Soy extract, method of its preparation and pharmaceutical composition

 

(57) Abstract:

The invention relates to the field of medicine. Soy extract contains from 0.6 to 1.5 weight. including saponins soy group per 1 weight.h. glucosidic of isoflavones contained at least in the amount of 13% in the extract. The extraction is carried out for low-fat soy beans or flour aliphatic alcohols or their aqueous mixtures in the reciprocal ratio of 3:2 to 2:3. The extract was concentrated, purified. Extracted aliphatic hydrocarbons, not miscible with water, alcohols, is not miscible with water. The extract was concentrated and dried. The pharmaceutical composition contains soy extract for the treatment and prevention premenopausal and postmenopausal symptoms, alcoholism, treatment for cancer of the breast, prostate. The invention improves the efficiency of funds. 3 S. and 8 C.p. f-crystals, 1 Il., 3 table.

The invention relates to new extracts obtained by extracting Mature whole soy beans or oil-free soya flour Glycine max (L.) Merril: family - Leguminosae, their preparation and compositions containing these extracts. New extracts are characterized by the content of isoflavones and saponins at a certain ratio.

Contained in soy saponins can be divided into three classes based on the chemical structure of their triterpene components saponins soy groups a, b and E (K. Okubo et al., 1994, ACS Symp., Ser. 546, 330).

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Isoflavone components include glucoside isoflavones (daidzin, genistin and glycitin), which may include acyl radicals, for example monilinia radicals, United sacharides chain.

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In accordance with the biomedical literature published in recent years, epidemiological information, especially in relation to the population of the countries of the East, largely using culinary products are soy-based, consumption of such foods significantly reduces premenopausal and postmenopausal symptoms in women (A. Cassidy, 1996, RTOs. Nutrition Soc. , 55, 339-417). These effects, for which there is no sufficiently precise scientific basis, usually associated with genistein, daidzein and glycitein, which isoflavonoid the aglycones, which are present in different food products, prepared with IP the studies in vitro have shown, that these compounds act in accordance with the mechanism ensuring competition estrogen mammals, despite the fact that their activity is estimated at 500 to 1,000 times lower than that of estradiol (D. A. Shutt & P. I. Mor, 1972, J. Endocrinol, 52, 299-310).

Further, in accordance with the biomedical literature and recently published epidemiological information specific to populations of the East, where increasingly adopted the use of soy in food, consumption of such foods is associated with a significant reduction in the incidence of breast cancer in women and prostate in men (A. Nomura, B. E. Henderson, J. Lee, 1978, Amer. J Clin. Nutrition, 31, 2020-2025; T. Hirayama, 1986, In "Diet, Nutrition &Cancer, 41-53, eds. Y. Hayashi, M. Nagao, T. Sugimura, S. Takayama, L. Tomatis, L. M. Wattenberg & G. N. Wogan, Jap. Sci. Soc. Press, Tokyo; R. K. Severson, A. M. Y. Nomura, J. S. Grove, G. N. Stem-Ethel, 1989, Cancer Res., 49, 1857-1860). Also these facts, which have not yet received a reasonable scientific explanation, is usually associated with genistein, daidzein and glycitein, which isoflavonoid the aglycones, which are contained in various food products are soy-based.

These isoflavones have been studied in models in vitro to determine their ability to interact with protein kinases, in particular with the receptor, i.e., enzymes, ViDi the numerous attempts to obtain medicines based on soy extracts aimed at the prevention premenopausal and postmenopausal symptoms, as well as intended for the prevention and treatment of cancer. Some issued patents and patent applications describe the composition of new soy extracts, obtained by chemical or enzymatic hydrolysis glucosidic of isoflavones contained in soy beans or soybean seedlings (Japanese application Kikkoman Corp. - J-08291191, J-07173148, J-0511707566; application for international patent G. E. Kelly, WO 93/23069). All of these publications deal exclusively with the preparation of isoflavones in high concentrations and their activities in relation to the control of premenopausal and postmenopausal disorders and their antitumor activities.

To date, found that in contrast to what was reported earlier extracts, which contain glucoside isoflavones and soy saponins group In a certain ratio, are much more active in comparison with "pure" isoflavones in the prevention and treatment of premenopausal and postmenopausal symptoms and prevent or treat cancer.

