KR20190061737A - Composition for Preventing, Improving or Treating Sleep Disorder Comprising Passiflora caerulea extract as an Active Ingredients - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating sleep disorders containing a Passiflora caerulea extract as an active component. According to the present invention, it is confirmed that the Passiflora caerulea extract increases the amount of c-fos expression in mammillary bodies and hippocampal tissues, and also promotes nerve regeneration in the hippocampus, thereby being able to prevent, alleviate or treat sleep disorders.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing, ameliorating or treating sleep disorders, which comprises a clock flower extract as an active ingredient.

The present invention relates to a composition for preventing, ameliorating or treating a sleep disorder, which comprises a clock flower extract as an active ingredient.

Sleep disturbance is one of the common problems that modern people are experiencing. Sleep disorder is a condition that does not take a healthy sleep, does not maintain awakening during the day despite having sufficient sleep, or is in trouble when sleeping or awake due to disordered sleep rhythm. It means.

As such, sleep disorder is a major cause of mental problems, but it can be caused by excessive stress, anxiety, tension, and fear. Sleep disorder is a fairly common disease that has been or has been experienced by more than 20% of the population and can lead to a variety of personal and social problems, including learning disabilities, inefficiencies, traffic accidents, safety accidents, emotional disturbances, Dissatisfaction with marriage, and industrial accidents. In addition, if the sleep disorder is not properly treated, the medical, neurological and psychiatric illnesses already suffering may be worsened or the recovery may be delayed, and serious diseases such as myocardial infarction and stroke may be caused. There is a growing interest in sleep aids.

Benzodiazepines and serotonin antagonists have been used as therapeutic agents for sleep disorders. Many drugs, including benzodiazepines, bind to GABAA receptors and perform pharmacological actions. When a drug or adjuvant is bound to the benzodiazepine site, the GABAA receptor affinity for GABA is enhanced, and the intracellular influx of chlorine ions To improve anxiety, to improve seizure, to sedation, and to induce and improve sleep. The GABAA receptor is a pentameric protein that forms a membrane ion channel and is closely related to the regulation of sedation, sleep, anxiety, muscle tension, convulsions, memory loss, etc. through which GABA (gamma-aminobutyric acid) acts. These drugs, when used for a long period of time, are accompanied by side effects that result in resistance and dependence, and may cause serious problems such as muscle relaxation and forgetfulness.

Therefore, the need for natural supplements is increasing as a means of replacing sleeping pills in the case of long-term sleepers and restrictors, and the demand for functional foods for sleep enhancement is increasing.

In this regard, Korean Patent No. 10-0947353 provides a health food composition for improvement of sleeping disorder and a pleasant surface. Especially, it is a ring composed of a sanjoin, a tranquilizer, a livestock meat, a lotus root and magnesium, Discloses a health food composition for improvement of sleeping disorder and a pleasant surface that exhibits excellent sleeping disorder improving effect by increasing the content.

Korean Patent Laid-Open Publication No. 10-2014-0030454 also discloses a fermentation material for relieving stress or improving sleeping disturbance containing a natural gas and fermenting a mixture of oysters and seaweeds, and a method for producing the same, Which contains a large amount of natural GABA, and a method for producing the fermented material.

The present inventors have sought to develop natural products capable of preventing or treating sleep disorders. As a result, the present inventors have found that clock flower (Passiflora caerulea) extracts the seed Force (c-fos) increase the expression level, increase the expression of the DCX protein in the hippocampus, by promoting nerve regeneration in brain tissue, sleep disorder, and improvement in the nipple body and the hippocampus tissue The present invention has been completed.

Accordingly, it is an object of the present invention to provide a composition for preventing, ameliorating or treating sleep disorders.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the present invention provides clock flower (Passiflora The present invention also provides a pharmaceutical composition for preventing, ameliorating, or treating sleep disorders, which comprises an extract of Corynebacterium sp .

In one embodiment of the present invention, the extract is an extract extracted with one kind of solvent selected from the group consisting of water, ethanol, dichloromethane, ethyl acetate, and butanol.

In another embodiment of the present invention, the clock flower extract is a leaf or fruit extract of a clock flower.

