KR20020084872A - Composition comprising an extract of alpinia katsumadai for hormone replacement theraphy - Google Patents

Abstract

PURPOSE: A composition containing an Alpinia katsumadai Hayata extract as an effective component is provided. The composition can be used as a pharmaceutical composition for hormone replacement therapy of a menopausal woman because it exhibits human estrogen receptor-binding activity. CONSTITUTION: The pharmaceutical composition for treatment of hormone replacement therapy contains 0.1 to 50% by weight of an Alpinia katsumadai Hayata extract and the extract is orally administered at a daily dose of 0.001 to 100mg/kg. The Alpinia katsumadai Hayata extract is prepared by extracting with water or lower alcohol selected from methanol, ethanol, normal propanol, iso-propanol, normal butanol and a mixture thereof at 60 to 100deg.C, filtering or centrifuging and then concentrating under reduced pressure.

Description

COMPOSITION COMPRISING AN EXTRACT OF ALPINIA KATSUMADAI FOR HORMONE REPLACEMENT THERAPHY}

The present invention relates to a composition for treating hormone replacement using an extract of Alpinia katsumadai as an active ingredient, specifically, it is isolated and purified using water or a lower alcohol from the scalp and is directed to a human estrogen receptor. The present invention relates to a pharmaceutical composition comprising as an active ingredient a chodochoo extract showing binding activity.

In the United States alone, more than 4000 million women enter menopause each year. The average age of women who reached the last menstrual age was about 28 years, of which about 75 to 85 percent would develop estrogen deficiency (Hammond et al., Fertil. Steril. , 37 (1)). : 5-25, 1982). One of the most common diseases following the onset of ovarian failure is 'hot flash' or vasomotor complexes, which are suddenly occurring and generally last for several minutes, frequently observed red flashes and often Accompanying dizziness, nausea, headache, palpitations, and sweating, more than 90% of these symptoms can be alleviated by providing adequate estrogen.

There are many postmenopausal symptoms caused by chronic low estrogen, the most serious of which are osteoporosis and ischemic heart disease. It has been reported that 25% of women older than 60 years have developed vertebral compression fractures as a result of osteoporosis associated with estrogen deficiency, and 50% have developed vertebral fractures at age 75. The very high rate of hip fractures in old age is also attributable to this osteoporosis.

Hormone replacement therapy (HRT) with estrogens or estrogen / gestagen combinations is now known as the standard method for treating symptoms associated with menopause (Ernster, VL et al., Prev. Med ., 17: 201-223, 1988).

Estrogens protect the cardiovascular system, bone (reduced risk of osteoporosis) and central nervous system (avoid hot flashes). However, chronic use of estrogen in hormone replacement therapy increases the risk of endometrial cancer (Ernster, VL et al., Prev. Med ., 17: 201-223, 1988). Simultaneous use of gestagen to reduce this risk inhibits the stimulating effect of estrogen on the endometrium (Gibbson, WE, Am. J. Obstet. Gynecol. , 154: 46-61, 1986), but in contrast, estrogen and In the case of the combination treatment of gestagens , there is a problem of deteriorating the protective effect of the estrogen component on plasma lipids (Lobo, R. Am. J. Obstet. Gynecol ., 166: 1997-2004, 1992).

Moreover, with estrogen / gestagen based on lower hormonal dosages than using oral contraceptives, undesirable intra-menstrual bleeding can occur (Hillard, TC et al., Am. J. Obstet. Gynecol. , 167: 1-7, 1992). Finally, recent papers show that many gestagens may increase the risk of breast cancer (Staffa, JA et al., An Epidemiologic Review , 57: 473-491, 1992; King, RJB, J. Ster. Biochem Molec. Bio ., 39: 8111-8118, 1991).

In particular, estrogen replacement therapy is a more complex issue for women with the uterus. Estrogen treatment is associated with an increased incidence of endometrial cancer due to proliferation of the endometrium induced by continuous 'competitive' estrogens. Regular administration of progestin seems to reduce the rate of endometrial cancer in postmenopausal women who receive estrogen by several folds through the antiproliferative effect (Barbieri et al., Menopause Management , July / August, 12-24, 1992). However, the combination of estrogen and progestin frequently causes undesirable uterine bleeding and has the disadvantage that the cardiovascular effect of estrogen can be minimized by progestin. Nevertheless, the estrogen regimen, plus a series of concurrent progestins, is now used as a common form of hormone replacement therapy in postmenopausal women with a uterus.

In addition, estrogens have been reported to be effective in treating dementia (Black SE, Patterson C., Feightner J., Can. J. Neurol. Sci., Feb: 28 Suppl 1: S56-66, 2001), neuroprotective action (Dubal DB., Wise PM., Endocrinology, Jan; 142 (1): 43-8, 2001) and improvement effects on neurodegenerative diseases (Kim H., Xia H., Li L., Gewin J., Biofactors, 12 (1-4): 243-50, 2000).

