JP2004182701A - Ocular tissue vascularization inhibitor containing hydroxamic acid derivative as active ingredient - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new pharmacological activity regarding a hydroxamic acid derivative. <P>SOLUTION: This hydroxamic acid derivative is examined for a vascularization inhibitory activity by using a krypton laser-induced rat choroidea vascularization method, and found out that the hydroxamic acid derivative having hydrophilic group has an excellent vascularization inhibitory activity. Therefore, it is useful as a preventing and treating agent of ocular diseases associated with the vascularization, especially, diabetic retinopathy, premature baby retinopathy, macular degeneration, neovascular glaucoma, occlusion of retinal vein, occlusion of retinal artery, pterygium, rubeosis, vascularization in cornea, etc. <P>COPYRIGHT: (C)2004,JPO&NCIPI

Description

【００１３】
［式中、Ｒ^１は、水素、水酸基、アリール（Ｃ_１−Ｃ_６）アルキレン基または−Ａ−ＳＯｎ−Ｂ基（Ａは、（Ｃ_１−Ｃ_６）アルキレン基を表し、Ｂは、（Ｃ_１−Ｃ_６）アルキル基、（Ｃ_１−Ｃ_６）アシル基、アリール基または硫黄、酸素又は窒素のヘテロ原子を少なくとも１個有する５〜６員環骨格を表し、ｎは０，１または２のいずれかの数を表す。）を表し、Ｒ^２は、水素、（Ｃ_１−Ｃ_６）アルキル基、（Ｃ_１−Ｃ_６）アルキルオキシ基または（Ｃ_１−Ｃ_６）アルキルチオ基を表し、Ｒ^３及びＲ^４は、同一又は異なっていてよく、水素、（Ｃ_１−Ｃ_６）アルキル基、（Ｃ_３−Ｃ_１０）シクロアルキル基、アリール基またはアリール（Ｃ_１−Ｃ_６）アルキレン基を表し、Ｒ^５は親水性基を表し、Ｒ^６は、水素、未置換のベンジル基、トリアルキルシリル基、ｔｅｒｔ−ブチルジフェニルシリル基、テトラヒドロピラニル基またはｔｅｒｔ−ブチル基を表す。］
以下、本発明を詳細に説明する。
【００１４】
一般式（Ｉ）中のＲ^５が親水性基であるので、本化合物は、水溶性が向上し、高濃度の薬剤調製も可能であり、注射剤や点眼剤などの水性製剤として適用できる。
【００１５】
親水性基Ｒ^５としては、−Ｙ−Ｃ基またはＣ基（Ｙは（Ｃ_１−Ｃ_６）アルキレン基、酸素、イミノ基、（Ｃ_１−Ｃ_６）アルキレンイミノ基を表し、Ｃはスルホン酸基、ホスホン酸基、アミジノ基、（Ｃ_１−Ｃ_６）アシル基、アシルイミドイル基、ジホスホノメチン基、ジカルボキシメチン基またはテトラ（Ｃ_１−Ｃ_６）アルキルビスホスホノメチルイミノ基を表す。但し、Ｙが（Ｃ_１−Ｃ_６）アルキレン基であるとき、Ｃは（Ｃ_１−Ｃ_６）アシル基でない。）が挙げられる。より好ましいＲ^５の親水性基としては、硫酸基、スルホン酸基、リン酸基、グアニド基、グアニドメチレン基、アミジノ基、アセトイミドイルイミノメチレン基、プロピオンイミドイルイミノメチレン基、ベンズイミドイルイミノメチレン基、アセトアミドメチレン基またはテトラエチルビスホスホノメチルイミノ基が挙げられ、さらに好ましくは硫酸基、スルホン酸基、リン酸基、グアニド基、グアニドメチレン基またはテトラエチルビスホスホノメチルイミノ基である。
【００１６】
一般式（Ｉ）で表される化合物中の各基は、以下に示す通りである。
【００１７】
