JP6049289B2 - Circadian rhythm regulator - Google Patents

Description

  The present invention relates to a circadian rhythm regulator by regulating the expression level of a mammalian clock gene. More specifically, the present invention relates to a circadian rhythm regulator by regulating the expression level of clock genes Per and Bmal.

Many biological activities show autonomous periodic fluctuations called biological rhythms. Among them, a rhythm that shows a period of about 24 hours is called a circadian rhythm.
In addition to individual behavior and activity, various physiological phenomena such as body temperature, hormone secretion, and metabolism show circadian rhythms. In modern life, there are many problems caused by disturbance of circadian rhythm such as sleep disturbance caused by irregular life patterns such as shift work, jet lag caused by airplane movement. Moreover, in elderly people, a decrease in the synthesis of endogenous factors (eg, melatonin) for forming circadian rhythms is observed, and there is concern about a decrease in QOL due to a decrease in circadian rhythm maintenance ability (non-patent literature). 1, Non-Patent Document 2). Therefore, normal adjustment of the circadian rhythm is an important issue.

  As a method for adjusting the circadian rhythm, administration of melatonin or melatonin derivatives which are endogenous hormones has been attempted so far. However, since administration of hormones also affects various physiological phenomena other than circadian rhythm, problems of side effects have been pointed out (Non-patent Documents 3 and 4).

  In recent years, it has become clear that the circadian rhythm is regulated by a group of genes called clock genes. As specific clock genes, a Per gene (Per1, Per2, Per3), a Clock gene, a Bmal gene (Bmal1, Bmal2, Bmal3), a Cry gene (Cry1, Cry2) and the like have been reported. Among them, it has been found that the Per gene and the Bmal gene are important clock genes. In mammals, the Per gene and the Bmal gene are expressed with a circadian rhythm at a phase shifted by approximately 12 hours, and Bmal expression increases in the middle of the night, while Per gene expression increases in the daytime (Patent Document 1). . Deviation in the phase of the expression rhythm of the Per gene and the Bmal gene leads to disturbance of circadian rhythm. Therefore, if the expression rhythm of these genes can be synchronized with the normal phase by adjusting the expression level of the Per gene and Bmal gene, it is considered that the circadian rhythm cycle can be effectively adjusted. . A circadian rhythm regulator that can effectively synchronize the phase of the expression rhythm of the Per gene and the Bmal gene with a normal state, is highly safe, and can be taken daily is desired.

WO2011 / 122041

Vitiello MV., Clin Corne rstone 2000; 2 (5): 16-27 Mishima K. et al., Chronobiol Int 2000 May; 17 (3): 419-32 Monti JM, Cardinali DP., Biol Signals Recept 2000, 9: 328-39 Reiter RJ., Ann Med 1998, 30: 103-8

  An object of the present invention is to provide a circadian rhythm regulator that is highly safe, can be ingested on a daily basis, and is highly effective by regulating the expression of clock genes, particularly the Per gene and Bmal gene. And

  As a result of searching for effective ingredients from natural food materials with abundant food experience and high safety, the present inventors have found that oleacein, which is a polyphenol extracted from olives, has clock genes Per and Bmal. Was found to have an effect of regulating the expression level of. Oreacein can synchronize the expression rhythms of the Per and Bmal genes to normal phases by adjusting the expression levels of the two genes Per and Bmal in the opposite phase, and adjust the circadian rhythm normally. Can do.

