JP2014065695A - Sleep-improving agent containing leucine - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an agent which can effectively improve sleep disorder with little possibility of producing side effects.SOLUTION: This invention provides a sleep-improving agent that contains leucine as an active ingredient, and also provides a sleep-improving agent further containing glycine and/or alanine.

Description

  The present invention relates to a sleep improving agent containing an amino acid, and more particularly to a sleep improving agent containing leucine.

  In recent years, with changes in social life, an increasing number of people suffer from sleep disorders due to daily stress and the like. The subjective symptoms are symptoms related to the amount of sleep, such as “short sleep time” in the past, so “bad sleep”, “slow sleep”, “wake up many times”, “bad awakening” ”,“ No feeling of deep sleep ”,“ sleepy in the daytime ”, and so on. Sleep plays an important role not only for resting the body but also for resting the cerebrum that controls the body and relieving physical and mental stress. Therefore, the disorder of sleep function causes chronic fatigue, concentration, judgment and memory deterioration, and causes various obstacles in daily life and social life.

  In order to protect the body from physical and mental stress, it is important to improve the quality of sleep in addition to ensuring sufficient sleep time. In general, the sleep of a healthy person at night depends on the sleep phase of REM sleep (shallow sleep, the body is sleeping but the brain is active) and non-REM sleep (deep sleep, other than REM sleep). It is configured. In the sleep of a healthy person, 30 minutes to 1 hour after falling asleep enters the deepest non-REM sleep, and after that, after passing through a sleep cycle that repeats shallowing and deepening at intervals of 60 to 120 minutes, awakening occurs. In addition, REM sleep during the sleep cycle becomes longer with time, and the depth of non-REM sleep tends to become gradually shallower. Such a loss of sleep balance lowers the quality of sleep, and the subjective symptoms of sleep disorders become apparent.

  To alleviate sleep disorders, in addition to improving the bedding and sleeping environment, pharmacological drugs such as benzodiazepine hypnotics, thienodiazepine hypnotics, barbituric acid hypnotics, and general-purpose drugs mainly containing diphenhydramine hydrochloride Drugs are widely used. The ethical drugs are prescribed mainly for patients diagnosed with chronic sleep disorder and have a relatively strong central depressing action. For this reason, a doctor's diagnosis is required and it is difficult to prescribe for patients with mild sleep disorders. In addition, side effects such as head sensation when waking up, nausea, headache, drug dependence, and carry-over effects may occur. On the other hand, over-the-counter drugs are sold to improve mild or temporary sleep disturbances. In particular, diphenhydramine hydrochloride, which is an antihistamine, is taken by many people who suffer from sleep disorders in order to relieve symptoms such as temporary poor sleep and light sleep. However, if a sufficient amount of an antihistamine is taken to alleviate the symptoms, the sleep effect is sustained even when waking up, and due to excessive sleepiness, a sufficient effect on the awakening at waking up cannot be expected. If the amount of antihistamine is reduced in order to reduce sleepiness when waking up, there is a problem that the awakening is further deteriorated because the poor sleep and shallow sleep cannot be sufficiently improved. For this reason, there is a demand for a sleep improving agent that has a sufficient improvement effect on sleep disorders and at the same time has no risk of causing side effects.

  So far, it has been reported that glycine has an effect of improving deep sleep disorder (Patent Document 1). Glycine is an amino acid that is recognized as a food additive and is contained in many foods. Therefore, the possibility of causing side effects is extremely low, and it can be said that the material is sufficiently safe. Since amino acids are present not only in foods but also in living bodies, it is generally considered that there is no risk of side effects compared to other drugs. Other research results on the effect of amino acid on sleep improvement have been reported. For example, branched chain amino acid improves sleep quality for patients with chronic kidney injury (Patent Document 2), sleep by intraventricular administration of proline and cysteine The effect | action etc. are mentioned (nonpatent literature 1, 2). However, on the other hand, it has been reported that histidine does not exhibit sleep action even when administered intraventricularly (Non-patent Document 3). Thus, the effect of amino acids on sleep varies depending on the type of amino acid. The degree of the effect may vary depending on the type of amino acid. Therefore, among amino acids that have no fear of side effects, a material that has a better sleep improvement effect is demanded.

Japanese Patent No. 4913410 Japanese Patent Laid-Open No. 7-324030

Hamasu K et al., Amino Acids. 2010 Jan; 38 (1): 57-64. Yamane H et al., Anim Sci J. 2009 Aug; 80 (4): 428-32. Tomonaga S et al., Brain Res Bull. 2004 Mar; 63 (1): 75-82.

  An object of this invention is to provide the agent which has little possibility of producing a side effect and can improve sleep disorder effectively.

  As a result of intensive studies to solve the above problems, the present inventors have effectively improved sleep disorders by administering leucine, which is a kind of amino acid, and leucine enhances the sleep improving action of glycine. I found out. In addition to leucine, the present inventors have found that alanine has a sleep-improving action, and that when leucine and alanine are used in combination, the sleep-improving effect is improved. Based on these findings, the present inventors have made further studies to complete the present invention.

That is, the present invention relates to the following.
[1] A sleep improving agent comprising leucine as an active ingredient.
[2] The agent according to [1] above, wherein the leucine is D-leucine.
[3] The agent according to [1] or [2], further containing glycine.
[4] The agent according to any one of [1] to [3], further containing alanine.
[5] The agent according to [4] above, wherein the alanine is D-alanine.
[6] The agent according to any one of [1] to [5], wherein the sleep improvement is an improvement of insomnia.
[7] The agent according to [6] above, wherein the insomnia is sleep onset disorder, mid-wake awakening, early morning awakening, or deep sleep disorder.
[8] The agent according to any one of [1] to [5], wherein the sleep improvement is an improvement of circadian rhythm sleep disorder.
[9] The agent according to [8] above, wherein the circadian rhythm sleep disorder is a sleep phase regression syndrome.

