WO2018190638A1

WO2018190638A1 - Composition for preventing or treating corneal diseases, containing glycine max extract

Info

Publication number: WO2018190638A1
Application number: PCT/KR2018/004253
Authority: WO
Inventors: 김경아; 강태겸; 강석우; 오상록; 정상훈
Original assignee: 한국과학기술연구원
Priority date: 2017-04-12
Filing date: 2018-04-11
Publication date: 2018-10-18
Also published as: KR101838142B1

Abstract

Disclosed in the present specification are: a composition for preventing or treating corneal disease, containing a Glycine max extract as an active ingredient; and a dietary supplement composition for alleviating corneal injury, containing a Glycine max extract as an active ingredient. The Glycine max extract has effects of inhibiting corneal cell damage, protecting corneal cells, and preventing, alleviating, or treating dry eye syndrome and corneal abrasion.

Description

Composition for the prevention or treatment of corneal disease comprising rat eye extract

The present disclosure discloses a pharmaceutical composition for preventing or treating a corneal disease comprising a rat eye extract as an active ingredient and a health food composition for improving corneal damage comprising a mouse head extract as an active ingredient.

The cornea is a transparent, bloodless tissue on the anterior surface of the eye, commonly called the black jerky. The cornea is a transparent tissue that not only protects the eyes from the outside but also allows the light to pass through and bend, and plays a major role in the refraction and transmission of light. The cornea is composed of five layers of corneal epithelium, Bowman's membrane, corneal stroma, posterior border plate (desme membrane) and corneal endothelium.

The corneal surface is directly exposed to the outside, making it vulnerable to trauma and scratches. In particular, corneal damage may occur frequently in patients suffering from diseases such as dry eye syndrome caused by a significant decrease in tear protection due to the lack of tears. If the cornea is damaged, irritation symptoms such as irritation, foreign body or dryness, as well as if the damage is severe may develop keratitis. Thus, to keep the eye healthy and preserve vision, it is essential to prevent corneal damage, to prevent minor corneal damage from worsening, or to treat damaged corneas.

Dry eye is a condition in which tears evaporate quickly due to lack of tears or an imbalance of tear components, causing damage to the tear film and cornea surface. It was defined as a disease accompanied by inflammatory changes in the tissues involved in the tear secretion process and osmotic pressure of tears. Damage to the tear film and corneal surface causes the eye surface to dry out, causing eye discomfort and irritation. Dry eye syndrome may be caused by a decrease in androgen, which controls tear secretion, living in a dry environment with high room temperature, and wearing contact lenses in the presence of systemic diseases such as Sjogren's syndrome, and the use of computers, smartphones and tablets. Etc. In addition, dry eye syndrome continues to increase not only for the elderly but also for the young people due to the increase in the elderly population, the performance improvement of computer-related devices, the development of communication technology, and the growth of the industry.

Currently, the main treatment of dry eye depends on various artificial tears. Recently, there is a report that the inflammatory response is also associated with the occurrence of dry eye syndrome, immunosuppressants such as cyclosporin (cyclosporin) has been developed as an eye drop and used in the treatment, but the exact pathophysiology is not known yet. Related prior arts are Korean Patent Publication No. 10-1662720.

In one aspect, an object of the present disclosure is to provide a pharmaceutical composition for preventing or treating corneal damage and / or corneal disease, which comprises a rat bean extract as an active ingredient.

In another aspect, the present disclosure aims to provide a health food composition for preventing or improving corneal damage and / or corneal disease, including the rat bean extract as an active ingredient.

In one aspect, the technology disclosed herein provides a pharmaceutical composition for the prevention or treatment of corneal disease comprising a rat eye extract as an active ingredient.

In an exemplary embodiment, the rat eye extract may be extracted with one or more solvents selected from the group consisting of water and C ₁ to C ₄ lower alcohols.

In an exemplary embodiment, the rat bean extract may be an ethanol extract.

In an exemplary embodiment, the composition may be to prevent or treat dry eye syndrome or corneal abrasions.

In an exemplary embodiment, the composition may be one having a corneal epithelial cell death inhibition and corneal epithelial layer protective effect.

In another aspect, the technology disclosed herein provides a health food composition for corneal damage improvement comprising a rat eye extract as an active ingredient.

