KR101770871B1 - Pharmaceutical Composition Comprising Dryopteris crassirhizoma Extract for the Treatment of Degenerative Brain Diseases - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention and treatment of degenerative brain diseases comprising an extract of a spectator as an active ingredient. The pharmaceutical composition for the prevention and treatment of degenerative brain diseases comprising the extract of the present invention as an active ingredient contains the extract extracted from a natural plant as an active ingredient and has an effect of inhibiting the formation of beta-amyloid aggregates. Therefore, Can be used for prevention and treatment of degenerative brain diseases. In addition, since the extract derived from a natural product is an effective ingredient thereof, there is no problem about side effects or cytotoxicity.

Description

TECHNICAL FIELD [0001] The present invention relates to a pharmaceutical composition for preventing and treating degenerative brain diseases,

The present invention relates to a pharmaceutical composition for the prevention and treatment of degenerative brain diseases comprising an extract of a spectator as an active ingredient.

Dementia is a morbid phenomenon distinguishable from normal aging and is distinguished by the cause of Alzheimer's disease, vascular dementia, other alcoholism, trauma, and dementia resulting from Parkinson's disease do.

The mechanism of the onset of Alzheimer's dementia is not clearly known, but the toxicity of the neurotoxic protein, β-amyloid protein, is suggested as the most important cause. This substance is known to be produced by the incorrect metabolism of amyloid precursor protein (APP), and the normal metabolite of APP has been reported to have nerve cell protection function. Recent studies using genetic engineering have shown that toxic proteins such as β-amyloid accumulate in cells and blood vessels and become toxic to nerve cells, resulting in impaired brain function

To date, medication for the treatment of dementia has been used to treat acetylcholine precursors or to increase acetylcholine levels in the brain by administering a drug that inhibits acetylcholine degradation. Typical drugs include tacrine, donepezil, rivastigmine, and galantamine.

However, the existing dementia treatment agent mainly has only acetylcholinesterase inhibiting effect, but has no inhibitory effect on the formation of beta-amyloid. Therefore, it was impossible to cure dementia.

In addition, existing dementia therapies including various chemical substances have a problem that side effects such as nausea, vomiting, dizziness, diarrhea and nervous system disorders frequently occur.

Therefore, there is an urgent need to develop a therapeutic agent for preventing and treating dementia of natural origin, which has an effect of inhibiting beta-amyloid aggregates and has less side effects.

Korean Patent Publication No. 10-2010-0082090 Korean Patent Publication No. 10-2010-0012927

It is an object of the present invention to provide a pharmaceutical composition for prevention and treatment of degenerative brain diseases comprising an extract of a spectator.

Alzheimer's Disease (Alzheimer's Disease) is a type of dementia that gradually declines in mental function due to loss of function of brain tissue. It is the most common cause of dementia, and it is a degenerative brain disease that occurs mainly in elderly people over 65 years old. Alzheimer's disease progressively has symptoms of memory loss and cognitive impairment. Initially, learning ability and ability to remember recent events are severely reduced, but memories of old past like youth are maintained until disease progresses. In addition, there are serious injuries such as forgetting promises, misplaced things, getting lost in a familiar place, and failing to make money. In the mid-term, patients gradually feel difficulty in speaking and become unable to use the product well. As the dementia progresses, they are wandering the streets, becoming belligerent, and exhibiting urinary incontinence. Also, at the end, you lose the ability to eat, walk and communicate. Therefore, most patients die from vaginal infection within 3 to 20 years after the onset.

The pathological specifics of Alzheimer's disease include neurofibrillary tangles in which nerve fibers are entangled around the nucleus of nerve cells and senile plaques accumulated outside of nerve cells. Neurofibrillary tangles are aggregates produced by over-phosphorylation of tau protein, a protein that stabilizes microtubule-related proteins, the skeletal structure of nerve cells. As a result, the structure of normal microtubules is destroyed and livestock transport can not be performed. As a bundle of nerve fibers is generated in nerve cells, an increasing number of nerve cells become dysfunctional, and furthermore, die. Senile plaques are characterized by aggregation of glial cells, microglia, and astrocytic inflammatory cells around aggregated beta-amyloid (Aβ), the main component of which is beta-amyloid, which causes neurodegenerative toxicities It is known as a substance.

