KR20120107742A - Composition having effect on prevention and treatment of dry eye syndrome - Google Patents

Abstract

PURPOSE: A composition containing chitin-derived carboxymethyl chitosan is provided to suppress inflammation and apoptosis after xeroma and to promote goblet cell regeneration. CONSTITUTION: A composition for preventing and treating xeroma contaisn carboxymethyl chitosan(CMC) as an active ingredient. The CMC is a polysaccharide derived from chitin. The composition enhances the thickness of the conjunctiva and cornea and reduces the number of inflammatory cells. A pharmaceutical composition for preventing and treating xeroma contains the composition containing CMC. A composition is a topical application formulation such as eye drops, ointment, gel, or lotion.

Description

Composition having effect on prevention and treatment of Dry Eye Syndrome}

The present invention relates to a composition for preventing and treating dry eye syndrome, and more particularly, to a composition for preventing and treating dry eye syndrome containing carboxymethyl chitosan derived from chitin as an active ingredient.

Dry eye syndrome is a commonly encountered ocular surface disease. In the United States, about 5 million people over the age of 50 are estimated to have severe dry eyes [Schaumberg et al., Prevalence of dry eye syndrome among US women. Am J Ophthalmol 2003; 136 (2): 318-326. It is caused by damage to the ocular surface between the upper and lower eyelids and is associated with symptoms of eye discomfort. The National Eye Institute (NEI) / Industry Workshop classified dry eye conditions into two categories: aqueous deficient and evaporative (lipid deficient) [Lemp MA., Report of the National Eye Institute / Industry workshop on clinical trials in dry eyes. CLAO J 1995; 21: 221232. The tear film of the eye is thought to be composed of three layers: the innermost protein layer, the intermediate aqueous layer, and the outer lipid layer [Holly FJ and Lemp MA., Tear]. physiology and dry eyes. Surv Ophthalmol . 1977; 22: 6987], in fact, changes in the constituents within the tear film can lead to dry eye syndrome that causes the cornea to dry out. Dry eye syndrome can be used to avoid exacerbating factors, eyelid hygiene, tear supplementation, secretagogues, punctual plugs, anti-inflammatory agents, Non-pharmacologic and pharmacologic treatments, including environmental management such as moisture chambers and salivary gland autotransplantation for dry eye patients, can be administered [Behrens et al. , Dysfunctional tear syndrome study group. Dysfunctional tear syndrome: a Delphi approach to treatment recommendations. Cornea 2006; 25 (8): 900-907. Despite the many available items above, there is currently no ideal treatment for dry eye syndrome.

On the other hand, much effort and resources have been spent to develop reliable experimental models for severe dry eye or keratoconjunctivitis sicca (KCS). Thus, although some success has been achieved with animal models, no stable chronic models of human conditions have been reported [Barabino S and Dana MR., Animal models of dry eye: a critical assessment of opportunities and limitations. Invest Ophthalmol Vis Sci 2004; 45: 16411646, Dursun et al., A mouse model of keratoconjunctivitis sicca. Invest Ophthalmol Vis Sci 2002; 43: 632638. The currently available models also have problems such as short duration of disease, related systemic side effects, difficulty in surgery and incomplete development of KCS [Suwan-apichon et al., Botulinum toxin B-induced mouse model of keratoconjunctivitis sicca. Invest Ophthalmol Vis Sci 2006; 47 (1): 133-9].

Preservatives are an important component of ophthalmic preparations, providing antimicrobial activity and protecting the degradation of active drugs. However, the cytotoxic effect of the preservatives is prominent with prolonged use. The most common preservative in ophthalmic solvents for glaucoma and ocular surface diseases is benzalkonium chloride (BAC), which is used at a concentration of 0.01% (range, 0.004 to 0.02%) in topical multidose solutions. BAC is a quaternary ammonium compound and is known to accelerate the drying of the tear film, exacerbation of existing dry eye, and invasion of the cornea and conjunctiva. Epidemiologic data also showed that dry eye symptoms and signs were more prevalent when BAC-preserved drugs were used [Xiong et al., A rabbit dry eye model induced by topical medication of a preservative benzalkonium chloride. Invest Ophthalmol Vis Sci 2008; 49 (5): 1850-6].

