JP7565172B2 - Female hormone-like activity enhancer - Google Patents

Description

The present invention relates to an agent that enhances female hormone-like activity.

Women's health is greatly influenced by two types of female hormones, estrogen (follicular hormone) and progesterone (luteal hormone), which are secreted from the ovaries. A deficiency of female hormones can cause symptoms such as menopause and osteoporosis. Hormone replacement therapy (HRT) is widely used as a treatment to alleviate these symptoms. However, because HRT is a treatment that artificially replenishes large amounts of female hormones, while it is highly effective, it also places a heavy burden on the body and makes it very difficult to adjust the dosage.

In recent years, plant-derived isoflavones have been attracting attention as components that mimic female hormones. It is believed that this female hormone-like effect is due to the similarity of the chemical structure of isoflavones to that of estrogen. In particular, equol, which is derived from isoflavones contained in soybeans, is known to have estrogenic activity (Non-Patent Documents 1 and 2).

Sathyamoorthy, N. and Wang, T. T. , Eur. J. Cancer, 33, 2384-2389 (1997) Schmitt, E. et al. , Toxicol. In Vitro, 15, 433-439 (2001)

Although equol is known to have a female hormone-like effect, no agent capable of enhancing this effect is known. Therefore, the present invention aims to provide an agent capable of enhancing the female hormone-like effect of equol.

After extensive research, the inventors discovered that combining equol with carrot extract in a specific ratio enhances the female hormone-like action. The present invention was completed based on this finding and through further research.

That is, the present invention provides the following aspects.
Item 1. A female hormone-like activity enhancer comprising equol and carrot extract, the content of the carrot extract being 2.4 to 60 parts by weight per part by weight of the equol.
Item 2. The female hormone-like activity enhancer according to Item 1, wherein the carrot extract is a hydroethanolic extract of carrot.
Item 3. The female hormone-like agent according to Item 1 or 2, which enhances estrogen activity.

The present invention provides an agent that can enhance the female hormone-like action of equol, which is expected to improve various symptoms caused by a decrease in female hormones.

The female hormone-like activity enhancer of the present invention is characterized by containing equol and carrot extract in a specific ratio. The female hormone-like activity enhancer of the present invention is described in detail below.

Equol Equol is a substance produced by the metabolism of daidzein, a type of isoflavone, by intestinal bacteria and digestive enzymes in the body, and specifically refers to 7-hydroxy-3-(4'-hydroxyphenyl)-chroman. Equol is known as a component having female hormone-like activity.

Equol may be synthetic equol obtained by chemical reduction of daidzein (daidzein glycoside, daidzein and/or dihydrodaidzein), or it may be a metabolite of equol-producing bacteria from a raw material containing daidzein (daidzein glycoside, daidzein and/or dihydrodaidzein).

Raw materials containing daidzeins are not particularly limited, and examples thereof include plants (specifically, soybeans, kudzu, kudzu root, red globe, alfalfa, etc.), processed products thereof (specifically, soybean flour, boiled soybeans, tofu, fried tofu, soy milk, soybean hypocotyls, soybean hypocotyl extracts, etc.), and fermented preparations thereof (specifically, natto, soy sauce, miso, tempeh, fermented soy beverages, etc.). In addition, the equol-producing bacteria may be any microorganism that can assimilate daidzeins to produce equol, and examples thereof include microorganisms belonging to the genus Lactococcus, the genus Streptococcus, and the genus Bacteroides.

In the present invention, equol may be blended using a fermentation culture of an equol-producing bacterium that uses raw materials containing daidzeins, or equol, which is a metabolite, may be isolated and purified from the fermentation culture.

The content of equol in the female hormone-like activity enhancer of the present invention may be set depending on the content of other ingredients to be blended, the form of the female hormone-like activity enhancer, the amount to be taken per serving, etc., and may be, for example, 0.01 to 70% by weight, preferably 0.05 to 60% by weight, more preferably 0.1 to 55% by weight, and even more preferably 0.1 to 50% by weight. In the present invention, when a fermentation culture is used to blend equol, the fermentation culture is used so that the equol content is the above-mentioned equol content.

Ginseng extract Ginseng (ginseng) is the root of Panax ginseng C.A. Meyer (Panax schinseng Nees) of the Araliaceae family, with the fine roots removed, or the roots lightly boiled, and is used as a herbal medicine (Japanese Pharmacopoeia) mainly for the purposes of tonics, digestive stimulants, antidiarrheal drugs, tranquilizers, hypoglycemic drugs, cardiac stimulants, heat retaining drugs, anti-fatigue drugs, etc. Incidentally, ginseng is both the name of the herbal medicine (Japanese Pharmacopoeia) and the name of the plant. Ginseng is commercially available from Nippon Funa Yakuhin Co., Ltd., Tochimoto Tenkaido Co., Ltd., etc.