  ADVANTAGE OF THE INVENTION According to this invention, the sleep improving agent which can improve a sleep disorder effectively can be provided. Since the active ingredient is an amino acid, the sleep-improving agent of the present invention is excellent in safety in that it is less likely to cause side effects, and can be used on a daily basis. By improving the above sleep disorder, not only the amount of sleep such as "short sleep time", but also "bad sleep", "slow sleep", "wake up many times", "woke up poorly", "feeling of deep sleep" The quality of sleep, such as “I do n’t have it” or “I feel sleepy during the day”, can be improved.

  Further, according to the sleep improving agent of the present invention, through improvement of sleep disorder, chronic fatigue caused by sleep disorder and a decrease in concentration, judgment and memory (specifically, daytime somnolence, work Inefficiency, accidents, irritation, shortness, headache when waking up, mood modulation, decreased libido, hallucinations, hearing loss, abnormal behavior at night, etc., and society with various symptoms caused by sleep disorders Maladaptation, anxiety and depression can be prevented.

FIG. 1 is a graph showing the ratio of sleep structures 3 hours after administration of D-leucine. FIG. 2 is a diagram showing sleep time of the D-leucine group and the control group. The upper figure is a graph showing changes in sleep time every 10 minutes in both groups, where the vertical axis of the graph shows sleep time (minutes) and the horizontal axis shows time (minutes) after administration of the test substance. The lower left figure shows the sleep latency (minutes) of both groups. The lower right figure shows the sleep time (minutes) of both groups. FIG. 3 is a graph showing the effect on the self-issued amount of D-leucine. The upper figure is when D-leucine is administered at 1.75 g / kg, the lower left figure is when D-leucine is administered at 0.87 g / kg, and the lower right figure is when D-leucine is administered at 0.44 g / kg. Each case graph is shown. In any graph, the vertical axis represents the self-issued movement amount (number of times), and the horizontal axis represents the time (minutes) after administration of the test substance. FIG. 4 is a graph comparing the effects of D-leucine and L-leucine on the self-issued amount. The vertical axis of the graph shows the amount of spontaneous movement (number of times), and the horizontal axis shows the time (minutes) after administration of the test substance. FIG. 5 is a graph showing the combined effect of glycine and D-leucine for 150 minutes after administration of the test substance. The vertical axis of the graph represents the self-issued amount of movement, and the horizontal axis represents each group. FIG. 6 is a graph showing the combined effect of glycine and D-leucine every 30 minutes after test substance administration. The vertical axis of the graph represents the self-issued movement amount, and the horizontal axis represents the time every 30 minutes. FIG. 7 is a diagram showing a phase-reverse action and a phase-forward action of behavior by ingestion of a high D-leucine-containing diet according to time zones. FIG. 8 is a graph showing the effect on the self-issued amount of D-alanine. The vertical axis of the graph shows the amount of spontaneous movement (number of times), and the horizontal axis shows the time (minutes) after administration of the test substance. FIG. 9 is a graph showing the effect of D-alanine on deep body temperature. The vertical axis of the graph represents deep body temperature (° C.), and the horizontal axis represents time (minutes) before and after administration of the test substance. FIG. 10 is a graph showing the combined effect of D-alanine and D-leucine for 150 minutes after administration of the test substance. The vertical axis of the graph represents the self-issued amount of movement, and the horizontal axis represents each group.

  The present invention provides a sleep improving agent comprising leucine as an active ingredient.

  The leucine contained in the sleep-improving agent of the present invention may be any of L-form (L-leucine), D-form (D-leucine) or a mixture thereof (for example, racemate). Preferably, D-form and a mixture of D-form and L-form are used, and more preferably D-form is used.

  The sleep improving agent of the present invention preferably does not contain other branched chain amino acids (isoleucine, valine).

In the present invention, leucine may be any natural product derived from animals or plants, or one obtained by a chemical synthesis method, fermentation method, enzyme method, or gene recombination method.
L-leucine can be produced, for example, by fermentation using an L-leucine-producing bacterium belonging to the genus Escherichia, Brevibacterium, Corynebacterium or Serratia, or a mutant thereof.
D-leucine can be produced, for example, by a hydrolysis method using a D-aminoacylase derived from a bacterium belonging to the genus Alcaligenes, Pseudomonas, Streptomyces or Variovorax. Specifically, after synthesizing a racemate of N-acyl leucine, N-acyl-D-leucine is specifically hydrolyzed using D-aminoacylase, and the solubility of the mixture after hydrolysis is determined. D-leucine can be separated using a difference or the like.

  The leucine used in the present invention may be in the form of a salt as well as a free form. Examples of the salt form include acid addition salts and salts with bases, and pharmacologically acceptable salts are preferably selected. Examples of such salts include salts with inorganic acids, salts with organic acids, salts with inorganic bases, and salts with organic bases.

Examples of the salt with an inorganic acid include salts with hydrohalic acid (hydrochloric acid, hydrobromic acid, hydroiodic acid, etc.), sulfuric acid, nitric acid, phosphoric acid and the like.
Examples of the salt with an organic acid include salts with formic acid, acetic acid, propionic acid, oxalic acid, succinic acid, maleic acid, fumaric acid, citric acid and the like.
Examples of the salt with an inorganic base include a salt with an alkali metal such as sodium, potassium and lithium, a salt with an alkaline earth metal such as calcium and magnesium, and a salt with ammonium.
Examples of the salt with an organic base include salts with ethylenediamine, propylenediamine, ethanolamine, monoalkylethanolamine, dialkylethanolamine, diethanolamine, triethanolamine and the like.

