KR102268002B1 - Sustained Release Eye-drop Composition - Google Patents

Abstract

The present invention relates to a long-acting eye drop composition comprising a DNA fraction and a biocompatible polymer. The ophthalmic composition of the present invention has low viscosity and excellent ophthalmic ophthalmic effect and prolonged drug release despite low viscosity, and exhibits both corneal and retina protective effects.

Description

Sustained Release Eye-drop Composition

The present invention relates to a long-acting eye drop composition. More specifically, the present invention is an eye drop composition comprising a DNA fraction and a biocompatible polymer, which has a low viscosity and a long duration of action of the drug, protects the cornea and retina, and has excellent pharmaceutical stability. is about

Tears consist of an oily layer, an aqueous layer and a mucous layer, and the tear film exhibits a three-layer structure. Since the concept of dry eye or dry eye (hereinafter referred to as 'dry eye') is broad, and the cause thereof is unknown in many cases, dry eye is not a single disease, but rather a decrease or fluctuation in the quantity or quality of tears. It is defined as an abnormal eye condition in which the tear film is unstable because the tear film is formed and the tear film disappears faster than normal. According to the above definition, the dry eye category includes dry corneal conjunctivitis, keratoconjunctival epithelial disorder, hypotonia, Stevens-Johnson syndrome, dry eye syndrome, Sjogren's syndrome, lacrimation syndrome, ocular hyperemia, tear film instability, or ophthalmic edema, etc. the same disease. Also, the category of dry eye includes allergic conjunctivitis, viral conjunctivitis, or dry eye after cataract surgery. In addition, with the recent increase in contact lens users, the increase in time spent in artificial air-conditioned environments, and the wide use of TVs and computers, the opportunity to view VDTs (Visual Display Terminals) has increased, which promotes dry eye. The factor increased significantly, and as a result, the category of dry eye includes contact lens wear-related dry eye or VDT work-related dry eye. Also, dry eye causes, in many cases, a keratoconjunctivitis disorder. In particular, when there is a lack of tears in the mucous layer, corneal damage is serious, which causes keratoconjunctival epithelial disorders, and the categories of keratoconjunctival epithelial disorders include dry eyes, corneal epithelial defects, conjunctival epithelial defects, corneal epithelial erosion, corneal thickness reduction, and corneas. Includes infiltration, corneal perforation, or corneal epithelial exfoliation, from keratoconjunctival epithelial disorders to corneal ulcer, keratitis, conjunctivitis, punctate keratosis, keratoconjunctivitis sicca, limbal keratoconjunctivitis, filamentous keratitis, corneal ulceration and infectivity of the corneal epithelium. eye diseases, etc. Corneal epithelial disorders can also be caused by intraocular trauma, microsurgery, or wearing hard contact lenses. Dry eye can occur not only in the normal eye, but also in the prosthetic eye. When the eyeball is lost due to various eye diseases, industrial accidents, or trauma to the eye including a traffic accident, the eyeball is removed and a prosthesis is worn, and patients who wear prosthetic eyes also complain of dry eye symptoms. Because.

If you feel dry eyes and discomfort, you can use an eye drop composition. In addition, when symptoms such as itching of the eyes or eyelids, congestion of the conjunctiva, general pain accompanied by burning eyes, glare, and tearing occur, eye drop compositions containing active ingredients (eg, antihistamines, etc.) Can be used.

The eye drop composition also contains preservatives such as benzalkonium chloride, chlorobutanol, and parahydroxybenzoate to prevent bacterial contamination. In several studies, these preservatives are toxic to corneal epithelial cells and epithelial. It has been reported to delay cell regeneration.

Preservative-free eye drop compositions without preservatives ^{typically include carboxymethylcellulose (eg Refresh Plus TM} ), and may also include hyaluronic acid as a polymer.

Numerous references are referenced throughout this specification and citations thereof are indicated. The disclosures of the cited documents are incorporated herein by reference in their entirety to more clearly describe the content of the present invention and the level of the art to which the present invention pertains.

Republic of Korea Patent Publication No. 10-2014-0041586

An object of the present invention is to provide an eye drop composition comprising a DNA fraction and a biocompatible polymer.

