Composition for preventing or treating ocular diseases comprising aloe vera gel

Abstract

The present invention relates to a composition for preventing or treating ocular diseases, comprising Aloe vera gel. Specifically, when orally administered, the Aloe vera (A.vera) gel has been confirmed to have an excellent effect of increasing the expression of genes related to the improvement of tear volume, corneal non-uniformity, eye inflammation, dry eye, corneal thickness, and tear film. Accordingly, the Aloe vera gel can be advantageously used as an active ingredient in an oral composition for the prevention, alleviation, or treatment of ocular diseases, including dry eye syndrome.

Description

Composition for preventing or treating eye diseases containing aloe vera gel

[0001] The present invention relates to a composition for preventing or treating eye diseases, including aloe vera gel, and more particularly, to an oral composition for preventing or treating eye diseases, including dry eye, including aloe vera gel as an active ingredient.

[0002] Dry eye syndrome is a common condition affecting up to 20% of adults in Korea. It is not simply a lack of tears, but rather a condition characterized by inflammation of the tears and ocular surface (cornea and conjunctiva), which can cause ocular discomfort, decreased vision, and tear film instability, potentially damaging the ocular surface. It is characterized by ocular pain, an irregular corneal surface, blurred and fluctuating vision, and an increased risk of corneal ulcers. While the onset of dry eye is closely related to age, the prevalence is increasing among younger age groups due to prolonged exposure to dry environments, including increased use of contact lenses, computers, and smart devices.

[0003] Specifically, dry eye disease significantly reduces the lubrication of the eye by reducing the secretion of mucus from the corneal and conjunctival epithelium and mucus-secreting goblet cells. Furthermore, dry eye disease causes damage to the corneal surface, increasing the penetration of fluorescein dye into the cornea.

[0004] Treatment for dry eye syndrome is largely limited to conventional therapies, and even these are often highly ineffective. Artificial tears are currently the primary treatment for dry eye. However, these are typically used to supplement insufficient tear production, requiring frequent eye drops. Eye drops derived from sodium hyaluronate and autologous serum have been developed and used for dry eye patients. Furthermore, synthetic compounds such as rebamipide (OPC-127959) and diquafosol sodium, which stimulate tear and mucus secretion, have been developed and utilized. However, long-term use of these medications can lead to various side effects, such as ocular hyperemia and corneal calcification. Concerns about contamination of eye drops, as well as reports of blindness caused by Pseudomonas aeruginosa infection, necessitate the development of safe and effective dry eye treatments, particularly those that are convenient to administer orally.

[0005] Meanwhile, aloe is a perennial plant belonging to the lily family, a succulent CAM (Crassulacean Acid Metabolism) plant with fleshy leaves, and has been widely used as a wide-ranging and multi-purpose folk remedy for over 3,500 years, and more than 400 species of aloe have been identified, among which the juice or gel of aloe vera is one of the representative health functional foods (especially health functional drinks), and is also one of the most widely used natural materials as pharmaceuticals and cosmetics.

[0006] Accordingly, the present inventors have made efforts to develop a natural material for safe and effective treatment of dry eye syndrome, and as a result, have confirmed that when Aloe vera gel is orally administered to a dry eye syndrome animal model, it has excellent effects of improving tear volume, corneal irregularity, ocular inflammation, dry eye, corneal thickness, and increasing gene expression related to tear film improvement. Accordingly, the present inventors have revealed that the Aloe vera gel can be usefully used as an active ingredient in a composition for preventing, improving, or treating ocular diseases including dry eye syndrome, specifically, a composition for oral administration, thereby leading to the present application.

[0007] [Prior Art Literature]

[0008] [Non-patent literature]

[0009] Stern ME et al., "Dry eye as a mucosal autoimmune disease", Int Rev Immunol, 32:19-41, 2013

[0010] Bernauer W et al., "Corneal calcification following intensified treatment with sodium hyaluronate artificial tears", Br J Ophthalmol, 90:285-8, 2006

[0011] The purpose of the present invention is to provide a composition for preventing, improving or treating eye diseases comprising Aloe vera gel.

[0012] In order to achieve the object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating an ocular disease, comprising Aloe vera gel as an active ingredient; a use of Aloe vera gel for use as a pharmaceutical composition for preventing or treating an ocular disease; a use of Aloe vera gel for preparing a pharmaceutical composition for preventing or treating an ocular disease; and a method for treating an ocular disease, comprising a step of administering Aloe vera gel to a subject.

[0013] In addition, the present invention provides a health functional food composition for preventing or improving an eye disease, comprising aloe vera gel as an active ingredient; a use of aloe vera gel for use as a health functional food composition for preventing or improving an eye disease; a use of aloe vera gel for producing a health functional food composition for preventing or improving an eye disease; and a method for preventing or improving an eye disease, comprising a step of administering aloe vera gel to a subject.

