KR102240706B1 - A composition for sleep induction comprising Passiflora incarnata extract - Google Patents

Abstract

The present invention relates to a composition for inducing sleep. By containing Passionflower (Passiflora incarnata) extract as an active ingredient, GABA nervous system activity and blood melatonin are improved while other body composition is not changed, so it is excellent in inducing sleep without side effects of metabolism. Do.

Description

A composition for sleep induction comprising Passiflora incarnata extract}

The present invention relates to a composition excellent in inducing sleep without side effects of metabolism by containing the Passion Flower (Passiflora incarnata) extract as an active ingredient.

Sleep, one of the important physiological functions of the human body, is essential for maintaining basic human functions, and is a process that recovers damage to the brain or body. Sleep has a great influence on the quality of life and function, and it is reported that about 30% of all patients suffer from sleep disorders. In particular, sleep is closely related to the immune system.

The central nervous system (CNS) regulates immune function by sending signals to target cells in the immune system through the autonomic nervous system and neuroendocrine pathways. Neurotransmitters and hormones produced and released by these pathways interact with immune cells to produce cytokines, and the produced cytokines regulate sleep. In addition, sleep disorders cause decreased motivation and mental fatigue, cause health problems including psychosocial phenomena such as concentration disorders and neurophysiological phenomena, and adversely affect overall disease recovery.

To treat these sleep disorders, patients are taking various sleeping pills. Hypnotics include the benzodiazepine series and zolpidem, which contain ingredients such as triazolam, lorazepam, and diazepam. Doxepin), trazodone, and other components are divided into antidepressants.

However, taking these sleeping pills can be addictive, and there are side effects such as appetite disorder, weight gain, and abnormalities of the autonomic nervous system when taken for a long time, requiring regulation, and neuropsychiatric abnormal behaviors and symptoms such as depression, anxiety and suicide. It has been reported in numerous studies. In other words, very dangerous physical and mental changes have been reported due to sleeping pills.

Therefore, there is a need for a sleep inducing agent made of natural materials that can improve sleep without side effects of metabolism.

Korean Patent Registration No. 1918249 Republic of Korea Patent Publication No. 2019-0017860

An object of the present invention is to provide a composition capable of inducing sleep without side effects of metabolism by containing an extract of Passionflower (Passiflora incarnata) as an active ingredient.

In addition, another object of the present invention is to provide a composition for inducing secretion of melatonin containing an extract of Passiflora incarnata as an active ingredient.

The composition for inducing sleep of the present invention for achieving the above object may contain an extract of Passionflower (Passiflora incarnata) as an active ingredient.

The passionflower extract may be a mixture of passionflower leaves and passionflower fruit in a weight ratio of 1:0.1-0.8.

The passion flower extract may be extracted by mixing a passion flower, water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof in a volume ratio of 1:1 to 100.

The mixed solvent may be an aqueous solution of 20 to 80% by volume of methanol, ethanol, butanol or propanol.

The passion flower extract may be used at 500 to 1500 mg/kg.

In addition, the composition for inducing melatonin secretion of the present invention for achieving the above-described other object may contain Passionflower (Passiflora incarnata) extract as an active ingredient.

The composition for inducing sleep containing the passionflower extract of the present invention as an active ingredient has an effect on the secretion of seratonin and melatonin, and has an effect on inducing sleep, so that it can be used as a functional food or pharmaceutical use.

