KR20210031402A - Composition for Improving Dry Eye Syndrome Comprising Extract of Aralia - Google Patents

Abstract

The present invention relates to a composition for alleviating dry eye syndrome containing an extract of Aralia elata. The composition according to the present invention has almost no toxicity and side effects, thereby being able to be used to more safely prevent or alleviate dry eye syndrome. An objective of the present invention is to provide the composition for alleviating dry eye syndrome that can safely and effectively prevent or treat dry eye syndrome and can be formulated in various formulations.

Description

Composition for Improving Dry Eye Syndrome Comprising Extract of Aralia

The present invention relates to a composition for improving dry eye syndrome containing an Aralis tree extract, and more particularly, to a composition that can safely and effectively prevent or improve dry eye syndrome by containing an Aralis tree extract.

Dry eye syndrome is a symptom that shows symptoms such as glare, irritation of the eyes, drowsiness, foreign body sensation, burning sensation, and local itching. It is a symptom that occurs due to a decrease in the amount of tears in the eye or a change in components.

The most common cause of dry eye syndrome is aging, which is indicated by a decrease in the amount of secretion or a change in the state of tears due to normal aging. In addition, with diseases such as rheumatoid arthritis, Sjogren's syndrome, lupus, scleroderma, diabetes, and vitamin A deficiency, the secretion of tears decreases, resulting in dry eye symptoms. In addition, dry eye syndrome may be caused by chronic conjunctivitis, thyroid disease, menopausal female hormone reduction, and drug use.

Drugs that affect tear secretion include antibiotics, antihistamines, diuretics, antidiarrheal drugs, parasympathetic nerve blockers such as scoporamine, beta blockers for the treatment of hypertension, sleeping pills, contraceptives, some acne medications, some antidepressants, some anesthetics, etc. Temporary tear reduction may occur while taking.

In addition, as environmental factors, dry eye syndrome is likely to occur when the surrounding environment is dry, the eyes are irritated by smoke, dust, sunlight, wind, etc., or when the number of unconscious blinking while reading or using a computer decreases.

The most common countermeasure for dry eye syndrome is to use artificial tears to supply insufficient tear components, and there are various types such as solutions, ointments, and thin jelly forms. However, the method of using artificial tear fluid has only the effect of temporarily improving the symptoms of dry eye syndrome, and is insufficient to fundamentally alleviate the symptoms by inhibiting the progression of dry eye syndrome.

Korean Patent Registration No. 10-1507504 includes sulfasalazine or hydrophilic sulfasalazine; And hyaluronic acid, wherein the sulfasalazine or hydrophilized sulfasalazine and hyaluronic acid have a weight ratio of 0.01:1 to 50:1.An eye drop composition for the prevention or treatment of dry eye syndrome or related ophthalmic diseases or ophthalmic symptoms has been disclosed. However, the ophthalmic composition includes hyaluronic acid and has side effects such as congestion, eye irritation, and itchy eyelids.

In addition, there is a need to develop a treatment for dry eye syndrome that can be formulated into various formulations such as oral dosage forms in order to improve medication compliance.

Korean Patent Registration No. 10-1507504

It is an object of the present invention to provide a composition for improving dry eye syndrome that can safely and effectively prevent or treat dry eye syndrome and can be formulated in various ways.

One embodiment of the present invention relates to a pharmaceutical composition for the prevention or treatment of dry eye syndrome comprising an Aralis tree extract as an active ingredient.

Aralia elata is a plant belonging to the genus Aralia sp., and is a reed tree. Aralia are distributed in East Asia, mainly Manchuria and Sakhalin, and grow at the edge of forests nationwide in Korea. The bark, wood, and branches of Aralia have been dried and have been used as herbal medicinal materials since ancient times.

There are several types of triterpenoids, including saponins, in the bark of the Aralia tree, and several glycosides, including elatoside E, which have a hypoglycemic effect, elatoside F and oleanolic acid glycoside. It contains seeds, and also contains elatosides A and B, which inhibit ethanol absorption. In private and oriental medicine, the bark of Aralia is used for seawater, gastric cancer, diabetes and gastrointestinal disorders.

The Aralia tree extract can be prepared by performing an extraction process comprising extracting Aralia tree with an extraction solvent (primary extraction solvent) selected from the group consisting of water, an organic solvent, and a mixed solvent of water and an organic solvent. .

