JP2004059432A - Therapeutic agent for corneopathy - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a therapeutic agent for corneopathy effective especially for treatment of ectocorneopathy produced from various causes by exhibiting selective proliferation promoting actions on ectocorneal cells. <P>SOLUTION: The therapeutic agent for the corneopathy comprises α-tocopheryl retinoate exhibiting the selective proliferation promoting actions on the ectocorneal cells. <P>COPYRIGHT: (C)2004,JPO

Description

【００１９】
実施例９：
トレチノイントコフェリル０．１ｇをとり、流動パラフィンで研和泥状とした後、眼科用白色ワセリンを徐々に加えて練合し、全質均等として全量１００ｇとした。得られた眼軟膏を、滅菌した軟膏チューブに充填した。以上の操作は無菌環境下で無菌的に行った。
【００２０】
【発明の効果】
本発明によれば、角膜上皮細胞に対して選択的な増殖促進作用を発揮させることができることにより、種々の原因によって引き起こされる角膜上皮障害の治療にとりわけ有効な角膜障害治療剤が提供される。
【図面の簡単な説明】
【図１】実施例におけるトレチノイントコフェリルの角膜上皮細胞に対する作用を示すグラフである。
【図２】実施例におけるトレチノイントコフェリルの角膜実質細胞に対する作用を示すグラフである。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a therapeutic agent particularly effective for a corneal disorder accompanied by a corneal epithelial disorder caused by various causes.
[0002]
[Prior art]
The cornea is an avascular tissue located at the forefront of the eye, and plays an important role of transmitting external light while maintaining transparency. It also acts as a barrier, preventing external stimuli and the invasion of pathogens. The structure of the cornea is roughly divided into, in order from the outside, a corneal epithelial layer (consisting of 5 to 7 layers of epithelial cells), Bowman's membrane (basement membrane of the corneal epithelial layer), and a corneal stromal layer (stromal cells and collagen fibrous plate) , A corneal endothelial layer (consisting of one layer of endothelial cells), a Descemet's membrane (basement membrane of endothelial cells), and a corneal endothelial layer.
[0003]
For example, diseases such as corneal herpes, bacterial and fungal keratitis, spring catarrh, Sjogren's syndrome, Steven-Johnson syndrome, dry eye syndrome (dry eye), vitamin A or B2 deficiency may be accompanied by corneal disorders. In addition, the cornea may be damaged by external stimuli such as burns, trauma, chemical substances such as alkalis and acids, drugs, ultraviolet rays, wearing of contact lenses, foreign substances, eyelashes, and drying out of the air. Further, the cornea may be damaged by surgical invasion of the cornea such as corneal refractive surgery, cataract surgery, intraocular lens insertion surgery, corneal transplantation surgery, corneal foreign body removal surgery, glaucoma surgery, and the like.
Among them, when the corneal epithelium is damaged due to damage, in the healing process, the epithelial cells around the damaged part need to compensate for the deficient portion of the epithelial cells by expanding and moving with proliferation. Promoting the proliferation of corneal epithelial cells promotes cell growth and migration and also promotes repair of the corneal epithelial layer, and is therefore important for promptly restoring the barrier function of the cornea.
Conventionally, as a treatment method for corneal disorders due to various causes, particularly corneal epithelial disorders, hyaluronic acid ophthalmic treatment has been applied with the expectation of an action of promoting the progress of corneal epithelial cells. In recent years, treatment methods such as fibronectin ophthalmic therapy, which focuses on the promoting action of corneal epithelial cells, and epidermal growth factor (EGF) epithelial growth factor (EGF), which is expected to promote growth, have been studied.
[0004]
[Problems to be solved by the invention]
However, none of the above therapies has always produced satisfactory results, and there are many problems to be solved in terms of side effects. For example, EGF ophthalmic therapy is characterized by treating corneal disorders by promoting the proliferation of corneal epithelial cells, but has problems in angiogenesis and carcinogenesis. Furthermore, EGF has a problem that it is inactivated by benzalkonium chloride, which is a general preservative of eye drops, and has many practical problems. Therefore, a safe and highly effective therapeutic agent for corneal disorders is desired.