  The sleep-improving agent of the present invention may be the active ingredient as it is, or, if necessary in the preparation, formulated with an appropriate pharmaceutically acceptable carrier and formulated as a pharmaceutical by a method known per se. May be. Examples of pharmaceutically acceptable carriers include excipients, binders, lubricants, solvents, disintegrants, solubilizers, suspending agents, emulsifiers, isotonic agents, stabilizers, soothing agents. Agents, preservatives, antioxidants, flavoring agents, coloring agents and the like.

  Examples of excipients include sugars such as lactose, glucose and D-mannitol, organic excipients such as starches and celluloses such as crystalline cellulose, and inorganic excipients such as calcium carbonate and kaolin. As a lubricant, pregelatinized starch, gelatin, gum arabic, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, crystalline cellulose, D-mannitol, trehalose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, etc. Is stearic acid, fatty acid salts such as stearate, talc, silicates, etc., solvent is purified water, physiological saline, etc., disintegrator is low substituted hydroxypropyl cellulose, chemically modified Cellulose, starch, etc., as solubilizers, polyethylene glycol, propylene glycol, trehalose, benzyl benzoate, ethanol, sodium carbonate, sodium citrate, sodium salicylate, sodium acetate, etc. are used as suspending agents or emulsifiers. Examples of isotonic agents include sodium lauryl sulfate, gum arabic, gelatin, lecithin, glyceryl monostearate, polyvinyl alcohol, polyvinyl pyrrolidone, sodium carboxymethyl cellulose, polysorbates, polyoxyethylene hydrogenated castor oil, etc. , Sodium chloride, potassium chloride, saccharides, glycerin, urea, etc., stabilizers include polyethylene glycol, sodium dextran sulfate, and other amino acids However, as a soothing agent, glucose, calcium gluconate, procaine hydrochloride, etc., and as preservatives, paraoxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, etc. are antioxidants. As sulfites, ascorbic acid and the like, as flavoring agents, sweeteners and fragrances commonly used in the pharmaceutical field, and as coloring agents, colorants commonly used in the pharmaceutical field are mentioned. .

  The dosage form of the sleep improving agent of the present invention formulated is not particularly limited, but for oral administration, for example, solids such as powders, granules, capsules, tablets, chewables, solutions, syrups, etc. It can be set as a liquid agent. In addition, the above dosage forms can be used for parenteral administration, for example, injections, infusions, nasal / pulmonary sprays, and the like.

  The sleep improving agent of the present invention is also useful as a food. In this specification, “food” means food in general (including beverages), special-purpose foods (foods for sick persons, foods for specified health use) and diets specified in the Special-Use Food System of the Consumer Affairs Agency. Tally supplements are also included.

  When the sleep-improving agent of the present invention is used as food, the sleep-improving agent of the present invention may be an active ingredient as it is, but it is formulated by a method known per se using an appropriate food-acceptable carrier. May be. Examples of the formulated dosage form of the sleep improving agent of the present invention include powders, tablets, capsules, drinks and the like, but are not particularly limited thereto. Examples of the carrier acceptable as food include processing ingredients, seasonings, flavorings, sweeteners, and the like, and the pharmaceutically acceptable carriers described above are also included. Moreover, vitamins, other nutritional supplements, and the like can be blended in the formulation.

  The content of leucine in the sleep-improving agent of the present invention when formulated is not particularly limited, and can be appropriately adjusted so as to contain an amount necessary for producing the effects of the present invention. The content of leucine varies depending on the type of dosage form and the type of carrier used, but is, for example, 0.015 to 100% by weight, preferably 0.02 to 99% by weight, based on the total weight of the preparation. Yes, more preferably 0.025 to 98.5% by weight, still more preferably 0.03 to 98% by weight. In addition, when leucine forms a salt, calculation of content shall be performed after converting the salt into a free body.

  According to the sleep improving agent of the present invention, sleep disorders can be improved. As used herein, “sleep disorder” refers to a state in which there is some abnormality in sleep or sleep, and includes a decrease in the amount of sleep and a decrease in sleep quality. Symptoms of a decrease in the amount of sleep include, for example, an increase in the time until bedtime, insufficient sleep time due to mid-wake or early morning awakening, and the like. In addition, examples of the symptoms of sleep quality deterioration include a shift in sleep time, a decrease in deep sleep (non-REM sleep), a decrease in REM sleep, a break in sleep due to mid-wake awakening, and a nap in an active time period. . In the diagnosis of sleep disorder, a test composed of a plurality of question items is usually used. For definitive diagnosis, electroencephalogram measurement or polysomnography that measures a plurality of items including electroencephalogram measurement can be used. Diagnostic results can be classified according to internationally accepted standards (The International Classification of Sleep Disorder, ICSD). In the present specification, “improvement” is meant to include the repair, suppression (delay of progression), mitigation, regulation and prevention (including both prevention of occurrence and recurrence) of sleep disorders and symptoms caused thereby. used.

  In the present invention, sleep disorders include sleep abnormalities, circadian rhythm sleep disorders, sleep-related comorbidities, sleep disorders related to mental disorders, sleep disorders related to physical diseases, sleep disorders related to drugs, etc. It is.

  Sleep abnormalities encompass the concepts of insomnia and hypersomnia. Insomnia refers to any complaint of sleep disturbance, mid-wake awakening, early morning awakening, or deep sleep disorder. Hypersomnia refers to a state in which excessive sleepiness during the day or a state in which it actually falls asleep is repeatedly observed. In either case, the duration is at least one month, but is not particularly limited thereto. Specific examples of sleep disorders include psychophysiological insomnia, primary hypersomnia, narcolepsy, sleep apnea syndrome, repetitive hypersomnia, idiopathic hypersomnia, periodic limb movement disorders, restless leg syndrome, Examples include time zone change syndrome and environmental sleep disorder.

