JP2000319194A - Agent for therapy of dry keratopathy - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an agent for the therapy of dry keratopathy which enables the onset of cornea inflammatory disease (especially dry keratopathy) to be prevented and treated without causing a side effect such as stimulation in a topically administered part by including human serum albumin having a specific concentration. SOLUTION: This therapeutic agent comprises human serum albumin in a concentration of 0.001-0.3 mg/mL which preferably shows pH 6.0-7.5 and an osmotic pressure ratio of 0.5-1.5 based on physiological salt solution. The composition as an agent for the therapy of dry keratopathy comprises e.g. 0.001-0.3 (wt/vol)% human serum albumin, 0. 15-2.0 (wt/vol)% glucose, 80-140 m equivalent sodium ion, 4-25 m equivalent potassium ion, 0-4 m equivalent magnesium ion, 0-0.05 m equivalent borate ion, 10.0-150 m equivalent chloride ion, 0-0.03 (wt/vol)% sodium acetyltryptophane and 0-0.15 (wt/vol)% sodium caprylate, and the quantity of instillation is preferably 20-60 μL/(one eye)/(one dose) and is administered at a frequency of 3-10 times a day.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an agent for preventing and treating the occurrence of inflammatory diseases of the cornea, particularly dry keratosis, and more particularly, to a drug which is not administered to the eye to cause irritation or corneal disorder. The present invention relates to a therapeutic agent for dry corneas containing serum albumin as a main component, which has characteristics.

[0002]

2. Description of the Related Art Dry keratosis is a disease caused by drying of the cornea, and is one of the diseases of the eye that has been rapidly increasing in recent years. Hormonal involvement is suspected because it occurs frequently in middle-aged humans, especially in menopausal women, but the cause has not yet been fully elucidated. The cause of the increase in the number of patients is a change in the environment, that is, a society that overuses the eyes. Prolonged use of a computer, driving a car, watching TV for a long time, etc., reduces the number of blinks and thus causes xerokeratosis. In addition, it has been found that dry keratopathy is likely to be caused by an air environment such as drying of a room by cooling and heating. Furthermore, it is said that dry keratosis occurs as a symptom of Sjograin's syndrome or as a complication of keratoconjunctivitis. The subjective symptoms of patients with these diseases include eye fatigue, foreign body sensation, and burning sensation, which are obstacles to the patient's normal life.

[0003] There is no causal treatment as a treatment for dry keratosis, and symptomatic treatment is currently performed. Among these, non-drug treatments include punctal closure, ocular suture, and non-hydrous soft contact lenses. However, these treatments are often uncertain, or the irritation of the eyes becomes so severe that the treatment must be discontinued. As a drug treatment method, there is a method using hyaluronic acid, chondroitin sulfate, artificial tear, or the like. However, the effect of hyaluronic acid is weak, and the effect of artificial tears is extremely temporary and is not sufficient. In addition, despite the fact that drugs such as hyaluronic acid are therapeutic agents for keratosis xerosa, keratosis such as pruritus, irritation, hyperemia, and superficial keratitis appears as side effects in addition to blepharitis and eyelids Sometimes. For this reason, there are few cases where symptoms are completely asymptomatic even when treated with a drug such as hyaluronic acid, and as a result, patients may be disappointed.

[0004] Recently, a drug containing albumin as an active ingredient and useful for treating symptoms such as keratoconjunctival disorder and dry eye has been reported (WO97 / 39769). However,
The pharmaceutical composition contains high amounts of albumin, especially 10-100
It is a pharmaceutical composition also containing 0 mg / ml. Thus, a drug containing a large amount of albumin, when instilled as an aqueous solution, has a high osmotic pressure,
May cause corneal damage. Also, the viscosity becomes high, and it becomes difficult to handle as an eye drop.

[0005] On the other hand, albumin is generally known as a stabilizer for protein preparations and enzyme preparations, and as eye drops using albumin as a stabilizer, those containing interferon, fibronectin or lysozyme as an active ingredient are known. (Japanese Unexamined Patent Publication No. 6-271478,
5-310592, JP-A-61-103836, JP-A-57-12
No. 6422). However, it is not clear from a conventional eye drop using albumin as a protein stabilizer that albumin itself exerts a therapeutic effect on dry keratitis.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a drug for preventing and treating the occurrence of inflammatory diseases of the cornea, particularly dry keratosis, without causing side effects such as irritating action at the administered topic. It is in.

[0007]

Means for Solving the Problems In order to achieve the above object, the present inventors have conducted intensive studies and found that a drug prepared at a low concentration of high-purity serum albumin has a stimulating effect on the administered local topic of the eye. The present inventors have found excellent effects on keratopathy without producing the side effects of the present invention, and have reached the present invention.

