KR20200029702A - Pharmaceutical composition including abeliophyllum distichum extract for preventing or treating liver injury - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating dementia containing an Abeliophyllum distichum extract as an active component. The present invention is to provide the pharmaceutical composition for preventing or treating dementia containing the Abeliophyllum distichum extract as the active component, as novel efficacy of the Abeliophyllum distichum extract. The pharmaceutical composition containing the Abeliophyllum distichum extract can be provided for stable and effective dementia prevention or treatment.

Description

Pharmaceutical composition for the prevention or treatment of dementia, which includes the extract of the hedgehog tree as an active ingredient {PHARMACEUTICAL COMPOSITION INCLUDING ABELIOPHYLLUM DISTICHUM EXTRACT FOR PREVENTING OR TREATING LIVER INJURY}

The present invention relates to a pharmaceutical composition for the prevention or treatment of dementia, which includes the extract of the hedgehog tree as an active ingredient.

The Abeliophyllum distichum is a tree plant belonging to the family Oleaceae, and is a species of genus 1 and unique to Korea. Designated as a natural monument in Korea, it is protected in native areas of certain regions and is mainly distributed in Jincheon, Goesan-gun, and Yeongdong, Chungcheongbuk-do. Studies on the extract of hedgenut tree began to be actively conducted relatively recently, and it has been reported that glycosides extracted from hedgehog leaf inhibit angiotensin converting enzyme that regulates blood pressure (Oh H, Kang) DG, Kwon TO, Jang KK, Chai KY, Yun YG, Chung HT, Lee HS, Phytotherapy Research 17: 811-813, 2003). Bar (Kor. J. Herbology 2011; 26 (4): 95-99). In addition, Korean Patent Registration No. 10-706131 and Korean Patent Registration No. 10-656287 disclose use for anti-cancer agents and anti-inflammatory agents, including stalk extract. In addition, Korean Patent Publication No. 2016-0089257 (published on July 27, 2016) describes the antioxidant effect, and Korean Patent Publication No. 2016-0089256 (published on July 27, 2016) and Korean Patent Publication No. 2016-0114801 ( 2016.10.06.Public) describes the atopy improvement effect and the odor removal effect of the stalk extract.

The present inventors tried to reveal the functionality of the stalk extract, and as a result, confirmed that the stalk extract has a variety of functions in addition to the previously known functionality, and completed the present invention.

Korean Patent Registration No. 10-0706131 Korean Patent Registration No. 10-0656287 Korean Patent Publication No. 2016-0089257 Korean Patent Publication No. 2016-0089256 Korean Patent Publication No. 2016-0114801

 Oh H, Kang D-G, Kwon T-O, Jang K K, Chai K- Y, Yun Y-G, Chung H-T, Lee H-S, Phytotherapy Research 17: 811-813, 2003. Jaeho Park, Kor. J. Herbology 2011; 26 (4): 95-99.

The present invention is to provide a pharmaceutical composition for preventing or treating dementia, which includes as an active ingredient, as a new efficacy of the extract of the pear tree extract.

The pharmaceutical composition for preventing or treating dementia of the present invention for achieving the above object includes an extract of the cactus tree as an active ingredient. The coleoptera extract contained in the pharmaceutical composition defined in the present invention includes not only coleoptera extract, but also a solvent fraction thereof or a mixture thereof.

In one embodiment, the coleoptera extract may be coleoptera extract with water, a lower alcohol having 1 to 6 carbon atoms, or a mixed solvent thereof.

In one embodiment, the coleoptera extract may be a coleoptera extract with 30% to 95% ethanol or a hot water extract.

In one embodiment, the coleoptera extract may be a coleoptera extract or hot water extract by 70% ethanol.

In one embodiment, the coleoptera extract fractionates coleoptera extract by 30% to 95% ethanol with at least any one of acetate and butanol from coleopteran extract by water, lower alcohol having 1 to 6 carbon atoms, or a mixed solvent thereof. It can be a fraction.

In one embodiment, the coleopteran extract shows the ability to inhibit beta amyloid aggregation or inflammation of the brain microglia, and to inhibit vascular dementia.

In one embodiment, the coleoptera extract may be an extract of the leaves or stems of coleoptera.

