JP6294870B2 - Sleep-promoting agent comprising yeast culture as active ingredient, oral composition for promoting sleep, and food composition for promoting sleep - Google Patents

Description

  The present invention relates to a yeast culture, an internal use composition and use thereof.

  Methylthioadenosine is known as a biological component. In Non-Patent Document 1, in sake brewing, S-adenosylmethionine and methylthioadenosine are contained in the yeast culture solution, and S-adenosylmethionine in fresh liquor is converted to methylthioadenosine when hot sake is stored by burning. It is described to do. Patent Document 1 describes that when a microorganism having S-adenosylmethionine-producing ability is cultured in a methionine-containing medium, methylthioadenosine is produced, and the maximum value is 0.0204 in terms of the amount of methylthioadenosine per dry cell. % (See comparative example of Patent Document 1).

  Non-Patent Document 2 describes that the addition of methylthioadenosine increases the cAMP concentration in S49 cells, and that the increase in cAMP concentration is involved in growth inhibition.

JP 58-146291 A

J. et al. Soc. Brew. Japan 70 (8) 585-587, 1975 Biochem Pharmacol. 33 (22): 3639-43. , 1984

  However, the methylthioadenosine content in non-patent literature 1 in sake and the methylthioadenosine content in the culture in patent literature 1 were both small. Further, Non-Patent Document 2 did not describe anything other than growth inhibition as an effect resulting from an increase in cAMP concentration.

  An object of the present invention is to provide a yeast culture and composition having a high methylthioadenosine content, and to provide a new use of the yeast culture and composition.

The present invention provides the following [1] to [12].
[1] A yeast culture containing methylthioadenosine or a salt thereof and having a methylthioadenosine content of 5.5% by mass or more per dry mass of the yeast culture.
[2] A yeast culture containing an S-adenosylmethionine degradation product.
[3] The yeast culture according to [2] above, wherein the S-adenosylmethionine degradation product contains methylthioadenosine.
[4] The yeast culture according to [3] above, wherein the content of methylthioadenosine per dry mass of the yeast culture is 5.5% by mass or more.
[5] The yeast according to any one of [1] to [4] above, wherein the yeast culture containing S-adenosylmethionine or a salt thereof is heated to decompose S-adenosylmethionine or a salt thereof. A method for producing a culture.
[6] Including yeast and methylthioadenosine or a salt thereof, and A / (A + B) is 1 / 10,000 or more and 1/2 or less [wherein A is the content of methylthioadenosine in the composition and B is the composition Each yeast content in the product is represented. ], An internal use composition.
[7] The internal use composition according to the above [6], wherein A / (A + B) is from 1/750 to 1/5.
[8] The internal use composition according to the above [6], wherein A / (A + B) is 1/100 or more and 1/5 or less.
[9] The internal use composition according to the above [6], wherein A / (A + B) is 5.5 / 100 or more and 1/5 or less.
[10] The yeast culture according to any one of [1] to [4] above and / or the internal use composition according to any one of [6] to [8] above as an active ingredient, Sleep promoter.
[11] The sleep promoter according to [10] above, wherein the amount of methylthioadenosine intake per adult is 0.01 to 1000 mg / day.
[12] A food and drink composition containing the sleep promoter according to [10] or [11].

Moreover, this invention can provide the following embodiment in other aspects.
(1) A sleep promoting method comprising administering the yeast culture according to any one of [1] to [5] or the internal use composition according to any one of [7] to [10].
(2) The yeast culture according to any one of the above [1] to [5] or the oral composition according to any one of the above [7] to [10], which is orally administered. )the method of.
(3) The sleep promoter of the yeast culture as described in any one of said [1]-[5] or the internal use composition as described in any one of said [7]-[10] is manufactured. Use for.

  According to the present invention, yeast cultures and compositions having a high methylthioadenosine content are provided. The yeast culture and composition of the present invention can promote sleep and induce good natural sleep.

FIG. 1 is a sleep progress diagram showing a general pattern of REM sleep and non-REM sleep. FIG. 2 is a graph showing the results of Experimental Example 1.

[Explanation of common terms]
In the present invention, the yeast is not particularly limited as long as it is a fungus that passes most of the life cycle as a single cell and buds or produces a partition wall to divide cells. Examples of the yeast include Saccharomyces genus yeast and Shizosaccharomyces genus yeast. Saccharomyces cerevisiae (budding yeast) and preferable.

  In the present invention, the yeast culture means a mixture obtained by culturing yeast in a medium. The mixture usually contains yeast cells, cell components (cell fragments), products produced by metabolism of yeast, and the like. The yeast may be either live cells or dead cells.

  The yeast may be cultured in a conventional manner according to the culture conditions suitable for the species to which the yeast strain used belongs. There is no restriction | limiting in particular in the culture medium used for culture | cultivation, The component utilized for the culture medium utilized for culture | cultivation of normal microorganisms, such as a carbon source, a nitrogen source, an inorganic salt, may be contained. Examples of the carbon source include glucose, sucrose, acetic acid, ethanol, molasses, sulfite pulp waste liquid, and the like. Examples of the nitrogen source include nitrogen-containing inorganic salts such as urea, ammonia, ammonium sulfate, ammonium chloride, and ammonium phosphate, and nitrogen-containing organic substances such as corn steep liquor (CSL), casein, yeast extract, and peptone. Examples of inorganic salts include phosphates such as lime superphosphate and ammonium phosphate; potassium salts such as potassium chloride and potassium hydroxide; magnesium salts such as magnesium sulfate and magnesium hydrochloride; zinc; copper; manganese; Can be mentioned. The medium may further contain components such as vitamins, nucleic acid-related substances, and amino acids. The medium preferably contains at least ethanol and an amino acid (for example, methionine). When the medium is a solid medium, agar or the like is usually added.

