KR101862552B1 - Pharmaceutical composition, health functional food and cosmetic composition - Google Patents

Abstract

The present invention relates to a pharmaceutical composition, a health functional food, and a cosmetic composition, and more particularly, to a pharmaceutical composition for prevention or treatment of cancer, which exhibits remarkably improved anticancer effect by synergy of the combination of Tahibo extract and valproic acid, A health functional food for prevention or improvement, and a cosmetic composition for preventing or improving skin cancer.

Description

[0001] PHARMACEUTICAL COMPOSITION, HEALTH FUNCTIONAL FOOD AND COSMETIC COMPOSITION [0002]

The present invention relates to a pharmaceutical composition, a health functional food and a cosmetic composition, and more particularly, to a pharmaceutical composition for preventing or treating cancer, a health functional food for cancer prevention or improvement, and a cosmetic composition for preventing or improving skin cancer.

Cancer refers to a malignant tumor that is abnormally developed in the cell cycle of a cell that normally constitutes a human tissue and grows so that it can not regulate its growth without normally differentiating. Cancer occurs through three stages of initiation, promotion, and progression. It can occur with only a sufficient amount of carcinogens, but in practice, a small amount of initiator It is known that when a cell mutation (initiation cell) occurs due to an increase in the number of cells and an abnormal cell division by the stimulation of a tumor promoter, cancer tissue is formed.

The global cancer incidence is increasing by more than 5% annually due to the deterioration of the environment and the aging population. In 1997, the number of cancer deaths reached 6 million, or 12% of the disease death rate. In Korea, 100,000 new cases of cancer occur each year, 50,000 deaths every year, and 10% of cancer patients. According to statistics of the National Cancer Center in 1999, the number of patients was 120,000, and the order of gastric cancer (16%), liver cancer (16%), lung cancer (16%), ), Cervical cancer (12%), breast cancer (15%) and colorectal cancer (10%).

Surgical treatment, chemotherapy, biotherapy, radiation therapy, etc. have been used for the treatment of such cancers. Among them, anticancer drugs are typically used. However, most anticancer drugs currently in use are limited in use due to serious toxicity and side effects. However, there is a problem that the dosage is increased to increase the therapeutic effect, and thus it is fatal to the human body.

As such anticancer drugs, 5-fluorouracil (5-FU), doxorubicin = adriamycin, mitomycin, cisplatin and the like have been widely used. Recently, a newly developed anticancer drug Such as Paclitaxel, Docetaxel, Irinotecan, Xeloda, Oxalopatin, and the like. Among them, doxorubicin is an anticancer agent widely used for lung cancer, digestive tract cancer, etc., but it has a problem that it can cause side effects such as cardiomyopathy and the like. In addition, 5-fluorouracil (5-Fu) is a pyrimidine derivative that inhibits hexane metabolism and shows anti-cancer effects mainly on breast cancer, colon cancer, stomach cancer and pancreatic cancer, (CY) is an anti-cancer agent that inhibits DNA replication and RNA transcription and exhibits anticancer activity. It is an anticancer drug widely used for acute / chronic leukemia and other solid tumors. (Immunotoxicity), it is difficult to use it continuously. Various drugs are currently being studied to alleviate the toxicity and adverse effects of these conventional anticancer drugs.

The inventors of the present invention reached the present invention to develop an anticancer agent capable of inhibiting the development of cancer cell resistance by confirming that Tabebuia avellanedae extract has an anticancer efficacy and shows a remarkable synergistic effect when used in combination with valproic acid .

Korean Patent Publication No. 2004-0090255

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cancer, which has an improved anticancer effect according to the synergistic effect of each component.

It is an object of the present invention to provide a health functional food for cancer prevention or improvement having an improved anticancer effect.

It is an object of the present invention to provide a cosmetic composition for preventing or improving skin cancer.

1. A pharmaceutical composition for preventing or treating cancer, comprising a Tahibo extract and a valproic acid.

2. The pharmaceutical composition for prevention or treatment of cancer according to 1 above, wherein the Tahibo extract is a hot-water extract.

3. The pharmaceutical composition for preventing or treating cancer according to 1 above, wherein the Tahibo extract and the valproic acid are contained in a weight ratio of 1: 9 to 9: 1.