That is, the present invention includes, but is not limited to, the following.
[1] A circadian rhythm regulator containing oleacein.
[2] The circadian rhythm adjusting agent according to [1], wherein the expression levels of the Per1 gene and the Bmal1 gene are adjusted to an opposite phase, thereby synchronizing the expression rhythm of the Per gene and the Bmal gene with a normal phase.
[3] A circadian rhythm adjusting agent containing an olive extract.
[4] The circadian rhythm adjusting agent according to [3], wherein the olive extract contains 30% by weight or more of olive-derived polyphenol.
[5] The circadian rhythm adjusting agent according to [4], wherein the olive-derived polyphenol contains oleacein.
[6] The circadian rhythm adjusting agent according to any one of [3] to [5], wherein the olive extract is obtained by extracting olive fruit with water, ethanol, or a mixture thereof.
[7] The olive extract is a 40% to 50% ethanol extract fraction of a defatted residue of olive fruit extracted with hot water at 70 ° C to 100 ° C, according to any one of [3] to [5] Circadian rhythm regulator.
[8] A food or drink to which the circadian rhythm adjusting agent according to any one of [1] to [7] is added.
[9] A pharmaceutical composition for prevention, alleviation, or treatment of a physical condition or disease caused by disturbance of circadian rhythm, comprising oleacein.
[10] A method for producing a food or drink or pharmaceutical composition having a circadian rhythm adjusting action, comprising a step of blending oleacein.
[11] Use of oleacein in the production of a food or drink or a pharmaceutical composition having a circadian rhythm adjustment effect.

  The circadian rhythm adjusting agent of the present invention can regulate the expression levels of two genes, Per and Bmal, in reverse phase by including oleacein, whereby the expression rhythm of Per gene and Bmal gene can be adjusted to normal phase. The circadian rhythm can be adjusted effectively. Moreover, since the circadian rhythm regulator of this invention uses the oleacein contained in olive as an active ingredient, it has abundant eating experience, there are few side effects, and its safety is high.

FIG. 1 shows the gene expression level of the clock gene Bmal1 in HepG2 cells supplemented with various olive-derived polyphenols such as oleacein. FIG. 2 shows the gene expression level of the clock gene Per1 in HepG2 cells to which various olive-derived polyphenols such as oleacein were added. FIG. 3 shows the gene expression level of the clock gene Bmal1 in the liver of a Wistar rat administered with an oleacein-rich olive extract. FIG. 4 shows the gene expression level of the clock gene Per1 in the liver of a Wistar rat administered with an oleacein-rich olive extract.

<Circadian rhythm regulator>
The circadian rhythm adjusting agent of the present invention can control the expression level of the Per gene and Bmal gene, which are clock genes, in the opposite phase by including oleacein as an active ingredient, thereby The circadian rhythm can be adjusted normally by synchronizing the expression rhythm to the normal phase. Therefore, the circadian rhythm adjusting agent of the present invention can be effectively used for the prevention, alleviation, treatment, etc. of physical disorder and diseases caused by disturbance of circadian rhythm such as sleep disorder and jet lag. Can do.
<Per gene and Bmal gene>
The circadian rhythm adjusting agent of the present invention can regulate the expression of the clock gene by including oleacein as an active ingredient, and in particular, can regulate the expression level of the Per gene and the Bmal gene in the opposite phase.

  Clock genes are a group of genes that act to regulate circadian rhythm. When these genes have mutations, they are defined as genes that affect behavioral rhythms (Ishida, 2009, Biochemistry, Vol. 81). , No. 2, pp. 75-83). Among the major factors are the Period (Per) gene and the Bmal gene.

  In mammals, Period1 (per1), period2 (per2), and period3 (per3), which are Period gene families, are known to function as clock genes. In addition, Bmal1, a Bmal gene family, is known to function as a clock gene. BMAL1, a Bmal1 gene product, forms a heterodimer with CLOCK to promote Period gene transcription, while PERIOD, a Period gene product, forms a heterodimer with CRYPTOCHROME to activate BMAL1 and CLOCK Suppress.

