KR100564904B1 - Composition for preventing or treating dementia and improving cognitive function comprising herbal extract as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or a functional food composition for preventing or treating dementia and improving cognitive function, which includes licorice, earl pear, donkey, bark skin, sanjoin, baekbaekpi, changchangpo, horse riding, ointment, and gardenia extract as active ingredients. The extract has an inhibitory activity of inhibiting acetylcholinesterase or beta amyloid aggregation, and in particular, licorice, earl nectar, bark skin, horse riding and oak extract have excellent neuronal cell protection. By the above efficacy, the extracts may be usefully used in the composition for preventing or treating dementia and improving cognitive function. According to the animal efficacy test for licorice and earl pear, the licorice and earl medicinal herb extract shows excellent efficacy of improving the learning and spatial perception ability of the dementia animal group induced by beta amyloid to be close to the normal animal group.

Since the composition of the present invention contains the herbal extract as an active ingredient, the side effects on the human body are extremely less than that of the chemical synthetic medicine.

Dementia, prevention, treatment, cognition, acetylcholinesterase, beta amyloid, herbal extract, composition

Description

Composition for Preventing or Treating Dementia and Improving Cognitive Function Comprising Oriental Medicine Extracts as Active Ingredient}

The present invention relates to a composition for preventing or treating dementia and improving cognitive function, and more particularly, to a pharmaceutical composition for preventing or treating dementia and improving cognitive function and a functional food composition comprising an herbal extract as an active ingredient. .

As it became clear that the 21st century would become an aging society, the era of human beings seeking a high quality of life by living long and healthy without disease has arrived. As the life expectancy increases due to the development of science, the proportion of dementia patients with neurodegenerative diseases is increasing rapidly. Dementia is a condition that causes a significant drop in quality of life, both personally and socially, and makes you and those around you unhappy. It is the fourth leading cause of death for the elderly, following cancer, heart disease and stroke. Dementia can occur in a number of ways, but first, it is highly related to a decrease in the concentration of a neurotransmitter called acetylcholine in the brain's hippocampus.

Acetylcholine, which has a quaternary amine structure, is a neurotransmitter and acetylcholine esterase acts to hydrolyze acetylcholine into choline. In patients with dementia, the concentration of acetylcholine, a neurotransmitter, is reduced, and the inhibition of acetylcholinesterase increases the concentration of acetylcholine in the brain, thereby improving symptoms of dementia patients. Drugs that have been developed and used in the treatment of dementia with FDA approval are tacrine, donepezil, galanthamine, and rivastigmine, all of which are acetylcholinesterase inhibitors.

Second, it modulates genetic factors related to the production, progression, accumulation and deposition of neuronal beta amyloid to slow disease progression and to selectively remove extracellular concentrations and beta amyloid deposits in the brain. To find out. Beta-amyloid peptide aggregation is a fundamental cause of dementia, and studies are being actively conducted to suppress it.

Third, one can consider secondary ways to prevent disease by using estrogens, antioxidants, free radical scavengers and anti-inflammatory agents that can delay the onset of the dementia or slow or stop progression.

Fourth, it is a method of preventing the progressive and irreversible degeneration of synapses and neurons.

The development of treatment for dementia can reduce the difficulty of patients suffering from dementia and contribute to improving the quality of life of humans. Therefore, it is very important to develop a treatment for Alzheimer's disease with excellent physiological activity. However, the development of new drugs for dementia is too long, the cost of research is huge, and the risk of failure is very high. The fact that Korea has traditionally used herbal medicines that have been recognized by the private and clinical sectors has been a significant advantage as it has the possibility of being applied to bio foods during the new drug development and new drug development process.

If a drug can be searched for a drug that has the function of preventing or treating dementia by inhibiting acetylcholinesterase or a dementia-inducing protein, beta-amyloid aggregation, a mechanism of action that can prevent or treat dementia, and apply it to pharmaceutical compositions or foods for a short time. It is expected to be an important base technology for new drug development if high value can be created within it, and further, the active fractions can be separated and purified to identify the structure of the active substance.

The present inventors have diligently researched to find herbal medicines with excellent physiological effects in improving dementia and cognitive function. As a result, the inventors have found that licorice, earl pear, donkey, bark, sanjoin, baekbaekpi, spearmint, horse riding, ointment, and gardenia extracts inhibit acetylcholinesterase or Beta-amyloid coagulation inhibitory activity, licorice, earl, bark, horseback riding and ointment extracts were found to have excellent neuronal cell protection, licorice and earl extract were enhanced beta amyloid-induced learning and spatial perception The present invention has been completed by confirming that it exhibits excellent efficacy of improving the ability close to normal animals.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition comprising: (a) a pharmaceutically effective amount of one or more herbal extracts of licorice, earl, donkey, bark, sanjoin, lettuce, spearmint, horse riding, ointment and gardenia extract; And (b) to provide a pharmaceutical composition for preventing or treating dementia comprising a pharmaceutically acceptable carrier.

It is another object of the present invention to provide a functional food composition for preventing or treating dementia comprising as an active ingredient one or more herbal extracts of licorice, earl pear, Angelica, bark, sanjoin, lettuce, spearmint, horseback riding, oak, and gardenia extract. .

Another object of the present invention is (a) a pharmaceutically effective amount of one or more herbal extracts of licorice, earl, donkey, bark, sanjoin, lettuce, spearmint, horseback riding, oak and gardenia extracts; And (b) to provide a pharmaceutical composition for improving cognitive function comprising a pharmaceutically acceptable carrier.

It is another object of the present invention to provide a functional food composition for improving cognitive function, which comprises one or more herbal extracts of licorice, earl pear, Angelica, bark, sanjoin, baekbaek, changchang, horseback riding, oak, and gardenia extract as active ingredients.

Other objects and advantages of the present invention will become apparent from the following examples and claims.

According to one aspect of the present invention, the present invention provides a pharmaceutical composition comprising: (a) a pharmaceutically effective amount of one or more herbal extracts of licorice, earl pear, donkey, bark skin, sanjoin, lettuce, spearmint, horse riding, oak and gardenia extract; And (b) provides a pharmaceutical composition for preventing or treating dementia comprising a pharmaceutically acceptable carrier.

According to another aspect of the present invention, the present invention is a functional food for preventing or treating dementia comprising as an active ingredient an extract of at least one herbal extract of licorice, earl pear, donkey, bark, sanjoin, lettuce, spearmint, horse riding, oak and gardenia extract To provide a composition.

The present invention is characterized by comprising a herbal extract as an active ingredient.

