WO2012169849A2

WO2012169849A2 - Eye-drop composition for preventing or treating ocular diseases

Info

Publication number: WO2012169849A2
Application number: PCT/KR2012/004578
Authority: WO
Inventors: Sung-Tae Yoon; Jae-Kap JUN; Hueon-Sook YOU; Gi Hwan Kim
Original assignee: Huons Co., Ltd.
Priority date: 2011-06-10
Filing date: 2012-06-09
Publication date: 2012-12-13
Also published as: WO2012169849A3; CN103608021A; KR101134340B1

Abstract

Disclosed herein is an eye-drop composition for preventing or treating ocular diseases, which includes hyaluronic acid or a salt thereof, a magnesium ion, a sodium ion, a potassium ion and a chloride ion. The eye-drop composition may further include a phosphate ion, an aminocaproate ion, an edetate ion and a tetraborate ion. The eye-drop composition can effectively prevent or treat xerophthalmia and corneal damage, and can effectively prevent the propagation of bacteria for a long period of time, and can thus be suitably used several times over a predetermined period of time as well as a single time.

Description

EYE-DROP COMPOSITION FOR PREVENTING OR TREATING OCULAR DISEASES

The present invention relates to an eye-drop composition for preventing or treating ocular diseases, which includes hyaluronic acid or a salt thereof and ions, and which can be used as muti dose formulation.

The cornea is a transparent thin tissue having a thickness of about 1 mm and having no blood vessels, and is very important to the eyesight. The cornea is located at the front of the eyeball, and functions to prevent chemical invasions resulting from chemicals or biological invasions caused by microbes. A tear layer is provided on the epithelial layer of the cornea. The tear layer keeps an eye wet to prevent the adhesion of an eyelid and the conjunctiva and maintains the physiological state of the cornea or the conjunctiva constant.

The cornea and the conjunctiva are easily damaged because they are exposed to the outside, and, as a result, corneal and conjunctival epithelial tissues are easily damaged by various external and endogenous factors to cause disorders. For example, the disorder of corneal and conjunctival epithelial tissues can be caused by external factors, such as operations, chemicals, traumas, contact lens wearing and the like, and can also be caused by endogenous diseases, such as Sjogren’s syndrome, Stevens Johnson syndrome, xerophthalmia and the like.

Meanwhile, among the endogenous diseases that can cause disorders of the corneal and conjunctival epithelial tissues, xerophthalmia is an ocular disease caused by insufficient lubrication and the chronic lack of wetness of the eye. That is, xerophthalmia is a complex of various symptoms caused by the abnormality of a tear layer attributable to a reduction or change in the amount and quality of tears wetting an eye, is an ocular disease that is caused by various factors, and is called “keratoconjunctivitis”. Generally, xerophthalmia temporarily appears and then disappears, but, according to circumstances, continuously causes pain and foreign-body sensation. Therefore, if xerophthalmia is not properly treated, corneal damages, such as corneal epithelium erosion, corneal epithelium disorder, corneal ulcer and the like, can be caused, or eyesight will deteriorate.

In order to treat such a corneal and conjunctival epithelium disorder or xerophthalmia, conventionally, emollients such as artificial tear dosage form and the like, and drugs such as topical steroid, topical retinoid (for example: vitamin A), oral pilocarpine, topical pilocarpine, topical cyclosporine and the like have been used. However, generally, an emollient treatment method can mitigate xerophthalmia for a short period of time, but, in order to maintain the state in which the xerophthalmia is mitigated, an emollient must be frequently administered because the emollient cannot completely remove the physiological cause of xerophthalmia. Further, in treatment methods using drugs, the drug cannot remove or reduce the underlying cause of xerophthalmia, can threaten the health of a patient’s eye or can reduce the adaptability of a patient, and a combination of these factors can cause harmful side effects.

Further, the conventional emollients and drugs can cause damage to the eyeball because they include an antiseptic or the like. When they do not include an antiseptic, they cannot effectively prevent the proliferation and growth of microbes or the like, so that they are problematic in being used for a long period of time.

Therefore, it is required to develop an eye-drop composition which can effectively prevent or treat an ocular disease such as a corneal and conjunctival epithelium disorder, xerophthalmia or the like and which can be used for a long period of time by effectively preventing the propagation of microbes.

[Cited Reference]

[Patent Document]

KR 2008-0031195

Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide an eye-drop composition for preventing or treating ocular diseases, which includes hyaluronic acid or a salt thereof and various kinds of ions.

One aspect of the present invention provides an eye-drop composition for preventing or treating ocular diseases, including hyaluronic acid or a salt thereof, a magnesium ion, a sodium ion, a potassium ion, a chloride ion and, an aminocaproate ion, an edetate ion and a tetraborate ion.

Another aspect of the present invention provides an eye-drop composition for preventing or treating ocular diseases, including about 0.15 ~ about 0.2 wt% of hyaluronic acid or a salt thereof, about 0.25 ~ about 0.55 mmol/L of a magnesium ion, about 50 ~ about 80 mmol/L of a sodium ion, about 13 ~ about 18 mmol/L of a potassium ion, about 60 ~ about 75 mmol/L of a chloride ion and at least one selected from an aminocaproate ion, an edetate ion, a tetraborate ion and a phosphate ion.

Still another aspect of the present invention provides an eye-drop composition for preventing or treating ocular diseases, including about 0.15 ~ about 0.2 wt% of hyaluronic acid or a salt thereof, about 0.25 ~ about 0.55 mmol/L of a magnesium ion, about 50 ~ about 80 mmol/L of a sodium ion, about 13 ~ about 18 mmol/L of a potassium ion, about 60 ~ about 75 mmol/L of a chloride ion, about 0.8 ~ about 8.5 mmol/L of an aminocaproate ion, about 0.5 ~ about 3.5 mmol/L of an edetate ion and about 0.15 ~ about 1.5 mmol/L of a tetraborate ion.