The organic solvent includes, but is not limited to, a _{C 1} -C _{4 alcohol, ethyl acetate, and the like.}

The C ₁ -C ₄ alcohol refers to a straight-chain or branched alcohol having 1 to 4 carbon atoms, and includes, for example, methanol, ethanol, n-propanol, isopropanol, n-butanol, and the like, but is not limited thereto. It is preferable to use ethanol as the alcohol.

The extraction process may be performed by extracting with a primary extraction solvent of about 1 to 20 times the weight of the aralia tree, preferably a mixed solvent of water and ethanol. About 70% ethanol aqueous solution may be used as the mixed solvent.

Extraction may be performed by a known extraction method, such as cold sedimentation, hot water extraction, ultrasonic extraction, reflux cooling extraction, or the like, but is not limited thereto. The extraction temperature may be adopted by a person skilled in the art in a variety of temperature ranges suitable for the extraction method, and may be performed at, for example, 0 to 120°C, but is not limited thereto. The extraction time is different depending on the extraction method, and a person skilled in the art can adopt an appropriate extraction time. For example, the extraction time may vary depending on the temperature, but is not limited thereto, but may be performed once or multiple times in the range of about 1 hour to 10 days. The extraction time can be reduced to about 1 to 3 hours when pressure is applied. The pressure may be usually 1.2 to 1.5 kg/cm 2, at which time the temperature reaches 110 to 120°C. Preferably, the extraction may be performed by extraction with the primary extraction solvent described above for about 48 to 71 hours at room temperature.

The extract obtained by performing the extraction with the primary extraction solvent is obtained in a liquid form from which impurities are removed by filtration or centrifugation according to a conventional method, or the obtained liquid extract is concentrated under reduced pressure and/or dried according to a conventional method. It can be obtained in powder form.

In addition, the extraction process may further include a step of obtaining a fraction having a high content of an active ingredient, if necessary. That is, after dispersing the extract obtained by extraction with the primary extraction solvent in water, extraction with one or more extraction solvents selected from the group consisting of an appropriate secondary extraction solvent, such as chloroform, ethyl acetate and butanol, to obtain a fraction, The content of active ingredients in the resulting extract can be further increased. In particular, the content of the active ingredient in the butanol extract, that is, the butanol fraction, is the highest among the secondary extraction solvents.

One embodiment of the present invention relates to a pharmaceutical composition for the prevention or treatment of dry eye syndrome comprising as an active ingredient chlorogenic acid isolated from the extract of Aralia elata.

The chlorogenic acid can be obtained by column chromatography on the extract of Aralia.

The extract of the Araliaceae tree of the present invention and the chlorogenic acid isolated therefrom were found to be excellent in inhibiting cell damage in the hyperosmotic stress model and the oxidative stress model as a dry eye syndrome cell model, and it was confirmed that it can prevent or ameliorate dry eye syndrome (Experimental Examples 2 and 4).

In the present invention, TonEBP, also known as NFAT5, which is a neurotransmitter, and NF-kB (p65) were _{increased in a stressed dry eye cell model by treatment with sodium chloride (NaCl) or H 2} O ₂ , and this was increased in a cell model of Araliaceae extract and chlorogenic acid ( It was confirmed that CGA) can be reduced (Experimental Examples 3 and 5).

In addition, it was confirmed that the extract of Aralis trees of the present invention exhibits an effect of improving dry eye syndrome within 1 month without side effects when applied to the human body (Experimental Example 6).

Accordingly, the extract of Aralia serrata of the present invention and the chlorogenic acid isolated therefrom can be effectively used in a pharmaceutical composition for preventing or treating dry eye syndrome.

The pharmaceutical composition according to the present invention may contain, in addition to the extract of Aralia, another treatment for dry eye syndrome.

