JP4120713B2 - Treatment for multiple sclerosis - Google Patents

Description

【００１０】
［式中Ｒはフェニル環上に低級アルコキシ基を有することのあるベンゾイル基を示す。カルボスチリル骨格の３位と４位との炭素間結合は一重結合又は二重結合を示す。］
【００１１】
【発明の実施の形態】
本発明において有効成分として用いられるカルボスチリル誘導体は、例えば特公平１−４３７４７号公報及び米国特許番号ＵＳ４，４１５，５７２号公報に記載される通り、強心剤として有用であることが既に公知である。該カルボスチリル誘導体及び／又はその塩の中の一つは、また、うっ血性心不全の患者において、血行力学的指標と運動耐用量の改善が証明されている〔Inoue et al., Heart Vessels, 2, 166-171(1986); Sasayma et al., Heart Vessels, 2, 23-28(1986); Feldman et al., Am. Heart J., 116, 771-777(1988)〕。上記カルボスチリル誘導体の変力作用に関連したメカニズムは、カリウム電流の低下〔Iijima et al., J. Pharmacol. Exp. Ther., 240, 657-662(1987)〕、フォスフォジエステラーゼの軽度の抑制とカルシウム電流の内向きの流れの上昇〔Yatani et al., J. Cardiovasc. Pharmacol., 13, 812-819(1989); Taira et al., Arzneimittelforshung, 34, 347-355(1984)〕を含んでいる。
【００１２】
上記カルボスチリル誘導体は、更にリポポリサッカライド（LPS）刺激した末梢血単核球の細胞（PBMC）によるＴＮＦ−α、ＩＬ−６、ＩＬ−１β、ＩＬ−２やＩＦＮ−γを含む種々のサイトカインの産生を抑制することが報告されている〔Busch, F. et al., Eur. J. Clin. Pharmacol., 42, 629-634(1992); Maruyama et al., Biochem. Biophys. Res. Commu., 195, 1264-1271(1993); Shioi, T. et al., Life Sciences, 54(1), PL11-16(1994); Matsumori, A. et al., Circulation, 89(3), 955-958(1994)〕。
【００１３】
更に上記カルボスチリル誘導体は、ウイルス誘導された心筋傷害の改善と該心筋傷害の改善時におけるナチュラル・キラー細胞(NK Cell)の抑制作用を有することも報告されている〔Matsui S., et al., J. Clin. Invest., 94, 1212-1217(1994)〕。上記作用に加えて、該カルボスチリル誘導体は、アポトーシス調整作用を有することも報告され、該作用に基づく、癌の治療及び癌転移の抑制、自己免疫疾患、ウイルス疾患等への有用性も報告されている〔ヨーロッパ特許公開EP0552373; Nakai et al., Jpn. J. Cancer Res., abstruct, and Proc. Jpn. Cancer Assoc., 581(1993); Maruyama et al., Biochem. Biophys. Res. Commun., 195, 1264-1271(1993)〕。
【００１４】
しかして、上記一般式（１）で表わされるカルボスチリル誘導体及び／又はその塩が、前記したように多発性硬化症の臨床症状の抑制作用を有する報告は、未だない。
【００１５】
本発明の多発性硬化症治療剤において、殊に好ましい上記カルボスチリル誘導体としては、６−［４−（３，４−ジメトキシベンゾイル）−１−ピペラジニル］−３，４−ジヒドロカルボスチリルを例示できる。
【００１６】
本発明薬剤の有効成分とするカルボスチリル誘導体を表わす上記一般式（１）において示される各基としては、より具体的にはそれぞれ次の各基を例示できる。
【００１７】
即ち、低級アルコキシ基としては、例えばメトキシ、エトキシ、プロポキシ、イソプロポキシ、ブトキシ、tert−ブトキシ、ペンチルオキシ、ヘキシルオキシ基等の炭素数１〜６の直鎖又は分枝鎖状アルコキシ基を例示することができる。
【００１８】
フェニル環上に置換基として低級アルコキシ基を有することのあるベンゾイル基としては、例えばベンゾイル、２−メトキシベンゾイル、３−メトキシベンゾイル、４−メトキシベンゾイル、２−エトキシベンゾイル、３−エトキシベンゾイル、４−エトキシベンゾイル、３−イソプロポキシベンゾイル、４−ブトキシベンゾイル、２−ペンチルオキシベンゾイル、３−ヘキシルオキシベンゾイル、３，４−ジメトキシベンゾイル、２，５−ジメトキシベンゾイル、３，４，５−トリメトキシベンゾイル基等の、フェニル環上に置換基として炭素数１〜６の直鎖又は分枝鎖状アルコキシ基の１〜３個を有することのあるベンゾイル基を例示することができる。
【００１９】
また上記一般式（１）で表されるカルボスチリル誘導体の塩には、薬理学的に許容される酸付加塩が包含される。該塩を形成する酸性化合物としては、具体的には例えば硫酸、リン酸、硝酸、塩酸、臭化水素酸等の無機酸、蓚酸、マレイン酸、フマール酸、リンゴ酸、酒石酸、クエン酸、安息香酸等の有機酸を例示することができる。
【００２０】
本発明薬剤の有効成分である一般式（１）で表わされるカルボスチリル誘導体及び／又はその塩は、通常、一般的な医薬製剤の形態で用いられる。かかる製剤は、通常使用される充填剤、増量剤、結合剤、付湿剤、崩壊剤、表面活性剤、潤沢剤等の希釈剤乃至賦形剤を用いて調製される。この医薬製剤としては各種の形態が治療目的に応じて選択でき、その代表的なものとしては錠剤、丸剤、散剤、液剤、懸濁剤、乳剤、顆粒剤、カプセル剤、坐剤、注射剤（液剤、懸濁剤等）、点眼剤等を例示できる。
【００２１】
