KR101803046B1 - Anti-cancer composition comprising alcohol extracts of Selaginella tamariscina as an active ingredient for combinational administration with chemotherapeutics - Google Patents

Abstract

In this specification, an anticancer composition comprising an organic solvent extract of Buchucson as an active ingredient is disclosed. In one aspect, a composition containing bouchalcohol alcohol extract as an active ingredient can enhance the anticancer effect of a chemotherapeutic agent when it is administered in combination with a chemotherapeutic agent. When the composition is administered in combination, an excellent anti-cancer effect can be obtained even when a small amount of an anticancer agent is administered. Therefore, it is possible to reduce the dose of cytotoxic anticancer drug to healthy cells in addition to cancer cells, thereby effectively treating cancer while maintaining the health of cancer patients.

Description

[0001] The present invention relates to an anticancer composition for use in combination with a chemotherapeutic agent comprising an extract of Buchoes Alcohol as an active ingredient,

In the present specification, a composition for promoting anticancer efficacy comprising brucus alcohol extract as an active ingredient is disclosed.

Although cancer research has been conducted for a long period of time, there are still more than 10 million cancer cases worldwide every year, and cancer incidence is continuously increasing due to environmental pollution and erroneous eating habits. Cancer patients receive chemotherapy or receive surgical treatment through cancer surgery.

Cancer chemotherapy not only affects cancer cells but also normal tissues (bone marrow, gastrointestinal mucosa, reproductive system, hair, etc.) which are active in cleavage, and sometimes cause secondary cancer. Typical side effects include anemia, decreased white blood cell / platelet count, stomatitis, nausea / vomiting, diarrhea, and hair loss. In the case of continuous administration of anticancer drugs, cancer cells express various drug resistance proteins which avoid the toxicity of anticancer drugs, thereby acquiring drug resistance to anticancer drugs, which makes chemotherapy difficult. Therefore, there is a need for a method of lowering the prescription dose of an anticancer drug used for chemotherapy, and a method of administering a cancer patient safely after cancer therapy, while maintaining the anticancer effect. Recently, there is a strong social need for anticancer therapy using natural products. The Ministry of Health and Welfare, the National Cancer Center, and the National Cancer Information Center are publishing data on cancer therapy supplementation and alternative therapies, and anticancer supplements such as Sanjia, which have anticancer effects in the private sector, continue to be introduced. However, it is still limited to indiscriminate use of antitumor supplements due to hepatotoxicity and induction of renal toxicity. Therefore, it is essential to develop a natural-derived anticancer adjuvant that has proved scientific evidence of the cancer treatment effect harmless to the human body.

Ting-Chao Chou, Pharmacological Reviews, 58, 621-681, 2006

In one aspect, an object of the present invention is to provide a natural cancer-derived anticancer composition which can be co-administered with a chemotherapeutic agent.

In another aspect, an object of the present invention is to obtain an excellent anti-cancer effect while reducing the amount of a chemotherapeutic agent administered to a human body.

In another aspect, an object of the present invention is to provide a cancer treatment composition in which the anticancer effect is enhanced.

In one aspect, the present invention provides an anticancer composition comprising an organic solvent extract of Buchucson as an active ingredient.

In one aspect, the present invention provides an anticancer composition comprising, as an active ingredient, a chemotherapeutic agent and an organic solvent extract of Bukhandan.

In another aspect, the present invention provides a composition comprising an organic solvent extract of Buchucson, a composition comprising a chemotherapeutic agent, and an instruction sheet, wherein the Buchucson organic solvent extract and the chemotherapeutic agent are mixed at a weight ratio of 1: As an anti-cancer agent.

In one aspect, a composition containing an extract of bushsohn alcohol as an active ingredient can improve the anticancer effect of a chemotherapeutic agent when it is administered in combination with a cancer treatment chemotherapeutic agent. When the composition is administered in combination, an excellent anti-cancer effect can be obtained even when a small amount of an anticancer agent is administered. Therefore, it is possible to reduce the dose of cytotoxic anticancer drug to healthy cells in addition to cancer cells, thereby effectively treating cancer while maintaining the health of cancer patients.

FIG. 1A is a graph showing the inhibitory effect of the human lung cancer cell line A549 on the growth of human lung cancer cell line according to the concentration of the administered substance, when ethanol extract of Buchoeson alone, doxorubicin alone, and Buchoeson ethanol extract and doxorubicin were administered.
FIG. 1B shows a combination index value obtained by confirming the synergistic action of Buchoes' ethanol extract and doxorubicin.
FIG. 2 is a graph showing the inhibitory effect of human lung cancer cell line A549 on the growth of human lung cancer cell line according to the concentration of the administered substance when administered alone or in combination with doxorubicin and doxorubicin combined with Buchoeson water extract.
FIG. 3 is a graph showing the inhibitory effect of NCI-H460 on the growth of the human lung cancer cell line according to the concentration of the administered substance, when the Buchoeson ethanol extract alone, the doxorubicin alone, and the Buchoeson ethanol extract and the doxorubicin combination were administered.
FIG. 4 is a graph showing the growth inhibitory effect of the human ovarian cancer cell line SK-OV-3 according to the concentration of the administered substance when bougzone ethanol extract alone, doxorubicin alone, and Buchoeson ethanol extract and doxorubicin combination are administered.
FIG. 5 is a graph showing the growth inhibitory effect of human breast cancer cell line MCF-7 according to the concentration of the administered substance when bougzone ethanol extract alone, doxorubicin alone, and bhuozon ethanol extract and doxorubicin combination were administered.
FIG. 6 is a graph showing anticancer efficacy of Buchoeson ethanol extract alone, doxorubicin alone, and Buchoeson ethanol extract and doxorubicin combination in an animal model using a nude mouse.