スルホン酸基とは−ＳＯ_３Ｈを示し、硫酸基とは−ＯＳＯ_３Ｈを示す。ホスホン酸基とは−ＰＯ_３Ｈ_２を示し、リン酸基とは−ＯＰＯ_３Ｈ_２を示す。アミジノ基とは−Ｃ（＝ＮＨ）ＮＨ_２を、グアニド基とは−ＮＨ−Ｃ（＝ＮＨ）ＮＨ_２を示す。アミノメチレン基とは−ＣＨ_２ＮＨ_２を、グアニドメチレン基とは−ＣＨ_２−ＮＨ−Ｃ（＝ＮＨ）ＮＨ_２を示す。アセトアミドメチレン基とはＣＨ_２ＮＨ−ＣＯＣＨ_３を、アセトイミドイルイミノメチレン基とは−ＣＨ_２−ＮＨ−Ｃ（＝ＮＨ）ＣＨ_３を、プロピオンイミドイルイミノメチレン基とは−ＣＨ_２−ＮＨ−Ｃ（＝ＮＨ）ＣＨ_２ＣＨ_３を、ベンズイミドイルイミノメチレン基とは−ＣＨ_２−ＮＨ−Ｃ（＝ＮＨ）Ｃ_６Ｈ_５を示す。ジホスホノメチン基とは−ＣＨ［ＰＯ（ＯＨ）_２］_２を、ジカルボキシメチン基とは−ＣＨ（ＣＯ_２Ｈ）_２を示す。
【００１８】
（Ｃ_１−Ｃ_６）アルキル基とは１−６の炭素原子を有する直鎖又は分岐鎖のアルキル基を示し、メチル基、エチル基、ｎ−プロピル基、ｉｓｏ−プロピル基、ｎ−ブチル基、ｓｅｃ−ブチル基、ｔｅｒｔ−ブチル基、ペンチル基、ヘキシル基等が含まれる。
【００１９】
（Ｃ_１−Ｃ_６）アルキレン基とは−（ＣＨ_２）_ｎ−（ｎ＝１−６）を示す。イミノ基とは−ＮＨ−を示す。
【００２０】
（Ｃ_１−Ｃ_６）アルキレンイミノ基とは−（ＣＨ_２）_ｎ−ＮＨ−（ｎ＝１−６）を示し、好ましくは（Ｃ_１−Ｃ_３）アルキレンイミノ基であり、より好ましくは−ＣＨ_２−ＮＨ−である。
【００２１】
（Ｃ_１−Ｃ_６）アシル基とは１−６の炭素原子を有する直鎖又は分岐鎖のアルキルアシル基を示し、ホルミル基、アセチル基、ｎ−プロパノイル基、ｎ−ブタノイル基等が含まれる。
【００２２】
（Ｃ_３−Ｃ_１０）シクロアルキル基は、単環式のものであっても、二環式などの複数の環の縮合した形態のものであってよく、その炭素数は３−１０、好ましくは３−７、より好ましくは３−６である。例えば、シクロプロピル基、シクロブチル基、シクロペンチル基、シクロヘキシル基、シクロヘプチル基、 ［２．２．１］ビシクロヘプチル基等が挙げられる。
【００２３】
アシルイミドイル基とは、−Ｃ（＝ＮＨ）−［（Ｃ_１−Ｃ_６）アルキル］または−Ｃ（＝ＮＨ）−（アリール）を示す。
【００２４】
硫黄、酸素、又は窒素のヘテロ原子を少なくとも一つ有する５〜６員環骨格としては、例えば好ましいものとしてチエニル基、チアゾリル基、イミダゾリル基、ピリジル基等が挙げられる。
【００２５】
アリール基とは、置換されてもよいフェニル基、ナフチル基、アントラセニル基等の芳香環を示す。
【００２６】
アリール基上の置換基としては、（Ｃ_１−Ｃ_６）アルキル基、（Ｃ_１−Ｃ_６）アシル基、水酸基、アミノ基、カルボキシル基、ハロゲン等が含まれ、また二つ以上の置換基を有する場合はこれらを組み合わせたものでもよい。
【００２７】
ハロゲンはフッ素、塩素、臭素、ヨウ素原子を示す。
【００２８】
また、一般式（Ｉ）の化合物の塩としてはナトリウム、カリウム等のアルカリ金属、マグネシウム、カルシウム等のアルカリ土類金属、塩化水素酸、臭化水素酸等の鉱酸あるいはアンモニアよりなる無機塩と、モルホリン、ピペリジン、ジメチルアミン、ジエチルアミン、酢酸、クエン酸、シュウ酸等の有機塩が含まれる。また溶媒和物としては、例えば水和物が挙げられる。
【００２９】
上記一般式（Ｉ）で表される化合物には三つの不斉炭素原子が存在するため、本発明は、一般式（Ｉ）のジアステレオマー、それらの混合物であるラセミ体を包含する。
【００３０】
本化合物のより好ましい具体例としては、下記の化合物またはそれらの立体異性体、製薬学的に許容される塩若しくは溶媒和物が挙げられる。
【００３１】
▲１▼ Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−（３−フェニルトリメチレン）サクシニル］−Ｌ−４’−グアニドフェニルアラニン−Ｎ−メチルアミド 塩酸塩
【化３】

【００３２】
▲２▼ Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−ヒドロキシサクシニル］−Ｌ−４’−グアニドフェニルアラニン−Ｎ−メチルアミド １酢酸塩
【化４】

【００３３】
▲３▼ Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−（３−フェニルトリメチレン）サクシニル］−Ｏ−スルホ−Ｌ−チロシン−Ｎ−１−ナフチルアミド １ナトリウム塩
【化５】

