EXTRACT OF SMILAX CHINA ROOT HAVING ESTROGEN-LIKE EFFECT

The present invention refers to a root estrogen receptor α and β [cheng future composition for oestrogen by being coupled signals derived from coupling of an estrogen receptor can be activating delivery it will be small, in addition to a new approach based on forced capable of promoting proliferation of breast that result are disclosed.

Estrogen receptor (estrogen receptor) if estrogen is estrogen binding other gene involved in signal transduction for regulating transcription in conjunction with a transcription regulatory factor could be bonded each other. The human estrogen deficient in this signal the delivery work smoothly and neutralise, utilization is equal to various diseases occur.

The 45 - 55 cells of estrogen deficiency such electrical signals related conditions include nothing border crossing before adult females mainly, more or more, qualitative xeransis, fluoxetine, fatigue, excited, lowering concentration, insomnia, depression, memory upper actual proof, neural hypersensitivity, treating timidity, arthralgia, obesity, osteoporosis, wine, hyperlipidemia, hypertension, stroke, skin diseases, herbal inhabit orgin diseases, is nitro, uterine cancer, breast cancer, dry hair and skin like known.

In the present invention a root estrogen receptor α and β [cheng future composition for delivery it will be small by being coupled thereby activating signal using human ER reporter assay kit was certification that the ferroelectric layer. The kits are estrogen receptor α or β which express these receptors to engage the marker gene (such as luciferase) is expressed estrogen such as material designed to be used for a cells. And promoting proliferation of estrogen are expressed that was also certification of a woman's breast. These results indicate that [cheng future composition for estrogen such as action which indicates that a root, the root extract of said through the administration of estrogen deficiency such as diseases caused [cheng future composition for the prevention, treatment or amelioration can be can occur no matter what.

The party industry plant extraction method of the present invention [cheng future root extract composition for using conventional solvents and the number bath 1308. The solvent water, alcohol 1 - 4 carbon atoms, acetone, hexane, ethyl acetate and carbon dioxide selected from one or more can be mixed with a solvent, preferably water, ethanol or a combination of solvent and slipperiness, more preferably 20 - 50% ethanol in affinity with a mixed solvent (v/v) using a good ancestry. The amount of solvent based on the weight of the growing root [cheng future composition for 0. 1 - 100 Times the volume amount (v/w) can be, preferably 0. 5 - 50 Times the volume amount now. To extraction temperature can be 0 - 100 °C, preferably 20 - 60 °C maintained good, can be extraction time is 1 hr or longer, preferably 12 - 48 time now. In this way said action effect can be extract number such as estrogen such as high pressure liquid coolant.

The material of the present invention extract number [cheng future composition for a bath (Smilax chinaL.) or a number can be used to commercially available roots grow grudge without, a dried, drying but wasn't grudge without number can be used, preferably using a toe luck zero therefore sold to seller.

In the present invention [cheng future composition for the root extract of pharmaceuticals, food, cosmetic, such as can be use in medicine external width.

The, root extract of the present invention according to (KFDA) conventional about number [cheng future composition for medicinal food into government office number number number number based on the number of health supplementary food reference or use to elder brother anger elder brother anger enhancement type can be disclosed.

[Cheng future root extract composition for the conventional method, administration method, dosage forms and the dermatologically acceptable carrier and therapeutic purposes about number are mixed together to dilution or, in that the carrier in the form of sealed container to elder brother anger number can be disclosed.

Said carrier is used as a dilution number brine, buffer number, dextrose, water, glycerol, ringer's solution, peak lactoperoxidase, sucrose, calcium silicate, methyl cellulose and ethanol from at least 1 selected from the group consisting at least one oral administration for parenteral administration the carrier powder, granules, cap, syrup, solution number, positive number, suppository, lung coil (pessaries), ointment, cream or aerosol such as number number can be-high pressure liquid coolant. Only, and not the confined within said carrier carrier of the present invention. The, in addition to the colorectal cancer oral parenteral administration, intravenous, peritoneal, muscle, artery, transdermal, nasal, administration of big active suction or the like.

