WO2021112176A1

WO2021112176A1 - Composition for improving quality of sleep

Info

Publication number: WO2021112176A1
Application number: PCT/JP2020/045036
Authority: WO
Inventors: 哲渡嘉敷; 案理酒井; 竜嗣竹田
Original assignee: 株式会社クレイ沖縄; 哲渡嘉敷; 渡嘉敷朝子
Priority date: 2019-12-03
Filing date: 2020-12-03
Publication date: 2021-06-10
Also published as: CN115023234A; TW202133873A; JPWO2021112176A1

Abstract

Provided is an oral preparation which can exert a sleep quality improving effect effectively.　An oral preparation for improving the quality of sleep is prepared, which contains a fermentation extract of fresh leaves of orange daylily (Hemerocallis fulva L. var. sempervirens) as an active ingredient.

Description

Composition for improving sleep quality

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

Approximately one-third of life is occupied by "sleep", and it is important to improve the quality of sleep. Data support that sleep is also closely related to the rhythm and stress of life. For example, according to the "2000 Insurance and Welfare Trend Survey" released by the Ministry of Health, Labor and Welfare, it was pointed out that the higher the stress, the more likely it is that problems such as "no feeling of deep sleep even when waking up in the morning" and "difficulty falling asleep" are likely to occur. ing.

Development has been made to improve sleep quality with plant-derived components. For example, Patent Document 1 proposes a sleep quality improving agent containing an extract of Ash genus plant or Siberian larix sibir.

Akinowasuregusa (Okinawa name: Kwansou, Niibui grass), which is a plant belonging to the genus Daylily of the Liliaceae family, is said to be effective for sleeplessness since the Ryukyu dynasty. It has been eaten as a kind of medicinal food. In modern times, it is also used for beverages such as tea and confectionery, and flowers are beautiful for ornamental purposes and are attracting attention as a representative material of Okinawa.

International Publication No. 2013/05/1727

Although it is known that daylily (daylily) contains a compound that has an effect on sleep improvement, when prepared as an extract of daylily (daylily), the compound that has an effect on sleep improvement is efficient. The technology for good inclusion is underdeveloped.

In addition, no detailed tests have been conducted on what kind of sleep-improving effect can be expected for humans when prepared as an daylily extract. There are many different sleep qualities, including sleep improvement, such as shortening sleep onset time, increasing total non-REM sleep time leading to deep sleep, depth of non-REM sleep, reduced mid-wake awakening, ease of waking up, and waking up. There are various needs for those who are worried about the quality of sleep, such as reduction of sleepiness during time and improvement of fatigue after waking up. That is, no clinical trials have been conducted on what kind of sleep quality improvement can be approached to humans when prepared as an daylily extract.

In view of the above problems, as a result of diligent studies, it was found that an oral preparation having a remarkable effect on improving the quality of sleep can be obtained by preparing a fermented extract using fresh leaves without drying the daylily. By conducting clinical trials on the fermented leaf extract of daylily daylily in humans, it was found that it has a significant effect on the reduction of sleep onset time and the increase of deep sleep, among other sleep qualities, and completed the present invention. ..

That is, the present invention provides the compositions listed below.
Item 1.
An oral preparation for improving sleep quality that contains fermented fresh leaf extract of daylily as an active ingredient.

Item 2.
Item 2. The oral preparation according to Item 1, wherein the quality of sleep is reduction of drowsiness at the time of waking up, recovery from fatigue at the time of waking up, reduction of sleep onset time, increase of non-REM sleep, or reduction of awakening during sleep.

Item 3.
Item 2. The oral preparation according to

Item

1 or 2, wherein the water content of the fresh leaves of the daylily is 10% or more.

Item 4.
The oral preparation according to any one of Items 1 to 3, wherein the fresh leaves of the daylily are obtained from the daylily within 4 months after the flower fall.

Item 5.
The oral preparation according to any one of Items 1 to 4, wherein the daily intake of the fermented fresh leaf extract of Akinowasuregusa is 100 mg / day to 4000 mg / day in terms of dry solid content.

It is possible to improve the quality of sleep by applying an oral preparation containing a fermented leaf extract of Akinowasuregusa as an active ingredient.

FIG. 1 is a graph showing the results of the OSA sleep questionnaire (factor I: drowsiness when waking up) in Test Example 1. FIG. 2 is a graph showing the results of the OSA sleep questionnaire (factor IV: recovery from fatigue) in Test Example 1. FIG. 3 is a graph showing the results of a sleep electroencephalograph (sleep onset latency, halfway awakening time ratio, non-REM sleep) in Test Example 1. FIG. 4 is a graph showing the results of evaluating the seasonal variation of oxypinnatanin contained in the flowers of the daylily daylily in Test Example 2. FIG. 5 is a graph showing the results of evaluating the seasonal variation of oxypinnatanin contained in the leaves or roots of Daylily chinensis in Test Example 2.

[Fermented fresh leaf extract of daylily]
Daylily (Hemerocallis fulva var. Semipervirens) is a plant belonging to the genus Daylily of the Liliaceae family. "Akinowasuregusa" is a Japanese name, and in Okinawa Prefecture, it is also called by dialect names such as Kwansou, Kwansou, and Nibuigusa.