The pharmaceutical composition according to the present invention can be administered orally (e.g., ingestion or inhalation) or parenterally (e.g., injection, transdermal absorption, rectal administration, eye drop), and the injection may be, It may be intravenous, subcutaneous, intramuscular, or intraperitoneal injection. According to the route of administration, the pharmaceutical composition according to the present invention includes powders, granules, tablets, capsules, fine subtilae, powders, sublingual tablets, suppositories, ointments, emulsions, solutions, suspensions, emulsions, It can be formulated as a syrup, ophthalmic solution (eye drops), injection, spray, and the like. The various types of pharmaceutical compositions according to the present invention may be prepared by known techniques using a pharmaceutically acceptable carrier commonly used in each formulation. Examples of pharmaceutically acceptable carriers include excipients, binders, disintegrating agents, lubricants, preservatives, antioxidants, isotonic agents, buffers, coating agents, sweetening agents, solubilizing agents, bases, dispersing agents, wetting agents. , Suspending agents, stabilizers, coloring agents, and the like.

The pharmaceutical composition according to the present invention may vary depending on the form of the drug, but may contain about 0.01 to 95% by weight of the Aralia extract or chlorogenic acid.

The specific dosage of the pharmaceutical composition of the present invention may vary depending on the type, weight, sex, degree of disease, judgment of a doctor, and the like of the mammal including the person to be treated. Preferably, 10 to 200 mg of active ingredient per 1 kg of body weight per day for oral administration, and 0.01 to 10 mg of active ingredient per 1 kg body weight per day for parenteral administration. The total daily dosage may be administered at once or divided into several times depending on the degree of disease, judgment of a doctor, and the like.

One embodiment of the present invention relates to a health functional food for preventing or improving dry eye syndrome comprising an Aralis tree extract as an active ingredient.

One embodiment of the present invention relates to a health functional food for preventing or improving dry eye syndrome comprising chlorogenic acid isolated from the Aralis tree extract as an active ingredient.

There is no particular limitation on the type of health functional food according to the present invention, and is in the form of oral preparations such as powders, granules, tablets, capsules, solutions, suspensions, emulsions and syrups, or candy, confectionery, gum, ice cream, and noodles. It can be added to general foods such as bread, beverages, etc.

The health functional food of the present invention can be prepared by appropriately using a filler, an extender, a binder, a wetting agent, a disintegrant, a sweetener, a fragrance, a preservative, a surfactant, a lubricant, an excipient, and the like in a conventional manner according to the form.

In the manufacture of the health functional food, the content of Aralis tree extract or chlorogenic acid varies depending on the form of the health functional food, but is about 0.01 to 95% by weight.

Since the health functional food of the present invention has almost no toxicity and side effects, it can be safely used even when taken for a long time for prophylactic purposes.

Aralia extract according to the present invention and the chlorogenic acid isolated therefrom are shown to have an excellent effect of inhibiting cell damage in a dry eye cell model, and can exhibit an effect of improving dry eye syndrome without side effects when applied to the human body. It can be effectively used in medical compositions or health functional foods.

In addition, the Araliaceae extract according to the present invention has almost no toxicity and side effects, so it can be used to more safely prevent or improve dry eye syndrome.

1 is a graph showing the results of investigating the effect of various levels of hyperosmotic stress on cell survival.
2 is a graph showing the results of investigating the effect of the Aralis tree extract on the cell survival rate by treating the Aralis tree extract by concentration in the hyperosmotic stress cell model.
Figure 3 shows the results of investigating the effect on the cell survival rate when pretreatment of Aralia extract (AE) and chlorogenic acid (CGA) analyzed as a major component of Aralia extract to cells by concentration, and a high osmotic stress of 500 mOsm. This is the graph shown.
Figure 4 is a graph showing the results of investigating the effect on the cell survival rate when cells were pretreated with 5 µg of Aralia serrata extract (AE) and 300 µM of chlorogenic acid (CGA), and differently given high osmotic stress at 450 mOsm and 500 mOsm, respectively. to be.
Figure 5 is a result of analyzing the expression of TonEBP and p-NF-kB (p65) by Western blotting when pre-treatment of Aralia extract (AE) and chlorogenic acid (CGA), respectively, and applying high osmotic stress.
6 is a graph showing the results of examining the effect on cell viability by treating _{H 2} O ₂ by concentration in order to select an appropriate concentration of _{H 2} O ₂ in the oxidative stress model.
7 is a graph showing the results of investigating the change in cell viability in the group and the group pretreated with Aralia extract (AE) and chlorogenic acid (CGA) simultaneously with oxidative stress.
Figure 8 is a result of analyzing the expression of TonEBP and p-NF-kB (p65) by Western blotting when oxidative stress was applied with _{H 2} O ₂ after each pretreatment of Araliaceae extract (AE) and chlorogenic acid (CGA) to be.
9 is a graph showing the results of monitoring the efficacy and side effects for those who have taken Aralis tree extract for more than 1 month.