Filling said number type number, number clause cohesion, lubricating number, wet number, perfume, emulsion number, such as antiseptic number further comprises a mammalian administered of the active ingredient after rapid, sustained or delayed release can be elder brother anger [...] number to number. The dose of the present invention to a patient's state, routes and dosage form of the present invention is not limited to such symptoms can be adjustable and in the field for use in a wide variety if the person with skill in the art nontrivial range but, in the present invention 1 to 100 mg per day per 1 kg body weight in an amount effective to empirical typically a continuous or intermittent administration thereof can be pathogenic substrate.

[Cheng future composition for an effective amount of a root extract of said reference, root extract of the present invention refers to a cosmetically acceptable carrier [cheng future composition for cosmetic itself or a cosmetic composition which number [...] mixing, said skin cosmetic composition, plant, encapsulated into, massage cream, essence and nutritional pack cosmetic form can be use and number of bath.

In addition, root extract composition for an effective amount of said [cheng future reference, the present invention refers to a food containing low number [cheng future root extract composition for cosmetically acceptable carrier including food itself or to [...], meat workpiece said food composition, article number fish, bean curd, powder, dead, Chow mein if such as noodles, soy sauce, soybean paste, red pepper paste, such as seasoning food mixing field, source, confectionery, such as milk or cheese type manufactured goods, such as pickles or kimchi pickling, food, fruit, vegetables, soy milk, fermented beverage drink such as food including an can be used. A cosmetically acceptable food carrier can be about number be used cosmetically acceptable carrier.

[Cheng future composition for an effective amount of said root extract of reference, the present invention refers to [cheng future composition for cosmetically acceptable carrier composition including about itself or a root extract of number to number [...] medicine external width, said medicine external width clean number disinfecting composition, bath number, green, wet tissue, cell number soap, hand wash, mask, bath can be use and number of medicine external width number ointment form.

Hereinafter, the present invention through further detailed in the embodiment to less than 1000. In the embodiment for the present invention is to exemplify these only since, in the embodiment of the present invention these films in the range number by one not be interpreted.

In the embodiment 1. [Cheng future number of root extract composition for bath

Voice recorder galenical, i.e. [cheng future composition for a seller therefore rolled roots from 0. 5 - 2. 5 Mm was crushed. For the extraction of a predetermined galenical 0. 5 To 50 times the volume amount of 20 - 50% ethanol solution (solution mixing with ethanol) (v/w) into water, at a temperature of 20 - 60 °C stirring time extracted from the 12 - 48. Filtering the obtained extract of filter directly 8 micro m hereinafter, cream or lyophilized extract after the last obtained.

Experiment example 1. [Cheng future composition for estrogen receptor α and β binding activity of verification of root extract

Root extract composition for [cheng future possibility estrogen replacement of function key, estrogen receptor α and β his experiments with binding activity.

Number tested in human ER reporter assay kit (Indigo Bioscience, USA) high pressure liquid coolant in said in the embodiment 1 extract using estrogen receptor α and β binding activity were measured.

Estrogen receptor α β expressed reporter cells on each recovery medium selected from 1:1 96 a-well tissue culture plate (96 a-well tissue culture plate) to 1 × 10⁴ Unit 100 by dividing his micro l cells/well. Then, DMSO (dimethyl sulfoxide) at a concentration of 100 mg/Ml to extract of said in the embodiment 1 according to the next cells cultured in 24 hours was 37 °C dissolving concentration. Voice controls are equal to extract the deflection from the distilled water collecting volume, positive controls are estrogen (17 a-estradiol) 6 a. 4PM at night. The supernatant culture is terminated plate number was a stand-alone. Remaining cells in each well group 5 minutes after adding 100 micro l volume luciferase detection reagent luminometer using light emission amount is left for a long time. Estrogen receptor binding activity of verification is processed voice controls of distilled water 1 is reduced terms of the bill.