[0008] That is, the present invention is a therapeutic agent for dry keratosis comprising human serum albumin having a human serum albumin concentration of 0.001 to 0.3 mg / ml as an active ingredient.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The human serum albumin used in the present invention includes albumin purified from human plasma and purified or recombinant human serum albumin produced by genetic engineering. It is desirable that the human serum albumin be of high purity. Specifically, the purity is desirably 99% or more. Impurities become foreign antigens and can cause allergic reactions or foreign inflammation, which can delay or exacerbate the treatment of xerokeratosis. Also,
Recombinant human serum albumin is practically preferable because there is no possibility of contamination by a heat-resistant virus or a Creutzfeldt-Jakob pathogen.

As the solubilizing agent for human serum albumin, a solvent applicable to the eye is preferable, and examples thereof include physiological saline, isotonic phosphate buffer, artificial tears and the like. The concentration of human serum albumin needs to be 0.001-0.3 mg / ml. The concentration of human serum albumin was 0.3 mg /
Even if the amount exceeds 100 ml, the effect of suppressing the onset of dry keratitis is not enhanced, but an irritating effect on the eyes is produced.
Further, when the amount is less than 0.001 mg / ml, the inhibitory effect on keratosis xerostomia is not exhibited.

The therapeutic agent for xerokeratosis of the present invention must be isotonic with normal tears, blood and the like, and have a neutral pH. If there is a great difference between them, eye pain or infiltration of the eye during instillation causes discomfort. In addition, physical or chemical irritation may worsen eye diseases or cause new eye disorders. Since the concentration that is isotonic with the physiological osmotic pressure of human serum albumin is about 5 w / v%, as a therapeutic agent for keratosis sicca, at an albumin concentration of about 5 w / v% or more, a physiologically hypertonic solution is used. It causes damage to the cornea and delays the treatment of xerokeratosis. Therefore, in the present invention, the pH of the therapeutic agent for dry cornea is adjusted to pH 6.0.
To 7.5, preferably 6.8 to 7.4. In addition, the osmotic pressure ratio (physiological saline solution ratio) is in the range of 0.5 to 1.5, preferably 0.8 to 1.0.

The solute in the therapeutic agent for dry keratosis of the present invention is a solution applicable to ophthalmology, and comprises sodium chloride,
It is an aqueous solution containing some or more of glucose, phosphoric acid, boric acid, sodium bicarbonate, sodium carbonate, potassium chloride, calcium chloride and magnesium chloride. Further, the therapeutic agent for xerokeratosis of the present invention does not contain proteins other than human serum albumin, or contains trace amounts of other proteins. It can also contain sugars such as hyaluronic acid, carboxymethylcellulose and glucose.

Further, the therapeutic agent for xerokeratosis of the present invention promotes the onset of xerokeratosis, delays the treatment thereof,
Alternatively, they may contain unobstructed concentrations of albumin stabilizers, antibacterial agents, preservatives and the like. As an albumin stabilizer, sodium acetyltryptophan,
Examples include sodium caprylate and the like. The amount of sodium acetyltryptophan added is, for example, 1 to 4 w / w% with respect to recombinant human serum albumin, preferably
2 to 2.25% w / w. The amount of sodium caprylate added is, for example, based on recombinant human serum albumin.
It is 0.5-3 w / w%, preferably 1-2 w / w%. Antibacterial substances include antibiotics, sulfa drugs and quinolone compounds. Antibiotics include sulbenicillin sodium, pimaricin dibekacin sulfate,
Gentamicin sulfate, Becamycin sulfate, Lactopion
III erythromycin, and chloramphenicol. Examples of the sulfa drug include sulfisoxazole and the like, and quinolones include ofloxacin. Examples of the preservative include parahydroxybenzoic acid and salts thereof, esters of parahydroxybenzoic acid, lower alkyl ester preservatives, and benzalkonium hydrochloride.

The container for containing the therapeutic agent for dry keratosis of the present invention includes a glass or plastic container having one or two or more chambers for storing the drug. For example, when a container having two or more chambers is used, a component containing human serum albumin is filled in one chamber, and another component is filled in another chamber. Further, the dosage form of the therapeutic agent for dry keratosis of the present invention is human serum albumin alone or in a solid or solution state in combination with other components, and other components not containing human serum albumin are similarly solid or solution. It is good also as a state. The solid state includes powder, fine granules, tablets and the like, and the solution state is a solution dissolved in distilled water or an aqueous solvent. In the case of instillation, if it consists of two or more liquid preparations, it is administered after mixing, and if it contains a solid component, it is administered after dissolving and mixing the solid component in a solvent.