錠剤の形態に形成するに際しては、担体としてこの分野で従来公知のものを広く使用でき、例えば乳糖、白糖、塩化ナトリウム、ブドウ糖、尿素、デンプン、炭酸カルシウム、カオリン、結晶セルロース、ケイ酸等の賦形剤、水、エタノール、プロパノール、単シロップ、ブドウ糖液、デンプン液、ゼラチン溶液、カルボキシメチルセルロース、セラック、メチルセルロース、リン酸カリウム、ポリビニルピロリドン等の結合剤、乾燥デンプン、アルギン酸ナトリウム、カンテン末、ラミナラン末、炭酸水素ナトリウム、炭酸カルシウム、ポリオキシエチレンソルビタン脂肪酸エステル類、ラウリル硫酸ナトリウム、ステアリン酸モノグリセリド、デンプン、乳糖等の崩壊剤、白糖、ステアリン、カカオバター、水素添加油等の崩壊抑制剤、第４級アンモニウム塩基、ラウリル硫酸ナトリウム等の吸収促進剤、グリセリン、デンプン等の保湿剤、デンプン、乳糖、カオリン、ベントナイト、コロイド状ケイ酸等の吸着剤、精製タルク、ステアリン酸塩、ホウ酸末、ポリエチレングリコール等の滑沢剤等を例示できる。更に錠剤は必要に応じて通常の剤皮を施した錠剤、例えば糖衣錠、ゼラチン被包錠、腸溶被錠、フィルムコーティング錠あるいは二重錠、多層錠とすることもできる。
【００２２】
丸剤の形態に成形するに際しては、担体としてこの分野で従来公知のものを広く使用でき、例えばブドウ糖、乳糖、デンプン、カカオ脂、硬化植物油、カオリン、タルク等の賦形剤、アラビアゴム末、トラガント末、ゼラチン、エタノール等の結合剤、ラミナランカンテン等の崩壊剤等を使用できる。
【００２３】
坐剤の形態に成形するに際しては、担体として従来公知のものを広く使用でき、例えばポリエチレングリコール、カカオ脂、高級アルコール、高級アルコールのエステル類、ゼラチン、半合成グリセライド等を使用できる。
【００２４】
注射剤として調製される場合には、液剤、乳剤、懸濁剤などは殺菌され且つ血液と等張であるのが好ましく、これらの液剤、乳剤、懸濁剤などの形態に成形するに際しては、希釈剤としてこの分野において慣用されているものを全て使用できる。その例としては例えば水、エチルアルコール、プロピレングリコール、エトキシ化イソステアリルアルコール、ポリオキシ化イソステアリルアルコール、ポリオキシエチレンソルビタン脂肪酸エステル類等を挙げることができる。尚、この場合等張性の溶液を調製するに充分な量の食塩、ブドウ糖、グリセリン等を医薬製剤中に含有させてもよく、また通常の溶解補助剤、緩衝剤、無痛化剤等を添加してもよい。更に必要に応じて着色剤、保存剤、香料、風味剤、甘味剤等や他の医薬品を医薬製剤中に含有させてもよい。
【００２５】
本発明薬剤中に有効成分として含まれる一般式（１）の化合物の量は、特に限定されず広範囲より適宜選択されるが、通常全組成物中約１〜７０重量％、好ましくは約１〜３０重量％程度の範囲とするのが適当である。
【００２６】
かくして得られる本発明多発性硬化症治療剤の投与方法は特に制限はなく、各種製剤形態、患者の年齢、性別、その他の条件、疾患の程度に応じて決定される。例えば、注射剤形態の医薬製剤は、静脈内、筋肉内、皮下、皮内、腹腔内投与などにより投与され得る、これは必要に応じてブドウ糖、アミノ酸等の通常の補液と混合して静脈内投与することもできる。錠剤、丸剤、顆粒剤、カプセル剤などの固剤形態や経口投与用液剤形態の本発明医薬製剤は、経口投与又は経腸投与され得る。坐剤は直腸内投与できる。また点眼剤は目に適用できる。
【００２７】
本発明薬剤の投与量は、広範囲から適宜選択でき、特に限定されるものではないが、通常の臨床利用に際しては、一般式（１）のカルボスチリル誘導体（及びその塩）が、１日当り体重１ｋｇ当り約０．５〜３０ｍｇ程度となる量の範囲から選択されるのがよく、前記投与単位形態中には有効成分が約１〜１０００ｍｇ含有されるのが適当である。また本発明薬剤の投与は、１日１回又は１日３〜４回に分けることもできる。
【００２８】
【発明の効果】
本発明の多発性硬化症治療剤は、多発性硬化症の各種臨床症状の改善や、多発性硬化症の再発予防及び治療に有効である。
【００２９】
【実施例】
以下、本発明を更に詳しく説明するため製剤例及び薬理試験例を挙げる。
【００３０】
【製剤例１】

１錠中、上記組成物の錠剤を製造した。
【００３１】
【製剤例２】
６−〔４−（３，４−ジメトキシベンゾイル）−１−ピペラジニル〕−３，４−ジヒドロカルボスチリル １５０ｇ
アビセル（商標名、旭化成株式会社製） ４０ｇ
コーンスターチ ３０ｇ
ステアリン酸マグネシウム ２ｇ
ヒドロキシプロピルメチルセルロース １０ｇ
ポリエチレングリコール−６０００ ３ｇ
ヒマシ油 ４０ｇ
メタノール ４０ｇ
上記有効成分化合物、アビセル、コーンスターチ及びステアリン酸マグネシウムを混合研磨後、糖衣Ｒ１０ｍｍのキネで打錠する。得られた錠剤をヒドロキシプロピルメチルセルロース、ポリエチレングリコール−６０００、ヒマシ油及びメタノールからなるフィルムコーティング剤で被覆を行ない、フィルムコーティング錠を製造した。
【００３２】
【製剤例３】
６−〔４−（３，４−ジメトキシベンゾイル）−１−ピペラジニル〕−３，４−ジヒドロカルボスチリル １５０．０ｇ
クエン酸 １．０ｇ
ラクトース ３３．５ｇ
リン酸二カルシウム ７０．０ｇ
プルロニックＦ−６８ ３０．０ｇ
ラウリル硫酸ナトリウム １５．０ｇ
ポリビニルピロリドン １５．０ｇ
ポリエチレングリコール（カルボワックス１５００） ４．５ｇ
ポリエチレングリコール（カルボワックス６０００） ４５．０ｇ
コーンスターチ ３０．０ｇ
乾燥ラウリル硫酸ナトリウム ３．０ｇ
乾燥スタアリン酸マグネシウム ３．０ｇ
エタノール 適 量
上記有効成分化合物、クエン酸、ラクトース、リン酸二カルシウム、プルロニックＦ−６８及びラウリル硫酸ナトリウムを混合した。
【００３３】
上記混合物をＮｏ．６０スクリーンにて篩別し、ポリビニルピロリドン、カルボワックス１５００及びカルボワックス６０００を含むアルコール性溶液で湿式粒状化した。必要に応じてアルコールを添加し、粉末をペースト状塊にした。コーンスターチを添加し、均一な粒子が形成されるまで混合を続けた。Ｎｏ．１０スクリーンを通過させ、トレイに入れ、１００℃のオーブンで１２〜１４時間乾燥し、乾燥粒子をＮｏ．１６スクリーンで篩別し乾燥ラウリル硫酸ナトリウム及び乾燥ステアリン酸マグネシウムを加え、混合し、打錠機で所望の形状に圧縮成形した。
【００３４】
