WO2009157596A1

WO2009157596A1 - Ophthalmic composition for preventing and treating ocular diseases

Info

Publication number: WO2009157596A1
Application number: PCT/KR2008/003606
Authority: WO
Inventors: Soo Hyun Sung; Jai Jun Choung; Sae Kwang Ku
Original assignee: Dhp Korea Ltd., Co.
Priority date: 2008-06-24
Filing date: 2008-06-24
Publication date: 2009-12-30
Also published as: KR100827400B1

Abstract

The present invention relates to an ophthalmic composition for the prevention and treatment of ocular diseases, comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof as active ingredients. The ophthalmic composition shows greater synergistic effects on eye moisturizing and lubrication through improved viscoelasticity, osmosis and permeability as compared with the known ophthalmic compositions, thereby being useful for the prevention and treatment of ocular diseases such as corneal and conjunctival epithelial disorders.

Description

OPHTHALMIC COMPOSITION FOR PREVENTING AND TREATING OCULAR DISEASES

Technical Field

[1] The present invention relates to an ophthalmic composition for the prevention and treatment of ocular diseases. Background Art

[2] Natural tears are composed of three layers: a lipid, an aqueous, and a mucous layer.

Dry eyes may be caused by a lack of tears or any one component to provide constant moisture and lubrication. Dry eye syndrome is an ocular disease that is caused by a chronic lack of sufficient lubrication and moisture in the eye, and represents various abnormal states involving a disorder of the tear film due to a problem with the quantity and quality of tears moisturizing the eyes. Dry eye syndrome, also called Keratoconjunctivitis sicca, may wax and wane over the course of time. Occasionally, it causes eye irritation, persistent pain and foreign body sensation, and if left untreated, this condition can lead to ulcers or scars on the cornea, and eventually loss of vision. Dry eye syndrome is commonly associated with a systemic inflammatory process, and it is often related to general health conditions.

[3]

[4] Meanwhile, the known anionic polymers such as hyaluronic acid and carboxymethyl- cellulose have been generally employed in ophthalmic solutions, because of their moisturizing and lubrication effects, which contribute to the alleviation of eye irritation. Currently, ophthalmic solutions containing sodium hyaluronate as an active ingredient have been used as an agent for treating a corneal and conjunctival disorder associated with an endogenous disease such as Sjogren's syndrome, Stevens-Johnsons syndrome, or dry eye syndrome; and a corneal epithelial disorder accompanied with an exogenous disease in the case of post-operation, drug use, trauma, or use of hard contact lenses, whereas ophthalmic solutions containing sodium carboxymethylcellulose as an active ingredient have been only used for eye fatigue and dry eyes. Up to now, single formulations of sodium hyaluronate or sodium carboxymethylcellulose have been only developed as ophthalmic compositions and made commercially available, and there have been no studies on the efficacy and combination of two ingredients, concerning ophthalmic solutions containing both of the ingredients. Further, it is problematic that single formulations are effective in treating mild to moderate symptoms, but not severe dry eye syndrome.

[5] Disclosure of Invention

Technical Problem

[6] Accordingly, the present inventors have developed a preservative-free ophthalmic composition comprising hyaluronic acid as an active ingredient, and during a manufacturing process, they have considered a combination of various active ingredients to be used in ophthalmic compositions, so as to examine the efficacy according to combination ratio. As a result, they found that compared to each single formulation, an ophthalmic composition prepared by blending hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof in a predetermined ratio shows favorable effects on dry eye syndrome through combining a moisturizing effect due to high viscoelasticity of hyaluronic acid and a lubrication effect due to the surfactant property of carboxymethylcellulose, thereby completing the present invention. Technical Solution

[7] It is an object of the present invention to provide an ophthalmic composition for the prevention and treatment of ocular diseases with established safety and efficacy, which comprises hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof as active ingredients for the effective treatment of ocular diseases. Brief Description of Drawings

[8] FIG. 1 shows histopathological observations of the cornea (A and B; intact eye, C and D; dry eye, E and F; CMC-treated group, I and J; DHP- 101 -treated group, K and L; DHP-300-treated group, and M and N; DHP-500-treated group), in which ThT indicates the thickness of total cornea, ThE the thickness of epithelium, and ThS the thickness of stroma;

[9] FIG. 2 shows histopathological observations of the conjunctiva (A and B; intact eye,

C and D; dry eye, E and F; CMC-treated group, I and J; DHP- 101 -treated group, K and L; DHP-300-treated group, and M and N; DHP-500-treated group), in which ThE indicates the thickness of epithelium;

[10] FIG. 3 is the result of tear volume measurement by Schirmer test;

[11] FIG. 4 is the result of measuring corneal fluorescein permeability;

[12] FIG. 5 shows histopathological observations of the cornea (A and B; intact eye, C and D; dry eye, E and F; CMC-treated group, G and H; SH-treated group, I and J; DHP- 101 -treated group), in which ThT indicates the thickness of total cornea, ThE the thickness of epithelium, and ThS the thickness of stroma;

[13] FIG. 6 shows histopathological observations of the bulbar conjunctiva (A; intact eye,

B; dry eye, C; CMC-treated group, D; SH-treated group, and E; DHP- 101 -treated group);

[14] FIG. 7 shows histopathological observations of the palpebral conjunctiva (A; intact eye, B; dry eye, C; CMC-treated group, D; SH-treated group, and E; DHP- 101 -treated group);

[15] FIG. 8 shows Caspase-3-immunoreactivity observations of the corneal epithelium

(A; intact eye, B; dry eye, C; CMC-treated group, D; SH-treated group, and E; DHP- 101 -treated group); and

[16] FIG. 9 shows PARP-immunotreactivity observations of the corneal epithelium (A; intact eye, B; dry eye, C; CMC-treated group, D; SH-treated group, and E; DHP- 101-treated group). Best Mode for Carrying out the Invention

[17] In accordance with one aspect, the present invention relates to an ophthalmic composition for the prevention and treatment of ocular diseases, comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof as active ingredients.

