JP3603129B2 - Therapeutic agent for diabetic keratosis - Google Patents

Description

【００１８】
以上のように糖尿病の最も簡便な動物モデルであるガラクトース血症ラット
の角膜内皮細胞障害修復遅延に対し化合物Ａ投与は改善せしめ、臨床的にも糖
尿病性角膜症を治療することが強く示唆された。
【００１９】
製剤例等
化合物Ａ；（２Ｓ，４Ｓ）−６−フルオロ−２’，５’− ジオキソスピロ［クロマン−４，４’−イミダゾリン］−２− カルボキサミド
【００２０】
製剤例１
処方Ａ
１００ｍｌ中
化合物Ａ ０．１０ｇ
リン酸一水素ナトリウム ０．７６ｇ
リン酸二水素ナトリウム ０．１６ｇ
塩化ナトリウム ０．４２ｇ
塩化ベンザルコニウム ０．０１ｇ
滅菌精製水 適 量
【００２１】
製法
滅菌精製水８０ｍｌにリン酸二水素ナトリウム、リン酸一水素ナトリウム、塩化ベンザルコニウムを加えて溶解した後、化合物Ａを加える。
化合物Ａを溶解させた後、滅菌精製水を加えて全量を１００ｍｌ とし、点眼剤に供する。
【００２２】
処方Ｂ
１００ｍｌ中
化合物Ａ ０．１０ｇ
リン酸一水素ナトリウム ０．７６ｇ
リン酸二水素ナトリウム ０．１６ｇ
塩化ナトリウム ０．４０ｇ
塩化ベンザルコニウム ０．０１ｇ
ショ糖 ０．０５ｇ
滅菌精製水 適 量
【００２３】
製法
処方Ａと同様の方法で、点眼剤を製した。
【００２４】
製剤例２
下記の表２に示される電荷調整剤０．４ｇと、精製卵黄レシチン１．６ ｇ と、下記の表３に示される油性成分８．０ｇとをクロロホルム／メタノール（５／１，Ｖ／Ｖ ）混液５０ｍｌ中で混合溶解した後、化合物Ａを０．１ 〜１．０ｇを加え、溶媒を完全に除去する。これに滅菌精製水９０ｇ を添加し、マイクロフルイタイザーにより高圧（１５００Ｋｇ／ｃｍ ^２ ）乳化し、１０％（Ｗ／Ｗ）脂肪乳剤とする。
【００２５】
処方
１００ｍｌ中
脂肪乳剤 ５０．０ｇ
リン酸一水素ナトリウム ０．７６ｇ
リン酸二水素ナトリウム ０．１６ｇ
塩化ナトリウム ０．４０ｇ
塩化ベンザルコニウム ０．０１ｇ
ショ糖 ０．０５ｇ
滅菌精製水 適 量
【００２６】
製法
処方Ａと同様の方法で、点眼剤を製した。
【００２７】
【表２】

【００２８】
【表３】

【００２９】

【００３０】
製法
化合物Ａとショ糖をエタノール／水（１／１，Ｗ／Ｗ）で練合し、乾燥後、結晶セルロース、乳糖を添加し、通常製剤に供される滑沢剤を加え、常法により錠剤、カプセル剤、顆粒剤等の経口製剤とする。
【００３１】

