WO2014077435A1 - Composition containing inulae radix extract for renal cancer treatment cosmetics - Google Patents

Abstract

The present invention relates to a novel use of a Inulae Radix extract, in which a composition for treatment and cosmetics contains, as an active ingredient, a Inulae Radix extract exhibiting excellent preventative and treatment effects on renal cancer. The Inulae Radix extract according to the present invention suppresses growth and induces apoptosis of renal cancer cells, and therefore can be used effectively for renal cancer treatment and prevention.

Description

Composition and cosmetic composition for treatment of kidney cancer comprising civil extract

The present invention relates to a novel use of civil extract. Specifically, the present invention relates to a therapeutic composition and cosmetic composition containing a civil extract as an active ingredient exhibiting excellent prophylactic or therapeutic efficacy against kidney cancer.

Among the major diseases of modern people, researches on the treatment and diagnosis of cancer are being conducted mainly on lung cancer, liver cancer, and stomach cancer, which have high incidence.

The kidney is an important urinary system that has the role of excreting waste products in vitro by filtering blood to produce urine. At the same time, it is also an important endocrine organ that produces hormones such as angiotensin, which regulates blood pressure, and erythropoietin, a red blood cell hematopoietic factor. Renal tumors include kidney cell cancer in adults, Wilms' tumor in children, and sarcoma as a rare tumor, but the most common malignancy below Renal cell cancer, a tumor, is called kidney cancer. The incidence of renal cancer is about 2.5 per 100,000 population, and male to female ratio is 2 to 3: 1. Among urological malignancies, there are many tumors following prostate cancer and bladder cancer.

Genetic factors are also known as risk factors for kidney cancer, but generally include smoking and excessive fat intake. It is also known that the incidence of such tumors is high in patients undergoing long-term dialysis. In kidney cancer, when the tumor has a maximum diameter of 5 cm or less, some subjective symptoms rarely occur and are often found by CT scan or the like at the time of examination. Larger tumors may include hematuria, abdominal tumors, and pain. In addition, systemic symptoms may cause fever, weight loss, anemia and the like, and rarely, endocrine factors may cause erythrocytosis, hypertension, hypercalcemia, and the like. On the other hand, there may be cases in which vein veins of the abdominal body surface or venous varicose veins occur due to the progression of kidney cancer into the inferior vena cava. About 20% of kidney cancers are found from lung or bone metastases. In kidney cancer, tumors tend to spread in veins, and metastasis to other organs is likely to occur.

Ultrasound, CT, angiography, etc. are examined for kidney cancer, but specific biochemical markers are not known. Therefore, examination using a device is necessary. In addition, although several types of kidney cancer can be classified pathologically, the cause of the development of clear cell type kidney cancer, which accounts for about 90%, is known to be a defect in the von-Hippel-Lindau (VHL) gene, a cancer suppressor gene. . Deletion of the VHL gene results in transcriptional activation of the hypoxic inducible gene group through association of HIF-α / VHF, leading to enhanced expression of genes such as VEGF and TGFβ.

The main treatment for kidney cancer is surgical therapy. Regardless of the timing of the disease, extraction of the kidneys or partial extraction of the kidneys is the most common where possible, and even if there is metastasis, surgical removal of the kidneys may be considered. Arterial embolization of the renal artery may be used as a method other than the surgical therapy. This method may be performed prior to surgery in cases where it is impossible to extract or when a large tumor is removed. Recent advances in burn diagnostics have led to early detection of renal cancer, even at very small sizes, resulting in more than 90% healing in early cancer. However, since the therapeutic results of large tumors larger than 5 cm or tumors with metastases are poor (the 5-year survival rate of all renal cell carcinomas is about 50 to 60%), the importance of early detection is recognized.

