KR20120025826A - Composition comprising ligularia fischeri extract for protecting nerve cells - Google Patents

Abstract

The present invention provides a brain nerve cell protective use of the bear extract or active ingredient 2,5,8,11,14,17,20-heptaoxahenekosan isolated therefrom. These components exhibit excellent oxidative stress inhibitory activity and protective effect on brain neurons. Therefore, the composition comprising them as an active ingredient, cerebral nerve disease, Alzheimer's disease, Parkinson's disease, cerebral amyloid angiopathy ( Cerebral amyloid angiopathy ), dementia and Huntington's disease can be used to effectively prevent, treat or ameliorate various diseases.

Description

Composition comprising Ligularia fischeri extract for protecting nerve cells

The present invention relates to a brain nerve cell protective composition comprising a bear extract or 2,5,8,11,14,17,20-heptaoxaheneko acid as an active ingredient.

Amyloid beta (amyloid β, Aβ) is a 40-42 peptide produced by abnormal cleavage of amyloid precursor protein (APP) and is an important pathogen in brain neurological diseases including Alzheimer's disease (AD). It is considered. The cause of neural cell necrosis in AD is still unclear, but the generation and production of oxygen free radicals and oxidative injury such as reactive oxygen species (ROS) can lead to the destruction and development of cerebral nerve cells. It is presumed to be related to pathogenesis. Increased ROS (lipid peroxidation, protein modification, mitochondrial DNA oxidation) have been demonstrated in the brains of AD patients, and these physiological metabolic abnormalities include amyloid plaques and neurofibrillary tangles (NFTs). The same AD brain results in unique clinical results. In addition, since Aβ induces intra-cellular free radical status and ultimately induces cell necrosis, Aβ and brain nerve cell destruction appear to be correlated. The production of ROS, free radicals, etc. by A [beta] enables the estimation of A [beta] -induced free radical-mediated neurotoxicity. .

To date, the main targets for AD drug development are cholinesterase inhibitors (Aricept, Exelon, Reminyl, etc.) and glutamate antagonists that target cholinergic neurons beyond disease neurotransmitters. However, since these drugs only temporarily relieve symptoms, there is an urgent need for drug development techniques that treat the underlying disease or inhibit the progression of the disease itself.

The present invention to develop a substance that can inhibit the toxicity of the brain neurons due to oxidative stress induced by amyloid beta from edible biological resources, that is, to show the brain neuron cell protection, to provide a composition for protecting the brain neurons comprising the same For the purpose of

In addition, an object of the present invention is to provide a method for purifying the brain neuronal cell protective material from edible biological resources.

To this end, the inventors of the present inventors have studied whether the use of food for a long time to inhibit the toxicity of cerebral neurons caused by amyloid beta-induced oxidative stress in edible biological resources, it was confirmed that the bear extract has this effect .

Therefore, the present invention provides a composition for protecting brain neurons comprising the bear extract as an active ingredient.

In the preparation of the bear odor extract of the present invention, dried or undried bear odor or a mixture thereof can be used. For efficiencies of extraction, the odor can be finely used. In addition, in order to obtain an effective brain nerve cell protective bear odor extract, it may be subjected to a process of incubating for about 5 minutes at a high temperature of about 100 ℃ before grinding the bear odor to inactivate the intracellular enzymes.

For the production of bear odor extract can be extracted according to a conventional extraction method using 3 to 10 times the extraction solvent of the bear odor.

The extraction solvent may be extracted using water, an organic solvent or a mixed solvent thereof which is widely used in natural product extraction. The organic solvent includes ethanol, methanol, hexane, chloroform, etaacetate and the like. Such organic solvents may be used in admixture with water, but may be used without mixing with water for easy elution of the active ingredient. In one embodiment, the bear extract may be an ethanol extract.

The bear extract can be further fractionated using an organic solvent that is different in polarity from the organic solvent used in the first extraction. In this case, the bear odor extract may be a bear odor extract fraction obtained by extracting the bear odor with an organic solvent and fractionating the residue using an organic solvent having a different polarity.

In addition, the Goji odor extract or Goji odor extract fractions are purified using various chromatography such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography, and the like. It is also possible to obtain further purified fractions. In the present invention, the 'bear's extract' includes all extracts, fractions and purified products obtained in each step of extraction, fractionation or purification as described above, their dilutions, concentrates or dried products.

