JP7745288B2 - Composition for preventing and/or improving menopausal symptoms - Google Patents

Description

The present invention relates to a composition for preventing and/or ameliorating menopausal symptoms.

Menopause generally refers to the roughly 10 years before and after menopause, which is said to begin around the age of 50 for Japanese women. During menopause, women may experience symptoms specific to this time (menopausal symptoms), such as a sudden flush or heat in the face. These various unpleasant symptoms of menopause are known to be closely related to estrogen, a female hormone, and are thought to occur when the body is unable to cope with the sudden decrease in estrogen levels.

Isoflavones are believed to be effective in improving these menopausal symptoms, and for example, it has been proposed to improve pre- and post-menopausal symptoms (menopausal symptoms) using a composition containing isoflavones extracted from soy isoflavones, etc. (see Patent Document 1). It has also been proposed to improve female hormone disorders, including menopausal disorders, by combining soy isoflavones with theanine and royal jelly (see Patent Document 2).

However, the effect is not sufficient, and there is a need for foods or pharmaceuticals that have sufficient preventive and ameliorative effects against menopausal symptoms and are highly safe.

Japanese Patent Application Publication No. 11-221048 Japanese Patent Application Laid-Open No. 2003-002831

The present invention was made in light of these circumstances, and its objective is to provide an oral composition that is free from side effects, can be taken over a long period of time, and has excellent effects in preventing and/or alleviating menopausal symptoms.

After extensive research into foods and pharmaceuticals that have no side effects and more effectively prevent and/or alleviate menopausal symptoms, the inventors discovered that a combination of specific plant materials enhances the effects of isoflavones in alleviating menopausal symptoms, leading to the completion of the present invention.

That is, the present invention is as follows.
[1] An oral composition comprising isoflavones and plant materials.
[2] The oral composition according to [1], [6] or [7], characterized in that the plant material contains at least one selected from young barley leaves, kale, mugwort, longevity grass, bitter melon, bracken fern, celery, angelica tree, mulberry and spinach.
[3] The oral composition according to [1] or [2], which is used for preventing and/or improving menopausal symptoms.
[4] An oral composition for preventing and/or improving menopausal symptoms, which contains isoflavones as active ingredients, and further contains young barley leaves, kale, and at least one plant material selected from mugwort, longevity grass, bitter melon, Japanese bamboo grass, celery, angelica tree, mulberry, and spinach.
[5] The oral composition according to any one of [1] to [4], which is a functional food.
[6] An oral composition for activating estrogen receptors, comprising isoflavones and plant materials.
[7] An oral composition for relieving sudden hot flashes and redness of the face due to menopause, characterized by containing isoflavones and plant materials.

The oral composition of the present invention has excellent effects in preventing and/or alleviating menopausal symptoms.

FIG. 1 shows the change in estrogen-like activity when the composition of the present invention (isoflavones + young barley leaves + kale + other plant materials) is applied to human breast adenocarcinoma-derived cells.

The oral composition of the present invention is characterized by containing isoflavones and plant materials, and particularly preferably contains isoflavones and at least one plant material selected from mugwort, choumeisou, bitter melon, kumazasa, celery, angelica tree, mulberry, and spinach (mugwort through spinach are sometimes referred to as "other plant materials"). The other plant materials used together with isoflavones, barley leaves, and kale may be used alone (for example, mugwort alone) or in combination of two or more. When using a combination of two or more other plant materials, it is preferable to combine plant materials that are highly effective in enhancing the effects of isoflavones.

Because the specific combination of barley grass, kale, and other plant materials enhances the estrogenic activity of isoflavones, the oral composition of the present invention can effectively prevent and/or improve symptoms characteristic of menopausal women caused by a decline in estrogen.

[Isoflavone]
The isoflavones used in the oral composition of the present invention are flavonoid compounds found in large amounts in legumes, particularly soybeans. The isoflavones of the present invention may be either aglycones or glycosides, with glycosides being preferred. Their origin is not particularly limited, but soy isoflavones derived from soybeans are preferred. Isoflavones are found, for example, in typical soybean processed foods, with average aglycone-equivalent contents per 100 g of soybeans being 140.4 mg, soybean flour 266.2 mg, natto 73.4 mg, and soy milk 24.8 mg. While the oral composition of the present invention does not preclude the use of such isoflavone-containing plants or their processed products, it is preferred to use isoflavones extracted, concentrated, or purified from isoflavone-containing plants or their processed products, or synthetic isoflavones. For example, foods for specified health uses and foods with functional claims must display the recommended intake amount of the active ingredient in the product and provide appropriate dosage and administration instructions. The use of extracts, etc., makes it easier to adjust the effective amount in the product.