Inulae Radix ( Inulae Radix ; Inulae Radix L.), a perennial herb of the Asteraceae family, is a cultivated plant of European origin, with a straight stem and short hairs on its predecessor, 80 cm to 2 m high. In Korea, it refers to the root of the aroma of the Asteraceae. The leaves are broad oval or long oval, with sharp pointed ends, narrowed at the bottom, flowing into the foliar, with irregular sawtooth at the edges, and the leaves at the bottom are foliated but disappeared upward and wrap the main stem with the bottom. Flowers are yellow and hang on the top of stem in July-August. Guns are hemispherical, outer shells are ovate, leaf-like, and short hairs grow. Fruits are achene with light reddish brown tubules. Root contains inulin, essential oil, and alantopicrin in the leaves. Mok-hyang, unlike its name, is herbaceous and originally called scent of honey because it smells like honey. It also has five roots in one tree, five branches in one stem, five leaves in one branch, and five nodes in each branch. The fragrance spreads. This medicine has a distinctive scent, tastes spicy, bitter and warm.

]. It is used for abdominal pain, gastrointestinal disorder, vomiting, diarrhea, bronchitis, whooping cough, chronic enteritis, tuberculosis, and fetal and maternal fetus. Pharmacological action has been reported insecticide, antibacterial action, intestinal, uterine exercise excitement, peripheral paralysis action. The shape is columnar or conical and some have side roots. The thick one is split vertically, the outer side is brown, wrinkled and sloppy. When looking at the cross section with a magnifying glass, the skin is grayish brown and the neck is distinctly grayish white. The roots of the cedar in Chinese medicine and civilian are called the cedar or the incense, and it is used as a medicine for phlegm, earth and sand, abdominal pain, vomiting, bronchitis, whooping cough, chronic enteritis, tuberculosis, nursing hari, stomach cramps, and malaria. do.

However, nothing has been disclosed or disclosed in the above literature with regard to diseases related to civil cancer kidney cancer.

Accordingly, the present inventors completed the present invention by confirming that the extract of the civil fragrance can effectively kill kidney cancer cells while studying the herbal medicine for the civil fragrance.

It is an object of the present invention to provide a composition and a cosmetic composition for treating renal cancer containing a civil extract as an active ingredient.

In order to achieve the above object, the present invention provides a composition for preventing and treating kidney cancer comprising an active ingredient extracted with an organic solvent civil engineering fragrance. It is preferable that the said organic solvent is ethanol.

In another aspect, the present invention provides a cosmetic composition for preventing kidney cancer, comprising a civil ethanol extract containing cosmetically acceptable cosmetic supplement additives as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for preventing and treating kidney cancer, which contains a civil ethanol extract as an active ingredient. The composition of the present invention comprises a civil extract as an active ingredient, and may further comprise a pharmaceutically acceptable carrier or diluent.

In the composition for preventing and treating kidney cancer of the present invention, the ethanol extract is preferably extracted at 50 ° C. for 24 hours, wherein the ethanol extract is more preferably dried and concentrated at 45 ° C. under reduced pressure, and the ethanol Is most preferably 95%.

In addition, in the composition for preventing and treating kidney cancer of the present invention, the kidney cancer is preferably renal cell cancer.

The civil extract of the present invention is possible at any part of the civil fragrance, but is preferably extracted from the civil fruit. The extraction solution may be obtained by extraction with water or an organic solvent, and examples of the organic solvent may include lower alcohols, acetone, chloroform, methylene chloride, ether, ethyl acetate, and hexane. Lower alcohols include methanol, ethanol, propanol and butanol, with ethanol being most preferred.

Specifically, 1 to 5 times, preferably 3 times, 95% ethanol is added to civil engineering dry matter or powder, and 10 to 100 hours, preferably 15 at a temperature of 20 to 100 ° C, preferably 40 to 60 ° C. To 40 hours, more preferably, the extract for 24 hours and then filtered to produce a civil ethanol extract of the flavor. Preferably, the filtrate obtained by filtering the extract may be concentrated under reduced pressure. In the above extraction methods, the extraction process may be repeated two or more times as necessary, and the extract obtained after filtration may be lyophilized or dried under reduced pressure to obtain a powder form.

