WO2001091779A1 - Remedies for corneal injuries - Google Patents

Abstract

Drugs to be used for preventing and treating corneal injuries, in particular, physical, chemical and mechanical injuries and endogenously induced failures in the cornea including subepithelial basement membrane located between ectocornea and endothelium corneae, Bowman's membrane, keratocytes, matrix among collagen fibers and Descemet's membrane. Remedies for corneal injuries having an effect of restoring the altered proteoglycan composition of keratocytes into the normal state, which contain urinastatin as the active ingredient.

Description

 Specification

 Corneal wound treatment

Technical field

 The present invention relates to corneal damage, in particular, physical, chemical, and mechanical damage to the cornea, including the subepithelial basement membrane, Bowman's membrane, corneal stroma, collagen fibril matrices, and Descemet's membrane present between the corneal epithelium and the corneal endothelium. And a drug used for the prevention and treatment of intrinsic corneal injury. Background art

 The cornea is an anterior pole of the eyeball, transparently covers the anterior chamber of the lens, and is a tissue having an orderly layer structure composed of an epithelial cell layer, a Bomman membrane, a stromal layer, a Descemet's membrane, and an endothelial cell layer.

 The corneal stroma is mainly composed of collagen and proteoglycans. Proteodarican is a mucopolysaccharide-protein complex, a compound in which daricosaminodarican (GAG) and a core protein are covalently linked.

The proteodalicans that make up the corneal stroma are mainly Lumican, where GAG is keratan sulfate, and Decorin, where GAG is chondroitin Z dermatan sulfate.The length of the chondroitin / dermatan sulfate side chain is the length of collagen fibrils. GAG corresponds to the distance between centers, and the length of the keratan sulfate side chain corresponds to the length of the gap between collagen fibrils. S cott, JE r P roteoglycan— fibri 1 1 ercollagen interaction, J., 2 52, 31 3-323, 1988, S cott, J. E .: Mo rphometryofc up r ome r on icblue— stainedproteoglycan mo leculesinani ma lcorneas, versusthatofpurifiedp roteoglycanssainedinv itro, imp liesthttertiarystruct urescontributetocorne a 1 ultrastructure., J. An at., 180, 155-1 64, 1 9 9 2), horns! ^ Building a real three-dimensional lattice-like structure (Benedek , GB: Theory of transparency of the eye., Ap pl. Opt. 10, 459—473, 1971) has been proposed by Scott et al. Among the GAGs that make up the cornea, keratan ^ acid is particularly important, and ^^ mandarin, whose GAG is keratan sulfate, accounts for 50-70% of the total corneal osteoglycans. Lumican plays an important role in the normal corneal stroma, such as the arrangement of collagen fibers and the adjustment of the interfiber space. When the cornea or corneal stroma is injured for any reason, the parenchymal lumican is depleted and the majority of decorin is larger than normal. Such changes in proteoglycin force increase the distance between collagen fibers ゃ the diameter of collagen fibrils, and the distance between collagen fibers and the diameter of collagen fibrils increase the wavelength of light. (200 nm to 250 nm), light is scattered in the cornea and causes corneal (parenchyma) opacity (A ssi 1, KK, eta 1 .:

Wo u n d h e a l i n g i n r e s p o n se t o k e r a t o r e f e a c t i v e s u r g e r y., S u r v. Op h t h a l mo l., 3

8, 28 9-30, 19 9 3).

 As healing of the corneal or corneal wound progresses, lumican eventually increases to its original level, the size and composition of each proteodalican normalizes, and collagen fibrils are reconstructed (Assil, KK: S ur v. O phthal mo l., 3 8, 2

8 9-3 0 2, 1 9 9 3).

In addition, when the cornea or the corneal stroma is wounded, the molecular weight of the proteoglycan in the parenchyma also changes.This includes changes in the position of GAG sulfate and the change in the water content due to changes in the proportion of GAG species. It is said that the increase has an effect (H ass _e 11, J.R., eta 1.: P roteoglycanchangesdur ingrestoration of trans parencyincornealscars., Arc h. Biom em. Biophys., 222, 3 6—

369, 198 3). In addition, in patients with mottled corneal dystrophy, many proteoglycans containing keratan in the cornea lack a sulfate group (Miura, RJ, etal .: Proteogrycanbiosynthes isbyhumancorneasfrom atientwithtye 1 and 2ma cularcornea). 1 dystroph y., J. Biol. Chem.'2 65 (26), 15 947—15 955, 1 990). This is attributed to a deficiency in the gene for sulfotransferase into keratan.