Beta-amyloid (Aβ) is a metabolite produced by cleavage of N-terminal and C-terminal by a protease from a type 1 integral membrane glycoprotein called APP (amyloid precusor protein) and consists of extracellular domain and membrane domain It is 36-43 peptides. The enzyme involved in β-amyloid production is β-secretase, which is called β-site APP-cleaving enzyme (BACE1), and gamma-secretase consisting of at least four protein complexes (presenilin 1 or 2, presenilin enhancer 2, anterior pharynx, nicastrin) -secretase, and amyloidogenic pathway is the metabolic process in which beta amyloid is produced by such a protease. APP cleaved by β-secretase (BACE1) is divided into an N-terminal domain called sAPPβ and a cytoplasmic domain called CTFβ (C99) at about 90 kDa. The resulting sAPPβ is secreted out of the cell. C99 is again secreted into the y-secretase To produce 4 kDa of beta-amyloid. In contrast, the non-amyloidogenic pathway is caused by the α-secretase that cleaves the α-site of APP and is a metabolic process that cleaves between the 16th and 17th amino acids and prevents formation of beta amyloid. α-secretase is a protease belonging to the α-disintegrin metalloproteinase domain (ADAM) family with disintegrin domain and metalloproteinase domain. ADAM-9, ADAM-10 and ADAM-17 are considered to be members of α-secretase. When α-secretase (ADAM10 or 17) is cleaved by APP, it is cleaved into a N-terminal domain called sAPPα and a cytoplasmic domain called CTFα (C83). C83 is again cleaved by γ-secretase to form a 3 kDa The piece is made.

In summary, the step of inducing neuronal apoptosis by beta-amyloid is firstly beta-amyloid is generated stepwise by amyloidogenic pathway from APP, and the resulting monomer-form of beta-amyloid coagulates to form an oligomer, It has recently been accepted that beta-amyloid in the form of beta-amyloid induces neuronal apoptosis. Therefore, the AD therapeutic agent that protects nerve cells by inhibiting beta-amyloid-induced neuronal cell death should have a function of inhibiting beta-amyloid production, inhibiting beta-amyloid aggregation, or beta-amyloid toxicity. Accordingly, the inventors of the present invention have conducted studies to find a natural extract having a function of inhibiting the production of beta-amyloid in order to develop a therapeutic and preventive material for degenerative brain diseases, particularly Alzheimer's

Accordingly, the present invention provides a pharmaceutical composition for the prevention and treatment of degenerative brain diseases comprising an extract of a spectator as an active ingredient.

The pharmaceutical composition for the prevention and treatment of degenerative brain diseases comprising the extract of the present invention as an active ingredient contains the extract extracted from a natural plant as an active ingredient and has an effect of inhibiting the formation of beta-amyloid aggregates. Therefore, And the like can be prevented and treated. In addition, since the extract derived from a natural product is an effective ingredient thereof, there is no problem about side effects or cytotoxicity.

FIG. 1 is a view showing extraction and fractionation processes of an extract of a spectator according to the present invention. FIG.
FIG. 2 is a diagram showing cell lines overexpressing amyloid-beta protein by immunofluorescence and Western blotting.
FIG. 3 is a chart for measuring cytotoxicity of a methanol extract of a crowd according to the present invention. FIG.
FIG. 4 is a chart for measuring cytotoxicity of a fraction extract of a crowd according to the present invention. FIG.
FIG. 5 is a graph showing the inhibitory effect of the methanol extract of a crowd according to the present invention on the formation of beta-amyloid aggregation by ELISA.
FIG. 6 is a graph showing the inhibitory effect of the fraction extract of a crowd according to the present invention on the formation of beta-amyloid aggregation by ELISA.
FIG. 7 is a graph showing the effect of the butanol fraction extract of the present invention on the sAPPβ and BACE1 (β-secretase) measured by Western blotting.
FIG. 8 is a diagram showing the result of measurement by Western blotting method in which the effect of the butanol fraction extract of the present invention on the sAPPβ of the present invention was measured.
FIG. 9 is a graphical representation of the result of measurement by Western blotting, in which the effect of the butanol fraction extract of the present invention on the BACE1 (β-secretase) of the present invention was measured.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a pharmaceutical composition for the prevention and treatment of degenerative brain diseases comprising an extract of a spectator as an active ingredient.