1A is a Western blot showing the expression of caletinine in the hippocampus and hypothalamus separated from the PI 700 group and the control group to which the PI extract and distilled water were repeatedly administered, and FIG. 1B is a graph showing the caletinine expression in the hippocampus of FIG. 1C is a graph showing the expression of caletinine in the hypothalamus of FIG. 1A in numerical terms, and FIG. 1D is a graph showing the change in body weight during administration of the extract to the PI 700 group and the control group (5 days), and FIG. 1E Is a graph measuring the level of melamine, a sleep-related hormone, after administration of the extract repeatedly for 5 days, and FIG. 1F is a graph measuring the level of serotonin, a sleep-related hormone, after administration of the extract repeatedly for 5 days.
Figure 2A is a graph showing the amount of the PI 700 group and the control group to which the PI extract and distilled water were repeatedly administered and the control group consumed the diet for 24 hours, and FIG. 2B is that the PI 700 group and the control group consumed the diet during the dark cycle and 24 hours. It is a graph showing the amount, Figure 2C is a graph showing the amount of water intake of the PI 700 group and the control group for 24 hours, Figure 2D is the amount of water intake of the PI 700 group and the control group during the dark cycle and 24 hours It is a graph showing.
Figure 3A is a graph measuring RER for 24 hours of the PI 700 group and the control group repeatedly administered PI extract and distilled water, and Figure 3B is a graph measuring the dark cycle of the PI 700 group and the control group and RER for 24 hours, Figure 3C is a graph measuring EE for 24 hours in the PI 700 group and the control group, Figure 3D is a graph measuring the dark cycle of the PI 700 group and the control group and EE for 24 hours, Figure 3E is the PI 700 group And a graph measuring the activity for 24 hours of the control group, and FIG. 3F is a graph measuring the dark cycle and the activity for 24 hours of the PI 700 group and the control group.
Figure 4A is a graph showing the mass by measuring the fat, body fluids and lean of the PI 700 group and the control group to which the PI extract and distilled water were repeatedly administered, and FIG. 4B is a PI 700 group to which the PI extract and distilled water were repeatedly administered, and It is a graph showing fat, body fluid, and lean in the control group as a composition ratio.
5 is a graph measured by HPLC of the PI extract extracted according to Example 1 of the present invention.

The present invention is to provide an excellent composition for inducing sleep without metabolic side effects by containing the Passion Flower (Passiflora incarnata) extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The composition for inducing sleep of the present invention contains Passion Flower (Passiflora incarnata) extract as an active ingredient.

The Passion Flower (Passiflora incarnata) is an evergreen perennial vine plant and may use Passion Flower flowers, leaves, stems, roots, and fruits. Native Americans used whole plants for swollen or irritated eyes, and the roots are commonly used as a tonic. In addition, leaves prevent rapid pulse, reduce high blood pressure, are non-toxic, and are used as anxiety sedatives, as well as relieve asthma muscle cramps, epilepsy, nervous irritability, and colonic syndrome, and are used as poultices to soften burns and skin inflammation.

Passion flower extract of the present invention is an extract in which passionflower leaves and passionflower fruits are mixed in a weight ratio of 1:0.1-0.8, preferably 1:0.2-0.5. When the content of the passionflower fruit is less than the lower limit based on the passionflower leaf, the sleep-inducing effect cannot be expected, and when the content of the passionflower fruit exceeds the upper limit, the effect of the present invention may not be expressed at all and toxicity may be induced.

When the passionflower leaves and passionflower fruits are not used together, but used alone, the efficacy is 1.5 to 20 times lower than that of using the two substances together; When passionflower flowers, stems, and roots are used, the efficacy is 5 to 50 times lower than when the two substances are used together.

The mixture of passionflower, for example passionflower leaves and passionflower fruit, is mixed with an extraction solvent in a weight ratio of 1: 1 to 100, preferably 1: 5 to 20, and extracted at 70 to 100°C to prepare an extract. When the weight ratio of the mixture of the passionflower leaf and the passionflower fruit and the extraction solvent is out of the above range, the active ingredient of the passionflower leaf and passionflower fruit mixture may be extracted in a small amount in the extract.

The extraction solvent for extracting the extract is water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The extraction solvent is not particularly limited, but an extract extracted with 20 to 80 vol% of methanol, ethanol, butanol or propanol aqueous solution, preferably 20 to 80 vol% of ethanol exhibits excellent effects.

The term'extract' used in referring to the passion flower in the present specification includes not only the crude extract obtained by treating the extraction solvent, but also the processed product of the passion flower extract. For example, the passion flower extract may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

In addition, the passion flower extract of the present invention has a meaning in a broad sense to include a processed product of a passion flower, such as a passion flower powder, formulated so that the passion flower can be administered to an animal. Although the experiment was conducted with the passion flower extract in the present invention, it will be foreseen by those skilled in the art that the desired effect can be achieved even in the same form as the processed product of the passion flower.

Meanwhile, in the present specification, the term "contained as an active ingredient" means including an amount sufficient to achieve the efficacy or activity of the passionflower extract. For example, the passion flower extract is used in a concentration of 500 to 1500 mg/kg, preferably 600 to 1000 mg/kg. Passion flower extract is a natural product and does not have side effects on the human body even if it is administered in an excessive amount. Therefore, the upper limit of the quantity of the passion flower extract contained in the composition of the present invention can be selected and carried out by a person skilled in the art within an appropriate range.

The composition of the present invention may be used as a pharmaceutical composition for inducing sleep, and may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant may include excipients, disintegrants, Sweetening agents, binders, coating agents, expanding agents, lubricants, lubricants or flavoring agents may be used.

