KR20030023098A - The curing composition for retina injury with sulfasalazine, and the producing method of thereof - Google Patents

Abstract

PURPOSE: Provided are sulfasalazine(SSZ) represented in the formula(1) or its water soluble salt of the formula(2) contained in an F eye drop composition as an active ingredient for treating retinal damage and a method of its preparation. The eye drop is used in treating retinal ischemia caused by glaucoma, diabetic retinal damage or degradation of retinal nerve due to uveitis. CONSTITUTION: The preparation method comprises: adding 0.8-0.9wt.% of NaOH at room temperature to an aqueous CH3CN solution with a ratio of 1:10(v/v, CH3CN/H2O); adding 8.5-9.5wt.% of sulfasalazine and reacting for 30 minutes at room temperature; freeze drying at -78deg.C for 48 hours to obtain a Na salt; and dissolving it in distilled water to make a 1-5mM concentration.

Description

Eye drops composition for the treatment of retinal injury, characterized in that it contains sulfasalazine as an active ingredient, and a method of manufacturing the same {THE CURING COMPOSITION FOR RETINA INJURY WITH SULFASALAZINE, AND THE PRODUCING METHOD OF THEREOF}

The present invention relates to an ophthalmic composition for treating retinal damage and a method for producing the same, comprising sulfasalazine (hereinafter referred to as SSZ abbreviation) or a water-soluble salt of sulfasalazine as an active ingredient.

Sulfasalazine (Sulfasalazine: C ₁₈ H ₁₄ N ₄ O ₅ S) represented by the following Chemical Formula 1 is a known substance and is used for the treatment of colitis and enteritis as a nonsteroidal inflammatory agent.

<Formula 1>

Applicant of the present invention filed on April 21, 2000, PCT / KR00 / 00378 (COMPOSITION AND METHOD FOR INTERVENING NEURONAL DEATH USING SULFASALAZINE) regarding the use of sulfasalazine as a therapeutic agent for neurological diseases.

Sulphasalazine is poorly soluble in solvents such as water, benzene, chloroform and ethers and is soluble in small amounts in alcohol.

Meanwhile, there is a need for the development of new drugs that protect the retina and treat retinal damage.

Korean Patent Application No. 10-1998-0702671 (PCT / JP1996 / 02850) (carbostyryl derivative for the treatment of ophthalmic diseases) is known ophthalmic disease therapeutic agent containing the carbostyryl derivative as an active ingredient, but with the active ingredient The composition is different, and the purpose and effect are different.

In addition, Korean Patent Application No. 10-2000-7008301 (PCT / JP1999 / 00369) (eye drops composition) comprises a specific compound or aldose reductase inhibitor as an active ingredient, characterized in that the treatment of diabetic corneal disorders Ophthalmic compositions for use in the treatment of the known are known, but the active ingredient is different from the present invention, the purpose and effect are different.

It is an object of the present invention to provide a novel active ingredient for treating retinal damage, a composition thereof, and a method of preparing the drug, which can be easily and repeatedly administered and can deliver the drug to a target site without undergoing hepatic metabolism.

1 is a process chart of the sulfa salinated sodium salt

Figure 2 is a photograph of the retinal protective effect according to the administration of the eye drops composition of the present invention in the rat vitreous cavity

GCL: Ganglion cell layer INL: Inner nuclear layer

Figure 3 is a quantitative analysis of the effect of administration of eye drops composition concentration

Figure 4 is a photograph of the retinal protection effect according to the administration of the eye drop composition of the present invention intraperitoneally

Figure 5 is a photograph of the retinal protection effect according to the administration of the eye drops composition of the present invention to the eyes of white rat

The present invention relates to an ophthalmic composition for treating retinal injury and a method for producing the same, comprising sulfasalazine or a water-soluble salt thereof as an active ingredient.

The present inventors have found that sulfasalazine exhibits a high effect when applied to the treatment of retinal disease and has been completed through several experiments.

Sulfasalazine is poorly soluble in water, so its solubilization is required for use as a medicament.

Soluble sulfasalazine is used in the form of water-soluble and harmless salts such as sodium salts and potassium salts.

A method for preparing sulfasalated sodium salt is described with reference to FIG. 1.