上記芯部をワニスで処理し、タルクを散布して湿気の吸収を防止した。芯部の周囲に下塗り層を被覆した。内服用のために充分な回数のワニス被覆を行なった。錠剤を完全に丸く且つ滑らかにするために、更に下塗り層及び平滑被覆を適用した。所望の色合いが得られるまで着色被覆を行ない、乾燥後、被覆錠剤を磨いて均一な光沢の錠剤を調製した。
【００３５】
【薬理試験例１】
供試化合物として６−［４−（３，４−ジメトキシベンゾイル）−１−ピペラジニル］−３，４−ジヒドロカルボスチリル（以下「化合物１」という）を用いて、以下の通り、多発性硬化症の動物モデルとされている実験的アレルギー性脳脊髄炎（experimental allergic encephalomyelitis;ＥＡＥ：Paterson,P.Y., Fed. Proc., 41, 2569-2576(1982); Paterson, P.Y.,Immunological Diseases, 3rd edn., Little, Brown & Co., Boston, MA. pp.1400-1435(1978); Alvord,E.C.Jr., Progress in Clinical and Biological Research, 148. Alan R. Liss, New York, NY, pp.523-537(1984)）を作製し、臨床スコアに対する供試化合物の抑制効果をコォーら（Koh, C.S.,et al., Cell Immunology, 107, 52-63(1987)）の方法に準じて評価した。
【００３６】
上記ＥＡＥ動物モデルは、動物に中枢神経組織（例えば、モルモット脊髄）とアジュバントで感作することによりＥＡＥを引き起こすことができる。まず、免疫後９−１０日目に尾及び下肢の麻痺などを伴った症状が発現し、その後、モルモット脊髄（Guinea-Pig Spinal Cord; GPSC）ホモジネートと高用量の結核死菌（Mycobacterium Tuberculosis; MT）を含むアジュバントを用いた場合は、免疫後約４０日間にわたり２−３回の発症、回復を繰り返す慢性進行型の多発性硬化症に対するモデルとなるとされている（Chabannes, D., et al., J. Autoimmunity, 5, 199-211(1992)）。
【００３７】
（１）動物及び実験材料の調製
１）コントロールとして０．５％−ＣＭＣ溶液（カルボキシ・メチル・セルロース溶液）を用いた。該溶液の調製は、ＣＭＣ（セロゲンＦ−ＳＢ(ロット番号１５４４５５)、第一工業製薬社製）を電子天秤（ＥＲ−１８２Ａ；ザルトリウス社製）で５．０ｇ秤量後、メスシリンダーに移し、注射用蒸留水（大塚製薬工場社製）にて４℃で一昼夜撹袢溶解し、最終濃度０．５％になるように調製した後、オートクレーブで滅菌後、使用まで４℃にて保存した。
【００３８】
２）化合物１（ロット番号５Ｄ９６Ｍ）の原末を秤量後、メノウ鉢に移し、少量の０．５％ＣＭＣ溶液で均一に懸濁した後、目的とする濃度になるよう０．５％ＣＭＣ溶液にて希釈調製し、化合物１の最終濃度が６０ｍｇ／ｍｌの濃度になるよう調製し、滅菌遠心チューブに分注し、使用まで４℃にて遮光保存した。
【００３９】
３）免疫原として用いるモルモット脊髄（ＧＰＳＣ）ホモジネートは、９−１０週齢（５００−６００ｇ）の雄性ハートレー系モルモット（日本チヤールズリバー社製より購入）２０匹全匹を使用した。モルモットを軽エーテル麻酔下に頚椎脱臼により安楽死させ、脊髄から髄膜と血管を剥離し、氷冷したシャーレに摘出した脊髄を移した。摘出した脊髄１０匹分位の量をまとめて秤量し、ポッター型ホモジナイザー中にＧＰＳＣ１ｇに対して１ｍｌの冷生理食塩水（大塚製薬工場社製）を加え、ホモジネート調製用ローター（池本理科工業社製）を用いて、氷冷下でホモジネートを作製した（ＧＰＳＣ濃度として５００ｍｇ／ｍｌ）。３〜４ｍｌずつＰＰＰチューブに分注し、−２０℃にて凍結保存した。
【００４０】
４）結核死菌（ＭＴ）アジュバントの調製は、市販の結核死菌含有完全アジュバント溶液（ＭＴ，Ｈ３７Ｒａ濃度１ｍｇ／ｍｌ；カタログ番号３１１３−６０−５：ディフコ社製）１０ｍｌ／瓶に、更に結核死菌Ｈ３７Ｒａ(カタログ番号３１１４−３３−８:ディフコ社製)を１５ｍｇ添加し、パラフィルムでシーリング後よく転倒混和した後、均一に懸濁させ、最終ＭＴ濃度２．５ｍｇ／ｍｌに調製した。
【００４１】
５）上記３）及び４）で調製したＧＰＳＣホモジネートと結核死菌含有アジュバント溶液を、三方活栓に接続した２本のガラスシリンジに１：１の割合で移し、均一なエマルジョンが形成されるまで繰り返し混和して調製し、得られたエマルジョン溶液を免疫エマルジョンとして使用した。
【００４２】
６）実験のための動物として、雌性ルイス（Lewis）系ラット（ＳＰＦ：日本チャールズリバー社より購入）の１０週齢を使用した。９週齢で１００匹購入し、１週間飼育馴化後、体重を指標に層別無作為抽出により、各群２０匹／４ケージＸ３群の全６０匹を本試験のために抽出した。
【００４３】
（２）多発性硬化症の動物モデルの作成
雌性ルイス系ラット体重１８０−２００ｇの１０週齢を試験に供した。ラットには水及び普通飼料を経口にて自由摂取させた。
【００４４】
多発性硬化症の動物モデルとしての実験的アレルギー性脳脊髄炎（ＥＡＥ）の作成は、次の通り行なった。即ち、上記（１）−５）で調製した免疫エマルジョンを軽エーテル麻酔下のラット両足フットパッドに２３ゲージの注射針と１．０ｍｌのガラスシリンジを用いて０．１ｍｌずつ合計０．２ｍｌ、ＧＰＳＣとして５０ｍｇ＋ＭＴとして２５０μｇ／体、皮内注射した。免疫後、飼育は５匹／ケージで行なった。該ラットは免疫後、１０日前後にてアレルギー性脳脊髄炎症状が発症し、ヒト臨床症状と同じく、症状の再発と寛解が繰り返され、従って多発性硬化症の動物モデルとなる。
【００４５】
（３）本発明化合物１の多発性硬化症治療効果及び再発予防効果の検討
１） （１）−２）で調製した本発明化合物１の懸濁液をラットへ投与する直前にチューブをよく転倒混和した後、１ｍｌ又は２．５ｍｌのテルモシリンジに取り、ラット用経口ゾンデを用いて、ラットの体重１００ｇ当たり０．５ｍｌを強制経口投与した。尚、投与量は、コンピューターに自動入力した当日の体重によって１匹毎に決定した。コントロール群としての０．５％ＣＭＣ溶媒投与群も同様な方法で投与した。
【００４６】
