Cognitive function and memory improvement composition comprising natural extract mixture, and use thereof

Abstract

The present invention relates to a cognitive function and memory improvement composition comprising a natural extract mixture, and a use thereof. The cognitive function or memory improvement composition comprising a Perilla frutescens var. acuta Kudo extract and a peppermint (Mentha piperita) extract, according to one embodiment of the present invention, is based on natural materials and has few side effects, is easily accessible, and has economic efficiency while exhibiting an excellent cognitive function improvement effect, and thus may be usefully employed for the prevention, mitigation, or treatment of cognitive impairment- or memory impairment-related diseases.

Description

Composition for improving cognitive function and memory comprising a natural extract mixture and use thereof

[0001] The present invention relates to a composition for improving cognitive function and memory comprising a natural extract mixture and its use.

[0002] This research was conducted with the support of the Gyeonggi-do Regional Cooperation Research Center (Project No. GRRCAjou2023-B01).

[0003] Cognitive impairment (CAD) refers to a condition in which cognitive function declines compared to that of a normal elderly person, but the individual is still able to function independently, making it an intermediate state that cannot be diagnosed as dementia. This condition carries a high risk of developing dementia. Cognitive decline is distinct from the natural processes of normal aging and can be severe enough to interfere with daily life.

[0004] Mild cognitive impairment (MCI) is an early stage at risk for developing dementia, characterized by cognitive decline that does not significantly impact daily life. MCI typically begins five years before the onset of dementia, and approximately 80% of individuals with MCI develop dementia after six years.

[0005] The causes of cognitive impairment are diverse, but cell death, which leads to nerve cell damage and loss, is known to be the ultimate cause. Reactive oxygen species, a type of highly oxidizing oxygen that damages cells, are identified as a major cause of nerve cell death, the leading cause of degenerative neurological disorders. Oxidative stress caused by reactive oxygen species causes irreversible damage to brain cells and alters the structure and function of mitochondria, leading to cognitive impairments such as Parkinson's disease and Alzheimer's disease.

[0006] As we enter a super-aging society, the incidence of neuropsychiatric disorders and Alzheimer's disease is increasing, and recently, 43.9% of the 2030 generation is suffering from 'young Alzheimer's', which refers to severe forgetfulness in young people.

[0007] To date, many patents related to compositions having effects of treating cognitive impairment, improving cognitive ability, or enhancing memory have been published and registered in Korea. Publication No. 2014-0144785 relates to a composition for enhancing memory and learning ability containing citron extract as an active ingredient, and registration No. 10-1823892 relates to an extract of balsam that exhibits effects of improving memory, improving cognitive ability, and preventing, delaying, or treating dementia.

[0008] Representative medications currently on the market for the treatment of cognitive impairment include cholinesterase inhibitors (donepezil, rivastigmine, galantamine) and NMDA receptor antagonists (memantine). Donepezil inhibits the breakdown of acetylcholine in the brain, increasing neurotransmitter concentrations, while memantine inhibits excessive stimulation of nerve cells, preventing cell damage. However, donepezil has been reported to cause side effects such as nausea, diarrhea, muscle cramps, and sleep disturbances, while memantine can also cause headaches, drowsiness, dizziness, and vomiting.

[0009] Therefore, the development of therapeutic agents using natural materials compatible with the human body is in progress, and there is also a need for the development of new materials with fewer side effects.

[0010] In order to solve the above problems, the inventors of the present invention completed the present invention by confirming the cognitive function and memory improvement efficacy of a natural extract mixture containing the above natural products based on the fact that natural products such as Perilla frutescensvar. acutakudo and peppermint (Mentha piperita) are easily accessible, economical, and effective in improving cognitive function.

[0011] Accordingly, the purpose of the present invention is to provide a composition for improving cognitive function or memory comprising a perilla frutescensvar. acutakudo extract and a peppermint (Mentha piperita) extract.

[0012] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating diseases related to cognitive dysfunction or memory impairment, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

[0013] Another object of the present invention is to provide a method for preventing or treating a disease related to cognitive dysfunction or memory impairment, comprising the step of administering the pharmaceutical composition to a subject.

