KR102262317B1 - Composition for anticancer or cancer supplement comprising root extract of Rhododendron mucronulatum - Google Patents

Abstract

The present invention relates to an anticancer or anticancer auxiliary composition comprising a Rhododendron mucronulatum root extract, and the composition according to the present invention comprising the Rhododendron mucronulatum root extract as an active ingredient is suitable for breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, It has the effect of inhibiting the growth of cancer cells such as lung cancer, colorectal cancer and blood cancer or inducing apoptosis. In addition, the azalea root extract of the present invention is a natural substance, has no cytotoxicity, and is harmless to the human body, so it can be safely used in pharmaceutical and food compositions.

Description

Composition for anticancer or cancer supplement comprising root extract of Rhododendron mucronulatum

The present invention relates to a composition for adjuvant anticancer or anticancer comprising the root extract of Rhododendron mucronulatum , and more particularly, to a pharmaceutical composition for anticancer or a food composition for adjuvant anticancer comprising the root extract of Rhododendron as an active ingredient. .

Despite advances in modern medicine, cancer is still one of the most difficult human diseases to treat. Cancer is a tissue composed of abnormal cells that arise from one or more tissues and spread to other tissues. Currently, more than 100 known types of cancer are known ("Defining Cancer". National Cancer Institute. 17 September 2007. Retrieved 28 March 2018). It is known that 90-95% of the main causes of cancer are genetic mutations caused by environmental factors. Most of these factors are smoking, obesity, radiation, UV rays, stress, and environmental pollution (Anand P, Kunnumakkara AB, Sundaram). C, Harikumar KB, Tharakan ST, Lai OS, Sung B, Aggarwal BB (September 2008). "Cancer is a preventable disease that requires major lifestyle changes". Pharmaceutical Research. 25 (9): 2097-116).

Current cancer treatment methods include chemotherapy using drugs such as antimetabolites, alkylating agents, topoisomerase inhibitors, monoclonal antibodies, or radiation therapy at the same time. also use it

However, the use of these drugs may vary from patient to patient, but there are generally side effects such as hair loss, suppression of bone marrow function, drug resistance, and organ damage (Rachel Airley (2009). Cancer chemotherapy. Wiley-Blackwell. ISBN 978-0-470-09254-5). Numerous efforts are being made all over the world to conquer cancer, and as one of these efforts, research to prevent the proliferation of cancer using natural products is active.

On the other hand, rhododendron ( Rhododendron mucronulatum ) is a deciduous shrub of the dicotyledonous plant Rhododendron family Rhododendron, also referred to as a true flower or a rhododendron. It is distributed limitedly only in Korea, Japan, and China around the world, and grows in groups in sunny places. The flowers of azalea are collected in March-April and the roots are collected in September-October, and dried well in sunlight to be used as herbal medicine. Among them, the bark of azalea root contains about 7% tannin, and is known to have effects of rejuvenating blood, promoting blood circulation, hemostasis, and stopping pain. In oriental medicine, it has been used since ancient times to treat hematemesis, nosebleeds, menstrual irregularities, uterine bleeding, rectal ulcer bleeding, dysentery, rheumatism, and bruises.

As a document on the efficacy of azalea root, Korean Patent Registration No. 10-0829722 discloses the skin inflammation alleviation effect of azalea root extract as a cosmetic composition for anti-aging skin containing azalea root extract as an active ingredient. In addition, Korean Patent No. 10-1071785 has reported a composition for treating atopic dermatitis comprising taxifolin 3-O-β-D-glucopyranoside isolated from azalea root extract as an active ingredient.

However, there have been no reports on the anticancer effect of azalea root so far.

Accordingly, the present inventors continued their research on azalea root extract, and as a result, it was found that azalea root extract has an inhibitory effect on various cancers such as breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, lung cancer, colorectal cancer and blood cancer. The present invention was completed by finding the facts.

Accordingly, the technical problem to be solved in the present invention is to provide a material having excellent human stability while having anticancer activity against breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, lung cancer, colorectal cancer and blood cancer.

In order to solve the above technical problem, the present invention provides a pharmaceutical composition for anticancer comprising Rhododendron mucronulatum root extract as an active ingredient.

In addition, the present invention provides an anti-cancer supplementary food composition comprising the root extract of Rhododendron mucronulatum as an active ingredient.

The cancer may be one or more cancers selected from the group consisting of breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, lung cancer, colorectal cancer, and blood cancer.