多発性硬化症の動物モデルに対する実験は、コントロール群(ＣＭＣ溶媒群)、免疫後１−９日目まではＣＭＣ溶媒を投与し、ＥＡＥ発症日の１０日目より化合物１の３００ｍｇ／ｋｇ／日を投与した群（第１の症状発症直後より投与した群）及び免疫後１−１６日目まではＣＭＣ溶媒を投与し、症状が一旦回復後、再び悪化し始める頃の１７日目より化合物１の３００ｍｇ／ｋｇ／日を投与した群（第２の症状発症直前より投与した群）の３群にて、１群それぞれ２０匹を用いて、免疫後３７日まで、１日１回連日経口投与を行なった。
【００４７】
２） 体重、血球測定、胸腺重量、脾臓重量測定及び症状観察(スコア判定)
試験開始日（免疫日）から試験終了日（解剖日）まで、１日１回体重と症状の観察(スコア判定)を行なった。体重は、電子天秤（Sartorius; LC-4200:ザルトリウス社製）にて測定し、接続したパーソナル・コンピュータ（TOSHIBA J3100:東芝製）に自動入力し、同時に個体毎の投与量も決定表示させた。
【００４８】
症状のスコア判定は、コォー（Koh）らの評価基準（Koh, C.-S., et al., Cell Immunology, 107, 52-63 (1987)）に従って、毎日各群のラットの症状を観察し、ラット尾部と両後肢のそれぞれに対してスコア判定し、その合計をスコア記録シートに記載した。スコア判定は、尾部については、無症状を０、途中まで弛緩を０．２５、尾部全体の完全な弛緩を０．５とし、また後肢については、無症状を０、歩行障害を１．０、両後肢の不完全麻痺及び片側後肢の完全麻痺を２．０及び両後肢の完全麻痺を３．０として評価した。尚、試験中の死亡例はスコア判定の統計処理から除外した。
【００４９】
３） 統計処理法
全ての統計学的分析は、ＳＡＳシステム(SAS Institute Japan, Ver.6.11)により行なった。
【００５０】
体重は、コントロール群を対照として、第一アタック以降化合物１投与群、第二アタック以降化合物１投与群について、実測値を用い、繰り返し測定に基づく分散分析(Repeated Mesurements)により経時的な反応の違いを検定し、有意水準を５％及び１％で実施した。
【００５１】
症状観察(症状のスコア判定)
試験期間全体をコントロール群の臨床スコアの推移によって、前半と後半の２つに分割した。即ち、スコアの上で２回のピーク（Chabannes, D., et al., J. Autoimmunity, 5, 199-211 (1992)）の間の一番回復した時点で、前半と後半に分け、全期間、前半、後半の３つの期間で解析を行なった。より具体的には、それぞれの期間における各個体の症状のスコア判定結果を積算し、各群において平均積算臨床スコア(Integrated Clinical Score)を求め（Branisteanu, D.D., et al., J. Neuroimmunology, 61, 151-160 (1995); Martin, D., et al., J. Neuroimmunology, 61, 241-245 (1995)）、一元配置分散分析後、コントロール群を対照として両側ダネット−テスト(Dunnett-test)にて検定し、有意水準を５％及び１％で実施した。尚、スコアは、平均スコアと標準誤差で表わた。
【００５２】
血球測定、胸腺重量及び脾臓重量については、一元配置分散分析後、コントロール群を対照として両側ダネット−テストにて検定し、有意水準を５％及び１％で実施した。
【００５３】
症状のスコア判定結果と体重の推移を図１、図２及び図３にそれぞれ示す。
【００５４】
図１において、縦軸は、平均臨床スコアを、横軸は免疫後の投与期間（日）を示し、（１）はコントロール(ＣＭＣ溶媒のみ)群の結果、（２）は化合物１の３００mg/kg/日を免疫後１７日目より投与開始した群の結果、（３）は化合物１の３００mg/kg/日を免疫後１０日目より投与開始した群の結果を、それぞれ示す。
【００５５】
図２において、縦軸は平均積算臨床スコア（０−３８日までの臨床スコアの積算の平均）を、横軸は（１）：コントロール(ＣＭＣ溶媒のみ)群、（２）：化合物１の３００mg/kg/日を免疫後１７日目より投与開始した群及び（３）：化合物１の３００mg/kg/日を免疫後１０日目より投与開始した群を、それぞれ示す。
【００５６】
図３において、縦軸は毎日の体重の推移を、横軸は（１）：コントロール(ＣＭＣ溶媒のみ)群、（２）化合物１の３００mg/kg/日を免疫後１７日目より投与開始した群及び（３）：化合物１の３００mg/kg/日を免疫後１０日目より投与開始した群を、それぞれ示す。
【００５７】
上記の結果から、次のことが明らかである。即ち、図２から確認されるように化合物１投与群はコントロール群と比較して、いずれも平均積算臨床スコアの有意な低下(P<0.01)、即ち症状の改善効果が観察された。また、体重の推移は、図３に示されるように、図１における症状のスコア判定の推移とよく対応して、化合物１投与群では、コントロール群と比較して、改善傾向が認められた。更に、図１に示されるように、化合物１は、第１の症状発症直後の投与によりその症状を改善し、第２の症状の発症を抑制していることが分かる。また第２の症状発症直前より化合物１を投与した群は、その症状の再発を抑制していることから、臨床において治療困難な再発・回復型及び慢性進行性多発性硬化症の再発予防及び各種臨床症状に対する治療効果が期待できることが判る。
【図面の簡単な説明】
【図１】薬理試験例１に従う試験の結果を示すグラフである。
【図２】薬理試験例１に従う試験の結果を示すグラフである。
【図３】薬理試験例１に従う試験の結果を示すグラフである。[0001]
[Industrial application fields]
The present invention relates to a therapeutic agent for multiple sclerosis comprising a pharmaceutically useful carbostyril derivative represented by the following general formula (1) and / or a salt thereof as an active ingredient.
