WO2012134250A2 - Composition for renal cancer treatment and composition for cosmetic containing nardostachyos rhizoma extract - Google Patents

Composition for renal cancer treatment and composition for cosmetic containing nardostachyos rhizoma extract Download PDF

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WO2012134250A2
WO2012134250A2 PCT/KR2012/002465 KR2012002465W WO2012134250A2 WO 2012134250 A2 WO2012134250 A2 WO 2012134250A2 KR 2012002465 W KR2012002465 W KR 2012002465W WO 2012134250 A2 WO2012134250 A2 WO 2012134250A2
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composition
extract
kidney cancer
present
persimmon
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PCT/KR2012/002465
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French (fr)
Korean (ko)
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WO2012134250A3 (en
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황성연
정경채
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주식회사 한국전통의학연구소
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/84Valerianaceae (Valerian family), e.g. valerian
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones

Definitions

  • the present invention relates to a novel use of persimmon extract. Specifically, the present invention relates to a therapeutic composition and a cosmetic composition containing as an active ingredient extracts persimmon extract showing excellent preventive or therapeutic efficacy against kidney cancer.
  • 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.
  • kidney 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.
  • 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.
  • tumors tend to spread in veins, and metastasis to other organs is likely to occur.
  • VHL von-Hippel-Lindau
  • 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.
  • a sense songhyang (Nardostachytis Rhizoma) is used as the root of a perennial plant of a sense songhyang (nardostachys Batalin chinensis) stomach pain, stomach cramps, thoracoabdominal coupon, neurogenic gastrointestinal disorders, vomiting, headache, or the like, respectively.
  • Nardostachys jatamansi (NJ) has been used extensively as a tonic, irritant and anticonvulsant in several Asian countries, as well as to treat epilepsy, pathological excitement, palpitations and spasms (Bagchi, A., et al. , Planta Med., 57, 9697 (1991)).
  • Perennial perennial persimmon roots include 1-aristol-2-one, nardostachone, 1,8,9,10-tetrad-hydroaristolan-2- components such as one) are contained.
  • the self-acting effect of the sensational sensation relieves pain caused by abdominal pain and chest abdominal pain, and is also applied to neuropathic pain and headache. Wash each affected area by adding water to it. It acts on the central nervous system and has a calming effect, such as heel chogeun ( ⁇ ⁇ ).
  • heart rate control is remarkable, and smooth muscle spasm of the bronchus, small intestine, large intestine, and uterus is released. Minor but also antibacterial.
  • the present inventors completed the present invention by confirming that the extract of the extract of persimmon can effectively kill the kidney cancer cells during the study of herbal medicine for the extract.
  • the present invention provides a composition for the prevention and treatment of kidney cancer comprising an active ingredient extracted from Nardostachyos Rhizoma with an organic solvent.
  • the organic solvent is preferably ethanol.
  • the present invention provides a cosmetic composition for preventing kidney cancer, containing the sensitized ethanol extract containing cosmetically acceptable cosmetic auxiliary additives as an active ingredient.
  • the present invention provides a composition for the prevention and treatment of kidney cancer containing Nardostachyos Rhizoma ethanol extract as an active ingredient.
  • the composition of the present invention contains the extract of Persimmon as an active ingredient, and may further include a pharmaceutically acceptable carrier or diluent.
  • 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%.
  • the kidney cancer is preferably renal cell cancer.
  • the persimmon extract of the present invention is possible at any site of persimmon, but is preferably extracted from the root.
  • 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.
  • ethanol 1 to 5 times, preferably 3 times, 95% ethanol is added to the dried persimmon fragrance or powder, and 10 to 100 hours, preferably 15 at a temperature of 20 to 100 ° C, preferably 40 to 60 ° C.
  • 10 to 100 hours preferably 15 at a temperature of 20 to 100 ° C, preferably 40 to 60 ° C.
  • To 40 hours, more preferably, for 24 hours after extraction can be filtered to prepare an ethanol extract of persimmon flavor.
  • the filtrate obtained by filtering the extract may be concentrated under reduced pressure.
  • 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. .
  • Formulation for pharmacological extracts of the present invention can be administered orally during 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.
  • liquid preparations for oral use include suspensions, solvents, emulsions, and syrups.
  • various excipients may be included, such as wetting sweeteners, fragrances, preservatives and the like.
  • 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, Poly
  • 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.
  • solid preparations include tablets, pills, powders, granules, capsules, and the like.
  • excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included.
  • preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • 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.
  • base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • 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 water for injection 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.
  • 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.
  • the ethanol extract of the fennel flavor kills 96.8% of ACHN kidney cancer cells at 100 ⁇ g / ml and the EC 50 (half maximal effective concentration) is 30.0 ⁇ g / ml.
  • prevention means any action that inhibits or delays the development of kidney cancer by administration of the composition.
  • treatment means any action that improves or advantageously changes the symptoms of kidney cancer by administration of the composition.
  • Persimmon 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.
  • 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.
  • 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.
  • the extract can perform an additional fractionation process.
  • 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.
  • 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.
  • the term "pharmaceutically acceptable salts” means salts derived from pharmacologically or physiologically acceptable inorganic acids, organic acids and bases.
  • suitable acids 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.
  • 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.
  • 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.
  • the composition may include a substance having a therapeutic activity against kidney cancer used alone or used in the past.
  • 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.
  • a composition comprising the extract of Persimmon Fragrance of the present invention, the kidney cancer mentioned above can be effectively prevented and treated.
  • the composition of the present invention can be administered in parallel with existing kidney cancer treatments.
  • 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.
  • the composition may be administered by any device in which the active agent may migrate to the target cell.
  • composition of the present invention is administered in a pharmaceutically effective amount.
  • 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.
  • 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.
  • 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.
  • 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 sex, age, health condition, diet, time of administration, method of administration, drug mixture, the condition of the patient, and the extent of the onset of neurological disease.
  • the present invention provides a cosmetic composition for preventing kidney cancer containing Nardostachyos Rhizoma ethanol extract containing a cosmetically acceptable cosmetic supplement 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.
  • the present invention when the extract of the present invention is added to the ACHN kidney cancer cells, it was confirmed that the ACHN cells are killed (see Fig. 1 ). Thus, it can be seen that the extract of Seng Songhyang has a therapeutic effect on kidney cancer.
  • the present invention was completed by preparing a cosmetic composition for preventing kidney cancer containing the extract of Persimmon as an active ingredient (see Preparation Example 2 ).
  • the 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.
  • ACHN cells which are human kidney cancer cells
  • 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.
  • 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.
  • FBS fetal bovine serum, fetal bovine serum
  • HEPES 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid
  • Example 1 inhibits the growth of ACHN cells which are kidney cancer cells. Specifically, 6.0 ⁇ 10 per well in a 96 well plate 3 The saenghyang incense dissolved in dimethyl sulfoxide (DMSO) after seeding and incubating for 24 hours in ACHN kidney cancer cells.
  • DMSO dimethyl sulfoxide
  • kidney cancer cells were killed at 5.0% at 6.25 ⁇ g / ml, 11.8% at 12.5 ⁇ g / ml, 42.0% at 25 ⁇ g / ml, 72.5% at 50 ⁇ g / ml, and 96.8% at 100 ⁇ g / ml.
  • an EC 50 half maximal effective concentration
  • Table 1 describes the relative cell numbers of kidney cancer cells after 48 hours according to the concentrations of each cheongsam, based on the number of survival rates of the kidney cancer cells of the control group which was not treated with saccharin.
  • the extract of Persimmon Fructus of the present invention has excellent ACHN cell killing activity and further demonstrates that it has therapeutic and prophylactic activity for kidney cancer.