[18]

[19] Hyaluronic acid (HA) of the present invention is a major component of extracellular matrix tissues, which is widely distributed in the stroma of the connective tissue so as to create a visco-elastic solution under physiological conditions, thereby providing mechanical protection and cushioning for tissues such as the iris and retina, and the corneal cells, vascular endothelial cells, and epithelium cells. In addition, it is known that hyaluronic acid is used as an ophthalmic surgical aid during surgical procedures on the vitreous body or the like, and sodium hyaluronate has been useful as an agent for treating a corneal epithelial disorder associated with an endogenous disease such as Sjogren's syndrome, Stevens-Johnsons syndrome, or dry eye syndrome; and a corneal epithelial disorder accompanied with an exogenous disease in the case of post- operation, drug use, trauma, or use of hard contact lenses. Tearin Free Eye Drops (manufactured in Korea) and VIVIAL Eye Drops (imported), which are prepared by containing 0.1% by weight of sodium hyaluronate as a single active ingredient, are commercially available.

[20] Further, carboxymethylcellulose (CMC) of the present invention is a cellulose derivative with carboxymethyl groups bound to some of the hydroxyl groups present in the glucopyranose monomers that form the cellulose backbone. Carboxymethylcellulose is generally used in food science as a viscosity modifier, because it has high viscosity and is non-toxic. In addition, its sodium salt is often used as a lubricant in artificial tears.

[21] However, as mentioned above, there is no current attempt to use a composition containing hyaluronic acid or a pharmaceutically acceptable salt thereof, and car- boxymethylcellulose or a pharmaceutically acceptable salt thereof for the prevention and treatment of ocular diseases. Specifically, it has not been considered that the mixture of hyaluronic acid and carboxymethylcellulose is applied to ocular diseases, since their application is different in accordance with specific types of ocular diseases and application time. Thus, the present inventors have confirmed that hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof shows greater synergistic effects when mixed with each other, compared to each single formulation.

[22]

[23] In the present invention, hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof may be prepared by chemical or enzymatic methods known in the art, or may be purchased from commercially available sources.

[24]

[25] As used herein, the term, 'Pharmaceutically acceptable salt' refers to salts of acidic or basic groups, which can be present in hyaluronic acid and carboxymethylcellulose of the present invention, unless otherwise specified. Examples of the pharmaceutically acceptable salt include sodium salts, potassium salts, and calcium salts, and preferably sodium salts.

[26]

[27] As used herein, the term 'Prevention and treatment of ocular disease' refers to the prevention and treatment of a corneal epithelial disorder associated with an endogenous disease such as Sjogren's syndrome, Stevens-Johnsons syndrome, or dry eye syndrome; and a corneal epithelial disorder accompanied with an exogenous disease in the case of post-operation, drug use, trauma, or use of hard contact lens. The ocular diseases preferably include, but are not limited to, dry eye syndrome.

[28]

[29] The composition for the prevention and treatment of ocular diseases of the present invention comprises hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof in a weight ratio of 1: 0.8 to 5.2, preferably 1: 0.8 to 3.2, more preferably 1: 1 to 2.5, and even more preferably 1: 2. In terms of pharmaceutics, if the weight ratio is less than the above range, the viscoelasticity is too low to provide sufficient moisture and lubrication, and if the weight ratio is more than the above range, there is a problem in permeability due to an excessively high viscoelasticity, so as not to ensure that ophthalmic compositions retain suitable rheological properties.

[30] Further, the composition according to the present invention was confirmed to have a favorable effect on the prevention and treatment of ocular diseases, compared to each single formulation, by histopathological observations of corneal damage, thicknesses of total cornea, epithelium and stroma, and changes in the damaged regions, and histopathological observations of focal desquamation of conjunctiva, mucous- producing cell numbers, and conjunctiva epithelial thickness according to each weight ratio. Therefore, the effects on eye fatigue and dry eyes, and efficacy of sodium hyaluronate-containing ophthalmic compositions, can be maximized by using the mixed formulation of hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof in a weight ratio of 1: 0.8 to 5.2, preferably 1: 0.8 to 3.2, more preferably 1: 1 to 2.5, and even more preferably 1 : 2.

[31]

[32] It is preferable that hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof of the present invention is contained in an amount of 0.1 to 50% by weight, based on the total weight of the composition of the present invention.

[33]

[34] The composition of the present invention may further comprise a pharmaceutically acceptable carrier, in addition to hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof. Examples of the pharmaceutically acceptable carrier may include isotonic agents, buffering agents, stabilizing agents, pH adjusting agents, and solvents.

[35]

[36] The isotonic agent functions to assure approximate physiologic tonicity to the ophthalmic composition, and may be one or more selected from sodium chloride and potassium chloride. In the present invention, the isotonic agent is preferably used in an amount of 150 to 400 parts by weight, based on 100 parts by weight of the active ingredients.

[37] The buffering agent is used to regulate acidity or alkalinity of the ophthalmic composition, so as to contact the eye without causing irritation, and it is therefore important that the ophthalmic composition is an isotonic solution having the same or similar pH to natural mammalian eye fluids. Typically, this requires that the pH of the ophthalmic composition be between 7.1 to 7.3. Accordingly, it is preferable that the buffering agent is used in an amount of 50 to 150 parts by weight, based on 100 parts by weight of the active ingredients. The buffering agent that is used to prepare the ophthalmic composition according to the present invention is preferably one or more selected from aminocaproic acid, dibasic sodium phosphate, and monobasic sodium phosphate. As a buffering agent used in the ophthalmic composition of the present invention, the phosphate concentration helps to prevent sight-threatening complications, and very high concentrations, namely, nonphysiological levels of phosphate (higher than 71.45 mmol/L) may favour the formation of insoluble crystalline calcium phosphate deposits when used on a damaged corneal surface.