In mammals, the Per gene and the Bmal gene are expressed in a circadian rhythm with phases shifted by approximately 12 hours, respectively, and Bmal expression increases in the night, while Per gene expression increases in the daytime. This is the normal phase of the expression rhythm of the Per gene and Bmal gene. Deviation in the phase of the expression rhythm of the Per gene and the Bmal gene leads to disturbance of the circadian rhythm. The circadian rhythm adjusting agent of the present invention can synchronize the expression rhythms of the Per gene and the Bmal gene to the normal phase by adjusting the expression levels of the Per gene and the Bmal gene in the opposite phase.
<Performance analysis of Per gene and Bmal gene>
The expression levels of the Per gene and Bmal gene regulated by the circadian rhythm regulator of the present invention can be analyzed using any method known to those skilled in the art. For example, methods such as Northern blotting analysis using an arbitrary sequence in the Per or Bmal gene sequence as a probe, real-time RT-PCR analysis, and expression level analysis using a DNA microarray can be suitably used. Any sample can be used as a sample for gene expression analysis as long as it contains cells. For example, hair containing hair follicle cells, biopsy such as skin, blood, saliva, etc. can be used. .
<Oreacein>
Oreacein (CAS number: 149183-75-5) is one of the polyphenol components abundantly contained in olives and has the following structural formula.

  In the present invention, not only oleacein but also a modified oleacein derivative or an arbitrary salt form can be used. The oleacein derivative includes a group of compounds obtained by chemically modifying the hydroxyl group of oleacein and includes oleacein esters and glycosides.

  In the circadian rhythm adjusting agent of the present invention, plant materials such as olives rich in oleacein as an active ingredient may be used as they are, or high oleacein content extracted from plant materials such as olives may be used.

  When a plant material rich in oleacein is used as it is as a circadian rhythm regulator of the present invention, for example, olive fruits, seeds, leaves, stems, etc. are used raw or dried by freeze drying, etc. Can do. When olive leaves are used, they may be mixed with leaves of other plant species, such as tea leaves. Also, olive oil extracted from olive fruit can be used as a plant raw material rich in oleacein. As the olive oil, a commercially available product may be used, or it may be prepared from olives by a known method. In addition, an oleacein-rich olive extract can be prepared from plant materials such as olives by the method described later.

The content of oleacein, which is an active ingredient of the circadian rhythm adjusting agent of the present invention, can be arbitrarily determined in consideration of the effect, but 0.01 to 40% by weight is contained in the circadian rhythm adjusting agent. Is preferably included, more preferably 0.1 to 30% by weight, and even more preferably 1 to 20% by weight.
<Olive>
As the circadian rhythm adjusting agent of the present invention, olive fruits, leaves, seeds, leaves, stems and the like as plant materials rich in oleacein may be used as they are, or oleacein high-content olive extract may be used. . Any type of olive can be used, but for example, varieties such as Manzanillo, Lucca, Nevadillo Blanco, Mission, Picual, Arbequina, Ohiblanca, Cornicabra, Gordal, Moloiolo, Frantoio, Colatina, Retino, etc. Can do.

For preparing the oleacein-rich olive extract, any part of olive fruit, seeds, leaves, stems, etc. may be used, but it is preferable to use fruits. Olive fruit may be used as it is, or may be dried by freeze-drying or the like. Moreover, the residue after squeezing oil from olive fruit can be used as it is or in a dried state.
<Oreacein high content olive extract>
For the preparation of the oleacein-rich olive extract, any method may be used as long as an extract rich in olive-derived polyphenols can be obtained. The olive-derived polyphenol referred to here is a polyphenol contained in olive, which includes a hydroxytyrosol skeleton as a part of the structure, a tyrosol skeleton as a part of the structure, and a caffeic acid skeleton as a part of the structure. Including a p-coumaric acid skeleton as a part of the structure, or a mixture thereof. Examples of olive-derived polyphenols that contain a hydroxytyrosol skeleton as part of the structure include hydroxytyrosol and its precursors and metabolites. Examples of olive-derived polyphenols that contain a tyrosol skeleton as part of the structure include tyrosol and its precursors and metabolites. Examples of olive-derived polyphenols that include a caffeic acid skeleton as part of the structure include caffeic acid and its precursors and metabolites. Examples of olive-derived polyphenols containing a p-coumaric acid skeleton as part of the structure include p-coumaric acid and its precursors and metabolites. The olive-derived polyphenol preferably contains oleacein in an amount of 0.01 to 40% by weight, more preferably 0.1 to 30% by weight, and still more preferably 1 to 20% by weight.