In the pharmaceutical composition for preventing or treating dementia comprising the above-mentioned saponica extract as an active ingredient, it is confirmed that the saponica extract, which is an active ingredient, exhibits an inhibitory effect on beta amyloid aggregation or inflammation of brain microglia, and inhibits vascular dementia. Did. The pharmaceutical composition comprising the extract of the hedgehog tree may be provided for stable and effective dementia prevention or treatment.

1 is a view showing the results of evaluating cell viability for showing the effect of inhibiting brain cell inflammation of 70% alcoholic beverage extract and stem hot water extract according to an embodiment of the present invention.
2 is a view showing the results of the NO secretion inhibitory ability evaluation to show the effect of inhibiting brain cell inflammation of the 70% ethanol extract and stalk stem water extract according to an embodiment of the present invention.
Figure 3 is a view showing the suppression of inflammatory cytokine secretion for showing the brain cell inflammation inhibitory effect of 70% alcoholic extract of the myrtle leaf and the myrtle stem hot water extract according to an embodiment of the present invention.
4 is a view for explaining the change in the number of cranial nerve cells in the brain (hippocampus) of the vascular dementia-induced animal model according to administration of 70% alcoholic extract of the myrtle leaf and hot water extract of the myrtle stalk according to an embodiment of the present invention.

In the present invention, the stalk tree for obtaining a stalk tree extract may include one obtained from any one or two or more parts of leaves, stems, fruits, roots, husks, and seeds in Korea, and preferably stalk tree leaf extracts Or a stem extract is used. In addition, preferably, the leaf extract and the stem extract may be a leaf / stem harvested during a specific time period or a specific period during a period between spring and autumn, and a leaf / stem harvested during a period between spring and autumn. It may be used. In the present invention, extraction of the active ingredient from the stalk tree can be carried out by a commonly known solvent extraction method. In the present invention, preferably, it may be a stalk leaf extract or a stalk leaf water extract.

In order to obtain a cactus extract, a cactus is collected and cut, washed, dried and crushed to a desired site, for example, cactus leaves and / or stems; 3 to 30 times, preferably 10 to 20 times water or lower alcohol (e.g., methanol, ethanol, butanol, etc.), or a mixed solvent thereof, extracted at 60 ° C. to 75 ° C. The extraction process is performed for at least 1 hour at a temperature. At this time, the extraction of alcohol using lower alcohol may be 30% alcohol to 95% alcohol. If it is less than 30% alcohol, it is preferable to perform at 30% to 95% alcohol because more practically no alcohol soluble active ingredients can be obtained, and when it is more than 95% alcohol, there are few water-soluble active ingredients, more preferably May be 30% to 70% alcohol, and most preferably 70% alcohol. At this time, the extract can be extracted until the concentration of the soluble solids exceeds 0 brix but reaches 30 brix. In one example, the extract may be extracted until the concentration of soluble solids reaches 20 to 21 brix. At this time, as a mixed solvent, the mixing ratio of water and lower alcohol may be 1: 1 to 1:10. As the extraction method, hot water extraction, cold immersion extraction, reflux cooling extraction, ultrasonic extraction, steam distillation extraction, or the like may be performed one to five times in succession. After extraction, it is filtered at a temperature condition of 40 ° C to 60 ° C, the filtrate is concentrated under reduced pressure, and then dried by a freeze dryer to obtain a stalk extract.

After suspending the extract of the stalk tree obtained in this way, the system is separated using an extraction solvent, and concentrated under reduced pressure to obtain a fraction of each extract solvent of the stalk extract. As an example of a solvent used to obtain a cactus extract, purified water or an organic solvent having 1 to 6 carbon atoms can be used. Examples of the organic solvent, ethanol, methanol, propanol, butanol, glycerin, ethyl acetate, ethyl acetate, butylene glycol, propylene glycol (propylene glycol) glycol), dichloromethane, chloroform, ethyl ether, hexane, and the like. These may be used alone or in combination of two or more.

In one embodiment, the hexane, dichloromethane, ethyl acetate, water-saturated butanol, and water may be used to obtain a fraction of the extract of the coleopteran extract, thereby hexane layer, dichloromethane layer, ethyl acetate of the coleopteran extract Fractions of layers, butanol layers and water layers can be obtained. That is, hexane treatment can be performed to obtain a hexane-soluble fraction and a water-soluble fraction, and again, the water-soluble fraction can be extracted with dichloromethane to obtain a water-soluble fraction and a dichloromethane-soluble fraction, ethyl in the water-soluble fraction Acetate can be added to obtain an ethyl acetate soluble fraction and a water soluble fraction, and the water soluble fraction can be extracted with butanol to obtain a butanol soluble fraction and a water soluble fraction.

