JP2006348025A - Prophylactic or curative agent for keratoconjunctive disorder - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To search a new pharmaceutical application of 2-phenyl-1,2-benzisoselenazol-3(2H)-one or salts thereof. <P>SOLUTION: The 2-phenyl-1,2-benzisoselenazol-3(2H)-one or salts thereof, because of exerting excellent prophylactic and ameliorative effects in keratoconjunctive disorder models, are useful as prophylactic or curative agents for keratoconjunctive disorders including dry eye, punctate cortical keratopathy, epithelium anterius corneae deletion, corneal erosion, corneal ulcer, tunica conjunctiva epitheloid deletion, keratoconjunctivitis sicca, superior limbic keratoconjunctivitis, filamentose karatoconjunctivitis, keratitis and conjunctivitis. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to dry eye, punctate superficial keratopathy, corneal epithelial defect, corneal erosion, corneal ulcer, comprising 2-phenyl-1,2-benzisoselenazol-3 (2H) -one or a salt thereof as an active ingredient, The present invention relates to a preventive or therapeutic agent for keratoconjunctival disorders such as conjunctival epithelial defect, dry keratoconjunctivitis, upper ring keratoconjunctivitis, filiform keratoconjunctivitis, keratitis, and conjunctivitis.

  The cornea is a transparent avascular tissue with a diameter of about 1 cm and a thickness of about 1 mm. The conjunctiva is a mucous membrane covering the surface of the eyeball behind the corneal margin and the back of the eyelid. It is known to have a significant effect on visual function. Keratoconjunctival disorders caused by various diseases such as corneal ulcer, keratitis, conjunctivitis, dry eye, etc., if repair is delayed for some reason or prolonged without repair, the tissue where the cornea and conjunctiva are connected Therefore, the normal construction of the epithelium is adversely affected, and even the structure and function of the corneal stroma and the endothelium may be impaired. In recent years, with the development of cell biology, factors involved in cell division, migration, adhesion, extension, differentiation, etc. have been elucidated, and these factors play an important role in the repair of corneal disorders. (Non-Patent Document 1, Non-Patent Document 2).

  On the other hand, Non-Patent Document 3 describes that 2-phenyl-1,2-benzisoselenazol-3 (2H) -one (generic name: ebselen, hereinafter referred to as “ebselen”) has antioxidant activity. Patent Document 1 describes that ebselen is effective as a therapeutic agent for cerebral arteriosclerosis and chronic cerebral circulation insufficiency.

However, there have been no reports on the pharmacological effects of these compounds on eye diseases such as keratoconjunctival disorders.
JP 2001-261555 A Eyesight, 46, 738-743 (1992) Ophthalmic Surgery, 5, 719-727 (1992) Proc. Natl. Acad. Sci. USA, 100 (13), 7919-7924 (2003)

  Searching for new medicinal uses for ebselen is an interesting challenge.

  The inventors of the present invention conducted intensive research in order to search for a new pharmaceutical use of the above compound. In a corneal disorder healing efficacy test using a corneal disorder model, the above compound or a salt thereof is excellent against corneal disorder. The present inventors have found that the present invention exerts preventive and improving effects, and has reached the present invention. A similar test was conducted for known compounds having an antioxidative effect in the same manner as ebselen, but ebselen exhibited a much higher effect than those known compounds, confirming the excellent usefulness of ebselen. It was.

  That is, the present invention comprises ebselen or a salt thereof as an active ingredient, dry eye, punctate superficial keratopathy, corneal epithelial defect, corneal erosion, corneal ulcer, conjunctival epithelial defect, dry keratoconjunctivitis, upper limbal keratoconjunctivitis, filamentous It is a preventive or therapeutic agent for keratoconjunctival disorders such as keratoconjunctivitis, keratitis, and conjunctivitis.

Ebselen of the present invention is a condensed heterocyclic compound represented by the following chemical structural formula [I].

  The salt of the above compound is not particularly limited as long as it is a pharmaceutically acceptable salt, such as a salt with an inorganic acid such as hydrochloric acid, nitric acid, sulfuric acid, acetic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, etc. Examples include salts with organic acids. In addition, the said compound may take the form of the solvate.