The "pharmaceutically acceptable carrier" is a pharmaceutically acceptable substance such as a liquid or solid filler, diluent, excipient or solvent which serves to transport the active ingredient from one organ or part of the body to another organ or part of the body. , Composition or vehicle.

The composition for treating kidney cancer of the present invention may be prepared as a medicament by adding one or more pharmaceutically acceptable carriers together with the active ingredient. The carrier may include, but is not limited to, saline, buffered saline, water, glycerol and ethanol, and any suitable agent known in the art (Remingtons's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA) may be used. .

Formulations for pharmaceutical formulation civil extract of the present invention can be administered orally at the time of clinical administration and can be used in the form of general pharmaceutical formulations, when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants And diluents such as surfactants or excipients. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and liquid preparations for oral use include suspensions, solvents, emulsions, and syrups. In addition to liquids and paraffins, various excipients may be included, such as wetting sweeteners, fragrances, preservatives and the like.

In addition, the herbal medicine that may be added to the composition of the present invention may be any pharmaceutically acceptable herbal medicine, for example, Angelica tenuissimae Radix, Gastrodiae Rhizoma, Bapleuri Radix, Angelica ( Angelicae gigantis Radix, Persicae Semen, Cinnamomi Ramulus, Rhubarb (Rhei Rhizoma), Licorice (Glycyrrhizae Radix), Cnidii Rhizoma, Aurantii nobilis Pericarpium, Taxa (Alismatis Rhizoma) Coptidis Rhizoma, Scutellariae Radix, Hoelen, Peeoniae Radix, Atractylodis Rhizoma alba, Phellodendri Cortex, Gardeniae Fructus, Pinelliae Tuber, Ramulu Set (Uncaria) Uncus, Ponciri Fructus, Ginseng, Gingseng, Liriopis Tuber, Polygalae Radix, Acori graminei Rhizoma, Atractylodis Rhizoma alba, Chrysanthemi Flos, Windproof Radix ), Ginger (Zingiberis Rhizoma crudus), forget-me-not (Natrii sulfas), control (Ziz) yphi Fructus, Salviae Radix, Mautan Radicis Cortex, Rehmanniae Radix, Mint Herba, Dioscoreae Rhizoma, Polyporus, Polygoni multiflori Radix, Gusi (Allii tube) Semen, Cassiae Semen, Lycii Fructus, Araliae cordatae Radix, Eucommiae Cortex, Hedyotis Herba, Saururus Herba, Artemisiaecapillaris Herba, Anemarrhen Rhizoma, Carthami Flos, Astragali Radix, Lycopodium, Ginkgonis Folium, Polygonati Rhizoma, Nelumbinis Semen, Fossilia ossis Mastodi, Cortexi radics ), Achyranthis Radix, Rehmanniae Radix preparata, Perillae Semen, Thujae Semen, Malt (Hordei Fructus germinatus), Cuscutae Semen, Mordaedae Radix, Sea Song (Pini koraiensis) Radix) and the like can be used alone or in combination.

The composition of the present invention may be administered in various parenteral formulations during actual clinical administration, and solid preparations include tablets, pills, powders, granules, capsules, and the like. In addition to water, liquid, and paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Specifically, preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used. In addition, calcium or vitamin D ₃ may be added to enhance the efficacy of the treatment. Such compositions may be presented in unit-dose (single) or multi-dose (several) containers, such as sealed ampoules and vials, and immediately before use, sterile liquid carriers such as injectable water. Can be stored under freeze-drying conditions requiring only the addition of. Immediate injection solutions and suspensions can be prepared from sterile powders, granules and tablets.

