KR102284682B1 - Composition for prevention or treatment of retinal diseases comprising compounds isolated from Centella asiatica extract - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating retinal diseases, such as glaucoma and macular degeneration, comprising a compound isolated from Centella asiatica extract as an active ingredient. It can be usefully used as a composition for preventing and improving or treating retinal diseases such as glaucoma and macular degeneration for eye health by increasing the survival rate of and protecting retinal cells from oxidative damage.

Description

Composition for prevention or treatment of retinal diseases comprising compounds isolated from Centella asiatica extract as an active ingredient {Composition for prevention or treatment of retinal diseases comprising compounds isolated from Centella asiatica extract}

The present specification relates to a composition for improving, preventing or treating retinal diseases comprising asiaticoside or asiatic acid, which is a compound isolated from Centella asiatica extract, as an active ingredient.

Glaucoma is a disease in which the optic nerve is damaged due to an increase in intraocular pressure or blood circulation abnormality in the optic nerve. The common cause is an increase in intraocular pressure due to circulatory disturbance of the aqueous humor. As the age of 40 years or older, the aqueous humor outlet becomes narrower, and the amount of aqueous humor produced increases, resulting in increased intraocular pressure, which causes pressure and damage to the optic nerve. . In addition, although intraocular pressure is normal, glaucoma develops in some cases. In particular, in Korea and Japan, there are more glaucoma patients with normal intraocular pressure (about 70 to 80% of glaucoma patients), and the cause is known as optic nerve blood circulation disorder. In fact, it is known that there are many cases of symptoms of peripheral blood circulation disorders caused by metabolic diseases in patients in the range of normal intraocular pressure. Ultimately, glaucoma leads to the death of cells such as retinal ganglion cells (Almasieh et al., 2012), and substances that can prevent and treat the death of these cells can be used as food and therapeutic agents to inhibit the onset and progression of glaucoma. .

The macula is the part of the retina of the eye that receives the most clear and accurate light due to the concentration of photoreceptors, and plays a very important role in vision. This is called macular degeneration. Macular degeneration is a disease that occurs due to the gradual degeneration of retinal epithelial cells, retina, and choroidal capillary layer. Macular degeneration in the early stage is caused by extracellular deposition called drusen between Bruch's membrane and retinal epithelial cells. It is characterized by the observation of abnormal retinal epithelial cells formed. Also, in the later stage, it is largely divided into exudative (wet) macular degeneration and atrophic (dry) macular degeneration. In both types of macular degeneration, the photoreceptor cells in the macula are gradually destroyed, so as time goes by, the function of the macula decreases and central vision deteriorates. will decrease Oxidative stress and inflammation are known to be the main causes of macular degeneration. In particular, if by-products after intracellular metabolism are gradually accumulated without being discharged from the retinal pigment epithelial cells, they are subsequently subjected to an oxidative stress process, which eventually leads to apoptosis. (Sparrow and Cai, 2001). Therefore, prevention of oxidative stress in retinal pigment epithelial cells can reduce the risk of macular degeneration itself.

Glaucoma and macular degeneration are one of the three major causes of blindness along with diabetic retinopathy, and death of optic nerve cells or retinal nerve cells can be said to be the main causes. It is known that the number of large patients is increasing rapidly.

Recently, the development of eye health maintenance medicines and health functional foods such as prevention of oxidative damage to the eyes has been active. In particular, energy improving agents using anthocyanosides derived from wild blueberries as the main ingredient have been developed. herbal medicines have been used for the purpose of maintaining and improving vision since ancient times, but there is no research on how to prevent or treat glaucoma and retinal macular degeneration related to retinal diseases using the main components of Centella asiatica.

Korean Patent No. 10-1700903 Korean Patent Publication No. 10-2018-0080584

Therefore, the present inventors recognize the importance of prevention and treatment for retinal diseases such as glaucoma and macular degeneration, and use various natural materials and main ingredients to provide natural resources that can protect the retinal pigment epithelial cell proliferation efficacy and oxidative damage. was selected. As a result, Centella asiatica extract showed excellent proliferation efficacy in optic nerve cells and retinal cells, inhibited oxidative damage caused by A2E in retinal cells, and greatly increased glucose availability in retinal cells. In particular, asiaticoside and asiatic acid, which are compounds isolated from Centella asiatica extract, had the survival rate of retinal pigment epithelial cells and a cytoprotective effect from oxidative stress, and greatly improved glucose availability in retinal cells. increased. It can be inferred that the recovery effect and protective effect according to cell damage are excellent, so the compound isolated from the Centella asiatica extract of the present invention has a great potential for commercialization as a composition for the prevention, improvement, or treatment of glaucoma and macular degeneration.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing, improving or treating retinal diseases, comprising a compound isolated from Centella asiatica extract.