[0014] In the present invention, it was confirmed that the Aloe vera (A. vera) gel was orally administered to an animal model of dry eye syndrome and had excellent effects in improving tear quantity, corneal irregularity, eye inflammation, dry eye, corneal thickness, and increasing gene expression related to tear film improvement. Therefore, the Aloe vera gel can be usefully used as an active ingredient of an oral composition for preventing, improving, or treating ocular diseases including dry eye syndrome.

[0015] Figure 1 is a schematic diagram illustrating the production of a dry eye animal model and an administration method according to an embodiment of the present invention using an aloe vera gel administration method.

[0016] Figure 2 is a diagram showing the effect of improving tear volume according to the method of administering aloe vera gel in an animal model of dry eye syndrome according to one embodiment of the present invention.

[0017] FIG. 3 is a diagram showing the effect of improving corneal unevenness according to the method of administering aloe vera gel in an animal model of dry eye syndrome according to one embodiment of the present invention.

[0018] Figure 4 is a diagram showing the effect of improving eye inflammation according to the method of administering aloe vera gel in an animal model of dry eye syndrome according to one embodiment of the present invention.

[0019] FIG. 5 is a diagram showing the effect of aloe vera gel administration on the expression of moisture-related markers in the tear glands in an animal model of dry eye syndrome according to one embodiment of the present invention.

[0020] Figure 6 is a schematic diagram illustrating the production of a dry eye animal model according to one embodiment of the present invention and the administration method according to the dosage of aloe vera gel.

[0021] Figure 7 is a diagram showing the effect of improving tear volume according to the dosage of aloe vera gel administered in a dry eye animal model according to one embodiment of the present invention.

[0022] Figure 8 is a diagram showing the effect of improving corneal unevenness according to the dosage of aloe vera gel administered in a dry eye animal model according to one embodiment of the present invention.

[0023] Figure 9 is a diagram showing the effect of improving eye inflammation according to the dosage of aloe vera gel administered in a dry eye animal model according to one embodiment of the present invention.

[0024] FIG. 10 is a diagram showing the effect of increasing the expression of mucin genes related to the tear film according to the administered dose of aloe vera gel in an animal model of dry eye syndrome according to one embodiment of the present invention.

[0025] Figure 11 is a diagram showing the effect of improving corneal thickness according to the administered dose of aloe vera gel in a dry eye animal model according to one embodiment of the present invention.

[0026] Hereinafter, the present invention will be described in more detail.

[0027] The term "prevention" as used herein refers to inhibiting the occurrence of a disease or condition in an individual who has not been diagnosed with the disease or condition but is prone to such disease or condition. The term "treatment" as used herein refers to (i) inhibiting the development of a disease or condition; (ii) alleviating the disease or condition; and (iii) eliminating the disease or condition.

[0028] The present invention provides a pharmaceutical composition for preventing or treating an ocular disease, comprising Aloe vera gel as an active ingredient; a use of Aloe vera gel for use as a pharmaceutical composition for preventing or treating an ocular disease; a use of Aloe vera gel for preparing a pharmaceutical composition for preventing or treating an ocular disease; and a method for treating an ocular disease, comprising a step of administering Aloe vera gel to a subject.

[0029] In the present invention, the effective ingredient, aloe vera gel, is obtained by removing the peel of a raw aloe vera leaf and separating only the gel inside.

[0030] In the present invention, the aloe vera gel has the following properties and can prevent or treat eye diseases through the following properties:

[0031] 1) Improved tear volume;

[0032] 2) Improvement of corneal irregularity;

[0033] 3) Reduced secretion or expression of inflammatory factors;

[0034] 4) Increased expression of genes associated with tear film improvement; and

[0035] 5) Improvement of corneal thickness.

[0036] Specifically, the aloe vera gel can improve tear production by increasing tear volume.

[0037] In addition, the aloe vera gel can improve eye inflammation by reducing the secretion or expression of inflammation-related factors, examples of which include HMGB-1, p-JNK, p-ERK, and p-NF-κB.

[0038] In addition, the aloe vera gel can improve dry eye by increasing the expression of genes related to tear film improvement, and examples of genes related to tear film improvement include aquaporin genes, specifically AQP-1, and mucin genes, specifically MUC1 and MUC2.

[0039] In the present invention, the ocular disease may be an ocular surface disease or an intraocular disease. Examples of the ocular surface disease include dry eye, corneal epithelial damage, keratitis, conjunctivitis, or keratoconjunctivitis, and examples of the intraocular disease include, but are not limited to, endophthalmitis, uveitis, or macular degeneration.