【００３４】
▲４▼ Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−ヒドロキシサクシニル］−４’−［テトラエチルビス（ホスホノ）メチルイミノ］−Ｌ−フェニルアラニン−Ｎ−シクロプロピルアミド
【化６】

【００３５】
上記一般式（Ｉ）で表されるヒドロキサム酸誘導体の有用性を調べるべく、眼組織における本化合物の血管新生阻害作用について検討した。詳細については後述の薬理試験の項で示すが、本化合物は、ｉｎ ｖｉｖｏ の眼組織血管新生評価モデルであるクリプトンレーザー誘発のラット脈絡膜血管新生法に対して強い阻害作用を示すことを見出した。この試験結果より、本化合物は、眼組織血管新生阻害剤および血管新生が関与する眼科疾患、特に、糖尿病性網膜症、未熟児網膜症、黄斑変性症、血管新生緑内障、網膜静脈閉塞症、網膜動脈閉塞症、翼状片、ルベオーシス、角膜新生血管症などの予防剤、治療剤および抑制剤として有用である。
【００３６】
本化合物の投与は経口でも非経口でも行うことができるが、非経口投与、特に局所投与が好ましい。局所投与剤型としては、注射剤、点眼剤等が挙げられ、それらの製剤は汎用されている技術を用いて調製することができる。これらの局所投与製剤の調製には、必要に応じて塩酸、クエン酸、リン酸、酢酸、水酸化ナトリウム、炭酸ナトリウム、炭酸水素ナトリウムなどのｐＨ調節剤、エデト酸ナトリウムなどの安定化剤、グリセリン、プロピレングリコール、ポリエチレングリコール、塩化ナトリウム、塩化カリウム、ソルビトール、マンニトールなどの等張化剤、クエン酸、リン酸水素ナトリウム、氷酢酸、トロメタモールなどの緩衝剤、ベンジルアルコールなどの無痛化剤、塩化ベンザルコニウム、塩化ベンゼトニウム、パラオキシ安息香酸エステル、安息香酸ナトリウム、クロロブタノールなどの保存剤等を適量添加することができる。また、局所投与注射としては、結膜下注射、網膜下注射、硝子体内注射、強膜内注射、前房内注射、テノン嚢注射等が挙げられる。
【００３７】
本化合物の投与量は症状、年齢、剤型等によって適宜選択できるが、点眼剤であれば０．０００１〜１０％の濃度、より好ましくは０．００１〜１％の濃度のものを日に１回または数回点眼すればよい。
【００３８】
以下に薬理試験の結果を示すが、これらは本発明をよりよく理解するためのものであり、本発明の範囲を限定するものではない。
【００３９】
【実施例】
［１］クリプトンレーザー誘発ラット脈絡膜血管新生法による眼組織血管新生阻害試験
ｉｎ ｖｉｖｏ における脈絡膜血管新生モデルとして、クリプトンレーザーを用いた強度網膜光凝固を有色ラットに施し、Ｂｒｕｃｈ膜を断裂することにより脈絡膜血管由来の血管新生が誘発される動物モデルがＡｒｃｈ Ｏｐｈｔｈａｌｍｏｌ，１０７，２６４−２６９ （１９８９）に報告されている。そこで、上記文献に記載された方法に従って、上記動物モデルを作製し、Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｒ ｏｒ Ｓ）−（３−フェニルトリメチレン）サクシニル］−Ｌ−４’−グアニドフェニルアラニン−Ｎ−メチルアミド 塩酸塩（被験化合物Ａ）、Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−ヒドロキシサクシニル］−Ｌ−４’−グアニドフェニルアラニン−Ｎ−メチルアミド １酢酸塩（被験化合物Ｂ）、Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（ＲＳ）−（３−フェニルトリメチレン）サクシニル］−Ｏ−スルホ−Ｌ−チロシン−Ｎ−１−ナフチルアミド １ナトリウム塩（被験化合物Ｃ）およびＮ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−ヒドロキシサクシニル］−４’−［テトラエチルビス（ホスホノ）メチルイミノ］−Ｌ−フェニルアラニン−Ｎ−シクロプロピルアミド（被験化合物Ｄ）の脈絡膜血管新生に対する阻害効果を検討した。
【００４０】
（被験化合物溶液の調製）
被験化合物Ａ、被験化合物Ｂおよび被験化合物Ｄを、それぞれ１０％のポリオキシエチレン硬化ヒマシ油６０（以下「ＨＣＯ６０」とする。）を含有するリン酸緩衝液に溶解して６ｍｇ／ｍｌの各被験化合物溶液を得た。また、被験化合物Ｃを、１０％のＨＣＯ６０を含有するリン酸緩衝液に懸濁させて６ｍｇ／ｍｌの懸濁溶液を得た。
【００４１】
（実験方法）
有色ラットに麻酔薬を筋肉内投与することにより全身麻酔を行った。トロピカミドを点眼して散瞳させた後、クリプトンレーザー光凝固装置を使用し、スポットサイズ１００μｍ、出力１００ｍＷ、凝固時間０．１秒の条件にて、焦点を網膜深層に合わせて両眼の眼底後局部へ散在性に８箇所の光凝固を施した。各被験化合物を、光凝固直後から１日１回、３００μｇ／５０μｌ／ｅｙｅで両眼の結膜下に投与した。なお、コントロールとして、上記と同様の操作を行い被験化合物の代わりに１０％のＨＣＯ６０を含有するリン酸緩衝液を投与した。
【００４２】
（眼組織血管新生評価法）
光凝固１４日目に１０％フルオレセイン０．１ｍｌを尾静脈から注入して、蛍光眼底造影を行った。新生血管は各凝固スポットでの蛍光漏出を指標とし、蛍光漏出が認められないものを陰性、明らかな蛍光漏出が認められるものを陽性と判断した。また、若干の蛍光漏出が認められるものは、それが２ポイント存在した時に陽性とした。下記の計算式に従い眼組織における新生血管発現率を算出した。
【００４３】
新生血管発現率（％）
＝（蛍光漏出陽性ポイント数／光凝固ポイント数）×１００
各被験化合物による血管新生阻害率は下記計算式に従い算出した。
【００４４】
血管新生阻害率（％）
＝（１−被験化合物の新生血管発現率／コントロールの新生血管発現率）×１００
上記に従い算出した各被験化合物の血管新生阻害率を表１に示す（なお、表中の数字は８眼の平均値を示す）。
【００４５】
【表１】