Hereinafter, the present invention will be described in more detail by examples. These examples are for illustrative purposes only, and it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

Example 1: Preparation of Aralia hot water extract

Aralia 2018 was purchased and the branches were crushed to take 200 g, and 1,600 ml of distilled water was added thereto, followed by hot water extraction at 100°C for 8 hours, filtration and concentration, and drying to obtain a hot water extract.

Example 2: Preparation of ethanol extract and chlorogenic acid of Aralia elata

Aralia 2018 was purchased, the branches were crushed, 200 g was taken, and 1.2 L of 70% ethanol was added, and the mixture was eluted for 48 to 72 hours while stirring occasionally at room temperature. The eluate was decanted, and 1.2ℓ of 70% ethanol was added to the remaining residue again, and the same method was repeated to elute again.

The re-elutant was combined with the first eluate according to the gradient method, ethanol was recovered in a reflux cooling device, the remaining eluate was centrifuged to remove the precipitate, and the remaining supernatant was filtered. Hexane and chloroform were added to the filtrate to remove resin, protein, and fiber, and the remaining liquid was lyophilized to powder.

Using the 70% ethanol extract of Aralia elata obtained above, it was fractionated into four fractions, that is, a chloroform fraction, an ethyl acetate fraction, a butanol fraction, and a water fraction by a solvent extraction method. Specifically, after dissolving the 70% ethanol extract of Aralia elata in water, chlorophyll and other impurities are removed using petroleum ether, and the remaining liquid is sequentially chloroform (CHCl ₃ ), ethyl acetate, butanol ( BuOH) and a water layer were divided into powder, respectively.

The butanol fraction was subjected to high-performance liquid chromatography (HPLC). At this time, μbondapak C ₁₈ (3.9×300mm) was used as the column, and 2% acetic acid:methanol was used as an elution solvent in a gradient mode (10: 0 → 2: 8). A small fraction was eluted by performing HPLC for 30 minutes at a temperature of 40° C. at a flow rate of 1 ml/min. As a result of identification of the obtained small fraction using LC-MS, it was confirmed that the small fraction with the highest content was chlorogenic acid.

Experimental Example 1: Analysis of the components of the extract of the present invention Aralia

Using the test substance standard solution, the LC-MS instrument conditions without interfering substances in the retention time of the test substance were established as shown in Table 1 below.

According to the analysis conditions of Table 1, the results of the component analysis of the Araliaceae extract prepared in Example 2 are shown in Table 2 below.

Test substance Index component content(%) Content (g/kg) Aralia extract Caffeic acid 0.0322 0.322 Chlorogenic acid 0.464 4.64 3,4-dihydroxybenzoic acid 0.394 3.94

From Table 2, it was found that the content of phenolic substances in the extract of Aralia tree prepared in Example 2 was high. In particular, it was confirmed that the highest level of chlorogenic acid (CGA) was contained in the Aralinium tree extract.

Experimental Example 2: High osmotic stress model experiment

The efficacy test for dry eye syndrome was performed on HCEC (human corneal epithelial cell) as a cell model. As a dry eye syndrome model, a hyperosmolar stress model was used.

The hyperosmotic stress model was induced by treating cells with sodium chloride (NaCl).

(1) Determination of high osmotic stress level

Hyperosmotic stress induces the death of epithelial cells in the cornea or lens, causing cell damage. In this experiment, in order to determine the level of hyperosmotic stress to be used in the cell experimental model, the effects of various levels of hyperosmotic stress on the cell viability were investigated.

In order to confirm the cell viability, MTT assay was used.

MTT analysis was performed as follows.

The cell suspension was counted using a hemacytometer, and then ^{500 μl at a concentration of 1×10 5} cells/ml was put into each well of a 24-well plate and incubated at 37° C. for 24 hours. Thereafter, the concentration of NaCl was adjusted to induce various levels of high osmotic stress, and the samples were treated by concentration and further incubated for 24 hours. After removing the sample, 200 µl of MTT solution was added to each well, and light was blocked. After incubation for 4 hours, the MTT dilution was carefully removed. 200 µl of DMSO was added to each well and shaken with a plate shaker for 15 to 20 minutes while blocking the light. Absorbance was measured at 570 nm using an ELISA reader. 100 μl of 96 wells were divided into two wells, and absorbance was measured at 570 nm using an ELISA reader. This absorbance represents the amount of MTT reduced by the cells and is proportional to the number of viable cells present in each well.