As an active ingredient of the pharmaceutical composition according to the present invention, as a result of evaluating the inhibitory effect of beta amyloid aggregation and the inhibitory effect of inflammation of BV-2 brain microglial cells in order to confirm the efficacy of the extract of the hedgehog tree as a pharmaceutical composition for preventing or treating dementia, It was confirmed that the extract of the cactus tree of the present invention has an effect of inhibiting beta amyloid aggregation. In addition, since it has no cytotoxicity and has little toxicity and side effects as a natural substance, it can be used safely for a certain period of time to prevent or treat dementia, and it has been confirmed that it inhibits the secretion of NO, an inflammatory marker, and also has cytokine suppression ability. Was confirmed. Through the efficacy of such a hedgehog extract, a hedgehog extract may be provided as a pharmaceutical composition for preventing or treating dementia.

The pharmaceutical composition according to the present invention, as an active ingredient of the hedgehog tree extract, may include a pharmaceutically acceptable excipient.

The pharmaceutical composition according to the present invention, as an active ingredient of the hedgehog tree extract, may contain one or more known active ingredients having a dementia prevention or treatment effect.

The composition of the present invention may be prepared by including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration. Pharmaceutically acceptable carriers can be used by mixing one or more of these components: saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and antioxidants, buffers if necessary. , Other conventional additives such as bacteriostatic agents can be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets.

The active ingredient in the entire pharmaceutical composition may be included as an active ingredient in the extract of the stalk tree according to the present invention in an amount of 0.001 to 90% by weight, preferably 0.01 to 15% by weight, and more preferably 0.01 to 10% by weight. The dosage amount of the pharmaceutical composition thus prepared can be appropriately adjusted according to the individual difference formulation such as age and degree of lesion, and the daily dosage for an adult can be used in an effective amount of 0.01 mg to 10,000 mg. There is, for example, the daily dosage may be from 0.01 mg to 1,000 mg, it is preferable to be divided into once to several times a day.

The pharmaceutical composition of the present invention may further contain a preservative, a stabilizer, a hydrating or emulsifying accelerator, a salt or buffer for controlling osmotic pressure, a disintegrant, a binder, a lubricant, a diluent, and other therapeutically useful substances. It can be formulated in various oral or parenteral dosage forms according to conventional methods.

Formulations for oral administration include, for example, tablets, pills, hard and soft capsules, liquids, suspensions, emulsifiers, syrups, granules, etc. These formulations, in addition to active ingredients, diluents such as lactose, dextrose, sucrose , Mannitol, sorbitol, cellulose and glycine, lubricants such as silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycol. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, and optionally starch, agar, alginic acid or sodium salts thereof It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweeteners. Tablets can be prepared by conventional mixing, granulating or coating methods. In addition, a representative formulation for parenteral administration is an injectable formulation, preferably an isotonic aqueous solution or suspension.

However, it should be understood that the actual dosage of the active ingredient should be determined in light of a number of related factors such as the severity of the symptoms, the route of administration chosen, the age, sex, weight and health of the subject.

Dosages suitable as pharmaceutical compositions according to the present invention are usually readily available to skilled physicians to determine and prescribe effective doses for the skin. The daily dosage of the drug according to the present invention varies depending on various factors such as the degree of obesity progression, the onset time, age, health condition, complications, etc. of the subject to be administered, but is generally the above-mentioned weight ratio when referring to adults. Compositions combined with 1 to 500 mg / kg, preferably 30 to 200 mg / kg can be administered in divided portions 1 to 2 times a day, and the dosage is to limit the scope of the present invention in any way. no.

Hereinafter, embodiments according to the present invention will be described in detail. However, the following examples are provided only for easier understanding of the present invention, and the present invention is not limited by the examples.

Below, the reagents used for the preparation and experiment of the extract were purchased from Sigma Aldrich and used as they are.