Little Club Moss is a scientific name of Selaginella Tamariscina (Beauv.) Spring. It is a perennial medicinal plant that grows on rocks distributed throughout Korea. Although basic studies on the anticancer efficacy of Buchoeson have been conducted, research on the use of anticancer adjuvant by combination therapy with clinical anticancer drugs has not yet been conducted. The inventors of the present invention have completed the present invention in view of the fact that the anticancer effect of doxorubicin remarkably increases when doxorubicin, a clinical anticancer agent, and bupjosone ethanol extract are administered concomitantly.

Hereinafter, the present invention will be described in detail.

The present invention is, in one aspect, an anticancer composition comprising an organic solvent extract of Buchucson as an active ingredient.

In the present specification, cancer may mean a state where abnormal tissue growth of body tissue cells results in the formation of a mass and destruction or transformation of the existing structure. Cancer is a tumor in which the growth rate is slow and the tumor is not metastasized Benign tumors and malignant tumors that grow rapidly while invading the surrounding tissues and active in other areas.

In the above aspect, the organic solvent may be alcohol or hexane.

In this aspect, the composition may further comprise a chemotherapeutic agent as an active ingredient.

As used herein, a chemotherapeutic agent means a chemical agent used for the treatment of tumors, for example, to inhibit the proliferation of tumors. The chemotherapeutic agent modifies DNA structure by reacting with N ₇ of guanine of DNA according to the biochemical mechanism of action, and it inhibits the metabolism necessary for the proliferation of alkylating agent and cancer cell, Antibiotics produced by bacteria having antitumor activity, mitotic inhibitors that stop cell division, and hormone agents. The type of the chemotherapeutic agent is not limited as far as it has an anticancer effect.

In the above aspect, the extract of Buchoeson organic solvent, for example, Buchoeson alcohol extract, may be administered in combination with a chemotherapeutic agent.

In the present specification, treatment or amelioration of cancer may mean not only the death of the tumor cells, but also the minimization of the proliferation or the transition from the onset region to another region.

In view of the above, the anti-cancer composition may be used not only for single administration but also for enhancing the anticancer efficacy of a chemotherapeutic agent when it is co-administered with a chemotherapeutic agent. For example, when the composition is administered simultaneously with a chemotherapeutic agent, the anticancer effect of the chemotherapeutic agent can be improved. In one embodiment, the bouchonson ethanol extract has a combination index (CI) value of less than 1 when administered in combination with doxorubicin. The combination index is a numerical value of the synergy effect of the drug, and can be obtained based on the paper "Chou, Theoretical Basis, Experimental Design, and Computerized Simulation of Synergism and Antagonism in Drug Combination Studies, 2006". In one embodiment, the bouchonson ethanol extract has a Combination Index value of less than 1 for solid cancer cells when administered in combination with doxorubicin.

In the above aspect, the bougatons may be at least one selected from the group consisting of roots, stems and leaves of bougatons. I.e., one or more alcohol extracts selected from the group consisting of roots, stems, and leaves of budeson. In one embodiment, the bupcake may be a top portion of a bupchel, and may be, for example, an extract of a stem or leaf of bupchel.

In one aspect of the present invention, the alcohol may be a lower alcohol extract of C ₁ -C ₅ or an aqueous solution thereof.

The concentration of the alcohol aqueous solution may be 10 to 99% (v / v). (V / v) or more, 20% (v / v), 25% (v / v), 50% (v / v), 55% (v / v) or more, 65% (v / v) or more, 70% (v / v) or more, 75% (v / v) or more, 80% (V / v) or higher, 90% (v / v) or higher, 95% (v / v) or higher, or 99% , Not more than 90% (v / v), not more than 85% (v / v), not more than 80% (v / v), not more than 75% (v / v / v), 60% (v / v), 55% (v / v), 50% (v / v), 45% (v / v), or 40% Alcohol aqueous solution, but is not limited thereto. V / v), 60 to 99% (v / v), 70 to 99% (v / v), 70 to 98% (v / v) v), 70 to 97% (v / v), 80 to 99% (v / v), 90 to 99% (v / v), 91 to 99% v / v), 94 to 97% (v / v), 94 to 99% (v / v), 94 to 98% v).

In this respect, the alcohol may be ethanol or methanol.

In the present specification, the term 'extract' includes all substances obtained by extracting the components of a natural product, regardless of the extraction method, extraction solvent, extracted components or extracts, and extracts a substance obtained from the natural product, It is a broad concept that includes all of the materials that can be obtained by processing or processing in a process. For example, a substance obtained through an extraction method including a step of treating with heat, an acid, a base, an enzyme, and the like in the process of extracting a component.