The results are shown in FIG. 1.

1, it can be seen that when the high osmotic stress is 500 mOsm, the cell viability decreases, and when the high osmotic stress is 550 mOsm, the cell viability decreases to a level of 50%.

Therefore, the high osmotic stress level for the experiment was determined to be 500 mOsm.

(2) Determination of the concentration of Aralis tree extract

A high osmotic stress was applied to the cells at 500 mOsm, and at this time, the dry powdery Aralis tree extract prepared in Example 1 was treated with each concentration to investigate the effect of the Aralis tree extract on cell viability.

In order to confirm the cell viability, MTT analysis was used.

MTT analysis was performed in the same manner as in the determination of the high osmotic stress level.

The results are shown in FIG. 2.

Through FIG. 2, it was confirmed that the best efficacy appeared when the extract of Aralia elata was treated with 5 μg.

(3) Efficacy evaluation by concentration of Aralis tree extract (AE) and chlorogenic acid (CGA)

Cells were pretreated with dry powdery Aralis tree extract (AE) prepared in Example 1 and chlorogenic acid (CGA) analyzed as a main component of Aralis tree extract prepared in Example 2 by concentration, and had high osmotic pressure. When stress was applied at 500 mOsm, the effect of Araliaceae extract and CGA on cell viability was investigated.

In order to confirm the cell viability, MTT analysis was used.

MTT analysis was performed in the same manner as in the determination of the high osmotic stress level.

The results are shown in FIG. 3.

3, it can be seen that the cell damage inhibitory effect is the most excellent in 5 µg of Aralis tree extract (AE) and 300 µM of CGA.

(4) Efficacy evaluation of Aralia tree extract (AE) and chlorogenic acid (CGA) according to high osmotic stress conditions

When cells were pretreated with 5 μg of dried powdery Aralis tree extract (AE) prepared in Example 1 and 300 μM of CGA, and the high osmotic stress was different at 450 mOsm and 500 mOsm, respectively, the cell viability of Aralis tree extract and CGA The effect on the was investigated.

In order to confirm the cell viability, MTT analysis was used.

MTT analysis was performed in the same manner as in the determination of the high osmotic stress level.

The results are shown in FIG. 4.

Through FIG. 4, both the inhibitory effect of cell damage inhibitory effect of Aralia tree extract (AE) and CGA under two hyperosmotic stress conditions was confirmed.

Experimental Example 3: Mechanism of action experiment in a high osmotic stress model

TonEBP (Tonicity response element binding protein) is known to be the most important transcription factor widely expressed in the mechanism by which cells protect themselves from osmotic stress. TonEBP is regulated by tonicity outside the cell, and under hypertonic conditions, TonEBP level and activity rise in the cell nucleus. In addition, TonEBP is involved in overexpression and activation of selected signaling molecules and enzymes including aldose reductase (AR) and nuclear factor-kappa B (NF-kB).

In this experimental example, in order to confirm the mechanism of action in the high osmotic stress model, Aralia extract (AE) 5㎍ (A5) and 10㎍ (A10), CGA 100 μM (C100) and 300 μM (C300) were pretreated for 24 hours, respectively, and TonEBP and p- The expression of NF-kB (p65) was investigated. Proteins were separated by SDS-PAGE, and the separated proteins were analyzed by western blotting.

Protein separation through SDS-PAGE was performed as follows.

The cell suspension was counted using a hemacytometer, and then 2 ml in each well of a 6-well plate at a concentration of ^{1×10 5 cells/ml, and incubated at 37° C. for 24 hours.} Thereafter, the samples were treated by concentration and incubated for a desired time. The sample was removed and washed with PBS. Cells were scraped with a cell scraper and collected in 5 ml falcon tubes. Centrifugation was performed for 3 minutes at 3,000 rpm in a desk top centrifuge. After removing the PBS, 20 µl of a lysis buffer was added per tube, and then the cells were lysed by pipetting. The tube was placed on ice for 20 minutes. Then, the sample was sonicated using a sonicator. After ultrasonic grinding for 10 to 30 seconds, the process of placing on ice for 1 minute was repeated 3 times. After this, it was put on ice for 20 minutes. After centrifugation at 4° C. for 30 minutes at a rate of 12,000 g, the supernatant was transferred to a new tube and quantified at 1:15.

