JP6570052B2 - Circadian rhythm improving agent - Google Patents

Description

  The present invention relates to a circadian rhythm improving agent useful for improving sleep disorders and the like.

Occurrence of various forms of sleep disorders associated with aging and 24-hour social activities has become a social problem. At present, one out of every five Japanese people is said to have some problems with sleep, and in particular elderly people, one in three people are said to have trouble with sleep.
Sleep disorders are classified into sleep breathing disorders such as sleep apnea, and circadian rhythm sleep disorders (circadian rhythm sleep disorders) that are related to the onset of the circadian clock. For circadian rhythm sleep disorders, extrinsic acute syndrome caused by jet lag, night shift / shift work, shift phase sleep syndrome (DSPS), sleep phase advance syndrome (ASPS), non-24-hour sleep It can be classified into intrinsic chronic syndromes such as arousal disorder and irregular sleep-wake pattern.
As a cause of circadian rhythm sleep disorders, some familial genetic causes have been reported in advanced sleep phase syndrome, but most of them are night shifts, shift work, jet lag, irregular diet, mental It is thought that lifestyle habits such as social stress are involved. The onset of these sleep disorders is thought to be related to the body clock composed of clock genes, but the detailed mechanism is unknown.

  On the other hand, it is known that various physiological rhythms such as deep body temperature and secretory rhythms of hormones such as melatonin related to sleep decrease with age, and sleep disorders associated with aging. Is considered to involve the decay of the circadian clock (Non-Patent Document 1). Lifestyle-related diseases such as diabetes, obesity, and metabolic syndrome are known to cause sleep disorders, and it is thought that decay of the body clock is also involved in such sleep disorders. (Non-Patent Document 2).

As a treatment method for sleep disorders, high-intensity phototherapy requiring a special device, and administration of vitamin B12 (Non-patent Document 1) or melatonin preparation (Patent Document 1) are generally used. However, all of these treatment methods attempt to normalize biological rhythms by regulating abnormalities in the biological clock, and can be fundamental treatments for disorders caused by abnormal biological clocks. Absent.
In addition, benzodiazepine drugs (Non-patent Document 3) are generally used as sleeping drugs, and various half-life drugs have been developed from long-time types to short-time types, intermediate types, and long-term types. . However, the treatment of sleep disorders using these sleeping drugs is coping therapy, and it is often difficult to fundamentally treat sleep disorders. In addition, there are many side effects such as sleepwalking symptoms, and the usage thereof requires careful attention.

Recently, a 4-oxoimidazolidine-2-spiropiperidine derivative having an action as an agonist of a nociceptin receptor is expected to have a circadian rhythm sleep disorder therapeutic effect (Patent Document 2), but is still under research and development. Side effects are also a concern.
Research and development of sleep-improving agents using raw materials for various foods and beverages and natural ingredients contained in them are also thriving, using tea leaf-derived theanine (Patent Document 3), and whey such as fermented whey having an endogenous melatonin secretion effect Sleep improvers using ginseng extract (Non-patent Document 4), phosphatidylcholine (Non-patent Document 5) contained in salmon roe oil, etc., in addition to those added with a kind (Patent Document 4) . These are all derived from ingredients used in foods and beverages, luxury goods, etc., so they are highly safe and can be taken on a daily basis. The elucidation is not enough, and the individual differences in the effects are large.

  The circadian rhythm is controlled by a circadian clock, and in recent years it has become clear that the rhythm of the circadian clock is engraved by a series of genes called clock genes. The circadian clock has three functions: circadian rhythm phase adjustment, cycle length adjustment, and amplitude adjustment. In particular, the circadian rhythm amplitude and cycle length adjustment functions have various functions including sleep disorders. It is considered to be most important in the prevention / improvement and treatment of biological rhythm-related diseases. Improving various circadian rhythm decay and cycle disturbance caused by metabolic diseases such as aging, diabetes and obesity is an effective means to normalize various biological rhythms including sleep-wake rhythm Although it is expected, there has been no report on an effective sleep improving agent that is derived from a natural component with established safety and that regulates the amplitude and cycle length of circadian rhythm.