Licorice, earl medicinal herb, donkey, bark skin, Sanjoin, Sangbaekpi, Seokchangpo, horse riding, oak and gardenia, which are used in the present invention, have been widely used as herbal medicines.

The herbal extracts may be prepared by various extraction solvents such as (a) water, (b) anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) the lower alcohols and water Mixed solvent, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3-butylene glycol and (h) butyl acetate can be obtained as an extraction solvent. Preferably, the extract of the present invention is obtained using a hydrous lower alcohol, more preferably methanol, most preferably 70% methanol. On the other hand, it will be apparent to those skilled in the art that the extract of the present invention can be obtained not only with the aforementioned extraction solvent but also with other extraction solvents.

In addition, the extract of the present invention includes not only the extract by the above-mentioned extraction solvent, but also an extract that has undergone a conventional purification process. Obtained by various additional purification methods, such as, for example, separation using ultrafiltration membranes having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). The active fraction is also included in the extract of the present invention.

Herbal extracts of 10 species of the present invention (Licorice, Earl of Pears, Angelica, Bark, Sanjoin, Morus bark, Seokpo, Horseback Riding, Suppository and Gardenia) have the activity of inhibiting acetylcholinesterase or inhibiting beta amyloid aggregation, thereby preventing or treating dementia. It is effective. In addition, licorice, earl nectar, bark skin, horse riding and oak extracts have excellent neuronal protection and are more useful for the prevention or treatment of dementia. According to the animal efficacy test for licorice and earl pear, which can be used as a food material, the two herbal extracts are excellent for improving the learning and spatial perception ability of the dementia animal group induced by beta amyloid close to the normal animal group. It shows efficacy.

According to a preferred embodiment of the present invention, the composition of the present invention preferably comprises as an active ingredient extract of at least one of the licorice, earl pear, Angelica, Bark skin, Sanjoin, Sangbaekpi, Sukchangpo, horse riding, oak and gardenia extract More preferably, one or more herbal extracts of licorice, earl berry, bark, horse riding and gardenia extract, most preferably licorice and earl berry extract as active ingredients.

As used herein, the term "prevention" means that it has not been diagnosed as having a disease or condition but inhibits the occurrence of the disease or condition in an animal that tends to be prone to such disease or condition. As used herein, the term “treatment” means (a) inhibiting the development of a disease or condition; (b) alleviation of the disease or condition; And (c) elimination of the disease or condition.

Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation of carbohydrate compounds, such as lactose, amylose, dextrose, sucrose, sorbitol, mannitol, starch, cellulose, and the like. ), Acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, salt solution, alcohol, gum arabic, vegetable oil (e.g. corn oil, cotton Seed oil, soy milk, olive oil, coconut oil), polyethylene glycol, methyl cellulose, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil, and the like. The pharmaceutical composition of the present invention further includes, but is not limited to, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like, in addition to the above components.

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, or the like.

Suitable dosages of the pharmaceutical compositions of the invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to reaction, Usually a skilled practitioner can easily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the invention, suitable dosages are from 0.1 mg / kg to 600 mg / kg once daily on an adult basis.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporating 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 an aqueous medium, or may be in the form of extracts, powders, granules, tablets or capsules, and may further include a dispersant or stabilizer.

On the other hand, the functional food composition of the present invention includes components that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, in the case of the production of a drinking agent, in addition to the silk peptide of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, and the like may be further included. Given the ease of access to food, the food of the present invention is very useful for treating or preventing dementia.

According to a preferred embodiment of the present invention, the content of the herbal composition in the functional food composition of the present invention is 0.001% to 50% by weight.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition comprising: (a) a pharmaceutically effective amount of one or more herbal extracts of licorice, earl pear, donkey, bark, sanjoin, lettuce, spearmint, horse riding, oak and gardenia extracts; And (b) provides a pharmaceutical composition for improving cognitive function comprising a pharmaceutically acceptable carrier.

According to another aspect of the present invention, the present invention (a) for improving the cognitive function comprising one or more herbal extracts of licorice, earl pear, donkey, bark skin, sanjoin, baekbaekpi, Seokchangpo, horse riding, oak and gardenia extract as an active ingredient Provides a functional food composition.

The herbal extract of the present invention shows excellent efficacy in improving cognitive function.

As used herein, the term "improving cognitive function" includes improving learning and / or spatial perception. In the present invention, "improvement of learning ability and / or spatial perception ability" is interpreted to include the meaning of improvement of memory ability.

Acetylcholine is a neurotransmitter that is projected from the basal ganglia of the brain to the cerebral cortex and hippocampus and is very important for normal knowledge function (Richter et. Al., Life Sci. 19; 26 (20): 1683-9 (1980) ). In particular, it is known that learning and memory can be altered by drugs acting on the acetylcholine system.

The herbal extract of the present invention prevents impairment of cognitive function, treats impaired cognitive function, prevents impairment of cognitive function, and improves cognitive function through inhibition of acetylcholinesterase or inhibition of beta amyloid aggregation and neuronal protective effect. It works. According to the animal efficacy test for licorice and earl pear, the two herbal extracts show excellent efficacy of improving the learning and spatial perception ability of the dementia animal group induced by beta amyloid close to the normal animal group.

Suitable dosages of the pharmaceutical composition for improving cognitive function are from 0.1 mg / kg to 600 mg / kg once daily on an adult basis. According to a preferred embodiment of the present invention, the content of the herbal composition in the food composition for improving cognitive function of the present invention is 0.001% to 60% by weight.

In addition, in common with the composition for preventing or treating dementia mentioned above, the description is abbreviate | omitted in order to avoid excessive complexity of this specification by a duplicate base material.

Since the composition of the present invention not only exhibits the above-mentioned efficacy, but also contains the herbal extract as an active ingredient, the side effects on the human body are extremely less than that of the chemical synthetic medicine.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it is 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. Will be self-evident.

Example

Example 1 Preparation of Herbal Extracts

The herbal samples used in the experiments were purchased from Kyungdong market and used in the experiment after accurate evaluation by experts. 100 g of the dried herbal sample was placed in 70% methanol 1 ° and extracted under reflux for 3 hours, and filtered using a filter paper. The filtrate was concentrated in a rotary evaporator and immediately lyophilized.

Experimental Example 1: Determination of acetylcholinesterase inhibition and beta amyloid aggregation inhibition

Experimental Example 1-1 Reagent

Acetylcholinesterase (type VS), acetylcholine iodide, 5,5-dithio-bis (2-nitrobenzoic acid), neostigmine bromide used to measure the activity of acetylcholinesterase (neostigmine bromide) was a product of Sigma (United States of America).