  The medium may be a solid medium or a liquid medium. Examples of the culture mode in the case of a liquid medium include batch culture, fed-batch culture, continuous culture and the like, and any of these may be adopted.

  The culture temperature may be between 6 ° C. and 35 ° C., usually 20 to 35 ° C., preferably 25 to 32 ° C. The culture time may usually be 1 day or more and preferably 2 days or more and 10 days or less.

  In the present invention, methylthioadenosine is also referred to as 5'-deoxy-5'-methylthioadenosine. 5'-deoxy-5'-methylthioadenosine may be in either the anti-type or the syn-type. In the following description, methylthioadenosine or a salt thereof may be abbreviated as MTA.

  The salt of methylthioadenosine is not particularly limited as long as it is a pharmacologically acceptable salt, and may be selected according to the dosage form and the like. Suitable salts of methylthioadenosine include, for example, acid addition salts. The acid addition salt may be any of an inorganic acid addition salt and an organic acid addition salt. For example, hydrochloride, sulfate, nitrate, carbonate, phosphate, formate, oxalate, citrate , Ascorbate, methanesulfonate, 1,4-butanedisulfonate, 1,5-pentanesulfonate and p-toluenesulfonate.

  In the present invention, S-adenosylmethionine has a structure in which adenosine and methionine are linked via a methylsulfonyl bond. The optical isomers of methionine include L-form, D-form, and DL-form. In the present invention, methionine in the structure of S-adenosylmethionine may be any of the optical isomers.

  The salt of S-adenosylmethionine is not particularly limited as long as it is a pharmacologically acceptable salt, and can be selected according to the dosage form and the like. Suitable salts of S-adenosylmethionine include, for example, acid addition salts and halides, and specifically include hydrochloride, sulfate, p-toluenesulfonate (tosylate), sulfuric acid / p. -Toluenesulfonic acid salt, methanesulfonic acid, trifluoromethanesulfonic acid, 1,4-butanedisulfonic acid, 1,5-pentanesulfonic acid, phosphate, chloride, bromide and the like. Of these, hydrochloride or tosylate is preferred.

  Yeast should just contain at least any one of methylthio adenosine and its salt, and may contain both.

[First yeast culture of the present invention]
A preferred embodiment of the yeast culture of the present invention includes a yeast culture containing methylthioadenosine or a salt thereof.

  The content of methylthioadenosine or a salt thereof in the yeast culture can be shown as the content of methylthioadenosine per dry mass of the yeast culture. The content (content rate) of methylthioadenosine per dry mass means the percentage of the ratio of the net mass of methylthioadenosine contained in the yeast culture to the total amount of the yeast culture. If the yeast culture contains methylthioadenosine salt, the “net mass of methylthioadenosine” does not include the mass of the salt portion of methylthioadenosine salt. The methylthioadenosine content is preferably 5.5% by mass or more, more preferably 7% by mass or more, and further preferably 8% by mass or more. Thereby, the yeast culture of this invention can exhibit the physiological effect of methylthio adenosine or its salt efficiently. The upper limit of the methylthioadenosine content is usually 50% by mass or less, preferably 40% by mass or less, more preferably 30% or less, still more preferably 20% by mass or less, and still more preferably 15% by mass. It is as follows.

  Moreover, the high sleep induction effect is acquired by making the minimum of methylthio adenosine content per dry mass of a yeast culture into 5.5 mass% or more.

The methylthioadenosine content per dry mass in the yeast culture can be defined, for example, by the following formula (1).
Formula (1)
Methylthioadenosine content per dry mass of yeast culture = (a / b) × 100

  A in formula (1) is the content of methylthioadenosine (not including the mass of the salt portion of the methylthioadenosine salt) (g) in a certain amount collected from the yeast culture. Quantification as methylthioadenosine can be performed by liquid chromatography. b is the dry weight (g) after 24 hours of freeze-drying (trap cooling temperature −75 ° C.) of the same amount of yeast culture collected at the time of the above a measurement.

  In the yeast culture containing methylthioadenosine or a salt thereof, the yeast may contain methylthioadenosine or a salt thereof, or may contain methylthioadenosine or a salt thereof outside the yeast cells.

  The first yeast culture of the present invention is presumed to contain some component that improves the physiological activity of methylthioadenosine or a salt thereof (for example, intracellular cAMP concentration increasing activity, sleep promoting activity, etc.). Therefore, the 1st yeast culture of this invention can be used for various uses which exhibit said physiological activity.

[Second yeast culture of the present invention]
Another preferred embodiment of the yeast culture of the present invention includes a yeast culture containing a degradation product of S-adenosylmethionine or a salt thereof.

  The degradation product of S-adenosylmethionine or a salt thereof means a component produced by decomposition of S-adenosylmethionine and a component produced by decomposition of a salt of S-adenosylmethionine. Examples of the decomposition product of S-adenosylmethionine or a salt thereof include, for example, methylthioadenosine, S-adenosylhomocysteine, S-adenosylmethioniamine, S-aminobutyrolactone, adenosine, methionine, adenine, inosine, homoserine, homo Examples include cysteine and cystathionine. The degradation product of S-adenosylmethionine may be a single component or a combination of two or more components. The degradation product of S-adenosylmethionine is preferably methylthioadenosine, or preferably contains at least methylthioadenosine, more preferably methylthioadenosine or a salt thereof.

  The decomposition product of S-adenosylmethionine or a salt thereof may be a decomposition product of S-adenosylmethionine, a decomposition product of a salt of S-adenosylmethionine, or a decomposition product of S-adenosylmethionine and S- A combination of decomposition products of adenosylmethionine salt may be used.