There is no particular limitation on the method for producing burshonse extract used in the present specification. In the present specification, the bushsons extract is a brewer's extract obtained by leaching out boughezhon, transferring the extract, concentrating the brewer's liquid partially or totally, , Or a chemical substance itself exhibiting the main effect contained in the extract.

As used herein, bupchet hands may be included in the form of an extract, or may be included as a pulverized product of the herbal medicine itself or as dry pulverized product of a herbal medicine, but the present invention is not limited thereto. The bougatoon hand used in the present invention is not limited to the method of obtaining the bougainth hand, and the bougie hand may be cultivated and used or purchased commercially.

In one aspect of the present disclosure, the bouchon extract comprises washing and drying the bouchon; Solvent extraction of dried bouganson; And filtering out the extracted bougaintheses. The solvent may include at least one selected from the group consisting of an organic solvent and a mixture of water and an organic solvent. Wherein the water comprises distilled or purified water and the organic solvent is selected from the group consisting of alcohols such as C ₁ -C ₅ lower alcohols, acetone, ether, ethyl acetate, diethyl ether, methanol, ethyl methyl ketone and chloroform , ≪ / RTI > but is not limited thereto. In one embodiment, the alcohol may be ethanol or alcohol.

When bouchetine is extracted with a solvent, it is preferable to extract by adding a solvent corresponding to about 1 to 15 times of bouchetine. Specifically, it is preferable to extract by adding about 10 times of a solvent, but not limited thereto. The extraction may be performed by heat extraction, cold extraction, reflux cooling extraction, ultrasonic extraction or the like, and there is no limitation as long as it is an extraction method that is obvious to a person skilled in the art. The extraction may be carried out at room temperature, but it may be carried out under heating at a temperature of about 40 to 100 ° C, more preferably about 80 ° C, for a more efficient extraction, It is not. The extraction time is preferably about 2 to 4 hours, more preferably about 3 hours, but it is not limited thereto and may be varied depending on conditions such as extraction solvent and extraction temperature. The extraction may be carried out one or more times several times to obtain a larger amount of the active ingredient, preferably 1 to 5 times, more preferably 3 times of continuous extraction.

In addition, in the extraction process of the bouchons extract of the present invention, for example, the organic solvent is added to the fine and dried bougchons, the solution is refluxed and extracted, and the residue and the filtrate are separated by filtration and centrifugation through a filter cloth. And concentrated under reduced pressure to obtain a bouchetto extract. Considering the safety of raw materials, 70% ethanol or butylene glycol is preferably used. In this case, the extraction temperature is preferably 18 to 40 ° C in the case of 70% ethanol and butylene glycol, and the extraction time is preferably 6 to 168 hours. If the extraction temperature and the extraction time are exceeded, the extraction efficiency may be deteriorated or the component may be changed. After obtaining the extract using a solvent, the extract can be obtained by cooling, heating and filtering at room temperature by a conventional method known in the art, or the solvent can be further evaporated, spray dried or freeze-dried. In lyophilization, lyophilization can be performed at -50 to -70 ° C for 3 to 4 days.

In the present specification, the bugotson extract may contain crude extract of bugoseone as described above, and may be included as a soluble fraction of the organic solvent obtained by further extracting the crude extract with an organic solvent having a low polarity. Examples of the organic solvent include, but are not limited to, hexane, methylene chloride, ethyl acetate, n-butanol, and the like. The extract or the soluble fraction of the extract may be used as it is, or may be used as an extract form by concentration after filtration, and may be used as a form of a lyophilizate by concentration and freeze-drying.

The composition of the extract may vary depending on the extraction solvent. For example, in high-performance liquid chromatography (HPLC) analysis, the peach appearance of the Buchoeson ethanol extract or Buchoeson methanol extract is significantly different from the Buchoeson water extract.

In this aspect, the cancer may be a solid cancer or a blood cancer. In the present specification, solid cancer may mean cancer that occurs in solid organs such as stomach cancer, liver cancer, lung cancer, breast cancer, ovarian cancer, thyroid cancer, pancreatic cancer, colon cancer and the like. Solid tumors are different in non-solid cancer (also called blood cancer) such as leukemia, cancer site, cancer metastasis method and treatment method.

In the above aspect, the solid cancer may include lung cancer, ovarian cancer, breast cancer, liver cancer, rectal cancer, gallbladder cancer, cholangiocarcinoma, colon cancer, pancreatic cancer, bronchial cancer, bladder cancer, soft tissue osteosarcoma or brain tumor.

Generally, lung cancer is divided into non-small cell lung cancer and small cell lung cancer depending on the size and shape of cancer cells. The non-small cell lung cancer is divided into a squamous cell carcinoma arising from the metaplasia of the squamous epithelium, constituting cells of the bronchial mucosa of the lung, and adenocarcinoma arising from the lung adenocarcinoma and a large cell carcinoma of large cancer cell size.

In the above aspect, the lung cancer may be adenocarcinoma or large cell cancer.

In this aspect, the blood cancer may include leukemia or malignant lymphoma.

