KR102159223B1 - Composition for improving, preventing or treating corneal damage containing myonectin - Google Patents

Abstract

The present invention relates to a composition for alleviating, preventing or treating corneal damage using myonectin as an active ingredient. When the myonectin expressed in muscle is instilled in an animal model of corneal damage, it is confirmed that the death and inflammation of corneal epithelial cells are inhibited and treated. Accordingly, the composition using myonectin as an active ingredient can be utilized as the composition for alleviating, preventing or treating corneal damage.

Description

Composition for improving, preventing, or treating corneal damage containing myonectin as an active ingredient {Composition for improving, preventing or treating corneal damage containing myonectin}

The present invention relates to a composition for improving, preventing or treating corneal damage using myonectin as an active ingredient.

On the other hand, since the advent of personal computers in the 1980s, the continued use of eyes in the information society in which portable video devices including computers and popularized smartphones have been popularized has been further forced regardless of gender and age. The need for is becoming more important. In particular, amid the increase in the number of imaging media, the number of cases of complaining of dry eye symptoms, which are known to be common in the elderly, in young adults has been gradually increasing. In particular, the number of patients suffering corneal damage as a side effect of the recent increase in the use of contact lenses and the prolonged exposure to fine dust is increasing.

Myokine is an activator expressed in skeletal muscle during muscle contraction. Proteins and hormones secreted from muscle fibers directly affect muscle cells, the main disease tissue of muscle cells, or other tissues through blood flow. Affect Among them, myonectin is secreted by skeletal muscle to regulate systemic fatty acid metabolism, and is a member of the C1q/TNF-related protein (CTRP) family, and is known to be abundantly expressed in skeletal muscle tissue, especially type I muscle fibers. have. In particular, myonectin is released into the bloodstream by muscle contraction, and is known to promote fatty acid absorption into cells by increasing the expression of fatty acid transport genes (CD36, FATP1, Fabp1, and Fabp4). There is currently no known effect on the treatment of the disease.

Accordingly, the present inventors tried to find a new substance for treating corneal damage, and found that myonectin inhibits the death and inflammatory response of corneal epithelial cells due to corneal damage, thereby completing the present invention. .

Patent Document 1. Korean Registered Patent Publication No. 10-1734896

The present invention was conceived in view of the above problems, and an object of the present invention is to improve, prevent or treat corneal damage, including myonectin, which has an excellent therapeutic effect against damage to the cornea or corneal epithelial cells. It is intended to provide a composition.

The present invention for achieving the object as described above is to provide a pharmaceutical composition for preventing or treating corneal damage using myonectin as an active ingredient.

The corneal damage is Stevenson-Johnson syndrome, Sjogren syndrome, dry eye syndrome, trauma, eye trauma caused by eye surgery, eye trauma caused by chemicals, heat or radiation, infectious or non-infectious uveitis, immunity after corneal transplantation. It may be any one or more selected from the group consisting of exogenous diseases and corneal epithelial disorders caused by rejection reactions and wearing lenses.

The corneal injury may be an ocular trauma caused by chemicals, heat or radiation.

The myonectin may inhibit the expression or activity of MMP-2, MMP-9, or annexin V in corneal epithelial cells.

The composition may be an ocular external formulation.

The composition may further include a pharmaceutically acceptable carrier.

In addition, in order to achieve another object as described above, the present invention provides a food composition for improving or preventing corneal damage using myonectin as an active ingredient.

The corneal damage is Stevenson-Johnson syndrome, Sjogren syndrome, dry eye syndrome, trauma, eye trauma caused by eye surgery, eye trauma caused by chemicals, heat or radiation, infectious or non-infectious uveitis, immunity after corneal transplantation. It may be any one or more selected from the group consisting of exogenous diseases and corneal epithelial disorders caused by rejection reactions and wearing lenses.

The corneal injury may be an ocular trauma caused by chemicals, heat or radiation.

The myonectin may inhibit the expression or activity of MMP-2, MMP-9, or annexin V in corneal epithelial cells.

In addition, in order to achieve another object as described above, the present invention provides a contact lens or a composition for preserving an artificial lens containing myonectin as an active ingredient.

According to the present invention, when myonectin expressed in muscle was instilled in an animal model of corneal injury, it was confirmed that the death and inflammation of corneal epithelial cells were inhibited and treated. Therefore, a composition using myoneectin as an active ingredient can be utilized as a composition for improving, preventing or treating corneal damage.

1 is a diagram showing an experimental design for confirming the therapeutic effect of myonectin on corneal damage.
2 is a photograph of the results of bio-optical imaging according to whether mynectin is administered to an animal model of corneal injury.
3 is a graph showing a quantitative region of interest (ROI) analysis result obtained through the bio-optical imaging result of FIG. 2.