[0002]
[Prior art]
Multiple sclerosis is a demyelinating disease of the central nervous system of unknown cause, mainly affecting young adults, and multifocal demyelinating lesions occur in the central nervous tissue such as the brain, spinal cord, and optic nerve, It is a progressive intractable neurological disease characterized by a variety of temporal and spatial multiple occurrences of various neurological symptoms that recur and relapse.
Clinical symptoms can be any disorder of the central nervous system, but include visual impairment based on optic nerve and spinal cord disorders, motor paralysis, gait disturbance, numbness, abnormal sensation, sensory paralysis, and eye pain. In other cases, brainstem disorders may cause double vision, and cerebellar lesions may cause ataxia. The most common age is 20-40 years old. The etiology of the multiple sclerosis is unknown until now, and there are autoimmune theory and virus theory, but neither theory has become definitive.
[0003]
The diagnosis of the multiple sclerosis is based on clinical characteristics mainly because the etiology is unknown as described above and there is no specific examination abnormality. The diagnostic criteria that are often used at present are the diagnostic criteria for multiple sclerosis (MS) by the research group of the “Multiple sclerosis” specific disease of the Ministry of Health and Welfare, and the MS diagnostic criteria for Poser, Poser, CM, (1983), etc. It has been known. Also, recently, is a useful diagnosis by MRI, as the findings of this disease, the limbic oval in T ₂ -weighted images is relatively clear, high intensity area of the lesion (plaque) are found in the cerebral white matter and brain stem That's it. .
[0004]
Treatment of multiple sclerosis can be divided into treatments for etiology and symptomatic treatments for various central nervous system symptoms. Since the etiology of the former is unknown at present, immunosuppressive drugs and anti-inflammatory drugs are used for the treatment, but it is difficult to determine the effect. In the acute phase, corticosteroids are used, and when the symptoms are more acute and severe, steroid pulse therapy is administered. Chronic treatment is mainly symptomatic treatment and rehabilitation for neuropathy.
[0005]
Recently, treatment with interferon-β (IFN-β) has been reported to reduce the number of recurrences of multiple sclerosis as a preventive therapy / refractory treatment of recurrence, and IFN-β is used for treatment in the United States. However, the treatment method for multiple sclerosis has not been established yet, and the development of a drug having a therapeutic effect on the prevention of recurrence of the multiple sclerosis and various clinical symptoms is desired in this field.
[0006]
[Problems to be solved by the invention]
Accordingly, an object of the present invention is to develop a drug having a therapeutic effect on the prevention of relapse of multiple sclerosis and various clinical symptoms desired in the field.
[0007]
[Means for Solving the Problems]
As a result of intensive research for the above purpose, the present inventor has successfully developed a carbostyril derivative represented by the following general formula (1) and / or a salt thereof as a therapeutic agent for multiple sclerosis. As a result, the present invention has been completed.
[0008]
That is, the present invention relates to a therapeutic agent for multiple sclerosis comprising a carbostyril derivative represented by the following general formula (1) and / or a salt thereof as an active ingredient, in particular, the above carbostyril derivative is 6- [4- (3,4). The present invention provides a therapeutic agent for multiple sclerosis, which is -dimethoxybenzoyl) -1-piperazinyl] -3,4-dihydrocarbostyril.
[0009]
[Chemical 2]

[0010]
[Wherein R represents a benzoyl group which may have a lower alkoxy group on the phenyl ring. The carbon-carbon bond at the 3-position and 4-position of the carbostyryl skeleton represents a single bond or a double bond. ]
[0011]
DETAILED DESCRIPTION OF THE INVENTION
The carbostyril derivative used as an active ingredient in the present invention is already known to be useful as a cardiotonic agent as described in, for example, JP-B-1-43747 and US Pat. No. 4,415,572. One of the carbostyril derivatives and / or salts thereof, also in patients with congestive heart failure, improving the hemodynamic indices motion tolerated dose has been demonstrated [Inoue et al., Heart Vessels, 2 , 166-171 (1986); Sasayma et al., Heart Vessels, 2 , 23-28 (1986); Feldman et al., Am. Heart J., 116 , 771-777 (1988)]. The mechanism related to the inotropic action of the above-mentioned carbostyril derivatives is the decrease in potassium current (Iijima et al., J. Pharmacol. Exp. Ther., 240 , 657-662 (1987)), the mildness of phosphodiesterase. Inhibition and increased inward flow of calcium current (Yatani et al., J. Cardiovasc. Pharmacol., 13 , 812-819 (1989); Taira et al., Arzneimittelforshung, 34 , 347-355 (1984)) Contains.
[0012]
The above carbostyril derivatives are various cytokines including TNF-α, IL-6, IL-1β, IL-2 and IFN-γ by peripheral blood mononuclear cells (PBMC) stimulated with lipopolysaccharide (LPS). [Busch, F. et al., Eur. J. Clin. Pharmacol., 42 , 629-634 (1992); Maruyama et al., Biochem. Biophys. Res. Commu ., 195 , 1264-1271 (1993); Shioi, T. et al., Life Sciences, 54 (1), PL11-16 (1994); Matsumori, A. et al., Circulation, 89 (3), 955 -958 (1994)].
[0013]
Furthermore, the carbostyril derivatives have been reported to have an effect of improving virus-induced myocardial damage and suppressing natural killer cells (NK cells) when the myocardial damage is improved [Matsui S., et al. , J. Clin. Invest., 94 , 1212-1217 (1994)]. In addition to the above effects, the carbostyril derivatives are also reported to have an apoptosis regulating action, and based on these actions, usefulness for cancer treatment and suppression of cancer metastasis, autoimmune diseases, viral diseases, etc. has also been reported. [European Patent Publication EP0552373; Nakai et al., Jpn. J. Cancer Res., Abstruct, and Proc. Jpn. Cancer Assoc., 581 (1993); Maruyama et al., Biochem. Biophys. Res. Commun. , 195, 1264-1271 (1993)].
[0014]
As described above, there is still no report that the carbostyril derivative represented by the general formula (1) and / or a salt thereof has an action of suppressing clinical symptoms of multiple sclerosis.
[0015]
In the therapeutic agent for multiple sclerosis of the present invention, particularly preferable carbostyril derivatives include 6- [4- (3,4-dimethoxybenzoyl) -1-piperazinyl] -3,4-dihydrocarbostyril. .
[0016]
As each group shown in the said General formula (1) showing the carbostyril derivative used as an active ingredient of this invention chemical | medical agent, the following each group can be illustrated more specifically, respectively.