[Young barley leaves]
Barley leaves are young leaves of barley, a grass family plant, and may include stems. The form of the barley leaves is not particularly limited, and may be, for example, the leaves as they are harvested, or the leaves may be further processed after harvesting. Examples of such processing include dried powder processing, shredded processing and subsequent dried powder processing, juice extraction and subsequent dried powder processing, and extract extraction and subsequent dried powder processing, and processing methods employed in the art can be applied to these. That is, processed products include pulverized barley leaves, juice, extracts, etc., and pulverized products include powders, granules, etc. Examples of methods for obtaining a pulverized product include a method in which barley leaves and/or stems are dried, coarsely pulverized, heated at 110°C or higher, and then finely pulverized (see JP 2003-033151 A), and a method in which barley leaves and/or stems are blanched, dried, and then pulverized (see JP 2002-065204 A). The squeezed juice or extract may be in liquid form, but can also be used as a paste or dry powder. The extract can be obtained by extraction using an appropriate solvent, and the solvent can be, for example, a water-soluble solvent such as water, ethanol, or aqueous ethanol.

[Kale]
Kale is a plant of the Brassicaceae family, and the part of the plant used as the plant material of the present invention is preferably the green leaves, and may also include the stems. The variety of kale is not particularly limited, and various types of kale can be used, such as kitchen kale, tree kale, bush kale, marrow kale, collard greens, and green-leaf kale. The form of kale is not particularly limited, and for example, the kale may be in the state in which the stems and leaves are harvested, or may be further processed after harvesting. Examples of processed products include pulverized products, squeezed juice, extracts, etc., and examples of pulverized products include powder and granules. The squeezed juice and extracts may be in liquid form, but can also be used as a paste or dried powder. Extracts can be obtained by extraction using an appropriate solvent, and examples of such solvents include water, ethanol, and aqueous ethanol. Kale that has been processed, such as dried and extracted, immediately after harvest is particularly preferred. If time is required before processing, it is preferable to store the kale using storage methods commonly used by those skilled in the art, such as low-temperature storage, to prevent deterioration.

[Plant materials other than young barley leaves and kale (other plant materials)]
In the present invention, the other plant material may be the plant material itself, or may be a processed product such as crushed material, juice, or extract. Examples of crushed material include powder and granules. The juice or extract may be in liquid form, but can also be used as a paste or dry powder. The extract can be obtained by extraction using an appropriate solvent, and examples of the solvent that can be used include water-soluble solvents such as water, ethanol, and aqueous ethanol.
Each plant material will be described below.

(Mugwort)
Artemisia is a plant of the Asteraceae family, and the parts to be used as the plant material of the present invention are preferably green leaves, and may also include stems. The variety of mugwort is not particularly limited, and examples that can be used include artemisia princeps, artemisia absinthium, tarragon, nitro artemisia, giant artemisia, artemisia capillaris, artemisia mandarin, sea artemisia, Japanese ginseng, artemisia annua, artemisia umbellata, Japanese artemisia, artemisia serrata, artemisia serrata, artemisia umbellata, artemisia capillaris, artemisia umbellata, artemisia shikotan, artemisia umbellata, artemisia iwayomogi, artemisia dwarf, artemisia cotton, artemisia kasho, artemisia hitotsuba, artemisia chishima, artemisia broadleaved, artemisia broadleaved, artemisia snowbell, artemisia japonica, artemisia giantum, artemisia westernum, artemisia chinensis, artemisia serrata, artemisia umbell ...

(bitter melon)
Bitter melon is a plant of the Cucurbitaceae family, and is widely distributed in tropical regions from Southeast Asia to southern China, Africa, and South America. The part of the plant used as the plant material of the present invention is preferably the fruit, and may also contain seeds and pulp. Bitter melon has long been widely used in China and Southeast Asia as a medicinal vegetable for stomachic and immune disorders, and various effects such as appetite stimulation and anti-inflammatory and analgesic properties have been identified.

(long life grass)
Chomeisou is a plant of the Umbelliferae family, and the part used as the plant material of the present invention is preferably the green leaves, and may include the stems. Chomeisou is not particularly limited as long as its scientific name is Peucedanum japonicum or Peucedanum japonicum Thumb. Chomeisou is also known locally as Botanboufuu, Chomeisou, Chomeigusa, Chomifusa, Bofuu, Sakuna, Upbasafuna, Chomigusa, Botanboufu, etc.