Carboxymethyl chitosan (CMC) is a polysaccharide derived from chitin and is glycosaminoglycan, a major component of the corneal stromal extracellular matrix. Structurally similar to). CMC exhibits many advantageous biological properties such as bioadhesion, low toxicity, antioxidant and good biocompatibility. It also has physicochemical properties, which can make it a unique material for the manufacture of ocular drug delivery mediators. Ophthalmic preparations based on CMC significantly increased the shelf life of the drug prior to the cornea and showed antimicrobial and antifungal effects even when topically applying CMC at low concentrations [Hongyok et al., Effect of chitosan-N]. -acetylcysteine conjugate in a mouse model of botulinum toxin B-induced dry eye. Arch Ophthalmol 2009; 127 (4): 525-32].

Accordingly, the present inventors have made diligent research efforts to overcome the problems of the prior arts. As a result, when topically administering a composition containing carboxymethyl chitosan (CMC) to the dry eye animal model, the epithelial cell thickness of the cornea and conjunctiva is increased. To reduce the number of cells positive for inflammatory and apoptosis responses, it was confirmed that the tear amount can be restored to the normal range, the present invention was completed.

Therefore, the main object of the present invention is to provide a composition for treating and preventing dry eye syndrome containing carboxymethyl chitosan (CMC) as an active ingredient having a dry eye preventive and therapeutic effect.

Another object of the present invention to provide a pharmaceutical composition for treating and preventing dry eye syndrome containing the composition.

According to one aspect of the present invention, the present invention provides a composition for treating and preventing dry eye syndrome containing carboxymethyl chitosan (CMC) as an active ingredient.

The term “dry eye” (dry eye) is a disorder of the tear film that causes damage (including inflammation) to the ocular surface of the interpalpebral by the reduction or lack of tears or excessive evaporation. The definition and classification of dry eye disease: report of the Definition and Classification Subcommittee of the International Dry Eye WorkShop (2007). Ocul Surf . 2007 Apr; 5 (2): 75-92. Review].

In the composition of the present invention, the carboxymethyl chitosan (CMC) is characterized in that the polysaccharide (polysaccharide) derived from chitin (chitin).

The CMC is structurally similar to glycosaminoglycan, which is a major component of the extracellular matrix, and has bioadhesion, low toxicity, antioxidant, and good biocompatibility. Many advantageous biological properties, such as), can be used as a material for ocular drug delivery mediators.

The composition containing the CMC of the present invention as an active ingredient is characterized by increasing the epithelial thickness of the cornea and conjunctiva, and reducing the number of inflammatory cells.

In a specific embodiment of the present invention, the topical administration of the CMC composition of the present invention to the test animal causing dry eye was confirmed to significantly increase the epithelial thickness of the cornea and conjunctiva, the number of inflammatory cells was reduced compared to the control group. It confirmed that it became. In addition, the effect of the CMC composition was confirmed to be more effective than the commercially available hyalic acid (HA) solution (see Example 2 and FIGS. 4 and 5).

The composition of the present invention is characterized by reducing the number of caspase-3- and PARP positive cells involved in apoptosis in corneal and conjunctival epithelial cells and reducing the number of inflammatory cytokines, TNF-α positive cells in corneal stroma and conjunctival mucosa. It is done.

Through specific examples, the CMC composition according to the present invention reduces the number of caspase-3 and PARP positive cells of corneal and conjunctival epithelial cells in dry eye induced test animals, and also TNF-α positive in corneal stroma and conjunctival mucosa. It was confirmed that the number of cells was also reduced (see Example 2 and FIGS. 6 to 8).

An increase in caspase-3 expression and PARP expression is observed among the changes that occur in the process of apoptosis caused by cell damage. Therefore, it can be seen that apoptosis occurs when BAC is treated in the conjunctiva, and the number of caspase-3 and PARP positive cells that appear in one part of the apoptosis process is decreased when the CMC composition of the present invention is treated. Confirmed. Therefore, it can be seen that the CMC composition of the present invention has a protective effect or a reducing effect on apoptosis that appears upon cell damage. In addition, TNF-α immunostaining, a representative inflammatory cytokine, was performed to determine whether inflammation occurred when BAC was applied to the conjunctiva, and TNF-α-positive cell counts were observed when dry eye syndrome was induced by BAC treatment. And the number was reduced when treating the CMC composition of the present invention. Therefore, it can be seen that the CMC composition of the present invention reduces tissue damage or induced inflammatory response by BAC.