In addition, the causes of corneal disorders are diverse and their symptoms are also various. Therefore, when treating corneal disorders, it is essential to clarify the target tissue and cells and select and use the appropriate therapeutic agent. It can be said to be a desirable figure. However, it must be said that therapeutic agents capable of performing such treatment have not been practically used. For example, EGF eye drop therapy is not appropriate to apply this therapy depending on the disorder of the cornea, because EGF has a growth promoting effect not only on epithelial cells constituting the cornea but also on parenchymal cells and endothelial cells. In some cases. More specifically, when only the corneal epithelial layer is damaged, if EGF ophthalmic treatment is applied, EGF also promotes the proliferation of corneal parenchymal cells that do not need to act and causes corneal opacity. There is a risk of lowering the patient's QOL (Quality of Life). In view of the above, since the corneal epithelial layer is susceptible to damage, only the corneal epithelial layer often suffers from the damage. I have. Further, even when the corneal disorder extends to the corneal stroma, since a rapid proliferation of corneal epithelial cells is effective for the recovery of corneal tissue, a therapeutic agent that promotes the proliferation of corneal epithelial cells is highly valuable. Things.
Therefore, an object of the present invention is to provide a therapeutic agent for corneal disorders that is particularly effective for treating corneal epithelial disorders caused by various causes, by being able to exert a selective proliferation promoting action on corneal epithelial cells. And
[0005]
[Means for Solving the Problems]
The present inventors have conducted various studies in view of the above points, and as a result, an ester of α-tocopherol and retinoic acid (vitamin A acid), which is known as an active ingredient of a therapeutic agent for skin diseases. With respect to α-tocopheryl retinoate, it has been found that it is an effective compound for treating corneal disorders, and an interesting finding that it can exert a selective growth promoting action on corneal epithelial cells.
[0006]
The present invention has been made based on the above findings, and the therapeutic agent for corneal disorders of the present invention contains α-tocopheryl retinoate as described in claim 1.
Further, the therapeutic agent for corneal epithelial disorder of the present invention contains α-tocopheryl retinoate as described in claim 2.
Further, the corneal epithelial cell growth promoting agent of the present invention contains α-tocopheryl retinoate as described in claim 3.
[0007]
BEST MODE FOR CARRYING OUT THE INVENTION
The therapeutic agent for corneal disorder of the present invention is characterized by containing α-tocopheryl retinoate. According to the present invention, a therapeutic agent for corneal epithelial disorder based on a selective corneal epithelial cell growth promoting action, comprising α-tocopheryl retinoate, which has been confirmed to be safe and has excellent chemical stability, as an active ingredient. Can be provided.
With respect to the use of α-tocopheryl retinoate in the ophthalmic field, WO 00/21528 proposes an ophthalmic pharmaceutical composition using α-tocopheryl retinoate in combination with vitamin A. It states that it can protect eyes from ozone and ozone. However, the effect of α-tocopheryl retinoate in relation to the site of the cornea has not been studied at all, and the effect cannot be recalled from the above description. Am. J. Ophthalmol. 95: 353-358 (1983) states that vitamin A acid has an effect of promoting healing of experimental corneal epithelial disorder, but has no effect on vitamin A, vitamin A palmitate or acetate. It has been. Also, Ann. Ophytal. 19: 175-180 (1987) states that vitamin A ointment enhances healing of cataract incisions, but vitamin A has poor chemical stability. Therefore, based on such known findings, α-tocopheryl retinoate is a compound effective for the treatment of corneal disorders, and α-tocopheryl retinoate is selectively promoted for corneal epithelial cells. It is quite surprising that it can work.
[0008]
Α-Tocopheryl retinoate, which is an active ingredient of the therapeutic agent for corneal disorder of the present invention, is an ester of α-tocopherol and retinoic acid as described above. α-Tocopherol includes DL-α-tocopherol, D-α-tocopherol and L-α-tocopherol, and retinoic acid includes all-trans-retinoic acid, 13-cis-retinoic acid, 11-cis- Retinoic acid, 9-cis-retinoic acid. Preferred retinoic acids are all-trans-retinoic acid and 13-cis-retinoic acid. Particularly preferable α-tocopheryl retinoate includes an ester of DL-α-tocopherol and all-trans-retinoic acid (tretinoin tocopheryl: Merck Index 12th).  edition, no. 9639).