  On the other hand, in recent molecular biological studies of biological clocks centering on clock genes, reporter plasmids that link the luciferase gene to the promoters of clock genes such as Period1 (Per1), Period2 (Per2), and Bmal1 are used as fibers such as NIH3T3. Many experiments have been used in which the effect on the circadian clock is examined by introducing it into blast cells and monitoring the circadian rhythm of luciferase activity (Non-patent Documents 6 and 7). In addition, a study has been reported in which cell lines that stably express these reporter plasmids are established to examine the same action (Patent Document 5). However, in experiments using these fibroblasts, it is considered impossible to reflect the function of nerve cells that control the circadian clock. Meanwhile, in 2004, Yoo et al. Created a transgenic mouse (PER2 :: LUC mouse) in which the luciferase gene was knocked in downstream of the mouse Period2 gene, and cultured the collected tissue to monitor the circadian rhythm of luciferase activity. By doing so, it became possible to investigate the expression rhythm of PERIOD2 protein in whole body cells (Non-patent Document 8). It is possible to collect the nerve tissue of the suprachiasmatic nucleus, which is the center of the mammalian biological clock from this mouse, and examine the biological clock function of the nerve cells in the tissue culture system, but in the primary culture system the individual that becomes the donor It is difficult to examine the circadian clock function using nerve cells in a stable state in which reproducibility is ensured depending on the state of the sample and the conditions for collection. When examining the control action of circadian rhythm cycle by harmine, the present inventors can proliferate from PER2 :: LUC mice for the purpose of reproducing the biological clock function of neurons in a stable state in a culture system. We successfully collected and stored neural progenitor cells, induced differentiation into neurons during the experiment, and succeeded in establishing a test system for stably evaluating the biological clock function of neurons (Non-patent Document 9).

Japanese National Patent Publication No. 8-502259 WO2003 / 010168 pamphlet WO2005 / 097101 pamphlet WO2005 / 094849 pamphlet JP 2011-26270 A

"Circadian Rhythm Sleep Clinic" Shigeru Chiba, Kenichi Honma, Emerging Medical Publishers, 2003 Pannain S. et al., The connection between sleep loss, obesity, and type 2 diabetes, P.J. Shiromani et al. (Eds.) Sleep Loss and Obesity: Intersecting Epidemics, Springer Science “Sleep Disorder Response and Treatment Guidelines”, Makoto Uchiyama, Jiho, 2002 Young Ho Rhee, et al., Psychopharmacology, 1990, 101, 486-488 Hidehiko Hibino, Food Style 21, 2003, 7 (3), 50-53 Mariko Izumo, et al., Proc. Natl. Acad. Sci. USA, 2003, 100, 16089-16094 Hiroki R. Ueda, et al., Nature, 2002, 418, 534-539 Seung-Hee Yoo, et al., Proc. Natl. Acad. Sci. USA, 2004, 101, 5339-5346 Daisuke Kondoh, et al., Biol. Pharm. Bull., 2014, 37, 1422-1427

  The object of the present invention is to control the period length or amplitude of circadian rhythms (rhythms controlled by the expression rhythm of a series of clock genes) that are derived from natural ingredients that have been widely consumed so far. An object of the present invention is to provide a circadian rhythm improving agent comprising an active ingredient as an active ingredient, and a food and drink and a pharmaceutical composition containing the same.

  As a result of intensive studies to develop a drug that improves the circadian rhythm, the present inventors are a plant of the family Omodaka, which has a diuretic action, a cholesterol-lowering action, an antihypertensive action, etc., and is a Chinese medicine Hachimi Discovered that herbal medicines such as Jihuangmaru, Gojosan, Tokiyakuyakusan etc. have been traditionally dispensed with, and that allyols contained in Sagomodaka or other genus plants, have their effects, and completed the present invention. It came to do.

That is, the present invention
[1] A circadian rhythm improving agent comprising an alisol or a pharmaceutically acceptable salt thereof.

Here, according to one embodiment of the circadian rhythm improving agent of the present invention,
[2] The circadian rhythm improving agent has an action of regulating the amplitude and / or cycle length of the expression rhythm of a clock gene.
Moreover, according to one embodiment of the circadian rhythm improving agent of the present invention,
[3] The circadian rhythm improving agent according to [2], wherein the clock gene is a Per2 gene.
Moreover, according to one embodiment of the circadian rhythm improving agent of the present invention,
[4] The circadian rhythm improving agent according to any one of [1] to [3], wherein the allisoles are allisole A, allisole B, allisole C, or a derivative thereof.
Moreover, according to one embodiment of the circadian rhythm improving agent of the present invention,
[5] The circadian rhythm improving agent according to any one of [1] to [4] is in the form of a food or drink.

