KR20200107744A - 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 and alleviating or treating retinal diseases such as glaucoma and macular degeneration, comprising a compound isolated from a Centella asiatica extract as an active ingredient and, more specifically, to a composition which increases the efficiency of glucose metabolism of cells, thereby increasing the survival rate of retinal cells, and protects retinal cells from oxidative damage, thereby being usefully used as a composition for preventing and alleviating or treating retinal diseases such as glaucoma and macular degeneration for eye health.

Description

Composition for prevention or treatment of retinal diseases comprising compounds isolated from Centella asiatica extract, comprising a compound isolated from centella asiatica extract as an active ingredient

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 caused by damage to the optic nerve due to an increase in intraocular pressure or abnormal blood circulation of the optic nerve. It is characterized by temporary or permanent visual field defects and decreased vision. If not treated, it can lead to permanent blindness. The general cause of the outbreak is increased intraocular pressure due to circulatory disorders in the eyeball, and when the age group of 40s or older, the waterproof outlet becomes narrower and the amount of waterproofing produced increases, the intraocular pressure rises, which causes pressure and damage to the optic nerve. . In addition, although intraocular pressure is normal, glaucoma develops in some cases, especially in Korea and Japan, there are more patients with glaucoma with normal intraocular pressure (about 70-80% of glaucoma patients), and the cause is known as an optic nerve blood circulation disorder. Practically, it is known that there are many cases of peripheral blood circulation disorders caused by metabolic diseases in patients within the range of normal intraocular pressure. Ultimately, glaucoma leads to the death of cells such as retinal node cells (Almasieh et al., 2012), and substances that can prevent and treat such cell death can be used as foods and therapeutics to inhibit the onset and progression of glaucoma. .

The macula is the part that receives light most clearly and accurately because the eye cells are concentrated in the retina of the eye. It plays a very important role in vision. The macular degeneration occurs due to various causes and causes vision disorders. (macular degeneration). Macular degeneration is a disease caused by gradual degeneration of the retinal epithelial cells, the retina and the capillary layer of the choroid. In the early stages, macular degeneration is an extracellular deposit called drusen between the Bruch's membrane and the retinal epithelial cells. It is characterized by the observation of abnormal retinal epithelial cells formed. Also, in the later stages, it is largely divided into exudative (wet) macular degeneration and atrophic (dry) macular degeneration.Since the optic cells in the macula are gradually destroyed in both forms of macular degeneration, the function of the macular deteriorates over time and central vision decreases. Decrease. Macular degeneration is known to be the main cause of oxidative stress and inflammation. In particular, when by-products after intracellular metabolism are not excreted in retinal pigment epithelial cells but gradually accumulate, it undergoes an oxidative stress process and 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 the death of optic nerve cells or retinal nerve cells can be said to be the main causes, and these two diseases have recently increased the prevalence of metabolic diseases, as well as 30-50 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 preventing oxidative damage to the eyes, has been actively developed. In particular, energy improvement agents using anthocyanoside derived from wild blueberry as a main component have been developed. Herbal medicines have been used for the purpose of maintaining vision and improving visual acuity from ancient times, but there are no studies on how to prevent or treat glaucoma related to retinal disease and macular degeneration of the retina using major components of centella asiatica.

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

Accordingly, the present inventors recognize the importance of prevention and treatment for retinal diseases such as glaucoma and macular degeneration, and use a variety of natural materials and main ingredients to provide natural resources that can protect the retinal pigment epithelial cell proliferation efficacy and oxidative damage. Screened. As a result, Centella asiatica extract showed excellent proliferative efficacy in optic nerve cells and retinal cells, inhibited oxidative damage caused by A2E in retinal cells, and significantly increased sugar availability in retinal cells. In particular, asiaticoside and asiatic acid, the compounds isolated from centella asiatica extract, had the survival rate of retinal pigment epithelial cells and cytoprotective effects from oxidative stress. Increased. Since this can be inferred that the recovery effect and the protective effect according to cell damage are excellent, the compound isolated from the centella asiatica extract of the present invention is highly likely to be commercialized as a composition for the prevention and 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 preventing and improving 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 object of the present invention will become more apparent from the following description, and will be realized by the means described in the claims and combinations thereof.