The crowd (Crassirhizomae Rhizoma) refers to the dry root of the rootstock and petiole of Dryopteris crassirhizoma Nakai, a perennial herb of the Aspidiaceae. The young leaves are used for edible purposes, and the roots are used as medicines in one room. It grows in shady mountains all over the country, and is distributed in Japan, Sakhalin, Kuril and Manchuria. The crowns are slightly bent and conical in shape with a length of 7-10cm and a diameter of 3-6cm. Around the roots are covered with roots, and the roots are gathered and bent in the shape of several pillar-shaped pillars. The surface is dark brown with brownish scaly leaves, the cross section of the rootstock is polygonal, and the base tissue is green with porous nature of the sea surface. The crowd shows antiparasitic, antipyretic, detoxifying, cold virus inhibiting, antibacterial and anticancer activities, and the water that is crowded is known to suppress the skin fungus. It has also been reported that the crowd exhibits uterine stimulation, hemostatic, analgesic, and anti-inflammatory effects. However, despite the recent research by the crowd, no studies on the prevention and treatment of degenerative brain diseases have been reported.

The degenerative brain disease as referred to in the present invention includes all those related to brain diseases caused by aging, preferably means Alzheimer's disease or dementia.

The tube extract according to the present invention may be one prepared by a conventional method for producing a plant extract. Specifically, it may be a cold extraction method, a warm extraction method, a heat extraction method, an ultrasonic extraction method, Can be used.

The extraction solvent for extracting the spectral extract according to the present invention may be at least one selected from the group consisting of water and an organic solvent. (See Fig. 1)

The organic solvent may be an alcohol having 1 to 5 carbon atoms or a dilute aqueous solution thereof, a polar solvent such as ethyl acetate or acetone, or a nonpolar solvent such as ether, chloroform, benzene, hexane or dichloromethane or a mixture thereof Methanol, more preferably 50% (v / v) to 99% (v / v) methanol.

The diluted water of the above-mentioned alcohol means that 1 to 99% (v / v) of alcohol is diluted in water.

The extraction temperature of the extract may be performed at 4 ° C to 120 ° C, preferably at room temperature. The extraction time may be from 12 hours to 120 hours. If the extraction temperature is lower than 4 ° C, the extraction efficiency may decrease. If the extraction temperature is higher than 120 ° C, the extract may be denatured. If the extraction time is less than 12 hours, the effective component may not be extracted partially, and if the extraction time exceeds 120 hours, impurities other than the active component may be extracted together.

The tubular extract can be extracted with an extraction solvent, and then filtered to remove filter residue. The filtrate can be concentrated using a vacuum rotary evaporator or a vacuum dryer, and then stored at room temperature.

In one embodiment of the present invention, the inhibition of beta-amyloid aggregation on the methanol extract of the crowd was measured, and it was found that beta-amyloid aggregation was inhibited in a concentration-dependent manner. (See FIG. 5) Accordingly, the pharmaceutical composition comprising the spectral extract of the present invention can be used as an agent for preventing or treating a degenerative brain disease caused by the formation (aggregation) of beta-amyloid in brain tissue.

In addition, the methanol extract of the above-mentioned spectra can be subjected to simultaneous or sequential fractionation using one or more solvents selected from the group consisting of hexane, chloroform, ethyl acetate, and butanol. (See Fig. 1)

The fractionation temperature may be performed at 4 캜 to 120 캜.

The fraction extract obtained by performing the fractionation process can be concentrated using a vacuum rotary condenser or a vacuum dryer, and then stored at room temperature.

In one embodiment of the present invention, the inhibitory activity on beta-amyloid production of the extracts of the spectra was measured. As a result, the butanol fraction extract (CRE-BuOH), which was sequentially fractionated with a solvent of hexane, chloroform, ethyl acetate and butanol, Indicating that amyloid production is inhibited. (See Fig. 6)

The pharmaceutical composition for the prevention and treatment of degenerative brain diseases comprising the extract of the present invention as an active ingredient according to the present invention can be applied to animals including humans.