The oleacein high-content olive extract of the present invention can be prepared by a method that provides an extract rich in olive-derived polyphenols, preferably 30% by weight or more. Specifically, for example, an oleacein-rich olive extract can be prepared by the following method.
(1) The residue after squeezing olive oil from olive is defatted using an organic solvent, and then extracted using hot water. Any organic solvent may be used for degreasing, but for example, hexane can be used. The temperature of hot water used for extraction is preferably 70 ° C to 100 ° C, and most preferably 70 ° C to 80 ° C.
(2) Extract containing a large amount of olive-derived polyphenols by loading the extract obtained in step (1) onto a column packed with adsorption resin and eluting with water, ethanol, or a mixture thereof Get the fraction. Since the extract fraction rich in olive-derived polyphenol also contains abundant oleacein, this extract fraction can be used as the circadian rhythm regulator of the present invention as an oleacein-rich olive extract. Any adsorbent resin may be used as long as it adsorbs polyphenol. For example, Amberlite (trademark) XAD-HP (Oregano) can be suitably used. Moreover, about the elution using water, ethanol, or these mixtures, those skilled in the art can set the conditions suitably according to the variety, form, etc. of the olive to be used. When using the residue after squeezing olive oil for extraction, for example, stepwise elution of water, 15% ethanol / water, 30% ethanol / water, 45% ethanol / water, and the 45% ethanol fraction obtained The fraction can be used in the present invention as an oleacein rich olive extract.

Moreover, as an oleacein high-content olive extract, the product marketed as an Orea Select (trademark) from Indena can also be used suitably.
The concentration of oleacein contained in the oleacein high-content olive extract is preferably 0.01 to 40% by weight, more preferably 0.1 to 30% by weight, and 1 to 20% by weight. Further preferred.
<Food and drink with circadian rhythm modifier added>
The circadian rhythm adjusting agent of the present invention can be added to food and drink. That is, this invention provides the food / beverage products which added the circadian rhythm regulator. For example, supplements containing oleacein as an active ingredient, as well as functional foods that contain oleacein in general foods and drinks, and that provide food and drinks with a circadian rhythm adjustment effect (health supplements, nutritional functional foods, special-purpose foods, specific foods) Health foods such as health foods, including animal supplements), animal feeds, and the like can be used as ingested forms. The food / beverage products to which the circadian rhythm adjusting agent of the present invention is added can display the kind of active ingredient, the use, the effect on circadian rhythm adjustment, and / or the ingestion method on the packaging, container or instructions.

  The food and drink to which the circadian rhythm adjusting agent of the present invention is added may be in any form, for example, beverages such as juice, milk, coffee beverages, tea beverages, liquid foods such as soups, and pasty foods such as yogurt. , Semi-solid foods such as jelly and gummi, solid foods such as cookies and gums, and fat-containing foods such as dressings and mayonnaise. Further, oleacein can be added to tea leaves or the like mixed with olive leaves to make a food or drink with enhanced circadian rhythm effect.

  The food and drink to which the circadian rhythm adjusting agent of the present invention is added do not impair the effect of oleacein, that is, as long as it does not cause an unfavorable interaction by blending with oleacein, a mineral; vitamin E, vitamin Vitamins such as C and vitamin A; nutritional ingredients; fragrances; other additives such as pigments can be mixed. Any of these additives commonly used for food and drink can be used. Moreover, as long as the effect of oleacein is not impaired, other physiologically active ingredients can be mixed in the food and drink to which the circadian rhythm adjusting agent of the present invention is added.

The food or drink to which the circadian rhythm modifier of the present invention is added is preferably oleacein, preferably 0.001 to 10% by weight, more preferably 0.01 to 10% by weight, still more preferably 0.1 to 5% by weight. contains.
<Oleacein-containing pharmaceutical composition for preventing, alleviating, or treating disorders of physical condition or diseases caused by disturbance of circadian rhythm>
The pharmaceutical composition of the present invention contains oleacein as an active ingredient, so that the disorder of physical condition caused by disturbance of circadian rhythm such as sleep disorder caused by irregular life patterns and jet lag caused by airplane movement. Or it can be used to prevent, alleviate or treat the disease.