【００３２】
製法
製剤例３と同様の方法で経口製剤を製した。
【００３３】
【発明の効果】
糖尿病角膜症はその発生機序は未だ明らかではなく有効な治療薬も見出だされていないが、角膜上皮、知覚（シュワン細胞）および内皮細胞に種々の異常が認められる疾患であり、角膜の崩壊が重篤な場合，水泡性角膜症が発生し視機能を著しく障害する等しばしば難治性である。本発明は（２Ｓ，４Ｓ）−６−フルオロ−２’，５’− ジオキソスピロ［クロマン−４，４’−イミダゾリン］−２− カルボキサミドを有効成分とする糖尿病性角膜症治療剤であり、殊に経口、点眼剤形態を有し、糖尿病性角膜上皮症、糖尿病性角膜知覚低下、糖尿病性角膜内皮症に効果を有する。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a therapeutic agent for diabetic keratopathy comprising (2S, 4S) -6-fluoro-2 ', 5'-dioxospiro [chroman-4,4'-imidazoline] -2-carboxamide as an active ingredient, and in particular, The present invention relates to a composition which has an ophthalmic solution form and is used for diabetic corneal epitheliosis, diabetic corneal hyposensitivity, and diabetic corneal endotheliosis.
[0002]
[Prior art]
(2S, 4S) -6-Fluoro-2 ', 5'-dioxospiro [chroman-4,4'-imidazoline] -2-carboxamide is a compound discovered by the present applicant and has an effect on diabetic neuropathy. Because of its anti-ulcer action and drug for circulatory organs, it is currently in clinical trials as an oral drug because of its high long-term use safety. [EP-B-264586, U.S. Pat. S. 4,985,573 (Japanese Patent Publication No. 3-72227); S. 5,155,125 (JP-A-3-215435); S. 5,164,391 (Japanese Patent Laid-Open No. 3-106885), etc.
[Problems to be solved by the invention]
Diabetic keratopathy is a disease in which various abnormalities are observed in corneal epithelium, sensory (Schwann cells) and endothelial cells. When the corneal collapse is severe, it is often intractable, such as bullous keratopathy, which significantly impairs the visual function. However, the mechanism of its occurrence is not yet clear, and no effective therapeutic agent has been found.
[0004]
By the way, corneal endothelial cells have important functions but do not have regenerative ability or are very poor. However, diabetic patients develop simple diabetic retinopathy about 10 years after onset, and may eventually lead to proliferative retinopathy, retinal detachment, and blindness. It is said that the corneal endothelial cells are easily affected by strong stress such as application of photocoagulation, vitreous surgery or the like, or intraocular lens replacement for cataracts, to prevent keratosis.
[0005]
Regarding the fragility in the cornea of diabetic patients, when the endothelial cells of diabetic patients are observed by speculative microscopy even if they do not lead to keratosis, morphological abnormalities such as large and small, deformed, etc. were observed, suggesting the presence of diabetic corneal endothelial disorder. ing.
[0006]
The number of patients with diabetic keratopathy is not clearly reported, but Kitasato Ophthalmology Shimizu et al. Have an overwhelmingly high corneal damage rate of 56% in retinopathy patients and 17% in normal subjects, and corneal tissue is more vulnerable than normal subjects Reported that The number of diabetics is now potentially 6 million, and is expected to increase further in the future. The emergence of effective drugs will cause serious problems such as an increase in medical costs and impaired QOL for this disease. It is strongly desired.
[0007]
Means and method for solving the problem
SUMMARY OF THE INVENTION The present invention provides a method for treating diabetic keratopathy in which (2S, 4S) -6-fluoro-2 ', 5'-dioxospiro [chroman-4,4'-imidazoline] -2-carboxamide has no effective treatment. And provide a drug that is very useful in terms of patient medical expenses, welfare, and QOL (QOLITY OF LIFE).
[0008]
The pharmaceutical form of the therapeutic agent for diabetic keratosis of the present invention can be prepared using conventional pharmaceutical techniques, for example, as an oral preparation such as tablets, capsules, powders and granules, or as eye drops, injections, suppositories and the like. It can be used as a parenteral formulation. The dose varies depending on the symptoms, age, administration method, dosage form, etc. In the case of eye drops, it is usually 0.01% to 1% by instilling the eye drop once or several times, orally. In this case, the purpose can be achieved by administering to an adult in the range of 0.125 to 100 mg, preferably 0.5 to 2 mg per day in one or several divided doses. Hereinafter, the present invention will be described in more detail by way of examples.
[0009]
【Example】
Pharmacological pharmacology test example 1
Test method: The experiment was performed according to the method of Akagi et al. (Japanese Ophthalmology Bulletin 37, 809, 1986).
Using Sprague Dawley rats (15 rats per group) weighing 50 g (3 weeks old), the rats were divided into the following three groups. (1) The control group was a normal diet group for research (normal control drug), (2) a diet group containing 50% galactose (galactose alone group), and (3) a diet containing 50% galactose (2S, 4S)-. A group to which 6 mg / kg of 6-fluoro-2 ', 5'-dioxospiro [chroman-4,4'-imidazoline] -2-carboxamide (hereinafter abbreviated as compound A) was orally administered (compound A galactose group). The rats in each group were bred for about 6 weeks. For the freeze destruction of corneal endothelial cells, a 1.5 mm diameter stainless steel rod cooled to −70 ° C. in advance with acetone / dry ice was applied to the central part of the cornea of each group for 20 seconds to freeze-destruct endothelial cells transcorneally. After freezing and destruction, the cornea was excised with Nembutal lethal anesthesia over time and observed by the following method.
[0010]
Light microscopic observation: The cornea was fixed with 0.1 M phosphate buffer (pH 7.4) containing 1% paraformaldehyde and 1% glutaraldehyde and re-fixed with 1% osmium tetroxide, dehydrated with alcohol, and subjected to a predetermined method. Embedded in Epon resin. Light microscope sections were observed after toluidine blue-violet staining.
[0011]
Extension specimen observation: The cornea was left unfixed and immersed in a 60 mM 2Na EDTA buffer, and after fixation with methanol, only the endothelial cell layer was detached from the Descemet's membrane and attached to a slide glass. The sample was stained with toluidine blue and observed with a light microscope.
[0012]
result:
5 days after freeze disruption: In the normal control group, two to three layers of hill-like stratified regions were observed at the frozen site, and the endothelial cells were lightly restored. On the other hand, in the galactose-only group, several large circular bulges were found corresponding to the frozen area. The degenerated cells were present inside the bulge. In the compound A galactose group, a swelling structure was occasionally seen, but the size was small. In addition, in the stretched specimen, a number of nuclei were densely located in the central part of the cornea.
[0013]
7 days after freezing and destruction: In the normal control group and the compound A galactose group, no stratified site was observed at this time, and they were completely flattened and normalized. Even in this period, more than half of the galactose-only groups had bulging structures remaining. Compared to the image seen on day 5, the height of the overlay was reduced and the endothelial cells were composed of elongated, rather than circular, cytoplasm. In the stretched specimen image, dense and overlying nuclei were observed over a wide area.
[0014]
Pharmacological test example 2
Based on the results of pharmacological test example 1, clear utility of the present invention was observed.
The method of the experiment was carried out according to Example 1, and the corneas 5 days after the freeze destruction of (1) normal control group, (2) galactose alone group, and (3) compound A galactose group were extracted. The cornea of the rat's left eye was embedded in paraffin by a predetermined method, sliced, stained with hematoxylin and eosin, and observed under a light microscope.
[0015]
result:
The stratified site and raised structure of the corneal endothelial cells at the frozen site observed by light microscopy were considered to be in the repair process of corneal endothelial cell damage, and were observed in 25% of the normal control group 5 days after the freeze destruction. On the other hand, in the galactose-only group, 100% of all eyes were observed, the difference was statistically significant (P <0.05), and the repair of corneal endothelial cell damage was delayed. On the other hand, in the compound A galactose group, the stratified site and the raised structure of the corneal endothelial cells were observed in 22% of the eyes, and were statistically significantly (P <0.01) significantly normalized compared to the galactose alone group. Was. It has also been statistically shown that administration of Compound A improves the delayed repair of corneal endothelial cell damage in this model.
[0016]
Results of Test Example 2 (Table 1)
Effect on delayed repair of frozen-ruptured corneal endothelial cells in galactosemia rats [observation results 5 days after freeze-rupture]
[0017]
[Table 1]