Western blotting was performed as follows.

SDS-PAGE samples were prepared using a 4-fold sample buffer by calculating the values quantified in the SDS-PAGE. A gel was prepared according to the size of the antibody. The sample was boiled for 3 minutes, cooled in ice water for 3 minutes, and then centrifuged. The sample was loaded on the gel and proceeded to 6 mA (per plate) in the stacking gel, and increased to 9 to 10 mA in the separating gel. When the desired size was reached, it was stopped and the proteins in the gel were transferred to the NC membrane using a transfer. Then it was blocked with 3% BSA and the desired Ab was incubated overnight at 4°C. The next day, after incubation with the secondary Ab, the band was confirmed using an ECL solution.

The results are shown in FIG. 5.

5, it was confirmed that TonEBP increased protein expression when hyperosmotic stress was applied, and then decreased expression in both groups pretreated with Aralia extract (AE) and CGA of the present invention, p-NF-kB ( p65) was confirmed to show efficacy by decreasing its expression in the group pretreated with 5 µg of Aralis tree extract for 1 hour and the group pre-treated with 100 µM of CGA.

Experimental Example 4: Oxidative Stress Model Experiment

In this experiment, an oxidative stress model was used as another dry eye syndrome model, and the efficacy of the extract of Aralinium tree to treat dry eye syndrome was confirmed.

Efficacy experiments on dry eye syndrome were performed on HLE-B3 (human lens epithelial cells) as a cell model.

The oxidative stress model was induced by treating _{cells with H 2} O _2.

(1) H ₂ O ₂ Calculation of the appropriate concentration of

Selecting a suitable concentration of H ₂ O ₂ the survival rate was confirmed by treating the cells with H ₂ O ₂ at different concentrations in order. In order to confirm the cell viability, MTT analysis was used.

MTT analysis was performed in the same manner as in the determination of the high osmotic stress level.

The results are shown in FIG. 6.

6, since about 50% cell viability was confirmed at 0.1 mM, the concentration used in the experiment was set to 0.1 mM.

(2) Effect of Aralia serrata extract and chlorogenic acid (CGA) in oxidative stress model

The change in cell viability was investigated in the group treated with the dried powdery Aralis tree extract (AE) and CGA prepared in Example 1 at the same time with oxidative stress and the pretreated group. In order to confirm the cell viability, MTT analysis was used.

MTT analysis was performed in the same manner as in the determination of the high osmotic stress level.

The results are shown in FIG. 7.

Through FIG. 7, it was confirmed that the group pre-treated with Aralia extract (AE) and CGA had higher effect than the group treated at the same time.

Experimental Example 5: Mechanism of action experiment in oxidative stress model

Oxidative stress To confirm the mechanism of action in the model, When oxidative stress was applied to HLE-B3 (human lens epithelial cell) wild type (WT) and NFAT5 (=TonEBP) knockout (KO) cell lines with H ₂ O ₂ , the Aralia extract of the present invention ( AE) After treatment with 5㎍ (A5) and 100 μM (C100) and 300 μM (C300) of chlorogenic acid (CGA), which are major components of Araliaceae extract, respectively, the expression of TonEBP and p-NF-kB (p65) was investigated. I did. Proteins were separated by SDS-PAGE, and the separated proteins were analyzed by western blotting.

SDS-PAGE and Western blotting were performed in the same manner as in Experimental Example 3.

The results are shown in FIG. 8.

8, in the wild type (Wild Type, WT), TonEBP and p-NF-kB tend to decrease in the group treated with chlorogenic acid (CGA), which is a major component of the present invention, Aralia extract (AE) and Aralia extract. It can be seen that it represents.

In addition, through FIG. 8, TonEBP and p-NF-kB did not appear in the KO group because TonEBP was knocked out, but TonEBP and p-NF-kB increased when stress was applied even though TonEBP and p-NF-kB were not present. When treated with Aralia extract (AE) of the present invention and chlorogenic acid (CGA), which is a major component of Aralia extract, Appeared to decrease.