  The circadian rhythm sleep disorder does not match the rhythm of the day and night cycle with the rhythm of the body clock. It is a sleep disorder that causes difficulty in activities. Circadian rhythm sleep disorders include sleep phase regression syndrome, which is difficult to get up early in the morning without sleeping until dawn, and sleep phase advance syndrome, which is sleepy early in the evening and awakens early in the morning.

  Parasomnia is a general term for psychosomatic abnormalities that occur during the course of sleep. Specific examples thereof include, but are not limited to, sleepwalking, night wonder, REM sleep behavior disorder, sleep paralysis, etc. Can be mentioned.

  Examples of diseases that cause sleep disorders related to mental disorders include depression, mania, and schizophrenia. Examples of the diseases that cause sleep disorders related to physical diseases include Alzheimer type dementia, cerebrovascular dementia, and Parkinson's disease. Examples of sleep disorders associated with drugs include sleep disorders induced by anticonvulsants, anxiolytics, antipsychotics, psychostimulants, antiparkinsonian drugs, anticancer drugs, hormonal agents and the like.

  The sleep-improving agent of the present invention significantly increases the ratio of the non-REM sleep period and the non-REM sleep time, and thus is particularly excellent in the effect of maintaining stable sleep. As a result, the sleep-improving agent of the present invention can effectively improve not only the amount of sleep but also the quality of sleep in terms of deep sleep, a reduction in the number of times awakened, and a sufficient feeling of deep sleep. Therefore, it is preferable to apply the sleep improving agent of the present invention to insomnia, and among them, it is particularly preferable to apply it to mid-wake awakening, early morning awakening, and deep sleep disorder.

  Moreover, since the sleep improving agent of this invention reduces the ratio of an awakening period or shortens sleep latency, it can improve the quality of sleep in terms of improving sleep. Therefore, the sleep-improving agent of the present invention is preferably applied not only to the above-mentioned arousal, early-morning awakening, and deep sleep disorder, but also to sleep disorder.

  Furthermore, since the sleep improving agent of the present invention has a circadian rhythm regulating action, it is preferably applied to circadian rhythm sleep disorders. In particular, since the sleep improving agent of the present invention contributes to the advance of circadian rhythm, it is useful for sleep phase regression syndrome among circadian rhythm sleep disorders. Through the adjustment of circadian rhythm, it is possible to improve time symptom (time difference blur), improvement of poor physical condition and visual loss associated with shift work, and adjustment of the body clock in the case of visual impairment.

  Mammals (for example, humans, mice, rats, hamsters, rabbits, cats, dogs, cows, sheep, monkeys, etc.) can be used as the application target of the sleep improving agent of the present invention. The application target of the sleep improving agent of the present invention is preferably a human. In addition, when adapting to mammals other than a human, the dosage of the sleep improving agent of this invention should just be adjusted suitably according to the body weight or magnitude | size of an animal.

  The administration method when the sleep improving agent of the present invention is used as a pharmaceutical may be either oral administration or parenteral administration. From the viewpoint of reducing the burden on the subject, the sleep improving agent of the present invention is preferably administered orally to the subject. On the other hand, for subjects who are difficult to administer orally, intravenous or transarterial administration can be performed as an infusion solution.

  The daily dose (intake) of the sleep-improving agent of the present invention for adults varies depending on the age, body weight, symptom, administration method and the like of the subject patient, but is usually 0.000625 as the weight of leucine as an active ingredient. 2.0 g / kg / day (daily intake per kg body weight), preferably 0.0007 to 1.5 g / kg / day, more preferably 0.00075 to 1.0 g / kg / day, more preferably 0.0008 to 0.5 g / kg / day. When leucine forms a salt, the dose should be calculated after converting the salt into a free form.

  When calculating the dosage of the amino acid that is the active ingredient used in the present invention, the above calculation range is determined as the active ingredient of the drug used for improving sleep which is the object of the present invention. It is not necessary to include in the calculation for amino acids that are ingested or administered for other purposes, such as for the need of a normal diet or for the treatment of another disease. For example, it is not necessary to calculate by deducting the amount of amino acids per day taken from a normal diet from the daily dose of the active ingredient in the present invention.

  In the present invention, the above-mentioned daily dose is administered once or divided into 2 to several times as necessary, preferably twice in the morning and evening, and administered at appropriate intervals. The timing of administration is not particularly limited, and examples thereof include when waking up, before going to bed, before meals, between meals, after meals, etc., preferably when getting up and before going to bed. As described above, since the sleep-improving agent of the present invention can advance the circadian rhythm, it is administered at the early morning or at the start of activities such as when waking up, at breakfast (before meals, between meals, after meals), before work at shift work, etc. It is also possible to promote sleep at night effectively.

  The administration period of the sleep-improving agent of the present invention is not particularly limited as long as the effect of the present invention can be obtained. When the sleep effect immediately after administration is desired, it is usually 1 day or more, preferably 3 days or more. In that case, the sleep-improving agent of the present invention is usually administered every day, preferably when it is difficult to fall asleep. On the other hand, when the forward effect of circadian rhythm is desired, the administration period of the sleep improving agent of the present invention is usually 1 day or more, preferably 3 days or more, more preferably 4 days or more. In that case, the sleep-improving agent of the present invention is usually administered during a time difference symptom period or a shift work period, preferably within 6 hours from waking up during that period, and more preferably when waking up during that period.

  The sleep improving agent of the present invention can further contain glycine. Glycine has the effect | action which improves a deep sleep disorder as mentioned above, and there exists an effect which improves the quality of sleep. Leucine contained in the sleep improving agent of the present invention has an action of enhancing the sleep improving effect of such glycine.

  In the present invention, any glycine derived from animals or plants, or those obtained by chemical synthesis, fermentation, enzyme, or gene recombination may be used. The glycine used in the present invention may be in the form of a salt as well as a free form. Although it does not specifically limit as a form of this salt, What was illustrated above is mentioned.

