TWI879838B - Uses of the fermented extract of fresh leaves of evergreen daylily - Google Patents

Abstract

The present invention provides an oral preparation that can effectively improve sleep quality. The present invention prepares an oral preparation for improving sleep quality, which contains a fermented extract of fresh leaves of evergreen daylily (Hemerocallis fulva var. sempervirens) as an active ingredient.

Description

Uses of the fermented extract of fresh leaves of evergreen daylily

The present invention relates to a composition for improving sleep quality.

About one-third of our lives are spent on sleep, so improving sleep quality is important. Data show that sleep is closely related to life rhythm and stress. For example, the "2000 Insurance Benefit Trend Survey" published by the Ministry of Health, Labor and Welfare of Japan pointed out that the greater the stress, the more likely it is to have problems such as "not feeling deep asleep even when waking up in the morning" and "difficulty falling asleep".

Research has been conducted on improving sleep quality by using plant-derived ingredients. For example, Patent Document 1 discloses a sleep quality improver containing an extract of an ash plant or Siberian larch.

Evergreen daylily (Hemerocallis fulva var. sempervirens) (Okinawan name: forget-me-not (pronounced: kwanso), sleep-inducing (pronounced: nibui) grass) belonging to the genus Hemerocallis of the Liliaceae family has been considered to have a sleep-inducing effect since the Ryukyu Dynasty, and has been cooked into "Okinawa tonic soup (pronounced: shinjimun)" with forget-me-not and pork, and eaten as a medicinal dish. Today, evergreen daylily is used in beverages such as tea, snacks, etc., and is also used for viewing because of its beautiful flowers, and has attracted much attention as a representative ingredient of Okinawa. [Prior art literature] [Patent literature]

[Patent Document 1] International Publication No. 2013/051727

[The problem that the invention wants to solve]

It is known that daylily (forget-me-not) contains compounds that are effective in improving sleep. However, in the case of preparing daylily extracts (extracts), the development of technology for efficiently containing compounds that are effective in improving sleep is not sufficient.

Furthermore, when preparing the evergreen daylily extract, no detailed tests have been conducted on what kind of sleep improvement effects can be expected for humans. Although it is said to improve sleep, there are many types of sleep quality, such as shortening the time to fall asleep, increasing the total time of non-rapid eye movement sleep associated with deep sleep, the depth of non-rapid eye movement sleep, reducing mid-morning awakening, being able to get up easily, reducing sleepiness when getting up, and improving fatigue after getting up, and the needs of people who have trouble with sleep quality are different. In other words, when preparing the evergreen daylily extract, clinical trials have not been conducted on what kind of sleep quality can be improved for humans. [Technical means to solve the problem]

In view of the above-mentioned topic, in-depth research was conducted and it was found that by using fresh leaves of evergreen daylilies instead of drying them to prepare fermented extracts, an oral preparation having a significant effect on improving sleep quality could be obtained. Clinical trials on the fermented extracts of fresh leaves of evergreen daylilies were conducted on humans, and it was found that the extracts had a significant effect on sleep quality, especially on reducing the time it takes to fall asleep and increasing deep sleep, thereby completing the present invention.

That is, the present invention provides a composition as shown below. Item 1. An oral preparation for improving sleep quality, which contains a fermented extract of fresh leaves of evergreen daylily as an active ingredient.

Item 2. An oral preparation as described in Item 1, wherein the sleep quality mentioned above refers to a reduction in sleepiness upon waking up, recovery from fatigue upon waking up, a reduction in the time it takes to fall asleep, an increase in non-rapid eye movement sleep, or a reduction in mid-sleep awakenings.

Item 3. The oral preparation as described in Item 1, wherein the water content of the fresh leaves of the evergreen daylily is 10% or more.

Item 4. The oral preparation as described in Item 1, wherein the fresh leaves of the evergreen daylily are obtained from the evergreen daylily within four months after the flowers have fallen.

Item 5. The oral preparation as described in Item 1, wherein the daily intake of the fermented extract of fresh leaves of the evergreen daylily is 100 mg/day to 4000 mg/day in terms of dry solid content. [Effect of the invention]

The use of an oral preparation containing a fermented extract of fresh leaves of evergreen daylily as an active ingredient can improve sleep quality.

[Fresh leaf fermented extract of evergreen daylily] Evergreen daylily (Hemerocallis fulva var. sempervirens) is a plant of the genus Hemerocallis in the family Liliaceae. "Evergreen daylily" is the Japanese name, and in Okinawa Prefecture it is also called forget-me-not grass (pronounced: kuwanso), forget-me-not grass (pronounced: kwanso), and sleep-inducing grass (pronounced: nibuigusa).

There is no specific restriction on the origin of the evergreen daylily. For example, it can be produced in Japan or outside Japan. For example, the Japanese origin includes Okinawa Prefecture, southern Kyushu, and the Minamisai Islands. For example, the non-Japanese origin includes Asian origins such as China and Taiwan.

In the oral preparation of the present invention, from the viewpoint of significantly exerting the effect of improving sleep quality, although the part for extracting the evergreen daylily extract may be the whole plant, or at least one selected from the group consisting of leaves, stems, flower buds, roots and flowers, it is preferred to use at least the leaves of the evergreen daylily.

The inventors of the present invention have newly discovered that if the leaves of the evergreen daylily are dried, the content of oxo-jasmine will be significantly reduced. Therefore, the leaves of the evergreen daylily are preferably fresh leaves (undried).

