KR101283562B1 - Composition for inhibition of multidrug resistance containing an extract of Trichosanthes kirilowii maxim - Google Patents

Abstract

The present invention provides a pharmaceutical composition for inhibiting multi-drug resistance, comprising as an active ingredient a cheonhwamin extract excellent in multidrug resistance (MDR) inhibitory ability. The composition of the present invention can overcome the anticancer drug resistance in multidrug resistant cells showing tolerance to an anticancer drug and can be used as a pharmaceutical composition and anticancer adjuvant for inhibiting multiple drug resistance.

Description

Composition for inhibition of multidrug resistance containing an extract of Trichosanthes kirilowii maxim}

The present invention is very effective in inhibiting multidrug resistance (MDR) ( trichosanthes) kirilowii maxim) relates to a composition for inhibiting multi-drug resistance comprising an extract as an active ingredient.

To date, various treatments such as surgical, radiation, and chemotherapy have been studied to treat cancer, but they have not yet been fully treated.

Of these, surgery and radiation therapy are useful in the early stages of cancer development. Therefore, in the case of cancer, which is difficult to detect early on, chemotherapy is increasingly relied upon and chemotherapy has a history of more than 50 years Hundreds of anticancer drugs have been developed and used in clinical practice. However, there are still few cases in which clinically satisfactory effects are obtained.

Chemotherapy is well known as one of the most effective methods of treating cancer patients, but continuous administration of chemotherapy has been pointed out as a cause of failure of chemotherapy. This is because the cells acquire resistance to the anticancer drug, and it is known that acquiring resistance to an anticancer drug also acquires resistance against different anticancer drugs. The fact that cancer cells acquire nonspecific resistance regardless of the anticancer structure is called "Multidrug Resistance (MDR)" (Gottesman et al., Nat Rev Cancer 2: 48-58, 2002 Ambudkar et al., Oncogene 22: 7468-85, 2003). MDR acquisition of these cancer cells is well known as one of the major causes of failure of chemotherapy. The anticancer drug resistance can be categorized into two types: one is caused by the inability to enter the cells into the drug, and the other is that the cancer cells themselves are susceptible to anticancer drugs by genetic or epigenetic changes Can fall apart.

The mechanisms by which cancer cells acquire MDRs can be divided into five broad categories. First, the efflux pump such as ATP-dependent transporter can be used to easily extrude the intracellular drug out of the cell to maintain the concentration lower than the intracellular effective drug concentration to induce tolerance. Secondly, the drug transporter or endo Or the drug resistance is induced by promoting / inactivation of the metabolism of the drug introduced into the cell, or, fourth, the DNA caused by the drug (NF-kB signaling pathway activation), which can inhibit drug-induced apoptosis, and (5) inhibit the drug-induced apoptosis. Ceramide concentration reduction, etc.) can be induced.

Among them, MDR acquisition of cancer cells by drug overexpression overexpression causes problems of high dose of anticancer drug for effective treatment and may cause bone marrow toxicity which is the biggest problem of existing anticancer drug. In other words, when the cancer cell is resistant to overexpression by the overflow pump, the cancer cell can effectively survive by leaking the anticancer agent to the outside of the cell. However, since the expression of the effusion pump is low in the bone marrow, And bone marrow toxicity may be increased. Therefore, resistance to outflow pump overexpression is the most serious in resistance expression, and in order to overcome cancer successfully by using chemotherapy, it is possible to effectively control the outflow pump which can overcome it, (Gottesman et al., Nat Rev Cancer 2: 48-58, 2002; Friedenberg et al., Cancer 106: 830-8, 2006; Seiden et al., Gynecol Oncol 86: 302-10, 2002).

The drug efflux pump is a membrane protein belonging to the ATP-binding cassette (ABC) transporter family when comparing the amino acid sequence and structural characteristics. Up to now, 48 ABC genes have been identified in humans and are divided into 7 subfamilies (ABCA-ABCG). Among them, MDR1, multidrug-resistant protein (MRP) 1-7, BSEP / SPGP, and MXR / BCRP / ABC-P are examples of ABC transporters involved in drug transport. The drug-transporting ABC transporter plays a role in non-specifically injecting the intracellular drug into the cell regardless of the structure of the drug, which is known to be most important for MDR expression (Endicott et al., Annu Rev Biochem 58 : 137-71, 1989; Ambudkar et al., Annu Rev Pharmacol Toxicol 39: 361-98, 1999; Gottesman et al., Nat Rev Cancer 2: 48-58, 2002).