Another object of the present invention is to provide a health functional food composition for the prevention and improvement of retinal diseases comprising a compound isolated from Centella asiatica extract.

The object of the present invention is not limited to the object mentioned above. The objects of the present invention will become more apparent from the following description, and will be realized by means and combinations thereof described in the claims.

One aspect of the present invention is for protecting retinal cells from oxidative stress, comprising asiaticoside or asiatic acid, a compound isolated from Centella asiatica extract, or a pharmaceutically acceptable salt as an active ingredient. A food composition is provided.

In one aspect of the present invention, asiaticoside or asiatic acid, which is a compound isolated from the Centella asiatica extract, provides a food composition that improves the survival rate of retinal cells by increasing sugar availability.

In one aspect of the present invention, asiaticoside or asiatic acid, which is a compound isolated from the Centella asiatica extract, provides a food composition that prevents retinal cell death from oxidative stress in retinal cells.

In one aspect of the present invention, the food composition provides a food composition for preventing or improving retinal cell damage diseases caused by oxidative stress.

Another aspect of the present invention is to prevent, improve or treat retinal cell damage caused by oxidative stress, comprising asiaticoside or asiatic acid or a pharmaceutically acceptable salt as an active ingredient, which is a compound isolated from Centella asiatica extract. It provides a pharmaceutical composition for.

In another aspect of the present invention, asiaticoside or asiatic acid, which is a compound isolated from the Centella asiatica extract, provides a pharmaceutical composition that improves the survival rate of retinal cells by increasing sugar availability.

In another aspect of the present invention, asiaticoside or asiatic acid, which is a compound isolated from the Centella asiatica extract, provides a pharmaceutical composition that prevents retinal cell death from oxidative stress.

In another aspect of the present invention, the disease is macular degeneration or glaucoma, to provide a pharmaceutical composition.

In one or another aspect of the present invention, the compound provides a composition isolated from Centella asiatica extracted with water, a C1-6 lower alcohol, or a mixed solvent thereof.

In one or another aspect of the present invention, there is provided a composition wherein the asiaticoside or asiatic acid, which is a compound isolated from the Centella asiatica extract, is administered at a dosage of 0.004 to 40 mg/kg/day.

In another aspect of the present invention, the composition provides a composition for topical administration.

In another aspect of the present invention, there is provided a composition, wherein the composition is an ophthalmic formulation.

The compound isolated from Centella asiatica extract of the present invention, asiaticoside or asiatic acid, increases glucose metabolism to effectively proliferate retinal cells and protects retinal cells from oxidative damage. Therefore, asiaticoside and asiatic acid, which are compounds isolated from Centella asiatica extract, can improve, prevent or treat eye diseases caused by retinal cell damage, that is, glaucoma and macular degeneration.

The effects of the present invention are not limited to the above-mentioned effects. It should be understood that the effects of the present invention include all effects that can be inferred from the following description.

1 shows the chromatography results of Centella asiatica extract.
Figure 2 shows the retinal cell proliferation effect of the compounds of the present invention, asiaticoside (2a) and asiatic acid (2b).
Figure 3 shows the protective effect on oxidative stress by treatment with the compound of the present invention in retinal pigment epithelial cells in terms of cell viability. Figure 3a relates to asiaticoside, and Figure 3b relates to asiatic acid.
Figure 4 shows the glucose metabolism effect of the compound of the present invention in the retinal pigment epithelial cells.

Unless otherwise specified, all numbers, values, and/or expressions expressing ingredients, reaction conditions, and amounts of ingredients used herein refer to a variety of measures that may occur in obtaining such values, among others, in which such numbers are inherently different. Since they are approximations reflecting uncertainty, it should be understood as being modified by the term "about" in all cases. Also, where the disclosure discloses numerical ranges, such ranges are continuous and inclusive of all values from the minimum to the maximum inclusive of the range, unless otherwise indicated. Furthermore, when such ranges refer to integers, all integers inclusive from the minimum to the maximum inclusive are included, unless otherwise indicated.