In one embodiment, in addition to the leaves of the evergreen daylily, the stems and/or flower buds of the evergreen daylily can also be used in combination as the extraction parts. Since the content of oxo-jasmine amino acid is significantly reduced due to the drying step when using these extraction parts, it is better to use these extraction parts in a fresh state.

The inventors of the present invention have found that, in one embodiment, if the flowers of the evergreen daylily bloom from the bud state, the content of oxo-jasmine will be significantly reduced, so it is better to exclude the flowers of the evergreen daylily from the extraction part.

From the perspective of significantly improving sleep quality, the evergreen daylily is preferably obtained by fermenting fresh leaves.

The moisture content of the fresh leaves (undried) of the evergreen daylily can be, for example, 10% or more, preferably 15% or more, more preferably 20% or more, and even more preferably 25% or more. Regarding the above moisture content, when the fresh leaves of the evergreen daylily are commercially available, it is preferably the above moisture content at the time of sale, and more preferably the above moisture content at the time of fermentation treatment.

The inventors of the present invention have newly discovered that the content of oxo-ergoline in fresh leaves of evergreen daylilies may decrease significantly depending on the harvest period. Specifically, it was found that after the flowering period, the content of oxo-ergoline in the leaves increases significantly three to four months after the flowers fall, and if it is more than four months after the flowers fall, the content of oxo-ergoline in the leaves decreases sharply. Therefore, the fresh leaves of evergreen daylilies are preferably harvested within at least four months after the flowers fall, more preferably harvested one to four months after the flowers fall, further preferably harvested two to four months after the flowers fall, and particularly preferably harvested three to four months after the flowers fall.

In the fermentation treatment for obtaining the fermentation product, a fermentation means known to the industry can be used. As the fermentation means, for example, koji, lactic acid bacteria and yeast can be listed. As the fermentation means, yeast, such as yeast, can be preferably used. By performing the fermentation treatment, the decomposition and reduction of oxo-jasmine acid contained in the fresh leaves of the evergreen daylily due to the presence of sugars can be suppressed.

As koji fungi, for example, filamentous fungi belonging to the genus Aspergillus can be cited.

Examples of lactic acid bacteria include bacteria belonging to the genus Lactobacillus, Lactococcus, or Streptococcus.

Examples of yeast include yeast belonging to the genus Saccharomyces, Schizosaccharomyces, Kluyveromyces, or Pichia. From the viewpoint of significantly exerting the effects of the present invention, the yeast is preferably brewing yeast (Saccharomyces cerevisiae) or a subspecies thereof, which is also called yeast.

The conditions of the fermentation treatment can be appropriately selected by the industry. The conditions of the fermentation treatment include, for example, a step of mixing brewing yeast (Saccharomyces cerevisiae) as yeast with at least a portion of fresh leaves of evergreen daylily, preferably with juice squeezed from fresh leaves of evergreen daylily.

The temperature condition for the fermentation treatment may be, for example, 20 to 50°C, preferably 25 to 45°C, more preferably 30 to 40°C, and even more preferably 35 to 38°C.

The fermentation treatment time may be, for example, 20 to 70 hours, preferably 25 to 65 hours, and more preferably 30 to 60 hours.

The fermentation process may also include a sterilization step as appropriate.

The fermentation treatment may also include, for example, a step of soaking at least a portion of the fresh leaves of the evergreen daylily in water. The fresh leaves of the evergreen daylily soaked in water may be subjected to the above-mentioned fermentation means, such as adding yeast, and then subjected to the fermentation treatment.

The immersion time in the immersion step can be set, for example, to 10 minutes to 10 hours, preferably 30 minutes to 8 hours, more preferably 40 minutes to 5 hours, and further preferably 1 hour to 3 hours.

The soaking step may also include a step of boiling the water.

Fresh leaves of evergreen daylily suitable for fermentation treatment may be pulverized before fermentation treatment.

The fresh leaves of the evergreen daylily suitable for fermentation can also be squeezed before fermentation. Preferably, the juice of the fresh leaves of the evergreen daylily that have been squeezed is used for fermentation, but the present invention is not limited thereto.

The fresh leaves of the evergreen daylily suitable for fermentation can also be ground before fermentation. By using the paste of the ground fresh leaves of the evergreen daylily for fermentation, the fermentation efficiency can be improved, for example, it is suitable for preparing a solid preparation, but it is not limited thereto.

The fresh leaves of the evergreen daylily are preferably subjected to a heat treatment to inactivate various enzymes contained in the fresh leaves of the evergreen daylily before or after the fermentation treatment, preferably after the fermentation treatment, but the present invention is not limited thereto. The temperature conditions for the heat treatment may be, for example, 60 to 100° C., preferably 70 to 95° C., and more preferably 75 to 90° C. The time for the heat treatment may be, for example, 10 seconds to 10 minutes, preferably 10 seconds to 5 minutes, and more preferably 10 seconds to 3 minutes.

The fresh leaves of the evergreen daylily are preferably subjected to a freezing treatment to inactivate various enzymes contained in the fresh leaves of the evergreen daylily before or after the fermentation treatment, preferably after the fermentation treatment, in addition to or instead of the above-mentioned heating treatment, but the present invention is not limited thereto. The temperature conditions for the freezing treatment are preferably -4°C or less, more preferably -10°C or less, further preferably -15°C or less, particularly preferably -18°C or less, and most preferably -20°C or less.

By freezing, the fermented extract of fresh leaves of evergreen daylily can be used in frozen foods or frozen desserts, etc. By inactivating various enzymes contained in the fresh leaves of evergreen daylily, foods in which the decomposition of oxo-erucic acid is inhibited can be produced, but the invention is not limited thereto.