[0040] Additionally, the dry eye may be a dry eye-like condition including, but not limited to, amenorrhea, xeroderma, Sjogren's syndrome, keratoconjunctivitis sicca, Stevens-Johnson syndrome, ocular pseudobulbar bleb, post-ophthalmic dry eye, dry eye secondary to allergic conjunctivitis, tear reduction in VDT ​​(Visual Display Terminal) workers, or tear reduction without any systemic symptoms caused by dry rooms due to air conditioning units.

[0041] In the present invention, the composition can be administered orally.

[0042] In a specific example, the present inventors confirmed that when aloe vera gel is orally administered to an animal model of dry eye syndrome, it has superior effects of improving tear volume, corneal irregularity, ocular inflammation, and increasing aquaporin gene expression related to tear film improvement, compared to when it is administered by eye drops. In addition, when aloe vera gel is orally administered to an animal model of dry eye syndrome, it has additionally superior effects of improving dry eye, corneal thickness, and tear film improvement, as well as increasing mucin gene expression related to tear film improvement. Therefore, aloe vera gel can be usefully used as an active ingredient in a pharmaceutical composition for preventing or treating ocular diseases including dry eye syndrome, particularly in a pharmaceutical composition for oral administration for preventing or treating ocular diseases.

[0043] The pharmaceutical composition according to the present invention may further comprise a pharmaceutically acceptable carrier or additive. As used herein, “pharmaceutically acceptable” means physiologically acceptable and does not typically cause allergic reactions such as gastrointestinal upset or dizziness or similar reactions when administered to humans. Examples of the additives include excipients, disintegrants, binders, lubricants, wetting agents, dispersants, stabilizers, etc. Examples of the excipients include lactose, mannitol, isomalt, microcrystalline cellulose, silicified microcrystalline cellulose, powdered cellulose, etc. Examples of the disintegrant include low-substituted hydroxypropyl cellulose, crospovidone, sodium starch glycolate, croscarmellose sodium, starch, etc. Examples of the binder include hydroxypropyl cellulose, hypromellose, povidone, copovidone, pregelatinized starch, etc. Examples of lubricants include stearic acid, magnesium stearate, and sodium stearyl fumarate. Examples of humectants include polyoxyethylene sorbitan fatty acid ester derivatives, poloxamers, and polyoxyethylene castor oil derivatives. Examples of dispersants include hypromellose, hydroxypropylcellulose, povidone, copovidone, sodium carboxymethylcellulose, and methylcellulose. Examples of stabilizers include citric acid, fumaric acid, and succinic acid. In addition, the pharmaceutical composition of the present invention may additionally include anticoagulants, fragrances, emulsifiers, and preservatives.

[0044] Furthermore, the pharmaceutical composition of the present invention can be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to a mammal. The pharmaceutical formulation may be a powder, granule, tablet, suspension, emulsion, syrup, aerosol, or soft or hard gelatin capsule.

[0045] The dosage of the active ingredient used in the composition according to the present invention depends on the subject being treated, the severity of the disease or condition, the rate of administration, and the judgment of the prescribing physician. The active ingredient can be administered orally to mammals, including humans, in an amount of 0.05 to 100 mg / kg (body weight) per day, preferably 0.5 to 50 mg / kg (body weight). In some cases, dosages lower than the above-mentioned range may be more suitable, and higher dosages may be used without causing harmful side effects, and in the case of higher dosages, they are divided into several smaller doses throughout the day.

[0046] In addition, the present invention provides a health functional food composition for preventing or improving an eye disease, comprising Aloe vera gel as an active ingredient; a use of Aloe vera gel for use as a health functional food composition for preventing or improving an eye disease; a use of Aloe vera gel for producing a health functional food composition for preventing or improving an eye disease; and a method for preventing or improving an eye disease, comprising a step of administering Aloe vera gel to a subject.

[0047] In the present invention, the contents of the aloe vera gel, properties and diseases are the same as those described in the pharmaceutical composition for preventing or treating eye diseases containing aloe vera gel as an active ingredient, so the detailed description is based on the above contents, and only the unique composition of the health functional food composition is described below.

[0048] In a specific example, the inventors of the present invention confirmed that when aloe vera gel is orally administered to an animal model of dry eye syndrome, it has superior effects of improving tear volume, corneal irregularity, ocular inflammation, and increasing aquaporin gene expression related to tear film improvement, compared to when aloe vera gel is administered by eye drops. In addition, when aloe vera gel is orally administered to an animal model of dry eye syndrome, it has additionally superior effects of improving dry eye, corneal thickness, and tear film improvement, and therefore, the aloe vera gel can be usefully used as an active ingredient of a health functional food composition for preventing or improving ocular diseases including dry eye syndrome.

[0049] The health functional food composition of the present invention may be manufactured in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, liquid or liquid, flake, paste, gel, jelly, bar, film, and beverage, but is not limited thereto. In addition, the aloe vera gel includes a dilution, a concentrate, a dried product obtained by drying a dilution or concentrate, a modified product, a purified product, or a combination thereof.