From the above results, it was shown that p-NF-kB was not affected as a sub-step of TonEBP, and the treatment with chlorogenic acid (CGA), which is a major component of the Aralis tree extract (AE) and Aralis tree extract of the present invention, was performed with TonEBP and p- It was found that each influenced the expression of NF-kB.

Experimental Example 6: Evaluation of the efficacy of the extract of the present invention on improving dry eye in human body

Thirty-six volunteers were given the extract of the present invention for 1 to 3 months to monitor the efficacy or side effects.

The number of subjects was 36, which was taken by 18 men and 18 women. Distribution by age was shown in Table 3 below.

age personnel 10 units (10-19) 2 20s (20-19) 3 30s (30-39) 2 40s (40-49) 6 50s (50-59) 8 60s (60-69) 10 70s (70-79) 3 80 cars (80-19) 2 Sum 36

In order to confirm the safety of the Aralis tree extract, a 28-day repeat toxicity test was performed in rats for the Aralis tree extract of Example 1. As a result, it was confirmed that the NOAEL value is more than 2,000mg/kg, which is a very safe material.

Based on the results of the safety test, the Aralia extract was set at 200 mg/poro, the single dose was 200 mg × 2 packets = 400 mg, and the daily dose was 400 mg × 2 times = 800 mg (based on 60 kg of adults).

Responses to questions about efficacy and side effects were monitored in subjects who took Aralinium extract for more than 1 month. The monitoring items were age, sex, duration of taking Aralinium extract, and efficacy or side effects during or after taking it.

The results are shown in Table 4 and FIG. 9 below.

number name gender age Period of use Perceived efficacy after use Reduced eye strain Vision
Improving Improvement of dry eye Improvement of cataracts, etc. One Lee OO male 42 3 months ○ 2 Park OO female 63 3 months ○ ○ ○ 3 Kim OO female 59 3 months ○ ○ 4 Jang OO male 61 3 months ○ 5 Shin OO male 63 3 months ○ ○ 6 Kim OO female 66 3 months ○ ○ 7 Kwon OO female 57 3 months ○ ○ ○ 8 Kim OO male 59 3 months ○ ○ 9 Yoo OO female 80 3 months ○ ○ 10 Jung OO female 36 3 months ○ ○ 11 Shin OO female 40 3 months ○ 12 Kang OO male 47 3 months ○ ○ 13 Lee OO female 33 3 months ○ 14 Jo OO male 54 3 months ○ ○ 15 Park OO female 53 3 months ○ ○ 16 Shin OO female 60 3 months ○ ○ 17 Kim OO male 62 3 months ○ 18 Jung OO male 48 3 months ○ ○ 19 Lee OO male 45 3 months ○ 20 Lim OO male 64 3 months ○ 21 Hong OO female 55 3 months ○ 22 Park OO male 62 3 months ○ 23 Lee OO female 18 3 months ○ ○ 24 Lim OO female 25 3 months ○ ○ ○ 25 Baek OO male 52 3 months ○ ○ 26 Uhm OO female 48 3 months ○ ○ 27 Lee OO female 78 3 months ○ ○ ○ 28 Kim OO male 61 3 months ○ 29 Kim OO male 80 3 months ○ ○ ○ 30 Lee OO female 19 3 months ○ ○ 31 Kang OO female 20 3 months ○ 32 Lee OO male 24 3 months ○ 33 Sung OO male 57 3 months ○ ○ 34 Lee OO male 75 3 months ○ ○ 35 Oh OO male 73 2 months ○ 36 Jo OO female 61 2 months ○ ○

From Table 4 and FIG. 9, it can be seen that the overall efficacy when taking the Aralinium tree extract shows improvement of vision, improvement of dry eye, reduction of eye fatigue, improvement of cataract symptoms, and the like.

When taking Aralia extract twice a day after morning and evening meals, the time taken to show efficacy varies from person to person, but ranged from 1 week to 1 month. In addition, no side effects were observed due to the intake of Aralis tree extract.

Formulation example:

The following formulation was prepared using the Aralia tree extract prepared in Example 1. However, the following formulation examples are only illustrative of the present invention, and the content of the present invention is not limited thereby.