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

In one aspect of the present invention, asiaticoside or asiatic acid as 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, a compound isolated from the centella asiatica extract, provides a food composition that prevents the death of retinal cells from oxidative stress of 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 diseases caused by oxidative stress, comprising asiaticoside or asiatic acid or a pharmaceutically acceptable salt, which is a compound isolated from centella asiatica extract, as an active ingredient. It provides a pharmaceutical composition for.

In another aspect of the present invention, asiaticoside or asiatic acid, a compound isolated from the centella asiatica extract, provides a pharmaceutical composition for improving the survival rate of retinal cells by increasing sugar availability.

In another aspect of the present invention, asiaticoside or asiatic acid, a compound isolated from the centella asiatica extract, provides a pharmaceutical composition for preventing the death of retinal cells from oxidative stress.

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

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

In one aspect or another aspect of the present invention, 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, providing a composition.

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

In another aspect of the present invention, the composition provides a composition, which is an ophthalmic formulation.

Asiaticoside or asiatic acid, a compound isolated from centella asiatica extract of the present invention, increases sugar metabolism, effectively proliferates 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, that is, glaucoma and macular degeneration caused by retinal cell damage.

The effects of the present invention are not limited to the effects mentioned above. 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 chromatographic 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).
3 shows the protective effect against oxidative stress by the treatment of the compound of the present invention in retinal pigment epithelial cells in terms of cell survival, FIG. 3A is asiaticoside, and FIG. 3b is for asiatic acid.
Figure 4 shows the glucose metabolism effect by the treatment of the compound of the present invention in retinal pigment epithelial cells.

Unless otherwise specified, all numbers, values, and/or expressions expressing ingredients, reaction conditions, and content of ingredients used herein are the variety of measurements that occur in obtaining such values, among other things, in which these numbers are essentially different. Since they are approximations that reflect uncertainty, they should be understood as being modified in all cases by the term "about". In addition, when numerical ranges are disclosed herein, these ranges are continuous and, unless otherwise indicated, include all values from the minimum value of this range to the maximum value including the maximum value. Furthermore, when this range refers to an integer, all integers from the minimum value to the maximum value including the maximum value are included, unless otherwise indicated.

In the present specification, when a range is described for a variable, it will be understood that the variable includes all values within the stated range, including the stated endpoints of the range. For example, a range of "5 to 10" includes 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. Inclusive, and it will be understood to include any values between integers that are reasonable in the scope of the stated 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, the range of "10% to 30%" is 10% to 15%, 12% to 10%, 11%, 12%, 13%, etc., as well as all integers including up to 30%. It will be understood to include any subranges such as 18%, 20% to 30%, and the like, and include any values between reasonable integers within the scope of the stated 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 increases sugar metabolism, induces proliferation of retinal cells, and protects cells from oxidative stress and can be usefully used as a composition for preventing and improving retinal diseases such as glaucoma and macular degeneration for eye health. have.

Centella asiatica is a perennial creeping herb belonging to the umbel family, and grows naturally in hot and humid areas. , 2006). Asiaticoside, madecassoside, brahmoside, brahminoside, etc. are reported as triterpenic acid sugar esters, and asiatic acid, madecassic acid, thankunic acid, and isothankunic acid are known to be produced during hydrolysis. Asiaticoside and madecassoside, pentacyclic triterpene glycosides belonging to the α-amyrin-ursolic acid group as major components of centella, have long been used in the treatment of skin wounds and chronic ulcers. , Mental disease, tuberculosis, venous disease, and dementia are reported to have therapeutic effects. Due to these effects, Centella asiatica is used for various purposes such as edible, skin treatment, wound treatment, memory enhancer, and tonic. In Korea, Centella asiatica is used in the fields of pharmaceuticals, cosmetics, and functional foods. Also, asiaticoside and asiatic acid, the main ingredients of centella asiatica extract, have not been disclosed to be 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 as an active ingredient an asiaticoside compound represented by the following Formula 1, or a pharmaceutically acceptable salt thereof.

[Formula 1]

The asiaticoside of the present invention is a triterpene saponin compound, and the chemical names are 2α, 3β, 23-trihydroxyurs-12-en-28-oic acid α-L-rhamnopyranosyl-(1-4)o-β -D-gluco- The pyranosyl ester can be used to separate what exists in nature, and commercially available ones can be purchased (for example, SIGMA-ALDRICH), or can be synthesized by methods 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 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 those that exist in nature, and commercially available ones can be purchased (e.g., SIGMA- ALDRICH), can be synthesized by a method known in the art.