[38] The stabilizing agent functions to stabilize the ophthalmic compositions, and is preferably used in an amount of 1 to 15 parts by weight, based on 100 parts by weight of the active ingredients. If the stabilizing agent is less than 1 part by weight, there is a problem in that the stability of the ophthalmic solution cannot be assured. If the stabilizing agent is more than 15 parts by weight, the stability of the ophthalmic solution may be also deteriorated. The stabilizing agent may be one or more selected from edetate sodium and sodium perborate.

[39] The pH adjusting agent functions to adjust natural tears to the physiological pH, and examples thereof may include one or more selected from hydrochloric acid and sodium hydroxide. In the present invention, the pH adjusting agent is used in a suitable amount to adjust the ophthalmic composition to the pH range of 7.1 to 7.3.

[40]

[41] The ophthalmic composition according to the present invention may be formulated into either preservative-containing or preservative-free ophthalmic solutions. Examples of the preservative used to prepare the preservative-containing ophthalmic compositions include one or more selected from benzalkonium chloride, methylparaben, and ethylparaben, and it is preferably used in an amount of 5 to 15 parts by weight, based on 100 parts by weight of the active ingredients. However, since the ophthalmic solution for dry eye is frequently used for a long period of time, the preservative may cause eye irritation, resulting in corneal damage. Therefore, it is preferable to use preservative-free ophthalmic solutions.

[42]

[43] In the case where the ophthalmic composition according to the present invention is formulated into a liquid form, the ophthalmic composition further comprises a solvent, and as the solvent, sterile purified water or injectable distilled water is preferably used. In this regard, it is necessary to determine the content of solvent according to the total volume of ophthalmic solution. That is, in addition to the ophthalmic composition, a solvent is contained in a suitable amount in the total ophthalmic solution. The ophthalmic composition according to the present invention is preferably in the form of liquid formulation.

[44] The ophthalmic composition according to the present invention is preferably added to the eye in an amount of 1 drop five to six times a day, but the dosage may vary depending on an individual's symptoms.

[45] [46] The ophthalmic composition according to the present invention comprises a mixed formula of hyaluronic acid having a moisturizing effect due to its high viscoelasticity and carboxymethylcellulose having a lubrication effect due to its surfactant property, thereby showing favorable synergistic effects for the prevention and treatment of ocular diseases, compared to each single formulation of hyaluronic acid and carboxymethylcellulose or salts thereof, which was confirmed by histopathological observations of corneal damage, thicknesses of total cornea, epithelium and stroma, and changes in the damaged regions, and histopathological observations of focal desquamation of conjunctiva, mucous -producing cell numbers, and conjunctiva epithelial thickness. Accordingly, the composition of the present invention is able to overcome drawbacks of the known single formulations, and achieve more favorable effects for the treatment of ocular diseases. Mode for the Invention

[47] Hereinafter, the present invention will be described in more detail with reference to the following Examples. However, it will be apparent to those skilled in the art that these Examples are for the illustrative purpose only and the invention is not intended to be limited by these Examples.

[48]

[49] Example 1: Preparation of preservative-free ophthalmic solution

[50] According to the contents of the following Table 1, the components were precisely weighed, and then sterile purified water was added as a solvent to a preparation flask equipped with a stirrer. Active ingredients, an isotonic agent, a buffering agent, and a stabilizing agent were also added thereto, and dissolved under stirring. A pH adjusting agent was added to the solution, so as to adjust the pH to 7.1. The prepared solution was sterilized, and filtered with a 0.2 μm filter. Then, the solution was filled into a small bottle, and sealed with a packing and cap to prepare an ophthalmic solution.

[51]

[52] Table 1

[Table 1] [Table ]

[53] [54] Example 2: Preparation of preservative-free ophthalmic solution [55] According to the contents of the following Table 2, the components were precisely weighed, and an ophthalmic solution was prepared in the same manner as in Example 1.

[56] Table 2

[Table 2] [Table ]

[57] [58] Example 3: Preparation of preservative-containing ophthalmic solution [59] According to the contents of the following Table 3, the components were precisely weighed, and then sterile purified water was added as a solvent to a preparation flask equipped with a stirrer. Active ingredients, an isotonic agent, a buffering agent, and a stabilizing agent were also added thereto, and dissolved under stirring. A preservative was added to the solution, and dissolved under stirring. Subsequently, a pH adjusting agent was added to the solution, so as to adjust the pH to 7.1. The prepared solution was sterilized, and filtered with a 0.2 μm filter. Then, the solution was filled into a small bottle, and sealed with a packing and cap to prepare an ophthalmic solution.

[60] [61] Table 3 [Table 3] [Table ]

[62] [63] Example 4: Preparation of preservative-containing ophthalmic solution [64] According to the contents of the following Table 4, the components were precisely weighed, and an ophthalmic solution was prepared in the same manner as in Example 3.

[65] [66] Table 4

[Table 4] [Table ]

[67] [68] Experimental Example 1: Effects of the composition of the present invention according to composition ratio

[69] The mixed formulations of sodium carboxymethylcellulose and sodium hyaluronate were prepared by varying the composition ratio (DHP-101: 0.1% sodium hyaluronate + 0.2% sodium carboxymethylcellulose; DHP-300: 0.1% sodium hyaluronate + 0.3% sodium carboxymethylcellulose; and DHP-500: 0.1% sodium hyaluronate + 0.5% sodium carboxymethylcellulose ), and their protective effects on the cornea and conjunctiva were examined by histopathological observations, compared to the single formulations (Table 5).

[70] Table 5 [Table 5] [Table ]

[71] [72] * DHP-101: 0.1% sodium hyaluronate + 0.2% sodium carboxymethylcellulose [73] * DHP-300: 0.1% sodium hyaluronate + 0.3% sodium carboxymethylcellulose [74] * DHP-500: 0.1% sodium hyaluronate + 0.5% sodium carboxymethylcellulose [75] * Dial Clear™: 0.5% sodium carboxymethylcellulose (CMC) [76] * Tearin Fresh™: 0.1% sodium hyaluronate (SH) [77] [78] 1-1. Induction of dye eye and treatment of test materials [79] The rats were anesthetized by 25 mg/kg intraperitoneal injection of Zoletile mixture (Zoletile 50; Virbac Lab., France). After deep anesthesia was achieved, the central region of the corneal epithelium (0.4mm²) was scraped mechanically with an ophthalmic surgical blade. Just after scraping, the rats were placed in a desiccation chamber with room temperature of 28 C, relative humidity of 25-30%, and constant air flow (2.4 m/sec), and maintained for 5 hours. The intact control group was not exposed to dry air-flow, after scraping the corneal epithelium.