4. The method of claim 1 wherein the cancer is selected from the group consisting of leukemia, breast cancer, testicular cancer, bone cancer, lymphoma, metastatic melanoma, malignant melanoma, malignant glial zone, glioblastoma, brain cancer, lung cancer, colon cancer, rectum cancer, Wherein the cancer is selected from the group consisting of gastric cancer, fibrous sarcoma, smooth muscle sarcoma, rhabdomyosarcoma, liposarcoma, osteosarcoma, chondrosarcoma, angiosarcoma, lymphatic sarcoma, sebaceous sarcoma, lymphoma, A pharmaceutical composition for preventing or treating cancer.

5. The pharmaceutical composition according to above 1, wherein the cancer is leukemia, breast cancer or bone cancer.

6. A health functional food for prevention or improvement of cancer comprising Tahibo extract and valproic acid.

7. A cosmetic composition for prevention or improvement of skin cancer comprising Tahibo extract and valproic acid.

The present invention can exhibit a remarkably improved anticancer effect using a Tahibo extract and valproic acid together. Accordingly, the Tahibo extract and valproic acid can be used as a pharmaceutical composition for preventing or treating cancer that can be applied to various cancer diseases.

In addition, it can be used as a cosmetic composition for prevention or improvement of skin cancer as a health functional food for preventing or improving cancer.

FIG. 1 is a graph showing the cell survival rate of macrophages at 24 hours after treating the macrophage with the Tahibo extract of Example 1. FIG.
FIG. 2 is a graph showing experimental results showing that inhibition of cancer cell growth was observed in a concentration-dependent manner on day 4 of cultivation of Tahibo extract or valproic acid (VPA, valproic acid) in acute leukemia cell line.
FIG. 3 is a graph showing experimental results showing that inhibition of cancer cell growth was observed in a concentration-dependent manner on day 5 of the culture of tapewa extract or valproic acid (VPA, valproic acid) in acute leukemia cell lines.
FIG. 4 is a photomicrograph showing morphological changes or cell numbers of cell groups at 5 days after treatment with tapewa extract or valproic acid (VPA, valproic acid) in acute leukemia cell line.
FIG. 5 is a graph showing a comparison of growth inhibitory effects on days 4 and 5 after treatment with Tahibo extract or valproic acid (VPA, Valproic acid) in acute leukemia cell lines (A) ) Is a graph that makes it easier to see.
FIG. 6 is a graph showing a synergistic effect on cancer cell growth inhibition at 5 days after the co-treatment of Tahibo extract with valproic acid (VPA, valproic acid) in acute leukemia cell line.

The terms used in the present invention are defined as follows.

&Quot; Subject " or " patient " means any single entity for which treatment is desired, including human, cow, dog, henna pig, rabbit, chicken, It may also be included in any subject participating in a clinical trial study that does not show any disease clinical findings, or as a participant in epidemiological studies or as a control.

&Quot; Tissue or cell sample " refers to a collection of similar cells obtained from a subject or tissue of a patient. The source of the tissue or cell sample may be a solid tissue, blood or any blood component from a fresh, frozen and / or preserved organ or tissue sample or biopsy or aspirate, a cell at any time of pregnancy or development of the subject, . Tissue samples can also be primary or cultured cells or cell lines.

Optionally, tissue or cell samples are obtained from primary or metastatic tumors. Tissue samples may include compounds that are not mixed with the tissue by nature, such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, and the like. For purposes of the present invention, a " section " of a tissue sample refers to a portion or manipulation of a tissue sample, for example, a thin slice of tissue or cells sheared from a tissue sample. It will be appreciated that multiple sections of a tissue sample may be taken for analysis in accordance with the present invention if the present invention is understood to include methods in which the same section of tissue sample is analyzed both at the morphological and molecular levels, It is understood that it can do.

&Quot; Nucleic acid " is meant to include any DNA or RNA, such as chromosomes, mitochondria, viruses and / or bacterial nucleic acids present in a tissue sample. Includes one or both strands of a double-stranded nucleic acid molecule and includes any fragment or portion of the intact nucleic acid molecule.

&Quot; Gene " means any nucleic acid sequence or portion thereof that has a functional role at the time of protein coding or transcription, or in the control of other gene expression. The gene may consist of only a portion of the nucleic acid encoding or expressing any nucleic acid or protein that encodes the functional protein. The nucleic acid sequence may comprise an exon, an intron, an initiation or termination region, a promoter sequence, another regulatory sequence, or a gene abnormality within a particular sequence adjacent to the gene.

&Quot; Label " means a compound or composition that directly or indirectly facilitates the detection of a reagent, for example, a nucleic acid probe or a reagent conjugated or fused to an antibody. The label may itself be detected (e. G., A radioactive isotope label or a fluorescent label), in the case of an enzyme label, to catalyze the chemical modification of the detectable substrate compound or composition.