The formulations of the present invention can be applied differently depending on the age, sex, condition of the subject, the absorption of the active ingredient in the body, the inactivation rate and excretion rate, the drug used in combination. The invention also includes formulations of dosage units. The formulations are present in individual dosage forms, such as tablets, coated tablets, capsules, pills, suppositories, and ampoules, wherein the amount of active compound in the drug corresponds to the fraction or multiple of the individual dosage. Dosage units may contain, for example, one, two, three or four times the individual dosage, or 1/2, 1/3 or 1/4 times. The individual dosages preferably contain an amount in which the active compound is administered at one time, which usually corresponds to all, 1/2, 1/3 or 1/4 times the daily dosage.

As used herein, the term "extract" refers to an active ingredient isolated from natural products. The extract may be obtained by an extraction process using water, an organic solvent, or a mixed solvent thereof, and includes an extract, a dry powder thereof, or any form formulated using the same.

In a specific embodiment of the present invention, the civil ethanol extract killed 98.8% ACHN kidney cancer cells at 100 μg / ml, and IC ₅₀ (half maximal inhibitory concentration) was 10.3 μg / ml. The above results demonstrate that the civil extract of the present invention has excellent killing activity of ACHN kidney cancer cells and further has therapeutic and prophylactic activity for kidney cancer.

As used herein, the term "prevention" means any action that inhibits or delays the development of kidney cancer by administration of the composition.

As used herein, the term "treatment" means any action that improves or advantageously changes the symptoms of kidney cancer by administration of the composition.

Civil extract in the present invention can be used to extract using water, an organic solvent, or a mixed solvent thereof. Preferably it is extracted using an organic solvent, in particular ethanol. The extracted solution can be used directly or can be concentrated and / or dried. When extracted with an organic solvent, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof may be used and extracted by room temperature or warming under conditions where the active ingredient of the herbal medicine is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and loss of the active ingredient of the drug may vary, so select an appropriate organic solvent. The extraction method is not particularly limited, and examples thereof include cold needle extraction, ultrasonic extraction, reflux cooling extraction, and the like. Filtration is a process of removing the suspended solid particles from the extract, it may be used to filter the particles using cotton, nylon or the like, or may be used, such as ultrafiltration, cryofiltration, centrifugal separation, but is not limited thereto.

Concentration of the extract may be used, such as concentrated under reduced pressure, reverse osmosis concentration. The drying step after concentration includes freeze drying, vacuum drying, hot air drying, spray drying, reduced pressure drying, foam drying, high frequency drying, infrared drying, and the like. If desired, a process of grinding the final dried extract may be added.

In addition, the extract can perform an additional fractionation process. Preferably, the extract is suspended in distilled water to obtain a nonpolar solvent soluble layer by extraction and separation with a nonpolar organic solvent such as hexane, ether, dichloromethane, chloroform, ethyl acetate, or a mixed solvent thereof. It can be used by concentrating and / or drying it.

In the present invention, the term "pharmaceutically acceptable salts" means salts derived from pharmacologically or physiologically acceptable inorganic acids, organic acids and bases. Examples of suitable acid include hydrochloric acid, bromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, Formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium, alkaline earth metals such as magnesium, ammonium and the like.

The pharmaceutical composition for preventing and treating kidney cancer diseases of the present invention comprises 0.1 to 50% by weight of the extract or compound based on the total weight of the composition. In addition, the composition does not increase the efficacy, but may include additional ingredients that are commonly used in the pharmaceutical composition to improve the smell, taste, time and the like. In addition, the composition adds inorganic and organic additives such as vitamins B1, B2, B6, C, E, niacin, carnitine, betaine, folate pantothenic acid, biotin, zinc, iron, calcium, chromium, magnesium, and mixtures thereof. It can be included as. In addition, the composition may include a substance having a therapeutic activity against kidney cancer used alone or used in the past.

As used herein, the term "patient" refers to humans, horses, sheep, pigs, goats, and camels having a disease caused by kidney cancer and its direct and indirect causes, and whose symptoms may be improved by administering the composition of the present invention. Means animals such as nutrition, dogs. By administering to the patient a composition comprising a civil extract of the present invention, the above-mentioned kidney cancer can be effectively prevented and treated. The composition of the present invention can be administered in parallel with existing kidney cancer treatments.