According to another aspect of the present invention, the present invention provides:
[6] A pharmaceutical composition for treating or preventing a circadian rhythm disorder or a disease caused thereby, comprising the circadian rhythm improving agent according to any one of [1] to [4].
According to another aspect of the present invention,
[7] The present invention relates to a food or drink containing the circadian rhythm improving agent according to [1] to [4].
Here, according to one embodiment of the food and drink of the present invention,
[8] The food or drink according to [7] is for a subject having or possibly having a circadian rhythm disorder or a disease caused thereby.
Moreover, according to one embodiment of the food and drink of the present invention,
[9] The food / beverage product according to [7] or [8] is provided with a display indicating that it is used for improving the circadian rhythm.

According to another aspect, the present invention provides:
[10] An agent for improving the expression rhythm of the Per2 gene comprising alisol or a pharmaceutically acceptable salt thereof.
Here, according to the expression rhythm improving agent of Per2 gene of the present invention,
[11] The allysole is alisol A, alisol B, alisol C, or a derivative thereof.

According to another aspect, the present invention provides:
[12] The present invention relates to a circadian rhythm improving method including a step of orally ingesting a circadian rhythm improving agent containing an alisol or a pharmaceutically acceptable salt thereof.

Here, according to one embodiment of the circadian rhythm improvement method of the present invention,
[13] The circadian rhythm improving method according to [12] does not include medical practice.
Moreover, according to one embodiment of the circadian rhythm improving method of the present invention,
[14] The circadian rhythm improving method according to [12] or [13], wherein the step of orally ingesting the circadian rhythm improving agent has or may have a circadian rhythm disorder or a disease caused thereby. The present invention relates to a circadian rhythm improvement method applied to a subject.

  The circadian rhythm-improving agent according to the present invention comprises an allysole or a salt thereof contained in a herbal medicine, its original plant, Sagiomodaka or other related plants, as an active ingredient, so that the amplitude and / or period in the expression rhythm of the clock gene. The length can be adjusted. This can be used to adjust the circadian rhythm decay and period disturbance to normalize the circadian rhythm. The herbal medicine used in the present invention, the original plant Sagimodaka or other allied plants contained in the same genus, have been used as a prescription ingredient for traditional Chinese medicine since ancient times, has high safety, and the circadian rhythm improving agent of the present invention Is useful without problems in use.

FIG. 1 is a graph showing the results of observing the fluorescence intensity of luciferase over time when neural progenitor cells derived from PER2 :: LUC C57BL / 6j mice were cultured in a differentiation medium supplemented with alisol A and luciferin. FIG. 2 is a graph showing the results of observation of luciferase fluorescence intensity over time when neural progenitor cells derived from PER2 :: LUC C57BL / 6j mice were cultured in a differentiation medium supplemented with alisol B and luciferin. FIG. 3 is a graph showing an average value of the cycle length of luciferase expression in each of the group to which Alisol A is added and the vehicle group. FIG. 4 is a graph showing the average value of the cycle length of luciferase expression in each of the group to which Alisol B is added and the vehicle group.

Sleep disorders are roughly classified into “circadian rhythm sleep disorders” and “sleep disordered breathing”. Circadian rhythm sleep disorders are divided into exogenous acute syndromes and intrinsic chronic syndromes, such as sleep phase advance syndrome and sleep phase regression syndrome, non-24-hour sleep-wake disorder, irregular sleep-wake disorder, Included in intrinsic chronic syndrome. The circadian rhythm sleep disorder is thought to involve the body clock. In particular, the intrinsic chronic syndrome is partly due to an abnormality in the body clock. Conventional treatment methods for sleep disorders, such as light irradiation, melatonin administration, and treatments with various sleeping pills are coping therapy that acts on the reset of the body clock, whereas the present invention aims at The sleep disorder treatment method is a treatment method by improving circadian rhythm amplitude abnormality or cycle abnormality, that is, “circadian rhythm disorder”.
Diseases resulting from circadian rhythm disorders are typically considered to be sleep phase advance syndrome (ASPS), sleep phase regression syndrome (DSPS), non-24 hour sleep phase regression syndrome, and seasonal depression. Other periodic and repetitive disorders such as endogenous manic depression, periodic tension, periodic hypertension, periodic ulcer, irregular ovulation cycle, and diabetes caused by periodic abnormalities of insulin secretion, and cerebral vascular types Circadian rhythm disorder is also considered to be involved in dementia and nighttime hemorrhage in Alzheimer type dementia. In addition, attenuation of amplitude and cycle disturbance of various circadian rhythms are related to metabolic diseases such as aging, diabetes and obesity. The present invention increases the circadian rhythm amplitude (amplitude of clock gene expression rhythm) among circadian rhythm disorders, or shortens the circadian rhythm cycle length (cycle length of clock gene expression rhythm). It is intended to improve obstacles.