  The pharmaceutical composition of the present invention can be formulated according to the purpose by a general method together with a pharmacologically acceptable carrier, diluent or excipient. Examples of diluents and carriers include liquid diluents such as water, ethanol, propylene glycol and glycerin, solid diluents and excipients such as glucose, sucrose, dextrin, cyclodextrin and gum arabic. In addition, emulsifiers, tonicity agents (isotonic agents), buffers, solubilizers, preservatives, stabilizers, antioxidants, and the like that are generally used in formulation can be appropriately blended.

  The pharmaceutical composition of the present invention does not impair the effects of oleacein, that is, if necessary, minerals; vitamins such as vitamin E, vitamin C, vitamin A, etc. Nutrients; flavors; other additives such as pigments can be mixed. Any of these additives commonly used in pharmaceuticals can be used. In addition, other physiologically active ingredients can be mixed into the pharmaceutical composition of the present invention as long as the effect of oleacein is not impaired.

  The form of the pharmaceutical composition of the present invention is not particularly limited, and is, for example, a solid form such as a powder, a granule, or a tablet; a liquid such as a solution, an emulsion, or a dispersion; or a paste It can be prepared in any form such as a semi-solid form. Specific examples of the dosage form include powders, granules, fine granules, tablets, pills, troches, capsules (including soft capsules and hard capsules), chewable agents, and solutions.

The pharmaceutical composition of the present invention contains oleacein preferably at 0.1 to 50% by weight, more preferably 1 to 10% by weight.
What is necessary is just to select suitably the dosage amount and dosage form of the pharmaceutical composition of this invention according to a subject, a disease state, its progress, and other conditions. For example, when orally administered to humans (adults) for the purpose of obtaining a circadian rhythm adjustment effect, generally oleacein is 0.01 to 40 mg per day, preferably 0.1 to 30 mg, more preferably 1 to It is good to administer continuously about 1 to 3 times a day so that it may become about 20 mg.
<Manufacture of food and drink or pharmaceutical composition having a circadian rhythm adjustment effect>
The circadian rhythm adjusting agent of the present invention can regulate the expression levels of the Per gene and Bmal gene, which are clock genes, in reverse phase by including oleacein as an active ingredient. And it can provide as food-drinks or a pharmaceutical composition.
That is, this invention is a manufacturing method of the food-drinks or pharmaceutical composition which has a circadian rhythm adjustment effect | action including the process of mix | blending oleacein from another viewpoint. Furthermore, it is use of oleacein in manufacture of the food-drinks or pharmaceutical composition which has a circadian rhythm adjustment effect | action from another viewpoint.

  Hereinafter, the present invention will be described more specifically based on examples. The present invention is not limited to these examples.

Test Example 1 Circadian rhythm control effect of olive-derived components
The circadian rhythm adjustment effect of the olive-derived component was evaluated by the following method using the expression of the clock gene as an index.
<Test substance>
Commercially available luteolin (Funakoshi), apigenin (Funakoshi) and caffeic acid (Nacalai Tesque) were used. Oreacein and acteoside were prepared as follows.
<Preparation method>
100 g of olive extract extract (Orea Select (Batch No. 10219) manufactured by Indena) is dissolved in 1200 ml of methyl ethyl ketone and 600 ml of distilled water, and shaken well with a 3 L separatory funnel. The layer was collected. To the aqueous layer, 1200 ml of methyl ethyl ketone was added and shaken well to recover the organic layer, and the same operation was repeated. A total of 3 organic layers were combined, concentrated and freeze-dried to obtain 52.5 g of a dried product.

  The total amount of the dried product was dissolved in 200 ml of 70% ethanol aqueous solution, further diluted 10-fold with water, and then equilibrated with water. 1 L of MCI GEL CHP-20P (Mitsubishi Chemical Corporation, 75-150 μm ). After flowing 10 L of 10% ethanol aqueous solution, 1 L each in 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 35% ethanol aqueous solution, 2 more Elution was sequentially performed with 40% ethanol aqueous solution of L (all ethanol concentrations were V / V%). The eluate from 12.5% to 27.5% was collected in 4 fractions of 250 ml each. Of these, the 4th fraction of 20% eluate to the 3rd fraction of 25% eluate were combined and concentrated to 5.98 g Purified acteoside was obtained.