Another object of the present invention is to provide an eye drop composition having a low viscosity, a drug release pattern, and a protective effect on the cornea as well as the retina by using a DNA fraction and a biocompatible polymer in combination.

Further objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

One aspect of the present invention provides a composition comprising: (i) a DNA fraction; And (ii) to provide an eye drop composition comprising a biocompatible polymer selected from the group consisting of hyaluronic acid and salts thereof.

The DNA fraction is generally composed of biopolymers such as phosphoric acid, four types of bases, and deoxyribose, and is selected from polynucleotides (PN), polydeoxyribonucleotides (PDRN), and salts thereof. may be more than

The salt of PN or PDRN can be used without limitation as long as it is a pharmaceutically acceptable salt, which is a concentration having a relatively non-toxic and harmless effective action to the patient, and side effects due to this salt reduce the beneficial efficacy of PN and PDRN. Any and all organic or inorganic addition salts, suitable acid addition salts such as hydrochloride, bromate, iodate, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate , tartrate, pentothenate, bitartrate, ascorbate, succinic acid, maleate, gentisate, fumarate, gluconate, glucaronate, saccharide, formate, benzoate, glutamate, methanesulfonate , ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamonate salts, suitable base salts include, but are not limited to, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc and diethanolamine salts. it is not The salt may preferably be sodium polynucleotide or sodium polydeoxyribonucleotide.

The DNA fraction is known to have skin regeneration and corneal damage improvement effects, and is used as an active ingredient in water-soluble eye drops. When water-soluble eye drops are instilled, the instilled liquid is quickly removed from the cornea or conjunctiva by tears or blinking of the eyes. The effects of eye drops do not last long. On the other hand, long-acting eye drops that extend drug release time and residence time on the ocular surface affect not only the cornea but also the retina of the latter part of the eyeball, but there is no report on the effect of the DNA fraction on the retina.

Accordingly, the present inventors added a biocompatible polymer to the DNA fraction to obtain the effect of extending the drug's action time and enhancing moisture retention in addition to the known corneal protective effect of the DNA fraction. The present invention has revealed the surprising fact that, despite the low viscosity, excellent eye drop and drug release, and even the retinal protective effect that the DNA fraction alone does not have occurs due to the combination with the biocompatible polymer. was completed.

The eye drop according to the present invention contains one or more biocompatible polymers selected from the group consisting of hyaluronic acid and salts thereof along with the DNA fraction.

In the present invention, the 　 hyaluronic acid is a glycosaminoglycan that is an essential element of the extracellular matrix (ECM), and is a kind of mucopolysaccharide composed of amino acids and uronic acid, and the monomer N-acetylglucosamine ( It is a high molecular compound of a linear polysaccharide in which N-acetylglucosamine) and D-glucuronic acid are continuously linked. In addition, 　hyaluronic acid is a basic component of living tissue, an essential material for cell morphogenesis, cell differentiation, and cell division, and is a biocompatible material that helps heal wounds. Hyaluronic acid is insoluble in aqueous solution through an ether bond, but exhibits excellent viscoelasticity and high water absorption ability, so it is decomposed and absorbed in the body after maintaining its shape in the body for a certain period of time. Since natural hyaluronic acid is rapidly decomposed by hyaluronic acid degrading enzymes in the body, hyaluronic acid derivatives that are crosslinked in various ways or modified in structure using chemicals such as benzyl alcohol to control the degradation rate can also be used to create

In the present invention, the term "hyaluronic acid" includes not only natural hyaluronic acid itself, but also a hyaluronic acid derivative crosslinked by a crosslinking agent or a hyaluronic acid derivative whose structure is modified using a chemical substance such as benzyl alcohol. Concept. Hyaluronic acid may exist in a linear or cross-linked form, and the degree of cross-linking may be appropriately controlled by those skilled in the art. For example, although not limited thereto, those having an average molecular weight of 10 to 1,200 kDa may be used.

The 　 salt of hyaluronic acid may be in any salt form suitable for application to the body, and may be sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronate, and tetrabutylammonium hyaluronate. It may be one or more selected from the group consisting of. Sodium hyaluronate among the salts of hyaluronic acid is a highly lubricating hydrophilic polymer, and is described in Merck Index, 11th edition (Merck Index No.: 12, 4793) and the European Pharmacopoeia, 3rd Edition, (Supplement 2000, pages). 1190-1193, Appendix 3)] and the like, and the CAS number is CAS 9067-32-7. Although not limited thereto, those having an average molecular weight of 200 to 4,000 kDa, 750 to 2,000 kDa, 800 to 1,750 kDa, 900 to 1,500 kDa, or 1,000 kDa may be commonly used.