Further still another aspect of the present invention provides an eye-drop composition for preventing or treating ocular diseases, including about 0.1 ~ about 0.3 wt% of a sodium hyaluronate, about 0.1 ~ about 0.6 mmol/L of a magnesium ion, about 40 ~ about 90 mmol/L of a sodium ion, about 10 ~ about 20 mmol/L of a potassium ion, about 40 ~ about 90 mmol/L of a chloride ion, about 0.5 ~ about 10 mmol/L of an aminocaproate ion, about 0.3 ~ about 7 mmol/L of an edetate ion and about 0.1 ~ about 3 mmol/L of a tetraborate ion.

Further still another aspect of the present invention provides an eye-drop composition for preventing or treating ocular diseases, which is prepared by mixing sodium hyaluronate, magnesium chloride hydrate, sodium chloride, potassium chloride, borax, aminocaproic acid, sodium edetate hydrate and water.

Further still another aspect of the present invention provides a method of preventing or treating ocular diseases by administrating an eye-drop composition which includes a hyaluronic acid or a salt thereof, a magnesium ion, a sodium ion, a potassium ion, a chloride ion, an aminocaproate ion, an edetate ion and a tetraborate ion.

The eye-drop composition for preventing or treating ocular diseases according to the present invention can effectively prevent or treat xerophthalmia and corneal damages, and can effectively prevent the proliferation and growth of bacteria for a long period of time, and can thus be suitably used several times for a predetermined period of time as well as one time.

The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which:

FIG. 1 is a view showing regions of right and left eyes of an examinee to observe a staining intensity of a cornea of each of the right and left eyes; and

FIG. 2 is a graph showing degree of satisfaction of an examinee after the administration of an eye-drop composition of the present invention.

The present invention provides an eye-drop composition for preventing or treating ocular diseases, which includes hydrophilic hyaluronic acid or a salt thereof, a magnesium ion (Mg²⁺), a sodium ion (Na⁺), a potassium ion (K⁺) and a chloride ion (Cl^-).

The eye-drop composition may further include an aminocaproate ion (NH₂(CH₂)₅COO^-), an edetate ion (C₁₀H₁₄N₂O₈ ^2-), a tetraborate ion (B₄O₇ ^2-), a phosphate ion or a mixture thereof. The phosphate ion may be represented by PO₄ ^3-, HPO₄ ^2- or H₂PO_4-, and preferably HPO₄ ^2-.

Hyaluronic acid is glycosaminoglycogen, and is a biocompatible material which is bonded with collagen and elastin to form cartilage tissues and which is present in epithelial tissues or neural tissues. Further, hyaluronic acid is a material which strengthens and softens joints, and was tested to treat rheumatic arthritis and osteoarthritis to acquire good results with respect to treatment of rheumatic arthritis and osteoarthritis. Further, hyaluronic acid is biocompatible, and is a major component of an extracellular matrix, which is known to mechanically protect and buffer iris tissues, retinal tissues, corneal cells, endothelial cells and epithelial cells because hyaluronic acid is widely distributed in the intercellular cement in bonding tissues to form an adhesive and elastic solution under a physiological condition.

Since the eye-drop composition including such hyaluronic acid is biocompatible and has proper adhesiveness, the residence time of the composition including hyaluronic acid is long in the eyes, thus keeping the eyes wet for a long period of time.

The eye-drop composition of the present invention includes about 0.15 ~ about 0.2 wt% of hyaluronic acid or a salt thereof, about 0.25 ~ about 0.55 mmol/L of a magnesium ion (Mg²⁺), about 50 ~ about 80 mmol/L of a sodium ion (Na⁺), about 13 ~ about 18 mmol/L of a potassium ion (K⁺), about 60 ~ about 75 mmol/L of a chloride ion (Cl^-), and at least one of an aminocaproate ion (NH₂(CH₂)₅COO^-), an edetate ion (C₁₀H₁₄N₂O₈ ^2-), a tetraborate ion (B₄O₇ ^2-), and a phosphate ion (PO₄ ^3-, HPO₄ ^2- or H₂PO_4-).

The eye-drop composition for preventing or treating ocular diseases according to the present invention, which includes hyaluronic acid or a salt thereof, can effectively prevent or treat xerophthalmia and corneal damage and can effectively prevent proliferation and growth of microbes for a long period of time by suitably adjusting the amounts of hyaluronic acid or a salt thereof and various kinds of ions, and thus can be suitably used as a multi dose formulation for a predetermined period of time as well as a single dose formulation. Particularly, the eye-drop composition of the present invention can effectively prevent or treat xerophthalmia and corneal damage and can effectively inhibit proliferation and growth of bacteria for a long period of time without using a calcium ion.

The eye-drop composition of the present invention can effectively prevent the propagation of Aspergillus niger, Candida albican, Staphylococcos aureus, Pseudomonas aeruginosa or Escherichia coli, and particularly Staphylococcos aureus, Pseudomonas aeruginosa or Escherichia coli.

The eye-drop composition of the present invention does not form a precipitate for a long period of time, and is not easily volatilized and thus remains in an eye for a long time because it has a proper viscosity of about 15 ~ about 24 dl/g, thereby effectively controlling the supply of moisture and the ion concentration of the eye.