In the anticancer composition according to one aspect of the present invention, the chemotherapeutic agent may include an anthracycline compound. An anthracycline-based chemotherapeutic agent is a kind of alkylating agent and is mainly used for actinomycetes ( Streptomyces There is peucetius . may be a group of antibiotics that have anticancer effects produced by cucumber seedlings. For example, the antracycline antibiotics may be daunomycin, adriamycin, doxorubicin, acclinomycin A or salts thereof. Doxorubicin has religiously ^®, Maio set ^®, As Carrick's ^® may be one made of a collection form liposomes (liposome-encapsulated forms). In one embodiment, the chemotherapeutic agent may be an anthracycline based compound and may be doxorubicin.

Doxorubicin may be a substance of the formula C ₂₇ H ₂₉ NO ₁₁ (Formula I), and the molar mass may be 543.52 g / mol. Toxorubicin can be used in hydrochloride form to enhance water solubility. The doxorubicin hydrochloride may be present in the molecular structure of formula (C ₂₇ H ₂₉ NO ₁₁ HCl), wherein the molar mass may be 579.98 g / mol.

[Chemical Formula 1]

(2)

In the above aspect, the organic solvent extract of Buchucson may be contained in an amount of 0.01 wt% or more by dry weight. For example, the Buchoeson organic solvent extract may contain at least 0.001% by weight, at least 0.01% by weight, at least 0.05% by weight, at least 0.07% by weight, at least 0.1% by weight, at least 0.3% by weight, at least 0.5% by weight, at least 0.7% by weight, At least 3% by weight, at least 5% by weight, at least 7% by weight, at least 9% by weight, at least 10% by weight, at least 15% by weight, at least 20% by weight, at least 30% by weight, at least 40% by weight Or 50% by weight or more, but is not limited thereto. In one embodiment, the organic solvent extract may be an alcohol extract.

In the above aspect, the chemotherapeutic agent and the organic solvent extract of Bucheosan may be contained in the composition for treating or improving cancer in a weight ratio of 1: 1 ~ 300. The organic solvent may be an alcohol or hexane, for example, ethanol.

For example, when doxorubicin is included at concentrations of 0.0125, 0.25, 0.5, 1, 2 and 4 μM, the Buchoeson organic solvent extract, such as Buchoeson's alcohol extract, To 70 and 70 to 200 [mu] g / ml. The weight ratio of the chemotherapeutic agent and the organic solvent extract of Bucheoson, for example, Buchoeson's alcohol extract, in the composition may be 1: 0.1 to 300, 1: 0.1 to 250, 1: 0.1 to 200, 1: 0.1 to 150, 1 to 100, 1 to 100, 1 to 100, 1 to 100, 1 to 100, 1 to 100, 1 to 10, 150, or 1: 10-100.

In one embodiment, the organic solvent may be an alcohol, specifically ethanol.

In this aspect, the composition may be a pharmaceutical composition or a health functional food composition.

The pharmaceutical compositions according to the present disclosure may be of various oral or parenteral formulations. 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 form preparations for oral administration include tablets, pills, powders, granules, soft or hard capsules, etc. These solid preparations may contain one or more excipients such as starch, calcium carbonate, sucrose, Or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The pharmaceutical dosage forms of the compositions herein may be used in the form of their pharmaceutically acceptable salts and may also be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable set. The salt is not particularly limited as long as it is pharmaceutically acceptable so long as it is pharmaceutically acceptable and includes, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, , Benzenesulfonic acid, toluenesulfonic acid, and naphthalenesulfonic acid.

The composition of the present invention may be administered parenterally or orally, and may be administered in one to several divided doses in an amount of 0.1 to 500 mg, preferably 1 to 100 mg per kg of body weight per day have. The dosage for a particular patient may vary depending on the patient's body weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, severity of disease, and the like.

The pharmaceutical composition according to the present invention may be formulated into various forms such as powders, granules, tablets, soft or hard capsules, oral formulations such as suspensions, emulsions, syrups and aerosols, external preparations such as ointments and creams, suppositories, Sterile injectable solutions, and the like, and may be formulated and used in any form suitable for pharmaceutical preparations.

The compositions according to the present disclosure can be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes including parenterally, orally, and all manner of administration can be expected, for example, Or by intravenous, intramuscular, subcutaneous, intramural or intracerebroventricular injection.

In one aspect of the present disclosure, the food composition may be a health functional food composition.

The formulation of the food composition according to the present specification is not particularly limited, but may be formulated into, for example, tablets, granules, powders, liquid preparations such as a drink, caramels, gels, bars and the like. The food composition of each formulation can be blended with the ingredients commonly used in the field in addition to the active ingredient without difficulty by those skilled in the art depending on the purpose of formulation or use, and synergistic effect can be obtained when the composition is applied simultaneously with other ingredients.

In the food composition according to the present specification, the determination of the dosage of the active ingredient is within the level of those skilled in the art, and its daily dosage is, for example, from 0.1 mg / kg / day to 5000 mg / kg / day, The dose may be 50 mg / kg / day to 1000 mg / kg / day, but is not limited thereto, and may vary depending on various factors such as the age, health condition, and complications of the subject.

The food composition according to the present invention may be used as a food or beverage such as various foods such as chewing gum, caramel product, candy, ice cream, confectionery, beverage such as soft drink, mineral water, alcoholic beverage, healthful food including vitamins and minerals .