[0014] Another object of the present invention is to provide a food composition for improving cognitive function or memory, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

[0015] Another object of the present invention is to provide a feed composition for improving cognitive function or memory, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

[0016] The terminology used herein is for the purpose of description only and should not be construed as limiting. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this specification, the terms "comprises" or "has" and the like are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0017] Additionally, unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0018] In the present invention, duplicate content or terms have been omitted to avoid excessive complexity of this specification.

[0019]

[0020] In order to achieve the above object, according to one aspect of the present invention, a composition for improving cognitive function or memory is provided, which includes a Perilla frutescensvar. acutakudo extract and a peppermint (Mentha piperita) extract.

[0021] The term "cognitive function" used in the present invention refers to various intellectual abilities such as attention, perception, memory, language ability, and executive ability, and may be used interchangeably with "cognitive ability" or "cognitive ability."

[0022] The term "memory" or "memory" used in the present invention refers to the ability to acquire new information, learned experiences, or knowledge obtained from the surrounding environment, encode and store it in a specific part of the brain, and recall it as a result of learning that persists over time.

[0023] In one embodiment of the present invention, the tea extract and peppermint extract may be extracted using water, an organic solvent, or a mixture thereof as an extraction solvent, preferably water or a C1 to C4 lower alcohol, more preferably water, methanol, ethanol, or propanol as an extraction solvent, and most preferably water as an extraction solvent at 70°C to 100°C.

[0024] In one embodiment of the present invention, the composition may be characterized in that the tea extract and the peppermint extract are mixed in a weight ratio of 1:0.5 to 1:4, and preferably, the tea extract and the peppermint extract may be mixed in a ratio of 1:1.

[0025]

[0026] According to another aspect of the present invention, a pharmaceutical composition for preventing or treating a disease related to cognitive dysfunction or memory impairment is provided, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

[0027] The term "cognitive dysfunction" used in the present invention is a concept that can include a state caused by a decline in cognitive function as described above, a disease resulting therefrom, and all diseases that have the condition as a symptom, and preferably can mean a disorder caused by damage to brain nerve cells.

[0028] The term "memory disorder" as used herein refers to a condition in which the ability to form, maintain, or recall memories, either temporarily or persistently, is impaired, including hypermnesia or amnesia. Memory disorders can develop when any one of the three processes of memory—recording, retention, and retrieval—is impaired.

[0029] In the present invention, the disease related to cognitive dysfunction or memory impairment may specifically be a degenerative brain nervous system disease, and more specifically, the disease related to cognitive dysfunction or memory impairment may mean a degenerative brain disease selected from the group consisting of mild cognitive impairment (MCI), Parkinson's disease, Alzheimer's disease, amnesia, dementia, Pick's disease, and Huntington's disease.

[0030] The term "prevention" used in the present invention means any action that suppresses or delays the onset of a disease related to cognitive dysfunction or memory impairment by the extract of the perilla frutescensvar. acutakudo or the extract of the peppermint (Mentha piperita) of the present invention.

[0031] The term "improvement" or "treatment" used in the present invention means any action that improves or benefits a parameter related to a disease related to cognitive dysfunction or memory impairment, such as the degree of symptoms, by the extract of Perilla frutescensvar. acutakudo or the extract of peppermint (Mentha piperita) of the present invention.

[0032] The pharmaceutical composition of the present invention may be administered orally or parenterally in various dosage forms. When formulating the composition, it may be prepared using one or more buffers (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g., aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrating agents, or surfactants, diluents, or excipients.

[0033] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing one or more compounds with at least one excipient, such as starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethyl-cellulose or gelatin. For example, tablets or sugar-coated tablets can be obtained by mixing an active ingredient with a solid excipient, grinding the mixture, adding a suitable auxiliary agent, and then processing the mixture into a granule mixture.

[0034] In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, or preservatives may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as disintegrants in some cases, and anticoagulants, flavoring agents, emulsifiers, solubilizers, dispersants, flavoring agents, antioxidants, packaging agents, pigments, and preservatives may be additionally included.

[0035] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, or suppositories. Non-aqueous solutions and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin, glycerol, and gelatin.