Beta amyloid 1-42, used to measure the degree of aggregation of beta amyloid, is a product of American Peptide Co. of the United States, thioflavin T, and phosphate buffer solution tablet ( Phosphate buffered saline tablets and dimethyl sulfoxide (DMSO 99.5%) were used by Sigma, USA. As other reagents, Duksan Science's express or first-class reagents were used.

Experimental Example 1-2: Method for Measuring Acetylcholinesterase Inhibition Activity

Acetylcholinesterase inhibitory activity measurement was performed by modifying the Ellman method. The enzyme was aliquoted and stored at -80 ° C, the final concentration was used at a concentration of 0.03 units, and the substrate was used with 1 mM acetylcholine iodide dissolved in 0.1 M sodium phosphate buffer (pH 8.0). The color development reagent was prepared by dissolving 39.6 mg of 5,5-dithio-bis (2-nitrobenzoic acid) and 15 mg of sodium bicarbonate in 10 ml of 0.1 M sodium phosphate buffer (pH 7.0). The enzymatic reaction was developed as follows. 20 μl of herbal extract (prepared to 1 mg / ml in final reaction solution), 200 μl of 10 mM DTNB solution and 100 μl of enzyme (0.03 U) in 2 ml of 0.1 M sodium phosphate buffer (pH 8.0) After the addition and pre-incubation for 10 minutes at 37 ℃, 200 μl of the substrate was added to react for 3 minutes and the absorbance (412 nm) was measured.

Acetylcholinesterase inhibitory activity was calculated by the following equation.

% Inhibition = [1- (test O.D.-test O.D.) / (Control O.D.-control O.D.)] X100

Experimental Example 1-3: Method for measuring the degree of aggregation of beta amyloid 1-42

The degree of aggregation of beta amyloid was measured by thioflavin analysis using a fluorescence spectrometer. The peptide was stored at minus 80 ℃, the final concentration used was 25 μM, and the herbal extract to be measured was dissolved in 99.5% DMSO. 5 μM of thioflavine T used as a fluorescent material was prepared by dissolving in 0.01 M sodium phosphate buffer solution. The aggregation reaction was developed as follows. To 92.5 μl of 0.01 M sodium phosphate buffer solution, 5 μl of the herbal extract made at a constant concentration was added (prepared to 1 mg / ml), and 2.5 μl of 5 μM beta amyloid 1-42 was added. Time aggregation was performed, and 2000 µl of 5 µM thioflavin T was added, and the intensity was measured using a fluorescence spectrometer (RF-5300PC, Shimadzu Corp.). The wavelength value of the fluorescence spectrometer used at this time was Ex = 446 nm (slit width = 5 nm) and Em = 490 nm (slit width = 10 nm).

Experimental Example 1-4: Measurement Result

70% methanol extracts of 40 herbal drugs obtained in Example 1 were tested for their ability to inhibit acetylcholinesterase and to inhibit the aggregation of beta amyloid peptide, which is a dementia-inducing protein. The results are shown in Table 1 below.

Test Results of Acetylcholinesterase Inhibition and Aβ Aggregation Inhibition of Herbal Extracts No. Herbal medicine Latin herbal medicine AChE inhibition (%, 1 mg / ml) Aβ aggregation inhibition (%, 1 mg / ml) NP01 licorice Glycyrrhizae radix 6.4 86.4 NP02 Earl Paeoniae radix 69.3 85.8 NP03 Wolfberry Lycii Fructus 2.82 31.6 NP04 health Zingiberis Rhizoma 13.96 39 NP05 Donkey Angeilcae Gigantis Radix 27.40 72.8 NP06 contrast Zizyphi Fructus 0 0 NP07 Ginseng Codonopsis Pilosulae Radix 0 40 NP08 Neck skin Moutan Radicis Cortex 91.47 29.8 NP09 elecampane Saussureae Radix 53.29 48.9 NP10 Double Poria 13.67 0 NP11 Whiteness Atractylodis Rhizoma alba 12.95 10.6 NP12 Sanjoin Zizyphy Spinosi Semen 77.78 18.5 NP13 White skin Mori Cortex Radicis 77.49 45.9 NP14 Seokchangpo Acori Graminei Rhizoma 43.10 76.5 NP15 Shiho Bupleuri radix 12.24 0 NP16 riding Cimicifugae Rhizoma 73.11 44.4 NP17 Paper Polygalae radix 8.28 51.5 NP18 Oak Linderae Radix 76.65 71.6 NP19 Dragon meat Longanae arillus 0 0

No. Herbal medicine Latin herbal medicine AChE inhibition (%, 1 mg / ml) Aβ aggregation inhibition (%, 1 mg / ml) NP20 Jimo Anemarrhenae Rhizoma 9.21 43.1 NP21 Gardenia Gardeniae fructus 67.83 73.9 NP22 Shouao Polygoni Multiflori Radix 5.06 0 NP23 Astragalus Astragali radix 3.16 3.5 NP24 Go Terminaliae Fructus 21.31 10.4 NP25 curcuma Curcumae Longae Rhizoma 29.53 36.5 NP26 Gold coins Lonicerae Flos 22.18 37.2 NP27 Construction Amomi semen 14.31 0 NP28 male Arisaematis Rhizoma 55.56 23.5 NP29 Misam Ginseng radix 5.62 2.1 NP30 Banha (legal) Pinelliae Rhizoma 16.24 2.9 NP31 White sleep Bombycis corpus 37.00 28.1 NP32 Medicine Dioscoreae Radix 12.33 21.5 NP33 Windproof Ledebouriellae radix 0 0 NP34 Ginseng Ginseng radix 3.75 5.4 NP35 dermis Citri rericarpium 41.87 48.5 NP36 Kill Bambisae Caulis In Taeniam 5.22 36.7 NP37 Bad Polyporus 21.71 35.2 NP38 Cheonma Gastrodiae Rhizoma 47.46 28.9 NP39 Red ginseng Ginseng radix 27.38 22.8 NP40 Street view Platycodi Radix 0 29.1

Through this test, 10 kinds of herbal medicines with more than 70% activity at the concentration of 1 mg / ml of herbal extracts were selected: licorice, earl pear, donkey, bark, sanjoin, lettuce, changchang, horse riding, oak and gardenia.

Experimental Example 2: Dementia cell model fabrication and neuronal cell damage suppression

Experimental Example 2-1: Reagent

Amyloid-beta 1-42 peptide (amyloid-beta 1-42 peptide, Aβ _1-42 peptide) was purchased from American peptide company, Inc. MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide] reagent was purchased from Sigma, and general reagents including acetic acid were purchased from Duksan Science.