  • the sensational fragrance used in the present invention was widely used as a medicinal herb, so it was determined that there was no problem in stability, but the oral administration and the intraperitoneal administration were performed by conducting toxicological tests.
  • 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 in suspension of the persimmon extract of Example 1 of the present invention in 0.5% methylcellulose solution, respectively. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed, and autopsy was performed to observe abdominal and thoracic organ abnormalities.
  • the present inventors have confirmed that the renal cancer treatment effect of the extract persimmon sensation through the above embodiment was prepared to prepare a renal cancer treatment containing the extract as an active ingredient as follows.
  • the preparation of the following therapeutic agents can be used for the application of not only therapeutic agents but also cosmetic compositions.
  • the present inventors have confirmed that the extract persimmon extract is excellent in the treatment of kidney cancer through the above embodiment to prepare a cosmetic composition containing it as an active ingredient as follows.
  • the nourishing cream was prepared by a conventional method according to the composition described below.
  • Glycerin 4.0%, Polyvinyl Alcohol 15.0%, Hyaluronic Acid Extract 5.0%, Beta Glucan 7.0%, Allantoin 0.1%, Honey Extract 0.5%, Nonyl Phenyl Ether 0.4%, Polysorbate 60 1.2%, Ethanol 6.0%, Purified Water
  • Ointments were prepared in a conventional manner according to the compositions described below.

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Abstract

The present invention relates to a novel use of a nardostachyos rhizoma extract, and more specifically, to a composition for renal cancer treatment and a composition for cosmetics containing, as an active ingredient, a nardostachyos rhizome extract having excellent preventative and treatment effects on renal cancer. The nardostachyos rhizoma extract, according to the present invention, suppresses growth of and induces apoptosis of renal cancer cells, and therefore can be used effectively for renal cancer treatment and prevention.

Description

감송향 추출물을 포함하는 신장암 치료용 조성물 및 화장료 조성물Composition and cosmetic composition for treatment of kidney cancer comprising extract
본 발명은 감송향 추출물의 신규한 용도에 관한 것이다. 구체적으로, 본 발명은 신장암에 대해 탁월한 예방 또는 치료 효능을 나타내는 감송향 추출물을 유효성분으로 함유하는 치료용 조성물 및 화장료 조성물에 관한 것이다.The present invention relates to a novel use of persimmon extract. Specifically, the present invention relates to a therapeutic composition and a cosmetic composition containing as an active ingredient extracts persimmon extract showing excellent preventive or therapeutic efficacy against kidney cancer.
현대인의 주요 질환 중에서, 암의 치료방법과 진단방법에 관한 연구는 발병빈도가 높은 폐암, 간암, 위암 등을 중심으로 진행되고 있다.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.
신장은 혈액을 여과하여 소변을 생성함으로써 생체 내의 노폐물을 체외로 배설하는 역할을 갖는 중요한 비뇨기계 기관이다. 또한, 동시에 혈압을 조절하는 안지오텐신, 적혈구 조혈 인자인 에리스로포이에틴 등의 호르몬을 생산하는 중요한 내분비 기관이기도 하다. 신장에 발생하는 종양에는, 성인에게서 발생하는 신세포암(kindney cell cancer)과 소아에게서 발생하는 윌름스(Wilms) 종양, 및 보기 드문 종양으로서 육종(sarcoma)이 있지만, 이하에서는 가장 발생률이 높은 악성 종양인 신세포암을 신장암이라고 부른다. 신장암의 발생 빈도는 인구 10만명당 2.5명 정도이고, 남녀비는 2 내지 3:1로 남성에게 많은 경향이 있다. 비뇨기과계 악성 종양 중에서는, 전립선암, 방광암에 이어서 많은 종양이다.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.
신장암의 위험 인자로서는 유전학적 요인도 알려져 있지만, 일반적으로는 흡연, 과도한 지방 섭취 등을 들 수 있다. 또한, 장기간 투석을 받고 있는 환자에게서 상기 종양의 발생률이 높은 것으로 알려져 있다. 신장암에서는 종양의 최대 직경이 5 ㎝ 이하인 경우에는 어떠한 자각 증상이 있는 경우가 드물고, 검진시의 CT 스캔 등에 의해 발견되는 경우가 많다. 크기가 큰 종양에서는 혈뇨, 복부 종양, 동통 등이 나타날 수 있다. 또한, 전신적 증상으로서 발열, 체중 감소, 빈혈 등을 초래하는 경우가 있으며, 드물게는 내분비 인자에 의해 적혈구 증다증이나 고혈압, 고칼슘혈증 등이 발생되는 경우가 있다. 한편, 신장암의 하대정맥 내로의 진전에 의해 복부 체표의 정맥의 노장(怒張)이나 정소 정맥류가 발생하는 경우가 있다. 신장암의 약 2할은, 폐나 골 전이로부터 발견된다. 신장암에서는 정맥 중에 종양이 확산되는 경향이 강하고, 다른 장기로의 전이가 발생하기 쉽다.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.
신장암의 검사법으로서는 초음파 검사, CT 검사, 혈관 조영 검사 등이 있지만, 특이적인 생화학 마커는 알려져 있지 않기 때문에 기기를 이용한 검진이 필요로 된다. 또한, 신장암은 병리학적으로 몇 개의 분류가 가능하지만, 그 중 약 90 %를 차지하는 투명 세포형 신장암의 발생 원인은 암 억제 유전자인 VHL(von-Hippel-Lindau) 유전자의 결손인 것으로 알려져 있다. VHL 유전자의 결손은 HIF-α/VHF의 회합 장해를 통해 저산소 유도 유전자군의 전사 활성화를 초래하여, VEGF, TGFβ 등의 유전자의 발현 증강이 발생한다고 알려져 있다.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.
신장암의 주 치료법은 외과 요법이다. 병의 시기에 관계없이, 적출할 수 있는 경우에는 신장의 적출, 또는 신장의 부분적 적출이 가장 일반적이며, 만일 전이가 있다 하더라도 신장의 외과적 적출이 고려되는 경우가 있다. 외과 요법 이외의 방법으로서는 신동맥의 동맥색전술이 있으며, 이 방법은 적출이 불가능한 경우나, 큰 종양을 적출하는 경우에, 수술에 앞서서 시행되는 경우가 있다. 최근 화상 진단 기술의 발전에 의해 신장암은 매우 작은 크기여도 조기에 발견되도록 되어, 초기 암에서의 치료에서는 90% 이상이 치유된다. 그러나, 5 ㎝ 이상의 큰 종양이나 전이가 있는 종양의 치료 성과는 빈약하기 때문에(신장 세포암 전체의 5년 생존율은 약 50 내지 60%), 조기 발견의 중요성이 인식되고 있다.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.