The composition comprising the Rhododendron mucronulatum root extract of the present invention inhibits cell growth in cancer cells and induces apoptosis, thereby preventing breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, lung cancer, colon cancer and blood cancer. has anticancer activity against

As used herein, the term "extract" refers to a preparation obtained by squeezing rhododendron roots with an appropriate leachate and evaporating the leachate, and is not limited thereto, but is not limited thereto, but is an extract obtained by extraction, a diluted or concentrated solution of the extract, or an extract. It may be a dried product obtained by drying, a crude product or a purified product thereof.

In the present invention, the azalea root extract can be obtained using an extraction method and an extraction solvent known in the art, preferably water, anhydrous or hydrous lower alcohols having 1-4 carbon atoms (methanol, ethanol, propanol, butanol, etc.) ), a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, chloroform, or 1,3-butylene glycol as an extraction solvent.

In addition, the rhododendron root extract can be obtained through a conventional purification process in addition to the extraction method using the extraction solvent. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc. Through various purification methods additionally performed Azalea root extract can also be obtained through the obtained fraction.

According to one specific embodiment of the present invention, after grinding the rhododendron root in the shade, 0.5 to 20 times, preferably 1 to 15 times the weight of the dried sample of C1 to C4 such as water, ethanol, methanol, etc. Using a mixed solvent in which a lower alcohol or water and ethanol are mixed at a mixing ratio (kg/L) of 1:0.1 to 1:10, preferably 1:0.2 to 1:9, at room temperature for about 0.5 to 48 hours, preferably Azalea root extract is obtained by using an extraction method such as stirring extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction for 1 to 30 hours, preferably ultrasonic extraction, and then filtering, concentrating or drying the obtained extract under reduced pressure. can be obtained.

In the present invention, the azalea root extract includes all extracts, fractions and purified products obtained in each step of extraction, fractionation or purification, and dilutions, concentrates or dried products thereof.

According to one embodiment of the present invention, the azalea root extract inhibits cell growth in cancer cells and induces apoptosis, so that in the group consisting of breast cancer, ovarian cancer, lung cancer, kidney cancer, liver cancer, colorectal cancer and blood cancer It has a preventive or therapeutic effect on one or more selected cancers.

The composition of the present invention may contain rhododendron root extract in an amount of 0.0001 to 10% by weight, preferably 0.001 to 5% by weight, based on the total weight of the composition. When the content is less than 0.0001% by weight, the effect of inhibiting cancer cell growth and inducing apoptosis is too weak, and when it exceeds 10% by weight, the increase in the effect according to the increase of the content is insufficient, and the stability of the formulation is not secured. have.

According to one embodiment of the present invention, there is provided an anticancer pharmaceutical composition comprising an azalea root extract as an active ingredient.

The pharmaceutical composition according to the present invention includes a pharmaceutically acceptable carrier in addition to the azalea root extract. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. it's not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered to mammals such as rats, mice, livestock, and humans by various routes such as oral or parenteral, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or cerebrovascular It can be administered by intravenous injection.

The pharmaceutical composition is administered in a medically effective amount. In the present invention, a pharmaceutically effective amount means an amount sufficient to treat or prevent a disease at a reasonable benefit/risk ratio applicable to medical treatment or prevention, and the effective dose level is determined by the type and severity of the disease, the activity of the drug, the patient age, weight, health and sex of the patient, the patient's sensitivity to the drug, the time of administration of the specific extract used, the route of administration and the rate of excretion, the duration of treatment, factors including drugs used in combination with or concurrently with the specific extract used, and other medicines It may be determined according to factors well known in the art. In general, a dose of 0.1 to 1,000 mg/kg, preferably 10 to 100 mg/kg, based on an adult, may be administered once to several times a day. In addition, in the case of an external preparation, it is recommended to apply it once to 5 times a day in an amount of 1.0 to 3.0 ml based on an adult and continue it for at least 1 month. However, the scope of the present invention is not limited by the above dosage.

The pharmaceutical composition may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. It may be prepared by introduction into a dose container. At this time, the formulation may be formulated as a conventional formulation in the pharmaceutical field according to the therapeutic purpose, for example, tablets, capsules, powders, granules, suspensions, emulsions, syrups, emulsions, warning agents, ointments, sprays, oils It may be formulated as a preparation, gel, alcohol, tincture, bath, liniment, lotion, patch, pad, cream, and the like. In addition, it can be preferably used as an external preparation for the skin for the purpose of topical administration applied directly to the affected area, in this case, the form of an ointment, lotion, spray, gel, etc. is preferable. These external preparations for skin may also be included in a support base or matrix that can be directly applied to the treatment site, and examples of the support base include gauze or a bandage.