[0036] The composition of the present invention can be administered orally or parenterally, and when administered parenterally, can be formulated in the form of a topical agent for skin; an injection; a transdermal agent; or a nasal inhalant according to a method known in the art.

[0037] In the case of the above injection, it must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Examples of suitable carriers for the injection include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, and thimerosal may be additionally included. In addition, the injection may in most cases additionally include isotonic agents such as sugars or sodium chloride.

[0038] For inhalation administration, the compounds used according to the present invention may conveniently be delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or another suitable gas. For pressurized aerosols, the dosage unit may be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

[0039] The composition of the present invention is administered in a pharmaceutically effective amount. A pharmaceutically effective amount refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dosage level can be determined based on the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concomitant drugs, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. That is, the total effective amount of the composition of the present invention can be administered to a patient as a single dose, or can be administered as a fractionated treatment protocol in which multiple doses are administered over a long period of time. It is important to consider all of the above factors and administer an amount that achieves the maximum effect with the minimum amount without causing side effects, and this can be easily determined by those skilled in the art.

[0040] The dosage of the pharmaceutical composition of the present invention varies depending on the patient's body weight, age, sex, health status, diet, administration time, administration method, excretion rate, and disease severity. The dosage may be increased or decreased depending on the route of administration, severity of obesity, sex, body weight, age, etc.

[0041] The composition of the present invention may be used alone or in combination with methods using chemotherapeutic and biological response modifiers.

[0042]

[0043] Another aspect of the present invention provides a method for preventing or treating a disease related to cognitive dysfunction or memory impairment, comprising the step of administering the pharmaceutical composition to a subject.

[0044] The above “cognitive dysfunction”, “memory impairment”, “prevention”, and “improvement” are as described above.

[0045]

[0046] Another aspect of the present invention provides a food composition for improving cognitive function or memory, comprising a perilla frutescensvar. acutakudo extract and a peppermint (Mentha piperita) extract.

[0047] The term "food" used in the present invention includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the conventional sense.

[0048] Since the food composition of the present invention can be consumed on a daily basis, it can be expected to have an effect of improving cognitive function or memory, and thus can be very usefully used for health promotion purposes.

[0049] The above functional health food is the same term as food for special health use (FoSHU), and refers to a food with high medical and healthcare effects that is processed to efficiently exhibit a bioregulatory function in addition to providing nutrition. Here, 'functionality' means regulating nutrients for the structure and function of the human body or obtaining a useful effect for health purposes such as physiological action. The food of the present invention can be manufactured by a method commonly used in the art, and during the manufacturing process, raw materials and ingredients commonly added in the art can be added. In addition, the formulation of the food can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms of formulations, and 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, and is highly portable, so the food of the present invention can be consumed as a supplement to enhance cognitive function or memory improvement.

[0050] The term "health food" refers to foods that have a more active health maintenance or promotion effect than regular foods, while "health supplement food" refers to foods intended for health supplementation. In some cases, the terms "health functional food," "health food," and "health supplement food" are used interchangeably.

[0051] Specifically, the health functional food means a food product manufactured by adding the compound of the present invention to food materials such as beverages, teas, spices, gums, and confectionery, or by manufacturing it in the form of encapsulation, powder, suspension, etc., and which has a specific health effect when consumed, but unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs made from food as a raw material.

[0052] The above food composition may additionally include a physiologically acceptable carrier. The type of the carrier is not particularly limited, and any carrier commonly used in the art may be used.

[0053] In addition, the food composition may include additional ingredients commonly used in food compositions to improve odor, taste, appearance, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and amino acids such as lysine, tryptophan, cysteine, and valine.

[0054] In addition, the food composition may include food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), sterilizers (bleaching powder and high-purity bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar colorants, etc.), color developers (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, sodium saccharin, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (glucose fillers), film-forming agents, gum-forming agents, foam suppressants, solvents, and improvers. The above additives can be selected according to the type of food and used in an appropriate amount.

[0055] As an example, the food composition of the present invention can be used as a health beverage composition, and in this case, it can contain various flavoring agents or natural carbohydrates as additional ingredients like a regular beverage. The above-mentioned natural carbohydrates can be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener can be a natural sweetener such as thaumatin and stevia extract; a synthetic sweetener such as saccharin and aspartame, etc. The proportion of the natural carbohydrate can be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g per 100 mL of the health beverage composition of the present invention.