In one embodiment, the eye drop composition of the present invention may include 0.05 to 5.0% (w/v) of the DNA fraction and 0.001 to 0.010% (w/v) of the biocompatible polymer.

The eye drop composition according to the present invention may have a low viscosity and excellent eye drop. More specifically, low viscosity means a viscosity of 1 to 10 cP at 20° C., more preferably 1 to 5 cP, and most preferably 1 to 1.5 cP. Viscosity may be measured according to various methods known in the art, for example, a viscometer Brookfield Digital Viscometer, Model DV-1), Spindle No. 18 may be used, but is not limited thereto.

In the composition according to the present invention, in addition to the above-described DNA fraction and biocompatible polymer component, various components may be additionally included as long as the object of the present invention is not impaired.

For example, an ophthalmic polymer may be further included. The ophthalmic polymer includes glycerin, hydroxylpropylmethylcellulose, hydroxylethylcellulose, chondroitin sulfate, glucosamine, dextran, polysorbate 80 Polysorbate 80), polyethylene glycol 400, and glucose, but are not limited thereto.

In addition, the eye drop composition of the present invention may include additives such as, for example, a pharmaceutically acceptable carrier, excipient or diluent, and characteristics to be considered in the excipient include compatibility with other components included in the composition, biocompatibility, and the like. , processing temperature, etc. are included, but are not limited thereto.

In one embodiment, the eye drop composition according to the present invention can be prepared as an ophthalmic solution, in which case any dosage form used as an ophthalmic solution, such as an aqueous ophthalmic solution, an aqueous emulsion ophthalmic solution, a viscous ophthalmic solution and aqueous ophthalmic solutions such as dissolved ophthalmic solutions; or non-aqueous ophthalmic solutions such as non-aqueous ophthalmic solutions and non-aqueous emulsion ophthalmic solutions.

In addition, as long as the purpose of the present invention is not impaired, various additives known in the art may be included, for example, isotonic agents, buffers, stabilizers, pH adjusters, thickeners, preservatives, chelating agents, solubilizers and solvents, etc. may be included. .

The buffer may be selected from, but is not limited to, the group consisting of a phosphate buffer, a borate buffer, a citrate buffer, a tartrate buffer, an acetate buffer (eg, sodium acetate), tromethamine, and amino acids. Preferably, a phosphate buffer may be used.

The isotonic agent may be selected from the group consisting of, but not limited to, sorbitol, sugars such as glucose erythritol and mannitol, polyhydric alcohols such as glycerin, polyethylene glycol and polypropylene glycol, and salts such as sodium chloride, potassium chloride and magnesium chloride .

Preservatives include benzalkonium chloride, benzethonium chloride, alkyl paraoxybenzoates such as methyl paraoxybenzoate and ethyl paraoxybenzoate, benzyl alcohol, phenethyl alcohol, sorbic acid and its salts, thimerosal, polyquaternium , may be selected from the group consisting of benzododecinium bromide, oxychlorocomplex and chlorobutanol, but is not limited thereto.

Stabilizers include sodium edetate hydrate, cyclodextrin and derivatives thereof, water-soluble polymers such as poly(vinylpyrrolidone), and surfactants such as polysorbate 80 (Tween 80®), polysorbate 20, tyloxapol, troph It may be selected from the group consisting of methamine, but is not limited thereto, and sodium edetate hydrate may be preferably used.

The pH adjusting agent may be selected from the group consisting of hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, sodium hydroxide, potassium hydroxide, monoethanolamine, aqueous ammonia and ammonium hydroxide, but is not limited thereto.

The thickener may be selected from the group consisting of hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, hydroxypropylmethylcellulose and carboxymethylcellulose, carbomer, povidone, poloxamer, polycarbophil and salts thereof, but is limited thereto it doesn't happen

The chelating agent may be selected from, but not limited to, the group consisting of sodium edetate, sodium citrate and condensed sodium phosphate.