한편, 감송향(Nardostachytis Rhizoma)은 다년생 초본인 감송향(nardostachys chinensis Batalin)의 뿌리로서 위통, 위장경련, 흉복장만, 신경성 위장병, 구토, 두통, 각기 등에 사용된다. 감송향(Nardostachys jatamansi, NJ)은 몇몇 아시아 국가들에서 강장제, 자극제 및 항경련제로 광범위하게 사용되었을 뿐 아니라, 간질, 병적 흥분, 심계 항진 및 경련을 치료하는데도 사용되어 왔다(Bagchi, A., 등, Planta Med., 57, 9697 (1991)). 야타만시산(Jatamansic acid) 및 야타만손 (Jatamansone)과 같은 다양한 세스퀴테르펜들(sesquiterpenes), 리그난, 및 네오리그난이 뿌리에 존재한다(Chatterji, A, 등, National Institute of Science Communication, vol. 5, pp 99-100 (1997); Arora, RB., Indian Council of Medical Research, vol. 51 (1965)).On the other hand, a sense songhyang (Nardostachytis Rhizoma) is used as the root of a perennial plant of a sense songhyang (nardostachys Batalin chinensis) stomach pain, stomach cramps, thoracoabdominal coupon, neurogenic gastrointestinal disorders, vomiting, headache, or the like, respectively. Nardostachys jatamansi (NJ) has been used extensively as a tonic, irritant and anticonvulsant in several Asian countries, as well as to treat epilepsy, pathological excitement, palpitations and spasms (Bagchi, A., et al. , Planta Med., 57, 9697 (1991)). Various sesquiterpenes, lignans, and neolignans such as Jatamansic acid and Jatamansone are present in the roots (Chatterji, A, et al., National Institute of Science Communication, vol. 5, pp 99-100 (1997); Arora, RB., Indian Council of Medical Research, vol. 51 (1965)).
미나리과의 여러해살이풀 감송향 뿌리에는 1-아리스톨렌-2-원(one), 나르도스타촌(nardostachone), 1,8,9,10-테트레이드 하이드로아리스톨란(tetrad-hydroaristolan-2-one) 등의 성분이 함유되어 있다. 감송향의 이기작용은 복통 및 흉복부가 창만하여 일어나는 통증을 완화시키고, 신경성위통과 두통에도 적용된다. 각기(脚氣)에는 물을 넣고 달여서 환부를 세척한다. 중추신경계통에 작용하여 힐초근(尉草根)과 같은 진정작용이 있다. 심장박동이 일정하지 않은 사람에게 심장박동 조절작용이 현저하며, 기관지, 소장, 대장, 자궁 등의 평활근 경련을 풀어준다. 미약하지만 항균작용도 한다. 향성으로 개위성비의 효능이 있기 때문에 위허한으로 인한 완복만통, 식욕부진등의 증을 다스린다. 또한 모든 향료와 합해 향료를 만든다. 감송향의 뿌리를 달인 물은 정신장애, 불면증, 혈액장애 및 순환계 장애에 사용되었으며(Uniyal, MR., 등, J. Res. Indian Med. 4, 83 (1969)), 감송향의 주요한 치료 성분은 향신경 활성 및 중추 신경계(CNS) 효과를 증진시키는 나도시논(nardosinone)이다(Li, p. 등, Journal of Pharmacology Science, 93, 122-125 (2003)). 또한, 감송향의 알콜 추출물은 티오아세트아마이드 유도 간 손상으로부터 쥐를 보호한다(Bagchi, A., 등). 그러나, 아직까지 감송향의 신장암 관련 질환과 관련하여 상기 문헌 어디에도 개시되거나 공개된 바가 없다.Perennial perennial persimmon roots include 1-aristol-2-one, nardostachone, 1,8,9,10-tetrad-hydroaristolan-2- components such as one) are contained. The self-acting effect of the sensational sensation relieves pain caused by abdominal pain and chest abdominal pain, and is also applied to neuropathic pain and headache. Wash each affected area by adding water to it. It acts on the central nervous system and has a calming effect, such as heel chogeun (尉 草根). In people with a constant heart rate, heart rate control is remarkable, and smooth muscle spasm of the bronchus, small intestine, large intestine, and uterus is released. Minor but also antibacterial. Because it is fragrant, it has the effect of false positive effect, so it can alleviate symptoms such as abdominal pain and loss of appetite caused by falsehood. It is also combined with all spices to make spices. The rooted water of the incense incense has been used for mental disorders, insomnia, blood disorders and circulatory disorders (Uniyal, MR., Et al., J. Res. Indian Med. 4, 83 (1969)), the main therapeutic component of the incense incense Is nardosinone that enhances neuronal activity and central nervous system (CNS) effects (Li, p. Et al., Journal of Pharmacology Science, 93, 122-125 (2003)). In addition, the extract of saengsam fragrance protects mice from thioacetamide induced liver damage (Bagchi, A., et al.). However, nothing has been disclosed or disclosed in this document yet in connection with the renal cancer-related diseases of S. aureus.
이에, 본 발명자들은 감송향에 대한 생약 연구를 하던 중, 감송향 추출물이 신장암 세포를 효과적으로 사멸시킬 수 있음을 확인함으로써 본 발명을 완성하였다.Accordingly, the present inventors completed the present invention by confirming that the extract of the extract of persimmon can effectively kill the kidney cancer cells during the study of herbal medicine for the extract.
본 발명의 목적은 감송향 추출물을 유효성분으로 함유하는 신장암 치료용 조성물 및 화장료 조성물을 제공하는 것이다.It is an object of the present invention to provide a composition for treating kidney cancer and a cosmetic composition containing an extract of Persimmon as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 감송향(Nardostachyos Rhizoma)을 유기용매로 추출한 유효성분을 포함하는 신장암 예방 및 치료용 조성물을 제공한다. 상기 유기용매는 에탄올인 것이 바람직하다. In order to achieve the above object, the present invention provides a composition for the prevention and treatment of kidney cancer comprising an active ingredient extracted from Nardostachyos Rhizoma with an organic solvent. The organic solvent is preferably ethanol.
또한, 본 발명은 화장품학적으로 허용 가능한 화장품 보조 첨가제를 포함하는 감송향 에탄올 추출물을 유효성분으로 함유하는 신장암 예방용 화장료 조성물을 제공한다.In another aspect, the present invention provides a cosmetic composition for preventing kidney cancer, containing the sensitized ethanol extract containing cosmetically acceptable cosmetic auxiliary additives as an active ingredient.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 감송향(Nardostachyos Rhizoma) 에탄올 추출물을 유효성분으로 함유하는 신장암 예방 및 치료용 조성물을 제공한다. 본 발명의 조성물은 활성성분으로서 감송향 추출물을 포함하며, 추가적으로 약제학적으로 허용가능한 담체 또는 희석제를 포함할 수 있다.The present invention provides a composition for the prevention and treatment of kidney cancer containing Nardostachyos Rhizoma ethanol extract as an active ingredient. The composition of the present invention contains the extract of Persimmon as an active ingredient, and may further include a pharmaceutically acceptable carrier or diluent.
본 발명의 신장암 예방 및 치료용 조성물에 있어서, 상기 에탄올 추출물은 50℃에서 24시간 동안 추출되는 것이 바람직하고, 이때 상기 에탄올 추출물은 45℃ 감압 조건에서 건조 및 농축되는 것이 보다 바람직하며, 상기 에탄올은 95%인 것이 가장 바람직하다.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%.
또한, 본 발명의 신장암 예방 및 치료용 조성물에 있어서, 상기 신장암은 신세포암(kindney cell cancer)인 것이 바람직하다.In addition, in the composition for preventing and treating kidney cancer of the present invention, the kidney cancer is preferably renal cell cancer.
본 발명의 감송향 추출물은 감송향의 어느 부위에서도 가능하지만, 바람직하게는 뿌리로부터 추출되는 것이 바람직하다. 그리고 추출용액은 물 또는 유기용매로 추출하여 얻을 수 있는데, 유기용매로는 저급 알콜, 아세톤, 클로로포름, 메틸렌클로라이드, 에테르, 에틸아세테이트, 헥산 등을 예시할 수 있다. 저급알콜로는 메탄올, 에탄올, 프로판올 및 부탄올을 예시할 수 있으며, 에탄올이 가장 바람직하다.The persimmon extract of the present invention is possible at any site of persimmon, but is preferably extracted from the root. 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.