The "corneal epithelium" refers to a tissue consisting of 5 to 6 cell layers that constitute the top layer of the cornea. The cells constituting the corneal epithelium are detached 5-7 days after the base cell is formed from the stem cells of the periphery and moves to the center.

The "conjunctiva epithelium" is a cell constituting the uppermost layer of the conjunctiva, the goblet cells secreting mucin (mucin) is present in the conjunctival epithelium. The mucous component secreted from the goblet cells in the conjunctival epithelium constitutes a protein layer of the three components of the tear layer to play an important role in preventing the cornea from drying out.

As described above, it is known that dry eye can be treated if tear secretion is normally increased and cornea and conjunctiva function normally. [Nakamura et al., Protective effect of D-beta-hydroxybutyrate on corneal epithelia in dry eye conditions through suppression of apoptosis. Invest Ophthalmol Vis Sci . 2003 Nov; 44 (11): 4682-8. PubMed PMID: 14578386 .; Aragona et al., Long term treatment with sodium hyaluronate-containing artificial tears reduces ocular surface damage in patients with dry eye. Br J Ophthalmol . 2002 Feb; 86 (2): 181-4. PubMed PMID: 11815344., PubMed Central PMCID: PMC1771021], CMC compositions of the invention may be used to treat and prevent dry eye syndrome.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for treating and preventing dry eye syndrome, comprising a composition containing the carboxymethyl chitosan (CMC) as an active ingredient.

The pharmaceutical composition of the present invention is characterized in that it is a topical application formulation.

The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier in addition to the carboxymethyl chitosan (CMC), the active ingredient. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

Suitable dosages of the pharmaceutical compositions of the invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex of the patient, degree of disease symptom, food, time of administration, route of administration, rate of excretion and response to reaction. In general, the skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment. On the other hand, the dosage of the pharmaceutical composition of the present invention is preferably 0.001-2000 mg / kg body weight per day.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, may be administered by intravenous infusion, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like. It is preferable that the route of administration of the pharmaceutical composition of the present invention is determined according to the type of the disease to be applied. Since carboxymethyl chitosan (CMC), an active ingredient in the pharmaceutical composition of the present invention, is easily degraded in the body, a topical application mode of administration in which the composition can be directly administered to the epithelium is preferred.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

In the pharmaceutical composition of the present invention, the topical application formulation is an eye drop, cream, ointment, gel, or lotion.

According to one aspect of the invention, the invention provides a method for screening dry eye prophylaxis and treatment material comprising the following steps:

a) topically applying any agent to the test animal; And

b) determining whether caspase-3 and PARP positive cell numbers in corneal epithelial and conjunctival epithelium are reduced or TNF-α positive cell numbers in corneal stroma and conjunctival mucosa compared to control without any of the above agents. When the number of cells is reduced, it is identified as a material for preventing and treating dry eye syndrome.

More preferably, in addition to the reduction of the cells may further include determining whether to reduce the amount of tear reduction, fluorescein, rose bangal staining site that appears upon corneal injury.

As described above, according to the present invention, the composition for treating and preventing dry eye syndrome containing CMC has an excellent therapeutic effect against dry eye compared to conventional commercial hyalic acid solutions. More specifically, when the CMC composition of the present invention was treated in a test animal that caused dry eye syndrome, it inhibited the inflammatory and apoptotic reactions occurring after dry eye induction, and promoted recovery of damaged epithelial cells of the cornea and conjunctiva. Rather, it promoted the regeneration of goblet cells that secrete the mucus of the conjunctiva. In addition, the tear volume reduced by dry eye syndrome was restored to the normal tear range. Therefore, compared with the commercially available hyalic acid solution, the effect is not inferior at all, and it is considered that carboxymethyl chitosan, which shows a somewhat superior biological effect, is very likely as a therapeutic agent for dry eye drops. Therefore, if the dry eye treatment using the carboxymethyl chitosan composition according to the present invention is successfully commercialized, domestic economic activation and related pharmaceutical industry growth is expected by the demand for dry eye treatment worldwide, and various eye diseases caused by dry eye are expected. It is expected to prevent economic loss caused by disease and decreased vision and ultimately contribute to maintaining a comfortable life of human.