Experimental Example 2-2: Cell Culture and MTT Analysis

Human neurons, IMR-32 cells (ATCC, US), were treated with 10% heat-inactivated fetal calf serum, Gibco BRL, Gaithersburg, Md., USA, penicillin (100 units / ml) and streptomycin. Incubated in EMEM (ATCC, USA) medium to which (100 g / mL) was added at 37 ° C. and 5% CO ₂ until logarithmic growth. Cultured IMR-32 cells were placed in a 96-well plate at a concentration of 9x10 ⁴ cells / cm ² , stabilized for 2 hours, and then beta amyloid alone (control), beta amyloid (final concentration 20 μM / 0.1% acetic acid) and herbal extracts. These cells were divided into groups treated with 0.1 mg / ml, respectively, and cultured at 37 ° C. and 5% CO ₂ for 48 hours. MTT reagent (5 mg / ml) was added at 50 µl / well for 4 hours, and 100 µl / well of DMSO was added, followed by 570 nm using an ELISA plate reader (Bio-Tek Instruments. Inc., Vermont, USA). Absorbance was measured at. The test was calibrated and the same test was performed three or more times in three wells.

Neuroprotective capacity for neurons was calculated by the following equation (2).

Cell viability (%) = (test O.D.-test background O.D./control O.D.-control background O.D.) x100

Experimental Example 2-3: Results of Measurement of Neuronal Protective Activity of Herbal Extracts against Dementia Cell Models

Ten medicinal herbs with acetylcholinesterase inhibitory activity or beta amyloid peptide aggregation inhibitory ability of 70% or more were selected and tested for neuronal cell protection against dementia cell models. The results are shown in Table 2 below.

Neuroprotective Activity of Natural Extracts Against Neuronal Cell Line, IMR-32 by MTT Assay No. Herbal medicine Latin herbal medicine Cell viability (0.1mg / ml) - Control Aβ1-42 20 uM 49.1% NP01 licorice Glycyrrhizae radix 90.4% NP02 Earl Paeoniae radix 90.1% NP05 Donkey Angeilcae Gigantis Radix 70.6% NP08 Neck skin Moutan Radicis Cortex 90.0% NP12 Sanjoin Zizyphy Spinosi Semen 72.1% NP13 White skin Mori Cortex Radicis 67.4% NP14 Seokchangpo Acori Graminei Rhizoma 70.4% NP16 riding Cimicifugae Rhizoma 80.1% NP18 Oak Linderae Radix 90.4% NP21 Gardenia Gardeniae fructus 68.0%

When the beta amyloid 1-42 was treated with the neuronal cell line IMR-32, the cell viability was 49.1% and about 50% of the neuronal cell inhibition was confirmed as a dementia cell model. The herbal extracts with cell viability of more than 80% were found in five categories: licorice, earl nectar, bark skin, horse riding and oak. Two herbicides were tested for dementia animal models.

Experimental Example 3: Dementia Animal Model

Experimental Example 3-1: Animals and Breeding Conditions

The male Black6 mouse was purchased from Korea Bio Link Co., Ltd. (Korea) for 4 weeks and was adapted to the environment of the nursery for 2 weeks and then used for the experiment. At the start of the experiment, they were 6 weeks old and the body weights ranged from 17-21 g. Breeding in plastic cages, gave enough water and feed. The nursery environment was subjected to a dark / light cycle of 12 hours and to maintain a temperature of 23 ± 1 ° C.

Experimental Example 3-2: Sample Preparation and Administration

Dementia was induced with beta amyloid (Aβ _1-42 ). Beta amyloid peptide was dissolved in 1% acetic acid so that the final concentration was 820 pico-mole, and 5 μl per animal was intracerebroventricular injection, and the control group was injected with 1% acetic acid as a solvent.

Licorice (or Earl) extract was dissolved in water and administered orally once a day. One dose was prepared to be 150 mg / kg. The dose was adjusted not to exceed 200 μl per day. Water was administered to the control group and beta amyloid alone treatment group.

Experimental Example 4: Effect of herbal extracts on animal behavior of dementia animal model (Y-maze test)

Experimental Example 4-1: Y-maze method

The instrument used for the Y-maze test consists of three branches, each arm is 34 cm long, 5 cm wide, 10 cm wide and 10 cm high, and the angle of contact between the arms is 120 °. It was. All the experiments were made of black and used with black tape to prevent light from leaking in. After setting each branch to A, B, C, the rat was carefully placed on one branch and allowed to move freely for 5 minutes, and then the branch containing the rat was recorded. Only when the tail is completely entered, it is also recorded when the branch went back. One point (actual alteration) was given to three different branches. Alteration behavior is defined as entering all three arms and is calculated by the following equation (3).

 % Change behavior = actual alteration / maxium alteration x 100.

(Maximum change: Total entry-2)

Experimental Example 4-2: Y-maze test results of herbal extracts for dementia animal model

Extracts of Licorice (NP1) and Earl (NP2) were subjected to a Y-maze test on animal models of dementia. The sample was treated by the method of Experimental Example 3-2, five black papers were used, and the results are shown in Table 3.

Aβ _1-42 Effect of NP1 and NP2 on Alteration Behavior in Y-maze of Rats Treated with group   Alteration Behavior (%) mean ± S.E.M   7PI ⁺ 15PI ⁺ Control  73.8 ± 7.19  73.8 ± 7.19 Ab _1-42 55.6 ± 7.00 ^## 48.2 ± 10.44 ^## NP1 64.8 ± 4.21 ^*  50.8 ± 7.44 NP2  55.8 ± 4.35  64.0 ± 3.90

+: PI = post injection

##: P <0.01, compared with control

*: P <0.05, compared with Aβ1-42 test group

7 days after 1% acetic acid was administered as icv (intracerebroventricular) (7PI), the average behavioral power of the control group was 73.8%, whereas beta amyloid group after 5 days with icv averaged 55.6% (p) <0.01) It was found that the dementia animal model was produced, and the behavioral power of the NP1 group was 64.8%, which was significantly increased (p <0.05) in the herbal extract group compared with the dementia group. Showed a tendency to increase, with an average of 55.8%.

After 15 days (15PI), the behavior of the control group was 73.8% on average, whereas the behavior of the beta amyloid group was significantly lower (p <0.01), indicating that a dementia animal model was produced. In the herbal extract-administered group, the behavioral power of the NP1-administered group was 50.8% on average, and the behavioral power of the NP2-administered group was 64.0% on average.