[0018]
As described above, Compound A administration improved the delayed repair of corneal endothelial cell damage in galactosemia rats, the simplest animal model of diabetes, and strongly suggested that diabetic keratosis could be treated clinically. .
[0019]
Formulation Example Compound A; (2S, 4S) -6-fluoro-2 ′, 5′-dioxospiro [chroman-4,4′-imidazoline] -2-carboxamide
Formulation Example 1
Formula A
0.10 g of compound A in 100 ml
Sodium monohydrogen phosphate 0.76g
Sodium dihydrogen phosphate 0.16g
0.42 g of sodium chloride
Benzalkonium chloride 0.01g
Appropriate amount of sterilized purified water [0021]
Production Method Sodium dihydrogen phosphate, sodium monohydrogen phosphate and benzalkonium chloride are added to and dissolved in 80 ml of sterilized purified water, and then compound A is added.
After dissolving Compound A, sterile purified water is added to make a total volume of 100 ml, which is then used as eye drops.
[0022]
Formula B
0.10 g of compound A in 100 ml
Sodium monohydrogen phosphate 0.76g
Sodium dihydrogen phosphate 0.16g
0.40 g of sodium chloride
Benzalkonium chloride 0.01g
Sucrose 0.05g
Appropriate amount of sterilized purified water [0023]
Production Method An eye drop was prepared in the same manner as in Formulation A.
[0024]
Formulation Example 2
0.4 g of the charge control agent shown in Table 2 below, 1.6 g of purified egg yolk lecithin, and 8.0 g of the oily component shown in Table 3 below were mixed with chloroform / methanol (5/1, V / V). After mixing and dissolving in 50 ml of the mixed solution, 0.1 to 1.0 g of compound A is added, and the solvent is completely removed. 90 g of sterilized purified water is added thereto, and emulsified under high pressure (1500 Kg / cm ² ) with a microfluidizer to obtain a 10% (W / W) fat emulsion.
[0025]
Formulation 100ml fat emulsion 50.0g
Sodium monohydrogen phosphate 0.76g
Sodium dihydrogen phosphate 0.16g
0.40 g of sodium chloride
Benzalkonium chloride 0.01g
Sucrose 0.05g
Appropriate amount of sterilized purified water [0026]
Production Method An eye drop was prepared in the same manner as in Formulation A.
[0027]
[Table 2]