 In order to maintain normal translucency, it is necessary that the ratio of lumican with keratan sulfate as GAG is high. However, deficiency of sulfotransferase produces lumican and other proteoglycans, which are keratan-free mucopolysaccharides that have no sulfate group, and their solubility is extremely low, causing precipitation in the cornea and causing visual impairment. (E dwa rd, DP, eta 1 .: Ma cu 1 ardystrop ft. Yofthecorne a. A syst em icdisorderofkeratansu lfate me tabolis m., O phthal mo 1 ogy, 9 7 (9), 1 194-1200, 1 990). In this way, various proteoglycans and various GAGs that form them play a very important role in forming a regular and orderly lattice-like structure of the corneal stroma, and if the cornea is damaged for any reason, Cornea The type and composition of GAG and proteodalican change significantly compared to normal corneas. It is being elucidated that the type and composition ratio of the corneal healing approaches that of the normal cornea. Summary of the Invention

 In view of the above situation, the present invention provides a method for physically treating a cornea, particularly including a subepithelial basement membrane, a Bowman's membrane, a corneal stroma, a matrix between collagen fibers, and a Descemet's membrane existing between the corneal epithelium and the corneal endothelium. The purpose of the present invention is to provide a drug used for the prevention and treatment of chemical, mechanical damage and intrinsic corneal injury.

The present invention is intended to restore the altered composition of corneal stroma proteoglycin to a normal state. A therapeutic agent for corneal wounds, comprising perinastatin as an active ingredient.

 The present invention is also an agent for treating a corneal stromal wound, which comprises perinastatin as an active ingredient. Among them, those having an effect of returning the composition of the changed proteoglycan of the corneal stroma to a normal state are preferable.

 In addition, in this specification, returning the composition of corneal stromal proteodalican to a normal state also includes promoting the return of the corneal stromal proteoglycin composition to a normal state. Detailed Disclosure of the Invention

 Hereinafter, the present invention will be described in detail.

 As described above, the present inventors have found that if the cornea is damaged for any reason, the composition of proteodalican in the corneal substance changes, and the composition of proteogliin becomes normal as the corneal wound heals. By returning, we thought that a drug that restores the composition of proteodalican or GAG that changed due to a wound on the cornea to a normal corneal state or a drug that promotes it could be an effective therapeutic agent for corneal wounds. As a result of research, they have found that perinastatin has an effect of promoting the restoration of the composition of proteodalican, which has been changed, to normal, and completed the present invention. "

 As described above, in the normal corneal stroma, 50 to 70% of the proteodalican in the corneal stroma is occupied by Lumican, which is keratan sulfate, and GAG is Chondroitin / Dermatanic acid / Remican, by about 30%. Occupy. In addition, proteoglycans in which GAG is hyaluronic acid are also found. These proteoglycans exist with a substantially constant composition in normal corneal stroma, although there are individual differences.

However, if the corneal stroma is injured for any reason, the wound cornea depletes the lumican of proteoglycan (PG), the main component of the parenchymal lattice structure that maintains the transparency of the cornea, and decorin It is rapidly expressed and synthesized, and decorin is changed for Lumican. This increases the distance between the collagen fibers divided by the diameter of the collagen fibrils, ruptures the lattice structure, cloudes the corneal stroma, and loses corneal transparency. That is, in the healing process of the corneal wound, the amount ratio of Lumican Z decorin is Extremely fluctuates from about 73 to 0 ~ 2Zl Q ~ 8, and with the reconstruction of the corneal tissue, this PG content ratio will return to its original value, and the size and composition ratio of each PG will be normalized and collagen fibrils Is reconstructed (Assil, KK: Surv. Ophtha lmol., 38, 289—302, 1993), and is thought to restore corneal transparency. However, if this ratio is not restored and the amount of decorin is dominant, corneal remodeling will not improve and will eventually lead to chronic corneal opacity and scarring of the corneal epithelium.