Figure 1 shows the results of measuring the STT change in dry eye animal model groups: control; Sterile distilled water treatment, HA group; 0.1% sodium hyaluronate treatment, CMC group; 0.5% CMC treatment (* (p <0.05), ** (p <0.001) means significant difference from control).
Figure 2 is the result of measuring the change in the fluorescein score (fluorescein score) of dry eye animal model groups: control; Sterile distilled water treatment, HA group; 0.1% sodium hyaluronate treatment, CMC group; 0.5% CMC treatment (** (p <0.001) means significant difference from control).
Figure 3 is the result of measuring the change in Rose bengal score of dry eye animal model groups: control; Sterile distilled water treatment, HA group; 0.1% sodium hyaluronate treatment, CMC group; 0.5% CMC treatment (** (p <0.001) means significant difference from control).
4 shows histopathological test results (corneas) of dry eye animal model groups: control; Sterile distilled water treatment (A-C), HA group; 0.1% sodium hyaluronate treatment (D-F), CMC group; 0.5% CMC treatment (G to I) (Scale bars = 80 μm, Dot arrow represents measured epithelial thickness, arrow represents measured total corneal thickness).
Figure 5 shows histopathology test results (conjunctival) of dry eye animal model groups: control; Sterile distilled water treatment (A-C), HA group; 0.1% sodium hyaluronate treatment (D-F), CMC group; 0.5% CMC treatment (G to I) (Scale bars = 80 μm, arrow indicates measured epithelial thickness).
Figure 6 shows the immunohistopathology test results of dry eye animal model groups, showing the change in the number of caspase-3 positive cells detected in corneal and conjunctival epithelial cells: control; Sterile distilled water treatment (A and B), HA group; 0.1% sodium hyaluronate treatment (C and D), CMC group; 0.5% CMC treatment (E and F) (Scale bars = 80 μm).
Figure 7 shows the immunohistopathology test results of dry eye animal model groups, showing the change in the number of PARP positive cells detected in corneal and conjunctival epithelial cells: control; Sterile distilled water treatment (A and B), HA group; 0.1% sodium hyaluronate treatment (C and D), CMC group; 0.5% CMC treatment (E and F) (Scale bars = 80 μm).
Figure 8 shows the immunohistopathology test results of dry eye animal model groups, showing the change in the number of TNF-α positive cells detected in cornea stroma and conjunctiva lamina propria: control ); Sterile distilled water treatment (A and B), HA group; 0.1% sodium hyaluronate treatment (C and D), CMC group; 0.5% CMC treatment (E and F) (Scale bars = 80 μm).

Hereinafter, the present invention will be described in more detail with reference to the following examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example  1. Production of dry eye animal model

Fifteen healthy Beagle dogs (adult males) weighing 8.7 ± 1.9 kg and 18.2 ± 2.8 months of age were used. Each test animal was subjected to ambient temperature (20-25 ° C.), 40-45% relative humidity, and a 12 hour light cycle. Breeding in cages standardized under light-dark cycle conditions, they provided a standard diet and tap water. Care and handling of all the experimental animals was performed in accordance with the Association for Research in Vision and Ophthalmology (ARVO) method for the use of animals in ophthalmology and vision research, and also approved by Kyungpook National University Experimental Animal Ethics Committee. The 15 Beagle dogs were divided into three groups (control group, HA group, and CMC group) to be composed of 5 animals each, and one eye of each animal was randomly selected. Dry eye animal models were induced in all groups according to the following steps.

Step 1: Four weeks, four times daily, 0.4% BAC (Sigma Aldrich Co., St. Louis, Mo, USA) eye drops were administered to the eyes of animals. On day 14 of starting 0.4% BAC eye drops dry eye conditions were induced in all animals. Then, step 2: treatment was started for 2 weeks. In the control group, only sterile distilled water was applied, and in the HA group, 0.1% sodium hyaluronate (Lacure., Samil pharm. Co., Ltd. Kyunggi-do.Korea) eye drops were applied, and the CMC group was 0.5%. CMC (carboxymethyl chitosan) (Chitopol., Chembio. Co., Ltd. Chungchengbuk-do.Korea) eye drops were applied. Treatment of eye drops in each group was applied 2 hours after instillation of 0.4% BAC, respectively.