Meanwhile, Alpinia Katsumadai semen is a herb belonging to the family Zingiberaceae. Chodugu is distributed in southern China, Taiwan and Hong Kong, and is cultivated in tropical regions, and has the effects of detoxification, constitution improvement, oral hygiene, and anti-inflammatory agents.

Edinosa is a perennial herb, with thick and short roots, with greenish stem. When the fruit turns yellow in autumn, harvest it and dry it for 7 to 8 days in the sun, peel off the skin and dry it again in the sun. The shape of the head nut is a spherical round shape of seeds, and is 15 to 27 mm in diameter. The outer side is grayish brown, and in the middle, it is divided into three parts by the gray-white diaphragm, and the seeds are divided. Seeds are smooth and do not fall easily. Oval polyhedron, 3 to 5 mm long, about 3 mm in diameter, covered with a light brown membranous apiary, and hard. The shoes have an unusual aroma and taste spicy and bitter.

Accordingly, the present inventors have made efforts to find a natural product that can be used for estrogen replacement therapy. As a result, the present inventors have found that the cedar extract exhibits binding activity to the human estrogen receptor and thus can be effectively used for hormone replacement treatment of postmenopausal women as an estrogen-like substance. Was completed.

It is an object of the present invention to provide a hormone replacement therapeutic composition containing natural extracts as analogs of hormones such as androgens or estrogens.

1 is a result of examining the estrogen generating ability (estrogenicity) of the head nut extract using recombinant yeast expressing the human estrogen receptor.

C; No treatment group E2; 17β-estradiol

1-1; Edinary bulrush extract 1 mg / ml 1-2; 100 ㎍ / ml of Edinosa extract

In order to achieve the above object, the present invention provides a pharmaceutical composition for hormone replacement therapy comprising a head nut extract that produces an estrogen-like substance showing binding activity to the estrogen receptor.

In another aspect, the present invention provides a health food composition for hormone replacement therapy comprising the cedar extract extract that produces an estrogen-like substance showing binding activity to the estrogen receptor.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for hormone replacement therapy comprising as an active ingredient a head nut extract that produces an estrogen-like substance showing binding activity to the estrogen receptor.

The nutmeg extract contained as an active ingredient in the pharmaceutical composition of the present invention can be obtained by extracting from a nutmeg with a conventional method using water or a lower alcohol.

For example, the hot water extraction method is dried and pulverized chodugu, 1 to 20 times the water by weight ratio, and then extracted for 1 to 6 hours at 60 to 100 ℃ filtration or centrifugation and the supernatant concentrated under reduced pressure to Prepare an extract

In addition, the lower alcohol extraction method is added 5 to 20 times the lower alcohol to the cedar, refluxed for 1 to 6 hours while heating to 60 to 100 ℃, cooled and filtered and concentrated under reduced pressure using a rotary evaporator to extract the cedar extract In this case, the filtration process may be repeated two or more times with respect to the residue remaining after filtration. In this case, as the lower alcohol, methanol or methanol aqueous solution, ethanol or ethanol aqueous solution, normal propanol, iso-propanol, normal butanol and mixtures thereof may be used, and ethanol of 60 to 80% is most preferred.

In order to investigate whether the ethanol extract extracted from the cedar by the extraction method exhibits hormonal activity, the ethanol extract of the cedar is expressed in a recombinant yeast expressing a human estrogen receptor. The reaction was measured by application.

Treatment of the test substance with 1 mg / ml and 100 μg / ml showed a significant difference in comparison with 17β-estradiol (E2), which was used as a positive control. High estrogen production was observed in yeast to which the head extract was applied.

Through the above experimental results, it was confirmed that the head ethanol extract of the present invention can bind to the estrogen receptor and act as an analogue of the human estrogen hormone.

Since the same effect as the administration of estrogen can be expected by administering a pharmaceutical composition containing the choledophil extract of the present invention exhibiting such an effect, the pharmaceutical composition of the present invention can be usefully used as a preventive and therapeutic agent for osteoporosis.

The pharmaceutical composition of the present invention can be administered orally or parenterally during clinical administration and can be used in the form of general pharmaceutical preparations.

That is, the pharmaceutical composition may be administered in various oral and parenteral dosage forms in actual clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used Is prepared using. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose (Sucrose) or lactose (Lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations 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 a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The pharmaceutical compositions of the invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular.

Typical daily dosages of the turmeric extract of the present invention range from 0.001 to 100 mg / kg body weight, and may be administered once or in divided doses. However, it is to be understood that the actual dosage of the active ingredient should be determined in light of several relevant factors such as the route of administration, the age, sex and weight of the patient, and the severity of the patient, and therefore the dosage may be determined in any aspect of the invention. It does not limit the scope.

In another aspect, the present invention provides a health food composition for hormone replacement therapy comprising the cedar extract extract that produces an estrogen-like substance showing binding activity to the estrogen receptor.

Since the same effect as the administration of estrogen can be expected by ingesting the health food composition containing the extract of the chorus nutmeg of the present invention, the health food composition of the present invention can be usefully used as an agent for preventing and treating osteoporosis. In order to act as a hormone-like substance, foods to which the chondura extract of the present invention can be added include, for example, various foods, beverages, gums, teas, vitamin complexes, and health supplements.