As used herein, the term "administration" means introducing a predetermined substance into a patient by any suitable method, and the route of administration of the composition of the present invention is oral or parenteral via any general route as long as the target tissue can be reached. May be administered. In addition, the composition may be administered by any device in which the active agent may migrate to the target cell.

The composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level refers to a patient's sexually transmitted disease, age, severity, and drug activity. , Drug sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs, and other factors well known in the medical arts. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. It may be single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, and can be easily determined by those skilled in the art. The method of administering the composition comprising the extract or compound prepared according to the preparation method of the present invention is preferably oral administration or intravenous administration. In general, when the effective dose is oral administration, it is usually 1 to 1 time per adult. 500 mg / kg is preferred, and in the case of intravenous administration, 1 to 100 mg / kg is preferred, and may be administered 2-3 times a day. Dosage levels for a particular patient may vary depending on gender, age, civil orientation, diet, time of administration, method of administration, drug mixture, the condition of the patient, and the extent of neurological disease.

In addition, the present invention provides a cosmetic composition for preventing kidney cancer, which comprises a civil ethanol extract comprising an cosmetically acceptable cosmetic auxiliary additive as an active ingredient.

The cosmetic composition of the present invention preferably contains 0.01 to 10% by weight, and more preferably 0.1 to 1% by weight, based on the total weight of the ethanol extract. The cosmetic composition of the present invention is not particularly limited in the formulation, for example, it may have a flexible cosmetic water, nourishing cosmetics, massage cream, nutrition cream, pack, gel or skin adhesive type cosmetic formulation, and also lotion, It may be a transdermal dosage form such as an ointment, gel, cream, patch or spray.

In the present invention, when the civil extract of the present invention was injected into ACHN kidney cancer cells, it was confirmed that ACHN cells are killed (see FIG. 1 ). Therefore, it can be seen that civil extract is effective in treating kidney cancer. Thus, in the present invention, the present invention was completed by preparing a cosmetic composition for preventing kidney cancer, which contains civil extract as an active ingredient (see Preparation Example 2 ).

As discussed above, the civil extract of the present invention inhibits the growth of kidney cancer cells and induces apoptosis. Therefore, the composition for treating renal cancer according to the present invention will be very effective in the treatment of renal cancer patients.

1 is a result of the Alamar Blue analysis to determine the effect of the introduction of civil fragrance on the growth of renal cancer cells in ACHN cells, human kidney cancer cells, wherein X axis is the concentration of civil extract extract, Y axis is the surviving human kidney The survival rate of cancer cells is shown.

Figure 2 is the EdU assay results to determine whether the introduction of the civil fragrance obtained in Example 1 in human kidney cancer cell line ACHN cells to stop the cell cycle and inhibit proliferation of kidney cancer cells.

Figure 3 is a reporter assay to determine the effect of the introduction of civil fragrance obtained in Example 1 on the renal cancer cell c-Myc cell signal in ACHN cells, a human kidney cancer cell line, the activity of c-Myc cell signal The result of comparative analysis.

Figure 4 is a reporter assay to determine the effect of the introduction of the civil fragrance obtained in Example 1 on the AP-1 cell signal of kidney cancer cells in ACHN cells, a human kidney cancer cell line, the activity of AP-1 cell signal The result of comparative analysis.

Figure 5 is a reporter assay to determine the effect of the introduction of the civil fragrance obtained in Example 1 on the NF-κB cell signal of kidney cancer cells in ACHN cells, a human kidney cancer cell line, the activity of the NF-κB cell signal The result of comparative analysis.

Hereinafter, the present invention will be described in more detail based on the following examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited to the following examples and may be changed to other embodiments equivalent to substitutions and equivalents without departing from the technical spirit of the present invention. Will be apparent to those of ordinary skill in the art.