[0050] The above health functional food composition is not particularly limited as long as it can be consumed to prevent or improve eye diseases. When the health functional food composition of the present invention is used as a food additive, the health functional food composition can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The active ingredient can be used appropriately depending on its intended use (prevention or improvement). Generally, when manufacturing a food or beverage, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less. However, in the case of long-term intake for the purpose of health control, the amount can be below the above range, and since there is no problem in terms of safety, the active ingredient can be used in an amount greater than the above range. There is no particular limitation on the type of the food. Examples of foods to which the above health functional food composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the conventional sense. In addition, the health functional food composition of the present invention can be manufactured into a food, particularly a functional food.

[0051] The functional food of the present invention includes ingredients commonly added during food manufacturing, such as proteins, carbohydrates, fats, nutrients, and seasonings. For example, when manufactured as a drink, it may include natural carbohydrates or flavoring agents as additional ingredients in addition to the active ingredients. The natural carbohydrates are preferably monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, etc.), or sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). The flavoring agents may be natural flavoring agents (e.g., thaumatin, stevia extract, etc.) or synthetic flavoring agents (e.g., saccharin, aspartame, etc.). In addition to the above health functional food composition, it may further contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

[0052] Hereinafter, the present invention will be described in detail through examples and experimental examples.

[0053] However, the following examples and experimental examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples and experimental examples.

[0054] <Example 1> Preparation of Aloe vera gel for oral administration

[0055] Aloe vera (A. vera) raw plants were washed and the aloe gel was separated physically or chemically. Thereafter, anthraquinone was removed with activated carbon, concentrated, and dried to obtain a 200:1 aloe vera gel powder raw material. Then, 160 mg, 250 mg, 320 mg, 500 mg, and 640 mg of the aloe vera gel raw material were each mixed in 10 mL of triple-distilled water and homogenized to prepare an aloe vera gel for oral administration (hereinafter referred to as 'AO').

[0056] <Comparative Example 1> Preparation of aloe vera gel for eye drops

[0057] 5 mg of the aloe vera gel raw material obtained in the above <Example 1> was mixed with 10 mL of PBS (Phosphate-Buffered Saline) and homogenized to prepare an aloe vera gel for eye drops (hereinafter referred to as 'AE').

[0058] <Experimental Example 1> Evaluation of the improvement effect of aloe vera gel administration method in an animal model of dry eye syndrome.

[0059] <1-1> Creation of an animal model of dry eye syndrome and administration of aloe vera gel

[0060] To determine the effect of aloe vera gel on improving ocular disease according to the method of administration, an animal model of dry eye was created, and aloe vera gel was administered orally or as eye drops.

[0061] Specifically, C57BL / 6 mice (7 weeks old, female) were obtained from Samtaco BIO KOREA Co., Ltd. The C57BL / 6 mice (7 weeks old, female) were provided with food and water ad libitum. The experimental animals were maintained at 22±2℃, relative humidity 55±5%, and a 12-h light / dark cycle, and the experiment was conducted with the approval of the Animal Experiment Ethics Committee of Chonbuk National University (NON2023-080).

[0062] As shown in the schematic diagram in Fig. 1, the experimental animals were given a one-week adaptation period, and then divided into a general control group (a group not induced with dry eye disease, Control, Con), a dry eye disease control group (DED), an AG eye drop group (AE 0.05%), a low-concentration AG oral administration group (AO 250 mg / kg), and a high-concentration AG oral administration group (AO 500 mg / kg) as shown in [Table 1] below.

[0063] Next, to induce dry eye syndrome in the remaining groups, excluding the general control group (Con), 0.2% benzalkonium chloride (BAC) was instilled into both eyes at 5 μL every 12 hours / day (9 am, 9 pm) for 2 weeks. Two weeks after inducing dry eye syndrome, the normal control group (Con) and the dry eye control group (DED) were orally administered PBS for two weeks, the AG eye drop group (AE 0.05%) was orally administered AE containing 0.05% of the aloe gel raw material prepared in <Comparative Example 1>, and the low-concentration AG oral administration group (AO 250 mg / kg) and high-concentration AG oral administration group (AO 500 mg / kg) were orally administered AO containing 250 mg / kg and 500 mg / kg of the aloe gel raw material prepared in <Example 1>, respectively.

[0064] Group Test Method ControlPBS Oral AdministrationDED0.2% BAC Eye Drops + PBS Oral AdministrationAE0.05%0.2% BAC Eye Drops + 0.05% AE Eye DropsAO250mg / kg0.2% BAC Eye Drops + 250mg / kg AO Oral AdministrationAO500mg / kg0.2% BAC Eye Drops + 500mg / kg AO Oral Administration

[0065] <1-2> Evaluation of the effect of improving tear volume according to the method of administering aloe vera gel

[0066] In order to determine the effect of aloe vera gel on improving eye diseases according to the method of administration, AE was administered as eye drops and AO was administered orally to the experimental animals of the above experimental example <1-1>, and then the change in tear volume was evaluated.