As the oral preparation of the present invention, a fermentation extract obtained by subjecting fresh leaves of evergreen daylily to the above-mentioned fermentation treatment can be used, and a commercially available fermentation product can also be used.

In the oral preparation of the present invention, the daily intake of the fermented extract of fresh leaves of evergreen daylily can be set to, for example, 100 mg/day to 4000 mg/day, preferably 200 mg/day to 3500 mg/day, calculated as dry solid content.

In addition, the oral preparation of the present invention can be set to, for example, 2 mg/day to 20,000 mg/day, preferably 2 mg/day to 10,000 mg/day, more preferably 2 mg/day to 2,500 mg/day, and particularly preferably 2 mg/day to 2,000 mg/day, based on the content of oxo-jasmine.

According to the present invention, it is found that the extraction efficiency of components effective for sleep quality can be improved by using the fermentation extract obtained by subjecting the fresh leaves of evergreen daylily to the above-mentioned fermentation treatment. However, in one embodiment, it is preferred that the content of oxo-jasmine in 100 g of the fermentation extract of the fresh leaves of evergreen daylily is 2 mg or more, for example, it can be set to 2 mg to 20000 mg, 2 mg to 10000 mg, 2 mg to 2500 mg, 2 mg to 2000 mg, etc.

[Use] The oral preparation of the present invention is used to improve sleep quality. In this manual, "improvement" refers to the improvement of sleep quality, the prevention or delay of deterioration of sleep quality.

Sleep quality can refer to the overall sleep quality during sleep or the sleep quality of a part of the sleep period. The best sleep is the first sleep cycle, which allows the brain to rest efficiently. Sleep has rapid eye movement sleep and non-rapid eye movement sleep, and rapid eye movement sleep and non-rapid eye movement sleep alternate during sleep. Sleep cycle refers to the period from the end of a rapid eye movement sleep to the end of the next rapid eye movement sleep. The first sleep cycle refers to the first sleep cycle after falling asleep.

The above sleep quality refers to the observation of one or more of the following phenomena: reduction of sleepiness upon waking up, recovery from fatigue upon waking up, reduction of time to fall asleep, increase of non-rapid eye movement sleep, and reduction of mid-sleep awakenings, preferably two or more.

Reduced sleepiness upon waking means that the feeling of sleep deprivation upon waking is reduced.

Fatigue recovery upon waking up means that you feel less tired when you wake up.

A decrease in sleep onset time refers to a decrease in the time until falling asleep.

The increase in NREM sleep refers to the increase in the ratio of NREM sleep time to total sleep time.

Reduction in mid-sleep awakenings refers to a decrease in the number of awakenings during sleep.

[Preparation] In the present invention, the oral preparation of the present invention can be orally administered in the form of solid preparations such as tablets, capsules, granules, syrups, powders, etc. Solid preparations can use excipients, lubricants, binders, disintegrants; liquid preparations can use solvents, dissolution aids, suspending agents, tonic agents, buffers, analgesics, etc. In addition, additives such as preservatives, antioxidants, colorants, sweeteners, etc. can also be used as needed.

Examples of the excipient include: sugar alcohols such as D-sorbitol, mannitol, and xylitol; sugars such as glucose, white sugar, lactose, and fructose; crystalline cellulose, sodium carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose, calcium hydrogen phosphate, wheat starch, rice starch, corn starch, potato starch, dextrin, β-cyclodextrin, light anhydrous silicic acid, titanium oxide, magnesium aluminum silicate, talc, kaolin, olive oil, and the like.

Examples of the binder include cellulose derivatives such as methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, acrylic polymers, gelatin, gum arabic, polytriglucose, α-starch, agar, tragacanth gum, sodium alginate, propylene glycol alginate, and the like.

In one embodiment, as the shaping agent or the binder, at least one selected from the group consisting of dried powder of fresh leaves of evergreen daylily, fermented vinegar, dried powder of fermented vinegar, fermented glutinous rice and oligosaccharide cycloglucan (CI) can be used.

Examples of disintegrants include starch, low-substituted hydroxypropyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, hydroxypropyl starch, partially alpha-starched starch, and the like.

Examples of the solvent include water for injection, alcohol, propylene glycol, polyethylene glycol, sesame oil, corn oil, and the like.

Examples of lubricants include stearic acid, magnesium stearate, calcium stearate, polyoxyl stearate, cetyl alcohol, talc, hydrogenated oil, sucrose fatty acid ester, dimethyl polysiloxane, beeswax, white beeswax, and the like.

Examples of the dissolution aid include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, triaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate, and the like.

Examples of suspending agents and emulsifiers include: surfactants such as stearyl triethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzyl ammonium chloride, benzathonine chloride, and glyceryl monostearate; hydrophilic polymers such as polyvinyl alcohol, polyvinyl pyrrolidone, sodium carboxymethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose; and waxes such as wormwood wax, beeswax, carnauba wax, whale wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, cyclic lanolin, lanolin wax, candelilla wax, wood wax, lignite wax, wormwood wax, and rice bran wax.

As isotonic agents, for example, sodium chloride, glycerol, and D-mannitol can be mentioned.

Examples of the buffer include phosphate, acetate, carbonate, citrate and other buffer solutions.

Examples of preservatives include p-hydroxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, and the like.

As an implementation form, at least one selected from the group consisting of fermented vinegar, dried powder of fermented vinegar and fermented glutinous rice can be used as a substitute for the preservative.