[0009]
The therapeutic agent for corneal disorders of the present invention is provided in various preparation forms such as eye drops, eye ointments and gels. Among them, a desirable formulation form includes eye drops. These preparations, together with the active ingredient α-tocopheryl retinoate, are commonly used in preparing individual preparations such as buffers, dissolution aids, isotonic agents, stabilizers, preservatives, The composition comprises an ophthalmic pharmaceutical composition in which various additives such as an agent, a chelating agent, a pH adjuster, and a cooling agent are blended in a usual amount. Explaining the additives specifically, examples of the buffer include boric acid or a salt thereof (borax or the like), citric acid or a salt thereof (sodium citrate or the like), phosphoric acid or a salt thereof (sodium monohydrogen phosphate) )), Tartaric acid or a salt thereof (such as sodium tartrate), gluconic acid or a salt thereof (such as sodium gluconate), acetic acid or a salt thereof (such as sodium acetate), various amino acids, or a combination thereof. Examples of the dissolution aid include polyoxyethylene hydrogenated castor oil (polyoxyethylene hydrogenated castor oil 60, etc.), polyethylene glycol (macrogol 4000, etc.), polyoxyethylene sorbitan higher fatty acid ester (polysorbate 80, etc.), polyoxyethylene (POE) -polyoxypropylene (POP) block copolymer (poloxamer 407 etc.), propylene glycol and the like. Examples of the tonicity agent include inorganic salts (eg, sodium chloride, potassium chloride, calcium chloride), sugars (eg, mannitol, glucose), and polyhydric alcohols (eg, glycerin, propylene glycol). Examples of the stabilizer include sodium edetate, cyclodextrin, sulfite, citric acid or a salt thereof, butylhydroxytoluene, and the like. As a preservative, benzalkonium chloride, benzalkonium chloride having 12 carbon atoms in the alkyl group, benzethonium chloride, chlorhexidine gluconate, chlorobutanol, sorbic acid, potassium sorbate, methyl paraoxybenzoate, methyl paraoxybenzoate, Examples thereof include propyl paraoxybenzoate, butyl paraoxybenzoate, polyquartenium-1 (trade name: Polyquad), polyaminopropybiguanide (trade name: Cosmosil CQ), alkylaminoethyl glycine hydrochloride, cetylpyridinium chloride, thimerosal, and the like. Examples of the thickener include polyethylene glycol, polyvinyl alcohol, polyvinyl pyrrolidone, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, sodium chondroitin sulfate, sodium hyaluronate, carboxymethyl cellulose, carboxyvinyl polymer and the like. Among them, hydroxyethylcellulose, hydroxypropylmethylcellulose, and sodium chondroitin sulfate are desirable viscous agents because they contribute to moisturizing the cornea and improving the retention of drugs. Examples of the chelating agent include sodium edetate, sodium citrate and the like. Examples of the pH adjusting agent include hydrochloric acid, citric acid or a salt thereof, boric acid or a salt thereof, phosphoric acid or a salt thereof, acetic acid or a salt thereof, tartaric acid or a salt thereof, sodium hydroxide, potassium hydroxide, sodium carbonate, and carbonic acid. Sodium hydrogen and the like. Examples of the cooling agent include menthol, borneol, camphor, geraniol, limonene, eugenol, peppermint oil, eucalyptus oil, fennel oil, bergamot oil and the like. Further, it is desirable to add an antioxidant such as tocopherol acetate or sodium sulfite in consideration of the chemical stability of α-tocopheryl retinoate. Solvents that can be used when producing eye drops include sterilized purified water, purified water, physiological saline, and other suitable aqueous solvents. Examples of the non-aqueous solution used for the oily eye drops include vegetable oils (olive oil, soybean oil, sesame oil, cottonseed oil, etc.), liquid paraffin, mineral oil, propylene glycol, p-octyldodecanol, and the like. Examples of bases for ophthalmic ointments include oily bases (Vaseline, liquid paraffin, xylene 50W, plastibase, etc.), emulsion bases, water-soluble bases (hydroxypropylmethylcellulose, carboxymethylcellulose, macrogol) and the like. In addition to the above components, carrageenan, pectin, guaia gum, duran gum, agar, sodium alginate, albumin, curdlan, xanthan gum, gelatin and the like can be added to the gel. The pH of the eye drop in the case where the therapeutic agent for corneal disorder of the present invention is provided as an eye drop is not particularly limited as long as it is ophthalmically acceptable, and is usually in the range of pH 4 to 9, preferably pH 5 to 5. 8 range.