Experimental Example 5: Effect of herbal extracts on animal behavior of dementia animal model (manual evasion test)

Experimental Example 5-1: Passive avoidance test (PAT) method

The instrument used in the experiment consisted of a dark room and a light room. The floor was a grid floor through which current could flow, and a slightly larger hole than the rat was drilled between the two rooms to allow free traffic. The rats were placed in a clear room and left for free for one minute without lighting, and then illuminated for two minutes. At this time, the mouse does not like the bright stay in the dark room. Two minutes later, when the mouse enters the dark room, it closes the hole between the two rooms, gives an electric shock for two seconds, and opens the hole. At this time, the mouse opens and opens into the room. Two hours after the mice were transferred to cages, two PAT records were measured. Darkroom Enterance Latency (DEL) is the time the rat first enters the darkroom, and DSL (Darkroom Stay Latency) is the time to stay in the darkroom for 90 seconds. When measuring the PAT record, the lights were lit in the same room as the electric shock, but no current flowed.

Experimental Example 5-2: Manual evacuation test results of herbal extracts for animal models of dementia

The licorice (NP1) and Earl (NP2) extracts were subjected to passive avoidance tests on animal models of dementia. The sample was treated by the method of Experimental Example 3-2, five black papers were used, and the results are shown in Table 4.

Aβ _1-42 Effect of NP1 and NP2 on Dark Entry Time and Dark Stay Time in Passive Evasion Experiments (PAT) of Rats Treated with Group  Darkroom Entry Time (DEL)  (Sec) Average ± S.E.M Dark Room Retention Time (DSL) (Sec) Average ± S.E.M 7PI ⁺  15PI ⁺ 7PI ⁺ 15PI ⁺ Control   79.60 ± 8.21 79.60 ± 8.21   1.20 ± 1.17   1.20 ± 1.17 Ab _1-42 59.80 ± 6.94 ^# 22.60 ± 2.82 ^### 17.40 ± 3.32 ^### 6.20 ± 1.47 ^### NP1 85.20 ± 4.53 ^*** 91.40 ± 7.2 ^*** 4.60 ± 2.87 ^***   2.60 ± 3.88 NP2 84.00 ± 3.94 ^***   81.20 ± 16.13 7.60 ± 3.83 ^**   6.40 ± 1.20

+: PI = post injection

#: P <0.05, ###: P <0.001, compared to control (DEL)

***: P <0.05, compared with Aβ1-42 treatment group (DEL)

###: P <0.001, compared with control (DSL)

**: P <0.01, ***: P <0.001, compared with Aβ1-42 treatment group (DSL)

7 days after 1% acetic acid was administered with icv, the mean time to enter the dark room was 79.6 seconds in the passive evacuation ability of the control group, while beta amyloid group after 5 days with icv was significantly averaged 59.8 seconds (p < 0.05) was decreased, and the average time to stay in the dark room was 1.2 seconds in the control group, while the beta amyloid group was 17.4 seconds in average (p <0.001), indicating that a dementia animal model was produced. Compared to the dementia animals, the average drug extraction time of the NP1 group was 85.2 seconds in the dark room and 4.6 seconds in the dark room, showing significant significance (p <0.001) compared to the dementia animals. The average NP2 group was 84 seconds and 7.6 seconds, which also showed significant significance (p <0.001, p <0.01).

After 15 days of 1% acetic acid administration, the mean time to enter the dark room was 79.6 seconds in the passive evacuation ability of the control group. In case of beta amyloid group, the beta amyloid group had an average stay of 6.2 seconds (p <0.001). Compared to the dementia group, the mean time to enter the dark room was 91.4 seconds and the mean time to stay 2.6 seconds, which was significantly increased (p <0.001). Among the passive evacuation ability of NP2 administration group, the time to enter the dark room was 81.2 seconds on average, and the retention time was 6.4 seconds on average.

Experimental Example 6: Effects of herbal extracts on animal behavior of dementia animals model (spatial perception test)

Experimental Example 6-1: Spatial Perception Capacity Test (Underwater Maze Test)

Four symbols of O, A, B, and C were attached at equal intervals to the white circular grass having a diameter of 100 cm and a height of 30 cm, respectively, and a platform was under the double O. Water was filled up to 1 cm above the platform (water temperature 15 ° C.) and the water was clouded using prim. At the A, B or C point, the rats were placed in the water and the rats were trained to go to the platform and escape from the water. Training takes place for two weeks and i.c.v. In the group that started training 2 days after injection and the group that started training 10 days after injection, trials for measuring escape latency (triaing trials) were performed. The escape time is the time until the rat climbs onto the platform.

After 24 weeks of training at the end of the two weeks, a probe test was performed to measure memory. Probe testing is performed after the platform has been removed, at which point the first escape latency (FEL) at the point of departure was reached, the second escape latency (SEL) was once again at the platform, and the platform was left for 60 seconds. Platform zone latency (PZL) was measured.

Experimental Example 6-2: Results of underwater-maze test (training trial) of herbal extracts on dementia animal model

For the licorice (NP1) and Earl (NP2) extracts, an underwater-maze experiment (training trial) was conducted to search for the learning and spatial cognitive ability of the dementia animal model. The sample was treated by the method of Experimental Example 3-2, 10 black papers were used, and the results are shown in Tables 5 to 10.

In order to detect the effects of acute dementia and short-term herbal administration, we trained to escape the platform for 14 days with 2 days (2PI) of 1% acetic acid and beta amyloid peptide as icv. . Underwater escape time was 7 seconds for the control group and 14 days for the average 3 seconds when starting from the A point on the right side of the platform. The average of 14 days was 9 seconds, indicating that the acute dementia animal model was produced due to significant delay in underwater escape time on days 1, 3, 4, 5, 6, 7, 8, 9, 10 and 14, respectively. . In the NP1 group, the mean time was 24 seconds and 14 days were the average of 3 seconds. The time to escape was significantly faster from 3 to 14 days than in the dementia group, and the NP2 group was also significantly increased from 2 to 14 days. It was found that the time to escape underwater was faster (Table 5).