[0017]
That is, examples of the lower alkoxy group include linear or branched alkoxy groups having 1 to 6 carbon atoms such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentyloxy, and hexyloxy groups. be able to.
[0018]
Examples of the benzoyl group which may have a lower alkoxy group as a substituent on the phenyl ring include benzoyl, 2-methoxybenzoyl, 3-methoxybenzoyl, 4-methoxybenzoyl, 2-ethoxybenzoyl, 3-ethoxybenzoyl, 4- Ethoxybenzoyl, 3-isopropoxybenzoyl, 4-butoxybenzoyl, 2-pentyloxybenzoyl, 3-hexyloxybenzoyl, 3,4-dimethoxybenzoyl, 2,5-dimethoxybenzoyl, 3,4,5-trimethoxybenzoyl group Examples thereof include a benzoyl group having 1 to 3 linear or branched alkoxy groups having 1 to 6 carbon atoms as substituents on the phenyl ring.
[0019]
The salt of the carbostyril derivative represented by the general formula (1) includes a pharmacologically acceptable acid addition salt. Specific examples of the acidic compound that forms the salt include inorganic acids such as sulfuric acid, phosphoric acid, nitric acid, hydrochloric acid, and hydrobromic acid, oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, and benzoic acid. An organic acid such as an acid can be exemplified.
[0020]
The carbostyril derivative represented by the general formula (1) and / or a salt thereof, which is an active ingredient of the drug of the present invention, is usually used in the form of a general pharmaceutical preparation. Such preparations are prepared using diluents or excipients such as fillers, extenders, binders, moisturizers, disintegrants, surfactants, lubricants and the like that are usually used. Various forms of this pharmaceutical preparation can be selected according to the purpose of treatment, and typical ones are tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, injections. (Liquids, suspensions, etc.), eye drops and the like.
[0021]
In forming a tablet, conventionally known carriers can be widely used as carriers, such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid and the like. Form, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone and other binders, dry starch, sodium alginate, agar powder, laminaran powder , Disintegrating agents such as sodium bicarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose, disintegration inhibitors such as sucrose, stearin, cocoa butter, hydrogenated oil, Absorption promoters such as quaternary ammonium base and sodium lauryl sulfate, humectants such as glycerin and starch, adsorbents such as starch, lactose, kaolin, bentonite and colloidal silicic acid, purified talc, stearate, boric acid powder, Examples thereof include lubricants such as polyethylene glycol. Furthermore, the tablet can be a tablet coated with a normal coating as required, for example, a sugar-coated tablet, a gelatin-encapsulated tablet, an enteric-coated tablet, a film-coated tablet, a double tablet, or a multilayer tablet.
[0022]
In molding into a pill form, conventionally known carriers can be widely used as carriers, such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, talc and other excipients, gum arabic powder, A binder such as tragacanth powder, gelatin, ethanol or the like, a disintegrant such as lamina lankanten or the like can be used.
[0023]
In molding into a suppository, conventionally known carriers can be widely used, for example, polyethylene glycol, cocoa butter, higher alcohols, higher alcohol esters, gelatin, semi-synthetic glycerides, and the like.
[0024]
When prepared as an injection, liquids, emulsions, suspensions and the like are preferably sterilized and isotonic with blood, and when molded into these liquids, emulsions, suspensions and the like, Any of those conventionally used in this field can be used as a diluent. Examples thereof include water, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters and the like. In this case, a sufficient amount of sodium chloride, glucose, glycerin, etc. may be included in the pharmaceutical preparation to prepare an isotonic solution, and usual solubilizing agents, buffers, soothing agents, etc. are added. May be. Furthermore, if necessary, colorants, preservatives, fragrances, flavoring agents, sweetening agents, and other pharmaceuticals may be contained in the pharmaceutical preparation.
[0025]
The amount of the compound of the general formula (1) contained as an active ingredient in the drug of the present invention is not particularly limited and is appropriately selected from a wide range, but is usually about 1 to 70% by weight in the total composition, preferably about 1 to A range of about 30% by weight is appropriate.
[0026]
The administration method of the therapeutic agent for multiple sclerosis of the present invention thus obtained is not particularly limited, and is determined according to various preparation forms, patient age, sex, other conditions, and degree of disease. For example, pharmaceutical preparations in the form of injections can be administered intravenously, intramuscularly, subcutaneously, intradermally, intraperitoneally, etc., which can be intravenously mixed with normal fluids such as glucose and amino acids as necessary. It can also be administered. The pharmaceutical preparation of the present invention in the form of solid preparations such as tablets, pills, granules and capsules or liquid forms for oral administration can be administered orally or enterally. Suppositories can be administered rectally. Eye drops can be applied to the eyes.
[0027]
The dose of the drug of the present invention can be appropriately selected from a wide range, and is not particularly limited. However, for normal clinical use, the carbostyril derivative of general formula (1) (and its salt) is 1 kg body weight per day. The amount is preferably selected from the range of about 0.5 to 30 mg per unit, and it is appropriate that the dosage unit form contains about 1 to 1000 mg of the active ingredient. The administration of the drug of the present invention can be divided once a day or 3 to 4 times a day.
[0028]
【The invention's effect】
The therapeutic agent for multiple sclerosis of the present invention is effective in improving various clinical symptoms of multiple sclerosis and preventing and treating the recurrence of multiple sclerosis.
[0029]
【Example】
Hereinafter, formulation examples and pharmacological test examples are given in order to explain the present invention in more detail.
[0030]
[Formulation Example 1]

In 1 tablet, the tablet of the said composition was manufactured.
[0031]
[Formulation Example 2]
150 g of 6- [4- (3,4-dimethoxybenzoyl) -1-piperazinyl] -3,4-dihydrocarbostyril
Avicel (trade name, manufactured by Asahi Kasei Corporation) 40g
Corn starch 30g
Magnesium stearate 2g
Hydroxypropyl methylcellulose 10g
Polyethylene glycol-6000 3g
Castor oil 40g
40g of methanol
The active ingredient compound, Avicel, corn starch and magnesium stearate are mixed and polished, and then tableted with a sugar-coated R10 mm kine. The obtained tablets were coated with a film coating agent consisting of hydroxypropylmethylcellulose, polyethylene glycol-6000, castor oil, and methanol to produce film-coated tablets.