  The degradation conditions in the degradation product of S-adenosylmethionine or a salt thereof are not particularly limited. Examples of the decomposition include decomposition by heating, decomposition by enzyme, acid decomposition, alkali decomposition, decomposition by chemical reaction, etc. Among these, decomposition by heating is preferable.

  The content of the degradation product of S-adenosylmethionine or a salt thereof in the yeast culture can be shown as the content of methylthioadenosine contained in the degradation product per dry mass of the yeast culture. The content (content rate) of methylthioadenosine per dry mass means the percentage of the ratio of the net mass of methylthioadenosine contained in the yeast culture to the total amount of the yeast culture. If the yeast culture contains methylthioadenosine salt, the “net mass of methylthioadenosine” does not include the mass of the salt portion of methylthioadenosine salt. The methylthioadenosine content per dry mass in the yeast culture of the present invention is preferably 5.5% by mass or more, more preferably 7% by mass or more, and further preferably 8% by mass or more. Thereby, the yeast culture of this invention can exhibit the physiological effect of methylthio adenosine or its salt efficiently. The upper limit of the methylthioadenosine content is usually 50% by mass or less, preferably 40% by mass or less, more preferably 30% or less, still more preferably 20% by mass or less, and still more preferably 15% by mass. It is as follows. The measurement of the methylthioadenosine content per dry mass in the yeast culture is as described in the section of the first yeast culture of the present invention.

  The yeast culture containing the degradation product of S-adenosylmethionine or a salt thereof may contain a degradation product of S-adenosylmethionine or a salt thereof. A degradation product of silmethionine or a salt thereof may be contained.

  The second yeast culture of the present invention is a physiological activity of a degradation product of S-adenosylmethionine or a salt thereof (for example, an intracellular cAMP concentration increasing activity, a sleep promoting activity, etc., which is a physiological activity of methylthioadenosine therein) ) Is presumed to contain some component that improves. Therefore, the 2nd yeast culture of this invention can be used for various uses which exhibit said physiological activity.

[Method for Producing Yeast Culture of the Present Invention]
Although the manufacturing method of the yeast culture of this invention is not specifically limited, It can manufacture simply by heating the yeast culture containing S-adenosyl methionine or its salt, The said method is preferable. Both the first yeast culture and the second yeast culture of the present invention are preferably produced by the method described above.

  The yeast containing S-adenosylmethionine or a salt thereof may be a yeast that originally contains S-adenosylmethionine or a salt thereof, or a yeast that does not originally contain S-adenosylmethionine or a salt thereof. Yeast obtained by producing S-adenosylmethionine may also be used. As an example of a method for producing S-adenosylmethionine or a salt thereof in yeast, a method of culturing yeast and producing S-adenosylmethionine or a salt thereof in the yeast (intracellular) can be mentioned. In the case of such a method, the obtained culture solution may be used as it is as a yeast culture containing S-adenosylmethionine or a salt thereof, or the yeast recovered after the culture is treated with S-adenosylmethionine or a salt thereof. You may use as yeast to contain. Furthermore, a dried yeast culture obtained by drying the culture medium after culturing as it is or after collecting the bacteria may be used. In the present invention, it is preferable to use a yeast culture containing S-adenosylmethionine or a salt thereof in a dry state.

  In addition, components such as a stabilizer and an excipient may be added to the yeast culture weighed by the formula (1), and then the following heating operation may be performed.

  Conditions for heating the yeast culture containing S-adenosylmethionine or a salt thereof are not particularly limited. The temperature is usually 40 ° C. or higher, preferably 50 ° C. or higher, more preferably 60 ° C. or higher, and still more preferably 80 ° C. or higher. The upper limit is usually 200 ° C. or lower, preferably 180 ° C. or lower, more preferably 160 ° C. or lower, and still more preferably 120 ° C. or lower. The temperature is usually 40 ° C. or higher and 200 ° C. or lower, preferably 50 ° C. or higher and 180 ° C. or lower, more preferably 60 ° C. or higher and 160 ° C. or lower, and still more preferably 80 ° C. or higher and 120 ° C. or lower. The treatment time may be adjusted depending on the degree of formation of methylthioadenosine or a salt thereof.

  There is no particular limitation on the heating method. As a heating method in the case of a liquid yeast culture, for example, a method in which a container in which the yeast culture is put is heated while stirring in a heating apparatus such as an electric heater or an electric heater, and a container in which the yeast culture is put in a hot water bath. , A method of heating in an oil bath or the like while stirring, a method of heating the yeast culture in a water jacketed container and heating it through hot water or steam in the water jacket, and a container containing the yeast culture with a heat exchange type plate heater Heating method, sealing the container containing the yeast culture, heating the container with hot water, heating with electromagnetic waves (microwave oven), heating with spray drying, heating with autoclave, fluidized bed The method of heating with dryers, such as a dryer and a shelf dryer, etc. are mentioned. Examples of the heating method in the case of a solid yeast culture such as powder include a method of heating in an autoclave, a method of heating a sealed container with hot water, a fluidized bed dryer, a rotary aeration dryer, and a shelf dryer. And a method of heating with a dryer such as a method of heating with a roaster.

  A yeast culture with a high content of methylthioadenosine or a salt thereof can be obtained by drying the yeast culture following the heating or standing treatment. The method for drying the culture is not particularly limited, and any of the methods commonly used for preparing dry yeast cells may be used. For example, freeze-drying method, spray-drying method And drum drying method. Furthermore, by processing the obtained dry yeast culture into a powder form, a yeast culture having a high content of methylthioadenosine or a salt thereof excellent in handleability can be obtained.

[Internal use composition of the present invention]
The internal use composition of the present invention contains yeast and methylthioadenosine or a salt thereof.