The solubilizing agent or solvent may be selected from glycerin, DMSO, DMA, N-methylpyrrolidone, ethanol, benzyl alcohol, isopropyl alcohol, polyethylene glycol or propylene glycol of various molecular weights, but is not limited thereto. There may be some overlap between components that can be used as solvents or solubilizers, and any component may be used as either a solvent or a solubilizer. If any component acts as a solvent in the formulation, it is reported as a solvent and the role of the solvent If it does not, it can be regarded as a solubilizer. Alternatively, the solubilizer may be a surfactant in some variations. Combinations of surfactants including various types of surfactants may be used. For example, nonionic, anionic (ie soap, sulfonate), cationic (ie CTAB), zwitterionic, polymeric, amphoteric surfactants can be used. For example, surfactants that may be used may include, but are not limited to, those having an HLB of 10, 11, 12, 13, or 14 or higher. Examples of surfactants are polyoxyethylene products of hydrogenated vegetable oils, polyethoxylated castor oil or polyethoxylated hydrogenated castor oil, polyoxyl castor oil or derivatives thereof, polyoxyethylene-sorbitan-fatty acid esters , polyoxyethylene castor oil derivatives and the like. However, the present invention is not limited thereto.

According to a specific embodiment, the eye drop composition of the present invention may include components such that the content ratio of the stabilizer and the DNA fraction is 1:8 to 1:16 (stabilizer: DNA fraction).

When the composition of the present invention is prepared as an emulsion ophthalmic solution, it may be prepared in a nanoemulsion type, and in this case, various additives known in the art may be included as long as the purpose of the present invention is not impaired, for example, oils and surfactants. etc. may be included. The oil is propylene glycol monocaprylate, propylene glycol laurate, medium chain (C8∼C10) triglycerides having 8 to 10 carbon atoms, glyceryl-1, At least one selected from the group consisting of 3-dioleate (glyceryl-1,3-dioleate), glyceryl monooleate (glyceryl monooleate), and glyceryl linoleate (glyceryl linoleate).

The eye drop composition according to the present invention can be used for improving, preventing or treating ophthalmic diseases, and preferably can be used for improving, preventing or treating disorders caused by dry eye.

The "dry eye disorders" include dry corneal conjunctivitis, keratoconjunctival epithelial disorders, hypotonia, Stevens-Johnson syndrome, dry eye syndrome, Sjogren's syndrome, lacrimation syndrome, ocular hyperemia, tear film instability, or ocular edema; dry eye after allergic conjunctivitis, viral conjunctivitis, or cataract surgery; and contact lens wear-related dry eye or VDT operation-related dry eye, and the like.

In addition, the "keratoconjunctival epithelial disorder" includes dry eye, corneal epithelial defect, conjunctival epithelial defect, corneal epithelial erosion, corneal thickness reduction, corneal infiltration, corneal perforation or corneal epithelial exfoliation; corneal ulcer, keratitis, conjunctivitis, superficial punctate keratosis, keratoconjunctivitis sicca, limbal keratoconjunctivitis, filamentous keratitis, corneal ulcer and infectious eye disease of the corneal epithelium; These may include intraocular trauma, microsurgery, or corneal epithelial disorders related to wearing hard contact lenses.

For the treatment and/or prophylaxis of a mammalian patient, particularly a human, the dosage of the composition according to the present invention can be routinely determined by a medical practitioner or a person skilled in the art. For example, when the composition according to the present invention is used as an eye drop for an adult dry eye patient, the preferred dosage of the composition is, for example, 1 to 4 drops per day (about 0.025 to 0.1 mL) of the eye drop, 1 to 1 per day. It may be administered up to 10 times, but is not limited thereto, and a medical practitioner or a person skilled in the art may determine the most suitable actual dosage according to the age, weight, sex and response of the patient to be treated, as well as the circumstances to be treated.