【００４６】
［２］製剤処方例
注射剤（１００ｍｌ）
処方：Ｎ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−ヒドロキシサクシニル］−４’−［テトラエチルビス（ホスホノ）メチルイミノ］−Ｌ−フェニルアラニン−Ｎ−シクロプロピルアミド ５００ｍｇ

【００４７】滅菌精製水に上記処方量のＮ−［４−（Ｎ−ヒドロキシアミノ）−２（Ｒ）−イソブチル−３（Ｓ）−ヒドロキシサクシニル］−４’−［テトラエチルビス（ホスホノ）メチルイミノ］−Ｌ−フェニルアラニン−Ｎ−シクロプロピルアミドおよび塩化ナトリウムを溶解して１００ｍｌの注射剤を得た。
【００４８】
【発明の効果】
上記の薬理試験の結果から明らかなように、本化合物は眼組織において優れた血管新生阻害作用を有するので、糖尿病性網膜症、未熟児網膜症、黄斑変性症、血管新生緑内障、網膜静脈閉塞症、網膜動脈閉塞症、翼状片、ルベオーシス、角膜新生血管症などの血管新生が関与する眼科疾患の予防剤および治療剤として有用である。また、本化合物は、親水性基を有し、水に対する溶解性が改善されているので、注射剤、点眼剤などの水性製剤に適する。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to an ocular tissue angiogenesis inhibitor containing a hydroxamic acid derivative having a hydrophilic group as an active ingredient, and provides a drug useful for treating ophthalmic diseases.
[0002]
[Prior art]
Vascular homeostasis is maintained by various functions of endothelial cells. Vascular endothelial cells 1) mediate the transport of necessary components such as nutrients in blood to tissues and prevent the passage of unnecessarily large amounts of components. 2) Smoothly circulate without coagulating blood. It has an effect of preventing blood bleeding when a blood vessel is disconnected, and an effect of regulating blood vessel tension at a constant level.
[0003]
Angiogenesis occurs at the stage of degradation of basement membrane by proteases produced by vascular endothelial cells, migration and proliferation of vascular endothelial cells, formation of vascular endothelial cell lumen, formation of basement membrane and surrounding pericytes. In ocular tissues, blood vessels concentrate in the retina, and when angiogenesis occurs, diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein occlusion, retinal artery occlusion, pterygium, rubeosis In addition, in the cornea, which is an avascular tissue, vascularization may occur due to damage, and corneal neovascular disease may occur.
[0004]
Patent Documents 1 and 2 disclose that hydroxamic acid derivatives having an amide bond have an inhibitory action on matrix metalloproteinases (MMPs) and TNF-α converting enzyme, and are useful compounds for diseases related to tissue degradation such as arthritis. It is disclosed. Patent Document 3 describes a matrix metalloproteinase inhibitor. The active ingredient of the inhibitor has a different chemical structure from the active ingredient of the drug of the present invention, and has a hydrophilic property which is a feature of the present invention. There is no disclosure regarding the introduction of a functional group.
[0005]
However, there is no report on the therapeutic effect of a disease in ocular tissues based on the angiogenesis inhibitory action of a hydroxamic acid derivative having an amide group.
[0006]
[Patent Document 1]
International Publication WO96 / 33968.
[0007]
[Patent Document 2]
International Publication WO99 / 31052.
[0008]
[Patent Document 3]
International Publication WO93 / 21942.
[0009]
[Problems to be solved by the invention]
It is a very interesting problem to find a new pharmacological action for such a hydroxamic acid derivative useful as a medicine. There is also a need for a therapeutic agent for ophthalmic diseases that has good solubility in water and is suitable for aqueous preparations.
[0010]
[Means for Solving the Problems]
The present inventors conducted an ocular tissue angiogenesis inhibition test using a krypton laser-induced rat choroidal neovascularization method in order to find a new pharmacological action of the hydroxamic acid derivative. As a result, the hydroxamic acid derivative having a hydrophilic group of the present invention exerts an excellent angiogenesis inhibitory action on ocular tissues, so that diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein It has been found that it is useful as a therapeutic agent for ophthalmic diseases such as obstruction, retinal artery occlusion, pterygium, rubeosis, and corneal neovascular disease. Further, the hydroxamic acid derivative of the present invention has a hydrophilic group and improved solubility in water, and thus is suitable for aqueous preparations such as injections and eye drops.
[0011]
BEST MODE FOR CARRYING OUT THE INVENTION
The present invention relates to an ophthalmic tissue comprising a compound represented by the following general formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof (hereinafter, referred to as “the present compound”) as an active ingredient. It is intended to provide an angiogenesis inhibitor and a therapeutic agent for an ophthalmic disease involving angiogenesis. In addition, the production method of this compound is described in detail in WO96 / 33968 and WO99 / 31052.
[0012]
Embedded image