Another aspect of the present alcoholno addiction. All these phenomena can be United by the common term "alcoholism": they all pose a serious challenge for the whole of modern society (G. L. Gessa, 1994, "Bisogno compulsive di bere e principio del piacere" ["The compulsion to drink and the pleasure principle"], Med. tossicodipend. ["Med. Dru Dependence], II, 5). In Italy, for example, more than 9% of the total population (about 5 million people) are "deep drunkards, and more than 1 million people suffer from alcohol dependence (F. P. Calamo-Spechhia, 1991, "Epidemiologia Dell alcolismo in Italia" ["Epidemiology of alcoholism in Italy"], Atti VII Congr. Naz. S. I. A. [Repts 7th Natl. Congr. S. I. A.], Mediserve, Rome, 295-301). These figures increase when calculating accept such countries as the USA, where more than 13 million people are characterized by alcohol dependence. Addiction to alcohol and present alcohol dependence lead to very significant socio-economic losses (since 1991 in the United States such losses amounted to 200 billion dollars per year) and cause serious social and psychological problems in those who are prone to addictions.

Existing attempts to treat alcoholism, complementary psychological methods ("group therapy" and so forth) associated with the use of drugs such as disulfiram and urea calcium, which affect the metabolism of alcohols with the camping inductor gag reflex, and also all other unwanted phenomena accompanying the use of alcohol.

In accordance with the previous prior art only plants used in the treatment of alcoholism, are Pueraria Pueraria lobata (product of the roots of Radix puerarie) and sage Salvia multiorrhiza, which are widespread in traditional Chinese medicine and was the object representation in patent applications WO 93/00896 and WO 96/35441. In addition to the use of these extracts in these patent applications describe the use of pure compounds, such as daidzein and their semi-synthetic derivatives (WO 93/00896), or diterpenoids, such as tension and milliron (WO 96/35441). The impact on alcoholdehydrogenase with the manifestation of the above side effects was established for isoflavonoid derivatives, although this is the mechanism for diterpenoid compounds has been eliminated. Moreover, in patent application WO 96/36332 described the action of Forskolin in reducing alcohol consumption.

Unexpectedly, it was found that the extracts containing glucoside isoflavones and saponins in soybean group in a certain ratio, can be successfully used to reduce the intentional use of alcohol. These extracts by okazaniem, which is different from the one that is associated with the suppression of the activity of alcohol dehydrogenase, because the alcohol content in the plasma remains unchanged.

In addition to the mentioned applications in international patent application WO 96/10341 described food or Wellness products, including well-peeled hypocotyl of soybean seeds. If this is not the procedure of extraction and not defined ratio of isoflavones and saponins, which is addressed by the present invention.

In U.S. patent 4428876 presents a method of separation of saponins and flavonoids of leguminous plants. Described in this patent extraction of soybean using a 0.4% aqueous sodium hydroxide solution leads ultimately to the receipt of such extract, which is different from what you receive in accordance with this invention. Again was not done definition the ratio of saponins and isoflavones made in the present invention.

In Japanese patent 59088064 presents the allocation and use only the saponins. The same applies to the German patent 34-00-258. Similarly, Japanese patent 61036225 relates to the isolation and purification of saponins, and Japanese patent 62005917 - preparation of purified saponin is xtracta, characterized by a high ratio of "saponin/isoflavone, which is an anti-inflammatory drug.

In European patent application A-426998 describes the obtaining of isoflavones from soy, in particular of genistin and daijingu-Lonate. While there is no description of the extraction of saponins and determine correlations between isoflavones and saponins.

In Japanese patent 63245648 described receiving soy food product that does not contain saponins and isoflavones, which are considered as the hard components that make this type of food inedible.

Article M. Messina et al., 1991, (J. Natl. Cancer Inst., v.83, No.8, 541-546) describes the role of soy products in reducing risk of cancer, which has already been reported in the scientific literature. However, neither this article, nor in General in the literature were not reported soy extract containing saponins and isoflavones in the ratio defined in the present invention: in these works only considered pharmacological manifestations observed when using that particular extract.

In the present invention, respectively presents the extract, which contains 0.6 to 1.5 weight.h., preferably 1 weight.h. saponins soy group, one of the weights which should be at least 13%.