Hereinafter, a composition for improving, preventing or treating corneal damage using myonectin according to the present invention as an active ingredient will be described in detail.

In the present invention, myonectin is one of the new myokine proteins, and was discovered in 2012 (Marcus M. Seldin, Jonathan M. Peterson, Mardi S. Byerly, Zhikui Wei, and G. William Wong "Myonectin (CTRP15), a Novel Myokine That Links Skeletal Muscle to Systemic Lipid Homeostasis." THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 287, NO. 15, pp. 11968-11980, April 6, 2012, doi: 10.1074/jbc. M111.336834 originally published online February 17, 2012). Myoneectin is secreted by skeletal muscle to regulate systemic fatty acid metabolism, and is a member of the C1q/TNF-related protein (CTRP) family, and is known to be abundantly expressed in skeletal muscle tissue, especially type I muscle fibers.

Myoneectin is mainly expressed and secreted by skeletal muscle, and regulates nutrient metabolism secreted by skeletal muscle in response to changes in cellular energy state due to glucose, but it is known that it becomes difficult to control in obese state.

In the present invention, it was confirmed that the expression of MMP-2 and MMP-9 was decreased in corneal or corneal epithelial cells by treatment with myonectin in an animal model of corneal injury, thereby improving, preventing or preventing corneal damage of myonectin. The therapeutic effect was identified.

One aspect of the present invention provides a pharmaceutical composition for preventing or treating corneal damage using myonectin as an active ingredient.

In the present invention, the cornea is a transparent, blood vesselless tissue occupying 1/6 of the front eyeball surface, and not only protects the eye from the outside, but also plays a major function in refraction and transmission of light, and nerves are developed. The cornea is composed of five layers: corneal epithelium, Bowman's membrane, corneal parenchyma, desme's membrane, and corneal endothelium, and corneal epithelial cells occupy about 10% of the thickness of the entire cornea and are connected to the epithelium of the conjunctiva. It consists of 7 layers of cells, and the bottom cells of the lowermost layer proliferate and push outward and drop out after 7-14 days. The cornea is a thin tissue with a central thickness of 0.5 mm and is easily ruptured by severe impact.

Therefore, the corneal injury of the present invention is not particularly limited as long as symptoms of damage to the corneal tissue of the eyeball are accompanied, but specifically, Stevenson-Johnson syndrome, Sjogren syndrome, dry eye syndrome, trauma, eye trauma due to eye surgery, chemical substances , Eye trauma caused by heat or radiation, infectious or non-infectious uveitis, immune rejection after corneal transplantation, exogenous diseases caused by wearing lenses, and corneal epithelial disorders. . The eye surgery refers to any surgery for incision of the eyeball, and may be typically any one or more selected from the group consisting of cataract surgery, glaucoma surgery, retinal surgery, LASIK surgery, and LASEK surgery.

Preferably, the corneal damage may be an ocular trauma caused by chemicals, heat or radiation, and the chemical may be ethanol.

In the present invention, it was confirmed that the expression of MMP-2 and MMP-9 in the cornea was increased in the right eye of the corneal injury animal model in which the corneal injury was induced by instillation of ethanol compared to the normal left eye. In addition, when a probe capable of detecting MMP-2 and MMP-9 was administered, the experimental group to which myonectin was administered continued to express MMP-2 and MM-9 compared to the control group to which myonectin was not administered. Since it was confirmed that the decrease was confirmed, it can be determined that the myonectin of the present invention has an improvement, prevention or treatment effect on corneal damage.

In the present invention, "containing as an active ingredient" or "including as an active ingredient" means including an amount sufficient to improve, prevent or treat corneal damage of the myonectin of the present invention. Since the myonectin is a stable biomolecule and has no side effects to the human body even when an excessive amount is administered, the quantitative upper and lower limits of the myonectin contained in the composition of the present invention can be selected and performed within an appropriate range by a person skilled in the art.

As described above, it may be used as a pharmaceutical composition for preventing or treating corneal damage using myonectin as an active ingredient. It can be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient myonectin, and the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, lubricants or flavoring agents. Can be used.

For administration, the pharmaceutical composition may include one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Furthermore, it can be preferably formulated according to each disease or ingredient using a method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration and eye drop administration, and preferably transdermal administration. It is administration or instillation.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, and is usually As such, a skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.0001-10 g/kg, and more preferably 0.01 g/kg to 10 g/kg may be administered.

The pharmaceutical composition of the present invention may be prepared in a unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient, or may be prepared by placing it in a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

The pharmaceutical composition for preventing or treating corneal damage using myonectin as an active ingredient of the present invention may be formulated into an ocular external formulation by a conventional method by combining it with a carrier commonly accepted in the pharmaceutical field. It is preferable that the ocular external formulation is an eye drop.