In one embodiment of the present invention, the bear extract may be a chloroform fraction obtained by extracting the bear odor with ethanol, the residue is sequentially fractionated with hexane, chloroform and ethyl acetate.

In another embodiment of the present invention, the bear extract is an extract of the bear odor with ethanol, and the chloroform fraction obtained by sequentially fractionating the residue with hexane, chloroform and ethyl acetate using a mixed solvent of chloroform and ethanol as a developing solvent. In the fraction obtained by performing silica gel column chromatography, chloroform: ethanol = 98: 2 ~ 90:10 may be the extract of Goji odor extract obtained under the developing solvent conditions.

In another embodiment of the present invention, the bear extract is extracted from bear ethanol with ethanol, and the chloroform fraction obtained by sequentially fractionating the residue with hexane, chloroform and ethyl acetate using a mixed solvent of chloroform and ethanol as a developing solvent. In the fraction obtained by performing silica gel column chromatography, chloroform: ethanol = 94: 6 may be a fraction of the extract from the odor obtained under the developing solvent conditions.

The purified polar bear extract fraction may be further subjected to repeated column chromatography or thin layer chromatography (TLC) or high performance liquid chromatography (HPLC).

In the following example, the active ingredient showing the neuroprotective activity of the brain cells from the odor extract through this additional purification process 2,5,8,11,14,17,20-heptaoxahenenoic acid (2,5,8,11, 14,17,20-heptaoxaheneicosane) was confirmed.

Accordingly, the present invention provides a composition for protecting nerve cells comprising 2,5,8,11,14,17,20-heptaoxahenecaic acid extract as an active ingredient.

The invention also

1) extracting the bear odor with an organic solvent, water or a mixed solvent thereof, and fractions of the residue were sequentially extracted using hexane, chloroform and ethyl acetate to obtain a chloroform fraction;

2) The chloroform fraction obtained in step 1 was subjected to silica gel column chromatography using a mixed solvent of chloroform and ethanol as a developing solvent. Chloroform in the fraction obtained from methanol: 98: 2 ~ 90:10. Obtaining an extract fraction

Provided is a method for purifying the bear odor extract fraction.

The present invention also provides a composition for protecting nerve cells comprising 2,5,8,11,14,17,20-heptaoxaheneko acid as an active ingredient. Although 2,5,8,11,14,17,20-heptaoxahenekoic acid is isolated from the bear odor extract according to the present invention, it can be chemically synthesized or commercially available and is also isolated from other plants. You can do it.

The invention also

1) extracting the bear odor with an organic solvent, water or a mixed solvent thereof, and fractions of the residue were sequentially extracted using hexane, chloroform and ethyl acetate to obtain a chloroform fraction;

2) The chloroform fraction obtained in step 1 was subjected to silica gel column chromatography using a mixed solvent of chloroform and ethanol as a developing solvent. Chloroform in the fraction obtained from methanol: 98: 2 ~ 90:10. Obtaining an extract fraction and

3) purifying the bear extract extract showing the brain nerve cell protective activity by performing HPLC (high performance liquid chromatography) on the bear extract extract obtained from step 2

Containing

Provided are methods for purifying 2,5,8,11,14,17,20-heptaoxahenechoic acid (2,5,8,11,14,17,20-heptaoxaheneicosane).

In the following example of the present invention, the residue obtained by extracting Gom odor with 100% ethanol, fractionated in the order of hexane, chloroform, ethyl acetate in order to prepare a Gom odor extract fraction. In addition, the chloroform fraction of the bear extract fractions were further fractionated and purified. Bear extract, bear extract fraction, and 2,5,8,11,14,17,20-heptaoxahenenoic acid of the present invention obtained through this process shows an excellent oxidative stress inhibitory effect and brain neuronal cell protective effect.

Therefore, bear odor extract, bear odor extract fraction, and 2,5,8,11,14,17,20-heptaoxahenenoic acid of the present invention may be included as an active ingredient of the composition for protecting brain neurons.

The composition for protecting brain neurons of the present invention can be used as a pharmaceutical composition for the prevention and treatment of diseases caused by the toxicity of brain neurons due to oxidative stress induced by amyloid beta. In addition, the composition for protecting brain neurons of the present invention can be used as a food composition for improving diseases caused by the toxicity of brain neurons due to oxidative stress induced by amyloid beta.