The production area of the daylily is not particularly limited, but for example, domestic or foreign production can be mentioned, and if it is domestic, for example, Okinawa Prefecture, southern Kyushu, Nansei Islands, etc. can be mentioned, and if it is foreign, for example, Asia such as China and Taiwan can be mentioned.

In the oral preparation of the present invention, from the viewpoint of remarkably improving the quality of sleep, the site from which the daylily extract is extracted is from the whole plant or leaves, stems, buds, roots, and flowers. Although it is possible to use at least one selected from the above group, it is preferable to use at least the leaves of Daylily daylily.

The present inventors have newly found that the content of oxypinnatanine in the leaves of daylily is significantly reduced by undergoing the drying process. Therefore, the leaves of the daylily daylily are preferably fresh leaves (undried).

In one embodiment, it is also possible to use a stem and / or a bud of the daylily as the extraction site in combination with the leaf of the daylily. Even when these extraction sites are used, it is preferable to use them in a raw state because the content of oxypinnatanine is significantly reduced by the drying step.

In one embodiment, the present inventors have found that the content of oxypinnatanine is significantly reduced by flowering from the state of buds of the daylily flower, and the flower of the daylily is extracted. It is also preferable to exclude from.

Akinowasuregusa is preferably obtained by fermenting fresh leaves from the viewpoint of significantly improving the quality of sleep.

The water content of the fresh leaves (undried matter) of the daylily can be, for example, 10% or more, preferably 15% or more, more preferably 20% or more, still more preferably 25% or more. The above-mentioned water content is preferably the above-mentioned water content at the time of sale if it is a commercially available product of fresh leaves of Akinowasuregusa, and more preferably the above-mentioned water content at the time of being subjected to fermentation treatment.

The present inventors have newly found that the content of oxypinnatanine in the fresh leaves of daylily is significantly reduced depending on the harvest time. Specifically, it can be seen that the content of oxypinnatanine in the leaves increases remarkably 3 to 4 months after the flowering period has passed and the content of oxypinnatanin in the leaves decreases sharply 4 months after the flowering. It was issued. Therefore, the fresh leaves of Daylily are preferably harvested within at least 4 months after the fall of flowers, more preferably 1 to 4 months after the fall of flowers, and 2 to 4 months after the fall of flowers. It is more preferable that the daylily is harvested in a month, and it is particularly preferable that the daylily is harvested 3 to 4 months after the flower falls.

Fermentation means known to those skilled in the art may be used in the fermentation process for obtaining the fermented product. Examples of the fermentation means include aspergillus, lactic acid bacteria, and yeast. As the fermentation means, yeast, for example, yeast can be preferably used. By performing the fermentation treatment, it is possible to prevent the oxypinnatanine contained in the fresh leaves of the daylily daylily from being decomposed and reduced due to the presence of sugars.

Examples of Aspergillus include filamentous fungi belonging to the genus Aspergillus.

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

Examples of yeast include those belonging to the genus Saccharomyces, the genus Schizosaccharomyces, the genus Kluyveromyces, or the genus Pichia. As the yeast, Saccharomyces cerevisiae, which is also called yeast, or a subspecies thereof is preferable from the viewpoint of remarkably exerting the effect of the present invention.

Fermentation treatment conditions can be appropriately selected by those skilled in the art, for example. The conditions of the fermentation treatment include, for example, a step of mixing the yeast Saccharomyces cerevisiae with at least a part of the fresh leaves of the daylily, and it is preferable to mix it with the juice of the fresh leaves of the daylily.

The temperature condition of the fermentation treatment can 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 can be, for example, 20 to 70 hours, preferably 25 to 65 hours, and more preferably 30 to 60 hours.

After the fermentation treatment, a sterilization step may be included as appropriate.

The fermentation treatment may include, for example, a step of immersing at least a part of the fresh leaves of the daylily in water. It is possible to add the above-mentioned fermentation means, for example, yeast, to the fresh leaves of the daylily soaked in water, and to carry out the fermentation treatment.

The immersion time in the immersion step can be, for example, 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 dipping step may include a step of boiling water.

The fresh leaves of the daylily, which is applied to the fermentation treatment, may be crushed before being subjected to the fermentation treatment.

The fresh leaves of the daylily, which is applied to the fermentation treatment, may be squeezed before being subjected to the fermentation treatment. Without limitation, it is preferable to apply the juice of fresh leaves of the pressed daylily daylily to the fermentation treatment.

The fresh leaves of the daylily applied to the fermentation treatment may be ground or ground before the fresh leaves of the daylily are fermented. By applying the paste of fresh leaves of the daylily ground daylily to the fermentation treatment, the fermentation efficiency is increased, and the paste is suitable for, for example, solid formulation.

Although not limited, the fresh leaves of the daylily are heat-treated to inactivate various enzymes contained in the fresh leaves of the daylily before or after the fermentation treatment, preferably after the fermentation treatment. It is preferable to carry out. The temperature condition of the heat treatment can be, for example, 60 to 100 ° C., preferably 70 to 95 ° C., and more preferably 75 to 90 ° C. The time of the heat treatment can 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 daylily, but not limited to, are added to or in place of the heat treatment of the daylily before or after being subjected to the fermentation treatment, preferably after being subjected to the fermentation treatment. It is preferable to carry out a freezing treatment to inactivate various enzymes contained in the fresh leaves. As the temperature condition of the freezing treatment, for example, it is preferably -4 ° C or lower, more preferably -10 ° C or lower, further preferably -15 ° C or lower, and more preferably -18 ° C or lower. It is particularly preferable, and most preferably it is -20 ° C or lower.