In addition to the above, the food composition which is one aspect of the present invention may be in the form of a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the functional food compositions of the present disclosure may include natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally comprised in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition herein.

In another aspect, the present invention includes a composition comprising the bougue's organic solvent extract, a composition comprising a chemotherapeutic agent, and an instruction sheet, wherein the bougue's organic solvent extract and the chemotherapeutic agent are mixed at a weight ratio of 1: In combination with the administration of the anti-cancer kit.

In the above-mentioned aspect, the instructions may include the co-administration of the organic solvent extract of Bukhandan and the chemotherapeutic agent at a weight ratio of 1:10 to 100. The weight ratio is not limited and may be 1: 0.1 to 300, and may be 1: 0.1 to 250, 1: 0.1 to 200, 1: 0.1 to 150, 1: 0.1 to 130, 1: 0.1 to 100, , 1: 1 to 200, 1: 1 to 150, 1: 1 to 100, 1: 10 to 300, 1:10 to 200, 1:10 to 150, or 1:10 to 100.

In one embodiment, the organic solvent may be an alcohol, specifically ethanol.

In one aspect of the present invention, the chemotherapeutic agent may comprise an anthracycline compound. In one embodiment, the anthracycline compound may be doxorubicin.

Hereinafter, the present invention will be described in more detail with reference to the following examples and experimental examples. However, these examples and experimental examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited by the following examples.

[ Example  1] Preparation of Buchoeson Ethanol Extract

The dried bruchon, which was purchased on the market, was pulverized to an appropriate size, and 200 g of Buchoes and 3.0 L of a 95% aqueous ethanol solution were added to the extraction vessel. The mixture was refluxed and filtered to obtain an extract. The extraction procedure was repeated three times, and then the solvent was concentrated under reduced pressure to obtain 24.5 g of an ethanol extract.

[ Example  2] Preparation of Bucheoson water extract

Dried budsons were purchased from the market and pulverized to an appropriate size. 10 g of Bucherson and 200 ml of distilled water were added to the extraction vessel, refluxed, and filtered to obtain an extract. The extraction procedure was carried out once, after which the solvent was concentrated under reduced pressure and lyophilized to obtain 0.827 g of water extract. In addition, 50 mg of the water extract prepared according to need was autoclaved at 121 ° C for 15 minutes using a high-pressure sterilizer (TOMY SX-700).

[ Example  3] Preparation of methanol extract of Buchoeson

100 g of Bucherson and 200 ml of methanol were added to the extraction vessel, and the mixture was refluxed. The extract was obtained by filtration through filter paper. The extraction procedure was performed once, and then the solvent was concentrated under reduced pressure to obtain 1.32 g of a methanol extract.

[ Example  4] High Performance Liquid Chromatographic Analysis of Bucherson Extracts

High - performance liquid chromatography (HPLC) analysis was performed to compare the chemical composition of bouchet extract. The instrument used for the analysis was a 1260 model of Agilent, and MG-II reverse phase column (4.6Φ × 250 mm, 5 μm) of Shiseido Co. was used as a stationary phase, 0.1% trifluoroacetic acid was added to water and acetonitrile And analyzed at a flow rate of 1 ml per minute. The changes in the mobile phase composition with time were as shown in Table 1, and the absorbance was measured at 254 nm in ultraviolet light.

Change of mobile phase composition Time (min) Water (%) Acetonitrile (%) 0 80 20 40 10 90 45 10 90

HPLC analysis showed that the peaks of ethanol extracts and methanol extracts showed similar peaks, while those of water extracts were significantly different from those of methanol extracts. When the water extracts were sterilized under high pressure, I could see that it disappeared. Therefore, the composition of Buchoeson ethanol extract and methanol extract are similar to each other, but the composition of Buchoeson alcohol (ethanol or methanol) and Buchoeson water extract are completely different.

[ Test Example  1] Buchoeson ethanol extract and Water extract  Measurement of the inhibitory activity against cancer cell growth on human lung cancer cell line A549 and measurement of cell survival rate in combination with doxorubicin

The cell survival rate of A549 cell line (Korean Cell Line Bank), which is a human lung cancer cell line, was measured while the Buchoeson ethanol extract of Example 1 or the Buchoeson water extract of Example 2 was treated alone or in combination with doxorubicin. A549 cells were cultured in RPMI medium (Roswell Park Memorial Institute medium) supplemented with 10% fetal bovine serum (FBS), 10 U / ml penicillin and 100 μg / ml streptomycin, Lt; / RTI > The medium solution and the A549 cell culture solution were mixed, and ³ x 5 x 10 cells were inoculated into a 96-well plate and cultured for 24 hours at 5% carbon dioxide and 37 ° C. After the cells were stabilized, the Buchoeson ethanol extract and the water extract were individually treated at a concentration of between 3.125 and 100 占 퐂 / ml with a 2-fold concentration gradient, or the doxorubicin was treated with a concentration of between 0.125 and 4 占 2 Or bupcahson extract and doxorubicin, followed by incubation for 24 hours at 5% carbon dioxide and 37 ° C. Cell viability was then measured. The cell viability was measured by adding 450 μl of EZCytox reagent (Daeil Lab Service, Korea) and incubating the cells at 5% carbon dioxide and 37 ° C for 1-4 hours. The cell viability was measured by dimethylsulfoxide (DMSO ) Was calculated as the survival rate and the ratio of the control group.