  Methylthioadenosine or a salt thereof can be obtained by a known method, and may be a commercially available product. Examples of the commercially available product of methylthioadenosine or a salt thereof include a commercially available product manufactured by Sigma-Aldrich and a commercially available product manufactured by Santa Cruz, and the former is preferred. In addition, part or all of methylthioadenosine may be contained in yeast.

  The yeast may be live bacteria, dead bacteria, or bacterial cell components. The yeast may be a commercial product.

  One type of yeast may be used alone, or two or more types having different varieties and forms may be used. Each of methylthioadenosine and its salt may be one kind alone, or may be a combination of two or more kinds such as chemical structure, kind of salt and form. The internal use composition should just contain methylthio adenosine and / or its salt, and may contain both.

In the internal use composition of the present invention, the mass ratio represented by the following formula (2) is preferably 1 / 10,000 or more and 1/2 or less, more preferably 1/750 or more and 1/5 or less, It is further preferably 1/100 or more and 1/5 or less, and further preferably 5.5 / 100 or more and 1/5 or less. Thereby, the outstanding sleep promotion effect can be acquired. When A / (A + B) is 5.5 / 100 or more and 50/100 or less, both the bitter taste of methylthioadenosine and the residual feeling in the mouth of yeast can be reduced:
Formula (2)
A / (A + B)

  In formula (2), A is the content of methylthioadenosine in the composition. By methylthioadenosine content in the composition is meant the net amount of methylthioadenosine contained in the composition. When the oral composition contains methylthioadenosine salt, the methylthioadenosine content does not include the mass of the salt portion of methylthioadenosine salt. When the yeast contains methylthioadenosine or a salt thereof, the net amount of methylthioadenosine contained in the yeast is also reflected in the methylthioadenosine content. In formula (2), B is the yeast content in the composition.

  In producing the internal composition of the present invention, yeast and methylthioadenosine or a salt thereof may be mixed, and the production conditions are not particularly limited. When mixing, a solvent (such as an aqueous methylcellulose solution) may be used.

  The presence form of yeast and methylthioadenosine or a salt thereof in the internal use composition of the present invention is not particularly limited. Usually, the aspect in which yeast contains part or all of methylthioadenosine or a salt thereof may be employed.

  The internal use composition of the present invention can improve the physiological activity of methylthioadenosine or a salt thereof (for example, intracellular cAMP concentration increasing activity, sleep promoting activity) by a combination of yeast and methylthioadenosine or a salt thereof. Guessed. Therefore, the internal use composition of this invention can be used for various uses which exhibit said physiological activity.

[Sleep Accelerator]
Since the yeast culture and the internal use composition of the present invention exhibit a natural sleep promoting effect, each or a combination of both is a sleep promoting agent (sleep promoting composition), particularly a non-REM sleep promoting agent, more preferably deep. It is useful as an active ingredient of a sleep promoting agent (deep sleep promoting composition).

  The sleep promoting effect means an effect of extending the ratio of the non-REM sleep time to the sleep time. Non-REM sleep can be determined by polysomnograph analysis (eg, electroencephalogram, electrooculogram, electromyogram). As an electroencephalogram, non-REM sleep can be determined by a decrease in alpha waves (for example, 8 to 3 Hz in humans) and an increase in delta waves (for example, 0.5 to 4.5 Hz in humans). The measured electroencephalogram can also be determined using automatic analysis software such as SleepSign (registered trademark). It is preferable that the ratio of the deep sleep time is high in the non-REM sleep time.

  “Promoting non-REM sleep” means that 1) the state of non-REM sleep is observed for a longer period of time compared to the state to be compared (for example, the state before ingestion of the internal use composition of the present invention), 2) It means that the effect of either deeper non-REM sleep or 3) the smooth transition to non-REM sleep is observed.

  “Deep sleep” means sleep in which the depth of sleep is a certain level or more in the case of humans, and the depth of layer III or higher when non-REM sleep is classified into four stages of I to IV layers (FIG. 1). It means the sleep state in During sleep of layer III and layer IV, the occupancy of the delta wave (0.5 to 4.5 Hz) with respect to the entire electroencephalogram (0.5 to 20 Hz) is more than a certain level. It is known that the power value is high. Therefore, deep sleep can also be defined as sleep with a high delta power value. The delta power value can be measured using, for example, a polysomnography inspection apparatus, and the “depth of sleep”, which is the sum of sleep depth and time, is obtained by multiplying the delta power value by time. Defined.

  “Promoting deep sleep” means that 1) deep sleep is observed for a longer period of time compared to the state to be compared (for example, the state before administration of a sleep enhancer), 2) depth of deep sleep It means that the effect of either deepening the depth or 3) the smooth transition to deep sleep is observed. That is, in an experimental system using a mouse or a guinea pig, a value obtained by multiplying the delta power value by time may be increased.

  “Natural sleep” refers to sleep that does not cause loss of direct reflex. “Direct reflection” is a reflection motion that restores the head to a normal position with respect to the direction of gravity, and is also referred to as bounce reflection.

  Loss of specular reflex in rodent animals such as mice and rats can be confirmed, for example, as a state in which a mouse or rat placed in the supine or dorsal position does not return to the prone position within 30 seconds. When the sleeping animal is placed in a supine or dorsal position, if it immediately wakes up from sleep or quickly returns to the prone position, the sleep does not lose its direct reflex It is sleep. In humans, there is no equivalent of “direct reflection” as defined by animals. However, in humans, the state of “not losing the right-angled reflection” corresponds to sleep that can be easily awakened, for example, when the shoulder is struck, and can be defined as natural sleep. it can.