As shown methods HPLC-MS saponins soy group B, which have a positive effect on the activity of isoflavonoid components are characterized by the following structure:

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Further, the present invention relates to a method of obtaining the above extract, which differs in that it includes the following stages:

a) extraction of Mature whole soya beans or oil-free soya flour using aliphatic alcohols or mixtures of these alcohols with water;

b) concentrating the extract obtained in stage (a);

c) purification of the concentrated extract phase (b) from oily and lipophilic components by processing aliphatic hydrocarbons;

d) extracting the active components are not miscible with water, aliphatic alcohols;

e) concentrating the extract obtained in stage (d), and its drying.

In particular, the extracts according to the invention can be obtained by extraction of raw, whole soya beans or oil-free soya flour containing saponins soy group and glucoside isoflavones in reciprocal ratio of 3: 2 to 2:3 with the use of aliphatic alcohols themselves or their mixtures in the areas from oily and lipophilic components by processing aliphatic hydrocarbons (for example, n-hexane or n-heptane) active ingredients extracted with not miscible with water, aliphatic alcohols, such as n-butanol, Isobutanol and isoamyl alcohol. After concentration to a lesser extent the organic phase is dried under reduced pressure. Further, the present invention represents a modification of the above-described method, in which after steps (b) or (C) the concentrated alcohol extract is subjected to stage (d'), followed by stage (e):

d') adsorption of active ingredients on polistirolovoy adsorption column, jet washing the column with water and elution of active ingredients ethanol.

In accordance with this modification of typical extract according to the invention can be obtained by adsorption of active ingredients (isoflavones and saponins), which are present in the concentrated alcoholic extract of plant material, using polistirolovoy adsorption resin, such as duality resin or any other resin XAD, in particular resin XAD1180 due to its weak acidity, and then the elution of a mixture of isoflavones and saponins soy group with ethyl alcohol after thoroughly washing the column with water jet REIT from 13% to 17% (by weight) of isoflavones and 0.6 - 1.5 parts (by weight) of soybean saponins groups, depending on the quality used plant material per 1 weight. including glucosidic of isoflavones. This extract also contains significant amounts of polyphenolic substances for which confirms the materiality in connection with the manifestation of the properties characteristic of the extract.

A variant of the above method and its modifications includes the following stages:

f) the suspension of the extract obtained in stage (e) in the mixture is miscible with water, alcohol and water, and the dilution of its water-immiscible aprotic solvent;

g) heating the mixture obtained in stage (f) providing for dissolution, followed by leaving at room temperature;

h) selection precipitiously saponins soy group filtering;

i) separating the organic phase from the aqueous phase and the concentration of the organic phase with subsequent drying her with getting isoflavonoid components; and

j) mixture of saponins obtained in stage (h), and isoflavones obtained in stage (i), to obtain a final extract.

Accordingly, the extracts according to the invention, even those, which is asianshemale above ratio can be preferably obtained from extracts, obtained in accordance with the above-described method or its modification. For this purpose, the procedure can be performed in the following way: the above extract is suspended in miscible with water, alcohol such as ethanol or methanol, with a water content of 10 to 50% by volume, and diluted in a water-immiscible solvent, such as methylene chloride or ethyl acetate. The obtained heterogeneous mixture is heated to ensure complete dissolution of the extract and leave at room temperature, resulting in the precipitation of soybean saponins group Century Saponins, purity exceeding 90%, are selected by filtering. Isoflavone components, characterized by purity over 80% receive water from the mother solution by separation of the organic phase and evaporation and drying. Then saponins and isoflavones can be mixed in order to obtain an extract which is characterized by the highest content glucosidic of isoflavones and is characterized by a content of saponins soy group at the level of 0.6 to 1.5 parts (by weight) per 1 weight.h. glucosidic of isoflavones.

The preferred conditions of the individual stages of the method according to the invention ablauts Auda kilograms (kg).

Stage (a). The plant material is preferably extracted 12-17 volumes of solvent per 1 weight.h. the biomass. The temperature of extraction is advisable to maintain the level above 55oC. Each extraction of appropriate conduct for at least 4 hours In addition to ethanol suitable solvents are, inter alia, methanol, propanol and isopropanol. These solvents may contain up to 10% water.

Stage (b). It is advisable to concentration of the extract at a temperature below the 50oWith under reduced pressure. The extract is appropriate to concentrate to the alcohol content of 65 - 75%.

Stage (C). Cleaning it is advisable to use 0.3 to 0.6 volumes of aliphatic hydrocarbons per 1 part (by weight) of plant material. A suitable procedure is the one which provides the extraction of oily and lipophilic compounds.