구체적으로, 감송향 건조물 또는 분말에 1 내지 5 배, 바람직하게는 3배의 95% 에탄올을 첨가하고 20 내지 100℃, 바람직하게는 40 내지 60℃의 온도에서 10 내지 100시간, 바람직하게는 15 내지 40시간, 더욱 바람직하게는 24시간 동안 추출한 후 여과하여 감송향의 에탄올 추출물을 제조할 수 있다. 바람직하게는 상기 추출액을 여과하여 얻어진 여액을 감압농축하여 제조할 수 있다. 상기 추출방법들에서 추출 공정은 필요에 따라 2회 이상 반복하여 실시할 수 있으며, 여과 후 얻어진 추출물을 동결 건조 또는 감압건조시켜 분말 형태로 만들 수도 있다.Specifically, 1 to 5 times, preferably 3 times, 95% ethanol is added to the dried persimmon fragrance 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, for 24 hours after extraction can be filtered to prepare an ethanol extract of persimmon 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.
상기 "약제학적으로 허용가능한 담체"는 신체의 한 기관 또는 부분으로부터 신체의 다른 기관 또는 부분으로 활성 성분을 수송하는 역할을 하는 액체 또는 고체 충진제, 희석제, 부형제 또는 용매와 같은 약제학적으로 허용되는 물질, 조성물 또는 운반체(vehicle)를 의미한다.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.
본 발명의 신장암 치료용 조성물은 유효성분과 함께 추가로 약제학적으로 허용되는 1종 이상의 담체를 첨가하여 약제로 제조할 수 있다. 상기 담체로는 식염수, 완충 식염수, 물, 글리세롤 및 에탄올 등이 있으나 이에 한정되지 않으며, 당해 기술 분야에 알려진 적합한 제제(Remingtons's Pharmaceutical Science(최근판), Mack Publishing Company, Easton PA)를 모두 사용 가능하다.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. .
본 발명의 감송향 추출물을 약제화하기 위한 제제는 임상 투여시에 경구로 투여가 가능하며 일반적인 의약품 제제의 형태로 사용될 수 있으며, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 경구를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드, 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제 감미제, 방향제, 보존제 등이 포함될 수 있다.Formulation for pharmacological extracts of the present invention can be administered orally during 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.
또한, 본 발명의 조성물에 추가될 수 있는 생약재는 약학적으로 허용되는 임의의 생약재일 수 있으며, 예를 들면, 고본(Angelicae tenuissimae Radix), 천마(Gastrodiae Rhizoma), 시호(Bapleuri Radix), 당귀(Angelicae gigantis Radix), 도인(Persicae Semen), 계지(Cinnamomi Ramulus), 대황(Rhei Rhizoma), 감초(Glycyrrhizae Radix), 천궁(Cnidii Rhizoma), 진피(Aurantii nobilis Pericarpium), 택사(Alismatis Rhizoma), 황련(Coptidis Rhizoma), 황금(Scutellariae Radix), 복령(Hoelen), 작약(Paeoniae Radix), 백출(Atractylodis Rhizoma alba), 황백(Phellodendri Cortex), 치자(Gardeniae Fructus), 반하(Pinelliae Tuber), 조구등(Uncaria Ramuluset Uncus), 지실(Ponciri Fructus), 인삼(Gingseng), 맥문동(Liriopis Tuber), 원지(Polygalae Radix), 석창포(Acori graminei Rhizoma), 창출(Atractylodis Rhizoma alba), 감국(Chrysanthemi Flos), 방풍(Ledebouriellae Radix), 생강(Zingiberis Rhizoma crudus), 망초(Natrii sulfas), 대조(Zizyphi Fructus), 단삼(Salviae Radix), 목단피(Mautan Radicis Cortex), 지황(Rehmanniae Radix), 박하(Menthae Herba), 산약(Dioscoreae Rhizoma), 저령(Polyporus), 하수오(Polygonimultiflori Radix), 구자(Allii tuberosi Semen), 결명자(Cassiae Semen), 구기자(Lycii Fructus), 독활(Araliae cordatae Radix), 두충(Eucommiae Cortex), 백화사설초(Hedyotis Herba), 삼백초(Saururus Herba), 인진(Artemisiaecapillaris Herba), 지모(Anemarrhenae Rhizoma), 홍화(Carthami Flos), 황기(Astragali Radix), 석송자(Lycopodium), 은행잎(Ginkgonis Folium), 황정(Polygonati Rhizoma), 연자육(Nelumbinis Semen), 용골(Fossilia ossis Mastodi), 지골피(Lycii radicis Cortex), 우슬(Achyranthis Radix), 숙지황(Rehmanniae Radix preparata), 흑임자(Perillae Semen), 백자인(Thujae Semen), 맥아(Hordei Fructus germinatus), 토사자(Cuscutae Semen), 파극천(Morindae Radix), 해송(Pini koraiensis Radix) 등을 단독으로 또는 배합하여 사용할 수 있다.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, Minthae 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.
본 발명의 조성물은 실제 임상 투여시에 비경구의 여러 가지 제형으로 투여될 수 있는데, 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드, 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제 감미제, 방향제, 보존제 등이 포함될 수 있다. 구체적으로, 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. 또한 치료제의 효능 증진을 위해 칼슘이나 비타민 D3를 첨가할 수 있다. 이러한 조성물은 단위-용량(1회분) 또는 다중-용량(수 회분) 용기, 예를 들면, 밀봉된 앰풀 및 바이알에 제시될 수 있고, 사용 직전에 멸균성 액상 담체, 예를 들면, 주사용 수의 부가 만을 요구하는 동결-건조 조건 하에 저장할 수 있다. 즉석의 주사 용제 및 현탁제는 멸균성 산제, 과립제 및 정제로부터 제조할 수 있다.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 water for injection 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.
본 발명의 제제는 대상의 연령, 성별, 상태, 체내에서 활성 성분의 흡수도, 불활성율 및 배설속도, 병용되는 약물에 따라 달리 적용될 수 있다. 본 발명은 또한 투약 단위의 제형들을 포함한다. 제형은 개별 투약 형태, 예를 들면 정제, 피복 정제, 캡슐제, 환제, 좌약 및 앰플제로 존재하고, 약제 중 유효 화합물의 함량은 개별 투약량의 분율 또는 배수에 해당한다. 투약 단위는, 예를 들면 개별 투여량의 1, 2, 3 또는 4배로, 또는 1/2, 1/3 또는 1/4배를 함유할 수 있다. 개별 투여량은 바람직하기로는 유효 화합물이 1회에 투여되는 양을 함유하며, 이는 통상 1일 투여량의 전부, 1/2, 1/3 또는 1/4배에 해당한다.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.
본 발명에서 용어, "추출물(extract)"은 천연물로부터 분리된 활성성분을 의미한다. 추출물은 물, 유기용매, 또는 이의 혼합용매를 이용하는 추출과정으로 획득할 수 있으며, 추출액, 이의 건조 분말 또는 이를 이용하여 제형화된 모든 형태를 포함한다.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.
본 발명의 구체적인 실시에서, 감송향의 에탄올 추출물은 ACHN 신장암 세포를 100 ㎍/㎖에서 96.8% 사멸시키고, EC50(half maximal effective concentration)은 30.0 ㎍/㎖이었다. 상기 결과는 본 발명의 감송향 추출물이 우수한 ACHN 신장암 세포의 사멸 활성을 가지며, 나아가 신장암 치료 및 예방 활성을 가진다는 것을 입증하는 것이다.In a specific embodiment of the present invention, the ethanol extract of the fennel flavor kills 96.8% of ACHN kidney cancer cells at 100 μg / ml and the EC 50 (half maximal effective concentration) is 30.0 μg / ml. The above results demonstrate that the extract of Persimmon Fructus of the present invention has excellent killing activity of ACHN kidney cancer cells, and further has renal cancer treatment and prophylactic activity.