  In the present invention, the keratoconjunctive disorder refers to those in which the cornea or conjunctiva is damaged due to various factors such as tear abnormality, metabolic abnormality, external injury, etc., for example, dry eye, punctate superficial keratopathy Corneal epithelial defect, corneal erosion, corneal ulcer, conjunctival epithelial defect, dry keratoconjunctivitis, upper ring keratoconjunctivitis, filiform keratoconjunctivitis, keratitis, conjunctivitis and the like. Further, in the present invention, dry eye refers to lacrimation, dry eye, hypoxia, Sjogren's syndrome, dry keratoconjunctivitis, Stevens-Johnson syndrome, lacrimal gland dysfunction, meibomian gland dysfunction, blepharitis, VDT (Visual Display Terminal) Refers to keratoconjunctival disorders associated with work, surgery, drugs, trauma, contact lens wearing, etc., or symptoms associated with the keratoconjunctival disorders.

  The preventive or therapeutic agent for keratoconjunctival disorders of the present invention can be administered either orally or parenterally (instillation, transdermal, etc.). Examples of the dosage form include eye drops, eye ointments, skin ointments, injections, tablets, capsules, granules, fine granules, powders and the like. These can be prepared using commonly used techniques. For example, the eye drops are isotonic agents such as sodium chloride and concentrated glycerin, buffering agents such as sodium phosphate and sodium acetate, polyoxyethylene sorbitan monooleate, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil A surfactant such as sodium citrate and sodium edetate, and a preservative such as benzalkonium chloride and paraben can be used as necessary. The pH may be in the range acceptable for ophthalmic preparations, but is preferably in the range of 4-8.

  The eye ointment can be prepared using a commonly used base such as white petrolatum or liquid paraffin. In addition, oral preparations such as tablets, capsules, granules, fine granules and powders are bulking agents such as lactose, crystalline cellulose, starch and vegetable oil, lubricants such as magnesium stearate and talc, hydroxypropyl cellulose. -Binders such as polyvinylpyrrolidone, disintegrants such as carboxymethylcellulose calcium, low-substituted hydroxypropylmethylcellulose, coating agents such as hydroxypropylmethylcellulose, macrogol, silicone resin, A coating agent such as a gelatin film can be added as necessary.

  The dose of the above compound can be appropriately selected depending on symptoms, age, dosage form, etc., but in the case of eye drops, 0.000001 to 1% (w / v), preferably 0.0001 to 0.1% (w / v). Can be instilled once or several times a day. In the case of oral preparations, 0.1 to 5000 mg per day, preferably 1 to 1000 mg may be administered once or divided into several times.

  As will be described later, when the following pharmacological test was performed, ebselen exerts an excellent preventive and ameliorating effect in a corneal disorder model, so dry eye, punctate superficial keratopathy, corneal epithelial defect, corneal erosion, corneal ulcer It is useful as a preventive or therapeutic agent for keratoconjunctive disorders such as conjunctival epithelial defect, dry keratoconjunctivitis, upper limbal keratoconjunctivitis, filiform keratoconjunctivitis, keratitis, and conjunctivitis.

  The results of pharmacological tests and formulation examples are shown below, but these examples are for better understanding of the present invention and do not limit the scope of the present invention.

[Pharmacological test]
1. Curative efficacy test of corneal injury using rats with extraorbital lacrimal gland excision Using male SD rats, a corneal injury model was developed according to the method of Fujihara et al. (Invest. Ophthalmol. Vis. Sci 42 (1) 96-100 (2001)). Produced. After the creation of a corneal disorder model, a modified method (new ophthalmology, 21 (1), 87-90 (2004)) of the method of Miyata et al. (Ophthalmology Clinic, 48 (2), 183-188 (1994)) Then, the improvement rate of the corneal disorder was determined.

(experimental method)
Male SD rats were used for general anesthesia by administration of nembutal. Subsequently, the extraorbital lacrimal gland was removed and corneal injury was induced over 2 months.