&Quot; Pharmaceutical composition " means a mixture of other chemical ingredients, such as a diluent or carrier, of the present invention's Tahibo extract.

&Quot; Carrier " is defined as a compound that facilitates the addition of a compound into a cell or tissue. For example, dimethylsulfoxide (DMSO) is a commonly used carrier that facilitates the introduction of many organic compounds into cells or tissues of an organism.

&Quot; Diluent " is defined as a compound that not only stabilizes the biologically active form of a compound of interest, but also dilutes it in the water in which it is dissolved. Salts dissolved in buffer solutions are used as diluents in the art. A commonly used buffer solution is phosphate buffered saline, since it mimics the salt state of the human solution. Since buffer salts can control the pH of the solution at low concentrations, buffer diluents rarely modify the biological activity of the compounds.

&Quot; Cancer ", " tumor ", or " malignant " refers to or represents the physiological condition of a mammal that is generally characterized by unregulated cell growth.

By " inhibitor " is meant a substance that inhibits, blocks or reduces the expression or activity of a particular gene. The mechanism of activation of the inhibitor is not particularly limited. Examples include organic or inorganic compounds, proteins, carbohydrates, polymeric compounds such as lipids, and composites of various compounds.

&Quot; Treatment " means an approach to obtaining beneficial or desired clinical results. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in the extent of disease, stabilization (i.e., not worsening) of the disease state, (Either partially or totally), detectable or undetected, whether or not an improvement or temporary relief or reduction Also, " treatment " may mean increasing the survival rate compared to the expected survival rate when not receiving treatment. &Quot; Treatment " refers to both therapeutic treatment and prophylactic or preventative measures. Such treatments include treatments required for disorders that have already occurred as well as disorders to be prevented. &Quot; Palliating " a disease may reduce the extent of disease states and / or undesirable clinical manifestations and / or slow the time course of progression, It means to lose.

By "about" is meant a reference quantity, level, value, number, frequency, percentage, dimension, size, quantity, weight or length of 30, 25, 20, 10, 9, 8, 7, 6, 5, 4, 3 , Level, value, number, frequency, percentage, dimension, size, quantity, weight or length of a sample,

&Quot; Pharmaceutically acceptable salt " means a formulation of a compound that does not cause severe irritation to the organism to which the compound is administered, and does not impair the biological activity and properties of the compound. The pharmaceutical salt may be prepared by dissolving the compound of the present invention in an organic solvent such as mineral acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid, sulfonic acid such as methosulfonic acid, ethanesulfonic acid, p- toluenesulfonic acid, tartaric acid, formic acid, Can be obtained by reacting with organic carboxylic acids such as acetic acid, trifluoroacetic acid, trifluoroacetic acid, capric acid, isobutanoic acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid and the like. Also, by reacting the compound of the present invention with a base, an alkali metal salt such as an ammonium salt, a sodium or potassium salt, a salt such as an alkaline earth metal salt such as a calcium salt or a magnesium salt, a dicyclohexylamine, Carmine, tris (hydroxymethyl) methylamine and the like, and amino acid salts such as arginine, lysine and the like.

&Quot; Hydrate " refers to a compound of the present invention or a salt thereof, including a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces .

&Quot; Solvate " means a compound of the present invention or a salt thereof, comprising a stoichiometric or non-stoichiometric amount of a solvent bound by noncovalent intermolecular forces. Preferred solvents therefor are volatile, non-toxic, and / or solvents suitable for administration to humans.

&Quot; Isomer " means a compound of the present invention or a salt thereof, which has the same chemical or molecular formula but is optically or sterically different. For example, the compound according to Formula 1 of the present invention may have an asymmetric center (asymmetric carbon atom) depending on the kind of substituents. In this case, the compound of Formula 1 may be an enantiomer or diastereomer May exist as the same optical isomer.

&Quot; rpodrug " means a substance that is transformed into a parent drug in vivo. Prodrugs are often used in some cases because they are easier to administer than parent drugs. For example, they may achieve viability by oral administration, whereas parent drugs may not. Prodrugs may also have improved solubility in pharmaceutical compositions over the parent drug. For example, a prodrug is an ester that facilitates the passage of a cell membrane, which is hydrolyzed to a carboxylic acid that is active by metabolism in a cell whose water solubility is once beneficial, Drug "). ≪ / RTI > Another example of a prodrug may be a short peptide (polyamino acid) that is attached to an acid group that is converted by metabolism so that the peptide reveals the active site.