[0067] Specifically, immediately after the induction of dry eye syndrome in the experimental animals of the above experimental example <1-1>, the tear volume was measured every 8 o'clock on days 0, 7, and 14 using SMTube (Strip meniscometry tube, Echo Electricity, Japan) according to the manufacturer's procedure.

[0068] As a result, as shown in Fig. 2, on the 0th day of aloe vera gel administration, dry eye was induced in all groups except the general control group (Con), and tear volume decreased. On the 7th day of aloe vera gel administration, the tear volume of the AO 500 mg / kg group was similar to that of the general control group, and the AO 250 mg / kg group tended to increase compared to the dry eye control group (DED). On the other hand, the AG eye drop group (AGE 0.05%) showed a similar tear volume to that of the dry eye control group (DED). On the 14th day of aloe vera gel administration, the tear volume increased in the AG oral administration groups (AO 250 mg / kg and AO 500 mg / kg) to a similar value to that of the general control group. On the other hand, the AG eye drop group (AE 0.05%) showed a slight increase in tear volume compared to the dry eye control group (DED), but there was no significant difference.

[0069] These results demonstrate that oral administration of aloe vera gel can restore tear volume. Furthermore, oral administration of aloe vera gel significantly improves tear volume compared to topical administration.

[0070] <1-3> Evaluation of the effect of aloe vera gel administration on improving corneal irregularity

[0071] In order to determine the effect of aloe vera gel on improving eye diseases according to the method of administration, AE was administered as eye drops and AO was administered orally to the experimental animals of the above experimental example <1-1>, and then the unevenness of the cornea was evaluated.

[0072] Specifically, the experimental animals of the above experimental example <1-1> were anesthetized with anesthetic (Alfaxan, 0.25-0.5 mL / kg) in the 5th week of the experiment and their movements were corrected. Then, the unevenness of the cornea was analyzed after ring-shaped light irradiation using a stereo microscope (Stereo microscope, SMART ZOOM5, Carl Zeiss, Germany) in the Chonbuk National University Joint Experimental Practice Center.

[0073] As a result, as shown in Fig. 3, in the dry eye control group (DED), the ring structure (white ring-shaped light) of the eye lost its shape overall, and it was confirmed that the ring structure in each part of the AG eye drop group (AE 0.05%) also became blurred. On the other hand, it was confirmed that the AG oral administration group (AO 250 mg / kg and AO 500 mg / kg) maintained the circular ring structure well, similar to the normal control group (Con).

[0074] The above results show that oral administration of aloe vera is more effective in improving corneal irregularity than eye drops.

[0075] <1-4> Evaluation of the effect of aloe vera gel on improving eye inflammation according to the method of administration

[0076] HMGB-1 is known as a marker that induces inflammatory responses and cell damage in dry eye, and p-JNK, p-ERK, and p-NFkB are known as inflammation-related signaling proteins. Therefore, in order to investigate the ocular disease improvement effect according to the administration method of aloe vera gel, AE was administered instilled into the experimental animals of Experimental Example <1-1> and AO was administered orally, and then the expression of HMGB-1, p-JNK, p-ERK, and p-NFkB proteins in the ocular tissue was analyzed.

[0077] Specifically, the experimental animals of Experimental Example <1-1> were sacrificed after the experiment was over, and the eyeballs were incised and the ocular tissues were harvested in ice-cold 1X PBS. Then, they were lysed in RIPA buffer (Thermo Fisher Scientific; Waltham, MA, USA) containing protease inhibitors (Gendepot; Katy, TX, USA). The protein concentration of the ocular tissue lysate was quantified using a BCA protein assay kit (Thermo Fisher Scientific). The lysate containing 20 μg of protein was mixed with SDS sample buffer and heated at 99°C for 5 minutes. After proteins were separated by SDS-PAGE at 200 V for 90 min, they were transferred to nitrocellulose membranes and blotted with anti-HMGB-1 antibody (1:100 dilution, Santa Cruz), anti-p-JNK antibody (1:500 dilution, Cell Signaling), and anti-p-ERK antibody (1:1000 dilution, Cell Signaling). Subsequently, goat anti-mouse HRP and goat anti-rabbit HRP antibodies (1:20000 dilution, Thermo Fisher Scientific) were used. Protein bands were detected using ECL reagent (GE Healthcare; Marlborough, MA, USA). Film images were captured with a Bio 5000 scanner (Microtek, Hsinchu, Taiwan). Protein band sizes were quantified using ImageJ open source software (version 1.45s, National Institute of Health, NIH; Bethesda, MD, USA).