  In contrast to natural sleep, anesthesia causes the loss of direct reflexes. Loss of direct reflex in mice or rats is known as an evaluation index of anesthetic effect. The anesthetic effect of an anesthetic such as pentobarbital or other drugs can be evaluated by the increase in the number of animals that have lost the right reflex. In addition, the frequency distribution analysis by Fourier transform analysis of the brain wave during anesthesia or sleeping with a sleeping drug as a conventional medicine, without anesthesia or using a conventional sleeping drug, hypnosis with anesthesia or a conventional sleeping drug, and these In the case of sleep without using an anesthesia, both delta wave (0.5-4.5 Hz) electroencephalograms increase, but in hypnosis with anesthesia or conventional hypnotics, sleep without using anesthesia or conventional hypnotics at 15-20 Hz. An increase in brain waves in the frequency band that is sometimes not seen is observed. Although it has not been clarified at present what causes the electroencephalogram in the frequency band that cannot be seen during sleep without anesthesia or conventional hypnotics, there is a relationship with anesthesia or poor awakening due to conventional hypnotics. Inferred.

  The content of the yeast culture in the sleep promoter of the present invention is not particularly limited, and can be appropriately adjusted according to various conditions such as an effective amount that can exert the effect. There is no restriction | limiting in particular in content of the internal use composition in the sleep promoting agent of this invention, According to various conditions, such as an effective amount which can exhibit an effect, it can adjust suitably.

  The sleep promoting agent of the present invention may be mixed or used in combination with components (including sleeping pills) that can promote sleep other than the yeast culture and internal use composition of the present invention. Here, as a sleeping pill, what is called a sleep improving agent, a sleep inducer, etc. may be contained, for example. However, the mixed form with the kind or quantity of an anesthetic that causes the disappearance of the direct reflex is excluded.

  The sleep-promoting agent of the present invention comprises a preparation consisting essentially only of the yeast culture and / or internal composition of the present invention, or the yeast culture and / or internal composition of the present invention, the yeast culture and the Other ingredients other than the internal use composition can be made into a preparation (composition) containing one or more kinds.

  The said other component contained in the sleep promoting agent of this invention is not specifically limited unless the effect of this invention is impaired essentially. Examples of the other components include excipients, disintegrants, binders, lubricants, coating agents, colorants, color formers, corrigents, flavoring agents, antioxidants, preservatives, flavoring agents, and acidity. Pharmacologically acceptable additives such as agents, sweeteners, fortifiers, vitamins, swelling agents, thickeners, and surfactants. Among these, the effect of promoting sleep, various properties necessary for the preparation (for example, preparation stability) are not impaired, and the final product (for example, food and drink, pharmaceutical, quasi-drug, nutritional supplement (supplement) ) Etc.) may be selected from one or more additives depending on the dosage form. The other component contained in the sleep promoting agent of the present invention may be one type or a combination of two or more types.

  The dosage form of the sleep promoting agent of the present invention is not particularly limited as long as the yeast culture and / or internal use composition of the present invention can be used as an active ingredient. Examples of dosage forms for oral administration include liquid (liquid), syrup (syrup), solid (tablet), capsule (capsule), powder (granule, fine granule), soft capsule ( Soft capsules), semi-liquid, cream, and paste. Examples of dosage forms for parenteral administration include liquids (injections, nasal drops), mists (sprays, inhalants) and the like. These formulations can be prepared by mixing the yeast culture of the present invention with a pharmacologically acceptable medium.

  The dosage of the sleep promoting agent of the present invention is not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately adjusted depending on factors such as the age and condition of the administration target organism. The preferable dose for obtaining the desired effect is the dose (mg / day) of methylthioadenosine or a salt per day for an adult, and the lower limit thereof is preferably 0.01 mg or more, more preferably 0.1 mg or more. More preferably, it is 1 mg or more, and still more preferably 10 mg or more. The upper limit becomes like this. Preferably it is 1000 mg or less, More preferably, it is 500 mg or less, More preferably, it is 100 mg or less.

  The administration mode of the sleep promoting agent of the present invention is not particularly limited and may be either oral administration or parenteral administration, but oral administration is preferred because of less invasiveness. Examples of oral administration include oral administration and sublingual administration. Examples of parenteral administration include intravenous administration, intramuscular administration, subcutaneous administration, transdermal administration, nasal administration, and pulmonary administration.

  Although the time of administration of the sleep promoting agent of the present invention is not particularly limited, it is usually administered before bedtime, preferably administered from 3 hours before bedtime to bedtime, and administered between 2 hours before bedtime and bedtime. More preferably, it is more preferably administered from 1 hour before bedtime to bedtime, and even more preferably administered 1 hour before bedtime.

  The ingestion subject of the sleep promoting agent of the present invention is not particularly limited. For example, people who are ingested may feel sleepy, sleepy when waking up, poor sleep, lack of deep sleep (cannot sleep deeply), bad dreams, and fatigue after sleep. Examples include subjects who are dissatisfied and subjects who feel fatigue and want to improve their subjective sleep feeling. Moreover, the subject who does not have a special problem can be ingested on a daily basis for the purpose of maintaining good sleep, preventing sleep disorders, and promoting natural sleep more.

  The sleep promoting agent of the present invention may be used as an additive for food and drink, a pharmaceutical additive, or an quasi-drug additive. Thereby, the effect which promotes natural sleep can be provided to food-drinks, a medicine, and a quasi-drug.

[Food and Drink of the Present Invention]
The food / beverage products containing the sleep promoting agent of the present invention can be used as a food / beverage product expected to have the sleep promoting effect as described above and the effects associated therewith. Examples of such foods and beverages include health foods, functional foods, dietary supplements (supplements), foods for specified health use, medical foods, foods for the sick, foods for infants, foods for nursing care, foods for the elderly Etc.

  The sleep promoting agent of the present invention can increase the non-REM sleep time during sleep as compared to the case where the sleep promoting agent of the present invention is not administered, as shown in the following examples. Therefore, application to the prevention and treatment of diseases associated with reduced sleep quality by shortening non-REM sleep time, decreasing sleep depth, or both is also expected.