For administration, the pharmaceutical composition may contain one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, trackcacanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

Acceptable pharmaceutical carriers for compositions formulated as liquid solutions are sterilized and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injection formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Further, it can be preferably formulated according to each disease or ingredient using a method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration. .

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, Usually the skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention may be prepared in unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient, or may be prepared by incorporating it into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

In addition, the present invention provides a food composition for inducing sleep containing the passionflower extract as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice creams, vitamin complexes, health supplements. Etc.

The food composition of the present invention may include not only the passionflower extract as an active ingredient, but also ingredients commonly added during food production, and, for example, include proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents. . Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, in addition to the passion flower extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included. .

The present invention provides a health functional food comprising a food composition for inducing sleep comprising the passion flower extract as an active ingredient. Health functional foods are foods prepared by adding passionflower extract to food materials such as beverages, teas, spices, gums, confectionery, or encapsulated, powdered, or suspension. However, unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of passionflower extract added in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but can be added within the range that does not damage the original taste of the food. It is in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, the present invention provides a use of the passionflower extract for the manufacture of a sleep-inducing medicine or food. As described above, the passion flower extract can be used for inducing sleep.

In addition, the present invention provides a method for inducing sleep comprising administering an effective amount of passionflower extract to a mammal.

The term "mammal" as used herein refers to a mammal that is an object of treatment, observation or experiment, and preferably refers to a human.

The term "effective amount" as used herein refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, which Includes an amount that induces relief of symptoms of the disease or disorder. The effective amount and frequency of administration for the active ingredient of the present invention may be changed according to the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. It can be adjusted according to various factors including condition, sex and diet, time of administration, route of administration and rate of secretion of the composition, duration of treatment, and drugs used concurrently. In the prophylaxis, treatment or improvement method of the present invention, in the case of an adult, it is preferable to administer the passionflower extract at a dose of 500 to 1500 mg/kg when administered once to several times a day.

In the treatment method of the present invention, the composition comprising the passionflower extract as an active ingredient is a conventional method through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered as.

Hereinafter, a preferred embodiment is presented to aid in the understanding of the present invention, but it is obvious to those skilled in the art that various changes and modifications are possible within the scope of the present invention and the scope of the technical idea, but the following examples are only illustrative of the present invention, It is natural that such modifications and modifications fall within the scope of the appended claims.

Example 1. Passion flower leaves: Passion fruit = 1: 0.25 weight ratio extract (PI extract)

A mixture of passionflower leaves and passionflower fruits in a weight ratio of 1: 0.25 was mixed with 60% ethanol, extracted for 4 hours at 80°C, filtered, concentrated, and spray dried to obtain passionflower extract (PI extract). (Fig. 5).

<Test Example>

Mouse preparation

Many studies are being conducted to verify the effect as a composition for inducing sleep, but most of the studies are using animals because there are limitations in studies involving humans. The test was performed using C57BL/6 mice purchased from Charles River (Incheon, Korea) in order to investigate how the administration of the Passion Flower extract of Example 1 affects feeding, activity, or metabolic changes.

As the experimental animals, 10 7-week-old C57BL/6 male mice (average weight of 22 g) were purchased.

After acclimation for 1 week before the start of the experiment, they were reared at room temperature (22±1 ℃) and 60% humidity under a 12 hour light cycle (light cycle from 7:00-19:00). The animals were provided to freely use normal food (2018S; Harlan, USA) and water, and distilled water or Passionate extract was given at around 17:00, 2 hours before the daily turn-off for 5 days from the start of administration (day 1). It was administered orally.

All test substances used in this experiment were administered orally at the same time every day, and all procedures related to the mouse were performed according to the ethical standards of the Institutional Animal Care and Use Committee (IACUC approval no. SCH16-0037) of Soonchunhyang University.

-2 groups of mice-

Control (Veh): distilled water (DW) 700 mg/kg _weight /day 5 mice in the diet group

PI 700 group (PI 700): Passion flower extract of Example 1 700 mg/kg _Weight /day 5 dietary groups

The mice were anesthetized with 1 g/kg of urethane (Sigma-Aldrich, St. Louis, MO, USA) on the 5th day of administration, and perfused with 0.1 M phosphate buffer (pH 7.4) to remove as much blood as possible. Before perfusion, blood was collected from the abdominal vein to measure serum serotonin and melatonin. After completely emptying the blood vessels, the brain was removed and the hypothalamus was separated, and the brain tissue was stored at -80°C for later processing. For the experiment, it was monitored with Phenomaster.