Further, the eye-drop composition of the present invention satisfying all the required physical properties of pH 7.2 ~ 7.4, an osmotic pressure of about 140~about 160 mOsm/L and a limiting viscosity of about 15 ~ about 24 dL/g, does not cause a stinging pain nor a foreign-body sensation in the eyes.

The eye-drop composition of the present invention can exhibit the required physical property of a pH 7.2 ~ 7.4 without using a strong acid (hydrochloric acid) or a strong base (sodium hydroxide), thus effectively preventing a stinging pain when the eye-drop composition is applied to the eyes.

The eye-drop composition of the present invention may include about 0.15 ~ about 0.2 wt% of hyaluronic acid or the salt thereof, about 0.25 ~ about 0.55 mmol/L of the magnesium ion, about 50 ~ about 80 mmol/L of the sodium ion, about 13 ~ about 18 mmol/L of the potassium ion, about 60 ~ about 75 mmol/L of the chloride ion, about 8.5 mmol/L or less of the aminocaproate ion (NH₂(CH₂)₅COO^-), about 3.5 mmol/L or less of the edetate ion and about 1.5 mmol/L or less of the tetraborate ion (B₄O₇ ^2-).

Preferably, the eye-drop composition of the present invention may include about 0.15 ~ about 0.2 wt% of hyaluronic acid or the salt thereof, about 0.25 ~ about 0.55 mmol/L of the magnesium ion, about 50 ~ about 80 mmol/L of the sodium ion, about 13 ~ about 18 mmol/L of the potassium ion, about 60 ~ about 75 mmol/L of the chloride ion, about 0.8 ~ about 8.5 mmol/L of the aminocaproate ion (NH₂(CH₂)₅COO^-), about 0.5 ~ about 3.5 mmol/L of the edetate ion, and about 0.15 ~ about 1.5 mmol/L of the tetraborate ion (B₄O₇ ^2-).

More preferably, the eye-drop composition of the present invention may include about 0.15 ~ about 0.2 wt% of hyaluronic acid or the salt thereof, about 0.25 ~ about 0.55 mmol/L of the magnesium ion, about 50 ~ about 80 mmol/L of the sodium ion, about 13 ~ about 18 mmol/L of the potassium ion, about 60 ~ about 75 mmol/L of the chloride ion, about 0.8 ~ about 8.5 mmol/L of the aminocaproate ion (NH₂(CH₂)₅COO^-), about 0.5 ~ about 3.5 mmol/L of the edetate ion, about 0.15 ~ about 1.5 mmol/L of the tetraborate ion (B₄O₇ ^2-), and about 0.5 ~ about 5 mmol/L of the phosphate ion.

The composition including the hyaluronic acid or the salt thereof and the specific kinds of ions in the above amounts can effectively prevent or treat xerophthalmia and corneal damage, and can effectively prevent proliferation and growth of microbes, and thus can be suitably used several times over a long period of time.

In accordance with the eye-drop composition of the present invention, the salt of hyaluronic acid is not particularly limited, but preferably may be sodium hyaluronate.

The weight-average molecular weight of hyaluronic acid or the salt thereof may be suitably adjusted to such a degree that the eye-drop composition can be imparted with proper viscosity, but preferably may be about 800,000 ~ about 1,600,000.

The eye-drop composition can effectively prevent or treat a corneal and conjunctival epithelium disorder, xerophthalmia or corneal damages, which are caused by various factors.

For example, the eye-drop composition of the present invention may be used to prevent or treat disorders of corneal and conjunctival epithelial tissues, which are caused by external factors, such as operations, chemicals, traumas, wearing contact lenses and the like, and which is caused by endogenous diseases, such as Sjogren’s syndrome, Stevens Johnson syndrome, xerophthalmia and the like.

The amount of the eye-drop composition in the eyes may be suitably adjusted depending on the degree of symptoms. The eye-drop composition may be administered into the eyes twice to seven times a day by one drop to three drops once, and preferably three times to six times a day by one drop to three drops once.

The present invention provides an eye-drop composition including about 0.1 ~ about 0.3 wt% of sodium hyaluronate, about 0.1 ~ about 0.6 mmol/L of a magnesium ion (Mg²⁺), about 40 ~ about 90 mmol/L of a sodium ion (Na⁺), about 10 ~ about 20 mmol/L of a potassium ion (K⁺), about 40 ~ about 90 mmol/L of a chloride ion (Cl^-), about 0.5 ~ about 10 mmol/L of an aminocaproate ion (NH₂(CH₂)₅COO^-), about 0.3 ~ about 7 mmol/L of an edetate ion (C₁₀H₁₄N₂O₈ ^2-), and about 0.1 ~ about 3 mmol/L of a tetraborate ion (B₄O₇ ^2-).

In accordance with the present invention, the eye drop composition may include about 0.3 ~ about 7 mmol/L of a phosphate ion (PO₄ ^3-, HPO₄ ^2- or H₂PO_4-).

The present invention provides an eye-drop composition prepared by mixing sodium hyaluronate, magnesium chloride hydrate (MgCl₂·6H₂O), sodium chloride (NaCl), potassium chloride (KCl), borax (Na₂B₄O₇·10H₂O), aminocaproic acid (C₆H₁₃NO₂), sodium edetate hydrate (C₁₀H₁₄N₂Na₂O_8·2H₂O) and water.

In accordance with the present invention, the eye drop composition may be prepared by further adding and mixing sodium hydrogenphosphate hydrate (Na₂HPO₄·12H₂O).