[0010]
The desired preparation is prepared by adding the above-mentioned buffering agent, dissolution aid, isotonic agent, stabilizer, preservative, thickener, chelating agent, pH adjusting agent to α-tocopheryl retinoate as an active ingredient. , A cooling agent and the like, and other components are appropriately blended to prepare an ophthalmic pharmaceutical composition in a usual manner, which is subjected to aseptic filtration under a sterile environment, and filled into a suitable sterile container to produce the composition. For non-filterable preparations such as eye ointments, operate aseptically in a sterile environment. The sterile container may be any of those usually used in the art depending on the form of the preparation, but in view of ensuring the stability of the preparation, for example, Teflon, silicon, polypropylene, polystyrene, polyethylene, polyethylene terephthalate It is desirable to use a container made of a material such as polyethylene naphthalate and polyarylate. In addition, for the purpose of ensuring higher formulation stability, the prepared ophthalmic pharmaceutical composition is filled in a suitable sterile container and closed, and then sealed together with a deoxidizer in a substantially oxygen-impermeable packaging. May be. As a package that is substantially impermeable to oxygen, a bag-like or tubular-like package composed of a film in which a synthetic resin film such as polyethylene, polypropylene, polyvinyl alcohol, polyvinylidene chloride, or the like is laminated on an aluminum foil is used. However, any sealable material made of a material that does not substantially transmit oxygen can be used for convenience.
[0011]
The amount of α-tocopheryl retinoate in the therapeutic agent for corneal disorder of the present invention is usually 0.00001 to 10 parts by weight, preferably 0.00005 to 5 parts by weight, more preferably 0.00005 to 5 parts by weight, per formulation. 0.0001 parts by weight to 1 part by weight. For the application, it may be instilled or applied once to several times a day. The amount used is determined according to the degree of symptoms and the size of the affected area.
[0012]
The therapeutic agent for corneal disorders of the present invention may contain various pharmaceutically active ingredients as needed, for example, decongestants (naphazoline hydrochloride, tetrahydrozoline hydrochloride, phenylephrine hydrochloride, etc.), neostigmine methyl sulfate, anti-inflammatory and astringent (epsilon-aminocapron) Acid, allantoin, berberine chloride, zinc sulfate, lysozyme chloride, etc., antihistamines (diphenhydramine hydrochloride, isotipendyl hydrochloride, chlorpheniramine maleate, etc.), fat-soluble vitamins [active vitamin E, vitamin A], water-soluble vitamins [ Activated vitamin B2, vitamin B6, vitamin B12], amino acids (potassium L-aspartate, magnesium L-aspartate, aminoethylsulfonic acid, sodium chondroitin sulfate, etc.), sulfa drugs, fungicides (sulfur, isopropylmethyl) Phenol, hinokitiol, etc.), local anesthetics (lidocaine, lidocaine hydrochloride, procaine hydrochloride, dibucaine hydrochloride, etc.), can be appropriately compounded sugars (glucose, etc.) and the like.
[0013]
【Example】
The present invention will be described in more detail by the following experiments and examples, but the present invention is not limited thereto.