Although not limited, the freezing treatment makes it possible to use the fermented leaf extract of Daylily daylily in frozen foods, frozen desserts, etc., and by inactivating various enzymes contained in the fresh leaf of Daylily daylily, oxypinnatanine It becomes possible to produce foods in which decomposition is suppressed.

As the oral preparation of the present invention, a fermented extract obtained by subjecting the fresh leaves of Akinowasuregusa to the above-mentioned fermentation treatment may be used, or a commercially available fermented product may be used.

In the oral preparation of the present invention, the daily intake of the fermented leaf extract of Daylily daylily can be, for example, 100 mg / day to 4000 mg / day, preferably 200 mg / day to 3500 mg / day in terms of dry solid content. Can be a day.

The oral preparation of the present invention may have an oxypinnatanine content of, for example, 2 mg / day to 20000 mg / day, preferably 2 mg / day to 10000 mg / day, and more preferably 2 mg / day to 2500 mg / day. Particularly preferably, it can be 2 mg / day to 2000 mg / day.

According to the present invention, it has been found that the extraction efficiency of components effective for sleep quality can be increased by using the fermented extract obtained by performing the above fermentation treatment on the fresh leaves of Daylily daylily. In one embodiment, 100 g of the fermented leaf extract of Daylily daylily preferably contains 2 mg or more of oxypinnatanine, for example, 2 mg to 20000 mg, 2 mg to 10000 mg, 2 mg to 2500 mg, 2 mg to 2000 mg, and the like. can do.

[Use]
The oral preparation of the present invention is used for improving the quality of sleep. As used herein, the term "improvement" refers to an improvement in the state of sleep quality, prevention or delay of deterioration of the state of sleep quality.

The quality of sleep may be the quality of sleep during the entire sleep period or the quality of sleep during a part of the sleep period. Sleep is preferably sleep in the first cycle, which allows the brain to rest efficiently. There are REM sleep and non-REM sleep in sleep, and REM sleep and non-REM sleep alternate during sleep. The sleep cycle is the period from the end of one REM sleep to the end of the next REM sleep. The first sleep cycle means the first sleep cycle after falling asleep.

The sleep quality refers to the observation of one or more, preferably two or more, of the reduction of drowsiness when waking up, the recovery of fatigue when waking up, the decrease in sleep onset time, the increase in non-REM sleep, and the decrease in awakening during sleep. ..

Reducing drowsiness when waking up means reducing the feeling of lack of sleep when waking up.

Fatigue recovery when waking up means that the feeling of fatigue when waking up is reduced.

A decrease in sleep onset time means that the time to fall asleep is shortened.

An increase in non-REM sleep means that the ratio of non-REM sleep time to the total sleep time increases.

A decrease in awakening means a decrease in the number of times you wake up in the middle of sleep.

[Formulation]
In the present invention, the oral preparation of the present invention can also be orally administered as a solid preparation such as tablets, capsules, granules, syrups and powders. Excipients, lubricants, binders, disintegrants for solid preparations; solvents, solubilizers, suspending agents, tonicity agents, buffers, soothing agents, etc. should be used for liquid preparations. Can be done. If necessary, additives such as preservatives, antioxidants, colorants, and sweeteners can also be used.

Excipients include, for example, sugar alcohols such as D-sorbitol, mannitol and xylitol, sugars such as glucose, sucrose, lactose and fructose, crystalline cellulose, carmellose sodium, croscarmellose sodium, calcium hydrogen phosphate and wheat starch. , Rice starch, corn starch, potato starch, dextrin, β-cyclodextrin, light anhydrous silicic acid, titanium oxide, magnesium aluminometasilicate, talc, kaolin, olive oil and the like.

Examples of the binder include cellulose derivatives such as methyl cellulose, ethyl cellulose, hydroxypropyl cellulose and hydroxypropyl methyl cellulose, polyvinylpyrrolidone, polyvinyl alcohol, acrylate-based polymers, gelatin, arabic gum, purulan, pregelatinized starch, canten, tragant, and the like. Examples thereof include sodium alginate and propylene glycol alginate.

In one embodiment, the excipient or binder is at least selected from the group consisting of dried moromi vinegar powder, mashed vinegar, dried mashed vinegar powder, mashed lees, and oligosaccharide cyclodextran (CI). It is also possible to use one type.

Examples of the disintegrant include starch, low-degree-of-substitution hydroxypropyl cellulose, carboxymethyl cellulose calcium, croscarmellose sodium, hydroxypropyl starch, and partially pregelatinized starch.

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

Examples of the lubricant include stearic acid, magnesium stearate, calcium stearate, polyoxyl stearate, cetanol, talc, hydrogenated oil, sucrose fatty acid ester, dimethylpolysiloxane, beeswax, and salashimitsuro.

Examples of the solubilizing agent include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like.