In accordance with the present invention, the eye drop composition may be prepared by mixing about 0.1 ~ about 0.3 wt% sodium hyaluronate, about 0.08~about 0.2mg/mL of magnesium chloride hydrate, about 2.5~about 3.5mg/mL of sodium chloride, about 0.5~about 1.5mg/mL of potassium chloride, about 0.05~about 1mg/mL of borax, about 0.1~about 1.5mg/mL of aminocaproic acid, about 0.1~about 0.5mg/mL of sodium edetate hydrate and water.

In accordance with the present invention, the eye drop composition may be prepared by further adding and mixing about 0.8~about 1.5mg/mL of sodium hydrogenphosphate hydrate.

The present invention provides a method of preventing or treating ocular diseases by administrating an eye-drop composition which includes hyaluronic acid or a salt thereof, a magnesium ion, a sodium ion, a potassium ion, a chloride ion and at least one of an aminocaproate ion (NH₂(CH₂)₅COO^-), an edetate ion (C₁₀H₁₄N₂O₈ ^2-), a tetraborate ion (B₄O₇ ^2-) and a phosphate ion (PO₄ ^3-, HPO₄ ^2- or H₂PO_4-).

In accordance with the present invention, the method of preventing or treating ocular disease may include administrating the eye drop composition which contains about 0.1 ~ about 0.3 wt% of hyaluronic acid or the salt thereof, about 0.1 ~ about 0.6 mmol/L of the magnesium ion, about 40 ~ about 90 mmol/L of the sodium ion, about 10 ~ about 20 mmol/L of the potassium ion, about 40 ~ about 90 mmol/L of the chloride ion, about 0.5 ~ about 10 mmol/L of the aminocaproate ion, about 0.3 ~ about 7 mmol/L of the edetate ion and about 0.1 ~ about 3 mmol/L of the tetraborate ion.

In accordance with the present invention, the method of preventing or treating ocular disease may include administrating the eye drop composition which contains about 0.1 ~ about 0.3 wt% of hyaluronic acid or the salt thereof, about 0.1 ~ about 0.6 mmol/L of the magnesium ion, about 40 ~ about 90 mmol/L of the sodium ion, about 10 ~ about 20 mmol/L of the potassium ion, about 40 ~ about 90 mmol/L of the chloride ion, about 0.5 ~ about 10 mmol/L of the aminocaproate ion, about 0.3 ~ about 7 mmol/L of the edetate ion, about 0.1 ~ about 3 mmol/L of the tetraborate ion and about 0.3 ~ about 7 mmol/L of the phosphate ion.

In accordance with the present invention, the method of preventing or treating ocular disease may include administrating the eye drop composition which contains about 0.15 ~ about 0.2 wt% of hyaluronic acid or the salt thereof, about 0.25 ~ about 0.55 mmol/L of the magnesium ion, about 50 ~ about 80 mmol/L of the sodium ion, about 13 ~ about 18 mmol/L of the potassium ion, about 60 ~ about 75 mmol/L of the chloride ion, about 0.8 ~ about 8.5 mmol/L of the aminocaproate ion, about 0.5 ~ about 3.5 mmol/L of the edetate ion and about 0.15 ~ about 1.5 mmol/L of the tetraborate ion.

In accordance with the present invention, the method of preventing or treating ocular disease may include administrating the eye drop composition which contains about 0.15 ~ about 0.2 wt% of hyaluronic acid or the salt thereof, about 0.25 ~ about 0.55 mmol/L of the magnesium ion, about 50 ~ about 80 mmol/L of the sodium ion, about 13 ~ about 18 mmol/L of the potassium ion, about 60 ~ about 75 mmol/L of the chloride ion, about 0.8 ~ about 8.5 mmol/L of the aminocaproate ion, about 0.5 ~ about 3.5 mmol/L of the edetate ion, about 0.15 ~ about 1.5 mmol/L of the tetraborate ion and about 0.5 ~ about 5 mmol/L of the phosphate ion.

Hereinafter, the present invention will be described in more detail with reference to the following Examples. However, these Examples are set forth to illustrate the invention, and the scope of the present invention is not limited thereto.

In the following reagents and solvents, sodium hyaluronate was manufactured by LG Life Sciences Ltd., a magnesium chloride hydrate was manufactured by Daejung Chemicals & Metals Co., Ltd., potassium chloride was manufactured by BMP, BULK Medicines Pharmaceutics Ltd., a sodium hydrogenphosphate hydrate was manufactured by Daejung Chemicals & Metals Co., Ltd., sodium citrate hydrate was manufactured by Whawon Pharmaceutics Ltd., borax was manufactured by Merck & Co., Ltd., aminocaproic acid was manufactured by Whawon Pharmaceutics Ltd., sodium edetate hydrate was manufactured by Samchun Pure Chemicals Co., Ltd., and sterilized purified water was manufactured by Huons Co., Ltd.

Sodium hyaluronate having a weight-average molecular weight of 1,000,000, magnesium chloride hydrate (MgCl₂·6H₂O), sodium chloride (NaCl), potassium chloride (KCl), sodium hydrogenphosphate hydrate (Na₂HPO₄·12H₂O), borax (Na₂B₄O₇·10H2O), aminocaproic acid (C₆H₁₃NO₂), and sodium edetate hydrate (C₁₀H₁₄N₂Na₂O₈·2H₂O) were completely stirred and dissolved in 80 mL of sterilized purified water in the amounts given Example 1 of Table 1 below to prepare a mixed solution. Subsequently, sterilized purified water was added to the mixed solution such that the total volume of the mixed solution is 100 mL, and then the mixed solution was filtered by a membrane filter having a thickness of 0.2 μm to obtain an eye-drop composition 1.

An eye-drop composition 2 was obtained in the same manner as in Example 1, except that the above components were added in the amounts given in Example 2 of Table 1 below.