Formulation Example 1: Preparation of tablets

250 mg of dried powdered Aralia serrata extract prepared in Example 1 was used as an excipient for direct injection of 260 mg of lactose and Avicel (microcrystalline cellulose) 35 mg, a disintegration aid of sodium starch glyconate 15 mg, and a binder for direct injection of L-HPC (Low -hydroxypropylcellulose) 80 mg and put in a U-type mixer and mixed for 20 minutes. After the mixing was completed, an additional 10 mg of magnesium stearate, a lubricant, was added and mixed for 3 minutes. Tablets containing 250 mg of extract per tablet were prepared by tableting and film-coating after a quantitative test and a humidity test.

Formulation Example 2: Preparation of syrup

An appropriate amount of sucrose was dissolved in a certain amount of water, 80 mg of paraoxymethylbenzoate and 16 mg of paraoxypropylbenzoate were added thereto as preservatives, and 4.5 g of dried powdery Araliaceae extract prepared in Example 1 was added thereto, After completely dissolving while maintaining at 60° C., it was cooled and distilled water was added to make 150 ml to prepare a syrup.

Formulation Example 3: Preparation of capsules

A capsule formulation was prepared by mixing 300 mg of the dried powdered Aralis tree extract prepared in Example 1 with 200 mg of lactose as a carrier and filling it into a hard gelatin capsule.

Formulation Example 4: Preparation of beverage

After dissolving 500 mg of the dried powdered Araliaceae extract prepared in Example 1 in an appropriate amount of water, vitamin C as an auxiliary component, citric acid, sodium citrate, and high fructose as a mating agent were added in an appropriate amount, and an appropriate amount of sodium benzoate as a preservative. After adding, water was added to make the total amount to 100 ml to prepare a beverage.

Formulation Example 5: Preparation of injection

200 mg of the dried powdered Araliaceae extract prepared in Example 1 was heated and dissolved in 200 mg of physiological saline containing 1% polyoxyethylene hydrogenated castro oil to prepare an injection containing the extract at a concentration of 0.1% by weight. .

Formulation Example 6: Preparation of eye drops

An eye drop was prepared by heating and dissolving 200 mg of the dried powdery Araliaceae extract prepared in Example 1 in physiological saline.

Claims (12)

A pharmaceutical composition for the prevention or treatment of dry eye syndrome comprising an Aralis tree extract as an active ingredient. The pharmaceutical according to claim 1, wherein the extract of Aralia is produced by performing an extraction process comprising extracting Aralis tree with a primary extraction solvent selected from the group consisting of water and a mixed solvent of water and an organic solvent. Composition. The pharmaceutical composition according to claim 2, wherein the organic solvent is ethanol. The method of claim 2, wherein in the extraction step, the extract obtained by extraction with the primary extraction solvent is dispersed in water, and then extracted with one or more secondary extraction solvents selected from the group consisting of chloroform, ethyl acetate, and butanol to obtain a fraction. A pharmaceutical composition further comprising the step of obtaining. The pharmaceutical composition according to claim 1, having a dosage form selected from the group consisting of powders, granules, tablets, capsules, solutions, suspensions, emulsions, syrups, ophthalmic preparations and injections. A pharmaceutical composition for preventing or treating dry eye syndrome comprising chlorogenic acid as an active ingredient. Health functional food for the prevention or improvement of dry eye syndrome containing Aralis tree extract as an active ingredient. The health of claim 7, wherein the Aralis tree extract is prepared by performing an extraction process comprising extracting Aralis tree with a primary extraction solvent selected from the group consisting of water and a mixed solvent of water and an organic solvent. Functional food. The health functional food according to claim 8, wherein the organic solvent is ethanol. The method of claim 8, wherein in the extraction step, the extract obtained by extraction with the primary extraction solvent is dispersed in water, and then extracted with one or more secondary extraction solvents selected from the group consisting of chloroform, ethyl acetate, and butanol to obtain a fraction. Health functional food further comprising the step of obtaining. The health functional food according to claim 7, having a formulation selected from the group consisting of beverages, powders, granules, tablets, capsules, solutions, suspensions, emulsions and syrups. Health functional food for preventing or improving dry eye syndrome containing chlorogenic acid as an active ingredient.

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