Next, ebselen was administered as follows. Further, as a comparative compound, 3-methyl-1-phenyl-2-pyrazolin-5-one (hereinafter referred to as “edaravone”) represented by the following formula [II], and (3R) — represented by the following formula [III]: 1,2-dithiolane-3-pentanoic acid (hereinafter referred to as “α lipoic acid”) was administered as follows. Edaravone and α-lipoic acid are known to have antioxidant activity (J. Radiat. Res., 45, 319-323 (2004), J. Nutr., 133, 3327-3330 (2003)). As described above, ebselen is common to edaravone and α-lipoic acid in that ebselen has the same action.

Ebselen eye drops:
A physiological saline solution of ebselen (25 μM) was instilled into both eyes 6 times a day for 14 days (8 per group).

Edaravone eye drops:
A physiological saline solution of edaravone (200 μM) was instilled into both eyes 6 times a day for 14 days (8 per group).

Alpha lipoic acid eye drops:
A saline solution of α-lipoic acid (200 μM) was instilled into both eyes 6 times a day for 14 days (8 per group).

  In the control group, physiological saline was instilled into both eyes 6 times a day for 14 days (8 animals per group).

  14 days after the start of instillation, the damaged part of the cornea was stained with fluorescein. The degree of staining with fluorescein was scored according to the following criteria for each of the upper, middle, and lower parts of the cornea, and the improvement rate of corneal injury was calculated from the average value of the scores of the respective parts.

(Criteria)
0: Not dyed.

  1: Dyeing is sparse and each dot-like dyeing part is separated.

  2: The dyeing is moderate, and some of the dotted dyed portions are adjacent.

  3: Dyeing is dense and each dot-like dyeing part is adjacent.

  In addition, 0.5 was provided as an intermediate value between each score.

(result)
Improvement of the ebselen (25 μM) ophthalmic group, edaravone (200 μM) and α-lipoic acid (200 μM) ophthalmic group according to the following formula using the average score of the control group (saline) as a reference (improvement rate: 0%) The rate was calculated. This is shown in Table 1. The average score is the average of 8 cases in each group.

Improvement rate (%) = {(control) − (administered compound)} / degree of failure × 100
Disability level = (control)-(normal eye)

(Discussion)
As is apparent from the results of the above pharmacological tests using rats, ebselen shows a marked improvement rate compared to edaravone and α-lipoic acid. In particular, it is a surprising result that ebselen shows an improvement of more than 2 times over edaravone and alpha lipoic acid, even though it is administered at 1/8 times the concentration of edaravone and alpha lipoic acid. Therefore, it was shown that ebselen has a marked improvement effect on keratoconjunctival disorders.

2. Pharmacological test using SAMP10 According to the method of Hirai, Shigaki et al. (Japanese Patent Application No. 2006-104913), the effect of ebselen on corneal injury was investigated using Senescence-Accelerated Mouse P10 (hereinafter referred to as SAMP10). evaluated. As a control animal, SAMR1 showing normal aging was used.

(experimental method)
SAMP10 develops corneal damage with age. Therefore, corneal damage of female SAMP10 at the age of 16 weeks before aging change was evaluated, and then administration of ebselen solution or its base was started. The same evaluation was performed again at 24 weeks of age, 8 weeks after the start of administration. Similarly, the corneal injury of female SAMR1 at the age of 16 weeks was evaluated, and then the administration of the base was started, and the same evaluation was performed again at the age of 24 weeks, 8 weeks after the start of the administration.

(Method of administration)
6.67 μL of ebselen solution suspended in 1% (W / V) aqueous methylcellulose solution (prepared by dissolving methylcellulose in ultrapure water) at concentrations of 0.45 mg / mL and 4.5 mg / mL per mouse body weight In this ratio, the female SAMP10 was orally administered once a day to obtain a 3 mg / kg group and a 30 mg / kg group, respectively. Administration was carried out 5 days a week (Monday, Tuesday, Wednesday, Thursday, Friday) for 8 weeks. As a control, a 1% (W / V) methylcellulose aqueous solution was orally administered to SAMP10 in the same manner at a rate of 6.67 μL / g body weight of the mouse. Moreover, 1% (W / V) methylcellulose aqueous solution was orally administered similarly to female SAMR1 which is a control animal. The number of examples of each group of SAMP10 is 22 eyes (11 animals), and the number of examples of SAMR1 is 16 eyes (8 animals).