Examples of antioxidants include sulfites, ascorbic acid, etc.

When the oral preparation of the present invention is made into a solid preparation, a manufacturing method known in the art can be used. For example, the following methods can be cited: kneading the oral preparation composition, passing it through a screen, thereby forming an extruded granulation, and crushing and sizing the formed extruded granulation; adding kneading water to the above composition, forming and stirring granulation by a vertical granulator, and then crushing and sieving using a sizing machine (Comil). Another method can be cited: after compressing the above preparation composition by a wheel press, crushing and sieving using a roller granulator. Another method can be cited in which the fluidized layer is dried after stirring and granulating. Furthermore, for example, in the case of direct tableting, after mixing the components, they can be directly put into a tableting machine for tableting.

As an embodiment of the oral preparation, the fermented extract of fresh leaves of daylily can be frozen to prepare a solid preparation. Alternatively, for example, the fermented extract of fresh leaves of daylily can be mixed with dried powder of fresh leaves of daylily and then frozen to prepare a solid preparation.

[Beverages] The oral preparation of the present invention can also be used as a beverage, and can be provided by being included in a food or functional food. Examples of such food or functional food include: rice; various noodles including buckwheat noodles, udon noodles, vermicelli, Chinese noodles, instant noodles, and cup noodles; soft drinks, carbonated drinks, nutritional drinks, fruit juice drinks, lactic acid drinks, sports drinks, etc.; curry syrup, stews, and various soups; cold drinks such as ice cream, slush, and Japanese shaved ice; candies, cookies, candies, chewing gum, chocolate, tablet candies (tablet candy), snacks, cookies, jelly, jam, cream, other baked cakes and other snacks; Japanese fish cakes, fish cakes, ham, sausages and other seafood and livestock processed foods; processed milk, fermented milk and other dairy products; salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, sauce and other fats and fat processed foods; sauce, seasoning, miso, soy sauce, sauce and other condiments; soups, stews, salads, home-cooked dishes, pancakes, pickles; other forms of health, nutritional supplements, functional labeled foods, specific health foods, etc.

Furthermore, supplements (powders, granules, soft capsules, hard capsules, tablets, chewable tablets, fast-disintegrating tablets, syrups, liquids, etc.) containing the present invention can also be prepared.

Furthermore, the composition of the present invention may be contained in food for animals such as pets.

Additives may be added to the food as needed. Examples of such additives include glucose, fructose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn syrup, lactose, mannitol, dextrin, citric acid, sodium citrate, tartaric acid, apple acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium isoscorate, glycerin, propylene glycol, glycerol fatty acid esters, polyglycerol fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, vitamin C, vitamin B group, vitamin E, niacinamide, calcium pantothenate, amino acids, calcium salts, surfactants, pigments, flavors, preservatives, and the like.

The present invention can be used for food and drink that are allowed to be labeled to improve or prevent various symptoms or conditions. Here, in the present invention, food and drink that are allowed to be labeled to improve or prevent various symptoms or conditions refers to food and drink with medicinal effects that are allowed and designated by the state or public groups, such as functional labeled food, specific health food and other health functional food, special purpose food, etc. Furthermore, although the names or regulations may change due to different situations, times, and systems of different countries, those that are essentially the same are included in the present invention.

In the present invention, the blending amount of the composition of the present invention is not particularly limited and can be appropriately set according to the applicable target (type of target disease or symptom, etc.); applicable part; gender or age of the user; the form of food, functional label food, specific health food, quasi-drug or drug; the method and frequency of administration or intake thereof; palatability, etc.

The composition of the present invention is not particularly limited in dosage form. For example, as an edible food composition, there can be exemplified functional foods such as supplements and nutritional drinks; snacks such as soft candies, candies, chewing gums, crackers, and cookies; foods such as ice cream, jelly, yokan, bread, chawanmushi, and curry; beverages such as milk drinks, refreshing water, and soups; and seasonings such as sauces and ponzu vinegar.

In addition, from the perspective of improving sleep quality as an effect of the present invention, the composition of the present invention is preferably prepared into a functional food for consumption. Functional foods include functional labeled foods, nutritional functional foods, nutritional supplementary foods, and specific health foods. In terms of being able to clearly label the use, functional labeled foods are preferred.

[Applicable subjects] The subjects to whom the composition of the present invention is applicable are not particularly limited, as long as they have a need to improve sleep quality. They can be people of an age group whose sleep time is likely to be short due to the need to study or work, such as those aged 15 or above, 20 or above, or 30 or above. They can also be people of an age group who are prone to physical changes due to changes in hormone balance, such as middle-aged people (approximately 45 years old or above and less than 55 years old) or elderly people (55 years old or above).

[pH] The pH of the composition of the present invention can be appropriately set according to the type and content of other blending ingredients, the form of the preparation, the method of use, etc., and is not particularly limited. It can be within the physiologically or pharmaceutically permissible range, for example, it can be set to pH 2 to 9. From the perspective of stably exerting the effect of the present invention, the pH of the composition of the present invention is preferably set to 2 to 9.5, more preferably 3 to 9, further preferably 4 to 8.5, further preferably 4.5 to 8, and further preferably 5.5 to 7.5.

[Implementation form] The above implementation form is not particularly limited, and the present invention discloses the following implementation forms.

An oral preparation for improving sleep quality contains a fermented extract of fresh leaves of evergreen daylily as an active ingredient.