In this specification, when a range is described for a variable, the variable will be understood to include all values within the stated range including the stated endpoints of the range. For example, a range of “5 to 10” includes the values of 5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc. It will be understood to include any value between integers that are appropriate for the scope of the recited range, such as 5.5, 6.5, 7.5, 5.5 to 8.5 and 6.5 to 9, and the like. Also for example, a range of "10% to 30%" includes values such as 10%, 11%, 12%, 13%, and all integers up to and including 30%, as well as 10% to 15%, 12% to It will be understood to include any subrange, such as 18%, 20% to 30%, etc., as well as any value between reasonable integers within the scope of the recited range, such as 10.5%, 15.5%, 25.5%, and the like.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for preventing, improving or treating retinal diseases, comprising a compound isolated from Centella asiatica extract as an active ingredient. Specifically, the composition induces proliferation of retinal cells by increasing glucose metabolism, and protects cells from oxidative stress to prevent and improve or treat retinal diseases such as glaucoma and macular degeneration for eye health. there is.

Centella asiatica is a perennial creeping herb belonging to the Umbrella family, and grows wild in hot and humid areas. , 2006). The main known components of Centella asiatica are triterpenic acid sugar esters, and asiaticoside, madecassoside, brahmoside, brahminoside, etc. have been reported, and it is known that asiatic acid, madecassic acid, thankunic acid, isothankunic acid, etc. are produced during hydrolysis. As a major component of Centella asiatica, asiaticoside and madecassoside, which are pentacyclic triterpene glycosides belonging to the α-amyrin-ursolic acid group, have been used for the treatment of skin wounds and chronic ulcers for a long time. It has been reported to have a therapeutic effect on psychiatric diseases, tuberculosis, venous disease, and dementia. Due to these effects, Centella asiatica is used for various purposes such as food, skin treatment, wound treatment, memory enhancer, and tonic. However, it has not been stated that asiaticoside and asiatic acid, the main components of Centella asiatica extract, are effective in preventing, improving or treating retinal diseases such as glaucoma and macular degeneration.

Accordingly, the present invention provides a composition for preventing, improving or treating retinal diseases, comprising an asiaticoside compound represented by the following Chemical Formula 1, or a pharmaceutically acceptable salt thereof, as an active ingredient.

[Formula 1]

The asiaticoside of the present invention is a triterpene saponin compound, whose chemical names are 2α, 3β, 23-trihydroxyurs-12-en-28-oic acid α-L-rhamnopyranosyl-(1-4)o-β-D-gluco- A pyranosyl ester can be isolated from a naturally occurring, commercially available one (eg, SIGMA-ALDRICH), or synthesized by a method known in the art.

In addition, the present invention provides a composition for preventing, improving or treating retinal diseases, comprising an asiatic acid compound represented by the following Chemical Formula 2, or a pharmaceutically acceptable salt thereof, as an active ingredient.

[Formula 2]

The chemical name of the asiatic acid of the present invention is 2α, 3β, 23-trihydroxyurs-12-en-28-oic acid, which can be isolated from natural ones, and can be purchased commercially (eg, SIGMA- ALDRICH), can be synthesized by methods known in the art.

The Centella asiatica extract may be extracted by a conventional extraction method, and may be an extract having a powdered form by drying the extract under reduced pressure and freeze-drying.

In one aspect or another aspect of the present invention, the Centella asiatica extract provides a composition, which is an extract extracted with water, a lower alcohol of C1-6, or a mixed solvent thereof.

For the composition for preventing and improving or treating glaucoma and macular degeneration having the above composition of the present invention, the extraction process may be repeated as necessary, and an extract extracted 2 to 5 times may be used. In addition, a dry powdery extract obtained by freeze-drying the extract can be prepared and used in the composition for preventing and improving or treating glaucoma and macular degeneration.

The content of the alcohol in the extraction solvent is good in terms of extraction efficiency to maintain a concentration range of 0 to 95% by weight of alcohol, preferably 30 to 95% by weight, and more preferably 50% by weight. Such alcohol can be applied as long as it is generally used in the art, and preferably, an alcohol having 1 to 5 carbon atoms, the alcohol may be one or more selected from methanol, ethanol, isopropanol, propanol and butanol. More preferably, the alcohol may be ethanol, alcohol, or the like.