Example: Preparation of stalk extract

After crushing and crushing 2 kg of Misun tree leaves native to Goesan, Chungbuk, add 40 L of 30%, 70%, and 95% ethanol, extract for 4 hours at 75 ° C, filter with filter paper, and extract under reduced pressure at 50 ° C. Was concentrated to 2L. The concentrated extract was dried with a lyophilizer to obtain a leaf of the hawthorn (30% alcohol (30% ethanol): 0.89 kg, 70% alcohol: 1.02 kg, 95% alcohol: 0.48 kg).

In addition, 40 L of distilled water was added to 2 kg of the stalk of the stalk and extracted for 4 hours at 100 ° C., filtered with filter paper, and then the extract was concentrated to 2 L under reduced pressure at 50 ° C. The concentrated extract was dried with a lyophilizer to obtain a hot water extract (0.75 kg) of stalk.

20 g of the 70% extract of saffron leaf extract obtained as above was suspended in 200 mL of distilled water, systemically separated with hexane, dichloromethane (methylene chloride), ethyl acetate, water-saturated butanol and water, and concentrated under reduced pressure to extract the saffron leaf extract. Fractions of hexane layer (0.4 g), dichloromethane layer (1.5 g), ethyl acetate layer (2.0 g), butanol layer (10.0 g), and water layer (6.1 g) were obtained. All extracts were used by dissolving the fractions obtained for cell experiments in dimethyl sulfoxide (DMSO) at a concentration of 100 mg / mL.

Experimental Example 1-1: Beta amyloid aggregation inhibition ability evaluation-1

A beta amyloid aggregation inhibitory capacity evaluation experiment was performed using the stalk leaf / stem extract (sake or hot water) obtained according to the above example. Fluorescence analysis (ThioflavinT assay) was used to measure the degree of aggregation of beta amyloid (β-amyloid, Aβ). The beta amyloid (Aβ1-40) peptide was stored at minus 80 ° C, and the final concentration used in the measurement was used at a concentration of 5 μM. The stalk extract (alcohol and hot water extract) obtained according to the present invention was dissolved in 99.5% DMSO immediately before the experiment and diluted by concentration with a buffer solution, and 5 μM of thioflavin T used as a fluorescent material was 0.01 It was prepared by dissolving in M sodium phosphate buffer solution.

To measure the degree of Aβ1-40 aggregation inhibition, 5 μl of fine extracts by fractions according to various solvent conditions were added to 92.5 μl of 0.01 M sodium phosphate buffer solution, and 2.5 μl of 5 μM Aβ1-40 was added and cultured at 37 ° C. for 26 hours. Was aggregated, and 2,000 μl of 5 μM thioflavin T was added and measured with a fluorescence spectrometer. At this time, the wavelength value of the fluorescence spectrometer used was Ex = 446 nm (slit width = 5 nm), Em = 490 nm (slit width = 10 nm), and the same experiment was repeated at least three times or more to represent the average value. The results are shown in Table 1 below.

Sample classification Sample Concentration
(μg / mL) β-amyloid
Inhibition of aggregation (%) Ab 0 0 Ab + 0.4% DMSO 0 0 Ab + ovanillin 100 47.7 Ab + 30% of fresh leaves 100 40.3 Ab + sake leaf alcohol 70% 100 73.3 Ab + sake leaf alcohol 95% 100 50.7 Ab + streak stem water 100 78.5

In Table 1, "Ab" represents beta amyloid, "Ab + 0.4% DMSO" represents a case where only DMSO is mixed with beta amyloid, "Ab + ovanillin" represents treatment of the positive control ovanillin, and "Ab + streak" "30% of leaf alcohol" refers to a sphere using an extract extracted from 30% alcohol, and "Ab + 70% leaf extract" represents a sphere using an extract extracted from 70% alcohol, and "Ab + "95%" refers to a sphere using an extract extracted from 95% sake tree, and "Ab + stalk stem water" refers to a sphere using an extract obtained by hot water extraction of the stem of a sap tree.