[80] 5 jΛ /rat of test materials and reference materials were added dropwise to the eyes at 30 minutes before initiation of exposure to air- flow, and then every 30 min after initiation of exposure to air-flow (total 11 times) for 5 hours, followed by histopathological observation.

[81] [82] 1-2. Histopathological examination [83] The rats were killed and the eyeballs with bulbar conjunctiva were removed and fixed in Davidson's solution (37.5% ethanol, 12.5% acetic acid, and 25% formaldehyde [37% solution]). The corneal specimens were embedded in paraffin, cross-sectioned, and stained with HE (hematoxylin-eosin) for observing cornea and with PAS (periodic acid Schiff) for observing mucous -producing goblet cells in the cornea.

[84] Total thicknesses of cornea (βia), corneal epithelial thickness (βia), cornea stroma thickness (/M), damaged corneal epithelium percent region (per 5 mm of corneal epithelial lining), thickness of bulbar conjunctiva epithelium (jm), and number of goblet cells, goblet cell occupation percent region, and epithelial damage (calculated as percent damaged region among 1 mm of epithelial lining) were measured using a digital image analyzer (DMI-300, DMI, KOREA).

[85]

[86] 1-3. Histopathological changes of corneal epithelium

[87] The following Table 6 represents histomorphometrical changes detected in the rat cornea after 5 hrs exposure to dry air-flow (5 /M/eye, 30 min intervals).

[88]

[89] Table 6

[Table 6] [Table ]

[90] (*values are expressed as Mean SD of ten rats; CMC, sodium carboxymethyl- cellulose ; SH, sodium hyaluronate; ^ap<0.01 and ^bp,0.05 compared with intact group; ^c p<0.01 and ^dp<0.05 compared with dry eye control)

[91] [92] As compared with intact control, focal desquamation of corneal epithelial lining was detected in dry eye control group with significant (p<0.01) decreases of total thickness of cornea, corneas epithelial and stromal thicknesses, and significant (p<0.01) increases of epithelial damage, respectively.

[93] Corneal damage was significantly decreased in all of the drug-treated groups. As compared with dry eye control group, the total thickness of cornea was increased in order of DHP-101, DHP-300 and DHP-500 treated groups, and corneas epithelial thickness was increased in order of DHP-300, DHP-101 and DHP-500 treated groups; and epithelial damage was decreased in DHP-500, DHP-300 and DHP-101 treated groups, as compared with single formulations of sodium hyaluronate and sodium car- boxymethylcellulose (see Table 6 and FIG. 1).

[94] Specifically, the total corneal thickness was -24.40% in dry eye control, as compared with intact control, but +15.61%, +10.21%, +20.45%, +14.56% and +7.39% in sodium carboxymethylcellulose and sodium hyaluronate (single formulation)-treated groups, and DHP-101, DHP-300 and DHP-500 (mixed formulations)-treated groups, as compared with dry eye control, respectively.

[95] The corneal epithelial thickness was -30.10% in dry eye control, as compared with intact control, but +20.27%, +5.53%, +33.68%, +37.05% and +15.29% in sodium carboxymethylcellulose and sodium hyaluronate (single formulation)-treated groups, and DHP-101, DHP-300 and DHP-500 (mixed formulations)-treated groups, as compared with dry eye control, respectively.

[96] The corneal stroma thickness was -14.49% in dry eye control, as compared with intact control, but +1.80%, +6.83%, +7.59%, +7.18%, and +0.24% in sodium carboxymethylcellulose and sodium hyaluronate (single formulation)-treated groups, and DHP-101, DHP-300 and DHP-500 (mixed formulations)-treated groups, as compared with dry eye control, respectively.

[97] Epithelial damage was +382.19% in dry eye control, as compared with intact control, but -18.05%, -44.68%, -58.56%, -62.56% and -64.64% in sodium carboxymethylcellulose and sodium hyaluronate (single formulation)-treated groups, and DHP-101, DHP-300 and DHP-500 (mixed formulations)-treated groups, as compared with dry eye control, respectively.

[98] In accordance with histopathological observations of the corneal epithelium, it can be seen that DHP-101 and DHP-300 showed more favorable effects in all the histopathological observations than sodium carboxymethylcellulose and sodium hyaluronate (single formulation)-treated groups, and DHP-500 also showed more favorable effects on corneal epithelial damage than other formulations.

[99]

[100] 1-4. Histopathological changes of conjunctiva

[101] The following Table 7 represents histomorphometrical changes detected in the rat conjunctiva after 5 hrs exposure to dry air-flow (5 /M/eye, 30 min intervals).

[102] Table 7 [Table 7] [Table ]

[103] (*values are expressed as Mean SD of ten rats; CMC, sodium carboxymethyl- cellulose ; SH, sodium hyaluronate; ^ap<0.01 and ^bp,0.05 compared with intact group; ^c p<0.01 and ^dp<0.05 compared with dry eye control.)

[104] [105] As compared with intact control, focal desquamation of conjunctiva epithelial lining and decrease of mucous -producing cell number were detected in dry eye control group with significant (p<0.01) decreases of conjunctiva epithelial thickness, number and percent of mucous-producing cells, and significant (p<0.01) increases of the damaged epithelial regions, respectively. However, these histopathological changes on conjunctiva were significantly decreased in all of the drug-treated groups. As compared with dry eye control, the conjunctiva epithelial thicknesses were increased in order of sodium carboxymethylcellulose « sodium hyaluronate < DHP-101 < DHP-500 < DHP-300-treated groups; mucous-producing cell numbers were increased in order of sodium carboxymethylcellulose = sodium hyaluronate « DHP-101 < DHP-500 < DHP-300-treated groups. Further, mucous-producing cells percents were more increased in DHP-500, DHP-300 and DHP- 101 -treated groups than sodium hyaluronate- and sodium carboxymethylcellulose (single formulation) -treated groups. However, epithelial damage was more decreased in DHP-500, DHP-300 and DHP- 101 -treated groups than sodium hyaluronate- and sodium carboxymethylcellulose (single formulation)-treated groups (see Table 7 and FIG. 2).