In one aspect or another aspect of the present invention, the Centella asiatica extract is 30% to 90% of an ethanol aqueous solution extract, it provides a composition.

The extraction method is a method commonly known in the art, for example, a method using an extraction device such as supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction or ultrasonic extraction, or an adsorption resin including XAD and HP-20. How to use, etc. can be used.

In addition, the extract is concentrated under reduced pressure using a vacuum rotary evaporator or the like to obtain an extract. In addition, the obtained extract can also be dried under reduced pressure, vacuum drying, boiling drying, spray drying, room temperature drying, freeze drying, etc. as needed. In particular, when the method of freeze-drying is applied, there is an advantage that the loss of volatile organic substances in the extract can be reduced.

In one aspect of the present invention, the Centella asiatica extract provides a composition, which is a 40% to 60% ethanol aqueous solution extract.

In one aspect of the present invention, the composition provides a composition, characterized in that it proliferates optic nerve cells and retinal cells or protects cells from oxidative stress in eye health.

In one aspect or another aspect of the present invention, the Centella asiatica extract provides a composition, which is administered in a dosage of 0.01 mg to 10 g/kg/day.

Various aspects of the present invention are described below.

One aspect of the present invention provides an asiaticoside compound represented by Formula 1 below, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof or a solvate thereof; and an asiatic acid compound represented by Formula 2 below, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof or a solvate thereof; It provides a composition for improving, preventing or treating retinal cell damage diseases caused by oxidative stress, comprising any one or more of them as an active ingredient.

[Formula 1]

[Formula 2]

In one aspect of the present invention, the active ingredient is an asiaticoside compound represented by Formula 1, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof or a solvate thereof. It provides a composition for improving, preventing or treating retinal cell damage diseases.

In one aspect of the present invention, the active ingredient is an asiaticoside compound represented by Formula 1, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof or a solvate thereof; And an asiatic acid compound represented by Formula 2 below, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof or a solvate thereof. A composition for improving, preventing or treating is provided.

As used herein, "isomers" are in particular optical isomers (e.g., essentially pure enantiomers, essentially pure diastereomers or mixtures thereof), as well as conformational isomers ( includes conformation isomers (i.e. isomers that differ only in the angle of one or more chemical bonds), position isomers (especially tautomers) or geometric isomers (e.g. cis-trans isomers) do.

As used herein, "essentially pure", for example, when used in reference to enantiomers or diasteres, contains at least about 90%, preferably about 95%, of a specific compound exemplified by enantiomers or diisomers. or more, more preferably about 97% or more, or about 98% or more, even more preferably about 99% or more, even more preferably about 99.5% or more (w/w).

As used herein, "pharmaceutically acceptable" is approved by the government or equivalent regulatory body for use in animals, more specifically humans, by avoiding significant toxic effects when used in conventional medical dosages. means that it is available for, or has been approved for, or recognized as being listed in a pharmacopoeia or other general pharmacopoeia.

As used herein, "pharmaceutically acceptable salt" refers to a salt according to an aspect of the present invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. The salts are formed by (1) inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid , 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2,2,2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, acid addition salts formed with organic acids such as tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid; or (2) salts formed when an acidic proton present in the parent compound is substituted.

As used herein, the term “hydrate” refers to a compound to which water is bonded, and is a broad concept including inclusion compounds that do not have a chemical bonding force between water and the compound.

As used herein, the term “solvate” refers to a higher-order compound formed between a molecule or ion of a solute and a molecule or ion of a solvent.

The compounds of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates and alkanes. It is obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, ioda. Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate , sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Toxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate or mandelate.

In one aspect of the present invention, the compound represented by Formula 1 and the compound represented by Formula 2 are each independently isolated or synthesized from Centella asiatica extract, improvement of retinal cell damage caused by oxidative stress , to provide a composition for prevention or treatment.

In one aspect of the present invention, the compound represented by Formula 1 and the compound represented by Formula 2 are each independently isolated from Centella asiatica extract, improvement, prevention or treatment of retinal cell damage caused by oxidative stress A composition is provided.

In one aspect of the present invention, the composition provides a composition, characterized in that the survival rate of retinal cells is improved by increasing the glucose availability of the retinal cells.