[0078] As a result, as shown in Fig. 4, in the dry eye control group (DED), HMGB-1 protein expression was significantly increased compared to the normal control group (CON), whereas HMGB-1 protein expression was decreased in the AG administration group, and particularly, in the high-dose AG oral administration group (AO 500 mg / kg), HMGB-1 protein expression was decreased compared to the normal control group (CON). In addition, it was confirmed that the expression of inflammation-related signaling proteins p-JNK, p-ERK, and p-NFkB was significantly increased in the dry eye control group (DED) compared to the normal control group (CON), and was decreased by AG administration. In particular, it was confirmed that the expression of inflammation-related signaling proteins p-JNK, p-ERK, and p-NFkB was more significantly decreased in the AG oral administration group (AO 250 mg / kg and AO 500 mg / kg) than in the AG eye drop group (AE 0.05%).

[0079] <1-5> Evaluation of tear gland moisture secretion markers according to the method of aloe vera gel administration

[0080] In order to determine the effect of aloe vera gel on improving eye diseases according to the method of administration, AE was administered by eye drop and AO was administered orally to the experimental animals of the above experimental example <1-1>, and then the protein expression of AQP-1, a water secretion marker, was analyzed in the tear gland tissue.

[0081] Specifically, the experimental animals of the above Experimental Example <1-1> were sacrificed after the experiment was completed, the eyeballs were incised, and the lacrimal gland tissues were harvested in ice-cooled 1X PBS. Then, the protein expression of AQP1 was confirmed using the same method as described in the above Experimental Example <1-5> using an anti-AQP1 antibody (1:500 dilution, Abclonal).

[0082] As a result, as shown in Fig. 5, in the dry eye control group (DED), AQP-1 protein expression was lower than in the normal control group (CON), whereas in the AG oral administration group (AO 250 mg / kg and AO 500 mg / kg), AQP-1 protein expression was confirmed to be similar to that in the normal control group (CON). Meanwhile, in the AG eye drop group (AE 0.05%), AQP-1 protein was found to be low, similar to that in the dry eye control group (DED).

[0083] <Experimental Example 2> Evaluation of the improvement effect according to the dosage of aloe vera gel administered in an animal model of dry eye syndrome.

[0084] <2-1> Creation of an animal model of dry eye syndrome and administration of aloe vera gel

[0085] To determine the effect of aloe vera gel on improving ocular disease according to the dosage, an animal model of dry eye was created and aloe vera gel was orally administered at different dosages.

[0086] Specifically, C57BL / 6 mice (7 weeks old, female) were obtained from Samtaco BIO KOREA Co., Ltd. The C57BL / 6 mice (7 weeks old, female) were provided with food and water ad libitum. The experimental animals were maintained at 22±2℃, relative humidity 55±5%, and a 12-h light / dark cycle, and the experiment was conducted with the approval of the Animal Experiment Ethics Committee of Chonbuk National University (NON2023-080).

[0087] As shown in the schematic diagram of Fig. 6, after a one-week adaptation period, the experimental animals were divided into a general control group (a group not induced with dry eye syndrome, Control, Con), a dry eye disease control group (DED), an omega-3 oral administration group (ω-3 180 mg / kg), a low-concentration AG oral administration group (AO 160 mg / kg), a medium-concentration AG oral administration group (AO 320 mg / kg), and a high-concentration AG oral administration group (AO 640 mg / kg), as shown in [Table 2] below.

[0088] Next, to induce dry eye syndrome in the remaining groups, excluding the general control group (Con), 0.2% benzalkonium chloride (BAC) was instilled into both eyes at 5 μL every 12 hours / day (9 am, 9 pm) for 2 weeks. Two weeks after inducing dry eye syndrome, the normal control group (Con) and dry eye control group (DED) were orally administered PBS for two weeks, the omega-3 oral administration group (ω-3) was orally administered 180 mg / kg of omega-3 (Lot No. 2023000159) provided by Univera, and the low-concentration AG oral administration group (AO 160 mg / kg), medium-concentration AG oral administration group (AO 320 mg / kg), and high-concentration AG oral administration group (AO 640 mg / kg) were orally administered AO containing 160 mg / kg, 320 mg / kg, and 640 mg / kg of the aloe gel raw material manufactured in <Example 1>, respectively.

[0089] Group Test Method ControlPBS Oral AdministrationDED0.2% BAC Eye Drops + PBS Oral Administrationω-30.2% BAC Eye Drops + 180mg / kg Omega-3 Oral AdministrationAO 160mg / kg0.2% BAC Eye Drops + 160mg / kg AO Oral AdministrationAO 320mg / kg0.2% BAC Eye Drops + 320mg / kg AO Oral AdministrationAO 640mg / kg0.2% BAC Eye Drops + 640mg / kg AO Oral Administration

[0090] <2-2> Evaluation of the effect of improving tear volume according to the dosage of aloe vera gel administered

[0091] In order to determine the effect of aloe vera gel on improving eye diseases according to the dosage, AE was administered to the experimental animals of the above experimental example <2-1> by eye drops and AO was administered orally, and then the change in tear volume was evaluated.