[106] Specifically, the conjunctiva epithelial thickness was -43.51% in dry eye control as compared with intact control, but +20.39%, +38.34%, +51.19%, +71.75%, and +61.89% in sodium carboxymethylcellulose, sodium hyaluronate, DHP-101, DHP-300 and DHP-500-treated groups, as compared with dry eye control, respectively.

[107] The mucous-producing cell numbers in the conjunctiva epithelium were -78.63% in dry eye control as compared with intact control, but +108.00%, +108.00%, +184.00%, +244.00% and +228.00% in sodium carboxymethylcellulose, sodium hyaluronate, DHP-101, DHP-300 and DHP-500-treated groups, as compared with dry eye control, respectively.

[108] The mucous-producing cell percents in the conjunctiva epithelium were changed as - 80.37% in dry eye control as compared with intact control, but they were changed as 134.13, 174.57, 218.89, 225.41 and 227.37% in sodium carboxymethylcellulose , sodium hyaluronate, DHP-101, DHP-300 and DHP-500-treated groups, as compared with dry eye control, respectively.

[109] The damaged epithelial regions were changed as 822.24% in dry eye control as compared with intact control, but they were changed as -22.29, -55.73, -61.35, -67.29 and -71.24% in sodium carboxymethylcellulose , sodium hyaluronate, DHP-101, DHP- 300 and DHP-500-treated groups, as compared with dry eye control, respectively.

[110] In accordance with histopathological observations of the conjunctiva, it can be seen that DHP-101, DHP-300 and DHP-500 showed more favorable effects on conjunctiva epithelial thickness, number and occupation region of mucous-producing cells, and epithelial damage (%, per 1 mm of epithelial lining) than sodium carboxymethylcellulose and sodium hyaluronate (single formulation) -treated groups.

[I l l]

[112] Experimental Example 2: Safety test

[113] Upon completion of the acclimatization period, body weight was measured

(HW-60KGL, AND, Korea) and animals with no symptoms were examined using a direct ophthalmoscope (Part Number: 1127-P-1005, Keeler Instruments Inc.). 9 rabbits with both healthy eyes were selected, and divided into groups.

[114] In a non- washing group (6 rabbits), inferior palpebra of the right eye was pulled from the eyeball to form a cup shape, and 0.1 mL of test material (ophthalmic solution of Example 1, hereinbelow referred to as DHP-101) was dropped into the conjunctival sac. The lids were then held together for about 1 sec to prevent loss of the test material, and the left eye was left untreated as a control.

[115] In a washing group (3 rabbits), the same amount of test material was dropped into the conjunctival sac of the right eye. After 30 sec, the eye was washed with 20 mL of saline solution (Lot No.: AAX7CV, Choongwae Pharma Corp., Korea) for about 30 sec without irritation. The left eye was also washed as a control.

[116] [117] 2-1. Observation of general symptoms and weight measurement [118] During the observation period, animal health conditions were observed once a day. On day 0 and 7 after treatment, body weights were measured using an electronic scale (HW-60KGL, AND, Korea).

[119] As a result, during the observation period, no animal death was observed, and changes in general symptoms, caused by treatment of the test material, were not observed (see Table 8).

[120] Further, the body weight of all animals increased after treatment of the test material. The non-washing group had a mean weight gain of 0.09 kg, and the washing group had a mean weight gain of 0.11 kg (see Table 9).

[121] [122] Table 8 [Table 8] [Table ]

[123] Table 9

[Table 9] [Table ]

[124] [125] 2-2. Observation of ocular lesions [126] As a control, the left eye was not treated with test materials, and on day 1, 2, 3, 4 and 7 after treatment with test materials, changes in the degree of corneal opacity, the opalescent area of the cornea, iris response, conjunctival redness, and conjunctival edema and secretions were observed with the naked eye. Ocular lesions were scored according to 'Draize' scale for scoring of ocular lesions'. During this period, the treated regions were photographed at each observation time.

[127] <Draize's scale for scoring of ocular lesions> [128]

1. Cornea

A Opacity-degree of density (area most dense taken for reading)

No Opacity 0

Scattered or diffuse area, details of iris clearly visible 1"

Easily discernible translucent areas, details of iris slightly obscured 2^*

Opalescent areas, no details of iris visible, size of pupil barely discernible 3^*

Opaque, iris invisible 4^*

B. Area of cornea involved

One quarter (or less) but not zero ]

Greater than one quarter, but less than half. 2

Greater than half, but less than three quarters 3

Greater than three quarters, up to whole area 4

A X B X 5 Total Maximum = 80

A Values

Normal 0

Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive) l'

No reaction to light, hemorrhage, gross destruction (any or all of these) 2^*

A X 5 Total Maximum = 10

3. Conjunctivae

A Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)

Vessels normal 0

Vessels definitely injected above normal 1

More diffuse, deeper crimson red, individual vessels not easily discernible 2

Diffuse beefy red 3

B. Chemosis

No swelling 0

Any swelling above normal (includes nictitating membrane) 1

Obvious swelling with partial eversion of lids 2

Swelling with lids about half-closed 3

Swelling with lids about half-closed to completely closed 4

C. Discharge

No discharge 0

Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1

Discharge with moistening of the lids and hairs just adjacent to lids 2

Discharge with moistening of the lids and hairs, and considerable area around the eye 3

Score (A + B + C) X 2 Total Maximum = 20

Total Maximum Score: 110 represents the sum of all scores obtained for the cornea, iris and conjunctivae.