  In the sleep improving agent of the present invention, leucine and glycine may be used in combination. Here, the term “in combination” as used herein means the use of another active ingredient before, simultaneously with, or after the administration of one active ingredient, and as a compounding agent in which each active ingredient is mixed. It is meant to include use. In the present invention, as long as both leucine and glycine are used and the sleep improving effect of glycine is enhanced, their dosage forms are not limited.

  Examples of such administration forms include (1) administration of a single preparation obtained by simultaneously formulating leucine and glycine, and (2) two preparations obtained by separately formulating leucine and glycine. Simultaneous administration by the same route of administration, (3) administration of two types of preparations obtained by separately formulating leucine and glycine with a time difference in the same route of administration, and (4) leucine and glycine separately Simultaneous administration of two preparations obtained by formulation into different administration routes, (5) Administration of two preparations obtained by separately formulating leucine and glycine at different time intervals in different administration routes (For example, administration in which leucine is first administered and then glycine is administered, or vice versa). In the present invention, the administration form (1) or (2) is preferred, and the administration form (1) is more preferred, from the viewpoint of omitting the number of administrations and administration labor.

  The content of glycine in the sleep-improving agent of the present invention is not particularly limited, and can be appropriately adjusted so as to contain an amount necessary for producing the effects of the present invention. When glycine forms a salt, the content is calculated after converting the salt into a free form.

  The ratio (weight ratio) of leucine and glycine contained in the sleep-improving agent of the present invention can be appropriately adjusted within the range having the effects of the present invention. For example, the ratio of both amino acids (leucine: glycine) is usually 0.005: 1.0 to 1.0: 1.0, preferably 0.01: 1.0 to 0.8: as a weight ratio. 1.0, more preferably 0.02: 1.0 to 0.6: 1.0. When leucine and glycine form a salt, the weight ratio is calculated after converting each amino acid salt into a free form. Here, in this specification, "weight ratio" shows the ratio of the weight of each amino acid contained in the sleep improving agent of this invention, for example, when two types of amino acids are included in one composition. It is the ratio of the content of individual amino acids, and when each of the two amino acids is separately included in the two compositions, it is the ratio of the weight of each amino acid contained in each composition.

In the present invention, the dose (intake amount) of glycine, the administration period, and the administration interval can be appropriately set according to the subject patient, symptoms and the like. The dose of glycine is not particularly limited, but is usually 0.00208 to 2.5 g / kg / day (intake per day per kg of body weight), preferably 0.0025 to 2.0 g / kg / day. day, more preferably 0.0125 to 1.5 g / kg / day. When glycine forms a salt, the above dose is calculated after converting the salt into a free form. The administration period and administration interval of glycine are not particularly limited, and can be combined with, for example, the administration period and administration interval of the sleep improving agent of the present invention.
Similar to the above leucine, when calculating the dosage of glycine, the above calculation range is determined as the active ingredient of the drug used for improving sleep which is the object of the present invention. It is not necessary to include this in the calculation for glycine that is ingested or administered for that purpose, for example from the need for a normal diet or for the treatment of another disease.

  As described above, when administering the sleep improving agent of the present invention, leucine and glycine may be administered at the same time, leucine may be administered first, glycine may be administered, or glycine may be administered first. May be administered followed by leucine.

  The sleep improving agent of the present invention can further contain alanine. Alanine has the effect | action which raises the sleep improvement effect of leucine. Therefore, the combined use of alanine and leucine is one preferred embodiment in the present invention. Further, as described above, leucine enhances the sleep-improving action of glycine. Therefore, using glycine in combination with alanine and leucine is another preferred embodiment of the present invention. In addition, as shown in the below-mentioned Example, alanine itself has a sleep improvement effect. Therefore, alanine can be used alone as an active ingredient.

  The alanine used in the present invention may be any of L-form (L-alanine), D-form (D-alanine) or a mixture thereof (for example, racemate), but in the present invention, preferably D-form. And a mixture of D-form and L-form, and more preferably D-form is used.

In the present invention, alanine may be any natural one derived from animals or plants, or one obtained by a chemical synthesis method, fermentation method, enzyme method, or gene recombination method.
L-alanine can be produced, for example, by enzymatic conversion of L-aspartate using aspartate decarboxylase or a bacterium having the enzyme activity (genus Xanthomonas).
D-alanine is produced, for example, by fermentation using D-alanine-producing bacteria belonging to the genus Brevibacterium or Corynebacterium or mutants thereof, or by enzymatic conversion of L-alanine using alanine racemase. Can do.

  In addition, alanine used in the present invention may be in the form of a salt as well as a free form. Although it does not specifically limit as a form of this salt, What was illustrated above is mentioned.

  When alanine is used in the present invention, the sleep-improving agent of the present invention has the effect of lowering the deep body temperature in addition to the effects described above, such as deep sleep, reduction in the number of times awakened, sufficient deep sleep feeling, etc. Helps improve sleep quality. Therefore, even when alanine is used, the sleep-improving agent of the present invention is preferably applied to insomnia, and more preferably applied to midway awakening, early morning awakening, and deep sleep disorder.

  Alanine can be used in combination with leucine and glycine, and the dosage form is not particularly limited as long as the effects of the present invention are exhibited. Alanine is particularly preferably used in combination with leucine because it increases the sleep-improving effect of leucine.

  When alanine is used in combination with leucine, for example, (1) administration of a single preparation obtained by simultaneously formulating leucine and alanine, and (2) formulating leucine and alanine separately Simultaneous administration of the two obtained preparations by the same administration route, (3) administration of the two preparations obtained by separately formulating leucine and alanine with a time difference in the same administration route, (4) Simultaneous administration of two preparations obtained by separately formulating leucine and alanine in different administration routes, (5) Different administration routes of two preparations obtained by separately formulating leucine and alanine Administration at a time difference (for example, administration in which leucine is administered first and then alanine is administered, or vice versa). In the present invention, the administration form (1) or (2) is preferred, and the administration form (1) is more preferred, from the viewpoint of omitting the number of administrations and administration labor.