The circadian rhythm improving agent of the present invention regulates the amplitude and / or cycle length of the expression rhythm of the clock gene. The circadian rhythm improving agent of the present invention preferably regulates both the amplitude and cycle length of the expression rhythm of the clock gene, but can regulate either the amplitude or cycle length of the expression rhythm of the clock gene. Are also included in the circadian rhythm improving agent of the present invention.
In the present specification, “to regulate the amplitude of the expression rhythm of the clock gene” means that when the clock gene shows activation and suppression of expression along a certain expression rhythm (cycle), It means that the expression is further activated or further suppressed. Therefore, a preferred embodiment of “adjusting the amplitude of the expression rhythm of the clock gene” is to improve the attenuation of the amplitude of the expression rhythm (increase the amplitude). Circadian rhythm improving agent of the present invention, in particular, the expression of Per2 gene of clock genes, (increasing the expression amount) further activates along the expression rhythm of the gene can be and this.
Further, `` regulate the cycle length of the expression rhythm of the clock gene '' means that when the clock gene shows activation of expression and suppression of expression along a certain expression rhythm (cycle), the length of the expression rhythm ( It means to extend or shorten the (cycle length). The circadian rhythm improving agent of the present invention can particularly shorten the length (cycle length) of the expression rhythm of Per2 in the clock gene.

The clock gene is a gene that controls biological rhythm, especially circadian rhythm (circadian rhythm), and is also called circadian rhythm control gene. Clock genes include Cry gene (Cry, Cryptochrome) (Cry1, Cry2, etc.), Clock gene (CLOCK), Dec gene (Dec), Rev-erb gene (Rev-erb) (Rev-Erbα gene, etc.), CK1 gene (CK1, Casein Kinase 1) [CK1ε, CK1δ etc.], Bmal gene (Bmal) [Bmal1 etc.], Per gene (Per, Period) [Per1, Per2 etc.]. These genes are known to be possessed by various animals such as humans and mice.
The circadian rhythm is composed of the negative feedback loop mechanism between the above clock genes (Molecular components of the Mammalian circadian clock. Buhr ED, Takahashi JS. Handb Exp Pharmacol. 2013; (217): 3-27). By controlling the expression of the Per2 gene, which plays a role, the expression of a series of clock genes is controlled, and as a result, the circadian rhythm can be controlled.

  The circadian rhythm improving agent of the present invention contains alisol or a pharmaceutically acceptable salt thereof. The compound of alisols is a compound contained in Sagiomodaka, which is the original plant of herbal medicine, or other related plants. Conventionally, allisols (eg, allisole A, allisole B, and allisole C, and derivatives thereof) contained in Sagiomodaka or other related plants have been used as prescription ingredients for traditional Chinese medicine since ancient times. Is expensive. As shown in usage examples in Kampo, compounds contained in alisols are used as having similar actions because they have similar skeleton parts. For example, Japanese Patent Application Laid-Open No. 08-259455 discloses that alisols (alisol A, alisol B, and their monoacetate derivatives) can be extracted from a lot and used as an agent for improving nephritis. In Kaihei 07-173188, a series of compounds of arisols isolated from potato (for example, alisol B, alisol B monoacetate, 11-deoxy alisol B, 16-hydroxy alisol B monoacetate, alisol C mono Acetate) is used as a brain function improving agent.

In the present specification, “alisols” include compounds such as allisole A, allisole B, allisole C, allisole D, and derivatives and stereoisomers thereof. “Derivatives or stereoisomers of compounds such as alisol A, alisol B, alisol C, and alisol D” include those in which the hydroxyl group is substituted with a hydrogen atom (deoxy derivative) in the structural formula of the compound, and the hydroxyl group is carboxyl Examples thereof include those substituted on a group (acetate derivatives) and epimerized ones.
Examples of the alisols used in the circadian rhythm improving agent of the present invention include alisol A or alisol B represented by the following formula I or II.