In one aspect of the present invention, the disease is a retinal disease, macular degeneration or glaucoma, provides a composition.

In one aspect of the present invention, the prophylactic or therapeutic composition is a pharmaceutical composition, it provides a composition. In one embodiment, the composition provides a pharmaceutical composition, administered in a dosage of 0.01 mg/kg/day to 10 g/kg/day. In one embodiment, the composition provides a pharmaceutical composition, which is a formulation for topical administration. In one embodiment, the composition provides an ophthalmic formulation, a pharmaceutical composition.

Another aspect of the present invention provides a pharmaceutical composition for preventing, improving or treating retinal cell damage caused by oxidative stress, comprising a compound isolated from Centella asiatica extract as an active ingredient.

In another aspect of the present invention, the compound isolated from the Centella asiatica extract provides a pharmaceutical composition that improves the viability of retinal cells by increasing sugar availability.

In another aspect of the present invention, the compound isolated from the Centella asiatica extract provides a pharmaceutical composition that prevents the death of retinal cells from oxidative stress.

In another aspect of the present invention, the disease is macular degeneration or glaucoma, to provide a pharmaceutical composition.

When the pharmaceutical composition is used clinically, it is formulated with a carrier conventional in the pharmaceutical field to prepare conventional formulations in the pharmaceutical field, for example, tablets, capsules, powders, granules, pills, liquids and suspensions. formulations for oral administration; Injection preparations in the form of solutions or suspensions for injection, or ready-to-use dry powder for injection that can be prepared with distilled water for injection at the time of injection; Or it can be formulated into various preparations such as ointments. Pharmaceutical preparations prepared using conventional carriers can be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally or topically. Accordingly, the pharmaceutical composition of the present invention may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceuticals.

Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate , cellulose, methyl cellulose, hydroxymethyl cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, povidone, crospovidone, croscarmellose sodium, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, noi At least one selected from sirin, colloidal silicon dioxide, lactose, talc, magnesium stearate, colloidal magnesium stearyl, and mineral oil may be included.

In the case of formulation, it is prepared using diluents or excipients, such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, troches, rosins, capsules, and the like, and these solid preparations include at least one excipient in the composition of the present invention, for example, lactose, saccharose, sorbitol. , mannitol, starch, amylopectin, cellulose, calcium carbonate, gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral administration include suspensions, solutions, emulsions, elixirs, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol gelatin and the like can be used. For parenteral administration, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection may be common.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the patient's age, weight, disease severity, drug form, administration route and period, but may be appropriately selected by those skilled in the art. However, for a desirable effect, the pharmaceutical composition of the present invention may be administered at 0.01 mg/kg to 10 g/kg per day, preferably at 1 mg/kg to 1 g/kg. Administration may be administered several times a day at regular time intervals according to the judgment of a doctor or pharmacist, preferably 1 to 6 divided doses.

In one embodiment, the composition is a natural compound that has been ingested and taken on a daily basis and can be safely used for preventing, improving, or treating glaucoma and macular degeneration.

In one or another aspect of the present invention, there is provided a composition wherein the compound isolated from the Centella asiatica extract is administered at a dosage of 0.01 mg to 10 g/kg/day.

In another aspect of the present invention, the composition provides a composition for topical administration.

In another aspect of the present invention, there is provided a composition, wherein the composition is an ophthalmic formulation.

In one aspect or another aspect of the present invention, the compound isolated from the Centella asiatica extract is included in the composition at a concentration of 5 to 500 μg/ml based on the total weight of the composition, and provides a composition.

The composition effectively proliferates retinal cells and has an effect of protecting the cells from oxidative stress. In addition, the composition can be safely used for preventing, improving or treating glaucoma and macular degeneration as a natural compound that has been ingested and taken on a daily basis.

In one aspect of the present invention, the composition for preventing or improving is a health functional food composition, it provides a composition.

One aspect of the present invention provides a food composition for protecting retinal cells for protecting retinal cells from oxidative stress, comprising a compound isolated from Centella asiatica extract as an active ingredient.