  The content of alanine in the sleep-improving agent of the present invention is not particularly limited, and can be appropriately adjusted so as to contain an amount necessary to achieve the effects of the present invention. In addition, when alanine forms a salt, content calculation shall be performed after converting the salt into a free form.

  The ratio (weight ratio) of leucine and alanine contained in the sleep-improving agent of the present invention can be appropriately adjusted within the range having the effects of the present invention. For example, the ratio of both amino acids (leucine: alanine) is usually 0.005: 1.0 to 1.0: 1.0, preferably 0.01: 1.0 to 0.8: as a weight ratio. 1.0, more preferably 0.02: 1.0 to 0.6: 1.0. When leucine and alanine form a salt, the weight ratio is calculated after converting each amino acid salt into a free form.

  In the present invention, the dose (intake amount) of alanine, the administration period, and the administration interval can be appropriately set according to the subject patient, symptoms and the like. The dose of alanine is not particularly limited, but is usually 0.00208 to 2.5 g / kg / day (intake per day per kg of body weight), preferably 0.00625 to 2.0 g / kg / day. day, more preferably 0.0125 to 1.5 g / kg / day. When alanine forms a salt, the above dose is calculated after converting the salt into a free form. The administration period and administration interval of alanine are not particularly limited, and can be combined with, for example, the administration period and administration interval of the sleep improving agent of the present invention.

  As described above, when administering the sleep improving agent of the present invention, leucine and alanine may be administered at the same time, leucine may be administered first, then alanine may be administered, or alanine may be administered first. May be administered followed by leucine.

  In the present invention, when three amino acids of leucine, glycine and alanine are used, the administration form is not particularly limited. For example, (1) a single preparation obtained by simultaneously formulating the above three amino acids. Administration, (2) simultaneous administration by the same route of three preparations obtained by separately formulating the above three amino acids, (3) three preparations obtained by separately formulating the above three amino acids (4) Simultaneous administration by different administration routes of three preparations obtained by separately formulating the three kinds of amino acids, (5) Three kinds of amino acids And administration of the three preparations obtained by separately formulating at different time intervals in different administration routes. In the present invention, the administration form (1) or (2) is preferred, and the administration form (1) is more preferred, from the viewpoint of omitting the number of administrations and administration labor. In addition, any two of the above three amino acids may be formulated at the same time, and such a formulation and a formulation obtained by formulating the remaining one amino acid are routed in the same or different route. At the same time or at different times. Regarding the above three amino acids, the order of amino acids to be administered is not particularly limited, and even when two arbitrary amino acids are formulated, the formulation and the remaining one amino acid are administered. The order is not particularly limited.

  The ratio (weight ratio) of leucine, glycine and alanine in the sleep-improving agent of the present invention can be appropriately adjusted within the range having the effects of the present invention. For example, the ratio of the above three amino acids (leucine: glycine: alanine) is usually 0.005: 0.5: 0.005-1.0: 1.0: 1.0 as a weight ratio, preferably Is 0.01: 0.5: 0.05 to 0.8: 1.0: 1.0, more preferably 0.02: 0.5: 0.5 to 0.6: 1.0: 1.0. When the above three amino acids form a salt, the weight ratio is calculated after converting each amino acid salt into a free form.

  As described above, the three amino acids leucine, glycine and alanine may be administered at the same time, or may be administered with a time difference. Even when two arbitrary amino acids among the above three amino acids are formulated, the administration interval with the remaining one amino acid is not particularly limited, and can be the same as described above.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited to a following example.

Example 1
The sleep improvement effect of D-leucine was evaluated using a rat sleep disorder model established by Shinomiya et al. (Psychopharmacol. Berl., 173: 1-2, 203, 2004).

<Measurement method>
Male Wistar rats (200-300g) are operated under anesthesia, an electroencephalogram telemetry transmitter (TL11M2-F40-EET: DSI) is inserted into the abdominal cavity, and an electroencephalogram acquisition electrode is connected to the head and myoelectric potential. An acquisition electrode was embedded in the neck. After the operation, a recovery period of 10 days was set up, and all animals were allowed to eat freely and drink freely.
From the beginning of the light period, the rest period, the stress load was changed to a sleep disorder state for 3 hours, and sleep electroencephalograms were measured for these rats for 3 hours after administration of the test substance. The stress load and sleep electroencephalogram were measured according to the method of Shinomiya et al. Specifically, place a wire mesh as a scaffold in the cage, and create a state where the rat is stressed by placing water so that the water surface is 1 cm below this wire mesh. The electroencephalogram and electromyogram data were measured 3 hours after administration.

<Data analysis>
The electroencephalogram and electromyogram were transmitted wirelessly from the telemetry transmitter to the receiving board, and were captured from the receiving board into the internal software (Dataquest) in the PC via the data conversion matrix, and the waveform was observed. The sleep-wake cycle is automatically analyzed by computer based on the captured electroencephalogram, electromyogram and action amount, and finally confirmed visually, and each epoch is classified as one of wake, non-REM sleep, or REM sleep did. In the awake period, when theta wave (4-8Hz) activity is observed in high amplitude electromyogram and low amplitude electroencephalogram, non-REM sleep has delta wave (1-4Hz) activity in low amplitude electromyogram and high amplitude electroencephalogram. In the case of the center, REM sleep was determined as a case where theta activity was a center in a low amplitude electromyogram and a low amplitude brain wave.
For the evaluation of sleep time, the proportion of time in the awakening period, non-REM sleep period, and REM sleep period in 3 hours from the start of measurement was calculated and compared.
The sleep latency was evaluated as the time when the first non-REM sleep that appeared for 60 seconds or more after the measurement started.