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

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

In the composition for preventing, improving or treating glaucoma and macular degeneration having the above composition of the present invention, the extraction process may be repeated as needed, and an extract extracted repeatedly 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 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. These alcohols may be applied as long as they are generally used in the art, preferably alcohols having 1 to 5 carbon atoms, and the alcohol may be one or more selected from methanol, ethanol, isopropanol, propanol, and butanol. More preferably, ethanol, alcohol, etc. may be used as the alcohol.

In one aspect or another aspect of the present invention, the centella asiatica extract is a 30% to 90% ethanol aqueous solution extract, providing 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. A method of use or the like 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 may be dried under reduced pressure, vacuum drying, boiling drying, spray drying, room temperature drying, or freeze drying, if necessary. In particular, there is an advantage in that the loss of volatile organic substances in the extract can be reduced when the freeze-drying method is applied.

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

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 is administered at a dosage of 0.01mg to 10g/kg/day, providing a composition.

Hereinafter, various aspects of the present invention will be described.

An aspect of the present invention is an asiaticoside compound represented by the following 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; It provides a composition for improving, preventing or treating diseases of retinal cell damage 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, due to oxidative stress. 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 Chemical Formula 1, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate or a solvate thereof; And an asiatic acid compound represented by the following formula (2), an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof. It provides a composition for improvement, prevention or treatment.

"Isomers" in the present specification are in particular optical isomers (e.g., essentially pure enantiomers, essentially pure diastereomers or mixtures thereof), as well as conformational isomers ( 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.

In the present specification, the term "essentially pure", for example, when used in connection with an enantiomer or diastereomer, contains about 90% or more, preferably about 95%, of a specific compound exemplified by an enantiomer or diastereomer. Or more, more preferably at least about 97% or at least about 98%, even more preferably at least about 99%, and even more preferably at least about 99.5% (w/w).

"Pharmacologically acceptable" as used herein means government or equivalent regulatory approval that it can be used in animals, more specifically in humans, by avoiding significant toxic effects when used in conventional medicinal dosages. It means to be able to receive or be approved, or to be recognized as listed in the pharmacopoeia or as described in other general pharmacopoeia.

In the present specification, "pharmaceutically acceptable salt" refers to a salt according to an aspect of the present invention that is pharmaceutically acceptable and has a desirable pharmacological activity of a parent compound. The salt is (1) formed of an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or 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 of organic acids such as tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid; Or (2) a salt formed when the acidic proton present in the parent compound is substituted.

In the present specification, "hydrate" refers to a compound to which water is bound, and is a broad concept including a compound containing no chemical bonding force between water and the compound.

In the present specification, "solvate" refers to a high-order compound formed between molecules or ions of a solute and molecules or ions of a solvent.

The compounds of the present invention can be used in the form of a pharmaceutically acceptable salt, and 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, hydroxy alkanoates 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, butin-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, me Oxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hydroxybutyrate, glycolate, Maleate, 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, improving retinal cell damage diseases caused by oxidative stress It provides a composition for preventing or treating.

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, improving, preventing or treating retinal cell damage diseases caused by oxidative stress It provides a composition for use.

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

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

In one aspect of the present invention, the composition for preventing or treating is a pharmaceutical composition, providing a composition. In one embodiment, the composition is administered at a dosage of 0.01mg / kg / day to 10g / kg / day, provides a pharmaceutical composition. 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 diseases 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 for improving the survival rate 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 for preventing the death of retinal cells from oxidative stress.

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

When the pharmaceutical composition is used clinically, it is mixed with a conventional carrier in the pharmaceutical field, such as tablets, capsules, powders, granules, pills, liquids, and suspensions. Formulations for oral administration; Injectable preparations in the form of a solution or suspension for injection, or a ready-to-use dry powder for injection that can be prepared and used with distilled water for injection upon injection; Alternatively, it may be formulated in various formulations such as ointments. Pharmaceutical preparations prepared by using a conventional carrier can be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally or topically. Accordingly, the pharmaceutical composition of the present invention may further include an appropriate carrier, excipient, and diluent commonly used in the manufacture of a medicament.

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, gum acacia, 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 Syrin, colloidal silicon dioxide, lactose, talc, magnesium stearate, colloidal magnesium stearyl, and one or more selected from mineral oil may be included.