The dosage of the pharmaceutical composition for the prevention and treatment of degenerative brain diseases can be determined in consideration of the administration method, the age and sex of the recipient, the severity of the patient, the state, the degree of absorption of the active ingredient in the body, the inactivation rate, , 0.1 mg / kg (body weight) to 500 mg / kg (body weight), 0.1 mg / kg (body weight) to 400 mg / kg (body weight) or 1 mg / 300 mg / kg (body weight), and may be administered once or several times in divided doses, but is not limited thereto.

The spectral extract according to the present invention may be used alone in the pharmaceutical composition for the prevention and treatment of degenerative brain diseases, and may further include a pharmaceutically acceptable carrier.

In this case, the content of the tubular extract in the pharmaceutical composition for the prevention and treatment of degenerative brain diseases may be 0.001% by weight to 99.9% by weight, 0.1% by weight to 99% by weight or 1% by weight to 50% And the content of the compound may be suitably adjusted depending on the intended use of the composition and the method of use.

Examples of the pharmaceutically acceptable carrier include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose , Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium But are not limited to, one or more selected from the group consisting of water, carbon dioxide, propylene glycol, liquid paraffin, and physiological saline. Any conventional pharmaceutically acceptable carrier may be used.

In addition, the composition for preventing or treating dementia including a spectral extract may be prepared by using a conventional filler, an extender, a binder, a disintegrant, an anticoagulant, a lubricant, a wetting agent, a pH adjuster, a nutrient, a vitamin, an electrolyte, A salt thereof, a protective colloid, a glycerin, a flavoring, an emulsifying agent or an antiseptic, and the like.

In addition, the pharmaceutical composition for the prevention and treatment of degenerative brain diseases of the present invention may contain substances known to those skilled in the art, such as tacrine, donepezil, Rivastigmine, galantamine, and the like may be additionally included.

When the spectral extract according to the present invention is used as a medicine, the administration method includes both oral and parenteral administration. In addition, the formulation of the pharmaceutical composition for the prevention and treatment of degenerative brain diseases may be varied depending on the method of use, and may be varied depending on the technique of the present invention to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal Lt; / RTI > may be formulated using methods well known in the art.

Solid formulations for oral administration include tablets, soft or hard capsules, pills, powders, suspensions, syrups, injections and granules, which may contain one or more excipients such as starch, calcium carbonate, Lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Forms for parenteral administration may be in the form of creams, lotions, ointments, alerts, liquids, aerosols, fluid extracts, elixirs, infusions, sachets, patches or injections.

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Example  1. Preparation of spectral methanol extract and fraction extract

1. Preparation of spectral methanol extract

1.2 kg of the dried blood was extracted with 90% methanol at room temperature for 3 days, filtered through filter paper (Nalgene, New York, NY, USA), and the filtrate was concentrated using a vacuum rotary evaporator (EYELA, Tokyo, Japan) . This procedure was repeated 3 times and the concentrate was stored at 4 ° C until use. The methanol in the spectra was in the form of an extracted brown crystalline powder, and a total of 65 g of the extract was obtained. (See Fig. 1)

2. Preparation of extracts of butanol fraction from crowd

The spectral extracts were fractionated with two solvents that did not mix with polarity differences. The methanol extracts obtained in the above step 1 were fractionated into hexane, chloroform, ethyl acetate, butanol and water, and the procedure was repeated three times. The concentrate was stored at 4 ° C until use. 3.2 g of the hexane layer, 0.6 g of the chloroform layer, 10.8 g of the ethyl acetate layer, 14 g of the butanol layer and 28 g of the water layer were obtained, respectively. (See Fig. 1)

Experimental Example  1. Assessment of cytotoxicity of spectral extracts

1. Production of transfected cell lines overexpressing amyloid-beta precursor protein (APP)

The CHO (chinese hamster ovary) cells used in the experiments were purchased from the Korean Cell Line Bank (KCLB; Korea). The cells were cultured in RPMI medium containing 10% FBS at 37 ° C in a 5% CO ₂ incubator. The medium was changed every 3 days.