(Evaluation methods)
The corneal lesions of SAMP10 and SAMR1 immediately before the start of administration and 8 weeks after the start of administration were stained with fluorescein. For each of the upper, middle and lower parts of the cornea, the degree of staining with fluorescein was scored based on the criteria described in “1. Curative efficacy test for corneal injury using an extraorbital lacrimal gland excision rat model”. The score of the corneal injury was scored (total of 9 points).

(result)
The fluorescein staining scores for SAMP10 and SAMR1 at 16 and 24 weeks of age are shown in Table 2. In addition, the score in a table | surface is the average +/- standard error in each group.

^## p <0.01 Comparison with SAMR1 (Student's t test)
* p <0.05 Comparison with SAMP10 base administration group (Dunnett's test)
(Discussion)
As is apparent from Table 2, at the age of 16 weeks, there was no significant difference between the SMR1 base administration group and the SAMP10 base administration group (Student's t-test). There was no significant difference (Tukey test). At 24 weeks of age, the SAMP10 base administration group was found to have a significantly increased staining score compared to the SAMR1 base administration group. That is, it was confirmed that SAMP10 develops a corneal disorder with aging. Under these conditions, it was observed that both the 3 mg / kg and 30 mg / kg ebselen administration groups had significantly lower staining scores than the SAMP10 base administration group. That is, ebselen was found to have a preventive and ameliorating effect on corneal disorders.

[Formulation example]
A typical formulation example using ebselen is shown below.

Formulation Example 1 Eyedrops 100mg Ebselen 10mg
Sodium chloride 900mg
Sterilized purified water appropriate amount Concentration 0.001% (w / v), 0.03% (w / v), 0.1% (w / v), 0.3% (w) by changing the amount of ebselen added / V), 1.0% (w / v) eye drops.

Formulation Example 2 Eyedrops 100ml Ebselen 100mg
Sodium chloride 800mg
Disodium hydrogen phosphate 100mg
Sodium dihydrogen phosphate Appropriate amount Sterilized purified water Appropriate amount By changing the amount of ebselen added, concentrations of 0.05% (w / v), 0.3% (w / v), 0.5% (w / v), A 1% (w / v) eye drop can be prepared.

Formulation Example 3 Ebselen 0.3g in 100g of eye ointment
Liquid paraffin 10.0g
White petrolatum appropriate amount By changing the amount of ebselen added, an eye ointment having a concentration of 1% (w / w) and 3% (w / w) can be prepared.

Formulation Example 4 Ebselen 1mg in 100mg tablet
Lactose 66.4mg
Corn starch 20mg
Carboxymethylcellulose calcium 6mg
Hydroxypropylcellulose 6mg
Magnesium stearate 0.6mg
Ebselen and lactose are mixed in a mixer, carboxymethylcellulose calcium and hydroxypropylcellulose are added thereto, this mixture is granulated, the resulting granules are dried and sized, and magnesium stearate is added to the sized granules. In addition, the mixture is mixed, and the mixture is tableted with a tableting machine. Moreover, the tablet whose content in 100 mg is 0.1 mg, 10 mg, and 50 mg can be prepared by changing the addition amount of ebselen.

Claims (4)

A prophylactic or therapeutic agent for keratoconjunctival disorder comprising 2-phenyl-1,2-benzisoselenazol-3 (2H) -one or a salt thereof as an active ingredient. The keratoconjunctival disorder is dry eye, punctate superficial keratopathy, corneal epithelial defect, corneal erosion, corneal ulcer, conjunctival epithelial defect, dry keratoconjunctivitis, keratoconjunctivitis keratoconjunctivitis, keratoconjunctivitis, keratitis or conjunctivitis 1. The preventive or therapeutic agent according to 1. The prophylactic or therapeutic agent according to claim 1, wherein the administration form is ophthalmic administration or oral administration. The preventive or therapeutic agent according to claim 1, wherein the dosage form is an eye drop, an eye ointment, a tablet, a fine granule or a capsule.