Western blot

The separated hippocampus and hypothalamus were homogenized in a lysis buffer (iNtRon Biotechnology, Seoul, Korea), and the protein concentration was measured with a BCA kit (iNtRon Biotechnology). Total protein (20 μg per sample) was placed in each lane of 12% SDS-PAGE, subjected to electrophoresis, and then transferred to a PVDF membrane (Bio-Rad Laboratories, CA, USA).

Thereafter, the membrane was blocked with TBST [100 mM Tris-HCl (pH 7.6), 0.8% NaCl and 0.1% Tween-20] containing 10% skim milk (BD Biosciences, CA, USA) and the membrane was blocked with the following 1 Incubated with primary antibody. Incubation was performed overnight at 4 °C with rabbit anti-calretinin (1:3000, Swant, Switzerland) and rabbit anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase, 1:5000, Cell Signaling Technology, MA, USA).

After culturing is complete, the membrane is incubated with horseradish peroxidase (HRP)-conjugated anti-rabbit secondary antibody (Vector, CA, USA), and the immune response signal is enhanced through chemiluminescence (Abclon, Seoul, Korea). Was detected and recorded with the MicroChemi 4.2 system.

Monitoring behavioral and metabolic changes by indirect calorimetry

Metabolic performance parameters (energy intake and energy expenditure) were studied using an indirect calorimeter system (TSE System GmBH, Bad Homburg, Germany) incorporating Phenomaster. Before the experiment, mice were acclimated for 2 days in a metabolic chamber containing food and water, and then oxygen consumption (VO ₂ ), carbon dioxide production (VCO ₂ ), and food consumption were measured for 7 days. The respiratory rate (RER; VCO ₂ /VO ₂ ) was calculated using standard in-house software. Energy consumption (EE) was calculated by the following [Equation 2].

[Equation 2]

EE = 3.815 X 10 ^-3 X VO ₂ + 1.232 X 10 ^-3 X VCO ₂

Body composition (tissue, fat and body fluids of mice on a bench top platform) was measured using a Mini-spec LF50 (Bruker Biospins, The Woodlands, TX).

HPLC analysis

Passion flower extract powder (1 g) of Example 1 was dissolved in 50 ml of a 50% ethanol aqueous solution, subjected to ultrasonic treatment for 10 minutes, and then filtered through a 0.45 μm syringe filter. HPLC was performed using an Agilent 1200 system equipped with a model G1312A binary LC pump, auto sampler and diode array detector. C-18 (Waters, SunFire TM C18) column (250 mm X 4.6 mm id and 5 μm particle size) was selected, and the standard was purchased from Sigma Chemicals (St. Louis, MO, USA).

Chromatographic separation was performed with a mobile phase composed of 0.1% phosphoric acid prepared in nanopure water (87%) and 100% acetonitrile (13%) for 60 minutes, and the injection volume was 20 μL. The mobile phase flow rate is 1 ml min-1, the oven temperature is 35° C., and the detection wavelength is 360 nm.

Data analysis

In order to ensure objectivity, two observers per experiment under the same conditions performed all conditions as blind conditions. The intensity of the band generated during western blotting was evaluated based on the measured optical density by converting the average gray level using the following [Equation 1] (using NIH Image 1.59 software).

[Equation 1]

Optical density = log (256/average gray level)

Data shown represent the experimental mean ± standard error (SE) for each experimental group. Differences between means were analyzed by one-way ANOVA, where appropriate, t-test and/or unpaired t-test.

Post-hoc analysis was performed by Fisher's most important difference, but Newman-Keul test, and statistical analysis was performed using JMP9 (SAS Institute, USA) and Prism7 (GraphPad Software, San Diego, CA, USA). P values (*: <0.05, **: <0.005, ***: <0.0005) were considered statistically significant.

Test Example 1. GABA activation in the brain by repeated PI administration, measurement of body weight and hormone changes

1A is a Western blot showing the expression of caletinine in the hippocampus and hypothalamus isolated from the PI 700 group and the control group, FIG. 1B is a graph showing the caletinine expression in the hippocampus of FIG. 1A in numerical terms, and FIG. 1C is the FIG. It is a graph showing the expression of caletinine in the hypothalamus as a numerical value, and FIG. 1D is a graph showing the change in body weight during administration of the extract to the PI 700 group and the control group (5 days), and FIG. 1E is a repeated extract of the extract for 5 days. It is a graph measuring the level of melamine, a sleep-related hormone, after administration, and FIG. 1F is a graph measuring the level of serotonin, a sleep-related hormone, after administration of the extract repeatedly for 5 days.