Examples of the suspending agent / emulsifier include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glycerin monostearate; for example, polyvinyl alcohol and polyvinyl. Hydrophilic polymers such as pyrrolidone, sodium carboxymethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose; for example, shelac wax, beeswax, carnauba wax, whale wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, cyclic lanolin, lanolin. Examples thereof include waxes such as wax, candelilla wax, mokuro, montan wax, cellac wax, and rice wax.

Examples of the tonicity agent include sodium chloride, glycerin, D-mannitol and the like.

Examples of the buffer include a buffer solution such as phosphate, acetate, carbonate, and citrate.

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

As one embodiment, at least one selected from the group consisting of moromi vinegar, dried powder of moromi vinegar, and mash lees can be used as a substitute component of a preservative.

Examples of the antioxidant include sulfites, ascorbic acid and the like.

When the oral preparation of the present invention is a solid preparation, a production method known in the art can be used. For example, a method of crushing and sizing an extruded granulated product which is formed by kneading an oral pharmaceutical composition and passing it through a screen, adding kneading water to the composition, and stirring the mixture by a vertical granulator. After granulation, a method of crushing and sieving using a combil can be mentioned. Further, a method in which the pharmaceutical composition is compressed with a roller compactor, then pulverized with a roll granulator and sieved is mentioned. Further, a method of drying the fluidized bed after stirring and granulating can be mentioned. Further, for example, in the case of producing by direct striking, the composition may be mixed and then directly put into a locking machine to lock.

As one embodiment of the oral preparation, it is also possible to freeze the fermented leaf extract of Akinowasuregusa to make a solid preparation. Further, for example, it is also possible to mix the fermented fresh leaf extract of Akinowasuregusa and the dried fresh leaf powder of Akinowasuregusa and freeze them to form a solid preparation.

[Food and drink]
The oral preparation of the present invention can also be used as a food or drink, and can be provided by being contained in a food or a functional food. Such foods or functional foods include rice; various noodles including buckwheat, udon, harusame, Chinese noodles, instant noodles, cup noodles; soft drinks, carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, sports drinks. Beverages such as: Curry roux, stew, various soups; Cold confectionery such as ice cream, ice sherbet, shaved ice; Candy, cookies, candies, gum, chocolate, tablets, snacks, biscuits, jelly, jam, cream, and other baked goods Confectionery such as; fishery and livestock processed foods such as kamaboko, hampen, ham, sausage; dairy products such as processed milk and fermented milk; Processed foods; seasonings such as sauces, dressings, miso, soy sauce, sauces; soups, stews, salads, prepared foods, sprinkles, pickles; other various forms of health / nutritional supplements / functional foods / foods for specified health use, etc. Is illustrated.

Further, supplements containing the present invention (powder, granule, soft capsule, hard capsule, tablet, chewable tablet, quick-disintegrating tablet, syrup, liquid preparation, etc.) may be prepared.

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

Additives are added to food and drink as needed. Such additives include, for example, glucose, fructose, sucrose, maltose, sorbitol, stebioside, rubusoside, corn syrup, lactate, mannitol, dextrin, citric acid, sodium citrate, tartrate, malic acid, succinic acid, Lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbicate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, arabic gum, carrageenan, casein, gelatin, pectin , Agar, vitamin C, vitamin Bs, vitamin E, nicotinic acid amide, calcium pantothenate, amino acids, calcium salts, surfactants, pigments, fragrances, preservatives and the like.

The present invention can be used for foods and drinks for which labeling of improvement / prevention of various symptoms and conditions is permitted. Here, in the present invention, the foods and drinks for which the labeling of improvement / prevention of symptoms and conditions is permitted are foods and drinks having medicinal effects permitted and designated by the national government and public organizations, for example. Foods with functional claims, foods with health claims such as specified health foods, foods for special purposes, etc. Although the names and regulations change depending on the situation, the times, and the systems of each country, 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 the purpose of application (target disease, type of symptom, etc.), target site, gender and age of the applicator, food and drink, food with functional claims, etc. , Specified health food, quasi-drug, or drug form, administration or ingestion method and frequency of these, preference, etc. are appropriately set.

The composition of the present invention is not particularly limited in terms of formulation and dosage form. For example, functional foods such as supplements and nutritional drinks, sweets such as gummies, candy, gum, crackers and biscuits, foods such as ice cream, jelly, yokanpan, bread, steamed tea, curry, milk drinks, soft drinks, etc. Beverages such as soups, dressings, and seasonings such as ponzu vinegar are exemplified as food and drink compositions that can be eaten and drink.

Further, the composition of the present invention is preferably taken as a functional food from the viewpoint of the effect of the present invention of improving the quality of sleep, and among the functional foods, a food with a functional claim, a food with a nutritional function, and a nutrition. Supplements and foods for specified health use can be mentioned, but foods with functional claims are preferable because their uses can be clearly labeled.

[Applicable person]
The subject to which the composition of the present invention is applied is not particularly limited as long as there is a need to improve the quality of sleep, but is 15 years old, who tends to have a short sleep time due to the need for study and work. It may be over 20 years old, over 30 years old, or over 30 years old. It may be a middle-aged person (around 45 years old or older and less than 55 years old) or an elderly person (55 years old or older) who is likely to feel a change in the body due to a change in hormonal balance or the like.