[0032]
[Formulation Example 3]
6- [4- (3,4-Dimethoxybenzoyl) -1-piperazinyl] -3,4-dihydrocarbostyril 150.0 g
Citric acid 1.0g
Lactose 33.5g
Dicalcium phosphate 70.0g
Pluronic F-68 30.0g
Sodium lauryl sulfate 15.0g
Polyvinylpyrrolidone 15.0g
Polyethylene glycol (Carbowax 1500) 4.5g
Polyethylene glycol (Carbowax 6000) 45.0g
Cornstarch 30.0g
Dry sodium lauryl sulfate 3.0g
Dry magnesium stearate 3.0g
Ethanol appropriate amount The active compound, citric acid, lactose, dicalcium phosphate, pluronic F-68 and sodium lauryl sulfate were mixed.
[0033]
The above mixture was designated as No. The mixture was sieved with 60 screens and wet granulated with an alcoholic solution containing polyvinylpyrrolidone, carbowax 1500 and carbowax 6000. Alcohol was added as needed to make the powder into a pasty mass. Corn starch was added and mixing continued until uniform particles were formed. No. 10 screens, put in a tray, and dried in an oven at 100 ° C. for 12-14 hours. The mixture was sieved with 16 screens, dried sodium lauryl sulfate and dried magnesium stearate were added, mixed, and compressed into a desired shape by a tableting machine.
[0034]
The core was treated with varnish and talc was sprayed to prevent moisture absorption. An undercoat layer was coated around the core. A sufficient number of varnish coatings were performed for internal use. In order to make the tablet completely round and smooth, a further primer layer and a smooth coating were applied. Color coating was performed until the desired shade was obtained, and after drying, the coated tablets were polished to prepare uniform gloss tablets.
[0035]
[Pharmacological Test Example 1]
Using 6- [4- (3,4-dimethoxybenzoyl) -1-piperazinyl] -3,4-dihydrocarbostyril (hereinafter referred to as “Compound 1”) as a test compound, multiple sclerosis is as follows. Experimental allergic encephalomyelitis (EAE): Paterson, PY, Fed. Proc., 41 , 2569-2576 (1982); Paterson, PY, Immunological Diseases, 3rd edn., Little, Brown & Co., Boston, MA. Pp. 1400-1435 (1978); Alvord, ECJr., Progress in Clinical and Biological Research, 148. Alan R. Liss, New York, NY, pp. 523-537 ( 1984)), and the inhibitory effect of the test compound on the clinical score was evaluated according to the method of Koh et al. (Koh, CS, et al., Cell Immunology, 107 , 52-63 (1987)).
[0036]
The EAE animal model can cause EAE by sensitizing an animal with central nervous tissue (eg, guinea pig spinal cord) and adjuvant. First, symptoms occur with paralysis of the tail and lower limbs 9-10 days after immunization, followed by guinea-pig spinal cord (GPSC) homogenate and high-dose Mycobacterium tuberculosis (MT). ) Is used as a model for chronic progressive multiple sclerosis that repeats onset and recovery 2-3 times over about 40 days after immunization (Chabannes, D., et al. , J. Autoimmunity, 5, 199-211 ( 1992)).
[0037]
(1) Preparation of animals and experimental materials 1) A 0.5% -CMC solution (carboxymethyl cellulose solution) was used as a control. The solution was prepared by weighing 5.0 g of CMC (Serogen F-SB (Lot No. 154455), manufactured by Daiichi Kogyo Seiyaku Co., Ltd.) with an electronic balance (ER-182A; manufactured by Sartorius), and transferring the solution to a measuring cylinder. The solution was stirred and dissolved in distilled water (manufactured by Otsuka Pharmaceutical Factory Co., Ltd.) at 4 ° C. for a whole day and night to prepare a final concentration of 0.5%.
[0038]
2) Weigh the bulk of Compound 1 (Lot No. 5D96M), transfer it to an agate bowl, suspend it uniformly in a small amount of 0.5% CMC solution, and then add 0.5% CMC solution to the desired concentration. The final concentration of Compound 1 was adjusted to 60 mg / ml, dispensed into a sterile centrifuge tube, and stored in the dark at 4 ° C. until use.
[0039]
3) As the guinea pig spinal cord (GPSC) homogenate used as an immunogen, all 20 male Hartley guinea pigs (purchased from Nippon Steels River), 9-10 weeks old (500-600 g), were used. Guinea pigs were euthanized by cervical dislocation under light ether anesthesia, the meninges and blood vessels were detached from the spinal cord, and the excised spinal cord was transferred to an ice-cold petri dish. The amount of 10 spinal cords excised is weighed together, 1 ml of cold physiological saline (Otsuka Pharmaceutical Factory Co., Ltd.) is added to 1 g of GPSC in a potter type homogenizer, and a rotor for homogenate preparation (Ikemoto Rika Kogyo Co., Ltd.) is added. ) Was used to prepare a homogenate under ice cooling (GPSC concentration as 500 mg / ml). 3-4 ml each was dispensed into a PPP tube and stored frozen at -20 ° C.
[0040]
4) Preparation of tuberculosis-killed bacteria (MT) adjuvant was carried out using a commercially available tuberculosis-killed complete adjuvant solution (MT, H37Ra concentration: 1 mg / ml; catalog number 3113-60-5: manufactured by Difco) 10 ml / bottle, and tuberculosis 15 mg of killed bacteria H37Ra (Catalog No. 3114-33-8: manufactured by Difco) was added, and well-inverted after sealing with parafilm, and then suspended uniformly to prepare a final MT concentration of 2.5 mg / ml.
[0041]
5) Transfer the GPSC homogenate prepared in 3) and 4) and the tuberculosis-killed adjuvant solution to two glass syringes connected to a three-way stopcock at a ratio of 1: 1, and repeat until a uniform emulsion is formed. The emulsion solution obtained by mixing was used as an immune emulsion.
[0042]
6) A 10-week-old female Lewis rat (SPF: purchased from Charles River Japan) was used as an animal for the experiment. 100 animals were purchased at 9 weeks of age, and after acclimatization for 1 week, a total of 60 animals in each group of 20 animals / 4 cages X3 group were extracted for this study by stratified random extraction using the body weight as an index.
[0043]
(2) Preparation of an animal model for multiple sclerosis A female Lewis rat weighing 180-200 g was tested for 10 weeks of age. Rats were given free oral intake of water and normal diet.
[0044]
Experimental allergic encephalomyelitis (EAE) as an animal model of multiple sclerosis was prepared as follows. That is, the immunoemulsion prepared in the above (1) -5) is 0.2 ml in total with 0.1 ml each using a 23 gauge needle and a 1.0 ml glass syringe on the foot pads of rats with light ether anesthesia. As a 50 mg + MT, 250 μg / body was injected intradermally. After immunization, breeding was carried out at 5 animals / cage. The rats developed allergic cerebrospinal inflammation symptoms around 10 days after immunization, and repeated recurrence and remission of symptoms similar to human clinical symptoms, thus becoming an animal model of multiple sclerosis.