[0056] In addition to the above, the health beverage composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in combination. The proportion of these additives is not particularly important, but is typically selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health beverage composition of the present invention.

[0057]

[0058] Another object of the present invention is to provide a feed composition for improving cognitive function or memory, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

[0059] The term "feed" as used in the present invention means any natural or artificial diet, meal, etc. or ingredients of said meal for eating, ingesting, and digesting by an animal or suitable therefor.

[0060] The type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include plant feed such as grains, roots, food processing by-products, algae, fiber, pharmaceutical by-products, oils, starches, meal, or grain by-products; and animal feed such as proteins, inorganic substances, oils, minerals, oils, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0061] A composition for improving cognitive function or memory, comprising a perilla frutescensvar. acutakudo extract and a peppermint (Mentha piperita) extract according to one embodiment of the present invention, has few side effects, is easy to access, is economical, and has an excellent effect of improving cognitive function, and thus can be usefully used for the prevention, improvement, or treatment of diseases related to cognitive function or memory impairment.

[0062] Figure 1 is a schematic diagram showing the overall experimental process according to one embodiment of the present invention.

[0063] Figure 2 shows the efficacy of improving cognitive function according to a Y-maze test when 10 mg / kg of a natural extract mixture consisting of chazzi and peppermint is orally administered in a cognitive function-impaired mouse model according to one embodiment of the present invention.

[0064] Figure 3 shows the cognitive function improvement efficacy according to a passive avoidance test when 10 mg / kg of a natural extract mixture consisting of chazzi and peppermint was orally administered in a cognitive function-impaired mouse model according to one embodiment of the present invention.

[0065] Figure 4 shows the efficacy of improving cognitive function according to a Y-maze test when 20 mg / kg of a natural extract mixture consisting of chazzi and peppermint is orally administered in a cognitive function-impaired mouse model according to one embodiment of the present invention.

[0066] Figure 5 shows the efficacy of improving cognitive function in a passive avoidance test when 20 mg / kg of a natural extract mixture consisting of chazzi and peppermint is orally administered in a cognitive function-impaired mouse model according to one embodiment of the present invention.

[0067] Figure 6 shows the cognitive function improvement efficacy according to the Y-maze test when 10 mg / ml of a natural extract mixture consisting of chazzi and peppermint was inhaled in a cognitive function impaired mouse model according to one embodiment of the present invention.

[0068] Figure 7 shows the neuroprotective efficacy of a natural extract mixture composed of tea tree and peppermint according to one embodiment of the present invention.

[0069] Hereinafter, the present invention will be described in more detail through examples. The following examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0070]

[0071] Before describing the examples, the overall experimental process for evaluating the cognitive function and memory improvement efficacy of the extract mixture of the present invention is shown in Figure 1.

[0072]

[0073] Example 1. Preparation of tea tree extract and peppermint extract

[0074] Perilla frutescensvar. acutakudo) or peppermint (Mentha piperita) was extracted with hot water at 80°C and then sterilized at 121°C for 20 minutes to prepare a hot water extract of Perilla frutescensvar. acutakudo or peppermint (Cha W, Pe W).

[0075]

[0076] Example 2. Creation of a sleep deprivation (SD) stress model

[0077] To induce cognitive impairment in mice, a sleep deprivation stress model was created. First, eight platforms were installed in the rat cage and four in the mouse cage, and water was filled to 1 cm below the platforms. When experimental animals were placed on the platforms, they would fall asleep or become drowsy. This principle was exploited: if their bodies touch the water, they would not be able to reach sleep. Therefore, rats or mice were placed in each cage and sleep was deprived continuously for approximately 72 hours, with the water changed every 24 hours.

[0078] For the sleep deprivation stress model produced in this way, when administered orally, different doses of each of the tea tree and peppermint extracts or the extract mixture were directly administered, and when administered by inhalation, a 5 cm × 5 cm gauze pad sufficiently absorbed with 5 ml of inhalant material including each of the tea tree and peppermint extracts or the extract mixture was attached to the top of the animal cage equipped with a filter, and then the mouse was placed on the sleep deprivation platform and the lid was closed to allow the smell to spread inside the platform.