[PH]
The pH of the composition of the present invention is appropriately set according to the type and content of other compounding ingredients, the form of the preparation, the method of use, etc., and is not limited as long as it is within a physiologically or pharmaceutically acceptable range. The pH can be 2-9. From the viewpoint of stably exhibiting the effects of the present invention, the pH of the composition of the present invention is preferably 2 to 9.5, more preferably 3 to 9, still more preferably 4 to 8.5, and even more preferably. Can be 4.5 to 8, and even more preferably 5.5 to 7.5.

[Embodiment]
The present invention discloses the following embodiments with respect to the above-described embodiments, without limitation.

An oral preparation for improving sleep quality that contains fermented fresh leaf extract of daylily as an active ingredient.

The above-mentioned oral preparation whose sleep quality is reduction of drowsiness when waking up, recovery from fatigue when waking up, reduction of sleep onset time, increase of non-REM sleep, or reduction of awakening during sleep.

The above-mentioned oral preparation whose sleep quality is a decrease in sleep onset time, an increase in non-REM sleep, and / or a decrease in awakening.

The above-mentioned oral preparation in which the water content of the fresh leaves of the daylily Akinowasu is 10% or more.

The above-mentioned oral preparation in which the fresh leaves of the daylily are obtained from the daylily within 4 months after the flower falls.

The above-mentioned oral preparation in which the fresh leaves of the daylily are obtained from the daylily harvested 1 to 4 months after the flower falls.

The above-mentioned oral preparation in which the fresh leaves of the daylily are obtained from the daylily harvested 3 to 4 months after the flower falls.

The above-mentioned oral preparation, wherein the fermentation is brought about by a fermentation process using yeast.

The above-mentioned oral preparation in which the yeast is Saccharomyces cerevisiae or a subspecies thereof.

The above-mentioned oral preparation, which has been treated to inactivate the enzyme contained in the fresh leaves of the daylily, before or after being subjected to the fermentation treatment.

The above-mentioned oral preparation in which the treatment for inactivating the enzyme contained in the fresh leaves of the daylily is heat treatment or freezing treatment.

The above-mentioned oral preparation in which the temperature condition of the heat treatment is 60 to 100 ° C.

The oral preparation described above, wherein the time condition of the heat treatment is 10 seconds to 10 minutes.

The above-mentioned oral preparation in which the temperature condition of the freezing treatment is -4 ° C or less.

The above-mentioned oral preparation in which the temperature condition of the freezing treatment is -20 ° C or less.

The above-mentioned oral preparation in which the daily intake of the fermented leaf extract of Akinowasuregusa is 100 mg / day to 4000 mg / day in terms of dry solid content.

The above-mentioned oral preparation having an oxypinnatanine content of 2 mg to 20000 mg in 100 g of the fermented fresh leaf extract of the daylily daylily.

Use of fermented fresh leaf extract of Akinowasuregusa for the production of sleep quality improving agents.

The above-mentioned use in which the quality of sleep is reduction of drowsiness when waking up, recovery from fatigue when waking up, reduction of sleep onset time, increase of non-REM sleep, or reduction of awakening halfway.

The use described above, wherein the quality of sleep is a decrease in sleep onset time, an increase in non-REM sleep, and / or a decrease in awakening.

The above-mentioned use in which the water content of the fresh leaves of the daylily Akinowasu is 10% or more.

The above-mentioned use, in which the fresh leaves of the daylily are obtained from the daylily within 4 months after the flower falls.

The above-mentioned use, in which the fresh leaves of the daylily are obtained from the daylily harvested 1 to 4 months after the fall of flowers.

The above-mentioned use, in which the fresh leaves of the daylily are obtained from the daylily harvested 3 to 4 months after the flower fall.

The above-mentioned use, in which the fermentation is brought about by a fermentation process using yeast.

The above-mentioned use in which the yeast is Saccharomyces cerevisiae or a subspecies thereof.

The above-mentioned use, which is a treatment for inactivating the enzyme contained in the fresh leaves of the daylily, before or after being subjected to the fermentation treatment.

The above-mentioned use in which the treatment for inactivating the enzyme contained in the fresh leaves of the daylily is heat treatment or freezing treatment.

The above-mentioned use in which the temperature condition of the heat treatment is 60 to 100 ° C.

The above-mentioned use in which the time condition of the heat treatment is 10 seconds to 10 minutes.

The above use where the temperature condition of the freezing treatment is -4 ° C or less.

The above-mentioned use in which the temperature condition of the freezing treatment is -20 ° C or less.

The above-mentioned use in which the daily intake of the fermented leaf extract of the daylily Akinowas is 100 mg / day to 4000 mg / day in terms of dry solid content.

The above-mentioned use in which the content of oxypinnatanine in 100 g of the fermented fresh leaf extract of the daylily daylily is 2 mg to 20000 mg.

Fermented fresh leaf extract of daylily, which is used to improve sleep quality.

The above-mentioned extract whose sleep quality is reduction of drowsiness when waking up, recovery from fatigue when waking up, reduction of sleep onset time, increase of non-REM sleep, or reduction of awakening halfway.

The above extract, wherein the quality of sleep is a decrease in sleep onset time, an increase in non-REM sleep, and / or a decrease in awakening.