Stage (d). Active compounds suitable to be extracted with the use of 0.2-0.4 volumes of alcohol water-immiscible solvent for 1 extraction at the rate of 1 weight.h. plant material; it is preferable to provide three extraction.

Slowiak reduced pressure.

Stage (f). The extract obtained in stage (e), be suspended in 5-10 volumes (per 1 part of extract) water-soluble alcohol using a volume ratio of alcohol and water 2:8 to 3:7. Aprotic, water-immiscible solvent should be used in an amount of from 2 to 5 volumes per 1 weight.h. extract stage (e).

Stage (g). To achieve complete dissolution of the mixture, it is advisable to heat and to withstand the conditions of heating under reflux. The mixture is then advisable to stand at room temperature for 15-24 hours

Stage (i). The organic phase is appropriate to concentrate by evaporation at a temperature below 30oWith under reduced pressure.

Stage (j). Preferably saponins obtained in stage (h), and glucoside isoflavones obtained in stage (g), is used to prepare an alcohol solution, which contains saponins and glucoside isoflavones in the ratio of 1:1, while this solution is then feasible to subject the concentration before drying at a temperature below the 50oWith under reduced pressure.

The amount of isoflavones and saponins soy group In opredelyaem in itself (And) water (CF3COOH-0,01%), (C) acetonitrile (CF3COOH - 0,01%) and (C) methanol (CF3COOH-0,01%). Individual components can be identified and characterized using mass spectrometry in combination with HPLC through thermocapillary boundary of the section.

The extracts according to the invention differ from previously known extracts their specific impact.

Against climacteric disorders "tides" of blood, insomnia and depression are the most common symptoms of menopause suffered by women in this period of life. They are accompanied by a reduction or cessation of ovarian and as a result, the decreased production of estrogen and increased luteinizing hormone (LH; follicle stimulating hormone (FSH).

In a recent study (E. M. Duker et al., 1991, Planta Med., 57, 420) reported that in the oophorectomy (removal of ovaries) in female rats observed relationship between a random increase in the level of LH and temperature changes of the skin. This relationship between the level of LH and "tides", which is observed not only in being a model rats, but also women, confirms that the dose of LH secreted can be considered as info.

Table 1 shows the results that were obtained when subjected oophorectomy in female rats was influenced by two separate fractions (isoflavones and saponins in soy group) and the extracts according to the invention.

Female rats were subjected oophorectomy using known techniques. After 15 days after surgery, animals were exposed to the effect of the tested drugs by once-daily oral administration within 15 days. Animals were killed 3 hours after the last injection of the drug. The blood was centrifuged immediately after that, and the resulting serum was stored at -25oWith until the determination of the content of LH radioimmunoassay method, in accordance with the method described NIS-wandera et al. (Niswender et al., 1986, Proc. Soc. Exp. Biol. Med., 128, 807).

As you can see, the introduction of the extracts according to the invention resulted in a statistically significant decrease in the level of LH, which was more significant compared with that determined by the sum of the individual components (synergistic effect).

The extracts used in the repeated impacts on healthy animals did not cause any macroscopic or microscopica skeleton, that confirms their estrogenic activity.

When the extracts according to the invention were administered to women during menopause (regardless of whether it happens naturally or as a result of surgical intervention), they led to changes in the content of LH in plasma and reduced menopausal disorders, such as hot flashes or depression and so on, within a few days of exposure, and with the same exposure over a long period was also marked decrease in the level of demineralization of the bones.

The extracts according to the invention also had a distinct antiproliferative activity. Table 2 shows antiproliferative activity against cell lines ovarian cancer (OVCA 433).

Cells were cultured in monolayer culture using minimal culture medium with the addition of calf serum and 200 units/ml of penicillin to maintain the sterility of the environment. For reproducibility testing, cells were treated with trypsin each week, were sown at a density of 80 thousand cells per 1 ml and incubated at 37oC in an atmosphere of air containing 5% CO2and the presence of humidity. For tested is 100 thousand cells in 1 ml and with a minimum number of substrate. After 24 h, the substrate was replaced by a fresh portion of the substrate was added for testing compounds dissolved in absolute ethyl alcohol. Control options are processed in exactly the same way, except that intended for test compounds were absent. The above-described processing is repeated with an interval of a day during the entire testing period, amounting to 72 hours, the Level of suppression of cell proliferation was assessed by direct counting their number: the growth control variants was compared with the growth of test cases, subjected to "treatment". It was found that the extracts according to the invention showed antiproliferative activity, which was greater than the sum of antiproliferative activities of its components (activity, by type of synergy). Compounds according to the invention inhibited cell proliferation in vivo: it was confirmed by measurements of the size of tumors transplanted into Nude mice lines "naked", which was carried out by usual methods described in the literature. Introduction this animal extract in doses of from 10 to 500 mg/kg resulted in a distinct de