[0013]
[In the formula, R ¹ represents hydrogen, a hydroxyl group, an aryl (C ₁ -C ₆ ) alkylene group or an —A-SOn-B group (A represents a (C ₁ -C ₆ ) alkylene group; A C ₁ -C ₆ ) alkyl group, a (C ₁ -C ₆ ) acyl group, an aryl group or a 5- to 6-membered ring skeleton having at least one sulfur, oxygen or nitrogen heteroatom, wherein n is 0, 1 or R ² represents hydrogen, a (C ₁ -C ₆ ) alkyl group, a (C ₁ -C ₆ ) alkyloxy group or a (C ₁ -C ₆ ) alkylthio group. And R ³ and R ⁴ may be the same or different and represent hydrogen, a (C ₁ -C ₆ ) alkyl group, a (C ₃ -C ₁₀ ) cycloalkyl group, an aryl group or an aryl (C ₁ -C ₆ ) Represents an alkylene group, R ⁵ represents a hydrophilic group, and R ⁶ represents water A benzyl group, a trialkylsilyl group, a tert-butyldiphenylsilyl group, a tetrahydropyranyl group or a tert-butyl group. ]
Hereinafter, the present invention will be described in detail.
[0014]
Since R ⁵ in the general formula (I) is a hydrophilic group, the compound has improved water solubility, can be used for preparing a drug at a high concentration, and can be applied as an aqueous preparation such as an injection or an eye drop.
[0015]
The hydrophilic group ^{R 5,} -Y-C group or a C group (Y represents _(C 1 -C ₆₎ alkylene group, an oxygen, an imino _group, a _(C 1 -C ₆₎ alkyleneimino group, C is a sulfonic Represents an acid group, a phosphonic acid group, an amidino group, a (C ₁ -C ₆ ) acyl group, an acylimidoyl group, a diphosphonomethine group, a dicarboxymethine group or a tetra (C ₁ -C ₆ ) alkylbisphosphonomethylimino group However, when Y is a (C ₁ -C ₆ ) alkylene group, C is not a (C ₁ -C ₆ ) acyl group. More preferred hydrophilic groups of R ⁵ include a sulfuric acid group, a sulfonic acid group, a phosphoric acid group, a guanide group, a guanidomethylene group, an amidino group, an acetimidoyl imino methylene group, a propionimidoyl imino methylene group, and a benzimidoyl imino methylene group. Groups, an acetamidomethylene group or a tetraethylbisphosphonomethylimino group, more preferably a sulfate group, a sulfonic acid group, a phosphoric acid group, a guanide group, a guanidomethylene group or a tetraethylbisphosphonomethylimino group.
[0016]
Each group in the compound represented by the general formula (I) is as shown below.
[0017]
The sulfonic acid group shows a -SO ₃ H, shows the -OSO ₃ H is a sulfate group. The phosphonic acid group shows a _-PO 3 _{H 2,} shows a -OPO ₃ H ₂ and phosphate groups. The The amidino group -C (= NH) NH _2, shows a -NH-C (= NH) NH 2 and guanido groups. The _-CH 2 NH ₂ The aminomethylene group, a _{-CH 2 -NH-C (= NH} ) NH 2 and Guanidomechiren group. An acetamidomethylene group is CH ₂ NH—COCH ₃ , an acetimidoyliminomethylene group is —CH ₂ —NH—C (＝ NH) CH _3, and a propionimidoyliminomethylene group is —CH ₂ —NH—. C (= NH) a _CH 2 CH _3, showing the _{-CH 2 -NH-C (= NH} ) C 6 H 5 is a benzimidamide garment amino methylene group. What Is Jihosuhonomechin group _{-CH [PO (OH) 2]} 2, showing the -CH _{(CO 2 H)} 2 and dicarbonitrile Kishime Chin group.
[0018]
(C ₁ -C ₆ ) alkyl group means a linear or branched alkyl group having _{1 to 6} carbon atoms, and is a methyl group, an ethyl group, an n-propyl group, an isopropyl group, or an n-butyl group. , Sec-butyl, tert-butyl, pentyl, hexyl and the like.
[0019]
The (C ₁ -C ₆ ) alkylene group refers to — (CH ₂ ) _n — (n = 1-6). The imino group indicates -NH-.
[0020]
The (C ₁ -C ₆ ) alkyleneimino group refers to — (CH ₂ ) _n —NH— (n = 1-6), and is preferably a (C ₁ -C ₃ ) alkyleneimino group, and more preferably —. CH ₂ —NH—.
[0021]
The (C ₁ -C ₆ ) acyl group refers to a linear or branched alkylacyl group having _{1 to 6} carbon atoms, and includes a formyl group, an acetyl group, an n-propanoyl group, an n-butanoyl group and the like. .
[0022]
The (C ₃ -C ₁₀ ) cycloalkyl group may be a monocyclic group or a condensed form of a plurality of rings such as a bicyclic group, and has _{3 to 10} carbon atoms, preferably Is 3-7, more preferably 3-6. For example, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a [2.2.1] bicycloheptyl group and the like can be mentioned.
[0023]
The acylimidoyl _{groups, -C (= NH) - [} (C 1 -C 6) alkyl or -C (= NH) - shows the (aryl).
[0024]
As the 5- or 6-membered ring skeleton having at least one sulfur, oxygen or nitrogen heteroatom, for example, preferred are a thienyl group, a thiazolyl group, an imidazolyl group and a pyridyl group.
[0025]
The aryl group indicates an aromatic ring such as a phenyl group, a naphthyl group, and an anthracenyl group which may be substituted.
[0026]
Examples of the substituent on the aryl group include a (C ₁ -C ₆ ) alkyl group, a (C ₁ -C ₆ ) acyl group, a hydroxyl group, an amino group, a carboxyl group, and a halogen. When these have, these may be combined.
[0027]
Halogen represents a fluorine, chlorine, bromine or iodine atom.
[0028]
Examples of the salt of the compound of the general formula (I) include an alkali metal such as sodium and potassium; an alkaline earth metal such as magnesium and calcium; a mineral acid such as hydrochloric acid and hydrobromic acid; , Morpholine, piperidine, dimethylamine, diethylamine, acetic acid, citric acid, oxalic acid and the like. Examples of solvates include hydrates.
[0029]
Since the compound represented by the above general formula (I) has three asymmetric carbon atoms, the present invention includes a diastereomer of the general formula (I) and a racemate which is a mixture thereof.
[0030]
More preferred specific examples of the present compound include the following compounds or their stereoisomers, pharmaceutically acceptable salts or solvates.
[0031]
{Circle around (1)} N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S)-(3-phenyltrimethylene) succinyl] -L-4′-guanidephenylalanine-N-methylamide hydrochloric acid Salt