Example  2. Ophthalmic Test Analysis and Results

The eyes of the dry eye animals were examined daily with naked eyes and as ophthalmological tests on days 0, 3, 7, 10 and 14 of step 2. Diet and water were not fed 12 hours before the test. For the calibration of animals, medetomidine (Domitor., Orion corporation animal health., Finland, 20 mg / kg, IV) and ketamine (Ketamine., Yuhan Corp., Korea, 25 mg / kg, IV) were applied to all animals. It was.

2-1. Aqueous tear production measurement

The Chirmer tear test (STT) was performed by measuring tears secreted in response to conjunctival stimulation and basal non-reflective secretion. This was performed using a schimer strip, and secreted tears were measured with a filter paper 0.5 mm wide and 3.5 mm long. The strip was inserted into the ocular surface at the lateral canthus for 1 minute. The wet area was measured in millimeters according to the extent stamped on the strip. Normally, the normal range of dogs is 15-25 mm / min and less than 10 mm / min when dry.

As shown in FIG. 1, the Schirmer score of eyes treated with carboxymethyl chitosan (CMC) was significantly higher than the control (treated with sterile distilled water) on days 7, 10 and 14 of treatment. (p <0.05). On the other hand, there was no significant difference in two-way ANOVA between the CMC group and the artificial tear treated group. On day 7, 10 and 14 days of treatment, the mean STT value of each group was 10.40 ± 1.140, 8.40 ± 0.548 and 7.80 ± 0.447 of control group, 12.60 ± 4.037, 15.20 ± 1.789 and 16.60 ± 2.702 of HA treatment group, respectively. 15.80 ± 1.304, 16.20 ± 0.837 and 20.00 ± 1.871.

2-2. cornea Fluororesin  dyeing( Corneal fluorescein staining )

Corneal fluorescein staining was performed with a slit lamp microscope (SM-70N) under cobalt blue light 1 minute after topical application of 1% sodium fluorescein strip into the conjunctival sac. , Takagi Seiko.Co., Ltd. Nagano.Japan) and evaluated using an attached digital camera (EOS450D., Cannon. Co., Ltd. Tokyo. Japan).

Corneal fluorescein staining is classified into a grading system based on the stained area of the cornea and is divided into 0-4 grades. The entire area of punctate staining is designated as Grade 0 if it is not dot staining and Grade 1 if stained to 1/8 or less. Grade 2 is dyed 1/4 or less, grade 3 is dyed 1/2 or less, grade 4 is dyed at least 1/2 or the entire area.

As shown in FIG. 2, on days 3, 7, 10 and 14 of treatment, the fluorescein score of the group treated with CMC was significantly reduced compared to the control (treated with sterile distilled water only) (p < 0.05). However, there was no significant difference between the CMC group and the HA group. The mean corneal fluorescein staining values of each group at 3, 7, 10 and 14 days of treatment were 3.60 ± 0.548, 4.00 ± 0.000, 3.80 ± 0.447 and 3.80 ± 0.447 in the control group, and 2.40 ± 0.548, 1.40 ± in the HA treatment group, respectively. 0.548, 1.10 ± 0.547 and 0.80 ± 0.273, CMC treatment groups were measured as 2.00 ± 0.000, 1.00 ± 0.000, 0.80 ± 0.273 and 0.70 ± 0.273.

2-3. cornea rose Bengal  dyeing( Corneal rose bengal staining )

Rose Bengal staining was evaluated using a slit lamp microscope under green filter light 1 minute after topical application of 2 μl 1% Rose Bengal solution into the conjunctival sac and photographed with a digital camera.

The ocular surface was examined and graded with a Van Bijsterveld grading system. The grading system is divided into three regions (nasal, temporal ocular conjunctiva and cornea) in each eye. The intensity of rose bengal staining is divided into a range of 0 to 3 for each region. The maximum value of staining for each eye is 9 for humans. However, in the present invention, only the extent of the cornea was evaluated because of the difference in external morphology between the human eye and the dog's eye. Therefore, the maximum value of staining for each eye in the present invention was set to 3.