Referring to Table 1, among the treatment groups of the extracts of the stem stem extract and the leaves of the stem tree leaves (30% alcohol, 70% alcohol, 95% alcohol), the aggregation inhibition (%) of beta amyloid was the highest in the 70% alcohol extract treatment. You can see that the value is displayed. The higher the value of aggregation inhibition (%), the higher the aggregation inhibitory effect of Aβ1-40, and the lower the value, the lower the aggregation inhibition efficiency is. It can be seen that when the streak stem extract was used and when the 70% alcohol extract was used, the aggregation inhibitory efficacy of Aβ1-40 was higher than that of the 30% or 95% alcohol extract. Compared with the ovanillin treatment, it has a cohesive inhibitory effect of beta amyloid by the stalk extract according to the present invention, and in particular, the 70% alcoholic extract of the stalk leaf and the hot water extract of the stalk stem have higher aggregation inhibition than the ovanillin treatment Can be confirmed.

Experimental Example 1-2: Beta-amyloid aggregation inhibition ability evaluation-2

The beta amyloid aggregation inhibitory capacity evaluation experiment by fractions was performed using the stalk leaf / stem extract (sake or hot water) obtained according to the above example. The aggregation inhibition performance evaluation experiment was performed and analyzed substantially the same as that performed in Experimental Example 1-1. Table 2 shows the results.

Sample classification Sample Concentration
(μg / mL) β-amyloid
Aggregation suppression (%) Ab 0 0 Ab + 0.4% DMSO 0 0 Ab + ovanillin 100 47.7 Ab + streak stem water 100 78.5 Ab + Seaweed alcohol 70% 100 73.3 Ab + stray leaf hexane fraction 100 13.2 Ab + stray leaf methyl chloride fraction 100 49.5 Ab + stray leaf ethyl acetate fraction 100 14.4 Ab + stray leaf butanol fraction 100 15.3 Ab + Fresh leaf fraction 100 47.5

In Table 2, "Ab" represents beta amyloid, "Ab + 0.4% DMSO" represents a case where only DMSO is mixed with beta amyloid, "Ab + ovanillin" represents treatment of the positive control ovanillin, and "Ab + streak" Stem hydrothermal "refers to the case where an extract obtained by hot water extraction of the trunk of the stalk tree is used, and" Ab + sake leaf extract 70% "refers to the case where the extract obtained by extracting a stalk tree leaf by 70% alcohol is used," Ab + stalk leaf hexane fraction ", The "Ab + fine leaf methyl chloride fraction", "Ab + fine leaf ethyl acetate fraction", "Ab + fine leaf petanol fraction" and "Ab + fine leaf petite fraction" extract hexane, methyl chloride, ethyl after extracting 70% of each leaf The case where fractions each fractionated with acetate, butanol and water is used.

Referring to Table 2, among the treatments of extracts by 70% alcoholic extract of coleoptera leaf and 70% alcoholic extract of coleoptera leaf (hexane, methyl chloride, ethyl acetate, butanol, water) Aggregation inhibition was the highest, and it could be seen that the streak leaf methyl chloride fraction and the water fraction were similar or higher when compared to the positive control ovanillin.

According to a look at Table 1 and Table 2, it can be confirmed that the coagulation inhibitory effect of beta amyloid is obtained by the stalk extract according to the present invention. In particular, it can be seen that the 70% alcohol extract of the stalk tree leaf and the hot water extract of the stalk stem have a very high aggregation inhibitory effect of beta amyloid.

Experimental Example 2: Inflammation inhibitory effect of BV-2 brain microglia-cell survival rate

Cytotoxicity evaluation of streak extract was performed by observing cytotoxicity against BV-2 by treating streak extract by concentration on brain microglial cells (BV-2 microglial cells).

Specifically, after culturing the microglia in DMEM cell culture medium (Dulbecco's Modified Eagle's medium) containing 5% FBS at 37 ° C and 5% CO ₂ conditions, and spreading it on a 96 well plate to be 2 ㅧ 10 ⁴ cells per well. , The cells were completely attached while incubating for 12 hours, and LPS was treated with 50, 100, and 200 μg / mL treatments of the group and the group of treatments for 20 hours at the concentrations of 50, 100, and 200 μg / mL. (Lipopolysaccharide) was evaluated at a concentration of 1 μg / mL for the cell viability of the treated group for 24 hours in combination. Evaluation of cell viability was performed according to the MTT method, and the survival rate was calculated as% survival compared to the untreated group. The results are shown in FIG. 1.

1 is a view showing the results of evaluating cell viability for the brain cell inflammation inhibitory effect of 70% alcoholic beverage extract and stem hot water extract according to an embodiment of the present invention.