The composition of the present invention may include additional drugs (also referred to as “therapeutic agents” or “agents”) to ameliorate or treat the aforementioned ophthalmic diseases, such drugs, for example, glaucoma drugs (prostaglandin, latano Frost, etc.), muscarinic agents (pilocarpine, etc.), beta blockers (betaxolol, etc.), alpha agonists (brimonidine, etc.), carbonic anhydrase inhibitors (dorzolamide or brinzolamide, etc.), anti-inflammatory drugs (steroids, etc.) , soft steroids, or non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen), analgesics (such as salicylic acid and acetaminophen), antibiotics (such as beta-lactam antibiotics, macrocyclic antibiotics such as erythromycin, fluoroquinolone, etc.); antiviral agents (such as reverse transcriptase inhibitors or viral protease inhibitors), dry eye medications (such as cyclosporine, olapatadine, pain relievers, or sodium hyaluronate), and the like.

In a preferred embodiment, the eye drop composition of the present invention is a long-acting eye drop composition.

The term "persistence" refers to a characteristic of slowing the release rate of a drug when bioavailability is low, or when a drug is absorbed too slowly or is lost out of the body too quickly. According to this property, it exhibits an extended duration of action (action). Specifically, the eye drop composition comprising the DNA fraction and the biocompatible polymer according to the present invention has a drug release rate of 1.2 times or more, 1.5 times or more, 1.7 times or more, compared to an eye drop composition containing only a DNA fraction without a biocompatible polymer, Preferably, it may be 2 to 3 times or more (provided that in this case, the drug release rate is based on the time it takes for the long-acting eye drop composition to be homogenized in a balanced salt solution at pH 7.4 and 37°C).

The composition according to the present invention may be provided by filling in a sterile container, and may be provided including instructions on its use, and the instructions may be physically placed in a container filled with the composition or a second container packaging the container. It can be attached as a hood or packaged together inside a second container.

The ophthalmic composition of the present invention has low viscosity and excellent ophthalmic ophthalmic effect and prolonged drug release despite low viscosity, and exhibits both corneal and retina protective effects.

1 shows the results of evaluating the cell viability of retinal progenitor cells in order to evaluate the effect on the retina when the effect lasting eye drop is applied. PDRN alone does not have a protective effect on the retina, but cell proliferation is possible when used with natural polymer, and as a result, it was confirmed that it is most suitable as a long-acting eye drop.

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

Example

Preparation of eye drop composition

Each component and content were configured as shown in Table 1 below to prepare an eye drop composition.

purpose of mixing Input raw material name Raw material input (mg/mL) comparative example composition 1 composition 2 composition 3 composition 4 composition 5 composition 6 composition 7 composition 8 composition 9 composition 10 chief ingredient polydeoxyribo
sodium nucleotide 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 polymer Sodium Hyaluronate - 0.01 - - - - One - - - - carboxymethyl
Sodium Cellulose - - 0.01 - - - - One - - - polyvinylpyrrolidone - - - 0.1 - - - - 10 - - Polyethylenimine - - - - 0.1 - - - - 10 - gelatin - - - - - 0.01 - - - - One isotonic agent sodium chloride 0.9 0.9 0.9 0.9 0.9 0.9 0.9 0.9 0.9 0.9 0.9 pH regulator Phosphate Buffer appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount stabilizer Sodium edetate hydrate appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount solvent water for injection appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount appropriate amount

As a comparative example, an eye drop (BMI Korea, Korea) containing 0.75 mg of sodium polydeoxyribonucleotide per mL was purchased. Composition 1 is sodium polydeoxyribonucleotide, sodium hyaluronate, sodium chloride in 80 mL of sterile purified water. , Sodium hydrogen phosphate hydrate, sodium dihydrogen phosphate dihydrate, and sodium edetate hydrate were added and dissolved, sterilized purified water was added to make the total volume 100 mL, and then filtered with a 0.2 μm membrane filter.

Compositions 2 to 10 consisted of each component and content as shown in Table 1, and an eye drop composition was prepared in the same manner as in Composition 1.

Experimental example 1: downplay Stability test

It was observed whether precipitation occurred while the eye drop compositions prepared in Compositions 1 to 8 were stored at 40° C. and RH 75% and 2-8° C. (refrigerated) for 4 weeks, and the results are shown in Table 2 below.