Drug transport The most well-known of the ABC transporters is the P-glycoprotein (P-gp), the product of the ABCB1 (or MDR1) gene. P-gp is a membrane protein with 12 transmembrane domains of 170 KDa and two ATP binding sites. It is the most abundant anti-cancer drug (vinca alkaloids, taxol, etoposide, And the like) as a substrate. When a hydrophobic drug (neutral or cationic drug) binds to the transmembrane domain of P-gp, the hydrolysis of the two molecules leads to a morphological change in P-gp leading to the release of the drug outside the cell (Ambudkar et al ., Annu Rev Pharmacol Toxicol 39: 361-98, 1999). Thus, broad substrate specificity of P-gp is believed to be due to polymorphism that can bind to various drugs in the transmembrane domain, the drug binding site. In addition, MRP1 (multidrug-resistance-associated protein 1; ABCC1) is also important for MDR expression. MRP1 is primarily responsible for the binding and release of neutral or anionic hydrophobic drugs and drugs associated with glutathione (Gottesman et al., Nat Rev Cancer 2: 48-58, 2002).

The increase in P-gp expression in cancer cells after treatment with chemotherapy is largely explained by two models. First, the model was proposed about 20 years ago as a "selection and expansion model." In other words, when cancer develops, P-gp-expressing cells and less-expressing cells coexist. In the course of chemotherapeutic treatment, cells that express less P-gp die, (Goldie & Coldman, Cancer Res 44: 3643-53, 1984). In the present study, we examined the effect of P-gp overexpressing cells on the proliferation of P-gp overexpressing cells. The second model is an "activation model" in which the chemotherapeutic agent directly activates the MDR1 gene to overexpress P-gp and thereby express drug resistance (Abolhoda et al., Clin Cancer Res 19995: 3352-6, 1999).

Recently, we have developed a substance that can inhibit the action of a discharge pump such as P-gp in order to enhance the cancer treatment effect. By using this drug together with existing anticancer drugs, it is possible to reduce resistance to anticancer drugs and decrease the dose of anticancer drugs. Researches are being actively carried out. These resistant inhibitors are called MDR reversing agents, chemosensitizers or multidrug resistance modulators (MDR modulators). The first developed multidrug resistance reversing agent is verapamil, which is a P-gp inhibitor. Since then, various substances have been developed and used, but other side effects due to cytotoxicity have been caused. In addition, as for the patent on multidrug resistance, the imidazole compound (Korean Patent No. 0330698, 0349500), chromone compound (Korean Patent No. 0580743, Korean Patent Publication No. 2005-0034024), quinoline compound (Korean Patent Laid-Open Publication No. 1998-7000978, 1998-700972), Korean native plants such as Sangbangcho, Soryeongdae, (Korean Patent Laid-Open No. 2006-0124453), a vaccine against P-glycoprotein 170 for MDR inhibition (Korea Patent Publication No. 2006-0125676), and an antibody against MDR product (Korean Patent Publication No. 1995-7000938).

Fern on hand ( Trichosanthes kirilowii maxim refers to the peeled root of the gourd (Trichosanthes kirilowii Maximowicz) or Yellow Hanultari (Trichosanthes kirilowii Max. var. japonica Kitamura). Hanultari is a vine plant, and the Guara (果?), Which combines the fruit on the tree and the fruit on the ground, is said to have become a lump (루). When the roots of this herb are made of powder, they are clean as snow, so it is called Cheonhwabun. Appearance uneven columnar, sometimes divided longitudinally. The outer surface is light yellowish white with yellowish brown irregular tube extending. Folded cotton pale yellow, slightly fibrous. Other names include Fruit Root, Fruit Root, Gwalup Root, Gwalupbun, Lubun, White Medicine, Seoseo And Lugeun.