The health functional food composition according to the present invention contains asiaticoside or asiatic acid as an active ingredient, a compound isolated from Centella asiatica extract, and can be ingested for the purpose of improving eyesight through inhibition of glaucoma and macular degeneration, which are retinal diseases. The active ingredient may be ingested as food prepared in tablets, capsules, powders, granules, pills, liquids, suspensions, or the like, or added to general food. Since the health functional food uses food as a raw material unlike general drugs, there is an advantage in that there are no side effects that may occur when taking the drug for a long time. The content of the active ingredient may be appropriately determined depending on the purpose of its use (for prevention or improvement). In general, the active ingredient in the health functional food composition may be included in 0.1 to 90% by weight. However, in the case of long-term ingestion for health and hygiene purposes or for health control, the above amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There is no particular limitation on the type of the food. Examples of foods to which the active ingredient can be added include drinks, meat, sausage, bread, biscuits, rice cakes, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, and various soups. , beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products, and includes all health functional foods in the ordinary sense. The properties of the food are also not particularly limited, and may be any of a solid form, a semi-solid form, a gel form, a liquid form, a powder form, and the like.

It can be prepared as a beverage by using the health functional food composition of the present invention. As a component included in the beverage, there is no particular limitation in the selection of other components other than the active component, and it may contain various flavoring agents or natural carbohydrates as additional components as in a conventional beverage. 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 polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition, the health functional food composition of the present invention, in addition to the active ingredient, various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and Salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like may be contained as additives. In addition, it may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination. Although the proportion of these additives is not so important, it is generally selected in the range of 0.1 to 20% by weight in the health functional food composition of the present invention.

In one aspect of the present invention, the compound isolated from the Centella asiatica extract provides a food composition that improves the viability of retinal cells through increased sugar availability.

In one aspect of the present invention, the compound isolated from the Centella asiatica extract provides a food composition that prevents the death of retinal cells from oxidative stress.

In one aspect of the present invention, the food composition provides a food composition for preventing or improving retinal cell damage caused by oxidative stress.

Hereinafter, the present invention will be described in more detail through specific examples. The following examples are merely examples to help the understanding of the present invention, and the scope of the present invention is not limited thereto.

[Examples and Comparative Examples]

Preparation Example: Preparation of Centella asiatica extract

Centella asiatica was purchased from a farmhouse in Hapcheon, Gyeongnam, and dried in the shade, and then 1 kg of crushed centella asiatica was extracted with 50% alcohol at 80° C. for 5 hours, and then concentrated and freeze-dried to 135 g of Gujeolcho extract (yield 13.5%). ) was obtained.

[Experimental example]

Experimental Example 1: Analysis of asiaticoside and asiatic acid content of Centella asiatica extract

Centella asiatica extract obtained in Preparation Example was analyzed using high performance liquid chromatography (HPLC).

For the analysis of Centella asiatica extract, HPLC (LC-20AD, Shimadzu Corp., Japan) was used, and after diluting the Centella asiatica extract of the preparation example to a concentration of 10 mg/mL, it was filtered with a 0.45 μm syringe filter to add 10 μL to HPLC. It was analyzed by injection. As the analytical column, Skypak C18 (250X4 mm) was used, and the mobile phase A used for analysis was distilled water and the mobile phase B used was an acetonitrile solution. Mobile solvent conditions were initially 80% for mobile phase A and 20% for mobile phase B, and at 40 minutes, 20% for mobile phase A and 80% for mobile phase B. The mobile phase flow rate was 1.0 mL/min, and analysis was performed at a UV wavelength of 206 nm.

The chromatography results of each component are shown in FIG. 1 .

As shown in FIG. 1, asiaticoside and asiatic acid compound were detected in the 50% ethanol extract of Centella asiatica, and the content was 77.4±19.92 and 5.2±0.13 mg/g, respectively.

Experimental Example 2: Measurement of retinal cell proliferation effect

The protective and proliferative effects of retinal cells were measured using asiaticoside and asiatic acid isolated from the Centella asiatica extract.

ARPE-19 retinal pigment epithelial cells were cultured in DMEM/F12 medium containing 10% serum and antibiotics, supplied with 5% by volume of carbon dioxide and maintained at 37°C. ARPE-19 cells were aliquoted in a 96-well plate at a concentration of 5 × 10 ^{3 cells/well, and cultured for 24 hours.} In order to measure the proliferative effect of asiaticoside and asiatic acid on retinal pigment epithelial cells, after exchange with DMEM/F12 medium containing 10% charcoaled FBS, each compound was treated at 5, 10, and 20 μg/mL. 20 μM of lutein was further treated as a control. After 48 hours, each cell growth rate was measured at absorbance at 550 nm using MTT reagent, and the absorbance of each test solution was measured based on the control to indicate the cell proliferation rate (%).