As a result, it was found that Buchoeson water extract did not significantly inhibit the growth of lung cancer cells, whereas Buchoeson ethanol extract inhibited the growth of lung cancer cells in a dose dependent manner. In addition, when Buchoeson ethanol extract was administered in combination with doxorubicin, the survival rate of lung cancer cells was further lowered than when doxorubicin was administered alone, and it was confirmed that Buchoeson ethanol extract had a synergistic effect with doxorubicin. On the other hand, it was found that Buchoeson water extract inhibited the anticancer efficacy of doxorubicin in combination with doxorubicin (Fig. 1A, Fig. 2).

[ Test Example  2] Human lung cancer cell line of Buchoeson ethanol extract NCI- H460  And the measurement of cell survival rate in combination with doxorubicin

The cell survival rate of the human lung cancer cell line NCI-H460 cell line (Korean Cell Line Bank) was measured while the Buchoes' ethanol extract of Example 1 was treated alone or in combination with doxorubicin. NCI-H460 cells were cultured in RPMI medium (Roswell Park Memorial Institute medium) supplemented with 10% fetal bovine serum (FBS), 10 U / ml penicillin and 100 μg / ml streptomycin, Lt; 0 > C. The medium solution and NCI-H460 cell culture medium were mixed, and 5 x 10 ³ cells were inoculated into a 96-well plate and cultured for 24 hours at 5% carbon dioxide and 37 ° C. After the cells are stabilized, Buchoeson ethanol extract is treated with a concentration of between 3.125 and 100 / / ml in a 2-fold concentration gradient, or with doxorubicin alone at a concentration between 0.0625 and 2 쨉 M in a 2-fold concentration gradient, And doxorubicin, and then cultured for 24 hours at 5% carbon dioxide and 37 ° C. Cell viability was then measured. The cell viability was measured at 450 nm after the addition of EZCytox reagent (Daeil Lab Service, Korea), followed by incubation for 1 to 4 hours at 5% carbon dioxide and 37 ° C. The cell viability was measured using a DMSO- Of survival rates and ratios.

As a result, it was found that Buchoeson ethanol extract inhibited the growth of lung cancer cells in a concentration - dependent manner. In addition, when bouchones ethanol extract was administered in combination with doxorubicin, the survival rate of lung cancer cells was further lowered than when doxorubicin alone was administered, and it was confirmed that bouchones ethanol extract had a synergistic effect with doxorubicin (Fig. 3).

[ Test Example  3] Human of bouchet ethanol extract Ovary cancer cell line  SK- OV -3 and measurement of cell survival rate in combination with doxorubicin

The cell survival rate of the SK-OV-3 cell line (Korean Cell Line Bank), which is a human ovarian cancer cell line, was measured in the same manner as in Test Example 2. [ As a result, Buchoeson ethanol extract inhibited the growth of ovarian cancer cells in a dose - dependent manner. In addition, when Buchoeson ethanol extract was administered in combination with doxorubicin, the survival rate of ovarian cancer cells was further lowered compared to when doxorubicin alone was administered, and it was confirmed that Buchoeson ethanol extract had a synergistic effect with doxorubicin against ovarian cancer cells 4).

[ Test Example  4] Human of bouchet ethanol extract Breast cancer cell line MCF -7 in cancer cell growth inhibition assay and measurement of cell survival rate in combination with doxorubicin

The cell survival rate of the human breast cancer cell line MCF-7 (Korean Cell Line Bank) was measured in the same manner as in Test Example 2. As a result, it was found that Buchoeson ethanol extract inhibited the growth of breast cancer cells in a concentration-dependent manner. In addition, when bouchones ethanol extract was administered in combination with doxorubicin, the survival rate of breast cancer cells was further lowered than when doxorubicin alone was administered, and it was confirmed that bouchones ethanol extract had synergy with doxorubicin against breast cancer cells (FIG. 5) .

[ Test Example  5] Synergistic or antagonistic effect of anticancer effect when combined with bouczone extract and doxorubicin

When Buchoeson's ethanol extract of Example 1 or Buchoeson's water extract of Example 2 is administered in combination with doxorubicin, a clinical anticancer agent, whether or not bupchezson extract induces synergism that raises the anticancer efficacy of doxorubicin, or whether doxorubicin Combination index (CI) was calculated in order to confirm antagonism that would interfere with anticancer efficacy. Combination indices were calculated by reference to the paper (Chou, Theoretical Basis, Experimental Design, and Computerized Simulation of Synergism and Antagonism in Drug Combination Studies, 2006). COMPUSYN software (version 1.0, ComboSyn) was used for this purpose. Generally, when the combination index is 1 or less, the synergistic effect is exhibited. If the combination index is greater than 1, the synergistic effect is exhibited. Table 2 shows the criteria for the synergistic or antagonistic action according to the combination index value.