[129]

[130] In the non-washing group, ocular lesions such as corneal opacity, iris response, conjunctival redness, and conjunctival edema and secretions were not observed on day 1, 2, 3, 4 and 7 after treatment with test materials, and Individual Total Score (I.T.S.) was divided by the number of animal to get the Mean Index of Ocular Irritation (M.I.O.I.) of '0'. In the washing group, ocular lesions such as corneal opacity, iris response, conjunctival redness, and conjunctival edema and secretions were not observed on day 1, 2, 3, 4 and 7 after treatment with test materials, and each Mean Index of Ocular Irritation (M.I.O.I.) was '0'.

[131]

[132] 2-3. Evaluation of ocular irritation

[133] The scale for scoring of ocular lesions was used to calculate corneal lesions (maximum value 80 points), iris lesions (maximum value 10 points) and conjunctival lesions (maximum value 20 points) at each observation time, and then the total score was calculated for each animal. Individual Total Score (LT. S.) was divided by the number of animals to obtain the Mean Index of Ocular Irritation (M.I.O.I.). Ocular irritancy was evaluated by LA. O.I. (Index of Acute Ocular Irritation) that is the maximum value of M.I.O.I. and Day 7 I.I.O.I. (Individual Index of Ocular Irritation) with reference to Guillot's ocular irritation score index, as follows. The results are shown in the following Table 10.

[134]

[135] Note: The table above is impossible to understand. I have no idea what the data is, what it means, how it is organized, what the units are, etc. It's basically useless.

[136] [137] Table 10 [Table 10] [Table ]

[138] [139] As shown in Table 10, when ocular irritancy of the test material DHP-101 was evaluated with reference to Guillot's ocular irritation score index, both I. A.O.I, and M.I.O.I. at 48 hours after treatment were evaluated as '0', thereby being determined as a nonirritant material.

[140] [141] Experimental Example 3: Pharmacological efficacy test of the composition of the present invention

[142] Among the mixed formulations of sodium carboxymethylcellulose and sodium hyaluronate, the efficacy of DHP-101 (0.1% sodium hyaluronate + 0.2% sodium carboxymethylcellulose ) as measured by changes in the tear volumes and corneal permeability, histopathological changes in the cornea and conjunctiva, and changes in caspase-3 immunoreactivity and PARP immunoreactivity were examined by comparison with the single formulations, so as to confirm the protective effects of the composition according to the present invention.

[143] [144] 3-1. Induction of dry eve and treatment of test material [145] The rats were anesthetized by 25 mg/kg intraperitoneal injection of Zoletile mixture (Zoletile 50; Virbac Lab., France). After deep anesthesia was achieved, the central region of the corneal epithelium (0.4 mm²) was scraped mechanically with an ophthalmic surgical blade. Just after scraping, the rats were placed in a desiccation room with room temperature of 28 C, relative humidity of 25-30%, and constant air flow (2.4 m/sec), and maintained for 5 hours. An intact control group was not exposed to dry air-flow after scraping the corneal epithelium.

[146] 5 /i6/rat of test material and reference materials (see the following Table 11) was added dropwise to the eye at 30 minutes before initiation of exposure to air-flow, and then every 1 hour after initiation of exposure to air- flow (total 6 times) for 5 hours, followed by observation.

[147] [148] Table 11 [Table 11] [Table ]

[149] *DHP-101:0.1% sodium hyaluronate + 0.2% sodium carboxymethylcellulose [150] [151] 3-2. Changes in tear volume (Schimer test) [152] The Schimer test using phenol red thread has been regarded as a routine method for detection of tear volumes and has been usually selected to test dry eye conditions (Fujihara et. al., 2001; Nakamura et al., 2004).

[153] Under anesthesia, 1x15 mm cobalt chloride paper (Toyo Rochi Kaisha, Japan) was placed on the temporal side of the lower eyelid margin for 60 seconds before initiation of exposure to dry air. The length of the moistened area from the edge was measured to an accuracy of 0.5 mm using an electronic digital caliper (Mytutoyo, Japan). Further, at the end of 5 hours dry air exposure and treatment of test material, the eyes were cleaned to remove extra- test material, and maintained for 60 seconds. Then, the Schimer test was performed in the same manner as before dry air exposure, so as to calculate the changes of tear volumes between before and after dry air exposure.

[154] [155] Although quite similar tear volumes were detected in all rats before dry air exposure, significant (p<0.01) decreases of tear volumes were detected after dry air exposure in dye eye control, and consequently the changes of tear volumes between before and after dry air exposure were significantly (p<0.01) decreased. However, these decreases in tear volumes were markedly inhibited in all of the drug-treated groups, especially DHP-101 showed significantly increases as compared with dry eye control (see Table 12 and FIG. 3).

[156] Table 12 [Table 12] [Table ]

[157] (*values are expressed as Mean SD of ten rats; CMC, sodium carboxymethyl- cellulose ; SH, sodium hyaluronate; ^ap<0.01 and ^bp,0.05 compared with intact group; ^c p<0.01 and ^dp<0.05 compared with dry eye control.)

[158] [159] The tear volumes before dry air exposure were +3.32% in a dry eye control as compared with an intact control, and +5.48%, -1.21% and +4.95% in CMC, SH, and DHP-101 treated groups as compared with dry eye control, respectively. The tear volumes after dry air exposure were -31.61% in a dry eye control as compared with an intact control, and +20.70%, +23.29% and +32.35% in CMC, SH, and DHP-101 treated groups as compared with dry eye control, respectively.

[160] The changes of tear volumes between before and after dry air exposure were - 628.23% in a dry eye control as compared with an intact control, but +31.14%, +39.00% and +80.78% in CMC, SH, and DHP 101 treated groups as compared with a dry eye control, respectively.