As shown in FIGS. 1A to 1C, it was confirmed that the expression of caletinin in the hippocampus and hypothalamus of the PI 700 group was significantly higher than that in the hippocampus and hypothalamus of the control group.

That is, the PI extract prepared according to Example 1 of the present invention increases calretinin in the hippocampus and hypothalamus known as calcium binding proteins secreted by GABA neurons.

In addition, as shown in FIG. 1D, it was confirmed that the change in body weight of the PI 700 group to which the extract of Example 1 was repeatedly administered was similar to that of the control group during the 5 days of repeated administration.

In addition, as shown in Figs. 1E and 1F, it was confirmed that the serum melatonin of the PI 700 group was significantly higher than that of the control group, and the serum serotonin level was slightly higher in the PI 700 group than that of the control group. Parvalbumin is known as a marker of the GABA nervous system, and parvalbumin protein expression was higher in the hippocampus of mice administered with PI extract than in the control group (data not shown).

Serotonin is a kind of catecholamine, plays a very important role as a wake-cycler, and is a major regulator of melatonin secretion. Serotonin stimulates the secretion of melatonin, but it also plays a role in sleeping.

That is, serotonin tended to increase in the PI 700 group, but was not statistically significant compared to the control group, and melatonin was significantly increased in the PI 700 group compared to the control group.

On the other hand, from the time the blood was sampled until 2 hours before the light was turned off, the serotonin level decreased, the melatonin stimulation increased instead, and the increased melatonin improved the immune response by reducing the level of superoxide anion radicals produced by heterogeneous organisms Let it.

Test Example 2. Measurement of food intake and water consumption

Food intake and water consumption were recorded for 5 days by Phenomaster® metabolic cages. All data obtained from the Phenomaster® metabolic cage was modified to provide real-time average consumption over a 24-hour light and dark cycle.

Figure 2A is a graph showing the amount of the PI 700 group and the control group consumed diet for 24 hours, Figure 2B is a graph showing the amount of the PI 700 group and the control group consumed diet during the dark cycle and 24 hours, Figure 2C is The PI 700 group and the control group are graphs showing the amount of water ingested for 24 hours, and FIG. 2D is a graph showing the amount of the PI 700 group and the control group consumed water during the dark cycle and 24 hours. ZT stands for Zeitgeber time.

2A and 2C, PI extract and distilled water were administered 2 hours before the light was turned off, and the diet and water intake of the PI 700 group significantly decreased 2-3 hours before the light was turned on. It was confirmed that there was no difference.

In addition, as shown in FIGS. 2B and 2D, diet intake and water consumption during each dark cycle increased in both groups, but no difference in volume was found between the two groups. Moreover, it was confirmed that the total consumption for each dark cycle and the total 24 hours did not differ from each other.

Test Example 3. Measurement of changes in RER, EE and activity by repeated administration of PI extract

Figure 3A is a graph measuring RER for 24 hours of the PI 700 group and the control group, Figure 3B is a graph measuring the dark cycle of the PI 700 group and the control group and RER for 24 hours, and Figure 3C is a graph of the PI 700 group and the control group. It is a graph measuring EE for 24 hours, FIG. 3D is a graph measuring the dark cycle of the PI 700 group and the control group and EE during 24 hours, and FIG. 3E is a graph measuring the activity of the PI 700 group and the control group for 24 hours. , Figure 3F is a graph measuring the activity during the dark cycle and 24 hours of the PI 700 group and the control group. PI extract and distilled water were administered 2 hours before the light was turned off.

As shown in Figs. 3A and 3B, the RER (VCO ₂ /VO ₂ ) value of the PI 700 group was high during the light cycle, and it was confirmed that it was higher for about 11 hours after oral administration of the PI extract. However, from 04:00 to 06:00, the pattern of RER was reversed and the RER of the control group started to rise. In the RER real-time graph, only significant 3 time-points appeared, but it was confirmed that the AUC of accumulated RER for the dark cycle and the total 24 hours was not significant.

In general, energy consumption (EE) is known to be about 15% less in the sleep state than in the awake state, and as shown in Figs. 3C and 3D of the present invention, the EE results are EE real-time (Fig. 3C) and the AUC of the EE. (Figure 3D) After administration of the PI extract in all, the PI 700 group decreased about 16% during the dark cycle. The result of accumulated EE showed a decrease of 8.8% in the PI 700 group over 24 hours.