본 발명에서 용어, "예방"이란 조성물의 투여로 신장암 발병을 억제 또는 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" means any action that inhibits or delays the development of kidney cancer by administration of the composition.
본 발명에서 용어, "치료"란 조성물의 투여로 신장암의 증세가 호전되거나 이롭게 변경하는 모든 행위를 의미한다.As used herein, the term "treatment" means any action that improves or advantageously changes the symptoms of kidney cancer by administration of the composition.
본 발명에서 감송향 추출물은 물, 유기 용매, 또는 이의 혼합 용매를 사용하여 추출하여 사용할 수 있다. 바람직하게는 유기 용매, 특히 에탄올을 사용하여 추출한다. 추출한 액은 바로 사용하거나 또는 농축 및/또는 건조하여 사용할 수 있다. 유기용매를 사용하여 추출하는 경우, 메탄올, 에탄올, 이소프로판올, 부탄올, 에틸렌, 아세톤, 헥산, 에테르, 클로로포름, 에틸아세테이트, 부틸아세테이트, 디클로로메탄, N, N-디메틸포름아미드(DMF), 디메틸설폭사이드(DMSO), 1,3-부틸렌글리콜, 프로필렌글리콜 또는 이들의 혼합용매인 유기용매를 사용하며 생약의 유효 성분이 파괴되지 않거나 최소화된 조건에서 실온 또는 가온하여 추출할 수 있다. 추출하는 유기용매에 따라 약제의 유효성분의 추출정도와 손실정도가 차이가 날 수 있으므로, 알맞은 유기용매를 선택하여 사용하도록 한다. 추출 방법은 특별히 제한되지 않고, 예를 들어 냉침 추출, 초음파 추출, 환류 냉각 추출 등이 있다. 여과는 추출액으로부터 부유하는 고체 입자를 제거하는 과정으로, 면, 나일론 등을 이용하여 입자를 걸러내거나 한외여과, 냉동여과법, 원심분리법 등을 사용할 수 있으나 이에 제한되지 않는다.Persimmon 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.
추출액의 농축에는 감압농축, 역삼투압 농축 등의 방법이 사용될 수 있다. 농축 후 건조 단계는 동결건조, 진공건조, 열풍건조, 분무건조, 감압건조, 포말건조, 고주파건조, 적외선건조 등을 포함하나 이에 제한되지 않는다. 경우에 따라, 최종 건조된 추출물을 분쇄하는 공정을 추가할 수 있다.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.
또한, 상기 추출물은 추가의 분획 공정을 수행할 수 있다. 바람직하게는 상기 추출물을 증류수에 현탁시켜 비극성 유기 용매, 예를 들어, 헥산, 에테로, 디클로로메탄, 클로로포름, 에틸아세테이드 또는 이들의 혼합 용매로 비극성용매 가용층을 추출, 분리하여 수득하도록 하고, 이를 농축 및/또는 건조하여 사용할 수 있다.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.
본 발명에서, 용어 "약제학적으로 허용가능한 염"이란 약리학적 또는 생리학적으로 허용되는 무기산, 유기산 및 염기로부터 유도된 염을 의미한다. 적합한 산의 예로는 염산, 브롬산, 황산, 질산, 과염소산, 푸마르산, 말레산, 인산, 글리콜산, 락트산, 살리실산, 숙신산, 톨루엔-p-설폰산, 타르타르산, 아세트산, 시트르산, 메탄설폰산, 포름산, 벤조산, 말론산, 나프탈렌-2-설폰산, 벤젠설폰산 등을 포함할 수 있다. 적합한 염기로부터 유도된 염은 알칼리 금속, 예들 들어, 나트륨, 알칼리토금속, 예들 들어, 마그네슘, 암모늄 등을 포함할 수 있다.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 acids 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.
본 발명의 신장암 질환 예방 및 치료용 약학조성물은 조성물 총 중량에 대하여 상기 추출물 또는 화합물을 0.1 내지 50 중량%로 포함한다. 또한, 상기 조성물은 약효를 증가시키지는 않으나 약재 조성물에 통상 사용되어 냄새, 맛, 시각 등을 향상시킬 수 있는 추가성분을 포함할 수 있다. 또한, 상기 조성물은 비타민 B1, B2, B6, C, E, 니아신, 카르니친, 베타인, 엽산 판토텐산, 비오틴, 아연, 철, 칼슘, 크롬, 마그네슘, 이들의 혼합물 등의 무기, 유기 첨가물들을 추가로 포함할 수 있다. 또한, 상기 조성물은 단독 사용하거나 기존 사용되어진 신장암에 대한 치료 활성을 가지는 물질을 포함할 수 있다.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.
본 발명에서 용어, "환자"는 신장암 및 이의 직, 간접적 원인에 의해 유발된 질환을 가지고, 본 발명의 조성물을 투여에 의하여 증상이 호전될 수 있는 인간과 말, 양, 돼지, 염소, 낙타, 영양, 개 등의 동물을 의미한다. 본 발명의 감송향 추출물을 포함하는 조성물을 환자에게 투여함으로써, 상기에서 언급한 신장암을 효과적으로 예방 및 치료할 수 있다. 본 발명의 조성물을 기존의 신장암 치료제와 병행하여 투여할 수 있다.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 the extract of Persimmon Fragrance of the present invention, the kidney cancer mentioned above can be effectively prevented and treated. The composition of the present invention can be administered in parallel with existing kidney cancer treatments.
본 발명에서 용어, "투여"는 어떠한 적절한 방법으로 환자에게 소정의 물질을 도입하는 것을 의미하며, 본 발명의 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 경구 또는 비경구 투여될 수 있다. 또한, 조성물은 활성 물질이 표적 세포로 이동할 수 있는 임의의 장치에 의해 투여될 수 있다.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.
본 발명의 조성물은 약제학적으로 유효한 양으로 투여한다. 본 발명에서 용어, "약제학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 환자의 성병, 연령, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있다. 단일 또는 다중 투여될 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 당업자에 의해 용이하게 결정될 수 있다. 본 발명의 제조 방법에 따라 제조된 추출물 또는 화합물을 포함하는 조성물의 투여방법은 경구투여 또는 정맥투여가 바람직하고, 일반적으로 그 유효 용량은 경구투여인 경우에는 보통 성인을 기준으로 1회에 1 내지 500 ㎎/㎏이 바람직하며, 정맥투여인 경우에는 1 내지 100 ㎎/㎏이 바람직하며, 하루 2-3 회 투여될 수 있다. 특정 환자에 대한 투여용량 수준은 성별, 연령, 건강상태, 식이, 투여시간, 투여방법, 약제혼합, 환자의 상태 및 신경 질환의 발병 정도에 따라 변화될 수 있다. 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 sex, age, health condition, diet, time of administration, method of administration, drug mixture, the condition of the patient, and the extent of the onset of neurological disease.
또한, 본 발명은 화장품학적으로 허용 가능한 화장품 보조 첨가제를 포함하는 감송향(Nardostachyos Rhizoma) 에탄올 추출물을 유효성분으로 함유하는 신장암 예방용 화장료 조성물을 제공한다.In another aspect, the present invention provides a cosmetic composition for preventing kidney cancer containing Nardostachyos Rhizoma ethanol extract containing a cosmetically acceptable cosmetic supplement additive as an active ingredient.