In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, troches, rosin papers, capsules, etc., and these solid preparations are at least one excipient in the composition of the present invention, such as 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, solvents, emulsions, elixirs, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as humectants, sweeteners, fragrances, preservatives, etc. May be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol gelatin, and the like may be used. Parenteral administration may generally be a subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the patient's age, weight, degree of disease, drug form, administration route and duration, 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 1 mg/kg to 1 g/kg. Administration may be divided into several times a day, preferably once to six times, at regular time intervals according to the judgment of a doctor or pharmacist.

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 aspect or another aspect of the present invention, the compound isolated from the centella asiatica extract is administered at a dosage of 0.01mg to 10g/kg/day, providing a composition.

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

In another aspect of the present invention, the composition provides a composition, which is an ophthalmic formulation.

In one aspect or another aspect of the present invention, the compound isolated from the centella asiatica extract is contained in the composition at a concentration of 5 to 500 μg/ml based on the total weight of the composition, providing 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 provides a health functional food composition, 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 consumed for the purpose of improving vision through inhibition of glaucoma and macular degeneration, which are retinal diseases. The active ingredient may be ingested as a food prepared in tablets, capsules, powders, granules, pills, liquids, suspensions, or the like, or added to general foods. 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 drugs for a long time. The content of the active ingredient may be appropriately determined depending on the purpose of use (for prevention or improvement). In general, the active ingredient may be contained in 0.1 to 90% by weight in the health functional food composition. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the 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 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, gum, dairy products including ice cream, and various soups. , Beverages, alcoholic beverages and vitamin complexes, dairy and dairy products, etc., and health functional foods in the usual sense are included. The properties of the food are also not particularly limited, and may be solid, semi-solid, gel, liquid, powder, or the like.

It can be prepared as a beverage using the health functional food composition of the present invention. As an ingredient included in the beverage, there is no particular limitation on the selection of other ingredients other than the active ingredient, and various flavoring agents or natural carbohydrates, etc. may be included as an additional ingredient, as in ordinary beverages. Examples of the natural carbohydrates described above include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, and the like; And polysaccharides, for example, common 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 ratio 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, in addition to the active ingredient, the health functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavors and natural flavoring agents, coloring agents and thickeners (cheese, chocolate, etc.), pectic acid and Salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, etc. may be contained as additives. In addition, it may contain natural fruit juice and flesh 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 survival rate 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 diseases caused by oxidative stress.

Hereinafter, the present invention will be described in more detail through specific examples. The following examples are only examples to aid 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 farm in Hapcheon, Gyeongsangnam-do, and after crushing the outpost in the shade, 1 kg of the crushed centella asiatica was extracted with 50% alcohol for 5 hours at 80°C, concentrated and freeze-dried to 135 g of Gujeolcho extract (yield 13.5%) ).

[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).

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

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

As shown in FIG. 1, asiaticoside and asiatic acid compounds were detected in 50% centella asiatica extract, and were found to be 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 a DMEM/F12 medium containing 10% serum and antibiotics, and cultured in an incubator maintained at 37°C with a supply of 5 vol% of carbon dioxide. ARPE-19 cells were dispensed into a 96 well plate at a concentration of 5 × 10 ³ cells/well, and cultured for 24 hours. In order to measure the retinal pigment epithelial cell proliferation effect of asiaticoside and asiatic acid, after exchange with DMEM/F12 medium containing 10% charcoaled FBS, each compound was treated with 5, 10, 20 μg/mL, and positive As a control, 20 μM of lutein was additionally treated. After 48 hours, each cell growth rate was measured at an absorbance of 550 nm using an MTT reagent, and the absorbance of each test solution was measured based on the control group to indicate the cell proliferation rate (%).

The proliferation rates of retinal pigment epithelial cells of asiaticoside and asiatic acid are shown in FIG. 2.

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

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

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

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

The protective effect of asiaticoside and asiatic acid on oxidative damage to retinal cells is shown in FIG. 3.

As shown in FIG. 3, asiaticoside and asiatic acid exhibited superior protective effects against oxidative damage compared to the control group, and at 10 μg/mL concentration, asiaticoside was 21.5% compared to the control group treated with cobalt chloride, Asia Tic acid increased cell viability by 27.6%.

Experimental Example 4: Measurement of glucose metabolism efficacy in retinal epithelial cells

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

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

ARPE-19 cells were dispensed into 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 with 5, 10, 20 μg/mL, cultured for 24 hours, and then treated with serum-free DMEM/F12 medium and 100 μM of 2-NBDG for 30 minutes. I 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 efficacy of asiaticoside and asiatic acid is shown in FIG. 4.