According to one embodiment of the present invention, there is provided a food composition for an anticancer supplement comprising an azalea root extract as an active ingredient.

The food composition according to the present invention may further include not only the azalea root extract as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents.

Examples of the carbohydrate include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

For example, when the food composition of the present invention is prepared as a drink, in addition to the azalea root extract of the present invention, a pear concentrate, a plant mixed extract, for example, Astragalus, cinnamon, Angelica, licorice, baekchul, sagebrush, Reishi mushroom extract, polydextrose, vitamins C, nicotinic acid amide, trisodium citrate, citric anhydride, and the like may be further included.

The azalea root extract may be usefully added to new functional medicines, beverages, health foods or health supplements.

On the other hand, since the azalea root extract of the present invention is a natural material, it is harmless to the human body, and has almost no toxicity and side effects, so it can be safely used even for long-term use, and can be safely applied to pharmaceutical and food compositions as described above. .

As such, the composition comprising the azalea root extract according to the present invention has an effect of inhibiting the growth of cancer cells such as breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, lung cancer, colorectal cancer and blood cancer, or inducing apoptosis. In addition, the azalea root extract of the present invention is a natural substance, has no cytotoxicity, and is harmless to the human body, so it can be safely used in pharmaceutical and food compositions.

1 shows the growth inhibitory activity of azalea root extract against breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, lung cancer, colorectal cancer and blood cancer.

Hereinafter, examples and the like will be described in detail to help the understanding of the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Example 1> Preparation of azalea root extract

After grinding and drying the rhododendron roots in the shade, the rhododendron roots were washed and dried as an azalea root extract using water 10 times the weight of the dried sample, and then extracted using water at 100° C. for 2 hours. The extract was filtered through a 5 μm filter, concentrated with a vacuum concentrator, and then frozen and dried to prepare a powder for extracting rhododendron roots.

<Example 2> Evaluation of cancer cell growth inhibitory efficacy of azalea root extract ( in vitro )

To evaluate the cancer cell proliferation inhibitory efficacy of the azalea root extract prepared in Example 1, 12 types of cancer cells (breast cancer: HCC38, HCC1143, MDA-MB-231, MDA-MB-453, AU565; kidney cancer: ACHN; brain tumor) : U-87 MG; Liver cancer: HepG2; Cervical cancer: HeLa; Lung cancer: A549; Colorectal cancer: HCT116; Hematological cancer: Jurkat) and breast tissue-derived epithelial cells (MCF10A) were evaluated using the MTT method. For this, 3-5 x 10 ³ cells per cancer cell were placed in each well of a 96-well plate and attached. After 24 hours, azalea root extracts were treated by concentration and cultured for 3 days, followed by addition of MTT reagent and incubated for 4 hours. Thereafter, the medium was carefully removed and the absorbance was measured at 570 nm by adding DMSO, and the results are shown in FIG. 1 . The survival rate of each well was calculated according to Equation 1 below.

As shown in FIG. 1, 12 types of cancer cells except for breast tissue-derived epithelial cells (MCF10A) (breast cancer: HCC38, HCC1143, MDA-MB-231, MDA-MB-453, AU565, kidney cancer: ACHN, brain tumor: U- 87 MG, liver cancer: HepG2 cervical cancer: HeLa, lung cancer: A549, colorectal cancer: HCT116, blood cancer: Jurkat), it was confirmed that azalea root extract had a concentration-dependent effect on cell proliferation inhibition.

<Example 3> Evaluation of apoptosis 0 inducing efficacy of azalea root extract

In order to evaluate the apoptosis-inducing ability of the azalea root extract prepared in Example 1, double staining with Annexin V/Propidium iodide was confirmed through flow cytometry. First, 12 cancer cells (breast cancer: HCC38, HCC1143, MDA-MB-231, MDA-MB-453, AU565; kidney cancer: ACHN; brain tumor: U-87 MG; liver cancer: HepG2; cervical cancer: HeLa; lung cancer: A549; colorectal cancer: HCT116, hematological cancer: Jurkat) was inoculated in a 6-well microplate containing cell culture solution ^{at a number of 4x10 5} cells/well, and incubated at 37° C. in a _{5% CO 2 incubator for 24 hours.} After that, each well was incubated for 4 hours with serum-free medium, replaced with a cell culture medium, treated with azalea root extract at a concentration of 10 and 50 μg/mL for 72 hours, and then stained with Annexin V and Propidium iodide. did. The stained cells were analyzed by flow cytometry (BD FACSCalibur™) and the results are shown in Table 1.