Carrot extract is described in the quasi-drug raw material standard and is publicly known. Specifically, carrot extract can be obtained by extracting fine roots of carrot or a fermented product thereof with an extracting solvent. In the present invention, it is preferable to use an extract obtained by extracting fine roots of carrot with an extracting solvent as the carrot extract. Examples of the extracting solvent used in the carrot extracting process include water and/or ethanol. From the viewpoint of further enhancing the effect of enhancing female hormone-like activity, preferred are ethanol or aqueous ethanol, more preferred are aqueous ethanol, and even more preferred are aqueous solutions containing 55 to 80 v/v% (preferably 57 to 70 v/v%, more preferably 59 to 65 v/v%) ethanol. Carrot extract is commercially available from, for example, Nippon Funa Yakuhin Co., Ltd., Matsuura Yakugyo Co., Ltd., Alps Yakuhin Kogyo Co., Ltd., etc.

The carrot extract may be a commercially available product, or may be an extract liquid obtained by extracting the above-mentioned carrot parts using the above-mentioned extraction solvent and concentrating the obtained extract liquid as necessary, or may be a dried extract powder obtained by drying the extract liquid. The method for drying the carrot extract is not particularly limited, and includes known methods such as a spray drying method and a method in which a suitable adsorbent (e.g., silicic anhydride, starch, etc.) is added to a soft extract with a high concentration of the extract liquid to obtain an adsorbed powder.

In the female hormone-like activity enhancer of the present invention, the ratio of the equol content to the carrot extract content is 2.4 to 60 parts by weight, calculated as the content of the carrot extract per 1 part by weight of equol. From the viewpoint of further enhancing the effect of enhancing the female hormone-like activity by combining with carrot extract, the content of the carrot extract per 1 part by weight of equol is preferably 4 to 48 parts by weight, more preferably 10 to 40 parts by weight, even more preferably 16 to 32 parts by weight, and even more preferably 22 to 28 parts by weight.

The content of carrot extract in the female hormone-like activity enhancer of the present invention may be set so that the ratio to the content of equol falls within the above range, for example, 0.3 to 10% by weight, preferably 0.8 to 5% by weight.

In the present invention, when a liquid extract such as an extract liquid or soft extract is used in the formulation of carrot extract, the amount remaining after excluding the solvent contained in the liquid extract is used so that it corresponds to the ratio and content of the above-mentioned carrot extract, and when a dry extract powder containing an additive such as an adsorbent is used in the formulation of carrot extract, the amount remaining after excluding the additive contained in the dry extract powder is used so that it corresponds to the ratio and content of the above-mentioned carrot extract.

Other components : The female hormone-like activity enhancer of the present invention may contain other pharmacological components, if necessary, in addition to the above-mentioned components. The types of such pharmacological components are not particularly limited, but examples thereof include antacids, stomachics, digestives, intestinal regulators, antispasmodics, mucosal repair agents, anti-inflammatory agents, astringents, antiemetics, antitussives, expectorants, anti-inflammatory enzymes, sedatives, hypnotics, antihistamines, caffeines, cardiac diuretics, antibacterial agents, vasoconstrictors, vasodilators, local anesthetics, herbal powders, herbal extracts, vitamins, and menthols. These pharmacological components may be used alone or in combination of two or more. The content of these pharmacological components may be appropriately set from known ones depending on the type of pharmacological component to be used.

The female hormone-like activity enhancer of the present invention may contain pharma- ceutically acceptable bases and additives, etc., as necessary, in order to prepare the desired dosage form. Examples of such bases and additives include excipients, binders, disintegrants, lubricants, isotonicity agents, plasticizers, dispersants, emulsifiers, solubilizers, wetting agents, stabilizers, suspending agents, adhesives, coating agents, glossing agents, water, oils and fats, waxes, hydrocarbons, fatty acids, higher alcohols, esters, water-soluble polymers, surfactants, metal soaps, lower alcohols, polyhydric alcohols, pH adjusters, buffers, antioxidants, UV inhibitors, preservatives, flavorings, fragrances, powders, thickeners, dyes, chelating agents, etc. These bases and additives may be used alone or in combination of two or more. The content of these bases and additives may be appropriately set from known ones depending on the type of added component used and the dosage form, etc.

Dosage form The dosage form of the female hormone-like activity enhancer of the present invention is not particularly limited, and may be any of tablets, granules, powders, pills, capsules, films, liquids (drinks), etc. Among these dosage forms, from the viewpoint of ease of administration, etc., preferred are tablets, granules, and films. These dosage forms can be prepared using pharma- ceutically acceptable base materials and additives, and the types and amounts of the base materials and additives are also known in the field of formulation technology.

Uses The female hormone-like activity enhancer of the present invention is used for the purpose of enhancing female hormone-like activity, more specifically, for the purpose of enhancing physiological and pathophysiological actions expressed via female hormone receptors.