The ocular external formulation is preferably an isotonic aqueous solution or suspension, and the composition mentioned is sterilized and/or contains an adjuvant (a preservative, a stabilizer, a wetting agent or a salt or buffer for controlling the osmotic pressure). In addition, they may contain other therapeutically useful substances.

As a general ocular external formulation, anionic polymers such as hyaluronic acid and carboxymethylcellulose, or pharmaceutically acceptable salts thereof, are known to have moisturizing and lubricating effects in eye drops, and in addition to these components, pharmaceutically acceptable carriers are used. Can include. Examples of the carrier include isotonic agents, buffers, stabilizers, pH adjusters, and solvents. The isotonic agent plays a role of stabilizing the external formulation for the eye, and in general, sodium edetate and sodium perborate may be used. The pH adjuster adjusts the pH of the external preparation for the eye, and hydrochloric acid or sodium hydroxide may be used.

As a solvent, it is preferable to use sterile purified water or distilled water for injection, and the external formulation for the eye is preferably a liquid formulation. In addition, preservatives, preservatives, etc. may be added as needed.

In the present invention, the recommended amount and the number of times of use of the external ocular formulation may be instilled 1 to 3 drops once, 5 to 6 times a day, and may be appropriately adjusted according to symptoms. The dosage level for a specific patient may vary depending on the patient's weight, age, sex, health status, administration time, frequency of administration, and severity of disease.

The composition of the present invention may be variously used as a food composition for improving and preventing corneal damage using myonectin of the present invention as an active ingredient. The food composition containing myonectin of the present invention as an active ingredient may be formulated in the same manner as the pharmaceutical composition described below to be used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, health supplements. Etc.

The food composition of the present invention may include not only myonectin as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. . Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts, etc. may be further included in addition to myonectin of the present invention. .

The present invention provides a health functional food comprising a food composition for improving, inhibiting or preventing corneal damage comprising the myonectin as an active ingredient. Health functional foods are foods prepared by adding myonectin to food ingredients such as beverages, teas, spices, chewing gums, confectionery, or encapsulated, powdered, or suspension. However, unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of myonectin added in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but it can be added within the range that does not damage the original taste of the food. It is in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, another aspect of the present invention relates to a composition for preserving a contact lens or an artificial lens containing myonectin as an active ingredient.

In the case of a composition for preserving contact lenses, in addition to myonectin, an aqueous solution for storing contact lenses may generally contain saline, other buffer solutions, or deionized water, preferably boron-based buffers such as boric acid and borax, acetic acid , Acetic acid buffers such as sodium acetate and potassium acetate, phosphate buffers such as sodium hydrogen phosphate, sodium dihydrogen phosphate and potassium dihydrogen phosphate, carbonate buffers such as sodium carbonate and sodium hydrogen carbonate, citric acid, citric acid such as sodium citrate Based buffers or tromethamol buffers. More preferably, it may contain a salt-containing brine containing sodium chloride, sodium borate, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate or a corresponding potassium salt thereof. In addition, wetting agents, surfactants, stabilizers, viscosity modifiers, tonicity agents, solubility aids, antioxidants, preservatives, refreshing agents, chelating agents or emollients may further be included.

The artificial lens is used to replace the original lens when the patient's lens is diseased or damaged, and is mainly implanted in the eye to replace the original lens removed from the eye during cataract surgery. Since the artificial lens is used in the human body, it is important to keep it safe from infection or contamination until implantation. Since the composition of the present invention exhibits a curative effect related to corneal damage, it is included in the artificial lens preservation solution to protect the artificial lens from external infection or contamination, and may act so as not to cause corneal damage when transplanted into the human body. The composition for preserving the artificial lens of the present invention may further include a wetting agent, an antibacterial agent, a stabilizer, an isotonic agent, a dissolution aid, a viscosity modifier, an antioxidant, or a buffer.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example 1. Verification of the effect of myonectin on corneal injury treatment in an animal model of corneal injury

1) Corneal injury animal model

First, in the right eye of a 10-week-old female BALB/c mouse, a paper disc soaked with 20% ethanol was instilled 3 times for about 30 seconds, and then PBS (phosphate-buffered saline) was instilled again and washed for 1 minute (irrigation). Thus, an animal model of corneal injury was produced.

Next, 100 µl of the MMP-2,9 probe (NpFlamma® MMP-2, 9 675) was injected through the tail vein in order to check the degree of corneal damage in real time into the corneal injury animal model to prepare a corneal injury animal model.

Thereafter, 10 µl of myonectin was instilled every 3 hours for 2 days (experimental group; myonectin). For the control experiment, PBS (Phosphate buffered saline), which is not a candidate material, was instilled (n=5 for each group).

After that, using the IVIS Imaging System (Ami X imaging system Spectral Instruments Imaging, AZ, USA) installed with a CCD camera (Xenogen, CA, USA) to check the degree of corneal damage in each group, 3, 6, 24, Taken at intervals of 48 hours. The above experimental process is shown in FIG. 1.