Combination index Explanation ≪ 0.1 Very strong synergism 0.1 - 0.3 Strong synergism 0.3 - 0.7 Synergism 0.7 - 0.85 Moderate synergism 0.85 - 0.90 Slight synergism 0.90 - 1.10 Nearly additive 1.10 - 1.20 Slight antagonism 1.20 - 1.45 Moderate antagonism 1.45 - 3.3 Antagonism 3.3 - 10 Strong antagonism > 10 Very strong antagonism

As a result, bucheoson water extract against human lung cancer cell line A549 is a combination index values ED ₅₀ is _1.88, ED ₇₅ was 8.31. In other words, it was confirmed that Bucheosone water extract inhibited the anticancer effect of doxorubicin by antagonistic action with doxorubicin against human lung cancer cell line.

As a result of measurement of the combination index using the Buchoeson ethanol extract, the ED ₅₀ was 0.37 _{and the} ED ₇₅ was 0.44 for the human lung cancer cell line A549. For the human lung cancer cell line NCI-H460, the ED ₅₀ was 0.68 _{and the} ED ₇₅ was 0.62 . For the human ovarian cancer cell line SK-OV-3, the ED ₅₀ was 0.54 _{and the} ED ₇₅ was 0.70. For the human breast cancer cell line MCF-7, the ED ₅₀ was 0.80 _{and the} ED ₇₅ was 0.88. That is, the Buchoeson ethanol extract had a combined index value of less than 1 for lung cancer, ovarian cancer, and breast cancer cell lines, indicating that Buchoeson ethanol extract synergizes with the anticancer efficacy of doxorubicin. In other words, it was confirmed that Buchoeson ethanol extract maximizes the anticancer effect against doxorubicin lung cancer, ovarian cancer or solid cancer such as breast cancer.

[ Test Example  6] Human Lung cancer cell line  A549 BALB / c Inhibition of Tumor Growth upon Administration of Buchoeson Ethanol Extract and Doxorubicin in a Xenotransplantation Mouse Model Introduced into Nude Mouse

Human lung cancer cell line A549 was cultured in RPMI medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin, and streptomycin (Roswell Park Memorial Institute medium) under the condition of 5% Respectively. The cultured cell lines were removed and mixed in RPMI medium containing no fetal bovine serum. Then, female BALB / c nude mice at ⁷ weeks were injected into the left side of the mouse at a concentration and volume of 1 × 10 7/200 μl, respectively Animals were raised for 2 weeks and the tumor volume was measured. The amount of tumor was calculated by the following formula: tumor volume = (width × height ² ) / 2 (mm ³ ). Two weeks later, when the amount of tumors in each mouse reached about 200 mm ³ , bupjosone ethanol extract and doxorubicin combination therapy were administered. Experimental groups were divided into four groups: PBS (phosphate buffered saline) supplemented with 1% DMSO (dimethyl sulfoxide), intraperitoneally injected with PBS (group 1, control group), Buchoeson ethanol extract was added with 1% DMSO PBS was administered intraperitoneally (group 2, bupsellone extract alone) or oral administration of PBS supplemented with 1% DMSO at a concentration of 1 mg / kg of PBS at a concentration of 1000 mg / kg. After diluting with PBS, intraperitoneal injection (Group 3, doxorubicin alone), and finally, Buchoeson ethanol extract was dissolved in PBS containing 1% DMSO at a concentration of 1000 mg / kg, followed by oral administration of doxorubicin at 1 mg / kg (Group 4, bupcitabine extract and doxorubicin combination group), and then administered for 26 days, and the amount of tumor was measured. Because the initial tumor volume that started to be used in combination was different for each mouse, the relative value of the increased tumor volume based on the size of the tumor on the day of initiation of the combination therapy was compared.

As a result, it was confirmed that the cancer cell growth of the mice treated with Buchoeson ethanol extract and doxorubicin alone was inhibited. Further, when the Buchoeson ethanol extract and doxorubicin were administered together, doxorubicin and bouchones ethanol extracts were administered alone The cancer cell growth was further suppressed, and it was confirmed that the Buchoeson ethanol extract had a synergistic effect of increasing the anticancer efficacy of doxorubicin (Fig. 6).

Formulation examples of compositions according to one aspect of the present disclosure are described below, but are also applicable to various other formulations, which are intended to be illustrative only and not for purposes of limitation.

 [Formulation Example 1-1] Soft capsule 1

The soft capsule filling liquid is prepared by mixing 0.1 mg of Buchoes' ethanol extract of Example 1, 9 mg of vitamin E, 9 mg of vitamin C, 2 mg of palm oil, 8 mg of vegetable hardening oil, 4 mg of yellow pear and lecithin of 9 mg according to a conventional method. 400 mg per capsule is filled to prepare a soft capsule. Separately from the above, a soft capsule sheet was prepared in a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin and 10 parts by weight of sorbitol solution, and filled with the filling solution to prepare a soft capsule containing 400 mg of the composition according to the present invention.

[Formulation Example 1-2] Soft capsule 2

The soft capsule filling solution was prepared by mixing 0.1 mg of Buchoeson ethanol extract of Example 1, 10 mg of doxorubicin, 9 mg of vitamin E, 9 mg of vitamin C, 2 mg of palm oil, 8 mg of vegetable hardened oil, 4 mg of yellow pearls and 9 mg of lecithin and mixing them according to a usual method do. 400 mg per capsule is filled to prepare a soft capsule. Separately from the above, a soft capsule sheet was prepared in a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin and 10 parts by weight of sorbitol solution, and filled with the filling solution to prepare a soft capsule containing 400 mg of the composition according to the present invention.