Diseases caused by the toxicity of cerebral neurons due to oxidative stress induced by amyloid beta are not limited to, for example, Alzheimer's disease (AD), Parkinson's disease (PD), dementia (dementia), amyotrophic lateral sclerosis (ALS), corticobasal degeneration (CBD), multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Huntington's disease (HD), cerebral amyloid angiopathy ( Cerebral) amyloid angiopathy ), and the like.

The composition can be used with not only the active ingredient of the present invention but also known neuroprotective neurons.

The pharmaceutical composition comprising bear extract of the present invention or 2,5,8,11,14,17,20-heptaoxaheneko acid as an active ingredient is a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient. It may be prepared using, and the auxiliary agent may be used excipients, disintegrants, sweeteners, binders, coatings, expanding agents, lubricants, lubricants or flavoring agents.

The pharmaceutical composition may be preferably formulated into a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers in addition to the active ingredient described above for administration.

Formulation forms of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include but are not limited to natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, corn sweeteners, acacia, trackercance or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can be formulated according to each disease or component, as appropriate in the art.

The present invention also provides a food composition for brain nerve cell protection comprising the bear extract or 2,5,8,11,14,17,20-heptaoxaheneko acid.

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

The present invention provides a health functional food comprising the composition for protecting the brain neurons.

The present invention also provides the use of the above-mentioned bear extract or a composition comprising 2,5,8,11,14,17,20-heptaoxahenenoic acid as an active ingredient for the manufacture of a medicament or food for protecting brain nerve cells. The composition of the present invention comprising the above-mentioned bear extract or 2,5,8,11,14,17,20-heptaoxaheneko acid as an active ingredient is a pharmaceutical or food for the prevention or treatment of diseases related to neuroprotective neurons It can be used for manufacturing purposes.

The present invention also relates to a method for protecting nerve cells comprising administering to a mammal a therapeutically effective amount of a bear extract or 2,5,8,11,14,17,20-heptaoxahenecaic acid, or a disease associated with brain nerve cell protection. It provides a method of preventing or treating.

As used herein, the term "mammal" refers to a mammal that is the subject of treatment, observation or experimentation, preferably human.

As used herein, the term “therapeutically effective amount” means an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, doctor or other clinician, This includes amounts that induce alleviation of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dosages and frequency of administrations for the active ingredients of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be readily determined by one skilled in the art and includes the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, general health of the patient. It may be adjusted according to various factors including the condition, sex and diet, time of administration, route of administration and the rate of secretion of the composition, the duration of treatment, and the drugs used simultaneously. In the treatment method of the present invention, in the case of an adult, 1 mg / time of once or several times daily administration of bear extract of the present invention or 2,5,8,11,14,17,20-heptaoxaheneko acid It is preferable to administer at a dose of kg-1000 mg / kg.

In the treatment method of the present invention, the present invention comprises a bear odor extract or a composition comprising 2,5,8,11,14,17,20-heptaoxahenenoic acid as an active ingredient, oral, rectal, intravenous, intraarterial, abdominal cavity. Administration can be in conventional manner via the intra, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal route.

The present invention provides a brain nerve cell protective use of the bear extract or active ingredient 2,5,8,11,14,17,20-heptaoxahenekosan isolated therefrom. These components exhibit excellent oxidative stress inhibitory activity and protective effect on brain neurons. Thus, compositions comprising them as active ingredients include Alzheimer's disease (AD), Parkinson's disease (PD), dementia, amyotrophic lateral sclerosis (ALS), and cortical hypodegeneration (ALS). corticobasal degeneration (CBD), multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Huntington's disease (HD), and cerebral amyloid angiopathy ( Cerebral) amyloid It can be used to effectively prevent, treat or ameliorate various diseases such as angiopathy ).