As shown in FIG. 3, on day 7, 10 and 14 days of treatment, lodge bengal staining values of the group treated with CMC were significantly reduced (p <0.05) compared to the control (treated with sterile distilled water only). In addition, compared with the HA group, the CMC treatment group showed a further decrease effect from day 7. At 7, 10 and 14 days of treatment, the mean corneal rose bengal staining values of the control groups were 3.00 ± 0.000, 3.00 ± 0.000 and 2.60 ± 0.548, and 2.20 ± 0.447, 1.60 ± 0.548 and 1.40 ± 0.548, CMC for the HA treatment group, respectively. Treatment groups were measured as 1.60 ± 0.894, 1.20 ± 0.447 and 1.00 ± 0.000.

2-4. Histopathology test

At the completion of step 2, test animals were euthanized by overdose (250 mg / kg or more) of pentobarbital sodium. Eyeballs are then extracted and fixed with Davidson's solution (37.5% ethanol, 12.5 acetic acid and 25% formaldehyde), and the palpal conjunctiva is taken from the same location and 10% neutral buffered formalin (neutral buffered). formalin) for at least 24 hours. Subsequently, they were embedded with paraffin and mounted on a paraffin block to cut each of the three corneal and conjunctiva into 2-3 μm sections, each of which was a single slide. ). For histopathology and periodic acid Schiff (PAS) staining of goblet cells producing mucous, the respective slides were stained with hematoxylin and eosin. Histological evaluation was performed in the central area of the cornea or conjunctiva.

As shown in FIG. 4, cornea epithelial damage and the number of inflammatory cells infiltrated into the stroma were significantly reduced in the HA treated group and the CMC treated group compared to the control group (p <0.01), and also corneal epithelium The thickness of was significantly increased in both HA and CMC groups compared to the control. In addition, as shown in FIG. 5, the conjunctiva increased epithelial thickness in both HA and CMC groups compared to the control group, and the number of inflammatory cells infiltrated into the mucosa (lamina propria) decreased and goblet cells. The number of was increased in both groups (p <0.01).

2-5. Immunohistopathologic test

Citrate buffer antigen (epitoe) retrieval pretreatment was performed first, followed by deparaffinization of the histological paraffin blocks of the prepared cornea and conjunctiva. Briefly, a staining dish containing 10 mM citrate buffer (pH 6.0) and a constant temperature bath are pre-heated until they reach 95 to 100 ° C., soaking the tissue preparation slide in the staining dish, and The lid was loosely placed on top. The thermostat of the bath was turned off and incubated for 20 minutes. The dye dish was then placed at room temperature and the slides were allowed to cool for 20 minutes. After restoring the epitope, the compartments were immunostained.

As shown in FIGS. 6 and 7, the caspase-3 and PARP positive cell numbers were significantly reduced in corneal and conjunctival epithelial cells of HA and CMC treated groups compared to controls (p <0.01), as also shown in FIG. 8. Likewise, the number of TNF-α positive cells was significantly reduced in both cornea stroma and conjunctiva lamina propria of HA, CMC group compared to control group (p <0.01).

Claims (8)

A composition for treating and preventing dry eye syndrome containing carboxymethyl chitosan (CMC) as an active ingredient.
The method of claim 1, wherein the carboxymethyl chitosan (CMC) is a composition for treating and preventing dry eye, characterized in that the polysaccharide (polysaccharide) derived from chitin (chitin).
The composition of claim 1, wherein the composition increases epithelial thickness of the cornea and conjunctiva and reduces the number of inflammatory cells.
The method of claim 1, wherein the composition reduces the number of caspase-3 and PARP positive cells in corneal epithelial and conjunctival epithelial cells, and reduces the number of TNF-α positive cells in corneal stroma and conjunctival mucosa. And prophylactic compositions.
A pharmaceutical composition for treating and preventing dry eye, comprising the composition of claim 1.
6. The pharmaceutical composition of claim 5, wherein the composition is a topical application formulation.
The pharmaceutical composition of claim 6, wherein the topical application formulation is an eye drop, cream, ointment, gel or lotion.
A method for screening dry eye prophylaxis and treatment comprising the following steps:
a) topically applying any agent to the test animal; And
b) determining whether caspase-3 and PARP positive cell numbers in corneal epithelial and conjunctival epithelium are reduced or TNF-α positive cell numbers in corneal stroma and conjunctival mucosa compared to control without any of the above agents. When the number of cells is reduced, it is identified as a material for preventing and treating dry eye syndrome.

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