Acetonitrile (CH ₃ CN) is dissolved in distilled water at a ratio of 1:10 (v / v) at room temperature to form an aqueous solution.

Sodium hydroxide (NaOH) was mixed in an aqueous acetonitrile solution with 0.8 ~ 0.9 wt%, sulfasalazine was added in an amount of 8.5 to 9.5 wt%, reacted at room temperature for 30 minutes, and the reaction solution was dried at a low temperature of -78 ° C for 48 hours, A pale orange sulfasalated sodium salt is obtained.

The obtained sulfasalazine sodium salt is dissolved in distilled water at a concentration of 1 to 5 mM to prepare an ophthalmic composition for treating retinal injury, which comprises the sulfasalazine sodium salt of the present invention as an active ingredient.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples, but these examples do not limit the contents of the present invention.

Example 1 Preparation of Sulphasalazine Sodium Salt

30 ml of acetonitrile was dissolved in 300 ml of distilled water at room temperature (15-18 ° C.) to form an aqueous solution.

2.9 g of sodium hydroxide was mixed into an acetonitrile aqueous solution, and 30 g of sulfasalazine was added and reacted at room temperature for 30 minutes.

The reaction solution was dried at a low temperature of −78 ° C. for 48 hours to obtain 30 g of a pale orange sulfasalated sodium salt (C ₁₈ H ₁₃ N ₄ O ₅ SNa: molecular weight 420) powder.

<Example 2> Preparation of an ophthalmic composition containing sulfasalazine as an active ingredient

0.21 to 2.10 g of sulfasalin sodium salt prepared by the method of Example 1 were dissolved in 1 ml of distilled water at room temperature (15 to 18 ° C), respectively, and the concentration of sulfasalin sodium salt was 0.5 mM, 1 mM, 3 mM, and 5 mM. An ophthalmic composition for treating retinal injury was prepared.

Experimental Example 1 Retinal Ischemia Induction and Ocular Composition Administration in Rats

Male rats (Sprague Dawley, 8 weeks old, 300 g), 20 animals were used as experimental animals.

Rompun and ketala were mixed at a ratio of 1: 2 and anesthetized by intramuscular injection at 1 ml / kg.

Thirty minutes before ischemia, the ophthalmic composition prepared in Example 2 was administered at a concentration (0.5 mM to 5 mM) in 5 rats of 10 μl in each vitreous cavity.

The pressure inside the eyeball was increased to 160 mmHg using a pressure gauge with a pressure gauge to block the blood flowing to the retina for 1 hour to induce retinal ischemia.

After 24 hours of ischemia, the eye was removed, the retina was separated, fixed in 2.5% glutaraldehyde, and washed with phosphate buffer.

After dehydration in the order of increasing alcohol concentration, it was substituted with propylene oxide and embedded in an epon mixture.

Embedded tissue was made into 1 μm tissue sections and stained with toluidine blue, and then the retinal degeneration and protective effect were observed under an optical microscope.

This experimental result is shown in FIG.

It was observed in the cells (arrows and asterisks) that died in the retinas damaged by ischemia, but in the cells to which sulfasalated sodium salt was administered, the dead cells were significantly reduced.

<Quantitative effect of eye drop composition concentration>

Each tissue specimen of Experimental Example 1 was made, and the number of viable cells was counted in the range of 100 X 100 μm in each layer in the retina under an optical microscope, and expressed in% compared to normal.

The measured values were analyzed by student T-test (p <0.05) for statistical analysis, and the results are shown in FIG. 3.

When the ophthalmic composition of Example 2 was administered into the vitreous cavity, the retinal protective effect was observed at the concentration of sulfasalated sodium salt at 1 to 5 mM, and the highest protective effect was at 3 mM.

Retinal nerve cells remained 24% in the ganglion cell layer and 50% in the inner nucleus layer by ischemia, but 59.9% and 77.7% remained in the ganglion cell layer after sulfasalin 3 mM administration.

The results are summarized in Table 1 together with the results of Experimental Example 2 described below.

Experimental Example 2 Induction of Retinal Ischemia and Intraperitoneal Ocular Composition in Rats

Five rats were prepared.

Thirty minutes before the induction of retinal ischemia, 5 mg / kg of an ophthalmic composition having a concentration of 5 mM sulfasalin sodium salt prepared in Example 2 was injected into each abdominal cavity of the rat.