[161] [162] 3-3. Changes in corneal permeability [163] One of the most prevalent methods, fluorescent dye test has been used to detect the permeability of corneas. It is known that increased permeability correlates with corneal damages (Yokoori and Kinoshita, 1995; Nakamura et al., 2003, 2005; Steinfeld et al., 2004). Therefore, the damaged region can be easily detected using fluorescein. [164] In this Experimental Example, significant (p<0.01) increases of fluorescein intensity, indicating corneal permeability, were detected in dry eye control as compared with intact control. However, these increases of fluorescein intensity were significantly (p<0.01) inhibited in all of the drug-treated groups.

[165] The fluorescein intensity was 61.87% in a dry eye control as compared with an intact control, but -24.13%, -31.39% and -40.83% in CMC, SH and DHP 101 -treated groups as compared with a dry eye control, respectively (see Table 12 and FIG. 4).

[166]

[167] 3-4. Histopathological changes of corneal epithelium

[168] The rats were sacrificed, and the eyeballs with bulbar conjunctivas were removed and fixed in Davidson's solution (37.5% ethanol, 12.5% acetic acid and 25% formaldehyde [37% solution]), and palpebral conjunctivas were also removed and fixed in neutral buffered formalin. The eye and palpebral specimens were embedded in paraffin, cross- sectioned, and stained with HE (hematoxylin-eosin) for observing the cornea, and with PAS (periodic acid Schiff) for observing mucous-producing goblet cells of the cornea.

[169] Total thickness of cornea (/M), corneal epithelial thickness (/M), cornea stromal thickness (jm), corneal epithelial damage (%, per 5 mm of corneal epithelial lining), thickness of conjunctiva epithelium (/M), number and occupation percent region of mucous -producing cells and damaged epithelium percent region (per 1 mm of epithelial lining) were measured using a digital image analyzer (DMI-300, DMI, KOREA).

[170] Focal desquamation of cornea epithelial lining was detected in a dry eye control as compared with an intact control and consequently significant (p<0.01) decreases of total thickness of cornea, corneal epithelial and stromal thicknesses, and significant (p<0.01) increases of epithelial damage were also detected, respectively. However, these histopathological changes of the cornea were inhibited in all of the drug-treated groups, and especially the decreases of corneal epithelium thickness and increases of epithelial damage scores were more favorably inhibited by treatment of DHP-101 as compared with CMC and SH (single formulation)-treated groups.

[171] The total corneal thickness was -32.29% in a dry eye control as compared with an intact control, but +18.59%, +25.91% and +52.16% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively (see Table 13 and FIG. 5).

[172] Table 13 [Table 13] [Table ]

[173] (*values are expressed as Mean SD of ten rats; CMC, sodium carboxymethyl- cellulose; SH, sodium hyaluronate; ^ap<0.01 and ^bp,0.05 compared with intact group; ^c p<0.01 and ^dp<0.05 compared with dry eye control.)

[174] [175] Specifically, the corneal epithelial thicknesses were -42.66% in a dry eye control as compared with an intact control, but +29.84%, +37.31% and +52.16% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[176] The corneal stroma thicknesses were changed as -32.52% in dry eye control as compared with intact control, but they were changed as 19.49, 36.23 and 25.45% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[177] The damaged epithelial regions were changed as 901.32% in dry eye control as compared with intact control, but they were changed as -24.77, -38.85 and -64.70% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[178] 3-5. Histopathological changes in bulbar conjunctiva [179] Compared with intact control, focal desquamation of bulbar conjunctiva epithelial lining and decrease of mucous -producing cell number were detected in a dry eye control with significant (p<0.01) decreases of conjunctiva epithelial thicknesses, number and percent of mucous-producing cells, and significant (p<0.01) increases of the epithelial damage, respectively. However, these histopathological changes on bulbar conjunctiva were significantly inhibited in all of the drug-treated groups, and especially the changes were more favorably inhibited by treatment of DHP-101 as compared with CMC and SH (single formulation)-treated groups.

[180] Specifically, the bulbar conjunctiva epithelial thicknesses were changed as -45.59% in dry eye control as compared with intact control, but they were changed as 26.33, 41.38 and 74.61% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[181] The mucous-producing cell numbers in the bulbar conjunctiva epithelium were changed as -79.22% in dry eye control as compared with intact control, but they were changed as 168.75, 193.75 and 337.50% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[182] The mucous-producing cell regions in the bulbar conjunctiva epithelium were changed as -84.15% in dry eye control as compared with intact control, but they were changed as 298.33, 325.30 and 459.22% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[183] The damaged bulbar epithelial regions were changed as 739.52% in dry eye control as compared with intact control, but they were changed as -11.15, -46.13 and -71.35% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively (see Table 14 and FIG. 6).

[184]

[185] Table 14

[Table 14] [Table ]

[186] (*values are expressed as Mean SD of ten rats; CMC, sodium carboxymethyl- cellulose ; SH, sodium hyaluronate; ^ap<0.01 and ^bp,0.05 compared with intact group; ^c p<0.01 and ^dp<0.05 compared with dry eye control.)

[187] [188] 3-6. Histopathological changes in palpebral conjunctiva [189] Compared with intact control, focal desquamation of palpebral conjunctiva epithelial lining and decrease of mucous -producing cell number were detected in a dry eye control with significant (p<0.01) decreases of conjunctiva epithelial thicknesses, number and percent of mucous-producing cells, and significant (p<0.01) increases of epithelial damage, respectively. However, these histopathological changes on palpebral conjunctiva were significantly inhibited in all of the drug-treated groups, except for damaged epithelial regions of CMC treated group, in which similar damage scores were detected as compared with dry eye control. Especially, the changes were more favorably inhibited by treatment of DHP-101 as compared with CMC and SH-treated groups.

[190] Specifically, the palpebral conjunctiva epithelial thicknesses were changed as - 45.57% in dry eye control as compared with intact control, but they were changed as 26.01, 39.79 and 75.11% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[191] The mucous-producing cell numbers in the palpebral conjunctiva epithelium were changed as -71.97% in dry eye control as compared with intact control, but they were changed as 64.18, 123.88 and 232.84% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[192] The mucous-producing cell regions in the palpebral conjunctiva epithelium were changed as -65.71% in dry eye control as compared with intact control, but they were changed as 43.35, 84.46 and 137.50% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively.