 Furthermore, ECM is regarded as important as a molecular mechanism that regulates aqueous humor outflow resistance in the aqueous humor channel. ECM is localized in the trabecular meshwork endothelium, which is the main site of aqueous humor outflow resistance, and is considered to be a regulator of aqueous humor outflow resistance. Qualitative and quantitative abnormalities of the ECM component have been reported in glaucoma eyes.In particular, the overexpression of decorin gene in trabecular meshwork cells indicates abnormal expression of decorin in the trabecular meshwork endothelium- Biosynthesis enhances aqueous humor outflow resistance and is thought to be the etiology of glaucoma.

 As described above, decorin is a PG that plays an important role in corneal wound healing and ocular physiology, but even a slight increase in its physiological amount may cause serious eye disease. . Therefore, suppressing the overexpression of decorin in the cornea and trabecular meshwork is thought to lead to the treatment and prevention of serious eye diseases such as corneal opacity, corneal epithelial scarring and glaucoma. Therefore, in order to develop a therapeutic agent for corneal wounds in diabetic patients, which has been a major problem in the ophthalmology field because of its extremely intractable nature, we investigated changes in corneal GAG levels during the process of healing corneal wounds in diabetic rats. saw. As a result, the present inventors have found for the first time that in diabetic corneas, the amount of keratan sulfate reflecting the amount of lumican decreases and the amount of dermatan sulfate reflecting the amount of decorin increases in normal corneas. Was.

 The above-mentioned pelinastatin has an action of promoting the process of normalizing the composition of PG, and the UTI instillation has a normalizing effect on the abnormal change of the corneal GAG in the corneal wound healing process of the diabetic rat. This has been clarified by the present inventors' research.

Perinastatin is a glycoprotein with a molecular weight of about 67000 (measured by gel filtration chromatography) or a molecular weight of about 34000 (measured by SDS-polyacrylamide gel electrophoresis), of which about 35% is composed of sugar moieties. It is a well-known substance (Medical and Pharmaceutical Science, Vol. 33, No. 5, 108-109, 1995). Linastatin is marketed by Mochida Pharmaceutical under the trade name Miracrid.

 Perinastatin can be obtained from healthy male fresh urine by a general purification method, for example, by adsorbing on kieselguhr and silica gel, and then sequentially combining and purifying an ion exchange resin, gel filtration and the like.

 In recent years, due to the development of analytical methods, it has been known that perinastatin is composed of several types of isomers. The difference between these isomers is characterized by the degree of sulfonation of their constituent sugar moieties. It is considered that there is no difference in other points (antitrypsin activity, amino acid composition, N-terminal amino acid sequence, C-terminal amino acid sequence, sialic acid content, and peronic acid content) (Yuki, Y ., Eta 1 .: Biochimicaet Biophysica Acta, 1203, 298-303, 1993). It is clear that any of these isomers of perinastatin are included in perinastatin used in the present invention.

 The therapeutic agent for corneal wound of the present invention can be produced by mixing the above perinastatin with a pharmacologically acceptable additive.

 When the above pelinastatin is used as an active ingredient of the therapeutic agent for corneal wounds of the present invention, the concentration range of pelinastatin is not particularly limited, but 0.05 to 10,000 g / mL promotes normalization of the composition of PG in human corneal parenchyma. Therefore, it is preferable to use the same concentration range. By limiting the further concentration range to 0.5 to 5000 / igZmL, it is recognized that the normalization of the PG composition of the human cornea is significantly promoted, and it is more preferable to use the same concentration range.

The agent for treating a corneal wound of the present invention is preferably in the form of a topical administration agent, particularly an ophthalmic solution, an ointment, or a lyophilizer, but is not limited thereto. When used as eye drops or eye ointments, additives such as preservatives such as sodium dehydroacetate and methyl para-hydroxybenzoate, tonicity agents such as sodium chloride and glycerin, lipoxymethylcellulose, and polybutyl alcohol General substances such as a thickener, sodium edetate, and a stabilizer such as polysaccharide can be used, but are not limited thereto, and may be any range as long as they are ophthalmologically acceptable. Buffer added to the above various dosage forms It is preferable to use an isotonic solution that is isotonic and has a pH of 4 to 9, especially 5.5 to 8.0.

 In addition, the therapeutic agent for corneal wounds of the present invention contains fibronectin, hyaluronic acid and various glycosaminoglycans (chondroitin sulfate, dermatan sulfate, heparan sulfate, heparin, heparin, etc.) as long as the function of perinastatin as an active ingredient is not hindered. It can be used in combination with ophthalmic physiologically active substances such as rattan sulfate) and can be used as a powerful therapeutic agent for corneal wounds. Furthermore, it is also possible to mix and use steroidal anti-inflammatory drugs such as antibiotics and dalcocorticoid ゃ nonsteroidal anti-inflammatory drugs.