Retinal ischemia was induced in rats in the same manner as in Experimental Example 1.

The same amount of ophthalmic composition was administered to the abdominal cavity once more 30 minutes after ischemia.

In the experiment by intraperitoneal injection of sulfasalin sodium salt, the same result as in the intravitreal administration was obtained, and the results obtained after the intraperitoneal administration of the ocular composition 5 mg / kg are shown in FIG. 4 and Table 1.

The loss and degeneration of cells in the ganglion cell layer (GCL) and the inner nuclear layer (INL) due to ischemia are protected by sulfasal sodium salts.

Table 1 shows the results of Experimental Example 1 and Experimental Example 2 together.

<Table 1> Protective effect of retinal degeneration by the administration of eye drops composition

division Ganglion cell layer Inner layer n Intraluminal SSZ Administration Group 59.9 ± 8.7% 77.7 ± 5.3% 5 Intraperitoneal SSZ Administration Group 55.2 ± 6.2% 70.3 ± 7.1% 5 Control 24 ± 7.6% 50 ± 4.2% 5

Experimental Example 3 Experimental Retinal Protective Effects of Eye Drop Compositions Administered to the Eyes of Rats

Ten rats were used for the experiment.

The ophthalmic composition having a concentration of 3 mM sulfasalin sodium salt prepared in Example 2 was directly administered to the eyes of rats.

Two drops of 50 μl per drop were administered 6 times over 2 hours every 20 minutes.

Ischemia was induced in the retina of the rat in the same manner as in Experimental Example 1.

Again, two drops of the ophthalmic composition were administered every 20 minutes for 1 hour.

Retina was detached 24 hours after the final administration, and the effect of inhibiting retinal degeneration was investigated.

The results are shown in Fig. 5 and Table 2.

In the ischemia-damaged retina, cell loss was observed in the ganglion cell layer, and in the inner core layer, dying cells appearing as black spots.

However, the sulfasalized sodium salt-containing eye drops of the present invention contained about twice as many cells as the control group in the ganglion cell layer of the retina, and condensed and degenerated cells were reduced in the inner nuclear layer.

Table 2: Effect of Ophthalmic Composition Administration of the Present Invention

division Ganglion cell layer Inner layer n SSZ administration group 45.2 ± 5% 60.5 ± 5.3% 5 Control 27 ± 3.6% 53 ± 5.2% 5

In the ganglion cell layer, 27.0% of the cells remained in the control group, 45.2% of the cells in the ophthalmic composition-administered group of the present invention, 53.0% of the control group in the inner core layer, and 60.5% of the cells in the ocular composition-administered group of the present invention.

According to the present invention, there is provided an ophthalmic composition for treating retinal injury, and a method for producing the same, comprising sulfasalazine or a water-soluble salt thereof as an active ingredient.

The ophthalmic composition of the present invention is used for the treatment of retinal ischemia due to glaucoma, retinal damage due to diabetic retinopathy, retinal nerve degeneration due to uveitis, and the like.

Claims (5)

An ophthalmic composition for treating retinal injury, comprising sulfasalazine represented by the following formula (1) as an active ingredient, or a water-soluble salt thereof An ophthalmic composition for treating retinal injury, characterized by containing sodium salt of sulfasalazine represented by the following Chemical Formula 2 as an active ingredient: In acetonitrile aqueous solution prepared by dissolving acetonitrile in distilled water at a ratio of 1: 10 (v / v), 0.8% to 0.9% by weight of sodium hydroxide (NaOH) was added, and 8.5% to 9.5% by weight of sulfasalazine was added. The reaction solution was dried at a low temperature of -78 ° C. for 48 hours, and the sulfasalin sodium salt was dissolved in distilled water at a concentration of 1 to 5 mM, thereby preventing retinal damage. Method for preparing the ophthalmic composition for treatment The ophthalmic composition for treating retinal injury according to claim 1 or 2, wherein the retinal injury is ischemia of the retina due to glaucoma, retinal injury due to diabetic retinopathy, and degeneration of the retinal nerve due to uveitis. An ophthalmic composition for treating retinal injury, comprising the sulfasalin sodium salt prepared by the method of claim 3 as an active ingredient.