본 발명의 화장료 조성물은 에탄올 추출물을 총중량에 대해 0.01 내지 10중량% 포함하는 것이 바람직하며, 0.1~1중량%로 포함하는 것이 더욱 바람직하다. 본 발명의 화장료 조성물은 그 제형에 있어서 특별히 한정되는 바는 없으며, 예를 들어 유연화장수, 영양화장수, 마사지크림, 영양크림, 팩, 젤 또는 피부 점착타입의 화장료 제형을 가질 수 있으며, 또한 로션, 연고, 젤, 크림, 패취 또는 분무제와 같은 경피투여형의 제형일 수 있다.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.
본 발명에서는 ACHN 신장암 세포에 본 발명의 감송향 추출물을 투입한 결과, ACHN 세포가 사멸하는 것을 확인하였다(도 1 참조). 따라서, 감송향 추출물이 신장암 치료 효과가 있다는 것을 새롭게 알 수 있다. 이에, 본 발명에서는 감송향 추출물을 유효성분으로 함유하는 신장암 예방용 화장료 조성물을 제조함으로써 본 발명을 완성하였다(제조예 2 참조).In the present invention, when the extract of the present invention is added to the ACHN kidney cancer cells, it was confirmed that the ACHN cells are killed (see Fig. 1 ). Thus, it can be seen that the extract of Seng Songhyang has a therapeutic effect on kidney cancer. Thus, in the present invention, the present invention was completed by preparing a cosmetic composition for preventing kidney cancer containing the extract of Persimmon as an active ingredient (see Preparation Example 2 ).
상기에서 살펴본 바와 같이, 본 발명의 감송향 추출물은 신장암 세포의 성장을 억제하고 세포사멸을 유도한다. 따라서 본 발명에 따른 신장암 치료용 조성물은 신장암 환자의 치료에 매우 효과적일 것이다.As described above, the extract of the present invention, the 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.
도 1은 인간 신장암 세포인 ACHN 세포에서 감송향의 도입이 신장암 세포의 성장에 미치는 영향을 알아보기 위한 Alamar Blue 분석 결과이고, 이때 X축은 감송향 추출물의 농도이고, Y축은 생존한 인간 신장암 세포의 생존율을 나타낸다. 1 is a result of the Alamar Blue analysis to determine the effect of the introduction of the incidence in the renal cancer cells of ACHN cells, human kidney cancer cells, wherein the X-axis is the concentration of the extract of the persimmon, Y-axis is the surviving human kidney The survival rate of cancer cells is shown.
이하, 본 발명을 하기 실시예에 의거하여 보다 상세하게 설명하고자 한다. 단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐, 본 발명은 하기 실시예에 의해 한정되는 것이 아니고, 본 발명의 기술적 사상을 벗어나지 않는 범위 내에서 치환 및 균등한 타 실시예로 변경할 수 있음은 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 있어서 명백할 것이다.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.
<실시예 1> 감송향 추출물의 제조Example 1 Preparation of Persimmon Extract
서울 약재상에서 구입한 감송향(중국산) 3 ㎏을 음지 및 실온에서 5일간 건조하고 분쇄하였다. 상기 분쇄된 감송향을 95% 에탄올(ethanol) 30 ℓ에 침지시키고 50℃에서 24시간 동안 추출하였다. 이것을 여과지를 통하여 여과한 후 45℃ 감압 조건에서 건조 및 농축하여 총 추출물 511 g을 수득하고, -20℃에서 보관하였다.3 kg of Gamsong (Chinese) purchased from Seoul medicinal herb was dried and ground for 5 days at the shade and room temperature. The ground persimmon 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 511 g of the total extract, which was stored at −20 ° C.
<실시예 2> 감송향 추출물이 신장암 세포의 성장에 미치는 영향Example 2 Effect of Persimmon Extract on Growth of Kidney Cancer Cells
상기 실시예 1에서 추출한 감송향 추출물이 신장암 세포의 성장에 미치는 영향을 알아보기 위하여, 인간 신장암 세포인 ACHN 세포에 감송향의 에탄올 추출물을 48시간 처리하고 Alamar Blue 분석을 시행하였다. Alamar Blue 분석은 MTT 분석의 변형된 형태인데, 특정 효소에 의해서 분해되는 화합물을 살아있는 세포에 처리한 후 화합물이 분해되면서 나오는 생성물의 형광 세기를 측정함으로써 약물을 처리한 후 살아있는 세포의 상대적인 숫자를 알아내는 실험방법이다. 하기에서 보다 상세히 설명한다.In order to determine the effect of the extract from Example 1 on the growth of kidney cancer cells, ACHN cells, which are human kidney cancer cells, were treated for 48 hours with ethanol extracts of the extracts of the kidney 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> 인간 신장암 세포주의 준비 및 처리<2-1> Preparation and Treatment of Human Kidney Cancer Cell Line
본 발명에 사용된 신장암 세포주인 ACHN 세포는 한국세포주은행(Korean Cell Line Bank, KCLB)으로부터 분양받아 실험에 이용하였다. 구체적으로, ACHN 신장암 세포를 10% FBS(fetal bovine serum, 소태아혈청)(Welgene)와 25 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)를 포함하는 MEM(Minimum essential medium) 배지에서 계대배양하였다.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> ACHN 신장암 세포의 세포 성장 억제 측정<2-2> Cell growth inhibition measurement of ACHN kidney cancer cells
상기 실시예 1에서 추출한 감송향 추출물이 신장암 세포인 ACHN 세포에 대하여 성장을 억제시키는 효과를 확인하였다. 구체적으로, 96 웰 플레이트에 각 웰 당 6.0ㅧ 103 개의 ACHN 신장암 세포를 주입(seeding)하고 24시간 동안 배양한 후, DMSO(Dimethyl sulfoxide)에 녹인 상기 감송향 에탄올 추출물을 각각 0 내지 100 ㎍/㎖ 농도(구체적으로, 각각 0, 3.125, 6.25, 12.5, 25, 50 및 100 ㎍/㎖ 농도)로 48시간 동안 처리하였을 때, 세포 성장을 저해하는 정도를 확인하였다(표 1). 각 농도의 추출물을 처리한 후, 96-웰 플레이트에서 각 웰에 채워진 0.2 ㎖의 세포 배양액에 20 ㎕의 Alamar Blue 시약을 첨가한 후 플레이트를 인큐베이터에서 2시간 동안 배양하였다. 각 웰의 세포를 고르게 반응시키기 위하여 플레이트를 천천히 흔들고, 544 ㎚의 파장에서 조사광을 조사하면서 590 ㎚에서 형광의 세기를 형광광도계(Fluorescence Microplate Reader; Molecular Devices Corp.)로 측정하였고, 신장암 세포의 생존율을 도 1에 나타내었다.It was confirmed that the extract of the extract of Example 1 in Example 1 inhibits the growth of ACHN cells which are kidney cancer cells. Specifically, 6.0 × 10 per well in a 96 well plate3 The saenghyang incense dissolved in dimethyl sulfoxide (DMSO) after seeding and incubating for 24 hours in ACHN kidney cancer cells When the ethanol extract was treated at concentrations of 0 to 100 μg / ml (specifically, concentrations of 0, 3.125, 6.25, 12.5, 25, 50 and 100 μg / ml, respectively) for 48 hours, the degree of inhibition of cell growth was confirmed. (Table 1). After treatment of 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.