The above extract having a water content of 10% or more in the fresh leaves of the daylily.

The above extract, in which the fresh leaves of the daylily are obtained from the daylily within 4 months after the flower falls.

The above extract, in which the fresh leaves of the daylily are obtained from the daylily harvested 1 to 4 months after the flower falls.

The above extract, in which the fresh leaves of the daylily are obtained from the daylily harvested 3 to 4 months after the fall of flowers.

The above-mentioned extract obtained by the fermentation treatment using yeast.

The above-mentioned extract in which the yeast is Saccharomyces cerevisiae or a subspecies thereof.

The above extract, which has been treated to inactivate the enzyme contained in the fresh leaves of the daylily, before or after being subjected to the fermentation treatment.

The above extract, in which the treatment for inactivating the enzyme contained in the fresh leaves of the daylily is heat treatment or freezing treatment.

The above extract, the temperature condition of the heat treatment is 60 to 100 ° C.

The above extract, the time condition of the heat treatment is 10 seconds to 10 minutes.

The above extract, the temperature condition of the freezing treatment is -4 ° C or less.

The above extract, the temperature condition of the freezing treatment is -20 ° C or less.

The above-mentioned extract in which the daily intake of the fermented fresh leaf extract of the daylily is 100 mg / day to 4000 mg / day in terms of dry solid content.

The above-mentioned extract in which the content of oxypinnatanine in 100 g of the fermented fresh leaf extract of the daylily daylily is 2 mg to 20000 mg.

A method for improving sleep quality, including ingesting a fermented fresh leaf extract of Akinowasuregusa as an active ingredient.

The above method, wherein the quality of sleep is reduction of drowsiness when waking up, recovery from fatigue when waking up, reduction of sleep onset time, increase of non-REM sleep, or reduction of awakening halfway.

The method described above, wherein the quality of sleep is a decrease in sleep onset time, an increase in non-REM sleep, and / or a decrease in awakening.

The above method in which the water content of the fresh leaves of the daylily Akinowasu is 10% or more.

The above method, in which the fresh leaves of the daylily are obtained from the daylily within 4 months after the flower falls.

The above method, in which the fresh leaves of the daylily are obtained from the daylily harvested 1 to 4 months after the flower falls.

The above method, wherein the fresh leaves of the daylily are obtained from the daylily harvested 3 to 4 months after the flower fall.

The above method, wherein the fermentation is brought about by a fermentation process using yeast.

The above method in which the yeast is Saccharomyces cerevisiae or a subspecies thereof.

The above method, which is a treatment for inactivating the enzyme contained in the fresh leaves of the daylily, before or after being subjected to the fermentation treatment.

The above method, wherein the treatment for inactivating the enzyme contained in the fresh leaves of the daylily is heat treatment or freezing treatment.

The above method, wherein the temperature condition of the heat treatment is 60 to 100 ° C.

The above method, wherein the time condition of the heat treatment is 10 seconds to 10 minutes.

The above method in which the temperature condition of the freezing treatment is -4 ° C or less.

The above method in which the temperature condition of the freezing treatment is -20 ° C or less.

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

The above method, wherein the content of oxypinnatanine in 100 g of the fermented fresh leaf extract of the daylily is 2 mg to 20000 mg.

Next, the present invention will be specifically described with reference to Examples and Test Examples, but the present invention is not limited to the following Examples and Test Examples.

[Preparation of fermented fresh leaf extract of daylily]
The fresh leaves of Akinowasuregusa used were from Okinawa prefecture. Specifically, fresh leaves about 2 months after the fall of flowers were obtained from the daylilies cultivated in Okinawa Prefecture.

1 kg of fresh leaves of Akinowasuregusa was cut to a length of about 6 cm, crushed, squeezed, and purified by centrifugation to obtain squeezed juice. Yeast (Saccharomyces cerevisiae) is added to the juice concentrated to a Brix value of about 25% at a concentration of 0.5 to 1%, and fermented at 35 ° C to 38 ° C for about 3 hours. Was done. The fermented product was heated to 90 ° C. to inactivate the enzyme and then cooled to obtain a fermented leaf extract of Akinowasuregusa. The fermented fresh leaf extract of daylily was frozen under the condition of -18 ° C or lower in order to suppress the decomposition of oxypinnatanin.

In another embodiment, 1 kg of fresh leaves of daylily was cut into a length of about 6 cm, crushed, and then ground to form a paste. Yeast (Saccharomyces cerevisiae) was added to this paste at a concentration of 0.5 to 1%, and fermentation treatment was carried out at 35 ° C. to 38 ° C. for about 3 hours. The fermented product was heated to 90 ° C. to inactivate the enzyme and then cooled to obtain a fermented leaf extract of Akinowasuregusa. The fermented fresh leaf extract of daylily was frozen under the condition of -18 ° C or lower in order to suppress the decomposition of oxypinnatanin. In still another embodiment, it is possible to process 1 kg of fresh leaves of the daylily daylily into a paste by performing a fermentation treatment without cutting.