An eye-drop composition 3 was obtained in the same manner as in Example 1, except that the above components were added in the amounts given in Example 3 of Table 1 below.

An eye-drop composition 4 was obtained in the same manner as in Example 1, except that a sodium hydrogenphosphate hydrate (Na₂HPO₄·12H₂O) was not used, and that the above components were added in the amounts given in Example 4 of Table 1 below.

An eye-drop composition 5 was obtained in the same manner as in Example 1, except that the above components were added in the amounts given in Example 5 of Table 1 below.

A comparative eye-drop composition was prepared as follows such that it has the same components and concentration as Vismed eye drops (manufactured by Dupama Consulting Co., Ltd.)

Sodium hyaluronate having a weight-average molecular weight of 1,000,000, magnesium chloride hydrate (MgCl₂·6H₂O), calcium chloride hydrate (CaCl₂·2H₂0), sodium chloride (NaCl), potassium chloride (KCl), sodium hydrogenphosphate hydrate (Na₂HPO₄·12H₂O), and sodium citrate hydrate (C₆H₅Na₃O₇·2H₂O) were completely stirred and dissolved in 80 mL of sterilized purified water in the amounts given Comparative Example of Table 1 below to prepare a mixed solution. Subsequently, sterilized purified water was added to the mixed solution such that the total volume of the mixed solution is 100 mL, and then the mixed solution was filtered by a membrane filter having a thickness of 0.2 μm to obtain a comparative eye-drop composition.

[Table 1]

The eye-drop compositions 1 to 5 prepared in Examples 1 to 5 were stored for 6 months under room temperature conditions of 25±2°C and a relative humidity of 60±5% and under harsh conditions of 40±2°C and a relative humidity of 75±5%, and simultaneously whether or not the eye-drop compositions 1 to 5 had formed a precipitate under the same conditions was observed. The results thereof are given Table 2 below.

[Table 2]

As shown in Table 2 above, the eye-drop compositions 1 to 5 prepared in Examples 1 to 5 did not form a precipitate under harsh conditions of high temperature and high humidity nor under room temperature conditions. From the results, it is confirmed that the eye-drop compositions of the present invention may have high stability for a long period of time.

The pH, osmotic pressure and viscosity of each of the eye-drop compositions 1 to 5 prepared in Examples 1 to 5 were measured, and the results thereof are given Table 3 below. Here, Sevenmulti (Mettle Toledo) was used as a pH-meter, Osmomat 030-D (Gonotec) was used as an osmotic pressure meter, and the limiting viscosity thereof was measured by a capillary viscometer that was used in the first viscosity measuring method of the general test method of the Korean Pharmacopoeia.

[Table 3]

As shown in Table 3, all the pH, osmotic pressure and viscosity of each of the eye-drop compositions 1 to 5 satisfied the reference conditions of eye-drop compositions.

From the results, it is confirmed that the eye-drop compositions of the present invention have physical properties suitable for eye-drops.

Meanwhile, in the case of the comparative eye-drop composition, it is confirmed that the comparative eye-drop composition has a pH of 8.6 which is basic, so that it is required to properly adjust the pH thereof by adding an acidic material.

The growth or proliferation of microbes was observed by comparing the microbiological characteristics of the eye-drop compositions 1 to 5 prepared in Examples 1 to 5 with those of the comparative eye-drop composition for a predetermined amount of time.

Experiments were carried out using five kinds of experimental microorganisms. As the five kinds of experimental microorganisms, two kinds of fungi of Aspergillus niger (ATCC 16404) and Candida albicans (ATCC 10231) and three kinds of bacteria of Staphylococcos aureus (ATCC 6538), Pseudomonas aeruginosa (ATCC 9027) and Escherichia coli (ATCC 8739) were used. The eye-drop compositions 1 to 5 were inoculated with the five kinds of test microorganisms such that a high-concentration experimental group includes 10⁵ ~ 10⁶ experimental microorganisms per mL and a low-concentration experimental group includes 10³ experimental microorganisms per mL, and were then stored at a temperature of 20 ~ 25°C. After the inoculation, samples were extracted at each of 1, 2, 3, 6 and 7 days, respectively, and diluted to 1 mL to obtain diluted samples of 100 cfu/mL. Subsequently, 1mL of each of the diluted samples was inoculated on a plate, and was then placed in a culture medium of 20-25mL (a fungus was placed in a TSA (Tryptic soybean agar) culture medium, and a bacterium was placed in a SDA (Sabour dextrose agar) culture medium) and then cultured. Each of the plates inoculated with fungi was cultured at 20 ~ 25°C for five days, and each of the plates inoculated with bacteria was cultured at 30 ~ 35°C for three days.

The number (cfu (colony forming unit)) of microorganisms after culturing was examined with the naked eye, and the results thereof are given in Tables 4 to 8 below.

[Table 4]

[Table 5]

[Table 6]

[Table 7]

[Table 8]

[Table 9]

As shown in Tables 4 to 9, the number of microorganisms in the culture medium which each of the eye-drop compositions 1 to 5 was inoculated was decreased or maintained constant for 7 days, whereas the number of microorganisms in the culture medium which the comparative eye-drop composition was inoculated was greatly increased. Particularly, when the microorganism was Pseudomonas aeruginosa, the number thereof increased from 9x10⁶ to 6.8x10⁷ which is an increase of 10 times or more in the high-concentration experimental group, and the number thereof increased from 9.0 x10² to 8.4x10⁶ which is an increase of 10000 times or more, in the low-concentration experimental group.