In another embodiment of the present invention, the extract is characterized by increasing the expression of C-fos and Doublecortin (DCX) proteins in hippocampal tissue.

In another embodiment of the present invention, the extract is characterized in that the expression of C-fos protein is increased in the hypothalamic papillary body tissue.

The present invention also relates to a method for producing The present invention provides a health functional food composition for improving sleeping disorder, which comprises, as an active ingredient,

The invention watch flower (Passiflora caerulea ), the expression of C-fos and DCX protein in hippocampal tissues was increased, the expression of C-fos protein was increased in the hypothalamic tissue of the hypothalamus, It is expected that the composition containing the clock flower extract of the present invention will exhibit an excellent treatment, prevention and improvement effect in sleep disorder related diseases.

FIG. 1 is a graph showing the effect of the clock flower extract of the present invention on intracellular ROS production and viability of C6 glioma cells.
FIG. 2 is a diagram showing the results of confirming the effect of oral administration of a clock flower extract on the mammillary body tissue of the hypothalamus of the mouse.
FIG. 3 is a graph showing the results of confirming the increase of c-fos protein in rat hippocampus tissue by administration of clock flower extract.
FIG. 4 is a graph showing the results of confirming the increase of DCX protein, which is an index of nerve regeneration, in rat hippocampus tissue by administration of clock flower extract.
FIG. 5 is a graph showing the results of confirming that the clock flower extract can enhance the learning effect of DBA / 2 mouse through water maze test.

The present inventors have made intensive efforts to develop a natural product with improved sleep disorder prevention or therapeutic effect, and as a result, confirmed that the extract of Passiflora caerulea has a regenerating effect on nerve cells in brain tissue, thereby completing the present invention.

That is, the present invention provides clock flower (Passiflora The present invention also provides a pharmaceutical composition for preventing, ameliorating, or treating sleep disorders, which comprises an extract of Corynebacterium sp .

The present invention also relates to a method for producing The present invention provides a health functional food composition for improving sleeping disorder, which comprises, as an active ingredient,

The clock flower extract used in the composition of the present invention is an extract obtained by adding a solvent to a clock flower flower, a leaf or a fruit, and the extract is a mixture of water, ethanol, dichloromethane, ethyl acetate, butanol, , And preferably may be a water (hot water) extract.

The extract is obtained by treating an extractive solvent in a clock flower, and various extraction solvents can be used. According to one embodiment of the present invention, a polar solvent or a non-polar solvent can be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, butanol, 2,2,4-trimethylpentane, decane, cyclobutanol, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o -xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF. Preferably, water or ethanol may be used as in the embodiment of the present invention. The clock flower extract used in the present invention may be prepared in powder form by an additional process such as vacuum distillation, freeze drying, or spray drying.

As used herein, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve efficacy or activity of the timepiece flower extracts described below. The present invention is an extract extracted from a natural flower material ( Passiflora caerulea ), and there is no adverse effect on the human body even when it is administered in an excessive amount. Therefore, the quantitative upper limit in which the clock flower extract is included in the composition of the present invention is selected by a person skilled in the art .

The composition of the present invention is effective for preventing, ameliorating, or treating sleep disorders related to dysfunction of hypothalamus and hippocampal tissues such as brain dysfunction. The composition of C-fos protein Lt; RTI ID = 0.0 > DCX < / RTI > protein in hippocampal tissues.

The pharmaceutical composition of the present invention comprises (a) a pharmaceutically effective amount of the above-mentioned clock flower extract of the present invention; And (b) a pharmaceutically acceptable carrier. As used herein, the term " pharmaceutically effective amount " means an amount sufficient to achieve efficacy or activity of the clock flower extract described above.

When the composition of the present invention is prepared from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components.

The pharmaceutical composition of the present invention can be administered orally or parenterally. According to one embodiment of the invention, it is applied in an oral administration mode.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on such factors as formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate, . Typical dosages of the pharmaceutical compositions of the invention are in the range of 0.001-100 mg / kg on an adult basis.

The pharmaceutical composition of the present invention may be formulated into a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The composition of the present invention can be provided as a food composition. When a composition for preventing, ameliorating or treating sleep disorder comprising the clock flower extract of the present invention as an active ingredient is prepared from a food composition, not only clock flower extract as an active ingredient but also ingredients normally added in food production And includes, for example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, .