(Kumazasa)
Kumazasa (Japanese bamboo grass) is a grass family plant, and the part of the plant used as the plant material of the present invention is preferably the green leaves, and may also include the stems. Its leaves have been used as medicine since ancient times. The Shennong Bencaojing describes them as effective for burns, vomiting blood, bloody stools, and urinary congestion. In Japan, they are used as a natural food preservative and packaging agent, as well as a medicine for gastrointestinal diseases, diabetes, high blood pressure, asthma, and colds.

(celery)
Celery is a plant of the Umbelliferae family and is believed to have originated in Europe. The part of the plant used as the plant material of the present invention is preferably the green leaves, and may also include the stems. In ancient Rome and Greece, celery was considered, like ginseng, to have medicinal properties for strengthening the aphrodisiac system and regulating the intestines, and was also said to be effective in relieving fatigue, improving skin condition, and treating diabetes.

(Ashitaba)
Angelica keiskei (Ashitaba) is a plant of the Apiaceae family, and the part used as the plant material of the present invention is preferably the green leaves, and may also include the stems. As a green and yellow vegetable, Angelica keiskei is rich in minerals and vitamins and is said to have effects such as anti-constipation, diuretic effects, and tonic effects. Furthermore, Angelica keiskei is known to contain multiple compounds belonging to the chalcone and coumarin classes, and these compounds have been reported to have effects such as anti-cancer, anti-ulcer, anti-thrombotic, antibacterial, and anti-AIDS effects.

(mulberry)
Mulberry is a plant of the Moraceae family, and the parts to be used as the plant material of the present invention are preferably green leaves, and may also include stems.

(spinach)
Spinach is a plant of the Chenopodiaceae family, and the parts used as the plant material of the present invention are preferably green leaves, and may also include stems.

[Oral composition of the present invention]
The oral composition of the present invention can be used, for example, as a pharmaceutical product (including quasi-drugs), a so-called health food such as a functional food whose efficacy has been approved by a designated organization, such as a food for specified health uses, a food with nutrient function claims, or a food with functional claims, or as a general food, a food additive, or a feed.

The oral composition of the present invention can be used to prevent and/or improve menopausal symptoms. Such oral compositions for preventing and/or improving menopausal symptoms contain isoflavones, young barley leaves, kale, and other plant materials, and are not particularly limited as long as they are distinguishable from other products in that they are used to prevent and/or improve menopausal symptoms. For example, the scope of the present invention includes products that display on the product itself, packaging, instructions, or promotional materials that they are effective in preventing and/or improving menopausal symptoms.

The composition for preventing and/or ameliorating menopausal symptoms of the present invention is not limited to those labeled on product packaging, etc., with the combined components of the present invention (isoflavones, young barley leaves, kale, and other plant materials) as the active ingredients for preventing and/or ameliorating menopausal symptoms. For example, the active ingredients may not be specified. Alternatively, the composition may label only isoflavones as the active ingredient. That is, one form of the oral composition for preventing and/or ameliorating menopausal symptoms of the present invention is an oral composition for preventing and/or ameliorating menopausal symptoms containing isoflavones as active ingredients, further containing young barley leaves, kale, and at least one plant material selected from mugwort, chomeisou grass, Angelica keiskei, kumazasa, mulberry, and spinach.

Specific examples of oral compositions for preventing and/or ameliorating menopausal symptoms of the present invention include pharmaceuticals (including quasi-drugs) and so-called health foods. Examples of so-called foods include those labeled as having an effect of preventing and/or ameliorating menopausal symptoms, such as "for menopausal worries," "menopausal measures," "for women concerned about menopausal symptoms," "support for women's age-related worries," "restoring the body balance of menopausal women," "relieving menopausal fluctuations in menopausal women," "for facial hot flashes in older women," "for the health of women in the menopausal generation," and "reducing sudden hot flashes and redness in the face."

The oral composition of the present invention may be in the form of, for example, a tablet, capsule, powder, granules, liquid, granules, rod, plate, block, solid, round, paste, cream, caplet, gel, chewable, or stick. Among these, tablet, capsule, powder, granule, and liquid forms are particularly preferred. Specific examples include supplements, packaged beverages filled in PET bottles, cans, jars, etc., instant beverages that are dissolved in water (hot water), milk, fruit juice, green juice, etc., and food additives. These are preferred because they are easy to consume during meals and can enhance palatability.

The oral composition of the present invention may contain isoflavones, young barley leaves, kale, and other plant materials (components of the present invention) in amounts appropriate to the extent that their effects are achieved.