[0028]
[Table 3]

[0029]

[0030]
Formulation Compound A and sucrose are kneaded with ethanol / water (1/1, W / W), dried, and crystalline cellulose and lactose are added. Oral preparations such as tablets, capsules and granules.
[0031]

[0032]
An oral preparation was produced in the same manner as in Preparation Example 3.
[0033]
【The invention's effect】
The mechanism of diabetic keratopathy has not yet been elucidated, and no effective therapeutic agent has been found. However, various abnormalities are observed in the corneal epithelium, sensory (Schwann cells) and endothelial cells. If the disintegration is severe, bullous keratopathy occurs and often impairs the visual function, and is often intractable. The present invention relates to a therapeutic agent for diabetic keratosis comprising (2S, 4S) -6-fluoro-2 ', 5'-dioxospiro [chroman-4,4'-imidazoline] -2-carboxamide as an active ingredient, and in particular, It has oral and ophthalmic forms and is effective for diabetic corneal epitheliosis, diabetic corneal hypoperception, and diabetic corneal endotheliosis.

Claims (2)

A therapeutic agent for diabetic keratosis comprising (2S, 4S) -6-fluoro-2 ', 5'-dioxospiro [chroman-4,4'-imidazoline] -2-carboxamide as an active ingredient. The therapeutic agent for diabetic keratopathy according to claim 1 , which is in the form of an oral preparation or eye drops, and is used for diabetic corneal epitheliosis, diabetic corneal hypoesthesia or diabetic corneal endotheliosis.