<Test substance>
A control group of rats was given a 5% by weight carboxymethylcellulose aqueous solution, and a D-leucine group of rats was given a sample administered to the control group with 0.87 g of D-leucine added per kg body weight. Administration was carried out using an oral sonde.

<Result>
FIG. 1 shows the ratios of the arousal period, non-REM sleep period, and REM sleep period 3 hours after administration of the test substance under sleep disorder conditions. Compared to the control group, the D-leucine group significantly decreased the rate of wakefulness and increased the rate of non-REM sleep. That is, it was confirmed that sleep was significantly improved by administration of D-leucine.
In addition, FIG. 2 shows non-REM sleep time change every 10 minutes, sleep latency, and non-REM sleep time 90 to 140 minutes after administration in 3 hours after administration of the test substance under sleep disorder conditions. In the D-leucine group, the non-REM sleep time increased significantly at 100, 110, and 130 minutes after administration compared to the control group, and the sleep latency tended to decrease. In the D-leucine group, the amount of non-REM sleep 90 to 140 minutes after administration was significantly increased. From these results, it was confirmed that administration of D-leucine improves sleep quality, that is, improves sleep and provides stable sleep.

(Example 2)
Since the self-issued amount of movement decreases during sleep, the numerical value indicating the self-issued amount of movement is used as an indirect index for evaluating sleep. Therefore, in order to evaluate the concentration-dependent effect of D-leucine, the amount of spontaneous movement was measured at three concentrations of 1.75 g / kg, 0.87 g / kg, and 0.44 g / kg.

<Measurement method>
Using male C57Bl / 6J mice aged 9 to 11 weeks, self-issued dynamics were evaluated with a self-issued dynamics measuring device (MELQUEST, MV-40) 2 hours after the start of the light period.

<Test substance>
Control group mice were given 5% by weight carboxymethylcellulose aqueous solution, and D-leucine group mice were added to the control group with 1.75 g, 0.87 g, and 0.44 g D-leucine per kg body weight. Was administered. Administration was carried out using an oral sonde.

<Result>
The above results are shown in FIG. D-Leucine (1.75 g / kg) showed the largest self-issued movement reduction effect, followed by 0.87 g / kg and 0.44 g / kg in order of large self-issued movement reduction effect. No significant difference was observed at 0.44 g / kg. Thus, it was confirmed that D-leucine has a concentration-dependent sleep improving effect.

(Example 3)
The difference in the activity of lowering the self-issued amount was evaluated for the optical isomers of leucine, D-leucine and L-leucine.

<Measurement method>
Same as Example 2 above.

<Test substance>
Control mice received 5 wt% carboxymethylcellulose aqueous solution, and mice in the D-leucine and L-leucine groups had 1.75 g D-leucine and L-leucine per kg body weight in the samples administered to the control group. Each of which was added was administered. Administration was carried out using an oral sonde.

<Result>
FIG. 4 shows the result of comparison of the action of lowering the amount of spontaneous movement in the D-leucine group and the L-leucine group. The D-leucine group showed a significant decrease in the amount of spontaneous movement compared to the control group. In the L-leucine group, although the difference from the control group was not as great as in the D-leucine group, the amount of spontaneous movement decreased significantly in some time zones. From these results, it was considered that both D-leucine and L-leucine have sleep-improving action.

Example 4
The self-issued amount-lowering effect by simultaneous administration of glycine and D-leucine, which have sleep-improving action, was evaluated.

<Measurement method>
Same as Example 2 above.

<Test substance>
A control group of mice was administered with a 5% by weight carboxymethylcellulose aqueous solution, and the other group of mice was administered with a sample to which the following amounts of glycine and D-leucine had been added. Administration was carried out using an oral sonde.
Glycine (1.5g) group (Glycine: 1.5g / kg)
Glycine (1g) group (Glycine: 1g / kg)
D-leucine (0.5g) group (D-leucine: 0.5g / kg)
Glycine (1g) + D-leucine (0.5g) group (Glycine: 1g / kg, D-leucine: 0.5g / kg)

<Result>
The above results are shown in FIGS. There was no significant difference between the glycine (1.5 g), glycine (1 g), and D-leucine (0.5 g) groups in the spontaneous issuance in 150 minutes after administration. In the glycine (1 g) + D-leucine (0.5 g) group to which (1 g / kg) and D-leucine (0.5 g / kg) were co-administered, a significant decrease in the amount of spontaneous movement was observed. In addition, in the glycine (1 g) + D-leucine (0.5 g) group, the spontaneous movement amount was significantly reduced 30 to 60 minutes after administration and 60 to 90 minutes after administration, and the difference was in the glycine (1.5 g) group. Was significantly larger than. From these results, it was considered that D-leucine has the effect of enhancing the sleep-improving action of glycine.

(Example 5)
The phase-retarding and phase-advancing effects of D-leucine on mice reared under dark conditions for 24 hours were evaluated.
When mice are raised under dark conditions for 24 hours, external information (light-dark cycle of light) cannot be obtained, so the mice become active in the cycle specific to the biological clock (free run (in the sense that they are free from synchronization) Also called free continuation)). When this free continuation period is converted to 24 hours and shown on the horizontal axis, and the circadian time is taken, the first 12 hours are the subjective light period (the time zone when the biological clock judges it to be noon: CT0-12) The latter half is referred to as the subjective dark period (time zone that is judged to be night by the biological clock: CT12-24).
Under such conditions, the amount of spontaneous movement of the mouse was measured using an emission infrared detection type self-issued movement sensor (Neuroscience, NS-AS01). In addition, a diet containing 6 mg of D-leucine per gram of food was prepared, and replaced with normal diet for 4 hours per day. Ingestion time is CT2-6 (first half of subjective light period), CT8-12 (second half of subjective light period), CT14-18 (first half of subjective dark period), CT20-24 (second half of subjective dark period) ) Divided into four time zones. The phase change of the behavior pattern was determined after such a bait replacement for 4 days.
FIG. 7 shows the phase shift of the behavior start time of the mouse when the determination reference date is set 4 days after the final intake date of the D-leucine-rich diet. As a result, it was clarified that the effect of D-leucine was not observed in the subjective light period, but the circadian rhythm was advanced by ingestion in the second half of the subjective dark period.