(Reverse, SEQ ID NO: 2) to 5'-ctg gcc tgc tgg ctg aac ccc-3 '(forward, SEQ ID NO: 1), 5'-gtc cac gta tct ttg ggt ctt- primer was used to amplify only the APP portion, inserted into the pcDNA 3.1 vector, and the prepared APP-pcDNA 3.1 vector was transfected into the cultured CHO cells. For the transfection injection according to Lipofectamine®2000 Transfection Reagent protocol in ^{60mm dish CHO cell 1.2 X 10 6} / 5ml one of seeding the cells, then after 24 hours into the APP-pcDNA3.1 320ng / Lipofectamine 0.8 μl 24 hours at 37 ℃ Lt; / RTI > To select only the transfected cells (APP _wt -CHO) in the transfected CHO cells, the cells were cultured in medium containing 2.5 mg / ml of geneticin once every 3-4 days.

2. Selection of APP _wt -CHO cell line by immunofluorescence

A cover glass was inserted into each well of a 24-well plate, and 2 × 10 ⁵ APP _wt -CHO cells were seeded on the cover glass. After incubation for 24 hours, 4% paraformaldehyde was added to 200 μl / well for 30 minutes at room temperature . After blocking with 5% BSA (in PBS) for 1 hour, the primary antibody, APP / β-amyloid (NAB228) Mouse mAb (Cell signaling) was diluted (1:50) with 3% BSA And reacted at 4 DEG C for 12 hours. After washing with PBS, 20 μl of a fluorochrome-conjugated secondary antibody was diluted (1:50) with Alexa Fluor 555 donkey anti-mouse IgG (H + L) -invitrogen 3% BSA and incubated at room temperature for 1 hour. The nuclei were stained with DAPI and the degree of transfection was observed through a fluorescence microscope.

3. Evaluation of cytotoxicity of spectral extracts

MTT-based cytotoxicity assay was performed to examine the cytotoxicity of the extracts of the spectra of APP _wt -CHO cells. The cells treated with the drug were cultured for 24 hours, and then MTT solution was added at a concentration of 5 mg / ml for 3 hours. After removing the medium and dissolving in DMSO for 30 minutes, the absorbance was measured at 540 nm using an Emax precision microplate reader (Molecular Devices, CA, USA).

The cytotoxicity of the five methanol extracts (CRE-MeOH) from 2, 10, 50, and 100 μg / ml was not toxic, ml, 10 μg / ml in the chloroform layer, 10 μg / ml in the ethyl acetate layer, 10 μg / ml in the butanol layer, and 10 μg / ml in the water layer. (See Figures 3 and 4)

Experimental Example  2. ELISA Evaluation of Efficacy on the Generation of Beta-Amyloid by the Spectral Extract

The APP _wt -CHO cells constructed in step 1 of Experimental Example 1 were cultured in a 96-well plate (6 x 10 ⁴ cells / well) for 4 hours, and the methanol extracts (2, 10, 50, 100 ug / ml) And 50 ug / ml hexane / ethyl acetate / butanol / water fraction extracts were cultured and cultured for 24 hours. After incubation, the supernatant was transferred to an EP tube and centrifuged at 3000 xg for 5 minutes to remove cell debris. The amount of beta-amyloid in the supernatant was measured using a Human A [beta] 40 ELISA kit (Invitrogen KHB3482). Briefly, 10 μl of sample supernatant, 40 μl of dilution solution, 50 μl of Human Aβ40 Detection antibody solution, and AEBSF (1 mM) were added to each well and reacted at room temperature for 3 hours. After removing the reaction solution, 100 μl of washing solution was added and washed 4 times. 100 μl of anti-rabbit IgG HRP working solution was added to each well and reacted at room temperature for 30 minutes. After removing the reaction solution, 100 μl of washing solution was added and washed 5 times. Then, 100 μl of Stabilized Chromogen (TMB solution) was added to each well and reacted in a dark room for 30 minutes. After that, 100 μl of stop solution was added to each well, and the amount of β-amyloid was measured at 450 nm wavelength using a microtiter plate reader.