式II(アリソールＢ)：

Formula I (Arisol A):

Formula II (Arisol B):

  In addition to allisole A and allisole B, allisols used in the circadian rhythm improving agent of the present invention include allisol as long as it has a circadian rhythm improving action (or a clock gene expression rhythm regulating action). A compound known as A, alisol B, alisol C, alisol D, etc., and derivatives or stereoisomers thereof can be used. Specifically, although not limited to the following, for example, allisol A to P, allisol A23-acetate, allisol A24-acetate, allisol A triacetate, dihydroallylsol A, epialisole A, allisol B acetate, 11-deoxyallylsol B 11-deoxy allysole B23-acetate, 16β-hydroxy allysole B23-monoacetate, allysole C23-acetate, and the like. The allysole used in the circadian rhythm improving agent of the present invention is preferably alisol A, alisol B, alisol C, or a derivative thereof.

  As the compounds of alisols used for the circadian rhythm improving agent of the present invention, commercially available ones may be used, or those isolated and purified from Sadiomodaka or other related plants can also be used. Those skilled in the art can appropriately obtain alisol compounds from scorpionfish or other related plants by known methods. For example, a tuber of Sagiomodaka or other related genera is extracted with a solvent such as hexane, diethyl ether, petroleum ether, ethyl acetate, chloroform, acetone, methanol, ethanol, water, and the residue obtained by removing the solvent from the extract. Thus, the target compound can be obtained by using column chromatography or high performance liquid chromatography (for example, the method described in paragraphs 0017 and 0021 to 0036 of JP-A-07-173188).

  In addition, pharmacologically acceptable salts are not limited to the following, but include, for example, inorganic acid salts such as hydrochloride, sulfate, phosphate, acetate, trifluoroacetate, citrate, maleate And organic acid salts such as fumarate, lactate and tartrate.

  Components other than allisols and pharmacologically acceptable salts thereof (pharmacologically acceptable base) that can be included in the circadian rhythm improving agent of the present invention, unless the object of the present invention is impaired. There is no particular limitation. For example, excipients, disintegrants, binders, lubricants, coating agents, colorants, color formers, flavoring agents, flavoring agents, antioxidants, preservatives, flavoring agents, sour agents, sweeteners, strengthening From various preparations, vitamins, swelling agents, thickeners, surfactants, etc. that do not impair various properties required for the formulation (e.g. storage stabilizers) 1 type or 2 types or more can be selected according to the dosage form of quasi-drugs and the like. In addition, components other than alisols or pharmacologically acceptable salts thereof may be other components having an effect of improving circadian rhythm.

  The circadian rhythm improving agent of the present invention can be used as a final product as it is. The circadian rhythm improving agent of the present invention can be used as an additive for food and drink, a rhythm additive for pharmaceuticals, and an additive for quasi drugs. Thereby, the circadian rhythm improvement effect can be provided as food / beverage products and a pharmaceutical.

  The administration form of the circadian rhythm improving agent of the present invention is not particularly limited. For example, oral administration (eg, oral administration, sublingual administration, etc.), parenteral administration (intravenous administration, intramuscular administration, subcutaneous administration, transdermal administration, nasal administration, pulmonary administration, etc.) and the like can be mentioned. Among these, a less invasive dosage form is preferable, and oral administration is more preferable. When the circadian rhythm improving agent of the present invention is used as a food or drink, it is more preferably administered orally.

  The dosage form of the circadian rhythm improving agent of the present invention can be appropriately determined depending on whether it is used as a food / beverage product, a medicine or a quasi-drug, and is not particularly limited. Examples of dosage forms for oral administration include liquid (liquid), syrup (syrup), tablet (tablet), capsule (capsule), powder (granule, fine granule), soft capsule (soft capsule) Agent), solid, semi-liquid, cream, and paste. Of these, granular, capsule, powder, soft capsule, and solid are preferable, and granular (tablet) is more preferable.