The composition for preventing, ameliorating or treating sleep disorder comprising the clock flower extract of the present invention as an active ingredient may be prepared from a health functional food composition. When the composition of the present invention is prepared with a health functional food composition, it includes components that are conventionally added in the manufacture of foods, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, in the case of being made with a drink, a flavoring agent or a natural carbohydrate may be added as an additional ingredient in addition to the clock flower extract as an active ingredient. For example, natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.); Disaccharides (e.g., maltose, sucrose, etc.); oligosaccharide; Polysaccharides (e.g., dextrin, cyclodextrin and the like); And sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). Natural flavoring agents (e.g., tau marin, stevia extract, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) may be used as flavorings.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

[Example]

Example 1 Effective Fractionation and Effective Component Separation from Clock Flower

1.1. Preparation of extract

Passiflora caerulea was purchased and water or alcohol was added to make watch flower water or alcoholic extract. The water or alcohol extract was used in the following experiment.

Example 2. Confirmation of Effect of Clock Flower Extract on Cranial Cells

The efficacy was evaluated using C6 glioma cell, a type of neuronal cell, and the antioxidant effect of the clock flower extract was evaluated together with the fact that the sedative and sleeping inducing effect of the clock flower extract acted in the cranial nerve.

As a result, as shown in Fig. 1, it was confirmed that the clock flower extract did not impair viability of the C6 glioma cell at a proper concentration, not at a high concentration (Figs. 1A and 1C) (Fig. 1B).

Example 3. Confirmation of Increase of c-fos Expression Effect of Clock Flower Extract

The increase in C-fos protein in the brain tissue means that the stimulation is increased in the corresponding neuron where the expression is increased. In Example 3, the expression of C-fos protein in the brain tissue after oral administration of the clock flower extract .

It is known that the increase of c-fos protein expression in brain tissue is influenced by the clock flower material. It is known that the expression of C-fos protein by external stimulation in brain tissue appears within 15 minutes.

3.1. Increased expression of c-fos protein in papillary bodies

In the mammalian brain, the mammillary body is a tissue associated with sleep. In a previous study, it has been reported that shrinkage of the mammillary body occurs in patients with insomnia.

Then, 30 minutes after oral administration of high-concentration (500 mg / ml) clock flower extract (PF) to mice, mouse brain tissue was extracted and subjected to immunohistochemistry using C-fos antibody.

C-fos protein was significantly increased in the mammillary body of the hypothalamus. This implies that the clock flower extract acts on the hypothalamus, particularly the function of the mammillary body, which is associated with sleeping action.

As a result, as shown in Fig. 2, it was confirmed that the oral administration of the clock flower extract affects the mammillary body tissue of the hypothalamus of the mouse, thereby increasing the expression of c-fos protein.

3.2. Increased expression of c-fos protein in hippocampus

The present inventors have previously found that short-term administration of clock flower extract promotes nerve regeneration in the hippocampus of DBA / 2 mouse through immunohistochemistry of brain tissues. Thus, in Example 3.2, The effects on the hippocampus and hypothalamus were confirmed using western blot.

As described above, the increase of the C-fos protein is indirectly used as an indicator indirectly indicating that the electrical signal is continuously transmitted to the nerve cell. As shown in Fig. 3, when the clock flower extract is administered for a short time, Of the C-fos protein increased in the hippocampus of the hippocampus, indicating that the clock flower extract influenced the nerve cells of the hippocampal tissue.

Example 4. Confirmation of Increase of DCX Protein Expression of Clock Flower Extract

Doublecortin (DCX) protein in brain tissue is a type of neuronal migration protein that is used as an indicator of nerve regeneration in brain tissue.

Thus, in Example 4, the expression of DCX protein in hippocampal tissues after oral administration of clock flower extract was increased.

As a result, as shown in Fig. 4, it was confirmed that the expression of DCX was further increased as compared with the control group in which the clock flower extract was not administered. That is, the clock flower extract of the present invention has an effect of improving the sleeping disorder.