For the oral preparation mentioned above, the sleep quality refers to the reduction of sleepiness upon waking up, the recovery of fatigue upon waking up, the reduction of the time to fall asleep, the increase of non-rapid eye movement sleep, or the reduction of awakening during the sleep.

For the oral preparation as mentioned above, the sleep quality refers to a reduction in the time it takes to fall asleep, an increase in non-rapid eye movement sleep, and/or a reduction in awakenings during sleep.

In the oral preparation, the water content of the fresh leaves of the evergreen daylily is 10% or more.

In the oral preparation mentioned above, the fresh leaves of the evergreen daylily are obtained within four months after the flowers of the evergreen daylily have fallen.

In the oral preparation mentioned above, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested one to four months after the flowers fall.

In the oral preparation mentioned above, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested three to four months after the flowers fall.

The oral preparation as described above, wherein the fermentation is produced by fermentation treatment using yeast.

In the oral preparation as mentioned above, the yeast is Saccharomyces cerevisiae or a subspecies thereof.

The oral preparation as described above, wherein a treatment to inactivate the enzyme contained in the fresh leaves of the evergreen daylily is performed before or after the fermentation treatment.

In the oral preparation as mentioned above, the treatment for inactivating the enzyme contained in the fresh leaves of the evergreen daylily is a heating treatment or a freezing treatment.

In the oral preparation as mentioned above, the temperature condition of the heat treatment is 60-100°C.

In the oral preparation as mentioned above, the heating treatment time condition is 10 seconds to 10 minutes.

The oral preparation as described above, wherein the temperature condition of the freezing treatment is below -4°C.

The oral preparation as described above, wherein the temperature condition of the freezing treatment is below -20°C.

In the oral preparation, the daily intake of the fresh leaf fermented extract of the evergreen daylily is 100 mg/day to 4000 mg/day in terms of dry solid content.

As for the oral preparation mentioned above, the content of oxo-jasmine in 100 g of the fresh leaf fermentation extract of evergreen daylily is 2 mg to 20,000 mg.

A use of a fermented extract of fresh leaves of evergreen daylily for preparing a sleep quality improving agent.

As in the above-mentioned uses, the above-mentioned sleep quality refers to the reduction of sleepiness when waking up, the recovery of fatigue when waking up, the reduction of the time to fall asleep, the increase of non-rapid eye movement sleep, or the reduction of awakening in the middle of the night.

As in the above-mentioned use, wherein the above-mentioned sleep quality refers to the reduction of the time to fall asleep, the increase of non-rapid eye movement sleep, and/or the reduction of awakenings during the sleep.

As for the above use, the moisture content of the fresh leaves of the evergreen daylily is greater than 10%.

As for the above-mentioned uses, the fresh leaves of the evergreen daylily are obtained within four months after the flowers fall.

As mentioned above, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested one to four months after the flowers fall.

As mentioned above, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested three to four months after the flowers fall.

The use as mentioned above, wherein the fermentation is produced by fermentation treatment using yeast.

The use as mentioned above, wherein the yeast is brewing yeast (Saccharomyces cerevisiae) or its subspecies.

The use as described above, wherein a treatment to inactivate the enzyme contained in the fresh leaves of the evergreen daylily is performed before or after the fermentation treatment.

In the above-mentioned use, the treatment for inactivating the enzyme contained in the fresh leaves of the evergreen daylily is a heating treatment or a freezing treatment.

As for the above-mentioned use, wherein the temperature condition of the above-mentioned heat treatment is 60-100°C.

For the above-mentioned use, the time condition of the above-mentioned heat treatment is 10 seconds to 10 minutes.

As for the above use, wherein the temperature condition of the above freezing treatment is below -4°C.

As for the above use, wherein the temperature condition of the above freezing treatment is below -20°C.

In the above-mentioned use, the daily intake of the fermented extract of fresh leaves of evergreen daylily is 100 mg/day to 4000 mg/day in terms of dry solid content.

As for the above use, the content of oxo-jasmine in 100 g of the fresh leaf fermentation extract of evergreen daylily is 2 mg to 20000 mg.

A fermented extract of fresh leaves of evergreen daylily is used for improving sleep quality.

As in the extract mentioned above, wherein the sleep quality refers to the reduction of sleepiness upon waking up, the recovery of fatigue upon waking up, the reduction of the time to fall asleep, the increase of non-rapid eye movement sleep, or the reduction of awakening during the sleep.

As in the extract above, wherein the sleep quality refers to a reduction in the time it takes to fall asleep, an increase in non-rapid eye movement sleep, and/or a reduction in awakenings during the night.

As mentioned above, the extract, wherein the water content of the fresh leaves of the evergreen daylily is greater than 10%.

As in the extract mentioned above, the fresh leaves of the evergreen daylily are obtained within four months after the flowers fall.

In the extract as mentioned above, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested one to four months after the flowers fall.

In the extract as mentioned above, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested three to four months after the flowers fall.

The extract as above, wherein the fermentation is produced by fermentation treatment using yeast.

The extract as mentioned above, wherein the yeast is brewing yeast (Saccharomyces cerevisiae) or its subspecies.

The extract as described above, wherein, before or after the fermentation treatment, a treatment is performed to inactivate the enzyme contained in the fresh leaves of the evergreen daylily.

As in the above-mentioned extract, the treatment for inactivating the enzyme contained in the fresh leaves of the above-mentioned evergreen daylily is heating treatment or freezing treatment.

The extract as described above, wherein the temperature condition of the above-mentioned heat treatment is 60-100°C.