Yongsi
(initial) 40℃, 75%RH 2-8℃ (refrigerated) 2 weeks 4 weeks 2 weeks 4 weeks comparative example transparent transparent transparent transparent transparent composition 1 transparent transparent transparent transparent transparent composition 2 transparent transparent transparent transparent transparent composition 3 transparent transparent transparent transparent transparent composition 4 transparent transparent transparent transparent opaque precipitation composition 5 transparent transparent transparent transparent Generates a small amount of transparent slurry composition 6 transparent transparent transparent transparent transparent composition 7 transparent transparent transparent transparent transparent composition 8 transparent transparent transparent transparent transparent composition 9 transparent transparent transparent opaque precipitation opaque precipitation composition 10 transparent transparent transparent Generates a small amount of transparent slurry Generates a small amount of transparent slurry

As a result of the experiment, as shown in Table 2, it was confirmed that all eye drops except for compositions 4, 5, 14, and 15 had excellent stability over time.

Experimental example 2: Viscosity measurement test

The viscosity and viscoelastic modulus ratio (G''/G') of the eye drops prepared according to Compositions 1 to 4 and Compositions 9 to 12 were measured, and the results are shown in Table 3. It was tested according to the Korean Pharmacopoeia general test method, and a viscometer Brookfield Digital Viscometer, Model DV-1 was used.

When the coefficient ratio is less than 1, that is, when G'>G'' is high, it can be said that a gel phase, which is an intermediate phase between liquid and solid, is obtained. As a result of the experiment, compared to other thickeners, it was judged that the viscosity of the eye drops was improved and close retention was possible by the anionic thickener.

Experimental Example 3: Drug release test

In order to confirm drug release of the eye drop compositions prepared according to Compositions 1 to 4 and Compositions 9 to 12, the test was conducted in the following manner. The drug release test solution of eye drops was tested at 37°C with a balanced salt solution (pH 7.4). A sample of the eye drop composition dyed with red No. 3 dye at a concentration of 5 mg/mL was carefully placed in a round-bottomed beaker containing 250 mL of a balanced salt solution, one drop each, while maintaining the temperature of the container at 37°C. The measured time taken for homogenization in the solution was taken as the drug release time, and the time was measured. The results are shown in Table 4.

Release time (sec) comparative example 24 composition 1 44 composition 2 42 composition 3 32 composition 4 30 composition 5 34 composition 6 78 composition 7 72 composition 8 55 composition 9 55 composition 10 56

As shown in Table 4, it was confirmed that the release time of the eye drops containing the biopolymer prepared in the composition of the present invention was extended by about 2 times or more compared to the Comparative Example that did not contain the biopolymer.

Experimental example 4: Retinal protection experiment

When eye drops such as artificial tears are instilled into the eye, they immediately affect the corneal epithelial cells. However, in the case of eye drops with a short residence time on the surface of the eye, they are easily removed by tears and have little effect on the retina located in the latter part of the eyeball. On the other hand, in the case of eye drops that extend the viscosity, drug release time, and residence time on the ocular surface, the effects are even on the retina.

Therefore, in order to evaluate the effect of eye drops on the retina during eye instillation, cultured retinal progenitor cells (RPCs) ^{were attached to each plate by 10 × 10 4} cells/well, and the medium in the upper layer was removed. The eye drops of Comparative Examples and Compositions 1 to 8 were dispensed and the effect on cells was tested. Specifically, cell morphology was observed on days 0, 3, and 7 after contact with eye drops, and cell viability was analyzed using a CCK-8 assay kit, and the results are shown in FIG. 1 and Table 6.

evaluation time Day 0 Day 3 Day 7 Cell growth rate (%) comparative example 98.8 99.4 101.1 composition 1 99.1 107.7 120.1 composition 2 98.4 94.4 92.4 composition 3 99.1 99.2 99.0 composition 4 99.4 93.2 82.8 composition 5 99.1 106.2 118.8 composition 6 98.8 114.1 120.9 composition 7 99.0 95.1 92.0 composition 8 99.2 99.0 97.7 composition 9 99.7 87.7 70.8 composition 10 98.9 117.1 122.1

1 and Table 6, in the case of Comparative Example 1 in which PDRN was used alone, no toxicity was observed, but a significant increase in cell growth rate was also not observed from day 0 to day 7, so that PDRN alone was not used in the retina. It was found to have no protective effect. On the other hand, when PDRN and hyaluronic acid (compositions 1 and 6), or PDRN and gelatin (compositions 5 and 10) were combined, an increase in cell growth rate by about 120% compared to the initial was observed. As described above, the composition in combination with the natural polymer and PDRN has been confirmed to have not only corneal protection but also a remarkable retinal protection effect, and thus is evaluated as the most suitable as eye drops, especially eye drops with long-lasting effects.

