

Specifically, 4-isopropylphenol (68 mg, 0.5 mmol), 1-phenylethanol (134.4 mg, 1.1 mmol) and p-toluenesulfonic acid (3 mmol) were added in a Shinkle flask equipped with a magnetic stir bar Drop) was dissolved in anhydrous CH ₂ Cl ₂ (5 mL) under a nitrogen atmosphere. The mixture was refluxed overnight. The mixture was cooled at room temperature and extracted with CH ₂ Cl ₂ / H ₂ O (saturated NaHCO ₃ ). Separating the organic layer was dried over anhydrous MgSO _4. The solution was filtered and evaporated. The crude compound was then purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane, 9: 1). Yield: 53 mg (31%). The compound is present as a partial isomer because of the chiral center of the phenylethyl group.

(S, 1H), 4.30 (q, J = 7.2 Hz, 2H), 2.87 (s, 2H) J = 6.9 Hz, 1H), 1.59 (d, J = 7.2 Hz, 6H), 1.26 (d, J = 6.9 Hz, 6H).

(S, 1H), 4.28 (q, J = 7.2 Hz, 2H), 2.87 (s, 2H) J = 6.9 Hz, 1H), 1.58 (d, J = 7.2 Hz, 6H), 1.23 (d, J = 6.9 Hz, 6H).

Example  3. 4-Isopropyl-2,6- Bis (One- Phenylethyl ) Phenol (KTH-13) inhibited cancer cell proliferation

In order to examine the effect of 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) of the present invention on cancer cell proliferation in various cancer cell lines, KTH-13 Next, cell viability of each cancer cell line was confirmed.

Specifically, RAW264.7 cells, C6 giloma cells, MDA-MB-231 cells (breast cancer cells), A549 cells (lung cancer cells) (the American Type Culture Collection (Manassas, Va.)) Were plated at 1 × 10 ⁵ , respectively, and KTH-13 was treated by concentration. At this time, KTH-13 was treated with 0 to 400 μM of RAW 264.7 cells, and with 0 to 100 μM of CEDCs, MAD-MB-231 cells and A549 cells. After incubation at 37 ° C in a 5% incubator for 48 hours, MTT assay was performed. The MTT assay was performed by incubating the MTT solution with MTT solution for 3 hours and stop solution at 37 ° C in a 5% incubator for 18 hours and then measuring the optical density at 405 nm.

As a result, as shown in FIG. 5, when KTH-13 was treated, it was confirmed that the proliferation of each cancer cell line was inhibited in a KTH-13 concentration-dependent manner.

Example  4. 4-Isopropyl-2,6- Bis (One- Phenylethyl ) Phenol (KTH-13) Caspase ( Caspase ) Identification of active induction

Since caspase plays an essential role in apoptosis, the proliferation inhibitory effect of 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) In order to confirm whether or not it is related to the activity of caspase, C6 cells were treated with 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) and the activity of caspase was measured .

Specifically, caspase-8, which regulates apoptosis, was treated with 50 μM KTH-13 to C6 cells, and after 12 hours, a protein sample was isolated from the cells, and polyclonal anti-caspase-8 antibody (Cell Signaling, Beverly Immunoblotting was performed by the method of Byeon et al. (2013, Free Radical Biology and Medicine, Apr.

In the case of caspase-7 and caspase-3, C6 cells were treated with 50, 75 and 100 μM of KTH-13, and after 6 hours, the membrane was dissolved in the resulting cells to separate protein samples. Polyclonal anti-caspase-7 and anti Immunoblotting was performed by the method of Byeon et al. (2013, Free Radical Biology and Medicine, Apr. 57: 105-18) using an anti-caspase-3 antibody (Cell Signaling, Beverly, .

6 and 7, the degree of activation of caspase-8 and caspase-7 by the treatment with 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) Cleaved Caspase-8 and Cleaved Caspase-7, which are active proteins, were increased.

Thus, the present 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) inhibits cancer cell proliferation by increasing the activity of a signal transduction protein And it has an effect of promoting the death.

Example  5. 4-Isopropyl-2,6- Bis (One- Phenylethyl ) Phenol (KTH-13) inhibitory effect on tumor formation in mouse

In order to confirm whether the 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) of the present invention shows tumor suppression effect in vivo, tumor- -2,6-bis (1-phenylethyl) phenol (KTH-13).

Specifically, a xenograft cancer model in which RMA cells (1 × 10 ⁶ cells / mouse) were injected into C57BL / 6 mouse (Daehan Biolink, Chungbuk, Korea) was used. First, 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) was dissolved in 0.5% CMC (Carboxymethyl Cellulose) before the Xenograft cancer model was constructed. vehicle treated group) and the experimental group (compound treated group), and then orally twice a day for 7 days. After 7 days of oral administration, RMA cells were subcutaneously injected into each group of mice to construct a Xenograft cancer model. The size of the cancer tissues was determined for 30 days. Then, mouse survival rate after 45 days was confirmed and compared.

As a result, as shown in FIGS. 8 and 9, when the mouse treated with 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) The development of tumors was suppressed, the rate of tumor generation was remarkably decreased, and the survival rate was remarkably increased.

Thus, it can be seen that the 4-isopropyl-2,6-bis (1-phenylethyl) phenol (KTH-13) of the present invention shows an excellent tumorigenic inhibitory effect in vivo.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

Claims (8)

Cancer prevention comprising 4-isopropyl-2,6-bis (1-phenylethyl) phenol or a pharmaceutically acceptable salt thereof as an active ingredient Or a pharmaceutically acceptable salt thereof.
The method of claim 1, wherein the 4-isopropyl-2,6-bis (1-phenylethyl) phenol is selected from the group consisting of Cordyceps bassiana . < / RTI >
The composition of claim 1, wherein the cancer is selected from the group consisting of glioma, breast, and lung cancer.
The method according to claim 1, wherein the 4-isopropyl-2,6-bis (1-phenylethyl) phenol increases the activity of Caspase-8 or Caspase-7 protein in cancer cells to induce apoptosis , Composition.
Cancer prevention comprising 4-isopropyl-2,6-bis (1-phenylethyl) phenol or a pharmaceutically acceptable salt thereof as an active ingredient Or improving health food.
The method of claim 5, wherein the 4-isopropyl-2,6-bis (1-phenylethyl) phenol is selected from the group consisting of Cordyceps bassiana ). < / RTI >
6. The health food according to claim 5, wherein the cancer is selected from the group consisting of glioma, breast cancer and lung cancer.
6. The method according to claim 5, wherein the 4-isopropyl-2,6-bis (1-phenylethyl) phenol increases the activity of Caspase-8 or Caspase-7 protein in cancer cells to induce apoptosis , Health food.

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