In addition, as shown in Figs. 3E and 3F, after administration of the PI extract, the PI 700 group showed lower activity during the dark cycle in a real-time activity graph than the control group, and gradually increased from 2 hours before the light was turned on (red dot circle). After lighting, it was confirmed that it showed similar activity to the control group.

It was confirmed that the PI 700 group maintained low activity during the dark cycle, and the cumulative activity of the PI 700 group was considerably low.

Test Example 4. Measurement of changes in body composition by repeated administration of PI extract

Figure 4A is a graph showing the mass by measuring fat, body fluid and lean of the PI 700 group and the control group repeatedly administered with PI extract and distilled water, and FIG. 4B is a PI 700 group repeatedly administered with PI extract and distilled water, and It is a graph showing fat, body fluid, and lean in the control group as a composition ratio.

Immediately prior to sacrifice of the animal, the body composition of the mice was quickly measured. It is known that the obtained fat, body fluids and lean body generally account for about 95% of the total body weight.

As shown in Figs. 4A and 4B, it was confirmed that there was no difference in the contents of fat, body fluid, and lean between the PI 700 group and the control group. This data implies that repeated oral administration of PI extract did not change body composition.

These results of the present invention mean that the PI extract is effective in increasing GABA nervous system activity and blood melatonin levels, and it was confirmed that a significant decrease in EE was shown when mice were generally active in the dark cycle. That is, no increase in appetite or increase in weight was observed, and all body composition was not changed. Only sleep has changed.

In the present invention, the PI extract was repeatedly administered to mice for 5 days to find no abnormal metabolic phenotypes or side effects of behavior such as hyperactivity or unexpected metabolic changes, so it can be used as a sleep inducing agent without metabolic side effects.

Hereinafter, examples of the formulation of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit it, but is intended to be described in detail.

Formulation Example 1. Preparation of powder

500 mg of extract powder obtained in Example 1

100 mg lactose

10 mg of talc

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2. Preparation of tablets

300 mg of extract powder obtained in Example 1

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet manufacturing method.

Formulation Example 3. Preparation of Capsule

200 mg of extract powder obtained in Example 1

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

Formulation Example 4. Preparation of injection

600 mg of extract powder obtained in Example 1

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO _4, 12H ₂ O 26 mg

It is prepared with the above ingredients per ampoule according to a conventional injection preparation method.

Formulation Example 5. Preparation of liquid formulation

4 g of extract powder obtained in Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid formulations, add and dissolve each component in purified water, add an appropriate amount of lemon flavor, mix the above ingredients, add purified water and add purified water to adjust the total to 100 g, then fill in a brown bottle for sterilization. To prepare a liquid.

Formulation Example 6. Preparation of granules

1,000 mg of extract powder obtained in Example 1

The right amount of vitamin mixture

Vitamin A acetate 70 ㎍

1.0 mg of vitamin E

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

Vitamin B12 0.2 ㎍

10 mg of vitamin C

Biotin 10 ㎍

1.7 mg of nicotinic acid amide

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

90 mg of potassium citrate

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules, but it is possible to arbitrarily modify the mixing ratio. After mixing the above ingredients according to a conventional granule preparation method, the granules And, according to a conventional method, it can be used for preparing a health functional food composition.

Formulation Example 7. Preparation of functional beverage

1,000 mg of extract powder obtained in Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

900 mL total by adding purified water

After mixing the above ingredients according to the normal health drink manufacturing method, stirring and heating the mixture at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and then stored in a refrigerator. It is used to prepare the functional beverage composition of the present invention.

The composition ratio is a mixture of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

Claims (6)

Contains Passion Flower (Passiflora incarnata) extract as an active ingredient,
The passion flower extract is a composition for sleep induction, characterized in that the passion flower leaves and passion flower fruits are mixed in a weight ratio of 1: 0.1-0.8. delete The composition for inducing sleep according to claim 1, wherein the passion flower extract is extracted by mixing a passion flower, water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof in a volume ratio of 1:1 to 100. The composition for inducing sleep according to claim 3, wherein the mixed solvent is an aqueous solution of 20 to 80% by volume of methanol, ethanol, butanol or propanol. The composition for inducing sleep according to claim 1, wherein the passion flower extract is used in an amount of 500 to 1500 mg/kg. delete

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