Retinal pigment epithelial cell proliferation rates of asiaticoside and asiatic acid are shown in FIG. 2 .

As shown in FIG. 2 , asiaticoside and asiatic acid showed very good proliferation efficacy at a concentration of 20 μg/mL, and compared with the control group, asiaticoside increased by 12.2% and asiatic acid by 40.7%.

Experimental Example 3: Measurement of the protective effect of oxidative damage in retinal cells

ARPE-19 retinal pigment epithelial cells were cultured in the same manner as in Experimental Example 2, and _{after inducing oxidative damage to cells using cobalt chloride (CoCl 2} ), the protective effect of asiaticoside and asiatic acid was measured.

The retinal pigment epithelial cell line ARPE-19 was aliquoted into a 96-well plate at a concentration ^{of 3Х10 4} _{cells/well, and cultured at 37° C., 5% CO 2 in a} cell incubator. Cells were pretreated with asiaticoside and asiatic acid at a concentration of 2.5, 5, and 10 μg/mL, respectively, for 2 hours, and after 2 hours, 500 μM cobalt chloride was simultaneously treated for 24 hours. Thereafter, each cell protective effect was measured at absorbance at 550 nm using MTT reagent, and the absorbance of each test solution was measured based on the control to indicate the cell protective effect (%).

The protective effect of asiaticoside and asiatic acid against oxidative damage is shown in FIG. 3 .

As shown in FIG. 3 , asiaticoside and asiatic acid showed superior protective effects against oxidative damage compared to the control group, and at a concentration of 10 μg/mL, asiaticoside was 21.5% higher than that of the cobalt chloride-treated control group, respectively. Tic acid increased the cell viability by 27.6%.

Experimental Example 4: Measurement of Efficacy of Glucose Metabolism in Retinal Epithelial Cells

A 2-NBDG uptake experiment was conducted to study the glucose uptake effect of the asiaticoside and asiatic acid in retinal pigment epithelial cells.

ARPE-19, a retinal pigment epithelial cell line, was used as the test cell, and the medium, medium composition, and experimental conditions were the same as in Experimental Example 2 above.

ARPE-19 cells were aliquoted in a 96-well plate at a concentration of 1 × 10 ^{4 cells/well, and cultured for 24 hours.} To measure the glucose metabolism efficacy, asiaticoside and asiatic acid were treated at 5, 10, and 20 μg/mL, incubated for 24 hours, and then serum-free DMEM/F12 medium and 100 μM 2-NBDG were treated for 30 minutes. did. After washing twice with cold phosphate buffered saline (PBS), the amount of glucose uptake was measured with a fluorescence photometer (excitation 485 nm; emission 525 nm).

The glucose uptake effect of asiaticoside and asiatic acid is shown in FIG. 4 .

As shown in FIG. 4, both asiaticoside and asiatic acid showed excellent glucose metabolism efficacy in retinal cells, and in particular, showed higher glucose metabolism efficacy than insulin, a positive control, so this result shows that asiaticoside and asiatic acid It can be said to be the basis for the retinal cell proliferation efficacy of

Accordingly, the present invention can provide a composition for preventing, improving, or treating retinal diseases such as glaucoma and macular degeneration for eye health, including asiaticoside or asiatic acid, which is a compound isolated from Centella asiatica extract.

[Formulation example]

On the other hand, the pharmaceutical composition comprising the active ingredient according to the present invention can be prepared in various forms depending on the purpose. The following formulations 1 to 4 exemplify the pharmaceutical preparation method containing the active ingredient according to the present invention, but the present invention is not limited thereto.

Formulation 1: Tablet (Direct Pressurization)

After 5.0 mg of the active ingredient was sieved, lactose 14.1 mg, crospovidone USNF 0.8 mg, and magnesium stearate 0.1 mg were mixed and pressurized to make tablets.

Formulation 2: Tablet (wet granulation)

After sieving 5.0 mg of the active ingredient, 16.0 mg of lactose and 4.0 mg of starch were mixed. After dissolving 0.3 mg of polysorbate 80 in pure water, an appropriate amount of this solution was added, followed by atomization. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressurized to make tablets.