[0079]

[0080] Example 3. Y-maze test

[0081] To determine whether a natural extract mixture was effective in mice with cognitive impairment, a Y-maze test was conducted. The Y-maze test, designed to assess the spontaneous alternation behavior of mice, was performed on an acrylic plate measuring 3 cm in width, 36 cm in length, and 15 cm in height. After designating each zone as A, B, and C, a mouse was carefully placed in one zone, and the mouse's movement path through the zone it entered was observed and recorded for 8 minutes. Afterwards, the number and order of entries into each zone were measured to assess spontaneous alternation. One point was awarded for sequential entries into three different zones, and no points were scored for non-consecutive entries. Alternation behavior (%) was calculated using the following formula.

[0082] [Calculation formula]

[0083]

[0084]

[0085] Example 4. Passive avoidance task

[0086] To determine whether a natural extract mixture was effective in mice with cognitive impairment, a passive avoidance test was conducted. The passive avoidance test assesses the short-term memory of mice for unpleasant stimuli. The test was conducted in a shuttle box consisting of two chambers (bright and dark) made of black polyvinyl plastic (20 × 20 × 20 cm in size) with a 2-mm grid on the floor, separated by a guillotine door. The bright chamber was equipped with a 50-W light bulb, and the dark chamber was connected to an electrical stimulator so that electricity could flow along the grid. The mouse was placed in the bright compartment and given a 20-second exploration period, after which the guillotine door was opened to allow entry into the dark compartment. After confirming that all four paws had entered, the door was closed, and a 0.5 mA electrical stimulus was applied for 3 seconds to initiate fear learning (acquisition trial).

[0087] Twenty-four hours later, the mice were placed back in the bright compartment, and after a 20-second exploration period, the guillotine door was opened to allow entry into the dark compartment. The time it took for all four paws to enter the dark compartment (latency time) was measured (retention trial). Mice that did not move to the dark compartment within 40 seconds after the guillotine door was opened were excluded from the experiment.

[0088]

[0089] Example 5. Measurement of cell viability

[0090] To confirm the neuroprotective efficacy of the natural extract mixture according to the present invention, cell viability was evaluated using the MTT assay.

[0091] First, HT22 cells, which are hippocampal cells, were seeded at 5 × 10 per well in a 96-well plate.3 After dividing the cells, they were cultured for 24 hours. After that, the cells were divided and pretreated for 2 hours with single natural extracts and natural extract mixtures dissolved in DPBS (Dulbecco's Phosphate-Buffered Saline). After pretreatment, glutamate (final concentration 10 mM) was added to each well and incubated for an additional 24 hours. After that, MTT reagent (final concentration 5 mg / mL, pH 7.4 dissolved in DPBS) was added to each well and the reaction was continued for a certain period of time. After MTT reagent treatment, 100 μL of DMSO was added to each well to dissolve the formed formazan crystals, and the reaction was continued at room temperature for a certain period of time, and the absorbance (OD) value was measured at 540 nm.

[0092]

[0093] Hereinafter, the effects of the present invention will be described through experimental examples. However, the effects of the present invention are not limited to the following experimental examples.

[0094]

[0095] Experimental Example 1. Confirmation of cognitive improvement efficacy when 10 mg / kg of a natural extract mixture is administered orally in a mouse model of cognitive impairment.

[0096] In order to confirm the cognitive function improvement efficacy of the natural extract mixture consisting of chasteberry and peppermint according to the present invention, chasteberry water extract and peppermint water extract were prepared in the same manner as in Example 1, and then 0.9% saline (VEH), chasteberry water extract 10 mg / kg (ChaW), peppermint water extract 10 mg / kg (PeW), and chasteberry water extract 5 mg / kg + peppermint water extract 5 mg / kg (ChaWPeW) were orally administered to the sleep-deprived mouse model of Example 2 at set times for 3 days to create a control group and an experimental group. Thereafter, the cognitive function improvement efficacy of the natural extract mixture was confirmed through a cognitive ability evaluation for the control group and the experimental group, and the results are shown in FIGS. 2 and 3.