Cranberries treat thirst, boils and pus when the fluid is damaged by heat. Mainly lowers the heat of the lungs and stomach, making the essences to quench thirst and soothe the body. Pharmacological actions have been reported to inhibit malignant grapes and epithelial cell carcinoma, uterine smooth muscle excitability, hepatocarcinogenic effect, hypoglycemic activity, and bactericidal action. However, the results of research as a multidrug resistance inhibitor as a physiological activity of hwahwahwa have not been reported.

Accordingly, the present inventors while studying a new multi-drug resistance inhibiting composition from natural products (plants and herbs) in order to overcome the MDR phenomenon which is the biggest failure of chemotherapy and prevent other side effects due to cytotoxicity, The present invention was completed by confirming that the extract inhibits P-glycoprotein expression and inhibits the expression of the MDR1 gene to increase the sensitivity to anticancer drugs in multi-drug resistant cancer cell lines.

It is an object of the present invention to provide a pharmaceutical composition for inhibiting multidrug resistance which increases the sensitivity to an anticancer drug in a multidrug-resistant cancer cell line in order to overcome the multidrug resistance which is a problem of chemotherapy.

It is another object of the present invention to provide a pharmaceutical composition for assisting an anticancer agent which increases sensitivity to an anticancer drug in a multidrug-resistant cancer cell line in order to overcome multiple drug resistance which is a problem of chemotherapy.

It is another object of the present invention to provide an anticancer drug-supplementing food composition for enhancing sensitivity to an anticancer drug in a multidrug-resistant cancer cell line to overcome multiple drug resistance, which is a problem of chemotherapy.

In order to solve the above problems, the present invention provides a composition for inhibiting multi-drug resistance comprising a cheonhwamin extract as an active ingredient.

The present invention also provides an anticancer adjuvant pharmaceutical composition comprising a cheonhwamin extract as an active ingredient.

In another aspect, the present invention provides anti-cancer auxiliary food composition comprising a natural extract as an active ingredient.

The composition of the present invention has excellent antimicrobial resistance-inhibiting activity by P-glycoprotein in multidrug resistance (MDR) cells showing tolerance to an anticancer drug, so that the treatment of cancer patients with anticancer drug resistance ≪ / RTI >

FIG. 1 is a graph showing the expression of MDR1 in the KB cell line, which is a normal cancer cell, KB / VB20 cell line, which is a multidrug resistant cancer cell, and A549 cell line and Caco-2 cell line, respectively, as a control group.
Figure 2 shows the results of Rhodamine 123 assay of cheonhwabun extract.
Figure 3 is a figure analyzing the effect of cheonhwamin extract on the expression of P-glycoprotein.
Figure 4 is a figure analyzing the effect of cheonhwamin extract on the gene expression of MDR1.
Figure 5 is a figure analyzing the effect of cheonhwamin extract on YB-1 gene expression.
FIG. 6 is a graph illustrating the effect of cheonhwamin extract on the sensitivity of vinblastine, an anticancer agent, in multi-drug resistant cancer cells.

Hereinafter, the present invention will be described in more detail.

The present invention provides a composition for inhibiting multi-drug resistance, comprising a natural extract of cheonhwabun as an active ingredient.

The present invention also provides an anticancer adjuvant pharmaceutical composition comprising a cheonhwamin extract as an active ingredient.

In another aspect, the present invention provides anti-cancer auxiliary food composition comprising a natural extract as an active ingredient.

The natural extract of the present invention can be obtained as follows.

Flower pot ( Trichosanthes kirilowii maxim) is washed with water to remove debris, dried in the shade and crushed. Cranberries can be used without limitation, such as those grown or commercially available. Appropriate amount of solvent is added to the pulverized blend to ensure complete immersion. The extraction solvent is preferably a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, more preferably 80% ethanol. Next, the cheonhwabun extract obtained above is filtered and concentrated under reduced pressure to obtain a final extract.

The term " multidrug resistance " as used herein refers to a property in which a cell, particularly a particular disease-causing cell, is resistant to various drugs or compounds having very different chemical structures. According to a preferred embodiment of the present invention, the multiple drug resistance is multiple drug resistance by P-glycoprotein.

As used herein, the term "P-glycoprotein" means a kind of transporter protein present in the cell membrane having an ATP-binding cassette (ABC) - Glycoprotein binds to drugs and releases drugs out of the cells in an active transport mechanism using ATP as an energy source to confer drug resistance to the cells. In particular, P-glycoprotein expressed in cancer cells is known to increase the resistance to anticancer drugs, thereby significantly reducing the efficacy of chemotherapy.