As shown in FIG. 4, both asiaticoside and asiatic acid exhibited excellent glucose metabolism efficacy in retinal cells, and in particular, showed higher glucose metabolism than insulin, which is a positive control, so this result is asiaticoside and asiatic acid. It can be said that it is based on the retinal cell proliferation efficacy of

Accordingly, the present invention can provide a composition for preventing and 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.

[Preparation Example]

On the other hand, the pharmaceutical composition containing the active ingredient according to the present invention can be prepared in various forms depending on the purpose. Formulations 1 to 4 below are illustrative of a method for preparing a drug containing an active ingredient according to the present invention, and the present invention is not limited thereto.

Formulation 1: Tablet (direct pressure)

After sieving 5.0 mg of the active ingredient, 14.1 mg of lactose, 0.8 mg of crospovidone USNF, and 0.1 mg of magnesium stearate were mixed and pressed into 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 fine particles were pressed into tablets.

Formulation 3: powder and capsule

After sieving 5.0 mg of the active ingredient, 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate were mixed together. Mixture using a suitable device to solid No. Filled in 5 gelatin capsules.

Formulation 4: injection

An injection was prepared by containing 100 mg as an active ingredient, in addition to 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 containing the active ingredient according to the present invention can be prepared in various forms depending on the purpose. Formulations 5 to 9 below illustrate a method of preparing a health food containing an active ingredient according to the present invention, and the present invention is not limited thereto.

Formulation 5. Granular health food

Active ingredient 1000 mg, vitamin A acetate 70 ㎍, 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 ㎍, vitamin C 10 mg, biotin 10 ㎍, nicotinic acid amide 1.7 Mg, folic acid 50 ㎍, calcium pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, dicalcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate 100 Mg and 24.8 mg of magnesium chloride were mixed, and then granular health food was prepared according to a conventional method.

Formulation 6. Health drink

1000 mg of active ingredient, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, and 1 g of taurine 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 2 liter container, sealed and sterilized to prepare a health drink.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

Formulation 7. Flour food

0.5 to 5 g of the active ingredient was added to 100 g of wheat flour, and bread, cakes, cookies, crackers, and noodles were prepared using this mixture to prepare food for health promotion.

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. Sunsik

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

In the above, embodiments of the present invention have been described with reference to the accompanying drawings, but those of ordinary skill in the art to which the present invention pertains can be implemented in other specific forms without changing the technical spirit or essential features. You can understand that there is. Therefore, it should be understood that the embodiments described above are illustrative and non-limiting in all respects.

Claims (12)

An asiaticoside compound represented by the following 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 the following Formula 2, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof or a solvate thereof;
A composition for improving, preventing or treating diseases of retinal cell damage caused by oxidative stress, comprising any one or more of them as an active ingredient.
[Formula 1]

[Formula 2]

The method of claim 1,
The active ingredient is an asiaticoside compound represented by Formula 1, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate or a solvate thereof, to improve or prevent retinal cell damage diseases caused by oxidative stress. Or a therapeutic composition.
The method of claim 1,
The active ingredient may include an asiaticoside compound represented by Chemical 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 the following formula (2), an optical isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate or a solvate thereof; all are included, a disease of retinal cell damage caused by oxidative stress Composition for improvement, prevention or treatment of.
The method of claim 1,
The compound represented by Formula 1 and the compound represented by Formula 2 are each independently isolated or synthesized from Centella asiatica extract, a composition for improving, preventing or treating retinal cell damage diseases caused by oxidative stress.
The method of claim 1,
The compound represented by Formula 1 and the compound represented by Formula 2 are each independently isolated from centella asiatica extract, a composition for improving, preventing or treating retinal cell damage diseases caused by oxidative stress.
The method of claim 1,
The composition, characterized in that to improve the survival rate of retinal cells by increasing the sugar availability of the retinal cells, composition.
The method of claim 1,
The disease is a retinal disease macular degeneration or glaucoma, the composition.
The method according to any one of claims 1 to 7,
The composition for preventing or treating is a pharmaceutical composition, composition
The method according to any one of claims 1 to 7,
The composition for preventing or improving is a health functional food composition, composition
The method of claim 8,
The composition is administered in a dosage of 0.01mg / kg / day to 10g / kg / day, composition.
The method of claim 8,
The composition is a formulation for topical administration.
The method of claim 8,
The composition is an ophthalmic formulation.

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