[193] The damaged palpebral epithelial regions were changed as 1075.87% in dry eye control as compared with intact control, but they were changed as -2.45, -45.41 and - 65.91% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively (see Table 15 and FIG. 7).

[194] [195] Table 15 [Table 15] [Table ]

[196] (*values are expressed as Mean SD of ten rats; CMC, sodium carboxymethyl- cellulose ; SH, sodium hyaluronate; ^ap<0.01 and ^bp,0.05 compared with intact group; p<0.01 and ^dp<0.05 compared with dry eye control.)

[197] 3-7. Changes of Caspase-3 and PARP immunoreactivity [198] Caspase-3 and PARP are apoptotic mediators (Nunez et al., 1998; Barrett et al., 2001), and detection of the activated caspase-3 and PARP in the corneal epithelium indicates damage of the corneal epithelium by apoptosis, suggesting that these apoptotic mediators may be involved in the pathogenesis of dry eye (Yeh et al., 2003).

[199] [200] A significant (p<0.01) increase of caspase-3 immunoreactive regions was detected in a dry eye control as compared with an intact control. However, in all of the drug- treated groups, the caspase-3 immunoreactive regions were significantly (p<0.01) decreased in order of DHP-101 » SH > CMC as compared with dry eye control.

[201] The caspase-3 immunoreactive regions in the corneal epithelial lining were changed as 468.85% in dry eye control as compared with intact control, but they were changed as -25.61, -39.60 and -64.61% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively (Table 16 and FIG. 8).

[202] [203] Table 16 [Table 16] [Table ]

[204] (*values are expressed as Mean SD of ten rats; CMC, sodium carboxymethyl- cellulose ; SH, sodium hyaluronate; ^ap<0.01 and ^bp,0.05 compared with intact group; ^c p<0.01 and ^dp<0.05 compared with dry eye control.)

[205] A significant (p<0.01) increase of PARP immunoreactive regions was detected in dry eye control as compared with intact control. However, in all of the drug-treated groups, the PARP immunoreactive regions was significantly (p<0.01) decreased in order of DHP-101 » SH > CMC as compared with dry eye control.

[206] The PARP immunoreactive regions in the corneal epithelial lining were changed as 506.36% in dry eye control as compared with intact control, but they were changed as - 34.99, -44.45 and -65.20% in CMC, SH and DHP-101 treated groups as compared with dry eye control, respectively (Table 16 and FIG. 9). [207]

Industrial Applicability

[208] The ophthalmic composition according to the present invention is prepared by blending hyaluronic acid or a pharmaceutically acceptable salt thereof, and car- boxymethylcellulose or a pharmaceutically acceptable salt thereof as active ingredients in a predetermined ratio, thereby showing greater synergistic effects on eye moisturizing and lubrication through improved viscoelasticity, osmosis and permeability as compared with the known single formulations. Thus, the ophthalmic composition according to the present invention has better moisturizing and lubrication effects than single formulations of sodium hyaluronate and sodium carboxymethylcellulose, thereby being useful for the treatment and prevention of corneal epithelial disorders associated with endogenous diseases such as Sjogren s syndrome, Stevens-Johnsons syndrome, or dry eye syndrome; and corneal epithelial disorders accompanied with an exogenous diseases in the case of post-operation, drug use, trauma, or use of hard contact lenses, as well as eye fatigue and eye dry conditions.

[209]

Claims

[I] An ophthalmic composition for the prevention and treatment of ocular diseases, comprising hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof.

[2] The ophthalmic composition according to claim 1, wherein hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof is contained in a weight ratio of 1: 0.8 to 5.2.

[3] The ophthalmic composition according to claim 1, wherein the pharmaceutically acceptable salt is one selected from the group consisting of sodium, potassium, and calcium salts.

[4] The ophthalmic composition according to claim 1, wherein the composition further comprises one or more selected from an isotonic agent, a buffering agent, a stabilizing agent, and a preservative.

[5] The ophthalmic composition according to claim 4, wherein the isotonic agent is one or more selected from sodium chloride and potassium chloride.

[6] The ophthalmic composition according to claim 4, wherein the isotonic agent is contained in an amount of 150 to 400 parts by weight, based on 100 parts by weight of the active ingredients, hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof.

[7] The ophthalmic composition according to claim 4, wherein the buffering agent is one or more selected from aminocaproic acid, dibasic sodium phosphate, and monobasic sodium phosphate.

[8] The ophthalmic composition according to claim 4, wherein the buffering agent is contained in an amount of 50 to 150 parts by weight, based on 100 parts by weight of the active ingredients, hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof.

[9] The ophthalmic composition according to claim 4, wherein the stabilizing agent is one or more selected from edetate sodium and sodium perborate.

[10] The ophthalmic composition according to claim 4, wherein the stabilizing agent is contained in an amount of 1 to 15 parts by weight, based on 100 parts by weight of the active ingredients, hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof.

[I I] The ophthalmic composition according to claim 4, wherein the preservative is one or more selected from benzalkonium chloride, methylparaben, and ethylparaben.

[12] The ophthalmic composition according to claim 4, wherein the preservative is contained in an amount of 5 to 15 parts by weight, based on 100 parts by weight of the active ingredients, hyaluronic acid or a pharmaceutically acceptable salt thereof, and carboxymethylcellulose or a pharmaceutically acceptable salt thereof.

[13] The ophthalmic composition according to any one of claims 1 to 12, wherein the ophthalmic composition is formulated in a liquid form.

[14] The ophthalmic composition according to claim 1, wherein the ocular diseases are corneal and conjunctival epithelial disorders associated with endogenous diseases such as Sjogren s syndrome, Stevens-Johnsons syndrome, or dry eye syndrome; and corneal epithelial disorders accompanied with exogenous diseases in the case of post-operation, drug use, trauma.

[15] The ophthalmic composition according to claim 14, wherein the corneal and conjunctival epithelial disorder is dry eye syndrome.