As for human serum albumin, a sterile bulk powder is used, and the above-mentioned container is aseptically filled. After dissolving the non-sterile bulk powder in a water solvent, it is necessary to perform a heat treatment, for example, at 60 ° C. for 10 hours, or a sterilization treatment such as filtration sterilization using a membrane filter having a pore size of 0.22 μm.

One example of the therapeutic agent for xerokeratosis of the present invention has the following composition.

[0017]

[Table 2]

The therapeutic agent for xerokeratosis of the present invention is administered topically to the eye, that is, by instillation. Clinically, the amount of eye drop is 20 to 60 μl / eye at a time, preferably 30 to 50 at a time.
μl / eye, and preferably 3 to 10 times, preferably 4 to 6 times per day.

[0019]

EXAMPLES Hereinafter, the therapeutic agent for dry corneas of the present invention will be described in detail with reference to examples. Preparation Example 1 A therapeutic agent for dry corneas having the following composition was prepared. Human serum albumin 0.1 mg artificial tears: glucose 28 mg sodium chloride 55 mg potassium chloride ion 16 mg sodium monohydrogen phosphate 33 mg acetyltryptophan sodium 0.24 mg sodium caprylate 0.135 mg distilled water for injection 10.0 ml

The above compound is dissolved in distilled water for injection, and p
After adjusting to H7.0 to pH7.5, the mixture was filled in a container purged with nitrogen, and then heat-sterilized at 60 ° C. for 10 hours. The resulting solution was filled into a container that was opaque to light and oxygen and carbon dioxide gas. Albumin concentration was 0.001 w / v%. The osmotic pressure ratio (physiological saline solution ratio) is 0.95-1.
05.

Preparation Example 2 Two kinds of solutions A and B having the following compositions were prepared. A solution: sterile human serum albumin 0.1 mg acetyltryptophan sodium 0.24 mg sodium caprylate 0.135 mg distilled water for injection 1.0 ml B solution: glucose 38.4 mg sodium chloride 55 mg potassium chloride ion 15 mg dry sodium carbonate 5.6 mg phosphorus Sodium hydrogen oxylate 6mg Distilled water for injection 9.0ml

In one of the two plastic containers,
The solution B was filled and sterilized at 120 ° C. for 20 minutes. Then, after filling the other chamber with nitrogen gas, the solution A was charged and heat-treated at 60 ° C. for 10 hours. At the time of use, the A solution and the B solution were mixed. Albumin concentration is 0.001 w / v
%, And the osmotic pressure ratio (physiological saline solution ratio) is 0.95 to 0.95%.
1.05.

Preparation Example 3 A mixture (all the following components are sterile): human serum albumin 0.1 mg acetyltryptophan sodium 0.24 mg sodium caprylate 0.135 mg glucose 22 mg sodium chloride 85 mg potassium chloride ion 13 mg dry sodium carbonate 6 mg phosphoric acid Sodium monohydrogen 27.5mg B solution: distilled water for injection 10.0ml

One of the two plastic containers was filled with nitrogen gas. The solution B was filled into one room filled with nitrogen, and sterilized at 120 ° C. for 20 minutes. At the time of use, A tablet was dissolved in B solution. Albumin concentration is 0.001w /
v%, osmotic pressure ratio (physiological saline solution ratio) is 0.95 to 1.0
It was 5. pH was 6.8.

Test Example 1 The effect of human serum albumin dissolved in physiological saline on dry keratosis was confirmed using Japanese white male rabbits (body weight 2-3 kg). First, a rabbit is fixed in a fixed box under urethane anesthesia, the eyelid is forcibly opened using a retractor, 50 μl of a test solution is dropped on the ocular surface (corneal surface) of one eye, and left as it is for 6 hours. did. Thereafter, 50 μl of a 1% methylene blue solution dissolved in a physiological saline solution was dropped, and two minutes later, the ocular surface was washed twice with a physiological saline solution. Then, the rabbit was exsanguinated and killed under Nembutal anesthesia, the cornea was excised, immersed in an acetone solution (100 W / V% acetone solution: saturated sodium sulfate aqueous solution = 7: 3, volume ratio) for 24 hours, and the eluted dye concentration was measured. The absorbance was measured at 660 nm. Test drug solution, as described in Preparation Examples 1-3,
It was prepared by dissolving human serum albumin in physiological saline to a predetermined concentration. Artificial tears, which are positive control drugs, use commercially available Mytayer (manufactured by Takeda Pharmaceutical Co., Ltd.), and chondroitin sulfate is chondron (containing 3W / V% chondroitin sulfate,
For hyaluronic acid, hyaluronic acid (containing 0.1 W / V% hyaluronic acid, manufactured by Santen Pharmaceutical Co., Ltd.) was used as it was or diluted to a predetermined concentration with a physiological saline solution. Table 3 shows the results.