2) optical microscopy

Using the corneal injury animal model established in Example 1-1, the MMP-2,9 probe (NpFlamma® MMP-2, 9 675) signal in the cornea was measured with an optical microscope. More specifically, after inducing corneal damage and immediately administering MMP-2,9 probe (NpFlamma® MMP-2, 9 675) and myonectin, IVIS Imaging System (Ami) with CCD camera (Xenogen, CA, USA) installed. X imaging system Spectral Instruments Imaging, AZ, USA) was used, and the degree of corneal damage due to ethanol was confirmed by ROI.

FIG. 2 is a photograph of a bio-optical imaging result according to whether myonectin is administered to an animal model of corneal injury, and FIG. 3 is a graph showing a result of a region of interest (ROI) analysis obtained through the bio-optical imaging result of FIG. .

As shown in Fig. 2, a strong signal was observed by the MMP-2,9 probe (NpFlamma® MMP-2, 9 675) in the right eye inducing corneal damage, compared to the normal left eye of the control group. This means that the expression or activity of MMP-2,9 was increased in the right eye compared to the normal left eye of the control group.

In addition, in the case of the experimental group (myonectin) to which myonectin was administered after corneal damage was induced, the signal from the MMP-2,9 probe (NpFlamma® MMP-2, 9 675) compared to the control group to which myonectin was not administered. It was confirmed that was significantly reduced.

In addition, there was no difference in the degree of corneal damage between the left eye of the control group and the left eye of the experimental group, but after corneal damage was performed with ethanol, the signal from the MMP-2,9 probe (NpFlamma® MMP-2, 9 675) was remarkable. Increased.

As shown in Figure 3, looking at the ROI analysis results for the right eye of the control group and the experimental group, the degree of corneal damage in the right eye of the control group was continuously increased over time compared to the normal left eye, and the experimental group to which myonectin was administered (myonectin). It was confirmed that corneal damage did not increase, but rather decreased, and it was confirmed that the degree of corneal damage was significantly improved, prevented, or treated by administration of myonectin of the present invention after induction of corneal damage.

In the experimental group (myonectin) to which myonectin of the present invention was administered, the signal from the MMP-2,9 probe (NpFlamma® MMP-2, 9 675) was significantly reduced compared to the control group, which was found in corneal epithelial cells. It means that the expression or activity of MMP-2,9 is remarkably decreased. Since MMP-2,9 is a marker of corneal epithelial damage, it can be seen that myonectin according to the present invention can effectively treat corneal damage through its reduction.

Claims (11)

A pharmaceutical composition for preventing or treating corneal damage, using myonectin as an active ingredient. The method of claim 1,
The corneal damage is Stevenson-Johnson syndrome, Sjogren syndrome, dry eye syndrome, trauma, eye trauma caused by eye surgery, eye trauma caused by chemicals, heat or radiation, infectious or non-infectious uveitis, immunity after corneal transplantation. A pharmaceutical composition for preventing or treating corneal damage, characterized in that at least one selected from the group consisting of exogenous diseases and corneal epithelial disorders caused by rejection reactions and wearing lenses. The method of claim 1,
The corneal damage is a pharmaceutical composition for preventing or treating corneal damage, characterized in that the eye trauma caused by chemicals, heat or radiation. The method of claim 1,
The myonectin is a pharmaceutical composition for preventing or treating corneal damage, characterized in that it inhibits the expression or activity of MMP-2, MMP-9 or annexin V in corneal epithelial cells. The method of claim 1,
The composition is a pharmaceutical composition for preventing or treating corneal damage, characterized in that the external formulation for the eye. The method of claim 1,
The composition is a pharmaceutical composition for preventing or treating corneal damage, characterized in that it further comprises a pharmaceutically acceptable carrier. A food composition for improving or preventing corneal damage using myonectin as an active ingredient. The method of claim 7,
The corneal damage is Stevenson-Johnson syndrome, Sjogren syndrome, dry eye syndrome, trauma, eye trauma caused by eye surgery, eye trauma caused by chemicals, heat or radiation, infectious or non-infectious uveitis, immunity after corneal transplantation. Corneal damage improvement or prevention food composition, characterized in that at least one selected from the group consisting of exogenous diseases and corneal epithelial disorders caused by rejection reactions and wearing lenses. The method of claim 7,
The corneal damage is a food composition for improving or preventing corneal damage, characterized in that the eye trauma caused by chemicals, heat or radiation. The method of claim 7,
The myonectin is a food composition for improving or preventing corneal damage, characterized in that it inhibits the expression or activity of MMP-2, MMP-9 or annexin V in corneal epithelial cells. A composition for preserving contact lenses or artificial lenses containing myonectin as an active ingredient.

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