In an exemplary embodiment, the rat bean extract may be an ethanol extract.

In an exemplary embodiment, the composition may be to improve dry eye or corneal abrasions.

In one aspect, the technology disclosed herein has the effect of providing a pharmaceutical composition for preventing or treating corneal damage and / or corneal disease, including rat bean extract as an active ingredient.

In another aspect, the technology disclosed herein has the effect of providing a health food composition for preventing or ameliorating corneal damage and / or corneal disease, including rat bean extract as an active ingredient.

In another aspect, the rat eye extract disclosed herein has the effect of inhibiting corneal cell damage, protecting corneal cells, preventing, ameliorating or treating dry eye, corneal abrasions.

1A and 1B show corneal fluorescein staining results in a BAC-induced corneal injury animal model according to an experimental example of the present specification ( *** p <0.001 ).

Figure 2 shows the tear film breakdown time measurement results in the corneal injury animal model induced by BAC according to an experimental example of the present specification ( *** p <0.001 ).

Figure 3 shows the results of hematoxylin eosin staining the cross-section of the cornea to observe the corneal surface and corneal epithelial cells in BAC-induced corneal injury animal model according to an experimental example of the present specification.

4A and 4B show the results of TUNEL assays for confirming corneal epithelial cell death in BAC-induced corneal injury animal models according to one experimental example of the present specification ( *** p <0.001 ).

Hereinafter, the present invention will be described in detail.

In one aspect, the technology disclosed herein provides a composition for preventing, ameliorating and / or treating a corneal disease or corneal injury comprising a rat eye extract as an active ingredient.

In another aspect, the techniques disclosed herein include a method for preventing corneal disease or corneal injury comprising administering to the subject in need thereof an amount of rattan extract effective in preventing, ameliorating and / or treating corneal disease or corneal injury, Provided are methods for improvement and / or treatment.

In another aspect, the technology disclosed herein provides the use of a rattan extract for preparing a composition for preventing, ameliorating and / or treating a corneal disease or corneal injury.

In another aspect, the techniques disclosed herein provide a rat eye extract for use in the prevention, amelioration and / or treatment of corneal disease or corneal injury.

In another aspect, the techniques disclosed herein provide a non-therapeutic use of a soybean extract for use in the prevention, amelioration and / or treatment of corneal disease or corneal injury.

In one exemplary embodiment, the composition may be a pharmaceutical composition or health food composition.

In one exemplary embodiment, the rat eye extract may be applied or administered to the subject in the form of a pharmaceutical composition or health food composition.

In an exemplary embodiment, the rat eye extract may be to be applied or administered to the eye of the subject.

The compositions disclosed herein have the effect of preventing corneal damage, preventing minor corneal damage from worsening, or ameliorating or treating a damaged cornea.

As used herein, the term "active ingredient" alone refers to a component that may exhibit the desired activity alone or together with a carrier having no activity.

As used herein, "corneal disease" refers to a disease occurring in the cornea or a disease caused by corneal injury, such as dry eye, corneal abrasion.

As used herein, “corneal injury” refers to damage to the corneal tissue of the eye, including but not limited to, pathogens, inflammation, physical stimuli (eg, contact lenses, ultraviolet light), chemical stimuli (eg, drugs), nerve damage Or it means that damage to the cornea by the accumulation of fatigue, etc., may be accompanied by symptoms such as pain, hyperemia, corneal haze, glare, foreign body, dry feeling. In addition, the corneal damage may be damage caused to any one of corneal epithelium, Bowman's cornea, corneal parenchyma, posterior border plate and corneal endothelial site, specifically, the rat eye extract has an excellent protective effect against corneal epithelial damage.

Dry eye syndrome is an eye disorder in which the eye's surface is damaged, the eye gets cold and there are irritating symptoms such as irritation, foreign body, and dryness due to lack of tears, excessive evaporation of tears, or poor balance of tear components. Say Rat eye extract according to the present disclosure has the effect of increasing the stability of the tear film and inhibiting corneal damage and protecting the surface of the cornea to prevent, improve or treat dry eye syndrome.