 The usage and dosage of the therapeutic agent for corneal wounds of the present invention vary depending on the patient's symptoms, age, etc. Force S, eye drops are usually administered 1 to 5 times a day, 1 to 5 drops per time. good. In the case of ophthalmic ointments, an appropriate amount is usually applied to each conjunctival sac 1-3 times a day for use. The disease targeted by the therapeutic agent for corneal wounds of the present invention is not particularly limited as long as it causes an abnormality in the PG composition of the corneal stroma. For example, corneal disorders associated with endogenous diseases such as keratoconus keratopathy, surgery, etc. Later, there may be mentioned drug-induced, trauma, exogenous diseases caused by contact lens wearing, and the like.

The above keratokeratosis is a disease in which the central part of the cornea becomes thinner and weaker, but it has been found that general keratokeratosis is based on abnormalities of GAG of PG in the corneal stroma. It has been shown that keratoconus has abnormally increased PG levels compared to normal corneas, and that the abnormalities are due to a decrease in keratan sulfate and an abnormal increase in chondroitin Z dermatan sulfate. It has been concluded to be a secondary phenomenon (Yue, BYJT, et al .: Histoch emica 1 studies of keratoconus. Curr. Eyeres., 7, 81-86, 1988., Sawa guchi, S., eta 1 .: Proteoglycan mo leculesinkeratoconusc ornea s. Inves t. Ophthal. Vis. Sci., 32, 1846—1853, 1991, Fun derbugh, J. L., eta 1 .: Alteredkeratansulfatee pitopesinkeratoconus., Invest. Ophthal.Vis. Sci., 30, 2278-2281, 1989). The therapeutic agent for corneal wounds of the present invention can normally disturb the actual PG composition disorder caused by injuring a wound on any part of the cornea, but when the corneal stroma is directly injured It can also be used as a therapeutic agent for corneal stromal wounds. In addition, the therapeutic agent for corneal wounds of the present invention can prevent the disorder of the composition of substantial PG and can be used as a preventive agent. BEST MODE FOR CARRYING OUT THE INVENTION

 Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to only these Examples.

(Example 1)

Therapeutic effect of perinastatin on physical injury of rat cornea

 After receiving SD male rats (7 weeks old, manufactured by Charles River Japan Co., Ltd.), they were preliminarily reared for 3 weeks. After that, Nemptal Injection (Dynabot) was intraperitoneally administered (40 mg / kg) for general anesthesia, and a drop of Ophthalmic Surface Anesthetic ぺ Nokiseal 0.4% solution (Santen Pharmaceutical) was also added. After instillation, all layers of the corneal epithelium from the limbus of the eye to the center were physically peeled using a microsurgical blade. Immediately after detachment of all layers, perinastatin (lot 67B8; 1500 uni.t / mL) was instilled 6 times a day at an interval of 5 μL per eye for about 2 hours. In addition, physiological saline was similarly instilled as a control group.ゥ Linastatin was instilled in one eye for each group in the control group, and one eye was used.

All layers peeled 0, 12, 24, 36, 48 At each hour after 8 hours, i.e., after all layers peeled, no eye drops, 6 times eye drops, 6 times eye drops 12 hours left, 12 times eye drops An intraperitoneal injection of an excessive amount of Nembutal injection solution was given to 4 mice in each group, which was left for 12 hours after instillation, and after lethality, the eyeball was excised and the fixative solution (1% glutaraldehyde '2.5 The solution was placed in% formalin / 0.15 M phosphate buffer, pH 7.2), shaken for 4 hours while changing the fixative three times, and an incision was made from the cornea to the vicinity of the retina. Further, the cells were fixed for 3 days while exchanging the fixative several times, then formalin was removed with PBS, and the cells were stored in 70% ethanol. Corneal tissue sections were prepared from the preserved cornea, and various GAG analyzes were performed according to the method of Koshiishi et al. (Ko shiishi I., Etal.: S tructural characterist icofoversulfatedchond roitin / de rma tansulfatesinthefibro uslesionsoftheliverwi thcirrhosis. m. Biophys., 370, 151-155, 1999).