Aβ _1-42 Influence of NP1 and NP2 on Escape Time in the Underwater Maze Test (Training Trial) of Rats Treated with (Departure from A, 2PI) Days of training  Escape Time (sec) Mean ± S.E.M. Control Ab NP1 NP2 One 7 ± 2.17 28 ± 20.11 ^## 24 ± 18.01 6 ± 2.14 2 8 ± 4.82 10 ± 5.91 10 ± 4.92 4 ± 0.81 ^** 3 3 ± 0.43 9 ± 3.8 ^### 3 ± 0.54 ^*** 3 ± 0.54 ^*** 4 3 ± 0.43 5 ± 1.11 ^### 4 ± 1.28 ^** 3 ± 0.83 ^*** 5 4 ± 2.35 11 ± 8.78 ^# 3 ± 0.94 ^* 3 ± 0.94 ^* 6 8 ± 3.03 10 ± 6.69 ^# 6 ± 4.06 ^*** 7 ± 3.58 ^*** 7 3 ± 0.01 7 ± 7.43 ^# 4 ± 1.11 ^* 4 ± 1.00 ^* 8 3 ± 1.00 9 ± 4.89 ^## 3 ± 0.63 ^** 3 ± 0.64 ^** 9 8 ± 2.68 5 ± 1.1 ^# 4 ± 0.5 ^** 3 ± 0.30 ^*** 10 3 ± 0.01 4 ± 0.94 ^## 3 ± 0.92 ^** 3 ± 0.67 ^*** 11 4 ± 0.87 5 ± 1.75 3 ± 0.46 ^* 3 ± 1.37 ^** 12 4 ± 1.73 6 ± 1.60 4 ± 0.94 ^** 3 ± 1.02 ^*** 13 4 ± 1.92 5 ± 2.71 3 ± 0.49 ^* 3 ± 1.10 ^* 14 3 ± 0.43 9 ± 6.56 ^## 3 ± 0.70 ^* 3 ± 0.45 ^*

###: P <0.001, ##: P <0.01, #: P <0.05, compared with control

***: P <0.001, **: P <0.01, *: P <0.05, compared with Aβ1-42 treatment group

 When starting from point B on the opposite side of the platform, the underwater escape time was significantly delayed in the beta amyloid group except 12 and 13 days, indicating that it was a dementia animal, and both NP1 and NP2 groups were dementia animals. Underwater escape times were significantly faster in all days 1 to 14 (Table 6).

Aβ _1-42 Influence of NP1 and NP2 on Escape Time in the Underwater Maze Test (Training Trial) of Rats Treated (Departure from B, 2PI) Days of training Escape Time (sec) Average ± S.E.M.  Control Ab NP1 NP2 One 13 ± 6.91 64 ± 4.5 ^### 40 ± 22.16 ^** 5 ± 1.97 ^*** 2 7 ± 2.68 33 ± 16.78 ^### 7 ± 4.05 ^*** 7 ± 3.7 ^*** 3 4 ± 0.71 8 ± 4.36 ^## 7 ± 5.35 4 ± 1.18 ^** 4 6 ± 0.83 9 ± 3.16 ^## 4 ± 1.56 ^*** 8 ± 3.23 5 9 ± 6.22 11 ± 7.68 5 ± 2.71 ^* 5 ± 2.5 ^* 6 4 ± 1.09 22 ± 11.36 ^### 9 ± 6.1 ^** 17 ± 9.62 7 10 ± 6.87 8 ± 4.41 ^# 4 ± 0.87 ^** 4 ± 2.01 ^* 8 4 ± 1.3 12 ± 10.26 ^## 5 ± 1.73 ^* 5 ± 2.2 ^* 9 4 ± 0.87 8 ± 4.92 ^# 4 ± 1.02 ^* 3 ± 0.87 ^** 10 4 ± 1.5 4 ± 1.74 ^# 3 ± 0.66 ^*** 3 ± 0.8 ^*** 11 5 ± 1.92 12 ± 10.91 ^# 6 ± 2.01 ^* 4 ± 1.47 ^* 12 3 ± 0.00 10 ± 8.33 3 ± 0.92 ^* 4 ± 0.92 ^* 13 8 ± 2.69 5 ± 1.72 4 ± 1.28 ^* 4 ± 1.62 14 3 ± 1 6 ± 4.15 ^# 4 ± 1.2 ^* 3 ± 1.04 ^*

###: P <0.001, ##: P <0.01, #: P <0.05, compared with control

***: P <0.001, **: P <0.01, *: P <0.05, compared with Aβ1-42 treatment group

Starting from point C on the left side of the platform, both the NP1 and NP2 groups were significantly faster than the demented animals in the time of escape (Table 7).

Aβ _1-42 Influence of NP1 and NP2 on Escape Time in Underwater Maze Tests (Training Trials) of Rats Treated (Departing from C, 2PI) Days of training Escape Time (sec) Mean ± S.E.M. Control Ab NP1 NP2 One 6 ± 2.55 38 ± 15.44 ^### 12 ± 7.28 ^** 11 ± 5.68 ^*** 2 5 ± 1.50 24 ± 11.25 ^### 4 ± 1.18 ^*** 5 ± 1.69 ^*** 3 4 ± 1.64 9 ± 5.55 ^# 6 ± 3.36 3 ± 0.78 ^** 4 7 ± 4.49 8 ± 3.71 4 ± 0.66 ^*** 4 ± 1.58 5 9 ± 8.26 17 ± 11.25 3 ± 1.00 ^* 4 ± 1.60 ^* 6 7 ± 1.48 15 ± 16.46 ^## 5 ± 1.78 ^** 6 ± 4.36 7 5 ± 2.60 8 ± 5.86 ^# 4 ± 0.95 ^** 5 ± 2.41 ^* 8 4 ± 2.17 11 ± 8.4 ^# 4 ± 1.62 ^* 5 ± 1.28 ^* 9 4 ± 0.87 12 ± 12.29 ^# 4 ± 0.83 ^* 4 ± 1.00 ^** 10 4 ± 0.87 7 ± 4.21 ^# 3 ± 0.45 ^*** 4 ± 0.81 ^*** 11 4 ± 2.17 9 ± 6.39 4 ± 1.22 ^* 4 ± 1.00 ^* 12 3 ± 0.43 9 ± 7.19 ^# 4 ± 1.00 ^* 4 ± 1.36 ^* 13 5 ± 0.87 5 ± 1.27 3 ± 0.54 ^* 4 ± 0.80 14 4 ± 0.50 7 ± 1.28 ^### 4 ± 0.83 ^* 3 ± 0.75 ^*

###: P <0.001, ##: P <0.01, #: P <0.05, compared with control

***: P <0.001, **: P <0.01, *: compared with P <0.05 Aβ1-42 treatment group

To investigate the efficacy of chronic dementia and herbal long-term administration, animals were subjected to maze experiments in water for 14 days with 10 days (10PI) of 1% acetic acid and beta amyloid peptide administered as i.c.v. Underwater escape time was 7 seconds in the control group and 14 days in the average 3 seconds when starting from the A point on the right side of the platform. The escape time was gradually decreased in the beta amyloid group. The mean 14-day averaged 7 seconds, which significantly delayed the escape time at 1, 2, 3, 4, 7, 8, 10, 11, 12, 13, and 14 days, respectively, compared to the control group. Could. In the NP1-treated group, mean 1-day 42-second and 14-day 4-average mean significantly longer underwater escape times at 1, 4, 6, 7, 8, 9, 11, 12, 13, and 14 days compared to chronic dementia animals. This is faster. The NP2 administration group also showed a significantly faster underwater escape time at 2, 7, 8, 9, 11, 12, and 13 days (Table 8).