Experimental Example 5: Surface charge evaluation

The drug absorption and efficacy of artificial tears and eye drops used for dry eye syndrome are closely related to the passage of the phospholipid layer of the cornea at the outermost part of the eyeball. At this time, the phospholipid layer of the cornea has an anionic charge property, and based on this, it has been found that the composition applied to the cornea is advantageous in passing through the barrier when it has a cationic surface charge (Malhotra et al., Permeation through cornea). , Indian J Exp Biol. 39:1 (2001): 11-24). Therefore, in order to confirm the surface charge of the composition of the present invention, each test example was prepared as shown in Table 7, its zeta-potential value was measured, and the results are shown in Table 8. A Zetasizer Nano ZS (Malvern) instrument was used for the measurement.

purpose of mixing Input raw material name Raw material input (mg/mL) Comparative Example 1 Comparative Example 2 composition 1 chief ingredient polydeoxyribo
sodium nucleotide 0.75 0.75 polymer Sodium Hyaluronate - 0.01 0.01 carboxymethyl
Sodium Cellulose - - - polyvinylpyrrolidone - - - Polyethylenimine - - - gelatin - - - isotonic agent sodium chloride 0.9 0.9 0.9 pH regulator Phosphate Buffer appropriate amount appropriate amount appropriate amount stabilizer Sodium edetate hydrate appropriate amount appropriate amount appropriate amount solvent water for injection appropriate amount appropriate amount appropriate amount

Surface charge (Zeta-potential) water for injection -0.1 ± 0.01 Comparative Example 1 -3.0 ± 0.24 Comparative Example 2 -12.5 ± 0.98 composition 1 -5.2 ± 0.54

As a result of the measurement, polydeoxyribonucleotides have a characteristic of having an anionic surface charge, which is thought to adversely affect the passage of anions into the corneal lipid layer. On the other hand, when hyaluronic acid was added to polydeoxyribonucleotides, it was confirmed that the absolute value of zeta-potential was reduced to 1/2 despite the presence of the same anionic raw material in the composition. This is contrary to the expectation that when hyaluronic acid is added rather than polydeoxynucleotide alone, each charge value is summed to represent a greater negative charge, rather the intensity of the negative charge is offset, and thus the passage of the lipid layer on the corneal surface is expected to increase.

Claims (11)

(i) a DNA fraction; and (ii) at least one biocompatible polymer selected from the group consisting of hyaluronic acid and salts thereof, as a sustained release low-viscosity ophthalmic eye drop comprising:
The viscosity of the eye drops is 1 to 10 cP at 20 ℃ ophthalmic eye drops. The ophthalmic eye drop according to claim 1, wherein the DNA fraction is at least one selected from the group consisting of polynucleotides (PN), polydeoxyribonucleotides (PDRN), and salts thereof. The method of claim 1, wherein the hyaluronic acid salt is selected from the group consisting of sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronate, and tetrabutylammonium selected from the group consisting of hyaluronic acid. ophthalmic eye drops. According to claim 1, wherein the DNA fraction 0.05 to 5.0% (w / v) and the biocompatible polymer 0.001 to 0.010% (w / v) ophthalmic eye drop characterized in that it contains. The ophthalmic eye drop according to claim 1, wherein the viscosity of the eye drop is 1 to 5 cP. The ophthalmic eye drop according to claim 1, wherein the eye drop further comprises at least one adjuvant selected from the group consisting of an isotonic agent, a pH adjusting agent, and a stabilizer. The ophthalmic eye drop according to claim 6, wherein the isotonic agent is at least one selected from the group consisting of sodium chloride, mannitol, sorbitol, potassium chloride and magnesium chloride. The ophthalmic eye drop according to claim 6, wherein the stabilizing agent is sodium edetate hydrate. The ophthalmic eye drop according to claim 6, wherein the content ratio of the stabilizer and the DNA fraction is 1:8 to 1:16 (stabilizer: DNA fraction). delete The ophthalmic eye drop according to claim 1, wherein the eye drop has a corneal and retina protective effect.

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