As for the inhibitory effect on alcohol consumption, this effect was investigated using consume alcohol in rats, representing specialized line "Sardinian, preferring alcohol" (Sp) (F. Fadda, E. Mosca, G. Colombo, G. L. Gessa, 1990, "Alcohol preferring rats: Genetic sensitivity to alcohol-induced stimulation of dopamine metabolism", Physiol. Behav. , 47, 727). These animals by providing them with free choice between ethanol and water consume 6-7 grams of alcohol per 1 kg of body weight per day (despite the fact that alcohol is diluted with water three times), have been recently successfully used to determine the effect of various compounds on voluntary alcohol consumption: see, for example, A. Balakleevsky, G. Colombo, F. Fadda, G. L. Gessa, 1990, "Ro 19-4603, a benzo-diazepine receptor inverse agonist, attenuates voluntary ethanol consumption in rats selectively bred for high ethanol preference", Alcohol, 25, 449-452; F. Fadda, C. Garau, G. Colombo, G. L. Gessa, 1992, "Isradipine and other calcium channel antagonists attenuate ethanol consumption in ethanol-preferring rats". In "Alcoholism: Clinical & Experimental Research, 16, 449-452.

The animals were kept in normal conditions, could freely choose between water (water is always present) and alcohol (10% volume solution in water), which was offered only for 4 hours a day (i.e. in the first four dark hours when the usual daily mode swtich ad libitum (ad libitum). Once stabilization was achieved ratio of consumed water and alcohol, the extract at the dose of 1000 mg per 1 kg (suspendirovanie in water) was administered orally in his voluminous amount of 2 mg/kg / day for 7 consecutive days. As control was used identical volume of filler. At the end of processing the alcohol was detected up until they achieved no indicators that were installed prior to the test.

Table 3 shows the effect of repeated oral administration of 1000 mg/kg of soybean extract on alcohol.

The data of table 3 allow us to conclude that soy extract significantly reduces the consumption of alcohol. Reducing alcohol consumption remains stable during all 7 days of testing and then calms down after impact. Moreover, it can be seen that the reduction in alcohol consumption is more significant compared to the effect which is the sum of effects of individual components of the extract (synergies).

Further, the present invention also relates to a pharmaceutical composition, which contains but with the pharmaceutical composition, containing the extract and intended for the prevention or treatment of premenopausal and postmenopausal symptoms, the pharmaceutical composition containing the extract and intended for the prevention and treatment of breast cancer in women and prostate in men, and pharmaceutical composition containing the extract and intended for the prevention or treatment of alcoholism.

Products or extracts according to the invention can be prepared in a suitable manner in the form of tablets, soft or hard gelatin capsules, granular powder suitable for the preparation of ready-to-use solutions, or liquid forms in accordance with the solubility of these products. Doses of the extract according to the invention are in the range of 30 to 500 mg, in single or multiple daily administration, preferably 200 mg with the introduction of twice a day. A suitable method of administration is oral way.

The following examples illustrate the present invention.

Example 1. Getting soy extract, characterized by a 15% weight content of isoflavones and the ratio 1:1.5 glucosidic isoflavones flour, containing 0.2% glucosidic of isoflavones and 0.3% saponins soy group were subjected to boiling under reflux five times with 30 l of 95% ethanol. The resulting alcoholic extracts were mixed and concentrated to a volume of 5 liters under reduced pressure. The concentrate was dissolved in 1.5 liters of water and was extracted four times in 5 l of n-hexane. The hexane phase was removed and concentrated alcohol phase was extracted four times using 2.5 l of n-butanol. The organic phase was concentrated and dried under reduced pressure. Received 133 g of extract containing 15% isoflavones by weight and 22.5% saponins soy groups In weight.

HPLC chart of the extract obtained using the steps described in this example, the procedure shown in the drawing.

Example 2. Getting soy extract, characterized by a 15% weight content of isoflavones and the ratio 1:1.5 glucosidic of isoflavones and saponins soy group, by treatment with the use of polistirolovoy resin.