  For example, in order to exert the desired effect as an oral preparation, it varies depending on the patient's age, body weight, and degree of disease, but usually 50 mg to 5 g of the compound of formula is divided into several times a day in adults. It seems appropriate to take. The oral preparation is produced according to a conventional method using, for example, starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, inorganic salts and the like. In this type of preparation, a binder, a disintegrant, a surfactant, a lubricant, a fluidity promoter, a corrigent, a colorant, a fragrance and the like can be appropriately used in addition to the above-mentioned excipients. Specific examples of each are as follows. [Binder] Starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol. [Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethylcellulose, carboxymethylcellulose calcium, carboxymethylcellulose, low-substituted hydroxypropylcellulose. [Surfactant] Sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 80. [Lubricant] Talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol. [Flowability promoter] Light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The circadian rhythm improving agent of the present invention can also be administered as a suspension, emulsion, syrup, or elixir, and these various dosage forms contain a flavoring agent and a coloring agent. Also good. In order to exert the desired effect as a parenteral agent, it varies depending on the patient's age, body weight, and degree of illness, but it is usually an intravenous dose of 0.1 mg to 1 g / day as a weight of the compound of the formula in an adult, intravenous drip infusion Note, subcutaneous injection and intramuscular injection may be appropriate. This parenteral preparation is produced according to a conventional method, and generally used as a diluent is distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol and the like. be able to. Furthermore, you may add a bactericidal agent, antiseptic | preservative, a stabilizer, and an emulsifier as needed. In addition, from the viewpoint of stability, this parenteral preparation can be frozen after filling into a vial or the like, water can be removed by a normal freeze-drying technique, and a liquid preparation can be re-prepared from the freeze-dried product immediately before use. Furthermore, tonicity agents, stabilizers, preservatives, soothing agents and the like may be added as necessary. Examples of other parenteral preparations include coating solutions for external use, ointments and the like, suppositories for rectal administration, and the like, and are produced according to a conventional method.
In addition, the administration time of the circadian rhythm improving agent of the present invention is not particularly limited.

  Moreover, in one embodiment of this invention, the pharmaceutical composition for treating or preventing the circadian rhythm disorder or the disease resulting therefrom including a circadian rhythm improving agent is provided. The said pharmaceutical composition can contain an excipient | filler etc. similarly to the circadian rhythm improving agent, and can also contain the other component which has a circadian rhythm improving effect. In addition, the pharmaceutical composition can be appropriately designed and selected in the same manner as the circadian rhythm improving agent.

According to the circadian rhythm improving agent or pharmaceutical composition of the present invention, it is possible to regulate the expression of a clock gene, and further to regulate the circadian rhythm. Therefore, according to the said rhythm improving agent or pharmaceutical composition, a circadian rhythm disorder or the disease resulting from it can be prevented, relieved, or treated. The disease caused by circadian rhythm disorder refers to a disease caused by disturbance or attenuation of the circadian rhythm cycle, but is not limited to the following, for example, sleep phase regression syndrome (DSPS), sleep phase advance syndrome ( ASPS), non-24-hour sleep-wake disorder, and irregular sleep-wake patterns, sleep disorders and insomnia, autonomic dystonia, manic depression, seasonal depression, periodic tension, periodic hypertension Diseases, periodic ulcers, irregular ovulation cycles, periodic / repetitive disorders such as diabetes due to periodic abnormalities of insulin secretion, nocturnal epilepsy in cerebrovascular dementia and Alzheimer dementia.
Moreover, as one embodiment of the present invention, there is provided an agent for improving the expression cycle of Per2 gene, which comprises alisols or a pharmaceutically acceptable salt thereof. The Per2 gene expression cycle improving agent can be used in the same manner as the above circadian rhythm improving agent.

Moreover, according to one Embodiment of this invention, the food / beverage products containing a circadian rhythm improving agent are provided. The food or drink can be provided as a food or drink for improving the circadian rhythm, such as general food, functional food, health food, health supplement, nutritional supplement, insurance functional food, food for specified insurance, or It can be used as various foods and beverages such as nutritional functional foods (note that in this specification, the term “food and beverage products” does not include medicines including Chinese medicine). Examples of the form of food and drink include beverages (soft drinks, carbonated drinks, nutritional drinks, powdered drinks, fruit drinks, milk drinks, jelly drinks, etc.), confectionery (cookies, cakes, gums, candy, tablets, gummi, buns, etc. , Yokan, pudding, jelly, ice cream, sherbet etc.) Cheese, margarine, fermented milk, etc.), soup (powdered soup, liquid soup, etc.), staple foods (rice, noodles (dried noodles, raw noodles), bread, cereal, etc.), seasonings (mayonnaise, shortening, dressing, sauce) , Sauce, soy sauce, etc.) and supplements.
The food and drink of the present invention can be used for subjects (mammals including humans) having or possibly having the circadian rhythm disorders listed above or diseases resulting therefrom.