[0097] Looking at Figure 2, in the case of the 0.9% saline administration group (VEH), the behavioral change ability decreased compared to the control group (CTL) when the Y-maze test was performed. However, when orally or inhaledly administered 10 mg / kg of chaste tree water extract (ChaW) or 10 mg / kg of peppermint water extract (PeW), the behavioral change ability increased compared to VEH, and in the case of the group administered 5 mg / kg of chaste tree water extract + 5 mg / kg of peppermint water extract (ChaWPeW), the behavioral change ability was confirmed to have recovered to a level close to the control group compared to the single material administration group.

[0098] Looking at Figure 3, in the case of the 0.9% saline administration group (VEH), the latency decreased in the retention trial when the passive avoidance test was performed. However, when orally administered 10 mg / kg of perilla water extract (ChaW) or 10 mg / kg of peppermint water extract (PeW), the latency in the retention trial was equivalent to or increased compared to VEH, and in the case of the group administered 5 mg / kg of perilla water extract + 5 mg / kg of peppermint water extract (ChaWPeW), the latency in the retention trial significantly increased compared to the single material administration group, confirming that cognitive ability was restored to a level close to the control group.

[0099]

[0100] Experimental Example 2. Confirmation of cognitive improvement efficacy when 20 mg / kg of a natural extract mixture is administered orally in a mouse model of cognitive impairment.

[0101] In order to confirm the cognitive function improvement efficacy of the natural extract mixture comprising chajiki and peppermint according to the present invention, a control group and an experimental group were prepared using the same method as Experimental Example 1, but with the amounts of chajiki extract and peppermint extract doubled. Thereafter, the cognitive function improvement efficacy of the natural extract mixture was confirmed through cognitive ability evaluation for the control and experimental groups, and the results are shown in FIGS. 4 and 5.

[0102] Looking at Figure 4, in the case of the 0.9% saline administration group (VEH), the behavioral change ability decreased when the Y-maze test was performed. However, when orally administered 20 mg / kg of the water extract of Chajigi (ChaW) or 20 mg / kg of the water extract of peppermint (PeW), the behavioral change ability increased compared to VEH, and in the case of the group administered 10 mg / kg of the water extract of Chajigi + 10 mg / kg of the water extract of peppermint (ChaWPeW), the behavioral change ability was confirmed to be higher than the control group, confirming that cognitive function was completely recovered.

[0103] Looking at Figure 5, in the case of the 0.9% saline administration group (VEH), the latency slightly increased in the retention trial when the passive avoidance test was performed. However, in the case of the 20 mg / kg administration group of the perilla water extract (ChaW) and the 20 mg / kg administration group of the peppermint water extract (PeW), the latency in the retention trial increased significantly compared to VEH, and in the case of the 10 mg / kg administration group of the perilla water extract + 10 mg / kg administration group of the peppermint water extract (ChaWPeW), the latency in the retention trial increased further compared to the single material administration group, confirming that the cognitive ability was restored to a level similar to the control group.

[0104]

[0105] Experimental Example 3. Confirmation of cognitive improvement efficacy when inhaled with 10 mg / ml of a natural extract mixture in a mouse model of cognitive impairment.

[0106] In order to confirm the cognitive function improvement efficacy when a natural extract mixture consisting of chazuki and peppermint according to the present invention is administered by inhalation, chazuki water extract and peppermint water extract were prepared in the same manner as in Example 1, and then 0.9% saline (VEH), chazuki water extract 10 mg / ml (Cha W), peppermint water extract 10 mg / ml (Pe W), and chazuki water extract 5 mg / ml + peppermint water extract 5 mg / ml (Cha W Pe W) were administered by inhalation to a sleep-deprived mouse model of Example 2 at set times for 3 days to create a control group and an experimental group. Thereafter, the cognitive function improvement efficacy of the natural extract mixture was confirmed through a cognitive ability evaluation for the control group and the experimental group, and the results are shown in Fig. 6.