An aqueous extract obtained according to example 1 was not subjected to extraction in n-butanol and treated polyethoxylated castor oil (Cremophorand dissolved in the resinous residues by kontsentrirovannyi XAD1180. Then the column is sprinkled with water to remove salts, carbohydrates and surface-active agents, and then elute approximately 10 l of 95% ethanol. After concentration and drying alcohol eluate obtain 130 g of extract, characterized by the same structure as that of the extract obtained according to example 1.

Example 3. Getting soy extract, characterized by a 43% weight content of isoflavones and a 1:1 ratio glucosidic of isoflavones and saponins soy group Century.

20 g of the extract obtained in examples 1 or 2, are suspended in 1 l of aqueous 20% solution of ethyl alcohol and diluted in 0.5 l of ethyl acetate. This suspension is subjected to heated in counterflow with a powerful stirring until then, until complete dissolution, and then leave overnight. Besieged in the saponins (38 g: purity 93%) was isolated by filtration and then share ethylacetamide and aqueous mother liquor. The organic phase is concentrated under reduced pressure and dried. Isoflavonones the remainder (37 g: purity - 81%) dissolved in 1 l of ethanol and mixed with 32 g crystallizing saponins with the purpose of obtaining a product that would contain the situation and poluchaut 69 g of extract, characterized by a 43% weight content glucosidic of isoflavones and 43% by weight of saponins soy group Century.

Example 4. Getting hard gelatin capsules containing soy extract, mg:

Soy extract obtained according to example 1 - 200,0

Lactose is 67.5

Microcrystalline cellulose - 22,5

Colloidal silicon dioxide - 3,0

"Crosscarmellose" sodium (cross-cross-linked polymer of carboxymethylcellulose sodium) - 21,0

Talc - 8,0

Magnesium stearate - 3,0

Example 5. Preparation of tablets containing soy extract, mg:

Soy extract obtained according to example 3 - 400,0

Polysaccharides - 155,5

Microcrystalline cellulose - 57,0

The hypromellose to 12.0

Gidrirovannoe vegetable oil is 19.5

Colloidal silicon dioxide - 3,0

Magnesium stearate - 3,0 e

1. Soy extract, characterized in that it contains from 0.6 to 1.5 weight. including saponins soy group B per 1 weight. including glucosidic of isoflavones, the content glucosidic of isoflavones in the extract is at least 13%.

2. Soy extract on p. 1, characterized in that it contains 1 weight. including saponins soy group B in the calculation is that it includes the following stages: a) extraction of Mature whole soya beans or oil-free soya flour, contains saponins soy group and glucoside isoflavones in reciprocal ratio of 3: 2-2: 3, with the use of aliphatic alcohols or mixtures of these alcohols with water; b) concentrating the extract obtained in stage (a); (C) purification of the concentrated extract obtained in stage (b), from oily and lipophilic components by processing aliphatic hydrocarbons; (d) the extraction of active components using is not miscible with water, aliphatic alcohols; (e) concentrating the extract obtained in stage (d), and its drying.

4. The method according to p. 3, characterized in that after stage (b) or after stage (C) concentrated alcohol extract is subjected to the next stage (d'), followed by stage (e): d') adsorption of active ingredients on polistirolovoy adsorption resin, washing the resin with water and elution of active ingredients ethyl alcohol.

5. The method according to p. 3 or 4, characterized in that after stage (f) carry out the following additional steps: (f) the suspension of the extract obtained in stage (e), in a mixture miscible with water, alcohol and water and breeding it with the use of immiscible aprotic solvent; (g) heating smgr; h) selection besieged saponins soy group filtering; (i) separating the organic phase from the aqueous phase and the concentration of the organic phase with subsequent drying her with getting isoflavonoid components and j) mixture of saponins obtained in stage (h), and isoflavones obtained in stage (i), to obtain a final extract.

6. Pharmaceutical composition containing as the active component of the extract on p. 1 or 2.

7. The pharmaceutical composition according to p. 6, intended for the prevention or treatment of premenopausal and postmenopausal symptoms.

8. The pharmaceutical composition according to p. 6, intended for the prevention or treatment of cancer.

9. The pharmaceutical composition according to p. 6, intended for the prevention or treatment of breast cancer in women.

10. The pharmaceutical composition according to p. 6, intended for the prevention or treatment of prostate cancer in men.

11. The pharmaceutical composition according to p. 6, intended for the prevention or treatment of alcoholism.

 

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