  From the results of Examples 1 to 5 above, it has been clarified that D-leucine and L-leucine have circadian rhythm regulation, increase in sleep amount, sleep quality improvement and sleep-stimulating action. It was confirmed to be effective in improving sleep disorders including sleep disorders. As a result, D-leucine and L-leucine can eliminate circadian rhythm sleep disorders such as jet lag and poor physical condition associated with shift work, and sleep dissatisfaction such as light sleep, poor awakening, and poor sleep. It was found to show an excellent effect.

(Example 6)
Using the self-issued amount, which is an indirect indicator of sleep state, we evaluated the effect of D-alanine on reducing the self-issued amount.

<Measurement method>
Using male C57Bl / 6J mice aged 9 to 11 weeks, the amount of behavior was evaluated with a self-issued dynamic measurement device (MELQUEST, MV-40) 2 hours after the start of the light period.

<Test substance>
A control group of mice was administered with a 5 wt% carboxymethylcellulose aqueous solution, and a D-alanine group of mice was administered a sample administered with the control group with 1.19 g of D-alanine added per kg body weight. Administration was carried out using an oral sonde.

<Result>
The results are shown in FIG. As shown in FIG. 8, there was a tendency that the amount of spontaneous movement of mice decreased by administration of D-alanine. Thereby, it was confirmed that D-alanine has a sleep improvement effect.

(Example 7)
It is known that the deep body temperature decreases during sleep, and the numerical value indicating the deep body temperature is used as an indirect index for evaluating sleep. Therefore, the effect of D-alanine (1.19 g / kg) on deep body temperature was evaluated.

<Measurement method>
A 9-week-old male C57Bl / 6J mouse was operated under anesthesia, and a telemetry transmitter for deep body temperature measurement (TA10TA-F20: DSI) was implanted in the abdominal cavity. After the operation, a recovery period of 10 days was set up, and all animals were allowed to eat freely and drink freely.

<Data analysis>
The deep body temperature was wirelessly transmitted from the telemetry transmitter to the receiving board, and was taken into the internal software (Dataquest) in the PC via the data conversion matrix from the receiving board, and the deep body temperature was obtained every 5 minutes. The evaluation time was 60 minutes before the start of measurement and 150 minutes after administration.

<Test substance>
A control group of mice was administered with a 5 wt% carboxymethylcellulose aqueous solution, and a D-alanine group of mice was administered a sample administered with the control group with 1.19 g of D-alanine added per kg body weight. Administration was carried out using an oral sonde.

  The above results are shown in FIG. As a result, the D-alanine group showed a tendency for the deep body temperature of mice to decrease. From these results, it was suggested that D-alanine has a sleep improving action.

(Example 8)
As described above, the results of Examples 1 to 5 indicate that D-leucine has a sleep improving action. Therefore, in order to examine the combined use effect of D-leucine and D-alanine, the self-issued amount reduction effect by simultaneous administration of both was evaluated.

<Measurement method>
Same as Example 6 above.

<Test substance>
A control group of mice was administered with a 5% by weight carboxymethylcellulose aqueous solution, and the other groups of mice were administered with samples to which the following amounts of D-leucine and D-alanine had been added, respectively. Administration was carried out using an oral sonde.
D-alanine (1.5g) group (D-alanine: 1.5g / kg)
D-leucine (0.5g) group (D-leucine: 0.5g / kg)
D-alanine (1g) + D-leucine (0.5g) group (D-alanine: 1g / kg, D-leucine: 0.5g / kg)

<Result>
The above results are shown in FIG. As a result, when D-alanine and D-leucine were administered in combination, the amount of spontaneous movement tended to be lower than when D-alanine or D-leucine was administered alone. From these results, it was suggested that D-alanine has an effect of enhancing the sleep-improving action of D-leucine. Since the effect of D-leucine is enhanced in this way, it was considered that D-alanine has an effect of enhancing the effect of other D-amino acids.

  From the results of Examples 6 to 8 above, D-alanine is considered to have a sleep improving action because it decreases the spontaneous movement amount and deep body temperature, and is useful for improving sleep disorders. Was suggested. Furthermore, the effect of D-leucine was shown to be enhanced by administration in combination with D-leucine, which was confirmed to have a sleep improvement effect. D-alanine has been found to be particularly effective in resolving sleep complaints such as light sleep, poor awakening, and poor sleep.

  The sleep-improving agent of the present invention can effectively improve sleep disorders, and since the active ingredient is an amino acid, it is excellent in terms of safety. Therefore, the sleep improving agent of the present invention can be used in the pharmaceutical field. In addition, since the amino acid is contained in many foods and can be taken orally, the sleep-improving agent of the present invention is also useful in the food field.

Claims (9)

  A sleep improving agent comprising leucine as an active ingredient.   The agent according to claim 1, wherein the leucine is D-leucine.   The agent according to claim 1 or 2, further comprising glycine.   The agent according to any one of claims 1 to 3, further comprising alanine.   The agent of Claim 4 whose alanine is D-alanine.   The agent of any one of Claims 1-5 whose sleep improvement is improvement of insomnia.   The agent according to claim 6, wherein the insomnia is a sleep onset disorder, mid-wake awakening, early morning awakening or a deep sleep disorder.   The agent according to any one of claims 1 to 5, wherein the sleep improvement is an improvement of circadian rhythm sleep disorder.   The agent according to claim 8, wherein the circadian rhythm sleep disorder is a sleep phase regression syndrome.