Formulation 3: Powders and Capsules

After sieving 5.0 mg of the active ingredient, it was mixed with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. Mix the mixture into a solid No. Filled in 5 gelatin capsules.

Formulation 4: Injection

An injection was prepared by containing 100 mg as an active ingredient, and also containing 180 mg of mannitol, _{26 mg of Na 2} HPO ₄ 12H ₂ O, and 2974 mg of distilled water.

In addition, the health food composition comprising the active ingredient according to the present invention can be prepared in various forms depending on the purpose. The following formulations 5 to 9 exemplify the manufacturing method of a health food containing the active ingredient according to the present invention, but the present invention is not limited thereto.

Formulation 5. Granular Health Food

Active ingredient 1000 mg, vitamin A acetate 70 µg, vitamin E 1.0 mg, vitamin 0.13 mg, vitamin B ₂ 0.15 mg, vitamin B ₆ 0.5 mg, vitamin B ₁₂ 0.2 µg, vitamin C 10 mg, biotin 10 µg, nicotinic acid amide 1.7 mg, folic acid 50 μg, calcium pantothenate 0.5 mg, 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 and magnesium chloride 24.8 mg were mixed, and then granular health food was prepared according to a conventional method.

Formulation 6. Health drink

Active ingredient 1000 mg, citric acid 1000 mg, oligosaccharide 100 g, plum concentrate 2 g, and taurine 1 g were mixed, and purified water was added thereto to adjust the total volume to 900 ml. After stirring and heating at 85° C. for about 1 hour, the resulting solution was filtered and obtained in a sterilized 2L container, sealed and sterilized to prepare a health drink.

Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demanding class, demanding country, and use.

Formulation 7. Flour Foods

0.5 to 5 g of the active ingredient was added to 100 g of wheat flour, and the mixture was used to prepare breads, cakes, cookies, crackers and noodles to prepare health-promoting foods.

Formulation 8. Dairy Products

5 to 10 g of the active ingredient was added to 100 g of milk, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation 9. Seonshik

30 g of brown rice, 20 g of barley, 10 g of glutinous rice, and 15 g of barley radish were prefabricated by a known method, dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder. 7 g of black beans, 7 g of black sesame seeds, and 7 g of perilla sesame seeds were steamed and dried by a known method, and then dried to prepare powder having a particle size of 60 mesh using a grinder. A wire was prepared by mixing 3 g of the active ingredient of the present invention with the grains and seeds prepared above.

As mentioned above, although embodiments of the present invention have been described with reference to the accompanying drawings, those of ordinary skill in the art to which the present invention pertains can practice the present invention in other specific forms without changing its technical spirit or essential features. You will understand that there is Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (12)

An asiaticoside compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, a hydrate or a solvate thereof, as an active ingredient, and improvement in the survival rate of retinal pigment epithelial cells or oxidative oxidation of retinal pigment epithelial cells A pharmaceutical composition for improving, preventing or treating macular degeneration, improving, preventing or treating macular degeneration through stress suppression.
[Formula 1]

An asiaticoside compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, a hydrate or a solvate thereof, as an active ingredient, and improvement in the survival rate of retinal pigment epithelial cells or oxidative oxidation of retinal pigment epithelial cells A health functional food composition for improving or preventing macular degeneration, improving or preventing macular degeneration through stress suppression.
[Formula 1]

3. The method of claim 1 or 2,
The composition further comprises an asiatic acid compound represented by Formula 2 below, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.
[Formula 2]

.
3. The method of claim 1 or 2,
The compound represented by Formula 1 is isolated from or synthesized from Centella asiatica extract, the composition.
4. The method of claim 3,
The compound represented by Formula 2 is a composition that is isolated or synthesized from Centella asiatica extract.
3. The method of claim 1 or 2,
The composition is characterized in that it improves the survival rate of the retinal pigment epithelial cells by increasing the glucose availability of the retinal pigment epithelial cells, the composition.
delete delete delete The method of claim 1,
The composition is administered in a dosage of 0.01 mg / kg / day to 10 g / kg / day, the composition.
The method of claim 1,
The composition is a formulation for topical administration.
The method of claim 1,
The composition is an ophthalmic formulation.

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