Corneal abrasion is a phenomenon in which the corneal epithelial layer of the eye is lost, for example, the epithelial layer is scratched or peeled off. The rat eye extract according to the present invention has an effect of protecting corneal surface and corneal epithelial layer by reducing corneal scratch and inhibiting corneal epithelial cell degeneration or corneal epithelial cell death.

Rhynchosia Nulubilis ) is a perennial perennial plant of legumes. It is a plant native to Sanya all over Korea. It is also called Seomoktae and Yeodu, and its stem and leaves are brown. In July, yellow flowers bloom and grow in oval pods with black and round fruits of 5-7 mm diameter.

In the old document `` Primary herb '', rat eye is "warm in nature, sweet in taste, and no poison. It is better to use light and soft, small soybeans (dungdue) as a medicine. Inhibits, activates blood circulation and detoxify ". Rats have long been used to prevent neuralgia, kidney disease and senile dementia. Rats have not only higher isoflavones than yellow soybeans, but also cyanidin-3-glucosides among glycidin and anthocyanin, which have excellent antioxidant effects on the seedlings. It is known to be effective in the prevention and treatment of cerebrovascular and heart diseases.

In the present specification, the rat eye extract may be a powder of the rat eye itself or a powder thereof, and includes all forms by processing such as the crude extract of the rat eye bean, for example, drying, concentration, fractionation, and fermentation. I mean.

In one exemplary embodiment, the rat eye extract may be prepared by the following steps, but is not limited thereto.

1) extracting by adding an extraction solvent to the rat eye;

2) filtering the extract of 'step 1)'; And

3) preparing an extract by concentrating the filtered extract of 'step 2)' under reduced pressure.

The rat eye extract 4) may further be prepared by further extracting the extract of 'step 3)' with an organic solvent to prepare a fraction.

The mouse eye bean of step 1) can be used without limitations such as cultivated, harvested or commercially available.

As the extraction solvent of step 1), it is preferable to use a solvent selected from water, an alcohol or a mixture thereof, preferably C ₁ to C ₆ , or C ₁ to C ₄ lower alcohol or a mixed solvent thereof. Or more preferably extracted with an aqueous solution of ethanol, but is not limited thereto. In addition, the amount of the extraction solvent is preferably 1 to 20 times the weight of rat eye, more preferably 10 times, but is not limited thereto.

The extraction method of step 1) includes hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction. Preferably, the extraction may be performed one to five times by an ultrasonic extraction method. The extraction temperature is preferably 10 ° C to 100 ° C and more preferably room temperature, but is not limited thereto. The extraction time is preferably 1 hour to 7 days, more preferably 3 to 7 days, but is not limited thereto.

In addition, the extraction method is a method using an extraction device such as supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction, or a method using an adsorbent resin including XAD and HP-20 or a fermentation or natural fermentation method using microorganisms. Conventional extraction methods in the art may be used, and are preferably heated and extracted at reflux or at room temperature, but are not limited thereto. The extraction number is preferably 1 to 5 times, more preferably 3 times repeated extraction is not limited thereto.

According to one exemplary embodiment, decompression by carbon dioxide, supercritical fluid extraction by high temperature it is possible to prepare a mouse eye extract, the supercritical fluid is generally a liquid having a gas when the critical point at high temperature and high pressure and Supercritical fluids are used for the extraction of fat-soluble substances because of their gaseous properties and chemically similar polarities to nonpolar solvents. Carbon dioxide becomes a supercritical fluid with both liquid and gaseous properties as the pressure and temperature reach a critical point through the operation of a supercritical fluid device, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the fat-soluble substance contained in the sample is extracted from the supercritical carbon dioxide. After extracting the fat-soluble substance, the supercritical carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction container to extract the components that were not extracted with pure supercritical carbon dioxide alone. These cosolvents can be extracted with chloroform, ethanol, One or a mixture of two or more selected from the group consisting of methanol, water, ethyl acetate, hexane and diethyl ether can be used. Most of the extracted samples contain carbon dioxide, which is volatilized into the air at room temperature, and the cosolvent can be removed with a reduced pressure evaporator.