In general, when the oral composition of the present invention is a pharmaceutical or supplement, the components of the present invention are preferably contained in an amount of 0.1 to 100% by mass, more preferably 1 to 85% by mass, and even more preferably 5 to 70% by mass, calculated on a dry mass basis.

In order to more effectively exert the effects of the present invention, the components of the present invention preferably account for 80% or more of the total oral composition of the present invention, calculated on a dry mass basis, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 100%.

The mass ratio of isoflavones to the total of barley leaves, kale, and other plant materials in the oral composition of the present invention is preferably in the range of 1:0.001 to 1:1000, more preferably 1:0.01 to 1:500, and even more preferably 1:0.1 to 1:200, calculated on a dry mass basis. There are no particular limitations on the ratio of barley leaves, kale, and other plant materials, but it is preferable that each component be present in approximately the same amount.

There are no particular restrictions on the amount of intake of the oral composition of the present invention, but from the perspective of more significantly exerting the effects of the present invention, it is preferable for an adult to ingest at least 10 mg/day of isoflavones (equivalent to aglycones), more preferably at least 15 mg/day, and even more preferably at least 20 mg/day. The upper limit is, for example, 75 mg/day, preferably 70 mg/day. The oral composition of the present invention is particularly preferably ingested by women who have reached menopause.

The oral composition of the present invention can be stored as a daily dose in a single container or divided into, for example, two or three containers so that the daily intake amount is the above-mentioned amount.

The oral composition of the present invention can be produced by known methods, if necessary, by adding orally acceptable ingredients other than the ingredients of the present invention. Examples of ingredients other than the ingredients of the present invention include vitamins such as water-soluble vitamins (vitamins B1, B2, B3, B5, B6, B12, B13, B15, B17, biotin, choline, folic acid, inositol, PABA, vitamin C, vitamin P) and oil-soluble vitamins (vitamins A, D, E, K); minerals such as calcium, magnesium, phosphorus, and iron; sulfur-containing compounds found in taurine and garlic; flavanoids or flavonoids such as hesperidin and quercetin; proteins such as collagen; peptides; amino acids; animal fats and oils; vegetable fats and oils; and ground products or extracts of animals and plants.

The present invention will now be described with reference to examples.

[Example 1]
1. Preparation of test substance [isoflavone]
As the isoflavone, an isoflavone (aglycone) mixture (derived from soybean; 093-04771, manufactured by Wako Pure Chemical Industries, Ltd.) was used.

The isoflavone mixture was dissolved in DMSO to a concentration of 20 μg/mL. It was then diluted 200-fold with the medium used to prepare the test substance, and adjusted to a concentration of 0.1 μg/mL (DMSO concentration: 0.5%). Equal volumes of the 0.1 μg/mL isoflavone mixture and medium with a DMSO concentration of 0.5% were added to give a final concentration of 0.05 μg/mL isoflavone mixture and a final DMSO concentration of 0.5%.

[Young barley leaves, kale, and other plant materials]
(Pretreatment)
For plant materials other than young barley leaves, kale, and bitter melon, the green leaves (and stems) and the bitter melon fruits were harvested and washed with water to remove any adhering mud, and the long ones were cut into pieces of about 5 to 10 cm in length for pre-processing.

For young barley leaves, kale, and other plant materials, the cut plant bodies of young barley leaves, kale, mugwort, chomeisou, bitter melon, kumazasa, celery, angelica tree, and spinach were blanched once in a blanching tank using a fast-acting steamer at 90-100°C for 90-120 seconds, and then cooled in cold water. Mulberry was steamed in steam at 98-100°C, and then cooled in cold water.

Tea leaves (green leaves of the tea plant, scientific name: Camellia sinensis) were used for comparison with other plant materials. The picked tea leaves were steamed at 98-100°C and then cooled in cold water.

The cooled young barley leaves, kale, and other plant materials, as well as the tea leaves, were dried using a dryer for 20 to 50 minutes with hot air at 80 to 130°C so that the moisture content was 5% by mass or less. After drying, the green leaves were coarsely crushed to a size of approximately 5 mm, and then finely crushed using a jet mill so that 90% passed through a 200 mesh filter, yielding green leaf powder and fruit powder.

(Preparation of test substance)
The obtained green leaf powder, etc. was suspended in a medium for preparing test substances and suspended in a vortex mixer for approximately 30 minutes. After filter sterilization, the barley leaf powder, kale, and other plant materials suspended in the medium were mixed at a ratio of 1:1:1, and serially diluted so that the total of the three types was 2x concentration. The desired concentrations were achieved by adding half of the total 2x concentration soy isoflavones and half of the 2x concentration mixture of barley leaf, kale, and other plant materials.