From the 4th fraction of 25% eluate to the 2nd fraction of 30% eluate were combined and concentrated to obtain a fraction containing 5.56 g of oleacein. This fraction was purified by preparative HPLC under the following conditions.
<Conditions>
Column: Develosil ODS-HG-5 (5cmφx50cm, Nomura Chemical Co., Ltd.)
Mobile phase: A: 0.05% TFA / H2O, B: 90% CH3CN, 0.05% TFA / H2O, 30ml / min
Gradient program: B20% → B45% (90min), B45% isocratic (150min)
Detection: A280nm
Under this HPLC condition, oleacein eluted from 60 to 70 minutes was collected, concentrated under reduced pressure, and lyophilized to obtain 3.8 g of oleacein.
<Evaluation method>
Cultured cells (human liver cancer-derived cell line HepG2) were seeded in T-75 flasks (BD Falcon (trademark)) at 2.5 × 10 3 cells / cm ² and medium exchange (RPMI1640, 10% FBS) was performed every 3 days. Passage 80% confluent cells to a 6 well plate (BD Falcon ™) at a concentration of 4.2x10 ⁴ cells / ml and add the medium after 3 days to synchronize the circadian rhythm phase of each cell. The medium was replaced with low serum medium (RPMI1640, 2% FBS). Cells were subjected to experiments 3 days after low serum medium replacement. A DMSO solution (50 μg / μl) of the test substance was prepared, and this solution was added to a 6-well plate so that the final concentration of the test substance was 50 μg / ml. DMSO was added to the control (final concentration 0.1%).

Cells were collected 8 hours after addition of the test substance, and mRNA was extracted according to a conventional method. Using the extracted mRNA, the expression levels of clock genes Bmal1 and Per1 were analyzed by qPCR (Applied Biosystems 7900HT Fast real-time PCR system).
<Result>
The expression level of Bmal1 gene is shown in FIG. 1, and the expression level of Per1 gene is shown in FIG. As described above, it is known that the expression levels of Bmal1 and Per1 are in reverse phase in normal circadian rhythm. In this study, it was confirmed that the expression levels of Bmal1 and Per1 were in reverse phase when oleacein was added. Specifically, oleacein increased the expression level of Bmal1 while decreasing the expression level of Per1. In other words, it was found that oleacein has the effect of promoting the synchronization of the shifted phase and returning the circadian rhythm to normal by regulating the expression of the clock gene in a normal pattern.

Test Example 2 Circadian rhythm adjustment effect of oleacein high content olive extract The circadian rhythm adjustment effect of oleacein high content olive extract was evaluated by the following method using the expression of clock genes as an index.
<Test sample>
As the olive extract, the extract extracted in the steps shown in the following (1) and (2) was used. In addition, it is also possible to use Indena's Orea Select (trademark) for the extract.

(1) The residue obtained by squeezing olive oil from olive fruit was defatted with hexane and extracted with hot water at 80 ° C. for 30 minutes.
(2) The extract obtained in (1) was loaded onto an Amberlite XAD7-HP column (organo), water (6 L), 15% ethanol / water (8.4 L), 30% ethanol / water (4. 8L), stepwise elution of 45% ethanol / water (3.6L), 60% ethanol / water (4.8L), and freeze-dry the 30% ethanol fraction and 45% ethanol fraction for evaluation. It was.

  The 45% ethanol fraction contained at least 30% by weight of olive polyphenol and 3.2% by weight of oleacein. The oleacein content of the 30% ethanol fraction was 0.44% by weight. Oleacein was quantified by the following method. HPLC: Shimadzu LC2010C) Develosil XG-C30-5 column (Nomura Chemical, 4.6 x 150 mm), mobile phase was 0.1% formic acid aqueous solution as solution A and 90% acetonitrile solution containing 0.1% formic acid as solution B. Using. Elution was carried out using a linear concentration gradient over 20 minutes (B solution 10% → B solution 45%), and then B solution 45% was used for 3 minutes (flow rate: always 1.0 ml / min).