Throughout this specification, the words " comprising " and " comprising ", when not explicitly required in the context, include a stated step or element, or group of steps or elements but not to any other step or element, And that they are not excluded.

Hereinafter, the present invention will be described in detail.

The pharmaceutical composition for preventing or treating cancer of the present invention includes Tahibo extract and valproic acid.

Taheebo refers to the natural tree table of the inner bark of Tabebuia avellanedae, a tree of the janissary tree native to some parts of the Amazon, or the tree itself.

The Tahibo extract may be obtained by cutting or crushing the inner bark of Tabebuia avellanedae to an appropriate size, and heating it by heating with a hot water bath. In the present invention, such a hot-water extract may be used as it is, or may be used as a purified extract after a conventional purification process including a filtration filter and chromatography. In addition, the hot-water extract and the purified extract can be used as powder by drying through processes such as vacuum distillation, freeze-drying, and natural drying.

However, the present invention is not limited to those obtained by the above method, but includes those obtained by all known processing means used for producing an ingestible form. For example, low-level ethanol extracts extracted using low-grade ethanol are also included in the Tahibo extract.

Valproic acid (VPA) has a structure represented by the following formula (1).

[Chemical Formula 1]

The pharmaceutical composition of the present invention contains the Tahibo extract and valproic acid together, so that the anticancer effect by the synergy is excellent. Therefore, even when only a small amount of each component is used, it can exhibit an excellent effect, and it is possible to suppress the intoxication and cancer cell resistance by excessive use of each component.

Valproic acid (VPA) is known to downregulate the expression of MGMT (O6-methylguanine-DNA methyltransferase). The main mechanism of MGMT gene inactivation is known as promoter methylation. This can be interpreted as the fact that valproic acid induces methylation in the target gene promoter as an HDAC inhibitor.

Many reports have shown that treatment response rates and survival rates of cancer patients with inactivated MGMT are excellent, which can be used as a material to overcome tolerance of cancer cells that are problematic in the treatment of cancer of paulinic acid It is suggesting.

Thus, inhibition of tumor cell growth of Tahibo extract can be enhanced by valproic acid in resistant cancer cells by continuous use of Tahibo. This relates to a method of using valproic acid to increase susceptibility to the anti-cancer action of Tahibo extract.

Thus, the combination of valproic acid and Tahibo extract promotes apoptosis and autophagic cell death and inhibits cell migration.

The ratio of the Tahibo extract to the valproic acid is not particularly limited, and can be used in a weight ratio of, for example, 1: 9 to 9: 1.

Unless otherwise specified, the term " Tahibo extract " or " valproic acid " as used herein refers to a concept comprising both the substance itself, its pharmaceutically acceptable salts, hydrates, solvates, isomers and prodrugs to be.

The pharmaceutical composition of the present invention may further comprise one or more extracts selected from the group consisting of mannwa (cranberry), flaxseed, and cocoa (cacao) to improve the anti-cancer effect.

The pharmaceutical composition of the present invention can arrest the growth cycle of cancer cells, inhibit growth and metastasis, or kill cancer cells.

The cancer which is a subject to be prevented or treated according to the present invention can be used for the treatment or prophylaxis of cancer such as leukemia, breast cancer, testicular cancer, bone marrow cancer, lymphoma, metastatic melanoma, malignant melanoma, malignant gliosis, glioblastoma, brain cancer, lung cancer, colon cancer, The present invention relates to a method for the treatment and prophylaxis of cancer of the esophagus, stomach cancer, fibrosarcoma, leiomyosarcoma, rhabdomyosarcoma, liposarcoma, osteosarcoma, chondrosarcoma, angiosarcoma, lymphangiosarcoma, sebaceous sarcoma, And the pharmaceutical composition of the present invention can exhibit excellent drug efficacy against cancer that has a rapid growth or metastasis or has anticancer drug resistance, preferably leukemia, breast cancer or bone marrow cancer, more preferably acute leukemia .

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, the composition of the present invention can be used in combination with or in combination with medicines such as steroidal drugs, antihistamines, antiinflammatory agents and antibiotics already used.

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.

Examples of carriers, excipients and diluents that can be contained in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium 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, Lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous agents, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As a base for suppositories, witepsol, macrogol, treen 61, cacao paper, laurin, glycerogelatin and the like can be used.