According to an exemplary embodiment, it is possible to produce a mouse eye extract by the ultrasonic extraction method using the energy generated by the ultrasonic vibration. Ultrasonic waves can destroy insoluble substances in the sample in aqueous solvents, and due to the high local temperature generated, the ultrasonic energy increases the kinetic energy of the reactant particles located nearby, thereby obtaining sufficient energy for the reaction. By inducing a high pressure by the impact effect of the to increase the extraction efficiency by increasing the mixing effect of the material and the solvent contained in the sample.

According to one exemplary embodiment, it is possible to prepare a mouse eye extract through a fermentation process. Finely crush the rat bean to about 100 to 500 mesh, and then add 1 to 50 g / L of the conventional microbial culture, and add microorganisms such as yeast or lactic acid in amounts of 10,000 to 100,000 cfu / L. The culture temperature is incubated in a conventional microbial culture conditions of 30 to 37 ℃ and the pH is 5 to 7 incubated for about 5 to 10 days in aerobic or anaerobic conditions, after which the extract can be obtained through aging and filtration.

Concentration under reduced pressure of step 3) is preferably a vacuum rotary evaporator, but is not limited thereto. In addition, it may be dried after vacuum concentration, and drying is preferably, but not limited to, drying under reduced pressure, vacuum drying, boiling drying, spray drying, room temperature drying, or freeze drying.

The organic solvent of step 4) is preferably normal-hexane, methylene chloride, ethyl acetate or normal-butanol, but is not limited thereto.

The fraction of step 4) is obtained by suspending the rat bean extract in water and then systematically fractionating the mixture into normal-hexane, methylene chloride, ethyl acetate, normal-butanol and water, and then extracting the normal-hexane fraction, methylene chloride fraction, and ethyl acetate fraction. It is preferably one of the normal-butanol fraction or the water fraction, more preferably methylene chloride fraction, but is not limited thereto. The fraction can be obtained by repeating the fractionation process 1 to 5 times, preferably three times from the rat eye extract, preferably concentrated under reduced pressure after the fraction is not limited thereto.

In a specific embodiment of the present invention, the rat eye bean extract is ground to an appropriate size to extract the mouse eye bean and placed in an extraction container and left at room temperature by adding an ethanol aqueous solution, and then filtered with a filter paper and concentrated and freeze-dried the filtrate Can be obtained. In addition, by using a separatory funnel obtained by using a separatory funnel, the fractions of mouse eye extract can be obtained by systematically fractionating into normal-hexane, methylene chloride, ethyl acetate, normal-butanol and water fractions.

In an exemplary embodiment, the composition may be 0.1 to 50% by weight of the rat bean extract based on the total weight of the composition. By including 0.1 to 50% by weight of the rat eye extract, based on the total weight of the composition, exhibits significant preventive, improved and / or therapeutic effects of corneal damage and / or corneal disease, and is excellent in efficiency and economical efficiency. It works. Specifically, the composition is 0.5 to 45% by weight, 0.5 to 40% by weight, 0.5 to 35% by weight, 0.5 to 30% by weight, 0.5 to 25% by weight, 0.5 to 20% by weight, 1.0 to 15 relative to the total weight of the composition By weight or 1.0 to 10% by weight mouse eye extract may be included.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of a medicament.

Carriers, excipients and diluents that may be included in the pharmaceutical compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical compositions of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. .

When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, and the like in the composition of the present invention. (sucrose), lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is based on the condition, weight, age, sex of the patient. The range varies depending on the diet, the rate of excretion, the severity of the disease, the form of the drug, the time of administration, the method of administration, the route of administration and the duration of administration. The daily dose is from 0.0001 mg / kg to 500 mg / kg, preferably from 0.001 mg / kg to 100 mg / kg in the amount when the extract according to the present invention is lyophilized. It can be administered in divided doses.

The health food composition of the present invention is a dairy product, including drinks, meat, sausages, bread, biscuits, rice cakes, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, alcoholic beverages And vitamin complexes, and the like, and the health food includes all of the health foods in a conventional sense.