Example 1 Preparation of Civil Engineering Extract

3 kg of civil fragrance (Chinese) purchased from Seoul medicinal herb were dried and ground for 5 days at the shade and room temperature. The ground civilization was immersed in 30 L of 95% ethanol and extracted at 50 ° C. for 24 hours. This was filtered through filter paper, dried and concentrated under reduced pressure at 45 ° C. to obtain 512 g of the total extract, which was stored at −20 ° C.

Example 2 Effects of Civil Engineering Extracts on the Growth of Kidney Cancer Cells

In order to examine the effects of the civil extract extracted in Example 1 on the growth of kidney cancer cells, ACHN cells, which are human kidney cancer cells, were treated with civil ethanol extracts for 48 hours and subjected to Alamar Blue analysis. The Alamar Blue assay is a modified form of the MTT assay, in which a specific enzyme degrades a living cell and then measures the fluorescence intensity of the product as the compound breaks down to determine the relative number of living cells after treatment. I am an experimental method. It will be described in more detail below.

<2-1> Preparation and Treatment of Human Kidney Cancer Cell Line

ACHN cells, a kidney cancer cell line used in the present invention, were distributed from Korean Cell Line Bank (KCLB) and used for experiments. Specifically, ACHN kidney cancer cells (Minimum essential medium) containing 10% FBS (fetal bovine serum, fetal bovine serum) (Welgene) and 25 mM HEPES (4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid) Subcultured in medium.

<2-2> Cell growth inhibition measurement of ACHN kidney cancer cells

The civil extract extracted in Example 1 was confirmed the effect of inhibiting growth against ACHN cells which are kidney cancer cells. Specifically, 6.0 x 10 per well in a 96 well plate³ The civil fragrance dissolved in dimethyl sulfoxide (DMSO) after seeding and incubating for 24 hours in ACHN kidney cancer cells When the ethanol extract was treated at 0 to 100 μg / ml concentrations (specifically, 0, 3.125, 6.25, 12.5, 25, 50 and 100 μg / ml concentrations) for 48 hours, the degree of inhibition of cell growth was confirmed. (Table 1). After treatment with each concentration of extract, 20 μl of Alamar Blue reagent was added to 0.2 ml of cell culture filled in each well in a 96-well plate, and the plates were incubated for 2 hours in an incubator. The plate was slowly shaken to evenly react the cells in each well, and the intensity of fluorescence was measured at 590 nm with a Fluorescence Microplate Reader (Molecular Devices Corp.) while irradiating irradiation light at a wavelength of 544 nm. The survival rate of is shown in FIG.

Table 1

Civil flavor	Kidney Cancer Cell Survival (Average)	Standard Deviation
0 μg / ml	1.000	0.000
3.125 μg / ml	0.996	0.055
6.25 μg / ml	0.753	0.020
12.5 μg / ml	0.381	0.032
25 μg / ml	0.119	0.010
50 μg / ml	0.010	0.001
100 μg / ml	0.012	0.002

As a result, as shown in Table 1 and Figure 1, the higher the treatment concentration of civil fragrance, the growth of kidney cancer cells was reduced, from which it can be seen that civil engineering has a renal cancer treatment effect. Ie 0.4% at 3.125 μg / ml, 24.7% at 6.25 μg / ml, 61.9% at 12.5 μg / ml, 88.1% at 25 μg / ml, 99.0% at 50 μg / ml, and 98.8% at 100 μg / ml Kidney cancer cells were killed. In addition, the IC ₅₀ (half maximal inhibitory concentration) was determined to be 10.3 μg / ml. Table 1 describes the relative cell numbers of kidney cancer cells after 48 hours according to the concentration of each civil fragrance based on the number of survival rates of kidney cancer cells of the control group not treated with civil fragrance. As such, the civil extract of the present invention demonstrates that it has excellent ACHN cell killing activity and further has renal cancer treatment and prophylactic activity.

<Example 3> Acute toxicity test by civil extract

The civil fragrance used in the present invention was widely used as a medicinal herb, so it was judged that there would be no problem in stability.