From the results, it is confirmed that the eye-drop composition of the present invention may effectively control the proliferation and/or growth of microorganisms for a predetermined amount of time, and thus prevent eyes from being contaminated by bacteria for a long period of time, so that it can be used multi dose as well as a single dose.

The eye-drop tests (easiness in administration, stinging pain , foreign-body sensation) of 20 normal persons (10 men and 10 women) were carried out using the eye-drop compositions 1 and 2 prepared in Examples 1 and 2 and the comparative eye-drop composition having a pH of 7.3 which was obtained by the addition of hydrochloric acid and satisfied the reference condition of eye-drops. The results of the eye drop tests were scored according to score criteria, and are given in Tables 10 to 12 below.

[Table 10]

As shown in Table 10 above, the eye-drop compositions 1 and 2 of Examples 1 and 2 were comfortably and easily administered into the eyes because 90% or more of men and women assigned scores of 3 points or more, whereas the comparative eye-drop composition was not easily administered into eyes because 30% or more of men and women assigned scores of 2 points or less.

From the results, it is confirmed that the drop-eye composition of the present invention can be comfortably and easily administered into the eyes.

[Table 11]

As shown in Table 11 above, the eye-drop compositions 1 and 2 of Examples 1 and 2 barely stung when they were administered into eyes because 90% or more of men and women assigned scores of 3 points or more, whereas the comparative eye-drop composition stung when it was administered into eyes because 40% or more of men and women assigned scores of 2 points or less.

From the results, it is confirmed that the drop-eye composition of the present invention does not sting when administered into the eyes.

[Table 12]

As shown in Table 12 above, the eye-drop compositions 1 and 2 of Examples 1 and 2 did not create foreign-body sensation at all when they were administered into the eyes because 100% of men and women assigned scores of 3 points or more, whereas the comparative eye-drop composition created a foreign body sensation when it was administered into the eyes because about 10% of men and women assigned scores of 2 points or less.

From the results, it is confirmed that the drop-eye composition of the present invention does not create a foreign-body sensation at all when administered into the eyes.

Consequently, it is confirmed that the eye-drop composition of the present invention is easily administered into the eyes, does not sting and does not create a foreign-body sensation when it is administered into the eyes.

<Experimental Example 5> Evaluation of degree of alleviation of xerophthalmia

The eye-drop composition 1 prepared in Example 1 was administered into the eyes of examinees having mild or serious xerophthalmia (14 men and 39 women), and the treatment effect thereof was observed.

1. Evaluation of the state of eyes of examinees

Prior to administration of the eye-drop composition of the present invention, the state of eyes of examinees was evaluated as follows.

(1) Measurement of tear breakup time

A tear breakup time (TBUT) of examinees was measured by the following method.

An examinee sat with his head supported by a headrest. A slit lamp microscope was set at a magnifying power of 10, and was placed such that the angle of the microscope to illumination is 60˚. The slit lamp was adjusted such that the distance between slits was 3 mm, and the tear break up time (TBUT) thereof was observed using a cobalt filter.

5 μL of 2% NaFI (sodium fluorescein) containing no preservative agent was administered into eyes of the examinee using an eppendorf pipette. Immediately after the administration of NaFI, the head of the examinee was located at the headrest, and the examinee blinked his eyes three times and then stared to the front.

The tear film located on the cornea of the examinee was monitored by the slit lamp microscope, and simultaneously the time taken from the time that the examinee finally blinked his eyes until one or more black spots appeared on the tear film was measured.

The tear break up times (TBUTs) of the examinees was measured with respect to each eye, and the average values are given in Table 13 below.

[Table 13] Tear breakup times of examinees

(2) Measurement of tear amount

The tear amounts of examinees were measured using a Schirmer test.

A filter paper was placed at the bottom of an eyelid, and then the length of the filter paper wetted by tears was measured. The results thereof are given in Table 14 below.

[Table 14] Tear amounts of examinees measured using Schirmer test

(3) Evaluation of symptoms attributable to xerophthalmia

The symptoms caused by xerophthalmia, such as burning, foreign-body sensation, itching, redness, eye pain, photophobia, blurred vision and the like, were scored by 1 (nonsymptomatic) to 5 (very serious) according to the degree thereof, and then the average values thereof were calculated. The results thereof are given in Table 15.

[Table 15] Evaluation of symptoms attributable to xerophthalmia

(4) Evaluation of degree of corneal damage.

In order to evaluate the degree of corneal damage of examinees, the corneas of the examinees were stained, and the staining intensity thereof was measured.

An examinee sat with his head supported by a headrest. A slit lamp microscope was set at a magnifying power of 10, and was placed such that the angle of the microscope to illumination is 60˚. The slit lamp was adjusted such that the distance between slits is 3 mm, and the degree of corneal damage was evaluated using a cobalt filter.

5 μL of 2% NaFI (sodium fluorescein) containing no preservative agent was administered into the eyes of the examinee using an eppendorf pipette. Immediately after the administration of NaFI, the head of the examinee was placed on the headrest.

5 minutes after the administration of NaFI, as shown in FIG. 1, the cornea of the left eye and the cornea of the right eye were divided into five regions of C (central), T (temporal), N (nasal), S (superior) and I (inferior), respectively. The staining intensities of the five regions were scored by 0 (non-stained) to 3 (strongly stained) according to the criteria of Table 16 below, and then the average values were calculated. The results are given in Table 17.

[Table 16]

[Table 17] Evaluation of staining intensity of cornea

(5) Evaluation of conventional eye drops

In order to the evaluation of satisfaction with conventional eye drops, the satisfaction was scored from 0 (very poor) to 9 (very good) with respect to the following questions, and then averages were calculated. The results are given in Table 18.