  Since the sleep promoting agent of the present invention can promote natural sleep, it can also be used as a natural sleep inducing agent. “Inducing natural sleep” refers to inducing sleep without loss of direct reflex. Since the natural sleep inducer of the present invention can promote natural sleep, it can contribute to maintaining a good sleep pattern or restoring a disturbed sleep pattern to a good sleep pattern. When using the sleep promoting agent of the present invention as a natural sleep inducing agent, the embodiment of the dosage may be applied to the embodiment of the sleep promoting agent of the present invention.

[Food and Drink of the Present Invention]
There is no restriction | limiting in particular in the form of the food / beverage products of this invention, For example, a drink (a soft drink, a carbonated drink, a nutritive drink, a powdered drink, a fruit drink, a milk drink, a jelly drink etc.), confectionery (cookie, cake, gum, candy) , Tablets, gummi, buns, sheepskin, pudding, jelly, ice cream, sorbet, etc.), processed fishery products (kamaboko, chikuwa, hanpen, etc.), livestock products (hamburger, ham, sausage, winner, cheese, butter, yogurt, Fresh cream, margarine, fermented milk, etc.) soup (powder soup, liquid soup, etc.), staple foods (rice, noodles (dried noodles, raw noodles), bread, cereal, etc.), seasonings (mayonnaise, shortening, dressing, Source, sauce, soy sauce, etc.). Furthermore, the food and drink of the present invention are health food, functional food, health supplement (supplement), nutrition supplement, specific health food, medical food, sick food, infant food, nursing food, elderly It may be a food or drink such as food for consumers. Of these, health supplements are preferred, and tablet-like health supplements are more preferred.

  EXAMPLES Hereinafter, although an Example demonstrates this invention concretely, this invention is not limited to the following Example.

Examples 1 to 13 and Comparative Examples 1 to 3
<Sample preparation>
In Examples 1-2, 8-13, and Comparative Examples 2-3, yeast (Nisshin Foods Co., Ltd., trade name: Nisshin Super Camellia Dry Yeast), methylthioadenosine or a salt thereof (hereinafter referred to as MTA). Sigma-Aldrich, trade name: 5′-Deoxy-5 ′-(methylthio) adenosine) was mixed so that the mass ratio of MTA in the entire mixture was as shown in Table 1. The resulting composition was used as a sample.

  In Examples 3 to 7, yeast cultures each containing MTA (hereinafter referred to as MTA-containing yeast culture) were used. The preparation method of the MTA-containing yeast culture was as follows. A yeast culture powder containing S-adenosylmethionine or a salt thereof (manufacturer name: Iwata Chemical Co., Ltd., trade name: Amy) is heated at 120 ° C. for 20 minutes using a dryer, and MTA-containing yeast culture is obtained. Got. The mass ratio of MTA in each example is as shown in Table 1.

Footnote * in Table 1: “Mass ratio” is the ratio of methylthioadenosine content to the total of methylthioadenosine content and yeast content when the sample is a composition. When the sample is an MTA-containing yeast culture, it is the mass ratio of MTA in the yeast culture.

  Note that the methylthioadenosine content of the yeast culture of the sample is defined by the above formula (1). For example, the sample of Example 1 was measured by a known method capable of separating S-adenosylmethionine and MTA by high performance liquid chromatography. The methylthioadenosine content when the sample is a composition is defined by equation (2) above.

<Experimental example 1>
Each sample of Examples 1-13 and Comparative Examples 1-5 was allowed to act on HEK cells (Perkin Elmer, cAMPZen (cultured cells for cAMP assay)) for 30 minutes. Thereafter, in order to measure intracellular cAMP concentration by FRET (fluorescence resonance energy transfer), Eu-cAMP tracer and ULlight-anti-cAMP (both manufactured by Perkin Elmer, reagents in LANCE Ultra cAMP Kit) were added. Incubated for hours. Thereafter, the signal value was measured under the following conditions. The intracellular cAMP concentration increasing activity was calculated by the following formula (3).

<Measurement conditions>
Excitation wavelength: 320 nm
Emission wavelength: 665 nm
Integration time: 100μs
Lag time: 50 μs

<Calculation formula>
Formula (3):
Intracellular cAMP concentration increasing activity% =
{(Fluorescence signal value when no sample is added−Sample fluorescence signal value) /
(Fluorescence signal value when no sample is added-fluorescence signal value when MTA is administered alone)}
× 100

  The result of the intracellular cAMP concentration increasing activity of each Example is shown in FIG. The following is apparent from FIG.

  In Examples 1, 2, and 8 to 13, the activity was higher than that of Comparative Examples 1 to 5. This indicates that the composition of the present invention has an effect of increasing intracellular cAMP concentration.

  Examples 3-7 were more active than Comparative Examples 1-5. This indicates that the yeast culture of the present invention has an effect of increasing the intracellular cAMP concentration.

  Examples 7 and 8 were more active than Example 8 although Example 7 and 8 had the same MTA mass ratio. This indicates that when MTA is contained in yeast, there is an effect of significantly increasing the intracellular cAMP concentration more than when yeast is added to MTA.

  It is considered that when the intracellular cAMP concentration in the sleep center increases, sleep is promoted. From this, it is estimated that the composition and yeast culture of the present invention exhibit a sleep promoting effect.

Example 14 and Comparative Examples 6-8
<Evaluation sample and dose>
As a sample of Comparative Example 6 (control solution), a 0.5% by mass methylcellulose aqueous solution was used. The dose was 10 mL / kg body weight.