표 1
감송향 농도 신장암 세포 생존율 (평균) 표준편차
0 ㎍/㎖ 1.000 0.000
3.125 ㎍/㎖ 1.033 0.017
6.25 ㎍/㎖ 0.950 0.020
12.5 ㎍/㎖ 0.882 0.005
25 ㎍/㎖ 0.580 0.037
50 ㎍/㎖ 0.275 0.023
100 ㎍/㎖ 0.032 0.003
Table 1
Sensation concentration Kidney Cancer Cell Survival (Average) Standard Deviation
0 μg / ml 1.000 0.000
3.125 μg / ml 1.033 0.017
6.25 μg / ml 0.950 0.020
12.5 μg / ml 0.882 0.005
25 μg / ml 0.580 0.037
50 μg / ml 0.275 0.023
100 μg / ml 0.032 0.003
그 결과, 표 1 및 도 1에서 나타난 바와 같이, 감송향의 처리 농도가 높을수록 신장암 세포의 성장이 감소하였으며, 이로부터 감송향이 신장암 치료 효과를 가짐을 알 수 있었다. 즉, 6.25 ㎍/㎖에서 5.0%, 12.5 ㎍/㎖에서 11.8%, 25 ㎍/㎖에서 42.0%, 50 ㎍/㎖에서 72.5%, 100 ㎍/㎖에서 96.8%로 신장암 세포를 사멸시켰다. 아울러, EC50(half maximal effective concentration)은 30.0 ㎍/㎖로 측정되었다. 상기 표 1에서 감송향을 처리하지 않은 대조군의 신장암 세포의 생존율 수를 1을 기준으로 하여 각각의 감송향 처리 농도에 따른 48시간 후의 신장암 세포의 상대적 세포수를 기재하였다. 이와 같이, 본 발명의 감송향 추출물은 우수한 ACHN 세포 사멸 활성을 가지며, 나아가 신장암 치료 및 예방 활성을 가진다는 것을 입증한다.As a result, as shown in Table 1 and FIG. 1, the growth of kidney cancer cells was decreased as the concentration of the sweet persimmon was higher, and it was found that the sweet persimmon had the effect of treating the renal cancer. That is, kidney cancer cells were killed at 5.0% at 6.25 µg / ml, 11.8% at 12.5 µg / ml, 42.0% at 25 µg / ml, 72.5% at 50 µg / ml, and 96.8% at 100 µg / ml. In addition, an EC 50 (half maximal effective concentration) was measured at 30.0 μg / ml. Table 1 describes the relative cell numbers of kidney cancer cells after 48 hours according to the concentrations of each cheongsam, based on the number of survival rates of the kidney cancer cells of the control group which was not treated with saccharin. As such, the extract of Persimmon Fructus of the present invention has excellent ACHN cell killing activity and further demonstrates that it has therapeutic and prophylactic activity for kidney cancer.
<실시예 3> 감송향 추출물에 의한 급성독성 시험Example 3 Acute Toxicity Test by Persimmon Extract
본 발명에 이용된 감송향은 널리 약재로 이용되고 있어서 안정성에 문제가 없을 것으로 판단하였으나, 경구 투여시 및 복강내 투여시의 독성 실험을 수행하여 이를 확인하고자 하였다.The sensational fragrance used in the present invention was widely used as a medicinal herb, so it was determined that there was no problem in stability, but the oral administration and the intraperitoneal administration were performed by conducting toxicological tests.
6주령의 특정병원부재(SPF) SD계 랫트를 사용하여 급성독성실험을 실시하였다. 군당 2 마리씩의 동물에 본 발명의 실시예 1의 감송향 추출물을 각각 0.5% 메틸셀룰로즈 용액에 현탁하여 5 g/㎏의 용량으로 단회 경구투여하였다. 시험물질 투여후 동물의 폐사여부, 임상증상, 체중변화를 관찰하고 혈액학적 검사와 혈액생화학적검사를 실시하였으며, 부검하여 육안으로 복강장기와 흉강장기의 이상여부를 관찰하였다.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 in suspension of the persimmon extract of Example 1 of the present invention in 0.5% methylcellulose solution, respectively. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed, and autopsy was performed to observe abdominal and thoracic organ abnormalities.
시험결과, 시험물질을 투여한 모든 동물에서 특기할 만한 임상증상이나 폐사된 동물은 없었으며, 체중변화, 혈액검사, 혈액생화학 검사, 부검소견 등에서도 독성변화는 관찰되지 않았다. 이상의 결과 감송향 추출물은 모두 랫트에서 5 g/㎏까지 독성변화를 나타내지 않으며 경구 투여 최소치사량 (LD50)은 5 g/㎏이상인 안전한 물질로 판단되었다.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 biochemical test, autopsy findings, etc. As a result, all of the extracts of Persimmon extract did not show toxicity change up to 5 g / kg in rats, and the minimum lethal dose (LD 50 ) was determined to be a safe substance of 5 g / kg or more.
<제조예 1> 감송향 추출물을 유효성분으로 함유하는 신장암 치료제의 제조<Preparation Example 1> Preparation of a renal cancer therapeutic agent containing the extract of Persimmon
본 발명자들은 상기 실시예를 통해 감송향 추출물의 신장암 치료 효능이 뛰어남을 확인하여 상기 추출물을 유효성분으로 함유하는 신장암 치료제를 하기와 같이 제조하였다. 또한, 하기 치료제의 제조예는 치료제 뿐만 아니라 화장료 조성물의 제조에도 응용하여 사용될 수 있다.The present inventors have confirmed that the renal cancer treatment effect of the extract persimmon sensation through the above embodiment was prepared to prepare a renal cancer treatment 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> 감송향 추출물을 함유하는 연질캅셀(soft gelatin capsules)(중량%)<1-1> Soft gelatin capsules (wt%) containing extract of Persimmon
감송향 추출물 20%, 비타민 C 4.5%, 비타민 D3 0.001%, 황산망간0.1%, 밀납10%, 팜유25%, 홍화씨유30.399%Honey 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> 감송향 추출물을 함유하는 정맥주사용 제제의 제조(중량%) <1-2> Preparation of Intravenous Formulation Containing Persimmon Fragrance Extract (wt%)
감송향 추출물0.2%, 만니톨0.3%, 생리식염수9.5%Persimmon extract 0.2%, mannitol 0.3%, physiological saline 9.5%
<1-3> 감송향 추출물을 함유하는 정제(tablet)(중량%)<1-3> Tablet (Weight%) Containing Persimmon Extract
감송향 추출물 35%, 비타민 C 10%, 비타민 D3 0.001%, 황산망간 0.1%, 결정셀룰로오즈 25.0%, 유당 17.999%, 스테아린산마그네슘 2%Honey extract 35%, vitamin C 10%, vitamin D 3 0.001%, manganese sulfate 0.1%, crystalline cellulose 25.0%, lactose 17.999%, magnesium stearate 2%
<제조예 2> 감송향 추출물을 유효성분으로 함유하는 화장료 조성물의 제조Preparation Example 2 Preparation of Cosmetic Composition Containing Persimmon Scent Extract as an Active Ingredient
본 발명자들은 상기 실시예를 통해 감송향 추출물이 신장암 치료 활성이 뛰어남을 확인하여 이를 유효성분으로 함유하는 화장료 조성물을 하기와 같이 제조하였다.The present inventors have confirmed that the extract persimmon extract is excellent in the treatment of kidney cancer through the above embodiment to prepare a cosmetic composition containing it as an active ingredient as follows.
<2-1> 감송향 추출물을 함유하는 영양화장수(중량%)<2-1> Nutritious Cosmetic Water Containing Persimmon Extract (wt%)
하기 기재된 조성에 따라 통상적인 방법으로 영양화장수를 제조하였다.Nutrients were prepared in a conventional manner according to the composition described below.