Under this condition, the peak corresponding to oleacein elutes at 14.1 minutes (detection wavelength: 280 nm).
<Evaluation method>
After 8-week-old male Wistar rats were acclimated for 1 week, they were divided into a control diet group and a test diet group (30% ethanol fraction administration group, 45% ethanol fraction administration group) (5 mice in each group). Next, the 30% ethanol fraction and 45% ethanol fraction were dissolved in physiological saline containing 10% Tween 80 and 5% EtOH, and this was gavaged once a day for 3 days at 250 mg / 5 ml / kg. Administered. In the control diet group, 10% Tween 80 was orally administered by gavage once a day for 3 days at 250 mg / 5 ml / kg. Four hours after the final administration, blood was collected from the abdominal aorta and the liver was collected.

Total RNA was extracted from the collected liver by a conventional method, and the expression levels of Bmal1 and Per1, which are clock genes, were extracted using the DNA microarray method (Affymetrix (registered trademark) GeneChip (registered trademark) Rat Genome 230 2.0 Array). Was analyzed.
<Result>
The expression level of Bmal1 gene is shown in FIG. 3, and the expression level of Per1 gene is shown in FIG. Compared with the control group, the expression level of Bmal1 was significantly increased and the expression level of Per1 was significantly decreased in the 45% ethanol fraction administration group. As described above, it is known that the expression levels of Bmal1 and Per1 are in reverse phase in normal circadian rhythm. This time, in the 45% ethanol fraction administration group, the reverse phase of Bmal1 and Per1 expression fluctuation was confirmed. Specifically, the 45% ethanol fraction increased the expression level of Bmal1 while decreasing the expression level of Per1. In other words, it was found that the olive extract promotes the synchronization of the shifted phase by adjusting the expression of the clock gene to a normal pattern, and restores the circadian rhythm to normal, and has a circadian rhythm adjustment effect. .

  Since oleacein accounts for about 10% of the polyphenols in the olive extract, it is considered to be an active ingredient of the circadian rhythm control effect of the olive extract.

  The circadian rhythm adjusting agent of the present invention can effectively adjust the circadian rhythm by including oleacein as an active ingredient. Therefore, the circadian rhythm adjusting agent of the present invention can be effectively used for the prevention, alleviation, treatment, etc. of physical disorders and diseases caused by disturbance of circadian rhythm such as sleep disorder and jet lag. it can.

Claims (10)

A circadian rhythm regulator containing oleacein as an active ingredient. The circadian rhythm adjusting agent according to claim 1, wherein the expression levels of the Per1 gene and the Bmal1 gene are adjusted to an opposite phase, thereby synchronizing the expression rhythm of the Per gene and the Bmal gene with a normal phase. The circadian rhythm regulator according to claim 1 or 2, wherein oleacein is contained as an olive extract. The circadian rhythm regulator according to claim 3, wherein the olive extract contains 30% by weight or more of olive-derived polyphenols. The circadian rhythm regulator according to claim 3 or 4, wherein the olive extract is obtained by extracting olive fruit with water, ethanol, or a mixture thereof. The circadian day according to any one of claims 3 to 5, wherein the olive extract is a 40% to 50% ethanol extract fraction of the defatted residue of olive fruit extracted with hot water at 70 ° C to 100 ° C. Rhythm adjusting agent. Food / beverage products for circadian rhythm adjustment which added the circadian rhythm regulator of Claim 1 or 2. A pharmaceutical composition for preventing, alleviating or treating sleep disorders or jet lag caused by disturbance of circadian rhythm, comprising oleacein. The manufacturing method of the food-drinks or pharmaceutical composition for circadian rhythm adjustment characterized by including the process of mix | blending oleacein. Use of oleacein in the manufacture of food or drink or pharmaceutical compositions for circadian rhythm adjustment .

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