  The sample of Comparative Example 7 is MTA (trade name: 5'-Deoxy-5 '-(methylthio) adenosine) manufactured by Sigma-Aldrich, 0.5% by mass methylcellulose (trade name, Methyl cellulose, manufactured by Sigma-Aldrich). Prepared by suspending in an aqueous solution. The dose was 15 mg / kg body weight as the amount of MTA (10 mL / kg body weight as a suspension).

  The sample of Example 14 was prepared as follows. Commercially available S-adenosylmethionine-containing yeast culture powder (manufacturer name: Iwata Chemical Co., Ltd., trade name: Amy) was heat-treated at 120 ° C. for 20 minutes. The methylthioadenosine content in the obtained yeast culture was 275 mg, the dry weight of the yeast culture (including MTA) was 5000 mg, and the dry weight of the yeast cultures other than MTA was 4725 mg. That is, the mass ratio of the methylthioadenosine content to the yeast culture (expressed by the above formula (1)) is 5.5% by mass, and the ratio expressed by the above formula (2) is 275 / (275 + 4725). It was 5.5 / 100 (about 6/100). The resulting product was suspended in a 0.5 mass% methylcellulose aqueous solution, and the suspension was used as a sample. The dose was 275 mg / kg body weight as yeast culture (10 mL / kg body weight as suspension, 15 mg / kg body weight as MTA amount).

  The sample of Comparative Example 8 is a sample containing yeast containing S-adenosylmethionine, and was prepared as follows. The same commercially available S-adenosylmethionine-containing yeast culture powder as used in Example 14 was suspended in a 0.5 mass% aqueous methylcellulose solution, and the suspension was used as a sample. The dosage was 275 mg / kg body weight as S-adenosylmethionine-containing yeast (10 mL / kg body weight as suspension, 0 mg / kg body weight as MTA amount).

<Experimental example 2>
Electroencephalogram and myoelectric electrodes were attached to 8-week-old male C57BL / 6 mice purchased from Japan SLC. After the electrodes were mounted, they were recovered for 10 days in a recovery chamber. Then, it moved to the recording chamber and the cable was connected. The brain wave analysis software SleepSign (registered trademark) Ver 3.0 (Kissei Comtech Co., Ltd.) was used for acclimatization for 3 days after confirming whether or not the brain wave could be discriminated. Thereafter, the electroencephalogram was measured for 24 hours to confirm whether or not the sleep / wake rhythm was maintained, and the administration groups of each Example and Comparative Example were performed. Each administration group consisted of 8 animals (n = 8).

  Immediately before the start of the dark period, each sample was orally administered to each administration group, and the electroencephalogram was recorded for 24 hours. In addition, since all the mice gavaged 275 mg / kg body weight as yeast cultures on the day different from the day on which the electroencephalogram measurement was performed, when touched to make the supine position, they showed arousal, It was confirmed that these sleeps were in a state that “the direct reflex was not lost”.

  After recording, the electroencephalogram was automatically analyzed by SleepSign (registered trademark) Ver 3.0, and the evaluator confirmed the result of the automatic analysis and classified it into sleep stages of wakefulness, REM sleep and non-REM sleep. The total integrated amount (unit: sec) of the non-REM sleep time from the EEG measurement time to 4 hours later is calculated, the percentage of the non-REM sleep amount at the time of non-administration (baseline) of the same individual is obtained, and the average value is shown in Table 2 It is shown in the column of “sleep amount”.

  As is clear from the results in Table 2, in Example 14, the ratio of the non-REM sleep amount at the time of administration to the non-REM sleep amount at the time of non-administration was very high as compared with Comparative Examples 6-8. This has shown that the yeast culture of this invention has the outstanding sleep improvement effect.

Examples 15-19
<Preparation of evaluation sample>
Each sample was a yeast culture containing MTA in the amounts shown in Table 3, and was prepared as follows. Yeast (Nisshin Foods Co., Ltd., trade name: Nisshin Super Camellia Dry Yeast) and methylthioadenosine or a salt thereof (hereinafter referred to as MTA. Sigma-Aldrich, trade name: 5′-Deoxy-5′- (Methylthio) adenosine) was mixed so that the mass ratio of MTA in the entire mixture was as shown in Table 3. The resulting composition was used as a sample.

<Experimental example 3>
After 10 subjects each orally ingested 50 mg of the sample, the degree of bitterness and the residual feeling in the mouth were evaluated according to the following criteria, and the average score of each panel was determined. In the evaluation of bitterness, the bitterness of MTA itself was used as a reference, and this was evaluated as one point. In the evaluation of the residual feeling in the mouth, the residual feeling with only yeast was used as a reference, and this was evaluated as one point.

Evaluation criteria for bitterness (score criteria)
5 points: feel no bitterness 4 points: feel bitterness 3 points: feel bitterness 2 points: feel bitterness strongly 1 point: feel bitterness unbearable

〔Evaluation criteria〕
◎: Average value of 3.5 points or more ○: Average value of 2.5 points or more and less than 3.5 points △: Average value of 1.5 points or more and less than 2.5 points ×: Average value of 1.0 points or more to 1.5 Less than

・ Evaluation criteria for feeling in the mouth 5 points: Feeling no remaining in the mouth 4 points: Feeling somewhat remaining in the mouth 3 points: Feeling remaining in the mouth 2 points: Feeling strongly in the mouth 1 point: In the mouth I feel unbearable to survive

  The average value of the 10 scores of the residual feeling in the mouth calculated according to the above evaluation criteria was determined, and the residual feeling in the mouth was determined based on the numerical value. The results are shown in Table 3. In Table 3, the mass ratio of MTA is the mass ratio of MTA to yeast culture.

  From the results of Table 3, in Examples 16 to 18, both the bitterness derived from MTA and the residual feeling in the mouth were reduced. This has shown that the yeast culture of this invention exhibits favorable flavor, when MTA content is 0.06-0.5 mass%.