Aβ _1-42 Influence of NP1 and NP2 on Escape Time in the Underwater Maze Test (Training Trial) of Rats Treated (Departure from A, 10PI) Days of training Escape Time (sec) Mean ± S.E.M. Control Ab NP1 NP2 One 7 ± 2.17 58 ± 8.40 ^### 42 ± 16.84 ^* 58 ± 10.50 2 8 ± 4.82 21 ± 17.16 ^# 10 ± 8.31 10 ± 5.58 ^* 3 3 ± 0.43 8 ± 2.54 ^### 6 ± 3.29 6 ± 3.03 4 3 ± 0.43 17 ± 16.66 ^# 6 ± 2.61 ^* 9 ± 4.38 5 4 ± 2.35 14 ± 17.21 4 ± 1.48 4 ± 0.78 6 8 ± 3.03 10 ± 10.90 3 ± 0.60 ^* 8 ± 4.15 7 3 ± 0.00 16 ± 11.60 ^## 5 ± 2.21 ^** 4 ± 0.78 ^** 8 3 ± 1.00 12 ± 3.16 ^### 5 ± 1.78 ^*** 6 ± 2.80 ^*** 9 8 ± 2.68 8 ± 3.17 4 ± 1.74 ^** 3 ± 0.78 ^*** 10 3 ± 0.00 12 ± 3.67 ^### 7 ± 7.70 12 ± 8.07 11 4 ± 0.87 14 ± 9.32 ^## 4 ± 1.20 ^** 4 ± 1.38 ^** 12 4 ± 1.73 23 ± 11.23 ^### 3 ± 0.45 ^*** 5 ± 1.90 ^*** 13 4 ± 1.92 8 ± 3.64 ^# 3 ± 0.60 ^** 3 ± 0.63 ^*** 14 3 ± 0.43 7 ± 2.40 ^### 4 ± 0.98 ^** 6 ± 2.00

###: P <0.001, ##: P <0.01, #: P <0.05 Comparison with Control

***: P <0.001, **: P <0.01, *: compared with P <0.05 Aβ1-42 treatment group

 Starting from point B, on the other side of the platform, the underwater escape time was significantly delayed except for 4, 5, 7, and 13 days for the beta amyloid group, indicating that it was a chronic dementia animal. In both NP1 and NP2 groups, the escape time was significantly faster from 1 to 14 days compared to the chronic dementia group (Table 9).

Aβ _1-42 Effect of NP1 and NP2 on Escape Time in Underwater Maze Tests (Training Trials) of Rats Treated (Departure from B, 10PI) Training Days Escape Time (sec) Mean ± S.E.M. Control Ab NP1 NP2 One 13 ± 6.91 61 ± 7.17 ^### 49 ± 19.21 42 ± 12.94 ^** 2 7 ± 2.68 37 ± 16.06 ^### 23 ± 20.91 45 ± 20.79 3 4 ± 0.71 34 ± 21.79 ^## 7 ± 2.84 ^** 11 ± 8.39 ^** 4 6 ± 0.83 15 ± 17.09 10 ± 6.45 13 ± 4.64 5 9 ± 6.22 18 ± 16.60 5 ± 5.16 ^* 14 ± 6.45 6 4 ± 1.09 14 ± 9.95 ^## 4 ± 1.78 ^* 6 ± 2.87 ^* 7 10 ± 6.87 8 ± 4.88 5 ± 1.75 ^* 5 ± 1.28 ^* 8 4 ± 1.3 18 ± 13.29 ^## 5 ± 2.2 ^* 9 ± 5.31 ^* 9 4 ± 0.87 5 ± 2.04 ^# 5 ± 2.02 ^* 4 ± 1.25 ^* 10 4 ± 1.5 25 ± 8.33 ^### 5 ± 1.84 ^** 6 ± 1.86 ^*** 11 5 ± 1.92 22 ± 12.04 ^### 5 ± 2.69 ^** 5 ± 3.81 ^*** 12 3 ± 0.00 15 ± 17.19 ^# 4 ± 1.57 ^*** 4 ± 1.02 ^* 13 8 ± 2.69 10 ± 3.77 5 ± 2.50 ^** 5 ± 1.73 ^** 14 3 ± 1.00 13 ± 6.74 ^### 7 ± 5.64 ^* 6 ± 1.96 ^**

###: P <0.001, ##: P <0.01, #: P <0.05 Comparison with Control

***: P <0.001, **: P <0.01, *: compared with P <0.05 Aβ1-42 treatment group

Starting from point C on the left side of the platform, both the NP1 and NP2 administration groups showed significantly faster underwater escape time than the demented animals (Table 10).

Aβ _1-42 Effect of NP1 and NP2 on Escape Time in Underwater Maze Tests (Training Trials) of Rats Treated (Departing from C, 10PI) Days of training  Escape Time (sec) Mean ± S.E.M. Control Ab NP1 NP2 One 6 ± 2.55 61 ± 7.19 ^### 52 ± 14.61 58 ± 10.50 2 5 ± 1.50 31 ± 21.65 ^## 12 ± 7.97 ^* 21 ± 15.98 3 4 ± 1.64 21 ± 18.91 ^# 6 ± 2.48 ^* 9 ± 3.17 ^* 4 7 ± 4.49 22 ± 9.59 ^## 7 ± 2.32 ^*** 11 ± 5.87 ^** 5 9 ± 8.26 24 ± 13.64 ^# 6 ± 3.12 ^** 6 ± 3.42 ^** 6 7 ± 1.48 18 ± 17.06 ^# 5 ± 1.55 ^* 5 ± 1.78 ^* 7 5 ± 2.60 11 ± 4.37 ^## 7 ± 2.46 ^* 6 ± 1.81 ^** 8 4 ± 2.17 17 ± 15.98 ^# 7 ± 2.51 ^* 5 ± 2.59 ^* 9 4 ± 0.87 8 ± 4.26 ^## 4 ± 1.19 ^* 5 ± 1.61 ^* 10 4 ± 0.87 24 ± 12.38 ^### 7 ± 4.36 ^** 8 ± 2.20 ^** 11 4 ± 2.17 23 ± 17.71 ^## 3 ± 0.70 ^** 4 ± 1.11 ^** 12 3 ± 0.43 20 ± 9.76 ^### 3 ± 0.40 ^*** 4 ± 1.18 ^*** 13 5 ± 0.87 7 ± 2.20 ^# 4 ± 0.63 ^** 4 ± 1.56 ^* 14 3 ± 0.50 8 ± 3.73 ^## 5 ± 1.41 ^* 4 ± 0.98 ^**

###: P <0.001, ##: P <0.01, #: P <0.05 Comparison with Control

***: P <0.001, **: P <0.01, *: compared with P <0.05 Aβ1-42 treatment group

Experimental Example 6-3: underwater-maze test (probe test) results of herbal extracts for dementia animal model

In order to test the learning and spatial perception of the animals trained through the underwater maze training trial experiment for 14 days, the platform was removed and the underwater maze probe test was performed.