Live cell %
AnnexinV-/PI- Early
apoptosis%
AnnexinV+/PI- Late
apoptosis%
AnnexinV+/PI+ HCC38 0 μg/ml 85.0 5.2 9.8 10 μg/ml 32.5 12.7 54.8 50 μg/ml 11.6 20.6 67.8 HCC1143 0 μg/ml 83.0 6.9 10.1 10 μg/ml 47.5 13.2 39.3 50 μg/ml 14.1 21.3 64.6 MDA-MB-231 0 μg/ml 86.9 3.5 9.6 10 μg/ml 37 17.5 45.5 50 μg/ml 22.8 16.5 60.7 MDA-MB-453 0 μg/ml 83.1 5.5 11.4 10 μg/ml 76.1 9.5 14.4 50 μg/ml 27.5 14.7 57.8 AU565 0 μg/ml 80 9.3 10.7 10 μg/ml 46.8 12.0 41.2 50 μg/ml 10.7 18.9 70.4 ACHN 0 μg/ml 82.1 8.7 9.2 10 μg/ml 73.5 11.4 15.1 50 μg/ml 18.9 19 62.1 U-87 MG
0 μg/ml 81.6 7.9 10.5 10 μg/ml 69.6 12.8 17.6 50 μg/ml 22.5 17.2 60.3 HepG2 0 μg/ml 82.5 6.7 10.8 10 μg/ml 82.5 6.7 10.8 50 μg/ml 21.4 13.8 64.4 HeLa 0 μg/ml 84.7 6.4 8.9 10 μg/ml 71.6 12.7 15.7 50 μg/ml 30.8 20.3 48.9 A549 0 μg/ml 82.9 7.1 10.0 10 μg/ml 74.3 10.7 15 50 μg/ml 28.7 19.8 51.5 HCT116 0 μg/ml 83.1 5.8 11.1 10 μg/ml 80 7.2 12.8 50 μg/ml 39.5 11.2 49.3 Jurkat 0 μg/ml 80.5 6.3 13.2 10 μg/ml 55.5 12.7 31.8 50 μg/ml 29.5 13.6 56.9

As shown in Table 1 above, cells stained with both AnnexinV/PI, which means apoptosis (programmed cell death) by azalea root extract, were found in 12 types of cancer cells (breast cancer: HCC38, HCC1143, MDA-MB-231, MDA-MB- 453, AU565, kidney cancer: ACHN, brain tumor: U-87 MG, liver cancer: HepG2 cervical cancer: HeLa, lung cancer: A549, colorectal cancer: HCT116, blood cancer: Jurkat) in a concentration-dependent manner.

Formulation Example 1: Pharmaceutical Formulation

1-1. Preparation of powders

ingredient Weight (g) Rhododendron Root Extract 2 lactose One

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

1-2. manufacture of tablets

ingredient Weight (mg) Rhododendron Root Extract 100 corn starch 100 lactose 100 magnesium stearate 2

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

1-3. manufacture of capsules

ingredient Weight (mg) Rhododendron Root Extract 100 corn starch 100 lactose 100 magnesium stearate 2

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

Claims (5)

A pharmaceutical composition for anticancer comprising Rhododendron mucronulatum root extract as an active ingredient. The method of claim 1,
The cancer is breast cancer, kidney cancer, brain tumor, liver cancer, cervical cancer, lung cancer, a pharmaceutical composition for anticancer, characterized in that at least one cancer selected from the group consisting of colorectal cancer and blood cancer. The method of claim 1,
The composition inhibits cancer cell proliferation and a pharmaceutical composition for anticancer, characterized in that inducing apoptosis in cells. The method of claim 1,
The anticancer pharmaceutical composition, characterized in that the azalea root extract is included in an amount of 0.0001 to 10% by weight based on the total weight of the composition. Rhododendron mucronulatum ) An anticancer supplementary food composition comprising a root extract as an active ingredient.

Images