[0092] Specifically, the tear amount was measured using the experimental animal of the above experimental example <2-1> using the same method as described in the above experimental example <1-2>.

[0093] As a result, as shown in Fig. 7, both the omega-3 oral administration group (ω-3) and the AG oral administration group (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg) showed a tendency for tear secretion to significantly increase compared to the dry eye control group (DED) from around the 7th day of the experiment, and the same tendency was observed on the 14th day.

[0094] The above results show that oral administration of aloe vera gel has an excellent effect in improving tear volume.

[0095] <2-3> Evaluation of the effect of improving corneal irregularity according to the dosage of aloe vera gel administered

[0096] In order to determine the effect of aloe vera gel on improving eye diseases according to the dosage, AE was administered as eye drops and AO was administered orally to the experimental animals of the above experimental example <2-1>, and then the unevenness of the cornea was evaluated.

[0097] Specifically, the corneal unevenness was analyzed using the experimental animal of the above experimental example <2-1> using the same method as described in the above experimental example <1-3>.

[0098] As a result, as shown in Fig. 8, in the dry eye control group (DED), the ring structure (white ring-shaped light) of the eye lost its shape overall, and in the omega-3 oral administration group (ω-3), the ring structure was clearer than in the dry eye control group (DED), but it was confirmed to have a slightly cloudy appearance. On the other hand, in the AG oral administration group (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg), it was confirmed that the ring structure was maintained in a circular shape and improved in a concentration-dependent manner.

[0099] The above results show that oral administration of aloe vera gel has an excellent effect in improving corneal irregularity.

[0100] <2-4> Evaluation of the effect of improving eye inflammation according to the dosage of aloe vera gel administered

[0101] In order to determine the effect of aloe vera gel on improving eye diseases according to the dosage, AE was administered by eye drop and AO was administered orally to the experimental animals of the above experimental example <2-1>, and then the protein expression of HMG-1, p-JNK, p-ERK, and p-NFkB in the eye tissue was analyzed.

[0102] Specifically, the protein expression of HMG-1, p-JNK, p-ERK, and p-NFkB was confirmed using the experimental animal of the above experimental example <2-1> using the same method as described in the above experimental example <1-3>.

[0103] As a result, as shown in Fig. 9, in the dry eye control group (DED), HMGB-1 protein expression increased compared to the normal control group (CON), whereas in the omega-3 oral administration group (ω-3), it decreased compared to the dry eye control group (DED). In addition, HMGB-1 protein expression also decreased in a concentration-dependent manner in the AG oral administration groups (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg).

[0104] As for the inflammation-related signaling proteins, p-JNK, p-ERK, and p-NFkB proteins, similar to HMGB-1 protein, were confirmed to have a decreased expression in the omega-3 oral administration group (ω-3) and the AG oral administration group (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg) compared to the dry eye control group (DED). In particular, the AG oral administration group (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg) showed an expression level that was as low as or lower than the omega-3 oral administration group (ω-3).

[0105] The above results show that oral administration of aloe vera gel is effective in improving eye inflammation.

[0106] <2-5> Evaluation of mucus-related markers according to the administered dose of aloe vera gel

[0107] In order to determine the effect of improving ocular diseases according to the dosage of aloe vera gel, AE was administered by eye drop and AO was administered orally to the experimental animals of the above experimental example <2-1>, and then the mRNA expression of MUC1 and MUC2, which are glycoproteins involved in gel formation as mucus-related markers in ocular tissue, was analyzed.

[0108] Specifically, the experimental animals of Experimental Example <2-1> were sacrificed after the experiment was completed, and the eyeballs were dissected and the ocular tissues were harvested in ice-cold 1X PBS. Next, the harvested ocular tissues were treated with TRIzol reagent to isolate total RNA, and cDNA was synthesized from the total RNA. The cDNA was subjected to quantitative real-time PCR using primers for MUC1 and MUC2 and power SYBR Green PCR Master Mix according to the manufacturer's procedure. The expression of each gene was normalized to the expression of GAPDH.

[0109] As a result, as shown in Fig. 10, in the dry eye control group (DED), the mRNA expression of MUC1 and MUC2 decreased compared to the normal control group (CON), whereas in the AG oral administration group (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg), the expression was confirmed to increase in a concentration-dependent manner compared to the dry eye control group (DED). In particular, the omega-3 oral administration group (ω-3) showed a decrease in the mRNA expression of MUC1 compared to the dry eye control group (DED).