Referring to FIG. 1, the cell viability as a result of treatment by concentration (50, 100, 200 μg / mL) of the hedgehog leaf 70% alcoholic extract and the hedgehog stem hot water extract according to the treatment and no treatment conditions of the inflammation-inducing LPS Through this, it can be confirmed that there is no cytotoxicity to brain microglial cells (BV-2 microglial cells).

Experimental Example 3: Inhibitory effect of BV-2 brain microglial cells-NO inhibitory ability

After dispensing BV-2 cells into a 96 well plate at 2 ㅧ 10 ⁴ cells / well, cells are completely attached while incubating for 12 hours in a 37 ° C, 5% CO ₂ incubator. The cultured upper layer of the treatment group treated with 24 mL at a concentration of / mL and 50, 100, 200 μg / mL for the hedgehog tree extract and at the same time treated with LPS, an inflammatory substance, at a concentration of 1 μg / mL for 24 hours The liquid was separated. The content of NO was measured in the separated culture supernatant. The content of NO was treated with the same amount of Griess (Sigma) reagent in 100 μL of the separated culture supernatant, reacted for 10 minutes, and absorbance at 517 nm was measured using a microplate reader. The concentration of nitrogen oxide was calculated by comparison with a standard straight line obtained using sodium nitrite (NaNO ₂ , Sigma). The results are shown in FIG. 2.

2 is a view showing the results of the NO secretion inhibitory ability evaluation to show the effect of inhibiting brain cell inflammation of the 70% ethanol extract and stalk stem water extract according to an embodiment of the present invention.

Referring to Figure 2, when the treatment of the inflammation-inducing substance LPS, the secretion of NO was increased in the untreated tree extract, but 70% alcoholic extract of the tree leaves and the hot water extract of the tree leaves were used for brain microglial cells (BV-2 microglial cell). It can be seen that when the concentration (50, 100, 200 μg / mL) was treated by concentration, NO, an inflammation marker that is secreted by LPS, is decreased in a concentration-dependent manner. The treatment of the coleopteran extract reduces the secretion capacity of NO by the treatment of LPS, and it can be confirmed that it is effective in the inflammatory reaction.

Experimental Example 4: Inhibitory effect of BV-2 brain microglial cells-cytokine inhibitory ability

After dispensing BV-2 cells into a 96 well plate at 2 × 10 ⁴ cells / well, cells are completely adhered while incubating for 12 hours in a 37 ° C, 5% CO ₂ incubator. The cultured upper layer of the group treated with 24 mL at a concentration of / mL and 50, 100, and 200 μg / mL of the hedgehog tree extract and treated with LPS, an inflammatory substance, at a concentration of 1 μg / mL for 24 hours at the same time The liquid was separated. The content of inflammatory cytokines TNF-α and IL-6 in the separated culture supernatant was measured. The cytokine content was measured using an ELISA kit (eBioscience Co. (company name), San Diege, CA, USA), and the concentration of the cytokine was calculated from a standard curve calculated from a standard solution included in the kit. The results are shown in FIG. 3.

Figure 3 is a view showing the suppression of inflammatory cytokine secretion for showing the brain cell inflammation inhibitory effect of 70% alcoholic extract of the myrtle leaf and the myrtle stem hot water extract according to an embodiment of the present invention.

Referring to Figure 3, when the treatment of the inflammation-inducing substance LPS, secretion of inflammatory cytokines (TNF-α and IL-6) was increased in the non-treated tree extract, but 70% alcoholic extract of the tree leaf and hot water extract of the tree leaf TNF-α and IL-6, which are inflammation markers that are secreted by LPS when the cells are treated by concentration (50, 100, 200 μg / mL) in brain microglial cells (BV-2 microglial cells), are concentration-dependent. You can see that it decreases. It can be seen that the treatment of the coleopteran extract was effective in the inflammatory reaction, as it was observed to decrease the secretory ability of inflammatory cytokines by treatment of LPS.