Estrogen receptor α measured for binding activity, estrogen to 6. 4PM positive controls are treated with regulated contrast 2. 73 Times increased as in the picomolar. In the embodiment 1 1 100 micro g/Ml micro g/Ml electrically coupled to the active extract of processing group from a concentration dependent increase in the 24.4, among 100 micro g/Ml 1 contrast controls at a predetermined concentration. 73 Times increased binding activity as in the picomolar (also 1A reference).

Estrogen receptor β measured for binding activity, estrogen to 6. 4PM positive controls are treated with regulated contrast 2. 93 Times increased as in the picomolar. In the embodiment 1 1 100 micro g/Ml micro g/Ml electrically coupled to the active extract of processing group from a concentration dependent increase in the 24.4, among 100 micro g/Ml 1 contrast controls at a predetermined concentration. 43 Times increased binding activity as in the picomolar (also 1B reference).

On the experiment result classifies, [cheng future composition for estrogen receptor α and β engage both on root extract of the present invention, one having a higher affinity than to estrogen differently from estrogen receptor α can be know.

Experiment example 2. [Cheng future root extract composition for breast cell proliferation effect verification

[Cheng future composition for root extract [...] similarly estrogen receptor is expressed estrogen functional groups to be verification whether his promotes mammary cells.

2 - 1. Cell culture

Estrogen receptor is expressed estrogen receptor is expressed breast cells MCF-a 7 and not of a woman's breast MDA-a MB231 cells was used. Medium number of estrogen activity demonstrating component times for, 10% charcoal treated FBS, 0. 01 Mg/Ml bovine insulin, antibiotic number (100unit/Ml penicillin, 100 micro g/Ml streptomycin) RPMI 1640 and adhesive by adding a transparent conductive layer, as well as exchanging each involve passaging cells to each medium 3 - 4 7.

2 - 2. Mammary cell proliferation effects by estrogen like activity

Induction of apoptosis caused by proliferation effect on estrogenic activity such as modifications to the method applied to the determination of the Soto. 2 × 10 96 a-well tissue culture plate⁴ Breast cells 24 hours him as unit cells/well. Then, test medium (5% charcoal dextran treated FBS and, do not contain a phenol-a red RPMI 1640) after replacing, DMSO (dimethyl sulfoxide) extract of said in the embodiment 1 according to the next cells 24 hours dissolving concentration concentration 100 mg/Ml to him as. Controls are equal to extract the deflection from the distilled water collecting volume, as well as a positive controls are in a concentration of 10 ng/Ml estrogen. MTT proliferation of whether reagent are injected into a culture is terminated plate were measured. Proliferation effect on breast cells showed 100% survival rate terms of distilled water verification is processing the bill.

The estrogen receptor is expressed cells MCF-a 7 measured proliferative effect, a concentration of 10 ng/Ml positive controls are treated with estrogen 140 breast proliferation of capacity. 0% To 40% contrast increased regulated up. Group 1 - 100 micro g/Ml concentration dependent cell proliferation has been effective concentration in extract of processing section in the embodiment 1, 50 and 100 micro g/Ml micro g/Ml concentration of cell proliferation in each rate 118. 03%, 121. 5% 18% To 21% cell proliferation-promoting effect on regulated contrast her (also 2A reference).

MDA-a MB231 cells in which the estrogen receptor is expressed not positive controls proliferation of cells of the 24.4 is insignificantly, concentration of 1 - 100 micro g/Ml extract of processing delay time in the embodiment 1 does not require such a significant cytotoxicity and toxicity period (reference also 2B) his active cell proliferation.

When on data by the experiment result, functional groups of the present invention [cheng future which will bind to the estrogen receptor α and β similarly [...] root extract composition for estrogen, promote the proliferation of breast through can be know that the ue has mechanisms of action.

Said ferroelectric layer of the present invention composition such as estrogen receptor is expressed in cells that estrogen in place can be serve as female hormone replacement number, ameliorating female hormonal imbalance machine dysfunction due to degradation, for preventing or treating may be material of has been confirmed.