The present inventors have found that the amount of oxypinnatanin contained in the fresh leaves of daylily is decomposed and reduced due to the presence of sugars, and the fresh leaves of daylily are fermented to significantly contain oxypinnatanin. I was able to obtain an extract to be used. We also found that the presence of various enzymes contained in the fresh leaves of the daylily, oxypinnatanine, decomposed and reduced the amount of oxypinnatanin, and adopted a method of inactivating various enzymes by heat treatment. By producing by these methods, it was confirmed that the content of oxypinnatanine in 100 g of the fermented fresh leaf extract of Akinowasuregusa was 2 mg to 2000 mg at a high concentration.

[Test Example 1. Evaluation by clinical trial]
To investigate the sleep-improving effect of fermented fresh leaf extract of daylily, a randomized placebo-controlled parallel group study was conducted in 32 healthy subjects with sleep problems. As a placebo food, dextrin was used instead of the fermented leaf extract of Akinowasuregusa.

The subjects were healthy men and women between the ages of 20 and 65 who were mainly working at desk work and were not suffering from illnesses that were aware of mild sleep problems such as tiredness, poor sleep, and light sleep. ..

(Test method)
The study was conducted in a randomized, double-blind, placebo-controlled, parallel-group study. As a screening test, 40 subjects were visited to the hospital, and PSQI sleep questionnaire, blood test, urinalysis, and interview were performed, and 32 subjects who met the selection criteria of this study and did not meet the exclusion criteria were selected. As a pre-ingestion test, the selected subjects filled out a one-week OSA sleep questionnaire at the time of waking up and wore a sleep electroencephalograph (ZA-1, Pro Assist) on the day before the working day to measure sleep electroencephalogram. After the completion of the pre-ingestion test, the assigned test food was ingested 4 tablets a day for 2 weeks. During the intake period, the OSA sleep questionnaire was filled out every day at the time of waking up, and a diary describing the physical condition was kept every day. In addition, a sleep electroencephalograph was attached on the day before the working day of the second week of ingestion to measure the sleep electroencephalogram as in the case of the pre-intake examination. After the end of the ingestion period, the patient was returned to the hospital to perform a blood test and a urine test, as well as a PSQI sleep questionnaire and a post-intake test by interview.

The test beverages were randomly assigned by the person in charge of allocation who was not involved in the implementation of this study by the substitution block allocation method using age, gender, and PSQI as stratification factors. 16 people were assigned to each of the test products or the placebo food group, and ingestion was started. The intake period was 2 weeks. A sleep quality survey was conducted using the OSA sleep questionnaire during the ingestion period, and sleep electroencephalograms were measured in the second week of ingestion. The study was completed in all subjects and no one dropped out. However, the sleep electroencephalograph data could not be obtained for one person in the placebo intake group, so it was regarded as deficient.

The OSA sleep questionnaire is a psychological scale that evaluates sleep introspection when waking up, and is a questionnaire that determines the quality of sleep used in sleep outpatient clinics. The sleep questionnaire is divided into five factor components.

Regarding the results of blood test and urine test, no abnormal change was observed in both groups before and after ingestion. In addition, no adverse events related to the intake of the test food were observed.

This test showed the results as mean ± standard deviation. The significance level of the test was 5% on both sides, comparison between groups was performed by Student's t-test, and intra-group comparison was performed by paired t-test. Statistical analysis is performed by SAS Ver. It was carried out using 9.4 (manufactured by SAS). The results are shown in FIGS. 1 and 2. “*” In FIGS. 1 and 2 indicates that there is a significant difference at p <0.05.

(Result of OSA sleep questionnaire)
As shown in FIGS. 1 and 2, in the analysis of the OSA sleep questionnaire, the oxypinnatanine intake group derived from Akinowasuregusa (denoted as Kwansou in each figure) is significantly the I factor "sleepiness at wake up" as compared with placebo. And there was a significant difference in "recovery from fatigue" of factor IV, and ingestion of daylily-derived oxypinnatanine improved sleep quality.

(Results by sleep electroencephalograph)
Sleep was evaluated by measuring the brain waves of the subjects during bedtime. Sleep quality is assessed by a delta power value that indicates the amount of low frequency brain waves (delta waves) that appear during non-REM sleep. Non-REM sleep amount is evaluated by the length of non-REM sleep time during sleep. Sleep rhythm is evaluated by the length of time in the first sleep cycle (sleep onset-non-REM sleep-REM sleep). The results are shown in FIG.

As shown in FIG. 3, in the analysis of the sleep electrosurgical meter, the Akinowasregusa intake group was compared with the placebo intake group in terms of the ratio of the awakening time, the latency of falling asleep 2 weeks after the intake and the amount of change from before the intake. It decreased significantly. In addition, the rate of non-REM sleep was significantly increased in the daylily intake group compared with placebo.

The change in the rate of REM sleep (%) from before ingestion showed a significant decrease in the Akinowas regusa ingestion group compared with placebo.

[Test Example 2. Seasonal variation evaluation of oxypinnatanin in each part of daylily]
In order to effectively utilize medicinal plants used for food such as daylily, it is considered optimal to collect and use them when a large amount of active ingredient is contained. For that purpose, it is necessary to examine the change of the active ingredient in the growth process of the plant with time, and the change of oxypinnatanin (denoted as OPT in each figure) in the plant of the daylily daylily was examined.

The flowers, leaves, and roots of the daylily were collected in a raw state, extracted with methanol at room temperature for a whole day and night without drying, and then concentrated under reduced pressure to prepare an extract.