Figure 1 is a graph showing the cell survival rate of bear ethanol extract treatment group.
Figure 2 is a graph showing the oxidative stress inhibitory activity of the organic solvent fraction treatment group of Bear ethanol extract.
Figure 3 is a graph showing the cell viability of the organic solvent fraction treatment group of Bear ethanol extract.
FIG. 4 is a graph showing the oxidative stress inhibitory activity of the first open silica gel column chromatography fraction treatment group of goji chloroform fraction (chlo3).
Figure 5 is a graph showing the cell viability of the first open silica gel column chromatography fraction treatment group of goji chloroform fraction (chlo3).
Figure 6 is a small fraction obtained by the first open silica gel column chromatography (CHCl ₃ : EtOH = 90:10 (v / v) Graph showing the oxidative stress inhibitory activity of the fraction treated group obtained by performing the second open silica gel column chromatography on the first fraction (fraction 4)).
Figure 7 is a small fraction obtained by the first open silica gel column chromatography (CHCl ₃ : EtOH = 90:10 (v / v) Graph showing the cell viability of the fraction treatment group obtained by performing the second open silica gel column chromatography on the first fraction (fraction 4)).
8 is a graph showing the HPLC results of the active fraction (fraction 4; chloroform: ethanol = 94: 6 (v / v)) separated from the second open silica gel column chromatography.
9 is a table summarizing the purification process of the bear odor extract through Examples 1 to 3.
10 is a schematic diagram of an in vivo experiment for evaluating the neuronal neuroprotective activity of bear extract.
FIG. 11 is a result of the Y-maze test showing that the memory learning ability decreased by injection of Aβ was increased again in the group receiving the odor extract.
12 is the result of a passive avoidance test showing that the memory learning ability reduced by injection of Αβ increased again in the group receiving the odor extract.

Advantages and features of the present invention and methods for achieving them will be apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in various forms, and only the embodiments are intended to complete the disclosure of the present invention, and the general knowledge in the technical field to which the present invention pertains. It is provided to fully convey the scope of the invention to those skilled in the art, and the present invention is defined only by the scope of the claims.

 [ Example ]

Example  1: Preparation of Bear Extract and Nerve cells  Check protection activity

After finely grinding the bear odor, 2.3 kg of bear odor was shaken (shaking) for 5 hours with 5 times the volume of ethanol (v / v) of the sample for 5 hours and then concentrated under reduced pressure to prepare a bear ethanol extract.

In order to confirm whether the ethanol extract of bear odor produced in this manner exhibited neuroprotective activity, the neuronal necrosis inhibition activity of the odor ethanol extract was measured.

First, PC12 cells were cultured in RPMI-1640 medium containing antimicotics / antibiotics until the number of cells reached 10 ⁴ -10 ⁶ cells / ml. The control was treated with deionized water instead of H ₂ O ₂ . The negative control was treated with 200 μM of H ₂ O ₂ except for inducing oxidative stress. Positive controls were pre-cultured in RPMI-1640 medium containing 100 μM Vit C for 48 hours before treatment with ₂₀₀ μM H ₂ O ₂ . The experimental group was pre-incubated in RPMI-1640 medium containing 1 mg / ml of Sengmul extract for 48 hours before treatment with ₂₀₀ μM of H ₂ O ₂ .

Oxidative injury was measured by spectrofluorometer using DCF-DA (2 ', 7'-dichlorofluorescin diacetate) assay. As a result, as shown in Figure 1, the experimental group treated with the bear ethanol extract showed a protective effect of 81.8% for the brain neurons compared with the group (negative) not treated with the bear ethanol extract. This is the result of checking the cell viability through the MTT reduction assay. The data in FIG. 1 is shown as mean ± SD for four independent experiments.

Example  2: preparation of bear odor extract fraction and Nerve cells  Check protection activity

After completely removing the ethanol from the odor extract ethanol extract prepared in Example 1 and extracted with reflux in three volumes using hexane, chloroform and ethyl acetate each fraction (Hex1, Hex2, Hex3, Chlo1, Chlo2, Chlo3, Ethyl1, Ethyl2, Ethyl3) were prepared.

The hexane fraction, the chloroform fraction and the ethyl acetate fraction of the ethanol extracts of the present invention were measured for oxidative stress inhibition and neuronal necrosis inhibition of these fractions ( 1 mg / ml ).

Neuronal necrosis inhibition was measured in the same manner as in Example 1, and oxidative stress inhibition was measured in the following manner. PC12 cells were cultured in RPMI- 1640 medium containing antimicotics / antibiotics. When the number of cells reached 10 ⁴ -10 ⁶ cells / ml, each of the extracts to be measured for 48 hours was treated with H ₂ O ₂ for oxidation. Induces enemy stress. After 2 hours DCF - was measured by the meter to the oxidative stress level generated by a DA (2 ', 7'-dichlorofluorescin diacetate) spectrometer with fluoro (spectrofluorometer).

As a result, as can be seen in Figures 2 and 3, the third chloroform layer (Chol3) and the first ethyl acetate layer (Ethyl1) showed excellent oxidative stress inhibitory activity and neuronal necrosis inhibitory activity.

