KR20020084877A - Composition comprising an extract of astragalus membranaceus for hormone replacement theraphy - Google Patents

Abstract

PURPOSE: A composition containing an Astragalus membranaceus B. extract as an effective component which is estrogen-like material and effective in hormone activity is provided. Therefore, the composition is useful as a prophylactic and therapeutic agent for treatment of osteoporosis or prostate cancer as well as a health food. CONSTITUTION: The pharmaceutical composition contains 0.1 to 50% by weight of an Astragalus membranaceus Bunge extract extracted with water or low alcohol. The extract is administered at a daily dose of 0.001 to 100mg/kg through an oral, transdermal, subcutaneous, intravenous or muscular pathway. The composition contains a generally used diluent or excipient such as a filler, bulking agent, binding agent, wetting agent, disintegrating agent and surfactant, together with the extract.

Description

COMPOSITION COMPRISING AN EXTRACT OF ASTRAGALUS MEMBRANACEUS FOR HORMONE REPLACEMENT THERAPHY}

The present invention relates to a composition for treating hormone replacement using the extract of Astragalus membranaceus as an active ingredient, specifically, it is isolated and purified from water by using water or a lower alcohol and binds to human estrogen receptor. It relates to the Astragalus extract and a pharmaceutical composition having the same as an active ingredient.

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, estrogen's regimen, plus a series of concurrent progestins, is now used as a common form of hormone replacement therapy in postmenopausal women.

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).

On the other hand, Astragalus membranaceus is a herb belonging to the legume (Leguminosae), according to the commentary of Ishjin of the New Farm, the color is yellow and is named in the sense of protecting the human body for a long time. Since ancient times, Astragalus root has been used a lot as a medicine after ginseng in the private sector, and it has been used variously as a tonic, diuretic, and antiperspirant. In particular, in oriental medicine, Hwanggi is used for hundreds of prescriptions such as Bogiikgitang, Hwanggigunjungtang, Jeopjeondaebotang, Kamidaebotang, Palbohoechuntang, and Cheongseoikgitang.

Astragalus is distributed not only in Korea, but also in China, Inner Mongolia, Japan (Hara; Pallas Subsp.Kita; Hands-Mazz.), Egypt, and Bulgaria.

Until now, the triterpenoid glucoside (triterpenoidglycoside) line and the isoflavonoid line among flavonoids have been reported mainly as components. Specifically, polysaccharides isolated from Astragalus have been reported to improve the function of T cells in cancer patients, and triterpenoids and flavonoids are known to have antioxidant, anti-inflammatory and blood pressure lowering activities (Huaping L., Yau, ZO and Bo, G., 1994, 3 (102), 138-141; Geng, CS, 1986, 6 (1), 62-64); Zhang, Y. D. et al., 1992, 27 (6), 401-406; Harborne, J. B. and Baxter, H., 1993, Taylor & Francis, London, Washington DC., Pp 673; Tang, W. and Eisenbrand, G., 1992, Springer-Verlag, Berlin, Heidelberg, New York, pp. 191).

Accordingly, the present inventors have made efforts to find a natural product that can be used for the treatment of estrogen replacement. As a result, the present invention revealed that the extract of Astragalus is an estrogen-like substance showing binding activity to the human estrogen receptor, and thus can be effectively used for hormone replacement treatment in postmenopausal women. Was completed.

It is an object of the present invention to provide a hormone replacement therapeutic composition containing a natural extract as an active ingredient as a hormone analogous substance such as androgen or estrogen.

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

C; No treatment group E2; 17β-estradiol

1-1; 100 g / ml 1-2 of Astragalus extract; Astragalus Extract 1mg / ml

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

In another aspect, the present invention provides a health food composition for hormone replacement therapy using the Astragalus extract to produce an estrogen-like substance showing a 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 the extract of Astragalus to produce an estrogen-like substance showing binding activity to the estrogen receptor.

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

For example, the hot water extraction method is dried and pulverized sulfuric acid, added 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 extract the sulfuric acid Manufactures

In addition, the lower alcohol extraction method is added to 5 to 20 times lower alcohol in sulfuric acid, 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 obtain a sulfuric acid 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 Astragalus by the extraction method shows hormonal activity, the ethanol extract of Astragalus is applied to a recombinant yeast expressing a human estrogen receptor. The reaction was measured.

As a result of treatment of test substance with 1 mg / ml and 100 ㎍ / ml, there was a significant difference in comparison with 17β-estradiol (E2) used as a positive control. High estrogenicity was observed in yeast to which the extract was applied.

Through the above experimental results, it was confirmed that the ethanol extract of the present invention binds to the estrogen receptor and can 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 comprising the Astragalus 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 drug replacement therapy.

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. Solids for oral administration include tablets, pills, powders, granules, capsules, and the like. Such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, and the like. (Sucrose) or lactose (Lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium styrate 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 replacement treatments, 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 Astragalus 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 way of the invention. It does not limit the scope.

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

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

Food products can be prepared by adding the Astragalus extract of the present invention to a raw material during food preparation or by appropriately mixing the cooked food, in which case the content of Astragalus extract in the finally prepared food or beverage ranges from 0.1 to 50% by weight. .

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

Dr. Donald C. McDonnell of Saccharomyces cerevisiae ER + LYS 8127 (YER) (Duck University [Chemical Industry Institute of Toxicology, USA]) used to investigate the hormonal activity of the Astragalus extract of the present invention. Donor) is a hER, hAR gene and reporter system whose expression is regulated by the CUP1 metallothionine promoter, which stably expresses the estrogen response element and β-gal gene. It is a recombinant yeast containing a vector. 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 used for hormonal activity experiments with Astragalus extract.

Example 1 Preparation of Astragalus Extract

A sulfuric acid was added 2 L of 70% ethanol per 100 g and refluxed for 3 hours while heating to 90 ℃, cooled and filtered and concentrated under reduced pressure using a rotary evaporator to obtain a sulfuric ethanol extract.

Example 2 Measurement of Hormone Activity of Astragalus Extract

In order to confirm that the ethanol extract of Astragalus isolated and purified from Example 1 exhibited hormonal activity, the Astragalus extract was applied to a recombinant yeast Sakaromyces cerevisiae expressing human estrogen receptor ER + LYS 8127 (YER). The estrogen production ability was investigated.

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 experimental group was treated with 1 mg / ml and 100 μg / ml of the Astragalus extract of the present invention as an experimental group, and 17β-estradiol (E2, 10 ^-9 M) was treated as a positive control, and 100% ethanol as a solvent control. (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 three wells of 100 μl in 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 Astragalus 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 absorbance of about 2.7, whereas the experimental group treated with 100 g / ml and 1 mg / ml of Astragalus extract showed absorbances of 1.6 and 3.5, respectively. It showed relatively higher estrogen-generating ability than 17β-estradiol, which is conventionally used for hormone replacement therapy.

Therefore, the Astragalus extract of the present invention is an estrogen-like substance showing estrogen receptor binding activity, the composition comprising the same can be usefully used for the hormone replacement treatment of diseases such as osteoporosis.

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

Claims (5)

A pharmaceutical composition for treating hormone replacement in postmenopausal women using the extract as an active ingredient. The pharmaceutical composition of claim 1, wherein the hormone is estrogen. The pharmaceutical composition according to claim 1, wherein the Astragalus 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 using the extract as an active ingredient.