[Formulation Example 2] Tablets

A mixture of 0.1 mg of Buchoes' ethanol extract of Example 1, 9 mg of vitamin E, 9 mg of vitamin C, 200 mg of galactooligosaccharide, 60 mg of lactose and 140 mg of maltose was granulated using a fluid bed drier, and 6 mg of sugar ester Lt; / RTI > 500 mg of these compositions are tabletted by conventional methods to prepare tablets.

[Formulation Example 3] Drinking agent

0.075 mg of Buchoes' ethanol extract of Example 1, 9 mg of vitamin E, 9 mg of vitamin C, 10 g of glucose, 0.6 g of citric acid and 25 g of liquid oligosaccharide are mixed and 300 ml of purified water is added to fill each bottle 200 ml each. It is filled in a bottle and sterilized at 130 ° C for 4 to 5 seconds to prepare a drink.

[Formulation Example 4]

0.1 mg of Buchoes' ethanol extract of Example 1, 9 mg of vitamin E, 9 mg of vitamin C, 250 mg of anhydrous crystalline glucose and 550 mg of starch were mixed and granulated into granules using a fluidized bed granulator.

[Formulation Example 5]

Injections were prepared in a conventional manner according to the composition shown in Table 3 below.

Compounding ingredient content The bouchet ethanol extract of Example 1 0.01-0.05 mg Sterile sterilized water for injection Suitable amount pH adjusting agent Suitable amount

[Formulation Example 6] Health functional food

Health functional foods were prepared according to the compositions shown in Table 4 below in a conventional manner.

Compounding ingredient content The bouchet ethanol extract of Example 1 75 쨉 g Vitamin A acetate 70 g Vitamin E 1.0 mg Vitamin B1 0.13 mg Vitamin B2 0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 g Vitamin C 10 mg Biotin 10 g Nicotinic acid amide 1.7 mg Folic acid 50 μg Calcium pantothenate 0.5 mg Ferrous sulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium monophosphate 15 mg Dicalcium phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a component suitable for a health functional food as a preferred embodiment, the compounding ratio may be arbitrarily modified.

[Formulation Example 7] Health drinks

Health drinks were prepared in a conventional manner according to the composition shown in Table 5 below.

Compounding ingredient content The bouchet ethanol extract of Example 1 50 μg Citric acid 1000 mg oligosaccharide 100 g Taurine 1g Purified water Balance

The above components are mixed according to a conventional health drink manufacturing method, and the mixture is stirred and heated at 85 DEG C for about 1 hour, and then the solution is sterilized by filtration.

Having described specific portions of the disclosure in detail, those skilled in the art will appreciate that these specific embodiments are merely exemplary and are not intended to limit the scope of the disclosure. Accordingly, the actual scope of the disclosure will be defined by the appended claims and their equivalents.

Claims (23)

An organic solvent extract of Buchoesan and a chemotherapeutic agent as active ingredients,
Wherein the chemotherapeutic agent is doxorubicin. The method according to claim 1,
Wherein the organic solvent is an alcohol or hexane. delete delete The method according to claim 1,
Wherein the budsonza is a topical part of budeson. 6. The method of claim 5,
The anticancer composition according to claim 1, wherein the root portion of the boucheon is a leaf or stem of bouchet. 3. The method of claim 2,
Wherein the alcohol is a C ₁ -C ₅ lower alcohol or an aqueous solution thereof. 8. The method of claim 7,
Wherein the concentration of the aqueous alcohol solution is 10 to 99% (v / v). 8. The method of claim 7,
Wherein the alcohol is ethanol or methanol. The method according to claim 1,
Wherein the cancer is solid cancer. 11. The method of claim 10,
The solid-
Lung cancer, ovarian cancer, or breast cancer. 12. The method of claim 11,
Wherein said lung cancer is non-small cell lung cancer. 13. The method of claim 12,
Wherein said non-small cell lung cancer is adenocarcinoma or large cell carcinoma. delete delete The method according to claim 1,
The anticancer composition according to claim 1, wherein the bougouzone organic solvent extract is contained in an amount of 0.01 wt% or more by dry weight. The method according to claim 1,
The chemotherapeutic agent and the organic solvent extract of Bucheosan,
1: 1 to 300, by weight. The method of claim 1, wherein
The chemotherapeutic agent and the organic solvent extract of Bucheosan,
1: 10-100. ≪ / RTI > The method according to claim 1,
The composition may comprise,
Wherein the composition is a pharmaceutical composition or a health functional food composition. A composition comprising Buchoeson organic solvent extract;
A composition comprising doxorubicin as a chemotherapeutic agent; And
Includes instructions,
Wherein the directive contains the step of administering the bougainza organic solvent extract and the chemotherapeutic agent in a weight ratio of 1: 1 to 300 in combination. 21. The method of claim 20,
Wherein said instructions include the simultaneous administration of Bucheon's organic solvent extract and chemotherapeutic agent at a weight ratio of 1:10 to 100. delete delete

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