33 small fractions (CHCl ₃₎ were subjected to primary open silica gel column chromatography on the Chol3 fraction with the highest inhibitory effect. : EtOH = 100: 0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90, 0: 100 Fractions). For the small fraction of the first open silica gel column chromatography, oxidative stress inhibition and neuronal necrosis inhibition were confirmed by the same method as above.

As a result, CHCl ₃ as can be seen in Figures 4 and 5 : The first subfraction (fraction 4) obtained under the condition of EtOH = 90:10 (v / v) showed the highest inhibitory activity.

CHCl ₃ : EtOH = 90:10 (v / v) Chloroform and ethanol (100: 0, 98: 2, 96: 4, 94: 6) using the second open silica gel column chromatography on the first fraction (vction 4) , 92: 8, 90:10, 88:12, 86:14, 84:16, 82:18, 80:20 v / v), and each of the subfractions obtained from them is oxidatively Stress inhibitory activity and neuronal necrosis inhibition activity were confirmed. As a result, as can be seen in Figures 6 and 7, CHCl ₃ : The fourth small fraction (fraction 4) obtained under the condition EtOH = 94: 6 (v / v) showed the highest inhibitory activity.

Example  3: Confirmation of active substance of bear odor extract fraction

The active fraction (fraction 4; chloroform: ethanol = 94: 6 v / v) isolated from the second open column chromatography was analyzed by HPLC using μ-bondapak ^™ C ₁₈ column. The change was carried out with a linear gradient of 0-100% ethanol and water for 95 minutes. The wavelength range was set to 190-800 nm, the flow rate was 0.5 ml per minute, and 10 μl was injected at a time. The solvent used was ethanol and tertiary distilled water. As a result of the HPLC of FIG. 8, this active fraction has 2,5,8,11,14,17,20-heptaoxahenenoic acid (2,5,8,11,14,17,20) having a molecular weight of 280.14 m / z. -heptaoxaheneicosane) was confirmed.

Summarizing the purification process of the bear odor extract through Examples 1 to 3 is as shown in FIG.

Example  4: of odor extract Nerve cells  For protection active Invivo  Experiment

As shown in FIG. 10, an in vivo experiment was designed to evaluate the neuronal neuroprotective activity of bear extract. First, ICR male mice were fed with adsorbed ethanol extract samples by concentration of ad lib for about 3 weeks (400, 800, 1200 mg / kg, day) by concentration. Intracerebroventricular (ICV) injection of amyloid beta peptide (Aβ), a dementia-causing substance, was performed 21 days after the start of the diet, Y-maze test on day 24 and passive avoidance response test on day 28.

To measure the alteration behavior, conduct a behavioral experiment in a Y-maze with three arms ⓐ, ⓑ and ⓒ respectively. The Y-maze was 32.5 cm long, 15 cm high, and 4 cm wide, with each arm set to A, B, and C, and the arms entered. The mouse is placed in the maze and allowed to move freely for 8 minutes without any irritation. The figure was calculated by measuring two hind feet in the arm except for the tail.

A passive avoidance test was performed to remember the box in which the aversive stimulus (electroshock) was presented and to measure the step-through latency to reenter the same box. The experiment is conducted in the same place as the memory box the day before the actual experiment, and then conducts a behavior experiment. 24 hours after all the mice had undergone adaptive training in the test box on the day before the experiment (Ming-Ming, No-Shock, Shock), with no shock and no light. Experiments were made for 300 sec per head.

As a result of the experiment, as shown in FIGS. 11 and 12, it was confirmed that the reduced memory learning ability (alternation behavior, step-through latency) by injection of Aβ was increased again in the group receiving the bear extract.

Claims (17)