 That is, a continuous section embedded in polyester wax was fixed and adjusted, and after degassing with ethanol, the sections were sequentially washed with 99%, 95%, 70% ethanol and distilled water.

 o 00 o

 CO

Replaced with 1M Tris—Aceta t e Bufffer (pH 8.0). After replacement, Chondroitinase ABC (0.5 uZmL), Chondroitinase AC II (0-Su / mL) ^ Co 11 agenase (0.5 μg / mL), Δ di -UA2 S (1 μg GAG released from digestion of the tissue with, for example, / mL) was analyzed as an unsaturated disaccharide by a fluorescent post-column high-performance liquid chromatography method using 2-cyanoacetamide as a fluorescent detection reagent. The results are shown in Tables 1, 2 and 3 for hyaluronic acid, chondroitin tetrasulfate and keratan sulfate.

Hyaluronic acid content (ngZrnm ² )

 Time course Treatment Pelinastatin administration group Saline administration group Untreated normal corneal stroma

0 No eye drops 6.41

 12 6 instillations 17.16

 24 6 times instillation 12:00 leave 7.42 10.40 7.54

 36 12 instillations 10.25

48 12 instillations 12:00 Leave 9.63 Table 2

表 3

Table 3

上記のごとく、 ラット角膜創傷後のいずれの GAG分子種においても、 ゥリナ スタチン点眼群は、 正常角膜の GAG含量の値に戻している力 \ 又は、 正常化を 促進している とが明らかとなった。 特に、 ゥリナスタチンは正常角膜に重要な ケラタン硫酸量の変動を是正する効果を有することより、 角膜の創傷の治療に有 効であると考えられる。 ケラタン硫酸の量が一時的に増加しているが、 S a i k a Sらが、 角膜上皮治癒の初期段階に いてルミカンは偏在的、 一時的に発現 し、 角膜上皮の接着 '移動による治癒促進に働いていることを報告している （S a i k a . , e t a l . : R o i e o f l um i c a n i n t h e c o r n e a l e p i t h e l i um d u r i n g wo u n d h e a l i n g. J - B i o l . C h e m. , 2 7 5, J a n, 2 6 0 7 - 2 6 1 2, 2 00 0) ことから、 表 3に見られるケラタン硫酸量の一時的な増加はケラタン硫 酸を構成糖とするルミカンが一時的に発現したことの影響があったものと考えら れる。 (実施例 2)

As described above, in any GAG molecular species after rat corneal injury, it is clear that the perinastatin ophthalmic group promotes normalization of normal cornea to GAG content and promotes normalization. Was. In particular, perinastatin is considered to be effective in treating corneal wounds because it has the effect of correcting fluctuations in the amount of keratan sulfate, which is important for normal corneas. Although the amount of keratan sulfate is increasing temporarily, Saika S et al. Reported that lumican is ubiquitously and temporarily expressed during the early stage of corneal epithelial healing, and promotes healing by corneal epithelial adhesion and migration. (S aika., Etal.: Roieofl um icaninthecornealepith eli um during wo undhealin g. J-B iol. Chem., 2 75, J an, 2 6 0 7-2 (6 1, 2, 2000) Therefore, it is considered that the temporary increase in the amount of keratan sulfate shown in Table 3 was due to the effect of the temporary expression of lumican having keratan sulfate as a constituent sugar. It is. (Example 2)

Therapeutic effect of perinastatin on physical injury (corneal epithelial detachment injury) of diabetic rat cornea

 1) Daily changes in corneal keratan sulfate content

Male SD rats (6-week-old, manufactured by Nippon Charlriva Co., Ltd.) were received and reared for 2 weeks. Thereafter, after fasting, 6 Omg / kg of streptozotocin (manufactured by Sigma) dissolved in 3 mM citric acid (pH 4.5) was administered from the tail vein of lmLZkg. Control (normal) rats were similarly administered the same buffer alone. The animals were bred normally for 2 weeks after administration, blood was collected from the tail vein, and the glucose level in the serum was measured using Glucose Test Co., Ltd. (manufactured by Wako Pure Chemical Industries, Ltd.). Was sorted out. Thereafter, diabetic rats and control rats were intraperitoneally administered with Nembutal injection solution (manufactured by Dynapot) (40 mg Zkg) to give general anesthesia. After instilling one drop of corneal epithelium, all layers of the corneal epithelium from the limbus to the center were physically separated using a microsurgical blade. Immediately after detachment of all layers, perinastatin (lot 67B8; 1500 units / mL) was instilled at a rate of 5 / x L per eye at an interval of about 2 hours 6 times a day. Physiological saline was similarly instilled in the control eyes of the control rats and the eyes of the control rats. One eye was used as a group of 20 diabetic rats in each of the perinastatin instillation group, the control group of the rats, and the control group of normal rats.