In case of starting from point A, the 2PI animal group had an average 19 second escape time in the control group, the 2nd average 26 seconds, and the PZ averaged 7 seconds, whereas the beta amyloid group had an average of 40 seconds for the primary and 50 seconds for the secondary. , PZ was found to be 4 seconds, indicating that acute dementia animals were produced.In the NP1 and NP2 administration groups, EL-1 averaged 11, 9 seconds, and EL-2 averaged 21, 24, respectively. Second, PZ was 9 seconds and 7 seconds on average, respectively. In the case of 10PI, a chronic dementia animal model, the NP1 and NP2 administration groups showed significant (p <0.001) therapeutic effects (Table 11).

Aβ _1-42 Influence of NP1 and NP2 on the escape time and platform point residence time (departure from A) in the underwater maze test (probe test) of rats treated with group Time (seconds) Mean ± S.E.M. EL-1 EL-2 PZ 2PI Control 19 ± 12.93 26 ± 15.28 7 ± 3.5 Ab 40 ± 16.51 ^# 50 ± 9.94 ^# 4 ± 1.55 NP1 11 ± 7.93 ^*** 21 ± 9.64 ^*** 9 ± 2.79 ^*** NP2 9 ± 4.87 ^*** 24 ± 13.55 ^*** 7 ± 2.33 ^** 10PI Control 19 ± 12.93 26 ± 15.28 7 ± 3.5 Ab 52 ± 15.62 ^## 59 ± 13.29 ^# 4 ± 1.38 NP1 10 ± 5.34 ^*** 20 ± 8.57 ^*** 10 ± 3.20 ^*** NP2 9 ± 6.00 ^*** 14 ± 4.82 ^*** 9 ± 2.34 ^***

##: P <0.01, #: P <0.05 compared to control

***: P <0.001, **: compared with P <0.01 Aβ1-42 treatment group

In the case of starting at point B, the administration of NP1 and NP2 also improved remarkably (p <0.001-p <0.01), which improved learning and spatial perception ability (Table 12). Significant (p <0.001-p <0.01) learning and spatial perception skills were found to improve (Table 13).

Aβ _1-42 Influence of NP1 and NP2 on escape time and platform point residence time (departure from B) in an underwater maze test (probe test) of rats treated with group  Time (sec) Average ± S.E.M. EL-1 EL-2 PZ 2PI Control 5 ± 2.38 10 ± 2.35 12 ± 1.48 Ab 14 ± 11.34 ^### 37 ± 14.19 ^# 6 ± 2.96 ^### NP1 10 ± 8.1 ^* 24 ± 10.94 7 ± 3.41 NP2 12 ± 1059 ^** 19 ± 8.29 6 ± 1.87 10PI Control 5 ± 2.28 10 ± 2.35 12 ± 1.48 Ab 39 ± 20.63 ^### 50 ± 17.64 ^### 5 ± 2.25 ^### NP1 6 ± 2.32 ^*** 14 ± 4.84 ^*** 10 ± 3.25 ^*** NP2 6 ± 1.75 ^*** 12 ± 4.21 ^*** 8 ± 2.23 ^**

##: P <0.01, #: P <0.05 compared to control

***: P <0.001, **: compared with P <0.01 Aβ1-42 treatment group

Aβ _1-42 Effect of NP1 and NP2 on Escape Time and Platform Point Retention Time in the Underwater Maze Test (Probe Test) of Rats Treated (Departing from C) group Time (sec) Mean ± S.E.M. EL-1 EL-2 PZ 2PI Control 5 ± 0.5 14 ± 3.35 9 ± 1.48 Ab 30 ± 19.03 ^### 49 ± 13.39 ^## 4 ± 2.14 ^## NP1 21 ± 17.84 35 ± 19.12 5 ± 3.45 NP2 14 ± 11.03 ^*** 26 ± 12.95 ^* 5 ± 1.7 10PI Control 5 ± 0.5 14 ± 3.35 9 ± 1.48 Ab 43 ± 15.94 ^### 52 ± 12.04 ^### 6 ± 1.97 ^# NP1 13 ± 7.65 ^*** 24 ± 15.03 ^*** 9 ± 3.71 ^* NP2 7 ± 2.00 ^*** 12 ± 3.72 ^*** 8 ± 2.49 ^*

##: P <0.01, #: P <0.05 compared to control

***: P <0.001, **: compared with P <0.01 Aβ1-42 treatment group

All results of the experimental example was expressed as the average ± S.E.M value and the statistical significance was verified through Student's t-test.

The present invention provides a pharmaceutical composition or functional food composition for the prevention or treatment of dementia, including licorice, earl pear, Angelica, bark, sanjoin, lettuce, changchangpo, horse riding, oak, and gardenia extract. In addition, the present invention provides a pharmaceutical composition or functional food composition for improving cognitive function, including licorice, earl pear, Angelica, bark skin, Sanjoin, Sangbaekpi, Seokchangpo, horse riding, oak and gardenia extract. The extract has an inhibitory activity of inhibiting acetylcholinesterase or beta amyloid aggregation, thereby exhibiting an excellent effect of preventing or treating dementia and improving cognitive function. In particular, the licorice, earl, lychee, horse riding and ointment extracts of the herbal extracts have excellent neuronal protection and are more useful for preventing or treating dementia and improving cognitive function. According to the animal efficacy test for licorice and earl herb, the two herbal extracts show excellent efficacy of improving the learning and spatial perception ability of the dementia animal group induced by beta amyloid close to the normal animal group. In addition, since the composition of the present invention contains the herbal extract as an active ingredient, side effects on the human body are extremely less than that of the chemical synthetic medicine.

Claims (4)

(a) a pharmaceutically effective amount of one or more herbal extracts of licorice, sanjoin and oak extracts; And (b) a pharmaceutically acceptable carrier. delete (a) a pharmaceutically effective amount of one or more herbal extracts of licorice, sanjoin and oak extracts; And (b) a pharmaceutical composition for improving cognitive function comprising a pharmaceutically acceptable carrier. delete