Brain nerve cell protective composition comprising an extract of ligularia fischeri as an active ingredient.
The method of claim 1,
The bear odor extract is a composition for protecting nerve nerve cells of the organic solvent, water or a mixed solvent extract thereof.
The method of claim 1,
The bear odor extract is a ethanol extract brain nerve cell protective composition.
The method of claim 1,
The bear odor extract is a brain odor extract cell extract composition obtained by extracting the bear odor with an organic solvent, fractions obtained by fractionating the residue using an organic solvent having a different polarity.
The method of claim 1,
The bear extract extract is a chloroform fraction obtained by sequentially extract the bear odor with ethanol, the residue of hexane, chloroform and ethyl acetate sequentially fractions.
The method of claim 1,
The bear extract is obtained by extracting the bear odor with ethanol, and performing the silica gel column chromatography using a mixed solvent of chloroform and ethanol as a developing solvent for the chloroform fraction obtained by sequentially fractionating the residue with hexane, chloroform and ethyl acetate. Chloroform in the fraction: Ethanol = 98: 2 ~ 90:10 Brain nerve cell protective composition which is a fraction of the extract obtained from the developing solvent conditions.
The method of claim 1,
The bear extract is obtained by extracting the bear odor with ethanol, and performing the silica gel column chromatography using a mixed solvent of chloroform and ethanol as a developing solvent for the chloroform fraction obtained by sequentially fractionating the residue with hexane, chloroform and ethyl acetate. Chloroform in fractions: Ethanol = brain sperm nerve cell protective composition which is a fraction of the extract of odor obtained under the developing solvent conditions of 94: 6.
The method of claim 1,
The bear odor extract 2,5,8,11,14,17,20-heptaoxaheneko acid (2,5,8,11,14,17,20-heptaoxaheneicosane) composition comprising a protective neurons.
The method according to any one of claims 1 to 8,
The composition is Alzheimer's disease (AD), Parkinson's disease (PD), dementia, amyotrophic lateral sclerosis (ALS), corticobasal degeneration (CBD), Multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Huntington's disease (HD), and cerebral amyloid angiopathy ( Cerebral) amyloid Angiopathy ) A composition for protecting nerve cells, which is a medicament used for the prevention or treatment of diseases selected from the group consisting of.
The method according to any one of claims 1 to 8,
The composition is Alzheimer's disease (AD), Parkinson's disease (PD), dementia, amyotrophic lateral sclerosis (ALS), corticobasal degeneration (CBD), Multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Huntington's disease (HD), and cerebral amyloid angiopathy ( Cerebral) amyloid Angiopathy ) is a food composition used for the improvement of a disease selected from the group consisting of brain nerve cell protective composition.
A health functional food comprising the composition for protecting a brain nerve cell according to any one of claims 1 to 8.
2,5,8,11,14,17,20-heptaoxahenenoic acid (2,5,8,11,14,17,20-heptaoxaheneicosane) A composition for protecting nerve cells comprising as an active ingredient.
The method of claim 12,
The composition is Alzheimer's disease (AD), Parkinson's disease (PD), dementia, amyotrophic lateral sclerosis (ALS), corticobasal degeneration (CBD), Multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Huntington's disease (HD), and cerebral amyloid angiopathy ( Cerebral) amyloid Angiopathy ) A composition for protecting nerve cells, which is a medicament used for the prevention or treatment of diseases selected from the group consisting of.
The method of claim 12,
The composition is Alzheimer's disease (AD), Parkinson's disease (PD), dementia, amyotrophic lateral sclerosis (ALS), corticobasal degeneration (CBD), Multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Huntington's disease (HD), and cerebral amyloid angiopathy ( Cerebral) amyloid Angiopathy ) is a food composition used for the improvement of a disease selected from the group consisting of brain nerve cell protective composition.
A health functional food comprising the composition for protecting a brain nerve cell according to any one of claims 12 to 14.
1) extracting the bear odor with an organic solvent, water or a mixed solvent thereof, and fractions of the residue were sequentially extracted using hexane, chloroform and ethyl acetate to obtain a chloroform fraction; And
2) The chloroform fraction obtained in step 1 was subjected to silica gel column chromatography using a mixed solvent of chloroform and ethanol as a developing solvent. Purifying method of bear extract extract comprising the step of obtaining an extract fraction.
1) extracting the bear odor with an organic solvent, water or a mixed solvent thereof, and fractions of the residue were sequentially extracted using hexane, chloroform and ethyl acetate to obtain a chloroform fraction;
2) The chloroform fraction obtained in step 1 was subjected to silica gel column chromatography using a mixed solvent of chloroform and ethanol as a developing solvent. Chloroform in the fraction obtained from methanol: 98: 2 ~ 90:10. Obtaining an extract fraction and
3) purifying the bear extract extract showing the brain nerve cell protective activity by performing HPLC (high performance liquid chromatography) on the bear extract extract obtained from step 2
Method for purifying 2,5,8,11,14,17,20-heptaoxahenenoic acid (2,5,8,11,14,17,20-heptaoxaheneicosane).

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