  In the present invention, the circadian rhythm improving action is an action of shortening the period length of the circadian rhythm controlled by the expression of a clock gene such as Per2 and / or an action of enhancing the amplitude of the period. In order to confirm these effects, for example, as in the method disclosed in Non-Patent Document 9, the influence of a candidate substance on the expression cycle of a reporter gene (such as a luciferase gene) whose expression is controlled by a Per2 promoter in a cell. It can be confirmed by examining.

  The terms used in the present specification are used to describe specific embodiments, and are not used with the intention of limiting the present invention.

  In addition, the term “comprising” in the present specification intends that there are items (steps, elements, etc.) described unless there is a case where interpretation should be clearly different from the description in this specification, and It does not exclude the presence of other matters (processes, elements, etc.).

  Moreover, unless otherwise defined, terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms used herein should be interpreted as having a meaning consistent with the meaning in this specification and the related technical field, unless otherwise defined, idealized, or overly formal. It should not be interpreted in a general sense.

  In the following, the present invention will be described in more detail with reference to examples. However, the invention can be implemented in various ways and should not be construed as limited to the embodiments set forth herein.

(Example 1) Periodic rhythm reduction and amplitude enhancement of circadian rhythm by alisols Embryonic 14-day-old PER2 :: LUC C57BL / 6J mice (obtained from Jackson Laboratories; see Non-patent Document 8) were taken out and 0.05% The mixture was heated at 37 ° C. for 10 minutes with a PBS solution containing trypsin (Sigma) and DNase I (1 mg / L, Gibco). Disperse the tissue by pipetting, and after centrifugation, the neural stem cells can be transformed into a stem cell medium (20ng / ml basic fibroblast growth factor (bFGF) (R & D Systems), 20ng / ml epidermal growth factor (EGF) (R & D Systems), B27 supplements The suspension was suspended in a DMEM / F12 medium (Gibco) containing (Gibco)] and seeded on a plate coated with poly-HEMA (Sigma) at a concentration of 5 × 10 ⁵ cells / ml. Neural progenitor cells were formed by the formation of neural stem cell clusters and stored frozen until use.

Neural progenitor cells used for the experiments were those thawed from a cryopreserved state. The neural progenitor cells after thawing were cultured in a stem cell medium for 24 hours, and then induced to differentiate for 7 days in a differentiation medium [DMEM / F12 medium containing 2% fetal calf serum and B27 supplements].
A candidate substance was searched using differentiated nerve cells. Differentiation of neurons with differentiation medium containing HEPES (10 mM) and luminescent substrate luciferin (0.1 mM, Wako Pure Chemicals), plus candidate substances along with forskolin (10 μM, Sigma) to synchronize the body clock The luminescence was measured with luciferase over time using LM2400 (Hamamatsu Photonics).
As candidate substances, alisol A and alisol B (purchased from Wako Pure Chemical Industries, Ltd.) were used. Allisole A or allisole B was added alone to the differentiation medium to a final concentration of 10 μM. It has been found that when neurons are cultured using a differentiation medium supplemented with alisol A or alisol B, the cycle of the luminescence rhythm is shortened. The results are shown in FIG. 1 (result when adding allisole A) and FIG. 2 (result when adding allisole B). 3 and 4 show the average value of the expression cycle of luciferase automatically calculated by LM2400 (Hamamatsu Photonics), respectively, in the group to which allisole A or allisole B is added. As shown in FIG. 3 and FIG. 4, in the group to which Allisol A was added and in the group to which Allisol B was added, a shortening of the cycle length of about 1 hour was observed as compared with the Vehicle group.
In addition, the amplitude increase effect of the light emission rhythm was also confirmed in FIGS.

Claims (2)

A circadian rhythm modulator comprising alisol A or alisol B or a pharmaceutically acceptable salt thereof , wherein the regulation is an increase in the amplitude of the expression rhythm of the Per2 gene and / or a shortening of the cycle length. Rhythm regulator . A Per2 gene expression rhythm regulator comprising Arisol A or Arisol B or a pharmaceutically acceptable salt thereof , wherein the regulation is an effect of increasing the amplitude of the expression rhythm of the Per2 gene and / or shortening the cycle length. Expression rhythm regulator .

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