Acute toxicity test was performed using 6-week-old SPF SD rats. Two animals per group were suspended orally administered at a dose of 5 g / kg, each of which was suspended in 0.5% methylcellulose solution of the civil extract of Example 1 of the present invention. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to observe abdominal and thoracic organ abnormalities.

As a result, there were no clinical symptoms or deaths in all animals treated with the test substance, and no toxicity change was observed in weight change, blood test, blood biochemistry test, autopsy findings, etc. As a result, all civil extracts did not show toxic changes up to 5 g / kg in rats and the minimum lethal dose (LD ₅₀ ) was determined to be a safe substance above 5 g / kg.

Example 4 Cell Proliferation Test (EDU Assay)

EdU assay is the same test method as BrdU assay and is an improved assay. Halogenated nucleotides such as the pyrimidine analog bromodeoxyuridine (BrdU) are useful for staining DNA in living cells or tissues. In phase S, which is doubled in the nucleus of the cell cycle and doubled in volume, BrdU is used instead of thymidine in the replicating DNA, and the amount of synthesized DNA can be confirmed by staining with BrdU antibody.

In the test method, civil cancer was treated by concentration to kidney cancer cells in culture, and after 6 hours, EdU was added to the medium and the cells were cultured. After 18 hours, the medium containing EdU was removed and the cells were fixed with 3.7,% paraformaldehyde and washed twice with PBS. After permeation of cells with 0.1% Triton X-100, Alexa Fluor 488 azide was reacted with EdU. Stained with Hoechst 33342 to identify the nuclei of the cells. As shown in Fig. 2, the nucleus of the whole cell is the first one, of which the nuclei of the newly generated cell after the point of civil engineering treatment are the second one. The third is the sum of these.

As shown in Figure 2, when the civil fragrance was treated for each concentration, it can be seen that the number of newly generated cells compared to the untreated control. This shows that civil fragrance has an effect of inhibiting the proliferation of kidney cancer cells.

Example 5 Cell Signaling Test (Reporter Assay)

A reporter gene assay was performed to investigate the effect of civil fragrance on the major cellular signals that control the growth and proliferation of cancer cells. Reporter gene assay is a test technique designed to confirm the change of the cell signal by introducing a reporter gene that can measure the change of a specific cell signal in the cell.

In the test method, a reporter gene designed to measure a cell signal to be confirmed was transfected into cancer cells, stabilized, and civil aroma was processed at each concentration. After 24 hours, the expression level of the reporter gene was compared with 100% in the group without civil engineering and 0% in the group without transduction.

As a result of confirming the activity of the cell signal, it was confirmed that all the major signals for promoting the proliferation of kidney cancer cells decreased when the civil extract of 100 ug / ml was treated.

<Preparation Example 1> Preparation of a renal cancer treatment agent containing a civil extract as an active ingredient

The present inventors have confirmed that the renal cancer therapeutic effect of civil engineering extract through the above example was excellent, and prepared a renal cancer therapeutic agent containing the extract as an active ingredient as follows. In addition, the preparation of the following therapeutic agents can be used for the application of not only therapeutic agents but also cosmetic compositions.

<1-1> Soft gelatin capsules containing civil extract (wt%)

Civil extract 20%, Vitamin C 4.5%, Vitamin D ₃ 0.001%, Manganese sulfate 0.1%, Beeswax 10%, Palm oil 25%, Safflower seed oil 30.399%

 <1-2> Preparation of Intravenous Formulation Containing Civil Engineering Extract (wt%)

Civil extract 0.2%, mannitol 0.3%, saline 9.5%

<1-3> tablet (wt%) containing civil extract

Civil extract 35%, Vitamin C 10%, Vitamin D ₃ 0.001%, Manganese sulfate 0.1%, Crystalline cellulose 25.0%, Lactose 17.999%, Magnesium stearate 2%

Preparation Example 2 Preparation of Cosmetic Composition Containing Civil Engineering Extract as an Active Ingredient

The inventors have confirmed that the civil extract is excellent in the renal cancer treatment activity through the above embodiment to prepare a cosmetic composition containing it as an active ingredient as follows.