[0026]

[Table 3] Effect of human serum albumin dissolved in physiological saline *; Significantly different at p <0.05 compared to the solvent group. **; significantly different at p <0.01 compared to the solvent group.

As is clear from Table 3, the concentration of human serum albumin at a concentration of 0.001 to 0.3 (W / V)% significantly suppressed the occurrence of experimental xerokeratosis. It showed a stronger effect than the agents chondroitin sulfate and hyaluronic acid.

Test Example 2 The effect of human serum albumin dissolved in an isotonic phosphate buffer was confirmed using Japanese white male rabbits (body weight 2-3 kg). The test was performed in the same manner as in Test Example 1. The test compound solution was prepared so that human serum albumin had a predetermined concentration.
It was prepared by dissolving in an isotonic phosphate buffer.

[0029]

TABLE 4 Effect of human serum albumin dissolved in isotonic phosphate buffer **; significantly different at p <0.01 compared to the solvent group.

As is clear from Table 4, even if human serum albumin was dissolved in an isotonic phosphate buffer, 0.001 (W / W
At a concentration of V)% or more, the development was significantly suppressed for experimental keratopathy.

Test Example 3 The test was performed in the same manner as in Test Example 1 except that 3 to 6 Japanese white male rabbits (body weight: 2 to 3 kg) were used per group. The test compound solution was prepared by dissolving human serum albumin in the artificial tear formulation of Preparation Example 1 to a predetermined concentration.

[0032]

[Table 5] Effect of human serum albumin dissolved in artificial tears *; Significantly different at p <0.05 compared to the solvent group. **; significantly different at p <0.01 compared to the solvent group.

As is evident from Table 5, even when dissolved in artificial tears, human serum albumin significantly inhibits the occurrence of experimental keratosis at a concentration of 0.001 (W / V)% or more. Was.

Test Example 4 The irritancy of albumin to tissues was examined as follows. First, 0.01,0.
1, 0.3, 1.0, 0.5, 0.10, 10.0, 2
A 5.0 w / v% human serum albumin solution and 0.2 ml of physiological saline were administered intradermally to the back skin of the rat, and immediately thereafter, 2 ml / kg of a 2% pontamine sky blue solution was intravenously administered. Forty-five minutes later, the animals were killed by decapitation, and a certain area of the intradermally stained portion of the back skin tissue was extracted, and the method of Katayama et al. (Katayama S., et al: Microbiol. Immunol., 22, 8)
9-101 (1978)). That is, 1 mL of 1.2N potassium hydroxide solution was added to the tissue piece, and 37 mL
After heating at 24 ° C. for 24 hours to dissolve the tissue, acetone / 0.1.
9 mL of a 6N phosphoric acid mixture (13/5) was added, and the mixture was shaken to extract and centrifuged. The dye concentration of the supernatant was measured with a spectrophotometer.

[0035]

[Table 6] Stimulating action of human serum albumin and hyaluronic acid on tissues

*: P compared to physiological saline group (solvent group)
<0.05, significant difference. **; p <0.01 compared with the physiological saline group (solvent group), with a significant difference.

As shown in Table 6, human serum albumin did not show a significant effect at 0.01 to 0.3 w / v%, but at a concentration of 0.3 w / v% or more, the leakage dye concentration was significant. As a result of the increase, it was found that the compound had a local stimulating effect. These facts suggest that human serum albumin may have an irritating effect on the eye at 0.3 w / v% or more.
It can be determined that it is appropriate that the ratio is not more than 01 w / v%.

[0038]

EFFECT OF THE INVENTION The therapeutic agent for xerokeratosis of the present invention has a feature that, by containing a low concentration of serum albumin, the occurrence of experimental xerokeratosis is significantly suppressed without exhibiting a local stimulating effect.

Claims (3)

[Claims] (1) a human serum albumin concentration of 0.001 to
A therapeutic agent for keratosis xerosclerosis, comprising human serum albumin at 0.3 mg / ml as an active ingredient. 2. The therapeutic agent for dry keratosis according to claim 1, wherein the pH is 6.0 to 7.5 and the osmotic pressure ratio (physiological saline ratio) is 0.5 to 1.5. 3. The therapeutic agent for dry keratosis according to claim 1, which has the following composition. [Table 1]