10 mg of the obtained extract was dissolved in purified water to make exactly 5 ml. 2.5 ml of it was adsorbed on the Sep-PAC C18 cartridge and eluted with 2 ml of purified water. Purified water was added to this eluate, and the total amount was adjusted to exactly 5 ml as a test solution, and OPT was detected by a fluorescence detection method using HPLC. The quantitative value was converted into the amount of OPT per 1 g of Fresh of each sample. (N = 3)

(HPLC conditions)
Pump: Shimadzu LC-10AS
Volume: CAPCELL PAK C ₁₈ (UG120), 150 x 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 advanced 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 x 0.5 mm (ID), 55 ° C.
Pump: JASCO 880-PU
Reaction reagent:
0.3MH ₃ BO ₃ , 3.0 mg / l 2-Mercaptoethanol,
40 mg / l o-Phthalaldehyde hyde
pH 10.5 advanced by NaOH solve.
Flow rate: 0.5 ml / min

Figure 4 shows the results of evaluating the seasonal variation of oxypinnatanine associated with flowering. The results of evaluating the fluctuation of oxypinnatanin before the flowering period in the leaves and roots are shown in FIG.

As shown in FIG. 4, oxypinnatanine in raw flowers was high in the bud's childhood and tended to decrease as it bloomed.

In addition, as shown in FIG. 5, it was confirmed that oxypinnatanine in raw leaves and roots showed a constant value until just before flowering and increased after flowering (after flowering period). After the flowering period in FIG. 5, the oxypinnatanine content increased remarkably within 1 month after the flowering (after the flowering period).

Oxypinnatanin is relatively stable when dissolved in water and at pH 1.2 to pH 6.8, suggesting that oxypinnatanin may be accumulated at high density in the early stages of plant organs and consumed during the growth process. Was done. Therefore, although not limited, it was confirmed that it is preferable to collect the fresh leaves of the daylily daylily within one month after the flower fall and prepare it as an extract without drying. In this case as well, it is more preferable to carry out the fermentation treatment using yeast, for example, yeast, in order to prevent the oxypinnatanine contained in the fresh leaves of the daylily from being decomposed and reduced due to the presence of sugars.

[Test Example 3. 28-day repeated oral administration toxicity test of fermented fresh leaf extract of daylily in rats]
The extract prepared as described above was pulverized by a conventional method and repeatedly orally administered to 5 male and female rats at a dose of 1000 mg / kg, which is the maximum dose specified in the guideline, for 28 days, and the presence or absence of toxicity was examined.

As a result, in the hematological examination, the activated partial thromboplastin time and the prothrombin time in the male test substance group were significantly prolonged as compared with the control group. However, this change was not observed in females, and no bleeding tendency was observed in the subcutaneous findings at autopsy in the male test substance group, and blood biochemical and histopathological changes suggesting liver abnormalities were observed. However, since it is unlikely that it inhibits the synthesis of a wide range of blood coagulation factors across the intrinsic and extrinsic systems, the significant thromboplastin time and prothrombin time of the activated partial thromboplastin observed in the male test substance group are significant. Prolongation was not considered to be a direct toxic reaction due to administration of the test substance.

Myocardial degeneration / fibrosis was observed in one male in the test substance-administered group. Although this change was a change in the test substance-administered group, it was a very slight change and no similar change was observed in other animals in the same group. Since there was no increase in AST) value, degeneration / fibrosis of the myocardium of this animal was considered to be an accidental finding.

As a result of organ weight measurement, significantly low values of absolute heart weight and relative weight were observed, but the degree of weight loss was negligible. In the histopathological examination of the heart, the above-mentioned slight myocardial degeneration / fibrosis was observed in one case, but the heart weight of this animal was rather higher than that of other animals in the same group. It was clear that there was no association between myocardial degeneration / fibrosis and cardiac weight loss. Since no abnormal histological findings other than these findings were observed, it was considered that the weight loss of the heart was not an organic change. On the other hand, the absolute weight of the lung was also significantly reduced as compared with the control group, but no abnormal histopathological findings were observed in the lung, and the weight loss was not considered to be a toxic change.

No other abnormalities were observed in the observation of the general condition, and no changes considered to be due to the administration of the test substance were observed in the body weight transition, food intake, ophthalmologic examination, urinalysis, and blood biochemical examination.

From the above, it was judged that the above extract powder was not toxic to rats when it was repeatedly administered at a dose of 1000 mg / kg for 28 days.

Claims

An oral preparation for improving sleep quality that contains fermented fresh leaf extract of daylily as an active ingredient.
The oral preparation according to claim 1, wherein the quality of sleep is reduction of drowsiness when waking up, recovery from fatigue when waking up, reduction of sleep onset time, increase of non-REM sleep, or reduction of awakening during sleep.
The oral preparation according to claim 1 or 2, wherein the water content of the fresh leaves of the daylily is 10% or more.
The oral preparation according to any one of claims 1 to 3, wherein the fresh leaves of the daylily daylily are obtained from the daylily daylily within 4 months after the flower falls.
The oral preparation according to any one of claims 1 to 4, wherein the daily intake of the fermented fresh leaf extract of the daylily is 100 mg / day to 4000 mg / day in terms of dry solid content.