The mouse eye extract of the present invention may be added to food as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient can be suitably determined according to the purpose of use (prevention or improvement). In general, the amount of the extract in the dietary supplement may be added to 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for health and hygiene or health control purposes, the amount may be below that range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

The health functional beverage composition of the present invention is not particularly limited to other ingredients other than containing rat bean extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. . Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the mouse eye extract of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, such as flavoring agents, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the rat eye extract of the present invention may contain a fruit flesh for the production of natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the mouse eye extract of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

실시예 1. 쥐눈이콩 추출물의 제조Example 1 Preparation of Rat Eye Extract

Rat bean ( Rhynchosia nulubilis ) was powdered, and 2 kg of 70% (v / v) ethanol solvent was added to 3 kg of rat bean powder, followed by ultrasonic extraction for 4 hours, and the extract was filtered. After the filtrate was filtered, 15 L of 70% (v / v) ethanol solvent was further added to the remaining residue, and the method of ultrasonic extraction for 4 hours was further repeated twice to obtain 45 L of the total extract. 45 L of the extract was concentrated under reduced pressure at 35 ° C. to obtain 150 g of ethanol extract (EERN) of mouse eye.

실험예Experimental Example 1. 화학적 각막 손상 유발 동물 모델 1. Animal Model of Chemical Corneal Damage

Experimental animals were male BALB / c rats weighing 18-20 g, 6 weeks of age, and were purified for 7 days after purchase in a laboratory environment (temperature 25 ± 1 ° C., humidity 40 ± 5%). Corneal injury-induced animal models were induced by instillation of 0.2% benzalkonium chloride (BAC, 5 μL) twice daily (9 am, 9 pm) for 14 days, and with PBS in the normal control group. . 3 days after the start of the experiment, the rat bean extract obtained in Example 1 was dissolved in 0.5% carboxymethyl cellulose (CMC) solution, and orally administered daily, avoiding eye drops at a dose of 10-50 mg / kg. It was. The untreated control group (Dry eye: DE) was orally administered 0.5% CMC solution containing no rat bean extract in the corneal injury causing animal model. In addition, rats were sacrificed at 14 days after BAC instillation.

On the other hand, benzalkonium chloride (benzalkonium chloride: BAC) is a preservative of the eye drops have excellent antibacterial action and has the effect of improving the stability and corneal permeability of the drug is most used. However, long-term eye drops are known to cause corneal epithelial cell lysis, cell detachment, morphological changes, as well as various inflammatory lesions such as contact dermatitis, conjunctivitis and rhinitis. In humans, since corneal endothelial cells have little regenerative capacity, the damage caused by BAC becomes more severe when BAC is used for a long time. In particular, when corneal epithelial cells are absent or function is reduced, the use of eye drops containing BAC can cause severe corneal damage.

실험예Experimental Example 2. 각막 분석 실험 및 실험 결과 2. Corneal Assay and Experiment Results

The protective effect against corneal damage of the rat eye extract prepared in Example 1 was tested as follows. Specifically, in order to confirm the effect of rat eye extract in BAC-induced corneal injury in Experimental Example 1, fluorescein staining, tear film stability measurement, hematoxylin & eosin staining, TUNEL analysis of cornea The condition of the cornea was analyzed.

(1) Corneal Fluorescein Staining (Corneal fluorescein staining)

After anesthetizing the experimental animals, the corneal damage was observed by using a fluorescent dye (Fluorescein sodium salt). Fluorescein staining stains the substrate at the site where the corneal epithelial cell is missing. 1 μL of 0.1% sodium fluorescein was dropped into the eye of the experimental animal, and the extent of corneal cell damage was photographed using a slit-lamp.

As a result, as shown in Figures 1a and 1b, when comparing the normal control group and the corneal injury causing group, the corneal damage was generated in the corneal injury causing group was confirmed that the corneal damage is greatly reduced when the rat bean extract administration It was. Accordingly, it could be seen that the rat bean extract inhibits corneal cell damage and has a protective effect against corneal damage.

(2) Tear film Break break-up time, TBUT ) Measure

Tear film destruction time is assessed to the extent of damage to corneal epithelial cells. Tear film destruction time was measured using a fluorescein sodium salt. After 1 μL of 0.1% sodium fluorescein was dropped into the eye of the experimental animal, the time for the disappearance of the fluorescent dye was measured.

As a result, as shown in Figure 2, tear film breakdown time was significantly reduced in the corneal injury group, but tear film breakdown time was increased when the rat eye extract was applied to the cornea when the rat eye extract was treated in the dry eye-induced animal model. By protecting the epithelial cells, it was confirmed that the tear film has an effect of stabilizing.