Drugs that exhibit multiple drug resistance of the present invention include, for example, vinca alkaloids vinblastine, vincristine and navelbine; Taxanes family paclitaxel (TAX), taxotere; Doxorubicin, daunorubicin, epirubicin and idarubicin of the anthracyclines family; Epopodophyllotoxins based drugs etoposide and teniposide; Other drugs such as colchicine, mitoxantrone, dactinomycin, topotecan, trimetrexate, mithramycin, mitomycin C, But are not limited thereto.

The multidrug resistance-inhibiting effect of the composition of the present invention is characterized by its effect of inhibiting multiple drug resistance in cancer cells.

The cancer in the cancer cell can be, for example, an oral cancer, a stomach cancer, a lung cancer, a breast cancer, an ovarian cancer, a liver cancer, bronchogenic cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colon cancer, uterine cervical cancer, brain cancer, But are not limited to, prostate cancer, bone cancer, skin cancer, thymus cancer, parathyroid cancer, renal cancer or ureteral cancer.

The effective amount of the multi-drug resistant composition of the present invention and the anticancer agent-assisted composition is characterized in that it comprises 0.001 to 50% by weight of the active ingredient, which is the cheonhwa powder extract, based on the total weight of the composition of the present invention. In addition, the composition for suppressing multiple drug resistance and the composition for assisting an anticancer agent according to the present invention may further contain one or more active ingredients which exhibit the same or similar functions in addition to the above-mentioned effective ingredients. It may also be used alone or in combination with other chemotherapeutically active compounds, for example those well known in the art, such as vinblastine, adriamycin, cyclophosphamide, 5-FU, can be used as an anticancer adjuvant to inhibit the growth of cancer cells having multiple drug resistance to existing anticancer drugs in a suitable combination as well as binding with existing anticancer drugs such as amsacrine, daunomycin, and taxol.

The concentration of the active ingredient compound contained in the multi-drug resistance-inhibiting composition and the anticancer agent-assisting composition of the present invention can be determined in consideration of the purpose of treatment, the condition of the patient, the period required, and the like.

 The multi-drug resistance-inhibiting composition and the anticancer drug-adjuvant composition of the present invention may further comprise at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredient for administration. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The appropriate dose of the composition for inhibiting multiple drug resistance and the composition for adjuvant to an anti-cancer agent of the present invention may be appropriately determined depending on the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, It can be prescribed variously by the same factors. On the other hand, the oral dosage of the pharmaceutical composition of the present invention is preferably 0.001-100 mg / kg (body weight) per day.

The composition for inhibiting multiple drug resistance and the composition for adjuvant to an anti-cancer agent of the present invention can be administered orally or parenterally, and when administered parenterally, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, have. It is preferable that the route of administration of the pharmaceutical composition of the present invention is determined according to the type of the disease to be applied.

The composition for inhibiting multiple drug resistance and the composition for assisting an anticancer drug according to the present invention may be prepared by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those skilled in the art. Or by formulating it into a unit dose form or by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

In addition, the composition for suppressing multiple drug resistance and the composition for assisting an anticancer drug of the present invention can be provided as a food composition together with a pharmaceutically acceptable carrier.

The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, ice creams, alcoholic beverages, vitamin complexes, health supplements, and the like. .

The amount of natural extracts in the food or beverage may be added at 0.01 to 15% by weight of the total food weight, the health functional beverage composition may be added in a ratio of 0.02 to 10g, preferably 0.3 to 1g based on 100ml.

The food composition of the present invention may contain, as an additional ingredient, various flavors or natural carbohydrates, and the like, in addition to containing a natural extract of an active ingredient. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The aforementioned flavoring agents can advantageously be used natural flavoring agents (tautin), stevia extracts (for example rebaudioside A, glycyrzin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).

In addition to the natural extracts, the food composition may contain various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain natural fruit juice and fruit flesh for the production of fruit juice beverages and vegetable beverages.

The active ingredient cheonhwamin extract of the present invention is a natural substance with little toxicity and side effects, so can be used with confidence even during long-term use.