[0032]
{Circle around (2)} N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S) -hydroxysuccinyl] -L-4′-guanidephenylalanine-N-methylamide monoacetate

[0033]
{Circle around (3)} N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S)-(3-phenyltrimethylene) succinyl] -O-sulfo-L-tyrosine-N-1-naphthyl Amide monosodium salt

[0034]
{Circle around (4)} N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S) -hydroxysuccinyl] -4 ′-[tetraethylbis (phosphono) methylimino] -L-phenylalanine-N-cyclo Propylamide [Formula 6]

[0035]
In order to examine the usefulness of the hydroxamic acid derivative represented by the above general formula (I), the angiogenesis inhibitory effect of the present compound on eye tissues was examined. Although the details will be described in the section of pharmacological test described below, the present compound was found to exhibit a strong inhibitory effect on a krypton laser-induced rat choroidal neovascularization method, which is an in vivo ocular tissue angiogenesis evaluation model. From the results of this test, it is found that this compound is an ocular tissue angiogenesis inhibitor and ophthalmic diseases involving angiogenesis, especially diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein occlusion, and retinal vein occlusion. It is useful as a preventive, therapeutic and inhibitor for arterial occlusion, pterygium, rubeosis, corneal neovascular disease and the like.
[0036]
The compound can be administered orally or parenterally, but parenteral administration, particularly topical administration, is preferred. Examples of the dosage form for topical administration include injections, eye drops and the like, and these preparations can be prepared by using widely used techniques. Preparations of these topical preparations include, if necessary, hydrochloric acid, citric acid, phosphoric acid, acetic acid, sodium hydroxide, sodium carbonate, sodium bicarbonate and other pH regulators, sodium edetate and other stabilizers, glycerin Isotoning agents such as propylene glycol, polyethylene glycol, sodium chloride, potassium chloride, sorbitol, and mannitol; buffers such as citric acid, sodium hydrogen phosphate, glacial acetic acid, and tromethamol; soothing agents such as benzyl alcohol; Preservatives such as ruconium, benzethonium chloride, paraoxybenzoate, sodium benzoate and chlorobutanol can be added in appropriate amounts. Examples of the locally administered injection include subconjunctival injection, subretinal injection, intravitreal injection, intrascleral injection, anterior chamber injection, Tenon's capsule injection, and the like.
[0037]
The dose of the present compound can be appropriately selected depending on the condition, age, dosage form and the like. In the case of eye drops, a concentration of 0.0001 to 10%, more preferably a concentration of 0.001 to 1% per day is preferred. It may be administered once or several times.
[0038]
The results of the pharmacological tests are shown below, but these are for better understanding of the present invention and do not limit the scope of the present invention.
[0039]
【Example】
[1] Inhibition test of ocular tissue angiogenesis by krypton laser-induced rat choroidal neovascularization As a model of choroidal neovascularization in vivo, a colored rat is subjected to intense retinal photocoagulation using a krypton laser to rupture the Bruch's membrane. An animal model in which vascular-derived angiogenesis is induced has been reported in Arch Ophthalmol, 107, 264-269 (1989). Therefore, the above animal model was prepared according to the method described in the above literature, and N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (R or S)-(3-phenyltrimethylene) was used. ) Succinyl] -L-4′-guanidephenylalanine-N-methylamide hydrochloride (test compound A), N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S) -hydroxysuccinyl ] -L-4'-guanidophenylalanine-N-methylamide monoacetate (test compound B), N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (RS)-(3- Phenyltrimethylene) succinyl] -O-sulfo-L-tyrosine-N-1-naphthylamide monosodium salt (test compound C) and N- [4- (N-hydroxyamino) -2 (R ) -Isobutyl-3 (S) -hydroxysuccinyl] -4 '-[tetraethylbis (phosphono) methylimino] -L-phenylalanine-N-cyclopropylamide (test compound D) was examined for its inhibitory effect on choroidal neovascularization.