(3) hematoxylin eosin staining method ( Hematoxylin & Esosin staining)

Corneal tissue was sectioned, fixed in 4% paraformaldehyde (pH 7.4) for 20 minutes and washed with PBS. Then, the corneal tissue on the slide was fixed using acetic acid, the nucleus was stained with hematoxylin, and cytoplasm was stained with eosin. The optical microscope was used to confirm the uniformity of corneal epithelial layer and the degeneration of corneal epithelial cells.

As a result, as shown in FIG. 3, it was confirmed that the corneal surface was uneven (see arrow mark) and degeneration of corneal epithelial cells (see circle mark) occurred in the corneal injury causing group. On the other hand, when the rat eye extract was administered, it was confirmed that the corneal surface was uneven or inhibited degeneration of corneal epithelial cells. In the corneal injury group, the corneal parenchyma was thinned, whereas the rat eye extract was used to suppress corneal parenchyma damage by inhibiting this phenomenon and to reduce corneal scratches caused by BAC. It was confirmed that there is. Accordingly, it was found that the rat eye extract has a protective effect against corneal damage by inhibiting corneal epithelial layer and corneal cell degeneration.

(4) TUNEL Analysis (Terminal deoxynucleotidyl transferase dUTP nick end labeling, TUNEL assay)

TUNEL analysis was performed to confirm corneal epithelial cell death in the corneal injury-induced animal model. Corneal sections were exposed to exposed 3′-OH groups of internucleosomally cleaved DNA within the cell using POD (Roche, Mannheim, Germany), an in situ cell death detection kit. Binding and staining with TdT-mediated dUTP nick end labeling (TUNEL) was used to observe the location and extent of expression of cells in which apoptosis occurred directly.

As a result, as shown in Figures 4a and 4b, corneal epithelial cell death was significantly increased in the corneal injury-induced group, and corneal epithelial cell death was decreased when the rat bean extract was administered. Accordingly, it can be seen that the rat eye extract has an effect of inhibiting corneal epithelial cell death in the corneal epithelial cell layer, thereby reducing corneal damage and protecting the cornea.

Examples of the formulation of the composition according to one aspect of the present specification will be described below, which is also applicable to various other formulations, and is not intended to limit the present invention but is only specifically described.

[제제예 1] 점안겔제의 제조Preparation Example 1 Preparation of Eye Drops

Rat Eye Bean Extract 5mg

Carbopol 934 20 mg

Triethanolamine appropriate amount

Paraoxine Benzoate Methyl 2 mg

Sterile purified water ad. 1 g

Paraoxin benzoic acid methyl was added to sterile purified water, heated to dissolve, and cooled to dissolve the rat eye extract. Carbopol 934 was added thereto, mixed with a high speed stirrer, dispersed, and left to stand to remove air. To this, triethanolamine was added drop by drop while being careful not to let air enter, thereby preparing an eye drop.

[[ 제제예Formulation example 2] 점안액제의 제조 2] Preparation of Eye Drops

5 g of rat bean extract

0.1 g of benzalkonium chloride

5 g sodium chloride

6.2 g of boric acid

1.0 g of tyloxapol

Diluted hydrochloric acid

Sterile purified water ad. 1000 ml

Sodium chloride and boric acid were added to the rat bean extract in order, and then dissolved, and benzalkonium chloride and thyroxapol dissolved in a small amount of sterile purified water were added thereto and stirred. Dilute hydrochloric acid was added to adjust pH. Sterilization was performed using a 0.45 microfilter.

[[ 제제예Formulation example 3] 점안연고제의 제조 3] Preparation of Eye Drop Ointment

Rat Eye Bean Extract 5mg

Sterile purified water 10 mg

Paraoxybenzoic acid methyl 2 mg

100 mg anhydrous lanolin

White petrolatum ad. 1 g

Take a rat eye bean extract and paraoxybenzoic acid methyl in a sterile glass bowl, and add the mouse eye bean extract to dissolve, and then mixed with anhydrous lanolin to make it homogeneous until homogeneous.

[[ 제제예Formulation example 4] 4] 연질캅셀제Soft capsule

Mouse eye extract 150 mg, palm oil 2 mg, palm hardened oil 8 mg, lead 4 mg and lecithin 6 mg were mixed and 400 mg per capsule was prepared according to a conventional method to prepare a soft capsule.