[0110] The above results show that oral administration of aloe vera gel has an excellent effect in improving dry eye.

[0111] <2-6> Evaluation of corneal thickness improvement effect according to the dosage of aloe vera gel administered

[0112] In order to determine the effect of aloe vera gel on improving eye diseases according to the dosage, AE was administered to the experimental animals of the above experimental example <2-1> by eye drops and AO was administered orally, and then tissue staining was performed on the cornea and conjunctiva tissue.

[0113] Specifically, the experimental animals of the above Experimental Example <2-1> were sacrificed after the experiment was completed, and the eyes were incised to harvest the corneal and conjunctival tissues in ice-cold 1X PBS. Then, the corneal and conjunctival tissues were fixed in 10% formalin solution, made into paraffin blocks, and H&E staining (KP&T, Cheongju, Korea) was performed. The tissue slides were photographed using a microscope in the laboratory.

[0114] As a result, as shown in Fig. 11, in the dry eye control group (DED), corneal thickness was significantly reduced compared to the normal control group (CON), whereas in the omega-3 oral administration group (ω-3) and the AG oral administration group (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg), corneal thickness was improved. In particular, it was confirmed that the AG oral administration group (AO 160 mg / kg, AO 320 mg / kg, AO 640 mg / kg) had a superior effect on improving corneal thickness compared to the omega-3 oral administration group (ω-3).

[0115] The above results show that oral administration of aloe vera gel has an excellent effect in improving corneal thickness.

[0116] In the present invention, it was confirmed that the Aloe vera (A. vera) gel was orally administered to an animal model of dry eye syndrome and had excellent effects in improving tear quantity, corneal irregularity, eye inflammation, dry eye, corneal thickness, and increasing gene expression related to tear film improvement. Therefore, the Aloe vera gel can be usefully used as an active ingredient of an oral composition for preventing, improving, or treating ocular diseases including dry eye syndrome.

Claims

1. A pharmaceutical composition for preventing or treating eye diseases, comprising aloe vera gel as an active ingredient.

2. A pharmaceutical composition for preventing or treating ocular diseases, wherein the aloe vera gel improves tear volume, corneal irregularity, or corneal thickness in the first paragraph.

3. A pharmaceutical composition for preventing or treating ocular diseases, wherein the aloe vera gel reduces the secretion or expression of inflammatory factors in the first paragraph.

4. A pharmaceutical composition for preventing or treating ocular diseases, wherein the inflammation-related factor in paragraph 3 is at least one selected from the group consisting of HMGB-1, p-JNK, p-ERK, and p-NF-κB.

5. A pharmaceutical composition for preventing or treating ocular diseases, wherein the aloe vera gel in claim 1 increases the expression of genes related to tear film improvement.

6. A pharmaceutical composition for preventing or treating ocular diseases, wherein the gene related to tear film improvement in paragraph 5 is an aquaporin gene or a mucin gene.

7. A pharmaceutical composition for preventing or treating ocular diseases, wherein the aquaporin gene is AQP-1 and the mucin gene is MUC1 or MUC2 in claim 6.

8. A pharmaceutical composition for preventing or treating an ocular disease, wherein the ocular disease in claim 1 is an ocular surface disease selected from the group consisting of dry eye, corneal epithelial damage, keratitis, conjunctivitis, and keratoconjunctivitis; or an intraocular disease selected from the group consisting of endophthalmitis, uveitis, and macular degeneration.

9. A pharmaceutical composition for preventing or treating an ocular disease, wherein the dry eye is any one selected from the group consisting of dry eye-like conditions including amenorrhea, dry cornea, Sjogren's syndrome, keratoconjunctivitis sicca, Steven-Johnson syndrome, ocular pseudobulbar bleb, dry eye after ophthalmic surgery, dry eye accompanying allergic conjunctivitis, tear reduction in VDT ​​(Visual Display Terminal) workers, and tear reduction without any systemic symptoms caused by a dry room due to an air conditioning device.

10. A pharmaceutical composition for preventing or treating ocular diseases, wherein the composition is administered orally in accordance with paragraph 1.

11. A health functional food composition for preventing or improving eye diseases, containing aloe vera gel as an active ingredient.

12. A health functional food composition for preventing or improving eye diseases, wherein the composition is manufactured in any one of the following forms: powder, granules, pills, tablets, capsules, candy, syrup, liquid or liquid, flakes, paste, gel, jelly, bar, film, and beverage.

13. Use of aloe vera gel for preparing a pharmaceutical composition for preventing or treating eye diseases.

14. Use of aloe vera gel for manufacturing a health functional food composition for preventing or improving eye diseases.

15. A method for treating an eye disease, comprising administering aloe vera gel to a subject.

16. A method for preventing or improving an eye disease, comprising administering aloe vera gel to a subject.

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