2. Cell Culture Human breast adenocarcinoma-derived cells (MCF-7) were cultured in 10% FBS-1% NEAA-DMEM in a 75 ^cm2 flask in a 37°C, 5% _CO2 incubator. The cells were then suspended by trypsinization and seeded from the 75 ^cm2 flask into each well of a 96-well plate at a cell density of 2 x ¹⁰³ cells/well. The cells were pre-cultured for 24 hours in a 37°C, 5% _CO2 incubator.
Furthermore, the detection system for estrogenic activity using the proliferation of MCF-7 cells as a marker is a stable detection system, and since MCF-7 cells have estrogen receptors, the estrogenic activity of a substance can be confirmed by the binding of the substance to the receptor and the promotion of cell proliferation.

After removing the medium from each well, the wells were washed once with EMEM medium (serum-free). 100 μL of medium containing the test substance adjusted to a predetermined concentration was added to each well, and the wells were cultured in a CO ₂ incubator for 2 days.

For the combined test substance and isoflavone samples, the isoflavones were adjusted to a 2x concentration and the test substance-containing medium to a 4x concentration, and the target concentrations were achieved by adding 50 μL of isoflavones and 25 μL of test substance-containing medium. The medium used was 10% FBS (charcoal-treated) - 1% NEAA - 1 mM Glutamine - EMEM.

Controls included a control (+) in which isoflavones were adjusted to a final concentration of 0.05 μg/mL, and a control (-) in which no isoflavones were added.

3. Measurement of cell activation activity After culturing for 2 days, the medium was removed, the cells were washed with serum-free DMEM, and 150 μL of Cell Counting Kit-8 solution (Dojindo Laboratories, Inc.) diluted 30-fold with serum-free DMEM was added. The cells were then placed in a 37°C, 5% _CO2 incubator to allow for adequate color development, and the absorbance at 450 nm was measured. Based on the obtained data, the % of control was calculated using the following formula.

% of control
= (Data sample-Data blank)/(Data control-Data blank)×100

The results are shown in Figure 1. Figure 1 shows the change in estrogenic activity when the value for the isoflavone-only addition group (0.05 μg/mL) is set to 0. Groups other than the control group contained young barley leaves, kale, and other plant materials. For example, "Yomogi 125" contains young barley leaves, kale, and mugwort, each at a concentration of 41.7 μg/mL. A "-" indicates that no isoflavones were added, and a "+" indicates that isoflavones (0.05 μg/mL) were added.

As shown in Figure 1, estrogenic activity was enhanced by combining isoflavones with the specific plant materials of the present invention, which have low or no estrogenic activity on their own. In particular, when barley, kale, and mugwort were combined, estrogenic activity was enhanced more than with other combinations. On the other hand, when barley, kale, and tea were combined, not only did the plant materials themselves not exhibit estrogenic activity, but their estrogenic activity was inhibited even when combined with isoflavones. These findings demonstrate that the composition of the present invention can effectively prevent and/or ameliorate menopausal symptoms.

Example 2 (Production of tablets)
Soybean-derived isoflavones (as aglycones) 25mg
Barley grass powder 25mg
Kale powder 25mg
Mugwort powder 25mg
Reduced maltose 50mg
Calcium stearate 100mg
The mixture was compressed into two tablets.

[Example 3] (Production of capsules)
Soybean-derived isoflavones (as aglycones) 50mg
Barley grass powder 25mg
Kale powder 25mg
Mugwort powder 25mg
Shell calcium 50mg
The mixture was encapsulated in a soft capsule to produce one capsule.

[Example 4] (Production of granules)
Soybean-derived isoflavones (as aglycones) 75mg
Barley grass powder 1000mg
Kale powder 500mg
Spinach powder 135mg
Lactose 50mg
The mixture consisting of the remaining reduced maltose was mixed and then granulated to produce 3 g of granules.

The oral composition of the present invention can be used as a pharmaceutical or health food that is highly effective in preventing and/or ameliorating menopausal symptoms, making the present invention highly industrially useful.

Claims (1)

A food composition for preventing and/or improving menopausal symptoms, characterized by containing isoflavones, crushed barley young leaves , crushed kale powder , and crushed mulberry leaves (excluding the food compositions (1) and (2) described below).
(1) Excluding food compositions containing plant fermentation powder, Coleus forskohlii extract powder, L-carnitine, yeast, and koji
(2) Excluding soy milk and food compositions containing vitamin C