At 0 hour, 2 days, 1 week, 2 weeks, and 3 weeks after full-thickness detachment, an excessive amount of Nembutal injection was intraperitoneally administered to 4 mice in each group, and the mice were sacrificed. Immediately put in fixative solution (1% glutaraldehyde2.5% formalin / 0.15M phosphate buffer, PH 7.2), shake for 4 hours while changing fixative solution three times, and apply the solution from the cornea to the retina. An incision was made. Further, the cells were fixed for 3 days while exchanging the fixing solution several times, and then formalin was removed with PBS and stored in 70% ethanol. Thereafter, keratan sulfate and dermatan sulfate were measured by the method described in Example 1. Table 4 shows the measurement results of the corneal keratan sulfate content. Table 4

上記のごとく、 上皮損傷後早期 （損傷 2日後） にケラタン硫酸の量が一時的に 增加したが、 その増加量は糖尿病群では正常群に比べて低い。 しかし糖尿病群の 中でもゥリナスタチン投与群は生理食塩水点眼群に比べて高く、 正常群の値に近 かった。 S a i k aらが、 角膜上皮治癒の初期段階において ミカンは偏在的、 —時的に発現し、 角膜上皮の接着 ·移動による治癒促進に働いていることを報告 して ヽる ( S a i k a S . , e t a 1. : R o 1 e o f JU u m i c a n i n t h e Co r n e a l Ep i t h e l i um d u r i n g Wo u n d He a l i n g. J - B i o l . Ch em. , 275, J a ., 2607 -2612, 2000)。 更に、 F r i e n dらゃ F u k u s h i らは糖尿病ラ ットにおいて角膜上皮剥離後の創傷治癒速度の遅延が認められたと報告している (Fuk u s h i S . , e t a l . : Re e p i t h e l l a l i z a t i o n o f De nud e d Co r e a s i n D i a b e t i c Ra t s . Ex . Ey e R e s.， 31, 611— 621, 1 980, F r i e ri d J e t a l . : C o r n e a l Ep i t h e l i a l Ch a n g e s i n D i a b e t i c R a t s . Op h t h a l m i c R e s., 14， 269- 278, 1982 )。 これらより、 ケラタン硫酸量の一時的な增 加はケラタン硫酸を構成糖とするルミカンが一時的に発現したことの影響で、 糖 尿病ラットは正常ラットに比べてその発現量が低いために創傷治癒速度が遅延し . たものと考えられる。 更に、 ゥリナスタチンはルミカンの発現量を増やし正常ラ ットに近づけることで創傷治癒を促進すると考えられる。 2) 角膜デルマタン硫酸含量の経日変化

As described above, the amount of keratan sulfate temporarily increased early after epithelial injury (two days after injury), but the increase was lower in the diabetic group than in the normal group. However, among the diabetic groups, the pelinastatin-administered group was higher than the saline instilled group, and was close to the normal group. Saika et al. Report that oranges are ubiquitously and temporally expressed in the early stage of corneal epithelial healing and work to promote healing by adhesion and migration of corneal epithelium (Saika S., eta 1 .: Ro 1 eof JU umicaninthe Co rneal Ep itheli um during W und Healing g. J-Biol. Chem., 275, Ja., 2607-2612, 2000). Furthermore, Friend et al. Fukushi et al. Reported that wound healing rate was delayed after corneal epithelial detachment in diabetic rats (Fukushi S., etal .: Re epithellalization of Denuded Coreasein D). Iabetic Rats. Ex. Eye Res., 31, 611—621, 1980, Fri ri d J etal .: Corneal Ep ithelial Channels D iabetic Rats. Ophthalmic Res., 14, 269-278, 1982). From these results, the temporary increase in the amount of keratan sulfate was caused by the temporary expression of lumican, which is a constituent sugar of keratan sulfate. It is probable that the healing rate was delayed. In addition, perinastatin is thought to promote wound healing by increasing the expression level of lumican and approaching normal rats. 2) Daily changes in corneal dermatan sulfate content