The extract as described above, wherein the heating treatment time condition is 10 seconds to 10 minutes.

The extract as described above, wherein the temperature condition of the above freezing treatment is below -4°C.

The extract as described above, wherein the temperature condition of the above freezing treatment is below -20°C.

As for the extract mentioned above, the daily intake of the fermented extract of fresh leaves of evergreen daylily is 100 mg/day to 4000 mg/day calculated as dry solid content.

As for the extract mentioned above, the content of oxo-jasmine in 100 g of the fresh leaf fermentation extract of evergreen daylily is 2 mg to 20,000 mg.

A method for improving sleep quality comprises allowing a human to ingest a fermented extract of fresh leaves of evergreen daylily as an active ingredient.

In the above method, the sleep quality refers to the reduction of sleepiness upon waking up, the recovery of fatigue upon waking up, the reduction of the time to fall asleep, the increase of non-rapid eye movement sleep, or the reduction of awakening during the sleep.

In the method as described above, the sleep quality refers to a reduction in the time it takes to fall asleep, an increase in non-rapid eye movement sleep, and/or a reduction in awakenings during sleep.

As in the above method, wherein the moisture content of the fresh leaves of the evergreen daylily is greater than 10%.

In the above method, the fresh leaves of the evergreen daylily are obtained within four months after the flowers fall.

In the above method, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested one to four months after the flowers fall.

In the above method, the fresh leaves of the evergreen daylily are obtained from the evergreen daylily harvested three to four months after the flowers fall.

The method as described above, wherein the fermentation is produced by fermentation treatment using yeast.

The method as described above, wherein the yeast is brewing yeast (Saccharomyces cerevisiae) or a subspecies thereof.

As described above, before or after the fermentation treatment, a treatment is performed to inactivate the enzyme contained in the fresh leaves of the evergreen daylily.

In the above method, the treatment for inactivating the enzyme contained in the fresh leaves of the evergreen daylily is a heating treatment or a freezing treatment.

As in the above method, the temperature condition of the above heat treatment is 60-100°C.

In the above method, the heating treatment time is between 10 seconds and 10 minutes.

As in the above method, wherein the temperature condition of the above freezing treatment is below -4°C.

As in the above method, wherein the temperature condition of the above freezing treatment is below -20°C.

In the above method, the daily intake of the fermented extract of fresh leaves of evergreen daylily is 100 mg/day to 4000 mg/day in terms of dry solid content.

As in the above method, the content of oxo-jasmine in 100 g of the fresh leaf fermentation extract of the evergreen daylily is 2 mg to 20,000 mg. [Example]

Next, the present invention is specifically described by using embodiments and test examples, but the present invention is not limited to the following embodiments and test examples.

[Preparation of fermented extract of fresh leaves of evergreen daylily] The fresh leaves of evergreen daylily used are those produced in Okinawa Prefecture, Japan. Specifically, fresh leaves of evergreen daylily cultivated in Okinawa Prefecture, Japan, were obtained about two months after the flowers fell.

1 kg of fresh leaves of daylily are cut into about 6 cm lengths, crushed and squeezed, and purified by centrifugal separation to obtain juice. Yeast (Saccharomyces cerevisiae) at a concentration of 0.5-1% is added to the juice concentrated to a Brix value of about 25%, and fermented at 35℃-38℃ for about 3 hours. The fermented product is heated to 90℃ to inactivate the enzyme and then cooled to obtain a fermented extract of fresh leaves of daylily. When storing the fermented extract of fresh leaves of daylily, it is frozen at a temperature below -18℃ to inhibit the decomposition of oxo-erucic acid.

In another embodiment, 1 kg of fresh leaves of daylily are cut into about 6 cm lengths, crushed and ground to make a paste. Yeast (Saccharomyces cerevisiae) at a concentration of 0.5-1% is added to the paste, and fermentation is carried out at 35°C-38°C for about 3 hours. The fermented product is heated to 90°C to inactivate the enzyme and then cooled to obtain a fermentation extract of fresh leaves of daylily. When storing the fermentation extract of fresh leaves of daylily, it is frozen at a temperature below -18°C to inhibit the decomposition of oxo-erucic acid. In another embodiment, 1 kg of fresh leaves of the evergreen daylily are not cut but fermented, thereby processing them into a paste.

The inventors of the present invention have found that the oxo-galactoenamine contained in the fresh leaves of the evergreen daylily will be decomposed and reduced due to the presence of sugars, and that an extract significantly containing oxo-galactoenamine can be obtained by fermenting the fresh leaves of the evergreen daylily. In addition, it has been found that the oxo-galactoenamine will be decomposed and reduced due to the presence of various enzymes contained in the fresh leaves of the evergreen daylily, and a method of inactivating various enzymes by heat treatment is adopted. It is confirmed that by using these methods to produce, in 100 g of the fermented extract of the fresh leaves of the evergreen daylily, the oxo-galactoenamine content is high, at 2 mg to 2000 mg.

[Test Example 1. Evaluation of Clinical Trial] In order to study the sleep-improving effect of the fermented extract of fresh leaves of daylily, a randomized placebo-controlled parallel group trial was conducted on 32 healthy subjects with sleep disturbance. As a placebo food, dextrin was used instead of the fermented extract of fresh leaves of daylily.

The subjects were healthy men and women aged 20 to 65, mainly those who worked in clerical jobs and had mild sleep disturbances such as feeling easily tired, having difficulty falling asleep, and having shallow sleep, but were not suffering from any disease.