[0107] Looking at Figure 6, in the case of the 0.9% saline administration group (VEH), the behavioral change ability decreased compared to the control group (CTL) when the Y-maze test was performed. However, when inhaled, 10 mg / ml of citronella tea tree water extract (ChaW) or 10 mg / ml of peppermint water extract (PeW) increased the behavioral change ability compared to VEH, and in the case of the group administered 5 mg / ml of citronella tea tree water extract + 5 mg / ml of peppermint water extract (ChaWPeW), the behavioral change ability was confirmed to have recovered to a level close to the control group compared to the single material administration group.

[0108]

[0109] Experimental Example 4. Confirmation of the neuroprotective efficacy of a natural extract mixture composed of chaga and peppermint.

[0110] In order to confirm the neuroprotective efficacy of the natural extract mixture composed of tea tree and peppermint according to the present invention, cell viability was evaluated through an MTT assay in the same manner as in Example 5, and the results are shown in Fig. 7.

[0111] Looking at Figure 7, in the 0.9% saline administration group (VEH), the cell viability decreased by nearly half compared to the untreated control group. However, in the group administered with the water extract of Chajigi (ChaW) and the group administered with the water extract of peppermint (PeW), the cell viability increased compared to VEH, and in the group administered with the water extract of Chajigi + the water extract of peppermint (PeW), it was confirmed that the cell viability recovered to a level close to the control group compared to the single material administration group.

[0112]

[0113] Based on the above results, it was confirmed that when a mixture of chazzi and peppermint extracts was administered orally or by inhalation, cognitive function and memory improvement efficacy were significantly improved compared to the single-material administration group, and the cell protection effect was also significantly more effective than the single-material administration group, clearly confirming that the natural extract mixture showed a synergistic effect compared to the single-material administration group.

[0114]

[0115] While the present invention has been described with reference to one embodiment, this is merely exemplary, and those skilled in the art will understand that various modifications and equivalent alternative embodiments are possible. Therefore, the true scope of technical protection of the present invention should be determined by the technical spirit of the appended claims.

[0116] A composition for improving cognitive function or memory, comprising a perilla frutescensvar. acutakudo extract and a peppermint (Mentha piperita) extract according to one embodiment of the present invention, has excellent effects in improving cognitive function, is based on natural materials, has few side effects, is easily accessible, and is economical, and thus can be usefully used in the prevention, improvement, or treatment of diseases related to cognitive function or memory impairment, and thus has industrial applicability.

Claims

1. A composition for improving cognitive function or memory, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

2. In paragraph 1, A composition for improving cognitive function or memory, wherein the above extraction is performed using water, an organic solvent, or a mixture thereof as an extraction solvent.

3. In paragraph 1, The composition is a composition for improving cognitive function or memory, characterized in that the tea extract and peppermint extract are mixed in a weight ratio of 1:0.5 to 1:

4.

4. A pharmaceutical composition for preventing or treating diseases related to cognitive dysfunction or memory impairment, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

5. In paragraph 4, A pharmaceutical composition, wherein the above cognitive dysfunction or memory impairment-related disease is a degenerative brain nervous system disease.

6. In paragraph 4, A pharmaceutical composition, wherein the above cognitive dysfunction or memory impairment-related disease is mild cognitive impairment (MCI), Alzheimer's disease, or Parkinson's disease.

7. A method for preventing or treating a disease related to cognitive dysfunction or memory impairment, comprising a step of administering the pharmaceutical composition of paragraph 4 to a subject.

8. In paragraph 7, A method for preventing or treating a disease related to cognitive dysfunction or memory impairment, characterized in that the above administration is oral or inhaled.

9. In paragraph 7, A method for preventing or treating a disease related to cognitive dysfunction or memory impairment, wherein the disease related to cognitive dysfunction or memory impairment is a degenerative brain nervous system disease.

10. In paragraph 7, A method for preventing or treating a disease related to cognitive dysfunction or memory impairment, wherein the disease related to cognitive dysfunction or memory impairment is mild cognitive impairment (MCI), Alzheimer's disease, or Parkinson's disease.

11. A food composition for improving cognitive function or memory, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

12. A feed composition for improving cognitive function or memory, comprising an extract of Perilla frutescensvar. acutakudo and an extract of peppermint (Mentha piperita).

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