Food preparation can be prepared by adding to the raw material in the preparation of food or mixed with the cooked food properly, in this case, the content of the extract from the final production of food or beverage is 0.1 to 20% by weight Range.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Reference Example 1 Recombinant Yeast

Saccharomyces cerevisiae ER + LYS 8127 (YER) (Donald C. McDonnell, Duck University [Chemical Industry Institute of Toxicology, USA]) used to investigate the hormonal activity of the turmeric extract of the present invention. Donated by the doctor) is a hER, hAR gene and reporter system whose expression is regulated by the CUP1 metallothionine promoter, which stably estrogen response elements and β-gal genes. Recombinant yeast comprising a vector to express. ER + LYS 8217 is a saccharomyces cerevisiae 3.35 g / ml yeast nitrogen base (yeast nitrogen base, Difco, USA), 2% dextrose (dextrose, Gibco, USA), 30 μg / ml L-lysine Incubated in growth media containing -HCl (L-lysine-HCl, Sigma, USA), 35 μg / ml L-histidine-HCl (L-histidine-HCl, Gibco, USA) It was. Recombinant yeast was stored at −80 ° C. or lower by adding 20% glycerol (glycerol, Sigma, USA) to the growth medium until it was used for hormonal activity experiments using the cranial extract.

Example 1 Preparation of Cereal Extract

2 liters of 70% ethanol was added per 100 g, and the mixture was refluxed for 3 hours while heating to 90 ° C, cooled and filtered, and concentrated under reduced pressure using a rotary evaporator, thereby obtaining a chondulum ethanol extract.

Example 2 Determination of Hormonal Activity (Estrogenicity) of Cereal Extract

In order to confirm that the ethanol extract of the scalp was isolated and purified from Example 1 shows hormonal activity, the scalp extract was extracted in ER + LYS 8127 (YER) of recombinant yeast Saccharomyces cerevisiae expressing human estrogen receptor. The estrogen production ability was investigated by application.

Specifically, the recombinant yeast stored at −80 ° C. according to Reference Example 1 was dissolved, and then placed in the same growth medium, and then grown at 30 ° C. and 300 rpm in a shaking incubator. Proliferated recombinant yeast was diluted to the appropriate concentration, 500 μM of CuSO ₄ (Sigma, USA) was added and the appropriate amount was dispensed into 50 ml cornical tubes. The tube was treated with 1 mg / ml and 100 μg / ml, respectively, as a test group, and treated with 17β-estradiol (E2, 10 ^-9 M) as a positive control, and 100% as a solvent control. Ethanol (absolute ethanol) was used.

The experimental group and the control group treated with the respective materials were incubated for 18 hours at 30 ° C. and 300 rpm in a shake incubator, and then the culture solution of each tube was diluted to the same concentration and a 96 well microtiter plate (96 well microtiter plate, Costar, USA) was dispensed into 3 wells 100 μl each group. 2 mg / ml o-nitrophenyl-β-D-galactopyranoside (o-nitrophenyl-β-D-galactopyranoside, Sigma, USA), 0.1% lauryl sulfate (Sigma, USA), 50 mM β-mercaptoethanol (β-mercaptoethanol, BDH chemical, England), and 200 U / mL Z buffer solution (oxalyticase, Enzogenetics, USA) were dispensed and mixed After 20 minutes, the color development was measured at a wavelength of 420 nm using a microplate reader.

As a result, the experimental group treated with the choledophil extract of the present invention at concentrations of 1 mg / ml and 100 µg / ml, respectively, showed a significant difference compared to the positive control treated with 17β-estradiol (E2). The positive control group treated with 10 ⁻⁹ M of 17β-estradiol (E2) showed an absorbance of about 2.7, whereas the experimental group treated with 100 g / ml and 1 mg / ml of cedar extracts had absorbances of 6.0 and 3.0, respectively. It showed relatively higher estrogen generating capacity than 17β-estradiol, which is conventionally used as a hormone replacement therapy.

Therefore, the extract of the head of the present invention is an estrogen-like substance showing estrogen receptor binding activity, and the composition including the same may be useful for the hormone replacement treatment of diseases such as osteoporosis.

As described above, since the chondola extract of the present invention includes an estrogen-like substance that binds to the estrogen receptor, it may be usefully used as a pharmaceutical composition for treating hormone replacement in postmenopausal women.

Claims (5)

A pharmaceutical composition for the treatment of hormone replacement in postmenopausal women, which contains the extract of Edinosa vulgaris. The pharmaceutical composition of claim 1, wherein the hormone is estrogen. The pharmaceutical composition according to claim 1, wherein the choledophil extract is prepared by extracting with water or lower alcohol. 4. A pharmaceutical composition according to claim 3, wherein the lower alcohol is selected from the group consisting of methanol or methanol aqueous solution, ethanol or ethanol aqueous solution, normal propanol, iso-propanol, normal butanol and mixtures thereof. Health food composition for hormone replacement treatment of postmenopausal women with the extract of the head nutmeg.