글리세린 8.0 %, 부틸렌글리콜 4.0 %, 히알루론산 추출물 5.0 %, 베타글루칸 7.0 %, 카보머 0.1 %, 감송향 추출물 0.05 %, 카프필릭/카프릭 트리글리세라이드 8.0 %, 스쿠알란 5.0 %, 세테아릴 글루코사이드 1.5 %, 소르비탄 스테아레이트 0.4 %, 세테아릴 알콜 1.0 %, 트리에탄올 아민 0.1 %, 정제수 잔량Glycerin 8.0%, Butylene Glycol 4.0%, Hyaluronic Acid Extract 5.0%, Betaglucan 7.0%, Carbomer 0.1%, Persimmon Extract 0.05%, Capric / 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> 감송향 추출물을 함유하는 영양크림(중량%)<2-2> Nutritional Cream Containing Persimmon Extract (wt%)
하기 기재된 조성에 따라 통상적인 방법으로 영양크림을 제조하였다.The nourishing cream was prepared by a conventional method according to the composition described below.
글리세린 3.0 %, 부틸렌글리콜 3.0 %, 유동파라핀 7.0 %, 베타글루칸 7.0 %, 카보머 0.1 %, 감송향 추출물 3.0 %, 카프필릭/카프릭 트리글리세라이드 3.0 %, 스쿠알란 5.0 %, 세테아릴 글루코사이드 1.5 %, 소르비탄 스테아레이트 0.4 %, 폴리솔베이트 60 1.2 %, 트리에탄올 아민 0.1 %, 정제수 잔량Glycerin 3.0%, Butylene Glycol 3.0%, Liquid Paraffin 7.0%, Beta Glucan 7.0%, Carbomer 0.1%, Persimmon Extract 3.0%, Capricic / 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> 감송향 추출물을 함유하는 마시지 크림(중량%)<2-3> Massage cream containing extract of sweet persimmon (wt%)
하기 기재된 조성에 따라 통상적인 방법으로 마사지 크림을 제조하였다.Massage creams were prepared in a conventional manner according to the composition described below.
글리세린 8.0 %, 부틸렌글리콜 4.0 %, 유동파라핀 45.0 %, 베타글루칸 7.0 %, 카보머 0.1 %, 감송향 추출물 1.0 %, 카프필릭/카프릭 트리글리세라이드 3.0 %, 밀납 4.0 %, 세테아릴 글루코사이드 1.5 %, 세스퀴 올레인산 소르비탄 0.9 %, 바세린 3.0 %, 파라핀 1.5 %, 정제수 잔량Glycerin 8.0%, Butylene Glycol 4.0%, Liquid Paraffin 45.0%, Beta Glucan 7.0%, Carbomer 0.1%, Persimmon 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> 감송향 추출물을 함유하는 팩(중량%)<2-4> Pack containing the sweet persimmon extract (wt%)
하기 기재된 조성에 따라 통상적인 방법으로 팩을 제조하였다.Packs were prepared by conventional methods according to the compositions described below.
글리세린 4.0 %, 폴리비닐알콜 15.0 %, 히알루론산 추출물 5.0 %, 베타글루칸 7.0 %, 알란토인 0.1 %, 감송향 추출물 0.5 %, 노닐 페닐에테르 0.4 %, 폴리솔베이트 60 1.2 %, 에탄올 6.0 %, 정제수 잔량Glycerin 4.0%, Polyvinyl Alcohol 15.0%, Hyaluronic Acid Extract 5.0%, Beta Glucan 7.0%, Allantoin 0.1%, Honey Extract 0.5%, Nonyl Phenyl Ether 0.4%, Polysorbate 60 1.2%, Ethanol 6.0%, Purified Water
<2-5> 감송향 추출물을 함유하는 피부외용제 연고(중량%)<2-5> Ophthalmic skin ointment containing weight extract
하기 기재된 조성에 따라 통상적인 방법으로 연고를 제조하였다.Ointments were prepared in a conventional manner according to the compositions described below.
글리세린 8.0 %, 부틸렌글리콜 4.0 %, 유동파라핀 15.0 %, 베타글루칸 7.0 %, 카보머 0.1 %, 감송향 추출물 3.0 %, 스쿠알란 1.0 %, 세테아릴 글루코사이드 1.5 %, 소르비탄 스테아레이트 0.4 %, 세테아릴 알콜 1.0 %, 밀납 4.0 %, 정제수 잔량Glycerin 8.0%, Butylene Glycol 4.0%, Liquid Paraffin 15.0%, Beta Glucan 7.0%, Carbomer 0.1%, Persimmon 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
한편, 본 발명의 구체적 범위는 상기 기술한 실시예 보다는 특허청구범위에 의하여 한정지어지며, 특허청구 범위의 의미와 범위 및 그 등가적 개념으로 도출되는 모든 변경 및 변형된 형태를 본 발명의 범위로 포함하여 해석하여야 한다.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. 감송향을 유기 용매로 추출한 유효성분을 포함하는 신장암 예방 및 치료용 조성물.A composition for preventing and treating kidney cancer, comprising an active ingredient extracted with sweet persimmon as an organic solvent.
  2. 제 1항에 있어서, 상기 유기 용매는 에탄올인 것을 특징으로하는 신장암 예방 및 치료용 조성물.The method of claim 1, wherein the organic solvent is a composition for preventing and treating kidney cancer, characterized in that ethanol.
  3. 제 2항에 있어서, 상기 에탄올로 추출한 유효성분은 50℃에서 24시간 동안 추출되는 것을 특징으로 하는 신장암 예방 및 치료용 조성물.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. 제 3항에 있어서, 상기 에탄올 추출물은 45℃ 감압 조건에서 건조 및 농축되는 것을 특징으로 하는 신장암 예방 및 치료용 조성물.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. 제 1항 내지 제4항 중 어느 한 항에 있어서, 상기 조성물은 약제학적으로 허용가능한 담체 또는 희석제를 포함하는 것을 특징으로 하는 신장암 예방 및 치료용 조성물.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. 제 1항 내지 제4항 중 어느 한 항에 있어서, 상기 신장암은 신세포암(kindney cell cancer)인 것을 특징으로 하는 신장암 예방 및 치료용 조성물.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. 화장품학적으로 허용 가능한 화장품 보조 첨가제를 포함하는 감송향(Nardostachyos Rhizoma) 에탄올 추출물을 유효성분으로 함유하는 신장암 예방용 화장료 조성물.A cosmetic composition for preventing kidney cancer comprising Nardostachyos Rhizoma ethanol extract as an active ingredient comprising cosmetically acceptable cosmetic auxiliary additives.
PCT/KR2012/002465 2011-03-31 2012-04-02 Composition for renal cancer treatment and composition for cosmetic containing nardostachyos rhizoma extract WO2012134250A2 (en)

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KR101440574B1 (en) * 2013-02-27 2014-09-17 안동대학교 산학협력단 Pharmaceutical composition comprising the extract of nardostachys chinenesis as an effective component for prevention or treatment of diseases related to thrombosis and health functional food comprising the same
US20190314436A1 (en) * 2016-11-04 2019-10-17 Korea Bio Medical Science Institute Co., Ltd Pharmaceutical composition for prevention or treatment of liver cancer and health functional food

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WO2004112692A2 (en) * 2003-05-19 2004-12-29 Amazon Biotech Inc. Therapeutic herbal compositions
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JP2001192338A (en) * 2000-01-11 2001-07-17 Pola Chem Ind Inc Agent for promoting recovery from adverse effect of stress and skin preparation which contain the same and is useful for external use
WO2004112692A2 (en) * 2003-05-19 2004-12-29 Amazon Biotech Inc. Therapeutic herbal compositions
US20070122495A1 (en) * 2005-11-28 2007-05-31 Sahajanand Biotech Pvt. Ltd. Herbal composition to improve psychological functions as an anxiolytic, tranquilizer, and non-narcotic sedative, as well as other physiological functions
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