(Test Method) This test was conducted as a randomized double-blind placebo-controlled parallel group test. As a screening test, 40 subjects were asked to come to the hospital for PSQI sleep questionnaire survey, blood test, urine test and interview, and 32 subjects who met the selection criteria of this test and did not meet the exclusion criteria were selected. As a pre-intake examination, the selected subjects recorded the OSA sleep questionnaire survey when they woke up for one week, and wore a sleep electroencephalogram (ZA-1, Proassist Ltd.) to measure sleep electroencephalograms one day before work. After the pre-intake examination, the selected subjects were asked to consume the test food four capsules a day for two weeks. During the sampling period, the selected subjects were asked to fill out the OSA sleep questionnaire when they woke up every day, and record their physical condition in a diary every day. In addition, as in the pre-sampling examination, they wore a sleep electroencephalogram device to measure their sleep electroencephalograms one day before the working day of the second week of the sampling. After the sampling period, the selected subjects came to the hospital again for blood and urine tests, and a post-sampling examination was carried out, that is, the PSQI sleep questionnaire and interview were conducted.

Regarding the allocation of the test drinks, the allocation manager who did not participate in the implementation of this trial randomly allocated them using the permuted block method, which used age, gender, and PSQI as stratification factors. 16 subjects were assigned to each of the test product group or the placebo food group, and the intake began. The intake period was set to two weeks. During the intake period, the sleep quality survey was also implemented using the OSA sleep questionnaire, and sleep EEG was measured in the second week of intake. All subjects completed the trial and no one dropped out. However, one person in the placebo intake group failed to obtain the sleep EEG data, so it was considered blank.

The OSA sleep questionnaire is a psychological scale that evaluates sleep introspection when waking up, and is also used in sleep clinics to determine sleep quality. The sleep questionnaire is divided into 5 factors.

Regarding the results of blood and urine tests, no abnormal changes were found in the two groups before and after the intake. In addition, no adverse events with a causal relationship with the intake of the tested food were observed.

In this test, the results are expressed as mean ± standard deviation. The significance level of the test was set at 5% on both sides, and the inter-group comparison was performed by Student's t-test, and the intra-group comparison was performed by a corresponding t-test. Statistical analysis was performed using SAS Ver.9.4 (manufactured by SAS Corporation). The results are shown in Figures 1 and 2. The "*" in Figures 1 and 2 indicates that there is a significant difference when p < 0.05.

(Results of OSA sleep questionnaire) As shown in Figures 1 and 2, in the analysis of the OSA sleep questionnaire, in the group taking oxo-ergine from daylily (referred to as forget-me-not in each figure) compared with the placebo, significant differences were observed in the I factor "sleepiness upon waking up" and the IV factor "fatigue recovery", and sleep quality was improved by taking oxo-ergine from daylily.

(Results obtained using a sleep electroencephalogram) Sleep was evaluated by measuring the brain waves of the subjects when they went to bed. Sleep quality was evaluated based on the delta power value, which represents the amount of low-frequency brain waves (delta waves) that appear during non-rapid eye movement sleep. The amount of non-rapid eye movement sleep was evaluated based on the duration of non-rapid eye movement sleep during sleep. Sleep rhythm was evaluated based on the duration of the initial sleep cycle (falling asleep-non-rapid eye movement sleep-rapid eye movement sleep). The results are shown in Figure 3.

As shown in Figure 3, in the sleep electroencephalogram analysis, the ratio of mid-wake time, the change in sleep onset time after two weeks of intake, and the change compared with before intake in the group taking daylily were significantly reduced compared with the group taking placebo. In addition, the ratio of non-rapid eye movement sleep in the group taking daylily was significantly increased compared with the group taking placebo.

The change in the rate of rapid eye movement sleep (%) of the daylily intake group compared with before intake was significantly reduced compared with the placebo group.

[Test Example 2. Evaluation of seasonal changes in oxo-opiotinamide in various parts of evergreen daylilies] In order to effectively utilize medicinal plants such as evergreen daylilies for food, it is believed that it is best to pick and use them when they contain more active ingredients. Therefore, it is believed that it is necessary to study the temporal changes of active ingredients during plant growth, and the changes of oxo-opiotinamide (denoted as OPT in each figure) in evergreen daylilies were studied.

The flowers, leaves, and roots of the evergreen daylily were collected in a fresh state, left at room temperature overnight without drying, extracted with methanol, and then concentrated under reduced pressure to prepare an extract.

Dissolve 10 mg of the obtained extract in purified water to make exactly 5 ml. Adsorb 2.5 ml of it on a Sep-PAC C18 cartridge and dissolve it with 2 ml of purified water. Add purified water to the dissolution solution to make the total volume exactly 5 ml as the test solution, and perform OPT detection by fluorescence detection using HPLC. In addition, the quantitative value is converted into the amount of OPT per 1 g of fresh sample (n=3).

(HPLC conditions) Pump: Shimadzu LC-10AS Column: CAPCELL PAK C ₁₈ (UG120), 150×4.6 mm, 5 μm Elution: 10 mM CH ₃ COONa, 0.2 mM (CH 3 COO) ₂ Cu, 0.8 mM CH ₃ (CH ₂ ) ₇ SO _{3 Na} , 1.0% CH ₃ CN pH 5.6 adjusted by _{CH 3 COOH} Flow rate: 1.0 ml/min Detector: JASCO 821-FP (λ _ex 350 nm, λ _em 460 nm) Reaction coil: 3 m×0.5 mm (ID), 55°C Pump: JASCO 880-PU Reaction reagent Reagent): 0.3 MH ₃ BO ₃ , 3.0 mg/l 2-hydroxyethanol, 40 mg/l phthalaldehyde pH 10.5 adjusted by NaOH solv. Flow rate: 0.5 ml/min

The results of evaluating the seasonal changes of oxo-erucic acid accompanying flowering are shown in Figure 4. In addition, the results of evaluating the changes of oxo-erucic acid in leaves and roots before flowering are shown in Figure 5.