Example 5 Confirmation of Improvement of Sleep Disorder by Water Maze Experiment of Clock Flower Extract

It is considered that it is necessary to change the concentration of the clock flower extract in various ranges in order to set the correct optimum concentration to induce the sleep. In Example 5, the DBA / 2 mouse was administered with a high concentration (500 mg / kg) For 2 weeks.

DBA / 2 mouse is a mouse type having a sleeping disorder. In a previous study conducted by the present inventors, immunohistochemistry test was performed to confirm that the low concentration (10 mg / kg) clock flower extract exhibits nerve regeneration effect of DBA / 2 mouse hippocampus have. This suggests that the clock flower extract is a result of the improvement of sleep disturbance in DBA / 2 mice. Therefore, water maze test was performed immediately before the end of the animal experiment to evaluate the effect of promoting hippocampal regeneration in DBA / And it was confirmed that it appeared as a behavioral result.

The following experiment was carried out by applying the experimental method to the bitterness described in the existing literature. SK-PC-B70M from Pulsatilla koreana improves scopolamine-induced < RTI ID = 0.0 > impairments of memory consolidation and spatial working memory. Brain Res. 12: pp. 254-259, 2007)

The Morris water-maze test was conducted to investigate whether alpha-pinene helps the spatial perception of experimental animals and the short- and long-term memory recovery to be. Experimental equipment consists of a round steel tank (120 cm in diameter, 45 cm in height) and a platform (10 cm in diameter, 30 cm in height). The height of the water in the water tank (temperature 22 ± 2 ° C) was up to 2 cm above the platform so that when the mouse sat on the escape platform, the body could come out of the water. In the experiment with water, the test animals were kept at the same level during the experiment period so that the environment was not changed because the animals were looking for the escape pods using the markers around the water tanks. The time taken for the animals to visit when they visited the escape pod for more than 20 seconds was defined as the escape latency, and the mean value obtained by performing this three times a day was the mean escape latency. The incubation latency was recorded by observing the computer program (Ethovision 3.1, Noduls, The Netherlands) by installing a camera on the ceiling above the tank. The experiment was carried out three times daily for 5 days, and the possibility of visiting the esophagus by chance was minimized by sequentially changing the position of putting the experimental animals in the water tank every time. If the experimental animal can not find the esophagus within 120 seconds, the escape latency is set to 120 seconds. When the animal is sitting on the esophagus, it is left for 20 seconds to remind the surrounding clues.

As a result, as shown in Fig. 5, it was confirmed that the mice of the group to which the clock flower extract was administered showed a much faster decrease in the time taken to visit the buoys in the water maze than in the control group. As described above, the clock flower extract has an effect of promoting nerve regeneration. From the above results, it can be expected that the clock flower extract of the present invention can improve the sleep disorder.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (10)

A pharmaceutical composition for preventing, ameliorating, or treating a sleep disorder, comprising an extract of Passiflora caerulea as an active ingredient.
The method according to claim 1,
Wherein the extract is extracted with at least one solvent selected from the group consisting of water, ethanol, dichloromethane, ethyl acetate, and butanol.
The method according to claim 1,
The pharmaceutical composition for preventing, ameliorating, or treating sleep disorder, wherein the clock flower extract is a leaf or fruit extract of a clock flower.
The method according to claim 1,
The pharmaceutical composition for preventing, ameliorating, or treating sleep disorders, wherein the extract increases the expression of the cortex and double-core protein in hippocampal tissue.
The method according to claim 1,
A pharmaceutical composition for preventing, ameliorating, or treating a sleep disorder, wherein the extract increases the expression of the c-spos protein in the hypothalamic papillary body tissue.
A health functional food composition for improving sleeping disorder, comprising Passiflora caerulea extract as an active ingredient.
The method according to claim 6,
Wherein the extract is extracted with at least one solvent selected from the group consisting of water, ethanol, dichloromethane, ethyl acetate, and butanol.
The method according to claim 6,
Wherein the clock flower extract is a leaf or fruit extract of a clock flower.
The method according to claim 6,
Wherein said extract enhances the expression of the cortex and double core protein in hippocampal tissue.
The method according to claim 6,
Wherein said extract increases the expression of c-spos protein in the hypothalamic tissue of the hypothalamus.

Images