The amount of the composition of the present invention to be used may vary depending on the age, sex, and body weight of the patient, and may vary depending on the state of the subject to be treated, the specific category or type of cancer to be treated, It will depend on the sensitivity of the tumor.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

In particular, it can be administered by any medically appropriate method, for example by injection into normal cerebrospinal fluid, such as normal intravenous or intraarterial administration. In some cases, intradermal administration, intracavity administration, intrathecal administration, tumor or direct administration to the arteries supplying the tumor is advantageous. When the tumor or a portion thereof has previously been removed by surgical operation, the therapeutic agent can be delivered directly to the tumor site (and, in particular, the enclosed cavity or " ablation "Quot; resection cavity ").

Also, the present invention provides a health functional food for preventing or ameliorating cancer comprising a Tahibo extract and valproic acid.

The health functional food of the present invention may further comprise the above-mentioned Tahibo extract and a pharmaceutically acceptable food-aid additive, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed is, for example, , Prevention, health, or therapeutic treatment.

The effective amount of Tahibo extract and valproic acid contained in the health functional food may be used in accordance with the effective amount of the pharmaceutical composition. However, in the case of long-term intake for the purpose of health and hygiene or health control, And the active ingredient has no problem in terms of stability, so that it can be used in an amount exceeding the above range.

There is no particular limitation on the kind of the health functional food. Examples of the health functional food include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen and other noodles, gums, ice cream, , Beverages, tea, a drink, an alcoholic beverage and a vitamin complex, and a health functional food.

The present invention also provides a cosmetic composition for prevention or improvement of skin cancer comprising a Tahibo extract and valproic acid.

The cosmetic composition of the present invention may further contain ingredients commonly used in cosmetic compositions in the art besides Tahibo extract and valproic acid. For example, it may further contain fragrance, dye, antioxidant, moisturizer, thickening agent, excipient, diluent, inorganic salt and synthetic polymer material, and the content may be appropriately adjusted .

The cosmetic composition according to the present invention can be formulated into an external preparation for skin or cosmetics. The external preparation for skin may be an ointment, a warning agent, a spray, a suspension, an emulsion, a cream, a gel or the like. The cosmetics may include basic cosmetics, makeup cosmetics, body cosmetics, shaving cosmetics, and the like. Examples of the basic cosmetics include a cream, a lotion, a pack, a massage cream, and an emulsion. Examples of the makeup cosmetics include a foundation, a makeup base, a lipstick, an eye shadow, an eyeliner, a mascara, an eyebrow pencil, Examples of cosmetics include soaps, liquid cleansers, bath salts, sunscreen creams, sun oils, and examples of shaving cosmetics include aftershave lotions and shaving creams.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to examples.

Example  One. Tahibo  Preparation of extract

1 kg of the inner bark of Tabebuia avellanedae was cut or pulverized to an appropriate size and extracted by heating in a hot water bath at 85 to 95 ° C for 2 to 3 hours. The obtained hot-water extract was purified using a filter. The purified extract was dried and concentrated under reduced pressure using a freeze dryer to obtain a Tahibo extract. The dried Tahibo extract was stored at room temperature.

Example  2. Cell culture

The cell line used to confirm the safety of the normal cell was RAW264.7 macrophage. The cell line used to identify the anticancer effect was a cell line that induced acute myelogenous leukemia obtained by inserting and expressing the MN1 gene in the cell to be. (100 mm dish) using a culture medium RPMI1640 (Gibco), 10% FBS (fetal bovine serum, HyClone) and an antibiotic (1/100 penicillinstreptomycin, Gibco) CO ₂ incubator.

Example  3. Cell survival rate of normal cells

The Tahibo extract obtained in Example 1 was prepared at each concentration (0, 50, 100, 200, 400 ㎍ / ml) and then cultured in macrophage RQW264.7 (1x10 ⁶ cells / ml) Respectively. And cultured for 24 hours after the treatment. 10 μl of MTT solution (10 mg / ml in phosphate buffered saline (PBS), pH 7.4) was added to the cultured cells and after 3 hours, 15% SDS (Sodium dodecylsulfate) was added to terminate the reaction. The cytotoxicity test of the Tahibo extract was performed by a conventional MTT assay (Lee, JY et al., 2007. Hydroquinone, a reactive metabolic of benzene, and reduced macrophage-mediated immune responses. Mol Cells 23: 198-206) The optical density at 570 nm, i.e. absorbance, was measured using a SpectraMax 250 microplate reader (FIG. 1).

As shown in Fig. 1, the Tahibo extract showed almost no cytotoxicity even at a concentration of 400 占 퐂 / ml. However, it showed slight cytotoxic effect at 400 ㎍ / ml concentration, but there was no statistically significant difference.