As shown in Figure 4, the content of oxo-jasmine in fresh flowers is higher in the early bud stage, and shows a decreasing trend as the flowers bloom.

As shown in Figure 5, the oxo-juniorhinine content in fresh leaves and roots showed a constant value until just before flowering, and an increase was confirmed after flowering (after the flowering period). In Figure 5, the oxo-juniorhinine content increased significantly within one month after the flowering period and within one month after the flowers fell (after the flowering period).

Since oxo-juniorhines are relatively stable when dissolved in water or at pH 1.2 to pH 6.8, it is suggested that oxo-juniorhines may be accumulated at a high density in the early stage of plant organs and consumed during the growth process. Therefore, it is confirmed that the fresh leaves of the evergreen daylily are preferably collected within one month after the flowers fall and are prepared into extracts without drying, but it is not limited to this. In this case, in order to inhibit the decomposition and reduction of oxo-juniorhines contained in the fresh leaves of the evergreen daylily due to the presence of sugars, it is more preferable to use yeast, such as yeast, to perform fermentation treatment.

[Test Example 3. 28-day repeated oral toxicity test of the fermented extract of fresh leaves of evergreen daylily in rats] The extract prepared by the above method was powdered by conventional methods and orally administered to five male and female rats at the maximum dosage of 1000 mg/kg specified in the guidelines for 28 days to check for toxicity.

The results showed that in the hematological examination, the activated partial thromboplastin time and prothrombin time in the test substance group of male rats were significantly prolonged compared with the control group. However, such changes were not observed in the female rat group. No bleeding tendency was observed in the subcutaneous findings of the test substance group of male rats during autopsy, and no blood biochemistry and pathological changes showing liver abnormalities were found. It is difficult to believe that the synthesis of various factors of the extensive coagulation system involving the endogenous coagulation system and the exogenous coagulation system was hindered. Therefore, it is believed that the significant prolongation of the activated partial thromboplastin time and prothrombin time observed in the test substance group of male rats is not a direct toxic reaction caused by the administration of the test substance.

Myocardial degeneration/fibrosis was observed in one male rat in the test substance administration group. This change was observed in the test substance administration group, but since the change was extremely mild and no other animals in the same group had the same change, and since the aspartate aminotransferase activity (AST) value in the blood biochemistry test of the above case was not increased, it was considered that the myocardial degeneration/fibrosis of this animal was an incidental finding.

The results of organ weight measurement showed that significantly low values of the absolute and relative weights of the heart were observed, but the degree of weight reduction was extremely low. In the histopathological examination of the heart, the above-mentioned extremely mild myocardial degeneration/fibrosis was observed in one case, but the heart weight of this animal was higher than that of other animals in the same group, so it is obvious that myocardial degeneration/fibrosis is not related to the reduction in heart weight. Apart from this finding, no abnormal histopathological findings were found, so the reduction in heart weight is not considered to be an organic change. On the other hand, the absolute weight of the lungs was also significantly reduced compared with the control group, but no abnormal histopathological findings were observed in the lungs, so the weight reduction is not considered to be a toxic change.

In addition, no abnormalities were observed in the general state, and no changes that could be attributed to the administration of the test substance were observed in the changes in body weight, food intake, ophthalmological examinations, urine tests, and blood biochemistry tests.

Based on the above, when the extract powder was repeatedly administered at a dosage of 1000 mg/kg for 28 days, it was determined that it did not cause toxicity to rats.

without

[Figure 1] is a graph showing the results of the OSA sleep questionnaire survey (factor I: sleepiness when waking up) in Test Example 1. [Figure 2] is a graph showing the results of the OSA sleep questionnaire survey (factor IV: fatigue recovery) in Test Example 1. [Figure 3] is a graph showing the results of the sleep electroencephalogram (sleep onset time, ratio of mid-waking time, non-rapid eye movement sleep) in Test Example 1. [Figure 4] is a graph showing the results of the evaluation of the seasonal changes in oxypinnatanine contained in the flowers of the evergreen daylily in Test Example 2. [Figure 5] is a graph showing the results of the evaluation of the seasonal changes in oxypinnatanine contained in the leaves or roots of the evergreen daylily in Test Example 2.

Claims (5)

A use of a fermented extract of fresh leaves of Hemerocallis fulva var. sempervirens for preparing an oral preparation for reducing sleepiness when waking up, relieving fatigue when waking up, reducing the time of falling asleep, or reducing awakening during the night, wherein the fermented extract of fresh leaves of Hemerocallis fulva var. sempervirens is obtained by fermentation using sugar-decomposing bacteria. The use of claim 1, wherein the sugar-degrading bacteria is yeast. For example, the use of claim 1, wherein the moisture content of the fresh leaves of the evergreen daylily is greater than 10%. For use as in claim 1, wherein the fresh leaves of the evergreen daylily are obtained within four months after the flowers of the evergreen daylily have fallen. For example, in the use of claim 1, the daily intake of the fermented extract of fresh leaves of evergreen daylily is 100 mg/day to 4000 mg/day calculated as dry solid content.