<2-1> Nutrient cosmetics containing a civil extract (% by weight)

Nutrients were prepared in a conventional manner according to the composition described below.

Glycerin 8.0%, Butylene Glycol 4.0%, Hyaluronic Acid Extract 5.0%, Betaglucan 7.0%, Carbomer 0.1%, Civil Extract 0.05%, Capilic / Capric Triglyceride 8.0%, Squalane 5.0%, Cetearyl Glucoside 1.5%, sorbitan stearate 0.4%, cetearyl alcohol 1.0%, triethanol amine 0.1%, remaining amount of purified water

<2-2> Nutrition cream containing civil extract (% by weight)

The nourishing cream was prepared by a conventional method according to the composition described below.

Glycerin 3.0%, Butylene Glycol 3.0%, Liquid Paraffin 7.0%, Beta Glucan 7.0%, Carbomer 0.1%, Civil Extract 3.0%, Capric / Capric Triglyceride 3.0%, Squalane 5.0%, Cetearyl Glucoside 1.5 %, Sorbitan stearate 0.4%, polysorbate 60 1.2%, triethanol amine 0.1%, purified water balance

<2-3> Massage cream containing civil extract (% by weight)

Massage creams were prepared in a conventional manner according to the composition described below.

Glycerin 8.0%, Butylene Glycol 4.0%, Liquid Paraffin 45.0%, Beta Glucan 7.0%, Carbomer 0.1%, Civil Extract 1.0%, Capric / Capric Triglyceride 3.0%, Beeswax 4.0%, Cetearyl Glucoside 1.5 %, Sesqui oleic acid sorbitan 0.9%, petrolatum 3.0%, paraffin 1.5%, remaining amount of purified water

<2-4> Pack containing civil extract extract (% by weight)

Packs were prepared by conventional methods according to the compositions described below.

Glycerin 4.0%, polyvinyl alcohol 15.0%, hyaluronic acid extract 5.0%, betaglucan 7.0%, allantoin 0.1%, civil extract 0.5%, nonyl phenylether 0.4%, polysorbate 60 1.2%, ethanol 6.0%, remaining amount of purified water

<2-5> Skin external preparation ointment containing civil extract (wt%)

Ointments were prepared in a conventional manner according to the compositions described below.

Glycerin 8.0%, Butylene Glycol 4.0%, Liquid Paraffin 15.0%, Beta Glucan 7.0%, Carbomer 0.1%, Civil Extract 3.0%, Squalane 1.0%, Cetearyl Glucoside 1.5%, Sorbitan Stearate 0.4%, Three Tearyl alcohol 1.0%, beeswax 4.0%, purified water balance

On the other hand, the specific scope of the present invention is defined by the claims rather than the embodiments described above, all changes and modifications derived from the meaning and scope and equivalent concepts of the claims to the scope of the invention It should be interpreted as including.

Claims (7)

1. Composition for preventing and treating kidney cancer comprising an active ingredient extracted with an organic solvent civil engineering fragrance.
2. The method of claim 1, wherein the organic solvent is a composition for preventing and treating kidney cancer, characterized in that ethanol.
3. According to claim 2, wherein the active ingredient extracted with ethanol is a composition for preventing and treating kidney cancer, characterized in that extracted for 24 hours at 50 ℃.
4. According to claim 3, wherein the ethanol extract is a composition for preventing and treating kidney cancer, characterized in that the drying and concentrated under reduced pressure conditions 45 ℃.
5. The composition for preventing and treating kidney cancer according to any one of claims 1 to 4, wherein the composition comprises a pharmaceutically acceptable carrier or diluent.
6. The composition for preventing and treating kidney cancer according to any one of claims 1 to 4, wherein the kidney cancer is kind cell cell cancer.
7. A cosmetic composition for preventing kidney cancer, comprising a civil extract of cosmetics containing an acceptable cosmetic supplement additive as an active ingredient.

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