Experimental Example 5: Efficacy of inhibiting vascular dementia

SD male rats of 8 weeks of age were administered at 10 mg / kg / day for 5 days of the extract of the hedgehog tree, and vascular dementia was induced on the 3rd day of administration. For the induction of vascular dementia, a model was used to limit the blood flow to the brain to less than 30% of the physiological blood flow by ligating both sides of the carotid artery for 10 minutes. 9 days after the end of the administration, cardiac perfusion was fixed with 4% paraformaldehyde to extract the brain from the experimental animals, and the extracted brain was fixed with the same solution for 24 hours. The washed brain was dehydrated and paraffinized through an alcohol series. It was cut into 7 um using a microtome, and a part with a hippocampus was selected and produced as a slide. Of these, 5 or more slides per horse were randomly selected for H-E staining, and the number of live neurons in the hippocampus CA1 region was expressed as a value. The results are shown in Fig. 4.

4 is a view for explaining the change in the number of cranial nerve cells in the brain (hippocampus) of the vascular dementia-induced animal model according to administration of 70% alcoholic extract of the myrtle leaf and hot water extract of the myrtle stalk according to an embodiment of the present invention.

In FIG. 4, “SHAM” refers to a gastric surgical instrument, “VD” refers to a vascular dementia inducer, and “VD + compound 1” refers to a group that is orally administered with 70% extract of unsold alcohol for 5 days to a vascular dementia inducer. "VD + compound 2" represents a group in which stalk stem hot water extract was orally administered for 5 days to a vascular dementia-inducing tool.

Referring to Figure 4, the number of living neurons in the gastric surgery was the most present, and in the case of vascular dementia inducers, the number of brain neurons was markedly reduced, and 70% alcoholic extract of the myelin leaf was administered and the hydrothermal stem hydrothermal extract It can be seen that the number of living cranial nerve cells in the dosing area increased higher than that of the vascular dementia inducing cells.

Therefore, it can be seen that the administration of the 70% alcoholic extract of the myrtle leaf and the hot water extract of the myrtle stem prevents the cell death of cranial nerve cells according to the induction of vascular dementia, thereby showing an inhibitory effect of vascular dementia.

The following is an example of a formulation for the pharmaceutical composition of the present invention.

Formulation Example 1: Preparation of pharmaceutical preparation

1. Preparation of powder

A powder was prepared by mixing 500 mg of coleopteran extract, 300 mg of lactose, 190 mg of corn starch and 10 mg of magnesium stearate, and filling the airtight bag according to a conventional powder manufacturing method. The final weight of the contents was 1,000 mg.

2. Preparation of tablets

According to a conventional tablet manufacturing method, 250 mg of hedgenut tree extract, 100 mg of lactose, 130 mg of microcrystalline cellulose, 15 mg of sodium starch glycolate and 5 mg of magnesium stearate were mixed and granulated and tableted with a tableting machine.

3. Preparation of Injection

According to the conventional injection preparation method, 10 mg of coleoptera extract, 180 mg of mannitol, 2974 mg of sterile water for injection, and Na ₂ HPO ₄ · 12H ₂ 0 26 mg were prepared and dispensed at 2 mL per ampoule.

Claims (8)

Pharmaceutical composition for the prevention or treatment of dementia, which contains the stalk extract as an active ingredient.
According to claim 1,
The stalk tree extract
Characterized in that the water, the lower alcohol having 1 to 6 carbon atoms, or a mixture of them, the stalk tree extract,
Pharmaceutical composition for preventing or treating dementia.
According to claim 1,
The stalk tree extract
Characterized in that the hedgehog extract with 30% to 95% ethanol or hot water extract,
Pharmaceutical composition for preventing or treating dementia.
According to claim 1,
The stalk tree extract
Characterized in that it is a henna extract or hot water extract with 70% ethanol,
Pharmaceutical composition for preventing or treating dementia.
According to claim 1,
The stalk tree extract
Characterized in that the fraction of the stalk extract by 30% to 95% ethanol is at least any one of acetate and butanol from the stalk extract by water, lower alcohol having 1 to 6 carbon atoms, or a mixed solvent thereof.
Pharmaceutical composition for preventing or treating dementia.
According to claim 1,
The myrtle extract is characterized in that it exhibits the inhibitory effect of beta amyloid aggregation or inflammation of the brain microglial cells,
Pharmaceutical composition for preventing or treating dementia.
According to claim 1,
The myrtle tree extract is characterized in that it exhibits the ability to inhibit vascular dementia,
Pharmaceutical composition for preventing or treating dementia.
According to claim 1,
The sash tree extract is characterized in that the extract of the leaves or stems of the sash tree,
Pharmaceutical composition for preventing or treating dementia.

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