Example 20
<Preparation of evaluation sample>
The MTA-containing yeast culture was prepared as follows. Commercially available S-adenosylmethionine-containing yeast culture powder (manufacturer name: Mitsubishi Gas Chemical Co., Ltd., trade name: SAMe-containing dry yeast) was heat-treated at 120 ° C. for 20 minutes. The methylthioadenosine content in the obtained yeast culture was 320 mg, the dry weight of the yeast culture (including MTA) was 5000 mg, and the dry weight of the yeast cultures other than MTA was 4680 mg. That is, the mass ratio of the methylthioadenosine content to the yeast culture (expressed by the above formula (1)) is 6.4% by mass, and the ratio expressed by the above formula (2) is 320 / (4680 + 320). It was 6.4 / 100.

  As a sample of Example 20, a tablet (methylthioadenosine content in 4 tablets: 21 mg) containing the above MTA-containing yeast culture was prepared. As a sample of Comparative Example 9, a placebo tablet not containing MTA-containing yeast was prepared. Details of each composition are as shown in Table 4.

<Experimental example 4> (Evaluation in human)
Six subjects were orally ingested 4 tablets of the sample of the above Example 20 one hour before going to bed, and a two-electrode portable electroencephalograph (“Electroencephalograph Sleep Scope” manufactured by Sleep Well Co., Ltd.) was used. The brain waves were measured while the subject was sleeping. Similarly, the same six subjects were ingested four tablets of the sample of Comparative Example 9 one hour before going to bed, and a two-electrode portable electroencephalograph (EEG) manufactured by Sleep Well Co., Ltd. A total sleep scope ") was worn, and the brain waves during the sleep of the subject were measured. For each subject, administration and electroencephalogram measurement were performed four times for each of the sample of Example 20 and the sample of Comparative Example 9.

  Moreover, on the occasion different from the administration and the electroencephalogram measurement, each sample was orally ingested by the same six subjects, and the subject slapped the subject's shoulder after going to bed. As a result, it was confirmed for each subject that the subject woke up easily, that is, the sleep after each sample (the sample of Example 20 and Comparative Example 9) was orally ingested by each subject was natural sleep. .

(Electroencephalogram analysis)
The electroencephalogram was analyzed by requesting Sleepwell. The depth of sleep can be known from the delta power value of the electroencephalogram during sleep. The greater the delta power value, the deeper the sleep depth. In general, non-REM sleep appears immediately after going to bed, and REM sleep appears thereafter. In this experiment, the delta power value during non-REM sleep that appeared immediately after going to bed (that is, the delta power value until REM sleep appears after going into non-REM sleep after going to bed) was taken as the delta power value in the early stage of sleep. .

  Delta power values vary from person to person. Therefore, after ingesting the sample of Example 20 or the sample of Comparative Example 9, the delta power value in the early stage of sleep was measured four times for each subject, and the average value of the four measured values was calculated. For each subject, the change rate (%) of the delta power value after ingestion of the MTA-containing yeast (sample of Example 20) relative to the delta power value after ingestion of the placebo (sample of Comparative Example 9) is calculated by the following equation (2). did.

[Formula (2)]
Delta power value change rate (%) =
{(Average value of delta power value after sample intake of Example 20) / (Average value of delta power value after sample intake of Comparative Example 9)} × 100

  Table 5 shows the rate of change of the delta power value. The rate of change of the delta power value is an average value of six subjects.

  Table 5 shows the following. In Example 20, in which a MTA-containing yeast tablet was orally ingested as a sample, the delta power value in the early stage of sleep was larger than that in Comparative Example 9 in which a placebo tablet was orally ingested. This indicates that the yeast culture and internal use composition of the present invention can deepen non-REM sleep early in sleep, thereby promoting sleep and inducing good natural sleep. ing.

Claims (11)

Containing methylthioadenosine or a salt thereof,
The methylthioadenosine content per dry mass of the yeast culture is 5.5% by mass or more,
A sleep promoting agent comprising yeast culture as an active ingredient . S-adenosylmethionine degradation product is contained , the S-adenosylmethionine degradation product contains methylthioadenosine, and the methylthioadenosine content per dry mass of the yeast culture is 5.5% by mass or more.
A sleep promoting agent comprising yeast culture as an active ingredient . The sleep promoting agent according to claim 1 or 2, wherein the intake amount of methylthioadenosine per adult is 0.01 to 1000 mg / day. The method for producing a sleep promoter according to any one of claims 1 to 3, wherein a yeast culture containing S-adenosylmethionine or a salt thereof is heated to decompose S-adenosylmethionine or a salt thereof. Including yeast and methylthioadenosine or a salt thereof,
A / (A + B) is 1 / 10,000 or more and 1/2 or less [wherein, A represents methylthioadenosine content in the composition, and B represents yeast content in the composition, respectively. ],
Oral composition for promoting sleep . The internal use composition for sleep promotion of Claim 5 whose A / (A + B) is 1/750 or more and 1/5 or less. The internal use composition for sleep promotion of Claim 5 whose A / (A + B) is 1/100 or more and 1/5 or less. The internal use composition for sleep promotion of Claim 5 whose A / (A + B) is 5.5 / 100 or more and 1/5 or less. The composition for internal use for promoting sleep according to any one of claims 5 to 8 , wherein the amount of methylthioadenosine intake per adult is 0.01 to 1000 mg / day. The food composition for sleep promotion containing the sleep promoter of any one of Claims 1-3, and / or the internal use composition for sleep promotion of any one of Claims 5-9 . The food composition for promoting sleep according to claim 10, wherein the intake amount of methylthioadenosine per adult is 0.01 to 1000 mg / day.