Hereinafter, the present invention will be described in detail by way of examples. The following examples are only intended to illustrate the invention and the invention is not limited by the following examples.

Experimental Example  1. Preparation of herb extracts

After quantification of 100 g of hwahwahwa 80% ethanol was added by using ultrasonic vibration twice for 30 minutes. The extract was filtered under reduced pressure using a 185 mm diameter paper filter (Whatman, Maidstone, England). After concentrating using a vacuum condenser (Eyela, Japan), the powder obtained by lyophilizing with a freeze dryer (Freezedryer, Matsushita, Japan) was dissolved in DMSO, vortexed and centrifuged to remove the insoluble material. Respectively.

Experimental Example  2. KB Of VB20  In the cell line MDR1  Expression analysis

The human epithelial cell line KB used for the experiment was purchased through the Korean Cell Line Bank (Korea), and the KB cell line (KB / VB20) resistant to vinblastine 20nM was distributed by Professor Sung-Kwan Lee. . (KB) and KB / VB20 cell lines were cultured in DMEM (Wellgene) medium supplemented with 5% (v / v) serum (heat-inactivated fetal bovine serum (Wellgene) and antibiotic-antimycotic (Gibco BRL) And cultured in a humidified incubator at 37 ° C containing 5% CO ₂ .

RT-PCR was performed using KB cell line, KB / KV20 cell line, A549 cell line, and Caco-2 cell line to confirm the expression of MDR1 in KB / VB20 cells used in this experiment. As a result, MDR1 was not expressed in the KB cell line as shown in Fig. It was confirmed that MDR1 is overexpressed in the KB / VB20 cell line. Therefore, the KB / VB20 cell line was used as a multiple drug resistant cell line in the following experiment .

Experimental Example  3. Rhodamine  123 assay Screening through

Rhodamine 123 is a fluorescent substance that can move between the inside and the outside of the cell by P-gp. By measuring the accumulation of this drug in the cells using a flow cytometer (FACS Calibur, Becton Dickinson), the degree of expression of P-gp is inferred It becomes an indicator. That is, when the accumulation level of this substance in the cell is high, the amount of P-gp expression is small, so that the amount of P-gp expression increases, and when the accumulation degree is low, the amount of P-gp expression is large. Both KB and KB / VB20 cell lines were prepared to be 1x10 ⁶ and 1.5x10 ⁶ cells / well in a 6-well plate, followed by pretreatment of the medicament for 1 hour followed by addition of rhodamine 123 (2.5 μM) alone, Dipipine (Nicardipine, NCD, 12.5 uM) was added at the same time and incubated for an additional hour. Next, the cells were collected using a scraper, washed twice with cold PBS, and suspended in 300 ml of PBS for flow cytometry. The degree of suppression of P-gp function by pharmacologic extracts was determined by comparison using the following equation.

% of control activity = Fluorescence sample / Fluorescence control x 100

(Here, the fluorescence sample indicates the degree of fluorescence measured after treatment with natural extract or drug, and the fluorescence control indicates the degree of fluorescence measured after treatment with rhodamine 123 alone.)

Rhodamine 123 assay was performed after treatment with 25, 50, and 100 ㎍ / mL concentrations of cheonhwamin in KB and KB / VB20 cell lines, respectively. As a result, there was no difference according to the concentration in the KB cell line, it was observed that the ability to discharge the Rhodamine 123 material out of the cell in accordance with the concentration in the extract of cheonhwa minutes in KB / KV20 cell line (Fig. 2). As a result, it was confirmed that the extracts of cheonhwamin have a functional inhibitory activity of P-gp.

Experimental Example  4. P- glycoprotein and MDR1  Gene expression analysis

In order to confirm that the ability of Rhodamine 123 substance to be released from the cell is controlled at the protein level, 50 μg / ml of medicinal material is treated for 2 hours, and then a sample is obtained and Western blotting is carried out to obtain P-glycoprotein The amount was confirmed. As a result, as shown in Fig. 3, the P-glycoprotein protein was not expressed in the KB cell line but only in the KB / VB20 cell line (Fig. 3A). In addition, the KB / VB20 cell line was observed to inhibit P-glycoprotein in the group treated with nicardipine (Nicardipine, NCD) known as a control P-glycoprotein inhibitor and a natural extract (FIG. 3B).