[[ 제제예Formulation example 5] 정제 5] tablets

150 mg of rat eye extract, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate were mixed and 40 mg of 30% ethanol was added to form granules, and dried at 60 ° C. using a tablet press. Tableting.

[[ 제제예Formulation example 6] 과립제 6] Granules

Rat eye bean extract 150 mg, glucose 100 mg, red ginseng extract 50 mg and starch 600 mg were mixed and 30 mg of ethanol was added to form granules, followed by drying at 60 ° C. to form granules and then filled into sachets. The final weight of the content was 1 g.

[[ 제제예Formulation example 7] 드링크제 7] Drinks

150 mg of rat eye extract, 10 g of glucose, 50 mg of red ginseng extract, 2 g of citric acid and 187.8 g of purified water were mixed and filled into bottles. The final dose of the contents was 200 ml.

[[ 제제예Formulation example 8] 건강식품의 제조 8] Manufacture of Health Food

Rat Bean Extract ............... 1000 mg

Vitamin mixtures

Vitamin A Acetate ................ 70 μg

Vitamin E ..................... 1.0 mg

Vitamin B1 ......... 0.13 mg

Vitamin B2 ......... 0.15 mg

Vitamin B6 ......... 0.5 mg

Vitamin B12 ......... 0.2 μg

Vitamin C ...................... 10 mg

Biotin ........................ 10 ㎍

Nicotinamide .............. 1.7 mg

Folic acid ........................... 50 ㎍

Calcium Pantothenate ......... 0.5 mg

Mineral mixture

Ferrous Sulfate ........................ 1.75 mg

Zinc oxide ...... 0.82 mg

Magnesium Carbonate ... 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dibasic calcium phosphate ............... 55 mg

Potassium citrate ..................... 90 mg

Calcium Carbonate ......................... 100 mg

Magnesium Chloride ............... 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

[[ 제제예Formulation example 9] 건강음료의 제조 9] Preparation of Health Beverage

Rat Eye Bean Extract ............. 1000mg

Citric acid ..................... 1000 mg

Oligosaccharide ........................ 100 g

Plum concentrate ..... 2 g

Taurine ..................... 1 g

Add purified water to the whole ....... 900 ml

After mixing the above components in accordance with a conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution was filtered and obtained in a sterilized 2 Ll container, sealed sterilization and refrigerated. Then, it was used to prepare a health beverage composition of the present invention.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the blending ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose. Those skilled in the art to which the present invention pertains will be able to perform various applications and modifications within the scope of the present invention based on the above contents.

As described above, specific portions of the present disclosure have been described in detail, and for those skilled in the art, these specific techniques are merely preferred embodiments, and the scope of the present disclosure is not limited thereto. Will be obvious. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims

Pharmaceutical composition for the prevention or treatment of corneal disease comprising rat eye extract as an active ingredient.
The method of claim 1,

The mouse eye extract is extracted with one or more solvents selected from the group consisting of water and C 1 to C 4 lower alcohol, pharmaceutical composition for preventing or treating corneal disease.
The method of claim 1,

The rat eye extract is an ethanol extract, a pharmaceutical composition for preventing or treating corneal disease.
The method of claim 1,

Wherein the composition is to prevent or treat dry eye or corneal abrasions, pharmaceutical composition for the prevention or treatment of corneal disease.
The method of claim 1,

The composition has a corneal epithelial cell death inhibition and corneal epithelial layer protective effect, a pharmaceutical composition for preventing or treating corneal disease.
Health food composition for improving corneal damage comprising rat eye extract as an active ingredient.
The method of claim 6,

The rat eye extract is extracted with one or more solvents selected from the group consisting of water and lower alcohols of C 1 to C 4 , a health food composition for improving corneal damage.
The method of claim 6,

The rat bean extract is an ethanol extract, corneal damage improving health food composition.
The method of claim 6,

The composition is to improve dry eye or corneal abrasions, health food composition for corneal damage improvement.
The method of claim 6,

The composition has a corneal epithelial cell death inhibition and corneal epithelial layer protective effect, corneal damage improvement health food composition.