Example  4. In acute leukemia cells Tahibo  Extract or VPA Growth inhibition effect

Cells at a density of 5x10 ³ cells / well in 12-well plate being said, 15% FBS and stem cell factor (20 ng / ml ), IL-6 (10 ng / ml), IL-3 (6 ng / ml ) In DMEM medium for 4 hours. Then, the Tahibo extract was treated at a concentration of 0, 100, 400 占 퐂 / ml, or in a concentration of 100 占 퐂 / ml (0.7 mM) and 250 占 퐂 / ml (1.7 mM) . After the treatment, the number of living cells was measured on the fourth or fifth day of culture.

(1) Effects of singly

First, in order to investigate the effect of Tahibo extract or valproic acid alone in acute leukemia cell line, 100 μg or 400 μg of Tahibo extract was treated with 100 μg or 400 μg on the basis of 1 ml medium, or 100 μg or 250 μg of valproic acid was treated. 100 and 250 μg / ml of valproic acid correspond to concentrations of 0.7 and 1.7 mM, respectively.

As shown in FIG. 2, the number of living cells on day 4 was measured. As a result, about 40% of tumor cells were inhibited by treatment with 100 μg of Tahibo extract. In the case of treatment with 400 μg of Tahibo extract, , Respectively. On the other hand, in the group treated with 100 μg (0.7 mM) of valproic acid, the growth inhibitory effect was about 9%, but there was no statistically significant difference. In the group treated with 250 μg (1.7 mM) Growth inhibitory effect.

Respectively, showed inhibitory effects on cancer cell growth, even when treated with Tahibo extract or valproic acid alone.

As shown in FIG. 3, in the treatment group of 100 μg of Tahibo extract, about 56% of the cancer cell growth inhibitory effect was observed. In the treatment group of 400 μg of Tahibo extract, about 97% of the growth Inhibitory effect. On the other hand, in the group treated with 250 ㎍ (1.7 mM) of valproic acid, the growth inhibitory effect was about 80%.

As shown in FIG. 2 and FIG. 3, it was shown that each treatment of tumor extract or valproic acid alone inhibited cancer cell growth.

Morphological observation was also carried out using a microscope (Fig. 4). Microscopic observations showed that the number of cells in the treatment group of 400 μg of Tahibo extract was significantly decreased, and most of the cancer cells showed atypical morphology and many dead cells.

FIG. 5 shows changes over time, comparing the 4th day and 5th day of culturing (FIG. 5A, B). It was found that cancer cells did not proliferate even if they were cultured for one more day in the Tahoe extract treatment group or the valproic acid treatment group. On the other hand, the number of cells in the treatment group of 400 μg of Tahibo extract and 250 μg of valproic acid (1.7 mM) decreased on average. This means showing the ability of the Tahibo extract or valproic acid to inhibit the cell-cycle arrest of cancer cells.

(2) Effect of combination

Next, in order to evaluate the synergy effect of using Tahibo extract and valproic acid in acute leukemia cells, 100 μg of Tahibo extract, 100 μg of valproic acid, 100 μg of Tahibo extract, And 100 ㎍ of furonic acid were compared.

In the case of the single effect, 5,000 cells were added to the plate, and in order to confirm whether the same effect was obtained even when the number of cells was increased, the experiment was conducted under the condition that 20,000 cells were added to the first number of cells added in the combination experiment .

As shown in FIG. 6, the inhibition of cancer cell growth by about 40% was observed when treating the Tahibo extract alone, but there was no statistical significance when treated with valproic acid (VPA) alone .

On the other hand, the group treated with 100 μg of Tahibo extract and 100 μg of valproic acid showed a much higher synergistic effect (99% inhibition rate) than the cell groups treated alone, And the inhibitory effect of the two substances is shown to be much higher than the synergistic effect (synergy effect) is shown.

Claims (7)

A pharmaceutical composition for prevention or treatment of leukemia, comprising a Tahibo extract and valproic acid as an active ingredient.
[Claim 2] The pharmaceutical composition for preventing or treating leukemia according to claim 1, wherein the Tahibo extract is a hot-water extract.
The pharmaceutical composition for preventing or treating leukemia according to claim 1, wherein the Tahibo extract and valproic acid are contained in a weight ratio of 1: 9 to 9: 1.
delete delete Takiho extract and valproic acid as an active ingredient, a health functional food for preventing or improving leukemia. delete

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