In addition, RT-PCR was performed with mRNA obtained by treating the cell line with a 50 μg / ml concentration of the extract for 2 hours to determine whether the extract was affected by the MDR1 gene expression step encoding P-glycoprotein. As shown in FIG. 4, MDR1 gene expression was not expressed in KB cell line but expressed only in KB / VB20 cell line (FIG. 4A). In addition, MCD gene was used in the positive control group of nicardipine and natural extract of KB / VB20 cell line. It was confirmed that expression was suppressed (FIG. 4B).

Experimental Example  5. YB -1 protein expression analysis

Y-box-binding protein-1 (YB-1) is a protein that binds to conserved nucleic acid, regulates transcription and translation, and increases the repair system when DNA is damaged. It is also known to modulate the expression of P-gp by modulating the MDR1 gene known as one of the target genes of YB-1.

(YB-1) protein, which is known to modulate the expression of P-glycoprotein, by western blot. As shown in Figure 5, it was confirmed that the expression of YB-1 protein in the positive control group of nicardipine and cheonhwamin extract treatment group.

Experimental Example  6. vinblastine Sensitivity change analysis

Cell proliferation was measured by MTT assay to determine whether the extract of cheonhwa increase the sensitivity to vinblastine, one of anticancer drugs, in KB cell line and KB / VB20 cell line. MTT assay is a MTS mixture ((3- (4,5-dimethylthiazol- 2-yl) -5, 3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2 H -tetrazolium, inner salt) and an electron coupling reagent ( This is an experiment to measure the number of viable cells by measuring the concentration of water soluble formazan produced by reducing an enzyme called succinic dehydrogenase in the mitochondria of living cells of phenazine methosulfate (PMS)).

After the cheonhwamin extract was pre-treated with KB / VB20 cell line for 2 hours, the vinblastine (vinblastine) treatment by concentration (0 ~ 100 nM) for 48 hours to measure the degree of cell proliferation. As shown in FIG. 6, it was shown that the natural extract of Nicardipine is effective in increasing its sensitivity at low concentrations of vinblastine. It was confirmed that the cheonhwamin extract can be used as a composition for inhibiting multi-drug resistance by increasing the sensitivity of anti-cancer agent in multi-drug resistant cells.

Hereinafter, examples of the pharmaceutical composition and the food composition of the present invention will be described. However, the present invention is not intended to be limited thereto but is specifically described.

Formulation example  1. Preparation of pharmaceutical compositions

1-1. Sanje  Produce

Cheonhwahwa Extract 2g

1g lactose

The above components were mixed and packed in airtight bags to prepare powders.

1-2. Manufacture of tablets

Cinnamon Extract 100mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

1-3. Preparation of capsules

Cinnamon Extract 100mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

1-4. Injection preparation

Natural Flower Extract 100 mg

Mannitol 180 mg,

Na2HPO412H2O 26 mg

Distilled water 2974 mg

After mixing the above components, an injection was prepared according to the usual production method.

Formulation example  2. Preparation of food composition

2-1. Preparation of health functional food

Natural Flower Extract 100 mg

Vitamin mixture quantity

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium 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 composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2-2. Health drink  Produce

Natural Flower Extract 100 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (9)

Pharmaceutical composition for inhibiting multidrug resistance to anticancer agents comprising ethanol extract of cheonhwahwa as an active ingredient. delete The composition of claim 1, wherein the natural ethanol extract is included in an amount of 0.001 to 50% by weight based on the total weight of the composition. 2. The composition of claim 1, wherein the multidrug resistance is multiple drug resistance by P-glycoprotein. 2. The composition of claim 1, wherein said multidrug resistance-inhibiting is multidrug resistance-inhibiting in cancer cells. Anticancer adjuvant pharmaceutical composition for enhancing anticancer activity comprising cheonhwabun ethanol extract as an active ingredient. 7. The composition of claim 6, wherein the ethanol extract of Chunhwa is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition. Anticancer adjuvant food composition for enhancing the anticancer activity comprising cheonhwabun ethanol extract as an active ingredient. 9. The composition of claim 8, wherein the ethanol extract of Chunhwa is comprised in an amount of 0.001 to 50% by weight based on the total weight of the composition.

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