[0040]
(Preparation of test compound solution)
Test compound A, test compound B and test compound D were dissolved in a phosphate buffer containing 10% polyoxyethylene hydrogenated castor oil 60 (hereinafter referred to as "HCO60"), and each test compound was dissolved at 6 mg / ml. A compound solution was obtained. Test compound C was suspended in a phosphate buffer containing 10% HCO60 to obtain a 6 mg / ml suspension.
[0041]
(experimental method)
General anesthesia was performed by intramuscularly administering an anesthetic to colored rats. After instilling tropicamide and causing mydriasis, using a krypton laser photocoagulator, focus on the deep retina under the conditions of a spot size of 100 μm, an output of 100 mW, and a coagulation time of 0.1 second. Eight spots of photocoagulation were scattered locally. Each test compound was administered once a day immediately after photocoagulation at a dose of 300 μg / 50 μl / eye under the conjunctiva of both eyes. As a control, a phosphate buffer containing 10% HCO60 was administered instead of the test compound by performing the same operation as described above.
[0042]
(Eye tissue angiogenesis evaluation method)
On day 14 of photocoagulation, 0.1 ml of 10% fluorescein was injected from the tail vein to perform fluorescent fundus imaging. For the new blood vessels, the fluorescence leak at each coagulation spot was used as an index, and those without fluorescence leakage were judged as negative, and those with obvious fluorescence leak were judged as positive. In addition, a sample in which some fluorescence leakage was observed was regarded as positive when two points were present. The neovascular expression rate in the eye tissue was calculated according to the following formula.
[0043]
Neovascularization rate (%)
= (Number of fluorescence leakage positive points / number of photocoagulation points) × 100
The angiogenesis inhibition rate by each test compound was calculated according to the following formula.
[0044]
Angiogenesis inhibition rate (%)
= (1-neovascular expression rate of test compound / control neovascular rate) x 100
The angiogenesis inhibition rate of each test compound calculated as described above is shown in Table 1 (note that the numbers in the table indicate average values of eight eyes).
[0045]
[Table 1]

[0046]
[2] Formulation example injection (100 ml)
Formulation: N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S) -hydroxysuccinyl] -4 '-[tetraethylbis (phosphono) methylimino] -L-phenylalanine-N-cyclopropyl Amide 500mg

N- [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S) -hydroxysuccinyl] -4 '-[tetraethylbis (phosphono) methylimino] -L-phenylalanine-N-cyclopropylamide and sodium chloride were dissolved to obtain 100 ml of an injection.
[0048]
【The invention's effect】
As is clear from the results of the above pharmacological tests, this compound has an excellent angiogenesis inhibitory action in ocular tissues, so that diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein occlusion , Retinal artery occlusion, pterygium, rubeosis, corneal neovascular disease and the like are useful as preventive and therapeutic agents for ophthalmic diseases involving angiogenesis. Further, the present compound has a hydrophilic group and has improved solubility in water, and thus is suitable for aqueous preparations such as injections and eye drops.