[Table 18] Evaluation of satisfaction with conventional eye drops

2. Evaluation of effect of eye-drop composition on xerophthalmia

The eye-drop composition prepared in Example 1 was administered into the eyes of examinees, and then the degree to which various symptoms including xerophthalmia had improved was observed.

The eye-drop composition of Example 1 was administered into the eyes of the examinees by 1 ~ 2 drops four times per day for four weeks. Tear breakup time, tear amount, symptom attributable to xerophthalmia, degree of corneal damage and satisfaction with the eye drop composition were evaluated when the test started, one week post-administration of the eye-drop composition and four weeks post-administration thereof. The evaluation of the tear breakup time, the tear amount, the symptom attributable to xerophthalmia, the degree of corneal damage and the satisfaction with the eye drop composition was conducted in the same manner as above, and the results thereof are given in Tables 19 to 23 below.

Further, the percentages of the persons answering that they did not feel the symptoms caused by xerophthalmia, such as burning, foreign-body sensation, itching, redness, eye pain, photophobia and blurred vision, are shown in FIG. 2. In order to accurately evaluate the percentages, a significance test was carried out at a level of the p values shown in FIG. 2 using a generalized estimating equation.

[Table 19] Tear breakup time of examinees after the administration of the eye-drop composition of Example 1

In order to accurately evaluate the tear breakup time given in Table 19 above, a significance test was carried out at a level of the p values given in Table 19 by repeatedly analyzing measurements.

[Table 20] Tear amount of examinees after the administration of the eye-drop composition of Example 1

In order to accurately evaluate the tear amount given in Table 20 above, a significance test was carried out at a level of the p values given in Table 20 by repeatedly analyzing measurements.

[Table 21] Evaluation of symptoms of xerophthalmia after the administration of the eye-drop composition of Example 1

In order to accurately evaluate the scores assigned by examinees given in Table 21 above, a significance test was carried out at a level of the p values given in Table 21 by repeatedly analyzing measurements.

[Table 22] Evaluation of staining intensity of cornea after the administration of the eye-drop composition of Example 1

In order to accurately evaluate the staining intensity of the cornea given in Table 22 above, a significance test was carried out at a level of the p values given in Table 22 by repeatedly analyzing measurements.

[Table 23] Evaluation of degree of satisfaction after the administration of the eye-drop composition of Example 1

In order to accurately evaluate the scores assigned by examinees given in Table 23 above, a significance test was carried out at a level of the p values given in Table 23 by repeatedly analyzing measurements.

As given in Tables 19 to 23, it is confirmed that, when the eye-drop composition of Example 1 was administered into the eyes of the examinees, the tear breakup time and the tear amount were increased, there was remarkable improvement in the symptoms caused by xerophthalmia, such as burning, foreign-body sensation, itching, redness, eye pain, photophobia and blurred vision, and the staining intensity of the cornea was greatly decreased to remarkably improve the damage to the cornea. Further, it is confirmed that the eye-drop composition was fast acting, the effect of the eye-drop composition persisted for a long time, and the xerophthalmia was effectively treated, and that the administration frequency of eye-drops was remarkably decreased. Particularly, it is confirmed that, even when the eye-drop compositions of Examples were administered into the eyes of the examinees for only one week, the effects of treating xerophthalmia were very rapidly exhibited.

From the above Experimental Examples, it can be seen that the eye-drop compositions of the present invention can effectively treat xerophthalmia, and can effectively control the proliferation and growth of microorganisms for a predetermined amount of time, so that the eye-drop compositions of the present invention can be used as multi-dosed formulation over a long period of time.

Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

Claims

An eye-drop composition for preventing or treating ocular diseases, comprising:

hyaluronic acid or a salt thereof, a magnesium ion, a sodium ion, a potassium ion, a chloride ion and, an aminocaproate ion, an edetate ion and a tetraborate ion.
The eye-drop composition of claim 1, further comprising a phosphate ion.
An eye-drop composition for preventing or treating ocular diseases, comprising:

about 0.15 ~ about 0.2 wt% of hyaluronic acid or a salt thereof;

about 0.25 ~ about 0.55 mmol/L of a magnesium ion;

about 50 ~ about 80 mmol/L of a sodium ion;

about 13 ~ about 18 mmol/L of a potassium ion;

about 60 ~ about 75 mmol/L of a chloride ion; and

at least one selected from the group consisting of an aminocaproate ion, an edetate ion, a tetraborate ion, and a phosphate ion.
The eye-drop composition of claim 3, wherein the composition comprises about 8.5 mmol/L or less of the aminocaproate ion, about 3.5 mmol/L or less of an edetate ion, and about 1.5 mmol/L or less of the tetraborate ion.
The eye-drop composition of claim 4, further comprising about 5 mmol/L or less of the phosphate ion.
The eye-drop composition of claim 3, wherein the composition comprises:

about 0.8 ~ about 8.5 mmol/L of the aminocaproate ion;

about 0.5 ~ about 3.5 mmol/L of the edetate ion; and

about 0.15 ~ about 1.5 mmol/L of the tetraborate ion.
The eye-drop composition of claim 6, further comprising about 0.5 ~ about 5 mmol/L of the phosphate ion.
The eye-drop composition of claim 3, wherein the salt of hyaluronic acid is sodium hyaluronate.
The eye-drop composition of claim 3, wherein hyaluronic acid or the salt of hyaluronic acid has a weight-average molecular weight of about 800,000 ~ about 1,600,000.
The eye-drop composition of claim 3, wherein the ocular disease is xerophthalmia.
The eye-drop composition of claim 3, wherein the ocular disease is a corneal and conjunctival epithelial cell disorder.
The eye-drop composition of claim 3, wherein the composition is used as an artificial tear.
The eye-drop composition of claim 3, wherein the composition is used as a multi-does formulation.
The eye-drop composition of claim 3, wherein the composition inhibits proliferation and growth of microbes.
An eye-drop composition for preventing or treating ocular diseases, comprising:

about 0.15 ~ about 0.2 wt% of hyaluronic acid or a salt thereof;

about 0.25 ~ about 0.55 mmol/L of a magnesium ion;

about 50 ~ about 80 mmol/L of a sodium ion;

about 13 ~ about 18 mmol/L of a potassium ion;

about 60 ~ about 75 mmol/L of a chloride ion;

about 0.8 ~ about 8.5 mmol/L of an aminocaproate ion;

about 0.5 ~ about 3.5 mmol/L of an edetate ion; and

about 0.15 ~ about 1.5 mmol/L of a tetraborate ion.
The eye-drop composition of claim 15, further comprising about 0.5 ~ about 5 mmol/L of a phosphate ion.
An eye-drop composition for preventing or treating ocular diseases, comprising:

about 0.1 ~ about 0.3 wt% of sodium hyaluronate;

about 0.1 ~ about 0.6 mmol/L of a magnesium ion;

about 40 ~ about 90 mmol/L of a sodium ion;

about 10 ~ about 20 mmol/L of a potassium ion;

about 40 ~ about 90 mmol/L of a chloride ion;

about 0.5 ~ about 10 mmol/L of an aminocaproate ion;

about 0.3 ~ about 7 mmol/L of an edetate ion; and

about 0.1 ~ about 3 mmol/L of a tetraborate ion.
The eye-drop composition of claim 17, further comprising about 0.3 ~ about 7 mmol/L of a phosphate ion.
An eye-drop composition for preventing or treating ocular diseases, which is prepared by mixing sodium hyaluronate, magnesium chloride hydrate, sodium chloride, potassium chloride, borax, aminocaproic acid, sodium edetate hydrate and water.
The eye-drop composition of claim 19, wherein the composition is prepared by further adding and mixing sodium hydrogenphosphate hydrate.
The eye-drop composition of claim 20, wherein the composition is prepared by mixing about 0.1 ~ about 0.3 wt% sodium hyaluronate, about 0.08~about 0.2mg/mL of magnesium chloride hydrate, about 2.5~about 3.5mg/mL of sodium chloride, about 0.5~about 1.5mg/mL of potassium chloride, about 0.8~about 1.5mg/mL of sodium hydrogenphosphate hydrate, about 0.05~about 1mg/mL of borax, about 0.1~about 1.5mg/mL of aminocaproic acid, about 0.1~about 0.5mg/mL of sodium edetate hydrate and water.
A method of preventing or treating ocular diseases by administrating an eye-drop composition which comprises hyaluronic acid or a salt thereof, a magnesium ion, a sodium ion, a potassium ion, a chloride ion and, an aminocaproate ion, an edetate ion and a tetraborate ion.
The method of claim 22, wherein the composition comprises:

about 0.1 ~ about 0.3 wt% of hyaluronic acid or a salt thereof;

about 0.1 ~ about 0.6 mmol/L of the magnesium ion;

about 40 ~ about 90 mmol/L of the sodium ion;

about 10 ~ about 20 mmol/L of the potassium ion;

about 40 ~ about 90 mmol/L of the chloride ion;

about 0.5 ~ about 10 mmol/L of the aminocaproate ion;

about 0.3 ~ about 7 mmol/L of the edetate ion; and

about 0.1 ~ about 3 mmol/L of the tetraborate ion.
The method of claim 22, wherein the composition comprises:

about 0.1 ~ about 0.3 wt% of hyaluronic acid or the salt thereof;

about 0.1 ~ about 0.6 mmol/L of the magnesium ion;

about 40 ~ about 90 mmol/L of the sodium ion;

about 10 ~ about 20 mmol/L of the potassium ion;

about 40 ~ about 90 mmol/L of the chloride ion;

about 0.5 ~ about 10 mmol/L of the aminocaproate ion;

about 0.3 ~ about 7 mmol/L of the edetate ion;

about 0.1 ~ about 3 mmol/L of the tetraborate ion; and

about 0.3 ~ about 7 mmol/L of a phosphate ion.
The method of claim 22, wherein the composition comprises:

about 0.15 ~ about 0.2 wt% of hyaluronic acid or a salt thereof;

about 0.25 ~ about 0.55 mmol/L of the magnesium ion;

about 50 ~ about 80 mmol/L of the sodium ion;

about 13 ~ about 18 mmol/L of the potassium ion;

about 60 ~ about 75 mmol/L of the chloride ion;

about 0.8 ~ about 8.5 mmol/L of the aminocaproate ion;

about 0.5 ~ about 3.5 mmol/L of the edetate ion; and

about 0.15 ~ about 1.5 mmol/L of the tetraborate ion.
The method of claim 22, wherein the composition comprises:

about 0.15 ~ about 0.2 wt% of hyaluronic acid or a salt thereof;

about 0.25 ~ about 0.55 mmol/L of the magnesium ion;

about 50 ~ about 80 mmol/L of the sodium ion;

about 13 ~ about 18 mmol/L of the potassium ion;

about 60 ~ about 75 mmol/L of the chloride ion;

about 0.8 ~ about 8.5 mmol/L of the aminocaproate ion;

about 0.5 ~ about 3.5 mmol/L of the edetate ion;

about 0.15 ~ about 1.5 mmol/L of the tetraborate ion; and

about 0.5 ~ about 5 mmol/L of a phosphate ion.