[0045]
(3) Examination of multiple sclerosis treatment effect and recurrence prevention effect of Compound 1 of the present invention 1) The tube is often overturned immediately before the suspension of Compound 1 of the present invention prepared in (1) -2) is administered to rats. After mixing, it was taken into a 1 ml or 2.5 ml Terumo syringe and 0.5 ml per 100 g of rat body weight was forcibly orally administered using an oral sonde for rats. The dose was determined for each animal according to the body weight of the day automatically input to the computer. A 0.5% CMC solvent administration group as a control group was also administered in the same manner.
[0046]
Experiments on an animal model of multiple sclerosis were carried out in a control group (CMC solvent group), CMC solvent was administered until 1-9 days after immunization, and 300 mg / kg / day of Compound 1 from the 10th day onset of EAE. CMC solvent was administered to the group administered immediately after the onset of the first symptom and the 1st to 16th day after immunization, and the compound 1 was observed from the 17th day when the symptom once recovered and then began to worsen again. In 3 groups of the group administered 300 mg / kg / day (group administered immediately before the onset of the second symptom), 20 mice per group were used orally once daily until 37 days after immunization. Was done.
[0047]
2) Body weight, blood cell measurement, thymus weight, spleen weight measurement and symptom observation (score determination)
From the test start date (immunization date) to the test end date (anatomy date), body weight and symptoms were observed (score determination) once a day. The body weight was measured with an electronic balance (Sartorius; LC-4200: manufactured by Sartorius) and automatically input to a connected personal computer (TOSHIBA J3100: manufactured by Toshiba). At the same time, the dose for each individual was determined and displayed.
[0048]
The symptom score was determined by observing the symptoms of rats in each group daily according to the evaluation criteria of Koh et al. (Koh, C.-S., et al., Cell Immunology, 107 , 52-63 (1987)). A score was determined for each of the rat tail and both hind limbs, and the total was recorded on the score recording sheet. Score determination is 0 for asymptomatic for the tail, 0.25 for relaxation to the middle, 0.5 for complete relaxation of the entire tail, 0 for asymptomatic, 1.0 for gait disturbance, Incomplete paralysis of both hind limbs and complete paralysis of one hind limb were evaluated as 2.0, and complete paralysis of both hind limbs was evaluated as 3.0. In addition, the death example in the test was excluded from the statistical processing of the score determination.
[0049]
3) Statistical processing method All statistical analysis was performed by SAS system (SAS Institute Japan, Ver.6.11).
[0050]
The body weight is the difference in the response over time by repeated analysis based on repeated measurements using the measured values for the first attack and subsequent compound 1 administration groups and the second attack and subsequent compound 1 administration groups, with the control group as the control. Was carried out at significance levels of 5% and 1%.
[0051]
Symptom observation (symptom score determination)
The entire study period was divided into two parts, the first half and the second half, according to the transition of the clinical score of the control group. That is, at the time of the most recovery between the two peaks on the score (Chabannes, D., et al., J. Autoimmunity, 5 , 199-211 (1992)) The analysis was performed in three periods, the first half and the second half. More specifically, the score determination results of each individual in each period are integrated, and an average integrated clinical score is obtained in each group (Branisteanu, DD, et al., J. Neuroimmunology, 61 , 151-160 (1995); Martin, D., et al., J. Neuroimmunology, 61 , 241-245 (1995)), one-way analysis of variance followed by two-sided Dunnett-test with control group as control ), And the significance level was 5% and 1%. The score was expressed as an average score and standard error.
[0052]
The blood count, thymus weight, and spleen weight were tested by a two-sided Dunnett test using the control group as a control after one-way analysis of variance, and the significance level was 5% and 1%.
[0053]
The symptom score determination result and the change in weight are shown in FIG. 1, FIG. 2, and FIG. 3, respectively.
[0054]
In FIG. 1, the vertical axis represents the mean clinical score, the horizontal axis represents the administration period (days) after immunization, (1) is the result of the control (CMC solvent only) group, and (2) is 300 mg / kg of Compound 1. (3) shows the results of the group in which administration of 300 mg / kg / day of Compound 1 was started on the 10th day after immunization, respectively.
[0055]
In FIG. 2, the vertical axis represents the average cumulative clinical score (average of clinical scores accumulated from 0 to 38 days), the horizontal axis represents (1): control (CMC solvent only) group, (2): 300 mg of Compound 1 A group in which administration of / kg / day was started from day 17 after immunization and a group in which administration of 300 mg / kg / day of Compound 1 was started from day 10 after immunization are shown.
[0056]
In FIG. 3, the vertical axis represents daily body weight change, the horizontal axis represents (1): control (CMC solvent only) group, (2) 300 mg / kg / day of Compound 1 was started on the 17th day after immunization. Groups and (3): groups in which administration of 300 mg / kg / day of Compound 1 was started from the 10th day after immunization are shown.
[0057]
From the above results, the following is clear. That is, as confirmed from FIG. 2, the compound 1 administration group showed a significant decrease in the average cumulative clinical score (P <0.01), that is, the symptom improving effect as compared with the control group. Further, as shown in FIG. 3, the change in body weight corresponds well with the change in the symptom score determination in FIG. 1, and the improvement tendency was recognized in the compound 1 administration group compared with the control group. Furthermore, as FIG. 1 shows, it turns out that the compound 1 has improved the symptom by administration immediately after the 1st symptom onset, and has suppressed the onset of the 2nd symptom. In addition, since the group administered with Compound 1 immediately before the onset of the second symptom suppresses recurrence of the symptom, the recurrence / recovery type that is difficult to treat clinically and the prevention of recurrence of chronic progressive multiple sclerosis and various It can be seen that a therapeutic effect on clinical symptoms can be expected.
[Brief description of the drawings]
1 is a graph showing the results of a test according to Pharmacological Test Example 1. FIG.
FIG. 2 is a graph showing the results of a test according to Pharmacological Test Example 1.
FIG. 3 is a graph showing the results of a test according to Pharmacological Test Example 1.

Claims (1)

General formula

[Wherein R represents a benzoyl group which may have a lower alkoxy group on the phenyl ring. The carbon-carbon bond at the 3-position and 4-position of the carbostyryl skeleton represents a single bond or a double bond. ]
A therapeutic agent for multiple sclerosis comprising a carbostyril derivative represented by the formula (1) and / or a salt thereof as an active ingredient.

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