[0014]
Experiment 1: Action of Tretinoin Tocopheryl on Corneal Epithelial Cells Normal rabbit corneal epithelial cells (NRCE) were seeded at 2 × 10 ⁴ cells / well on a 24-well culture plate, and a medium for rabbit corneal epithelial cells was used as a culture medium. (RCGM: manufactured by Kurabo Industries), and cultured at 37 ° C. under 5% CO ₂ . Twenty-four hours later, the culture medium is removed together with the non-adherent cells, and a culture medium containing 10 ⁻⁸ , 10 ⁻⁷ , 10 ^−6, and 10 ⁻⁵ M tretinoin tocopheryl is added to each well, and the cells are further cultured for 72 hours. did. After completion of the culture, the culture solution containing tretinointocopheryl was removed, a new culture solution was added to each well, and the number of cells was measured using Cell Counting Kit-8 (manufactured by Dojindo Chemical). The results are shown in FIG.
[0015]
Experiment 2: Effect of tretinoin tocopheryl on corneal stromal cells A corneal stromal section from which the epithelium and endothelial tissue had been removed from the cornea extracted from a Japanese white rabbit was subjected to 1000 U / ml dispase (manufactured by Godo Shusei). Enzyme treatment was performed at 37 ° C. for 60 minutes to isolate keratocytes. On a 24-well culture plate, isolated corneal stromal cells were seeded at 2 × 10 ⁴ cells / well, and Eagle MEM medium (manufactured by Nissui Pharmaceutical Co., Ltd.) supplemented with 2% fetal bovine serum (manufactured by Invitrogen) as a culture solution ) And cultured at 37 ° C. under 5% CO ₂ . Twenty-four hours later, the culture medium is removed together with the non-adherent cells, and then containing ^10-8 , ^10-7 , ^10-6, and ^10-5 M tretinoin tocopheryl or 10 ng / ml EGF (manufactured by R & D). The culture solution was added to each well, and the cells were further cultured for 72 hours. After completion of the culture, the culture solution containing tretinointocopheryl or EGF was removed, and a new culture solution was added to each well, and the number of cells was measured in the same manner as in Experiment 1. FIG. 2 shows the results.
[0016]
Experimental result:
As is clear from FIGS. 1 and 2, tretinoin tocopheryl showed a corneal epithelial cell growth promoting action in a concentration-dependent manner at a solution concentration of 10 ⁻⁸ M or more. On the other hand, tretinoin tocopheryl did not show a growth promoting effect on corneal stromal cells even at a solution concentration of 10 ⁻⁸ M or more. From the above results, tretinoin tocopheryl was found to be useful for the treatment of corneal disorders associated with corneal epithelial disorders because it can exert a selective growth promoting action on corneal epithelial cells.
[0017]
Examples 1 to 8:
The components of the formulation shown in Table 1 were dissolved in purified water, and if necessary, 0.1 N hydrochloric acid and 0.1 N sodium hydroxide were added in appropriate amounts to adjust to a predetermined pH, and the total amount was adjusted to 100 ml. The obtained aqueous solution was sterilized by filtration under a sterile environment, and filled in a container of eye drops which had been washed and sterilized to obtain a colorless and clear eye drop.
[0018]
[Table 1]

[0019]
Example 9:
After taking 0.1 g of tretinoin tocopheryl and making it into a muddy state with liquid paraffin, white petrolatum for ophthalmology was gradually added and kneaded to make a total amount of 100 g as a uniform material. The obtained eye ointment was filled in a sterilized ointment tube. The above operation was performed aseptically in a sterile environment.
[0020]
【The invention's effect】
According to the present invention, a therapeutic agent for corneal disorders which is particularly effective for treating corneal epithelial disorders caused by various causes is provided by being able to exert a selective proliferation promoting action on corneal epithelial cells.
[Brief description of the drawings]
FIG. 1 is a graph showing the effect of tretinoin tocopheryl on corneal epithelial cells in Examples.
FIG. 2 is a graph showing the effect of tretinoin tocopheryl on corneal stromal cells in Examples.

Claims (3)

A therapeutic agent for corneal disorders, which comprises α-tocopheryl retinoate. A therapeutic agent for corneal epithelial disorder, comprising α-tocopheryl retinoate. A corneal epithelial cell proliferation promoter comprising α-tocopheryl retinoate.

Images