As a result, the amount of beta-amyloid decreased to 74% of the control group and 27% of 100 μg / ml of the methanol extract of CRE-MeOH (CRE-MeOH) Butanol and water layer showed 60% level of the butanol layer and 58% level of water layer, respectively, in the hexane, ethyl acetate, butanol, and water layers. (See Figures 5 and 6)

Experimental Example  3. Western blot 분석 Of spectral extracts sAPP beta and BACE1 level  Evaluation of efficacy on change

The cells were incubated with 10, 50, and 100 μg / ml of the extracts in a 60-mm dish for 24 hours, washed with PBS, incubated with trypsin for 3 minutes, removed, and transferred to an EP tube. Cell pellet was prepared by centrifugation at 1000 x g, supernatant was removed, and 50 μl of lysis buffer was added. Proteins were extracted by dissolving at -20 ° C for 30 min and quantitated using SMART ™ BCA kit (Intron, Korea). Sample loading buffer was added to the extracted protein, boiled in boiling water at 100 ° C for 10 minutes, and stored at -20 ° C until use. Proteins were separated by electrophoresis with 7.5% gel or 15% acrylamide gel and transferred to PVDF membrane. In order to prevent nonspecific binding of the PVDF membrane to the antibody, the cells were incubated in blocking buffer (5% skin milk in PBS) for 1 hour, washed with PBS solution (0.1% PBST) The primary antibody to the test protein was added and reacted at 4 ° C for 12 hours. BACE1 antibody (1: 1000, Abcam), APP c-terminal antibody (1: 1000, Sigma A8717) was used for the primary antibody used. , sAPPβ antibody (1: 1000, IBL 18957) and beta-actin antibody (1: 1000, BD biosciences, CA, USA). PBST solution for 15 minutes, and reacted with a secondary antibody (5% skin milk in PBS) for 1 hour. Proteins were then identified on the G: Box iChemiXT Imager (Syngene, Cambridge, UK) after ECL-spray (Advanta, CA, USA).

As a result, there was no significant change in the expression level of sAPPβ compared to the control group. However, 50 μg / ml was decreased to 20% in the control group and 8 μg / ml in 100 μg / ml, 100, and 50%, 30%, and 8% of the control, respectively. (See Figs. 7 to 9)

The preparation examples described below are for the purpose of simple reference only and do not limit the scope of the present invention. The content of the constituent components described in the following Preparation Examples may be varied according to the intensity of inflammation, the purpose and the circumstances, and may be modified according to a conventional method for producing a pharmaceutical composition.

Manufacturing example  One. Sanje  Produce

The components listed in Table 1 below were mixed and packed in airtight bags to prepare powders.

Spectral butanol fraction extract 100 mg Lactose 300 mg Talc 10 mg

Manufacturing example  2. Preparation of tablets

The ingredients listed in Table 2 were mixed and tableted according to a conventional tablet preparation method.

Spectral butanol fraction extract 10 mg Lactose 100 mg Corn starch 100 mg Magnesium stearate 2 mg

Manufacturing example  3. Preparation of capsules

The ingredients listed in Table 3 below were mixed and filled into gelatin capsules.

Spectral butanol fraction extract 10 mg Crystalline cellulose 3 mg Lactose 14.8 mg Magnesium stearate 0.2 mg

Manufacturing example  4. Preparation of injections

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

(After filtration for main use), butanol fraction extract 10 mg Mannitol 200 mg Na ₂ HPO _{4 12} H ₂ O 30 mg

<110> Dankook University Cheonan Campus Industry Academic Cooperation Foundation <120> Pharmaceutical Composition Comprising Dryopteris crassirhizoma          Extract for the Treatment of Degenerative Brain Diseases <130> HY131227 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 1 ctggcctgct ggctgaaccc c 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 2 gtccacgtat ctttgggtct t 21

Claims (7)

A pharmaceutical composition for prevention and treatment of degenerative brain diseases comprising an extract of a spectator as an active ingredient,
The above-mentioned spectral extract was obtained by extracting the spectra from 50% (v / v) to 99% (v / v) methanol,
Wherein the extract is a butanol fraction extract obtained by successively fractionating with hexane, chloroform, ethyl acetate, and butanol. The method according to claim 1,
Wherein the degenerative brain disease is Alzheimer's disease or dementia. delete delete delete The method according to claim 1,
Wherein the pharmaceutical composition is formulated into a formulation selected from the group consisting of tablets, soft or hard capsules, pills, powders, suspensions, syrups, injections and granules. The method according to claim 1,
Wherein the pharmaceutical composition is for oral or parenteral administration.

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