Claims (7)

An ocular tissue angiogenesis inhibitor comprising, as an active ingredient, a compound represented by the following general formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof.

[In the formula, R ¹ represents hydrogen, a hydroxyl group, an aryl (C ₁ -C ₆ ) alkylene group or an —A-SOn-B group (A represents a (C ₁ -C ₆ ) alkylene group; A C ₁ -C ₆ ) alkyl group, a (C ₁ -C ₆ ) acyl group, an aryl group or a 5- to 6-membered ring skeleton having at least one sulfur, oxygen or nitrogen heteroatom, wherein n is 0, 1 or R ² represents hydrogen, a (C ₁ -C ₆ ) alkyl group, a (C ₁ -C ₆ ) alkyloxy group or a (C ₁ -C ₆ ) alkylthio group. And R ³ and R ⁴ may be the same or different and represent hydrogen, a (C ₁ -C ₆ ) alkyl group, a (C ₃ -C ₁₀ ) cycloalkyl group, an aryl group or an aryl (C ₁ -C ₆ ) Represents an alkylene group, R ⁵ represents a hydrophilic group, and R ⁶ represents water And represents an unsubstituted benzyl group, a trialkylsilyl group, a tert-butyldiphenylsilyl group, a tetrahydropyranyl group or a tert-butyl group. ] R ^{5 represents} a —Y—C group or a C group (Y represents a (C ₁ -C ₆ ) alkylene group, oxygen, an imino group, or a (C ₁ -C ₆ ) alkyleneimino group; Represents an acid group, a phosphonic acid group, an amidino group, a (C ₁ -C ₆ ) acyl group, an acylimidoyl group, a diphosphonomethine group, a dicarboxymethine group or a tetra (C ₁ -C ₆ ) alkylbisphosphonomethylimino group However, when Y is a (C ₁ -C ₆ ) alkylene group, C is not a (C ₁ -C ₆ ) acyl group.) The ocular tissue angiogenesis inhibitor according to claim 1. R ⁵ is a sulfuric acid group, a sulfonic acid group, a phosphoric acid group, a guanide group, a guanidomethylene group, an amidino group, an acetimidoyliminomethylene group, a propionimidoyliminomethylene group, a benzimidoyliminomethylene group, or acetamide. The ocular tissue angiogenesis inhibitor according to claim 1, which is a methylene group or a tetraethylbisphosphonomethylimino group. In the general formula (I), R ¹ is hydrogen, a hydroxyl group, a phenyltrimethylene group, a thienylthiomethylene group or an isopropylthiomethylene group, R ² is an isopropyl group or an n-propyloxy group, and R ³ Is hydrogen, R ⁴ is a methyl group, a phenyl group, a cyclopropyl group, a p-methoxyphenyl group or a 1-naphthyl group, and R ⁵ is a sulfate group, a sulfonate group, a phosphate group, a guanide group The inhibitor of claim 1, wherein R ⁶ is hydrogen, a guanidomethylene group or a tetraethylbisphosphonomethylimino group, and R ⁶ is hydrogen. N- [4- (N-hydroxyamino) -2-isobutyl-3- (3-phenyltrimethylene) succinyl] -L-4′-guanidephenylalanine-N-methylamide, N- [4- (N-hydroxy Amino) -2-isobutyl-3-hydroxysuccinyl] -L-4′-guanidophenylalanine-N-methylamide, N- [4- (N-hydroxyamino) -2-isobutyl-3- (3-phenyltrimethylene) ) Succinyl] -O-sulfo-L-tyrosine-N-1-naphthylamide or N- [4- (N-hydroxyamino) -2-isobutyl-3-hydroxysuccinyl] -4 '-[tetraethylbis (phosphono) Methylimino] -L-phenylalanine-N-cyclopropylamide, a stereoisomer thereof, or a compound thereof Ocular tissue an angiogenesis inhibitor comprising as an active ingredient a pharmaceutically acceptable salt or solvate thereof. A therapeutic agent for an ophthalmic disease associated with angiogenesis, comprising as an active ingredient the compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof. 7. The therapeutic agent according to claim 6, wherein the ophthalmic disease is diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein occlusion, retinal artery occlusion, pterygium, rubeosis or corneal neovascular disease.