Whole cornea, corneal quality and endothelium, the results of measurements dermatan sulfate content of _t corneal epithelium is shown in Table 5, 6, 7. Table 5

After day changes in corneal dermatan sulfate content after diabetic rats corneal abrasion (n gZmm ²⁾

表 6

Table 6

 Daily changes in corneal stroma and endothelial dermatan sulfate content (ng / mm2) after detachment of corneal epithelium in diabetic rats

 After epithelial detachment Diabetic rats Days of normal rat ゥ Rinastatin point Saline instillation group Saline instillation group

 Eye group

 Before peeling 29.2 ± 4.1 29.2- ± 4.1 19.4 ± 8.3

 Immediately after peeling 37.6 ± 3.4 37.6 ± 3.4 33.3 ± 5.5

 2 43.7 ± 4.6 42.2 ± 1.7 49.2 Sat 0.9

 7 28.0 ± 1.9 42.9 ± 1.4 22.6 ± 9.1

 14 29.6 ± 2.2 42.5 Sat 7.0 20.2 ± 2.3

21 25.0 ± 5.2 46.0 ± 4.6 24.6 ± 3.0 Table 7

Daily changes in corneal epithelial dermatan sulfate content (ng / mni ² ) after corneal epithelial detachment in diuretic rats

表 5、 6及ぴ 7から明らかなように角膜全体、 角膜実質 -内皮いずれにおいて も糖尿病群の生理食塩水点眼群でデルマタン硫酸が創傷後、 経日的に増加してい るのが認められた。 し力 し、 糖尿病群の U T I点眼群は、 正常ラット生理食塩水 点眼群と同様に、 創傷後、 一時的に急増するが、 一週間後には正常値に戻り、 以 後、 両群ともに正常値を 3週間持続することが認められた。 即ち、 糖尿病群に U T Iを点眼することにより、 角膜組織の破壌により一時的に增加したデコリンが 経曰的に正常群のそのデコリン量に近づいて行くと考えられる。 一方、 角膜上皮 の場合は、 修復過程において必要な量のデコリンの産生が糖尿病においては遅れ るが、 U T Iを点眼することにより正常な生合成が促進されると考えられる。 こ れらのことから、 糖尿病ラッ卜角膜損傷ではその治癒過程が異常をきたしている にも拘らず、 U T I点眼によって修復過程が正常に進むことを示唆し、 U T Iは 角膜創傷治療剤として効果が期待できる。 産業上の利用可能性

As is clear from Tables 5, 6 and 7, dermatan sulfate was observed to increase daily after wounding in the saline group in the diabetic group and in the corneal stroma and endothelium in all of the cornea. . In the diabetic group, the UTI ophthalmic group, like the normal rat saline ophthalmic group, temporarily increased after wounding, but returned to normal one week later, and then normal in both groups. For 3 weeks. That is, by instilling UTI in the diabetic group, it is considered that decorin temporarily added due to corneal tissue rupture approaches the amount of decorin in the normal group. On the other hand, in the case of corneal epithelium, the production of decorin required in the repair process is delayed in diabetes, but it is thought that normal biosynthesis is promoted by instilling UTI. These results suggest that, despite the abnormal healing process in diabetic rat corneal injury, the repair process proceeds normally with UTI instillation, and that UTI is effective as a therapeutic agent for corneal wounds. Can be expected. Industrial applicability

 According to the present invention, the physical and chemical corneal damage, including the subepithelial basement membrane, Bowman's membrane, corneal stroma, collagen-fibrous matrix, and Descemet's membrane, which exists between the corneal epithelium and the corneal endothelium In addition, a highly effective preventive / therapeutic agent having an excellent effect on mechanical damage and intrinsic corneal injury can be obtained.

Claims

The scope of the claims 1. A therapeutic agent for a corneal wound having an effect of returning a changed composition of a proteoglycan of a corneal stroma to a normal state, characterized by comprising perinastatin as an active ingredient. 2. A therapeutic agent for corneal stromal wounds, comprising perinastatin as an active ingredient. 3. The therapeutic agent for a corneal stromal wound according to claim 2, which has an effect of returning the composition of the changed proteodalican of the corneal stromal to a normal state.