The female hormone-like activity enhancer of the present invention enhances the binding activity to the estrogen receptor, and is therefore preferably used for the purpose of enhancing the physiological and pathophysiological actions expressed via the estrogen receptor.

More specifically, the female hormone-like activity enhancer of the present invention is used to prevent or improve symptoms caused by a decrease in female hormones, such as menopausal symptoms, menopausal disorders, osteoporosis, physical condition fluctuations, menstrual pain, and a decrease in skin or hair quality (ability to retain collagen and/or elastin).

Therefore, the female hormone-like activity enhancer of the present invention is applicable to women with reduced female hormones, particularly women who are aware of symptoms caused by reduced female hormones. Specific target subjects include women aged 40 years or older, menopausal women, women with premature ovarian failure after oophorectomy or chemotherapy, women after hysterectomy, elderly women, etc.

Dosage and Usage The female hormone-like activity enhancer of the present invention is administered orally. The dosage of the female hormone-like activity enhancer of the present invention is appropriately set depending on the degree of female hormone reduction, age, etc., and may be, for example, 0.1 to 100 mg, preferably 0.5 to 40 mg, more preferably 1.0 to 10 mg, even more preferably 1.5 to 5 mg, and particularly preferably 1.8 to 3 mg, in terms of the daily intake of equol.

The timing of taking the female hormone-like activity enhancer of the present invention is not particularly limited, and it may be taken before, after, or between meals, but is preferably taken before or between meals.

The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

Test Example As equol, (S)-equol (Wako Pure Chemical Industries, Ltd.) was prepared. As carrot extract, extract powder obtained by extracting ginseng rootlets with 60 v/v % aqueous ethanol solution and drying was used. Test sample solutions (1 v/v % dimethyl sulfoxide) were prepared by dissolving equol and carrot extract in dimethyl sulfoxide at the ratios shown in Table 1 and diluting with culture medium. The culture medium used was prepared by adding 3 v/v % charcoal-treated fetal bovine serum, 100 U/mL penicillin, and 100 pg/mL streptomycin to sodium pyruvate-containing, L-glutamine-free, and phenol red-free Dulbecco's modified Eagle's medium (1.0 g/L glucose) (liquid) so that the L-glutamine concentration was 0.5 mM.

Human breast cancer-derived MCF7 cells (KAC Corporation) were prepared. MCF7 cells are cancer cells whose proliferation ability increases in an estrogen-dependent manner, and the proliferation ability of MCF7 cells is used as a detection system for estrogenic activity.

MCF7 cells suspended in medium were seeded onto a 96-well culture plate at a cell density of 2 x ¹⁰³ cells/well/100 μL. The cells were cultured for one day at 37°C in ₅ v/v% CO2, after which the medium was removed and fresh medium (50 μL/well) was added, and the test sample solution was added (10 μL/well) so that the final concentrations of equol and carrot extract were as shown in Table 1, and the cells were similarly cultured for three days.

After culturing, the absorbance at 450 nm (corresponding to the number of cells) was measured using a cell proliferation/cytotoxicity assay kit (product name "Cell Counting kit-8", Dojindo Research Institute, Inc.). The measured absorbance value at 450 nm was used to calculate the cell proliferation promotion rate according to the following calculation formula. The test was performed with n=3, and the average of the obtained cell proliferation promotion rates was used as the evaluation value of the female hormone-like effect. The results are shown in Table 1.

As shown in Comparative Example 1, the female hormone-like effect of equol alone was insufficient, and as shown in Comparative Examples 2 to 4, carrot extract alone showed a female hormone-like effect when used in an amount more than five times that of equol, but the degree of the effect was insufficient. On the other hand, as shown in Examples 1 and 2, a synergistic enhancement effect of the female hormone-like effect (an effect exceeding the additive effect) was observed by combining equol and carrot extract at a specified ratio. In particular, it was recognized that the synergistic enhancement effect of the female hormone-like effect was further enhanced (the degree of exceeding the additive effect was further increased) by combining equol and carrot extract at the ratio shown in Example 2. On the other hand, as shown in Comparative Example 5, when equol and carrot extract were combined at a ratio exceeding the specified ratio, the result was less than the additive effect of the female hormone-like effect. In other words, it was recognized that the enhancement effect of the female hormone-like effect observed in Examples 1 and 2 is an effect specific to the combination of equol and carrot extract at a specified ratio.

Claims (3)

A female hormone-like activity enhancer comprising equol and a carrot (excluding fermented ginseng) extract, the content of said carrot extract being 22 to 28 parts by weight per part by weight of said equol. The female hormone-like activity enhancer according to claim 1, wherein the carrot extract is a hydroethanolic extract of carrot. The female hormone-like agent according to claim 1 or 2, which enhances estrogen activity.