JP4176170B2 - Medicine containing amino alcohol derivative and therapeutic agent for abnormal proliferative disease - Google Patents

Description

【００１８】
合成経路２
【化６】

【００１９】
合成経路３
【化７】

【００２０】
上記合成経路１の出発物質であるＮ−保護−２−アミノアルカンジオール類の合成法としては、Ｎ−保護−α−アミノケトン類を還元する方法（J.Org.Chem.,54,1866(1989)）、Ｎ−（ジフェニルメチレン）アミノ酸エステルを水素化ジイソブチルアルミニウム、続いてグリニャール試薬で処理する方法 （J.Org.Chem.,57,5469(1992)）、Ｎ−保護−アミノアルデヒドやＮ−保護−アミノ酸の酸塩化物に有機金属試薬を反応させる方法（J. Am. Chem. Soc., 95, 4098(1973)）、２−オキサゾリジノンとアルデヒドの不斉アルドール反応（J. Am. Chem. Soc., 108, 6757(1986)）（エバンス法）、キラルなイミダゾリジノンおよびオキサゾリジノンとアルデヒドの不斉アルドール反応（Helv. Chem. Acta, 70, 237(1987)）等が知られている。
一方、上記Ｎ−保護−α−アミノケトン類の合成法としては、Ｎ−保護−α−アミノ酸を出発原料としてアミノ酸のカルボキシル基を酸塩化物に変換後、ベンゼンとフリーデル−クラフツ反応させる方法（J.Am.Chem.Soc.,103,6157(1981)）、該アミノ酸のカルボキシル基をアルキルリチウム試薬で処理してリチウム塩とした後、グリニャール試薬と反応させる方法等が知られている（J.Org.Chem.,54,1866(1989)）。
【００２１】
一般式（Ｉ）で示される化合物の薬学的に許容される塩としては、塩酸、リン酸、硫酸、硝酸等の無機酸塩、ギ酸、酢酸、クエン酸、乳酸、リンゴ酸、シュウ酸、マレイン酸、フマル酸、コハク酸、トリフルオロ酢酸、メタンスルホン酸（メシル酸）、Ｐ−トルエンスルホン酸等の有機酸の塩をあげることができる。このような塩の製造は自体既知の方法によって行うことができ、例えば一般式（Ｉ）で示される化合物（遊離型）をアルコール等の適宜な溶媒に溶解し、通常等モル程度の上記の酸を添加して反応させ、所望により溶媒を溜去すればよい。
本発明化合物は遊離型である場合よりも、塩酸塩、クエン酸塩、乳酸塩、コハク酸塩等の塩型である場合の方が、水又は生理食塩水に対する溶解性が向上する。
【００２２】
〔作用〕
一般式（Ｉ）で示される本発明のアミノアルコール誘導体またはその薬学的に許容される塩（以下「本発明化合物」ということもある。）は、糖脂質の生合成を制御する作用を有し、該作用に基づく医薬としての有用性を有している。
一般式（Ｉ）で示される化合物は分子中に少なくとも２箇所に不斉炭素を有し、その立体配置（Ｌ−トレオ、Ｌ−エリトロ、Ｄ−トレオ、Ｄ−エリトロ）による各異性体は、それぞれ上記生合成制御作用が異なる。特に、本発明化合物のうち、糖脂質生合成阻害作用を有する化合物は、未分化な状態で異常増殖する細胞の分化を誘導する作用又は異常増殖細胞を正常化する作用を有し、異常増殖性疾患治療薬、特に癌治療薬として有用である。このような用途に好ましい本発明化合物は、その立体配置がＤ−トレオ又はＬ−トレオ体であり、Ｄ−トレオ体が特に好ましい。
なお、一般式（Ｉ）の化合物の立体配置、（１Ｓ、２Ｓ）、（１Ｒ、２Ｓ）、（１Ｓ、２Ｒ）又は（１Ｒ、２Ｒ）は、それぞれＬ−トレオ体、Ｌ−エリトロ体、Ｄ−エリトロ体又はＤ−トレオ体に相当する。
【００２３】
本発明化合物は未分化で異常増殖する細胞の分化を誘導する作用を有するので、本発明の治療薬は、ヒトを含む哺乳動物の異常増殖性疾患、すなわち細胞が未分化な状態で異常増殖を呈することに基づく各種疾患の処置、すなわちこのような疾患の治療、軽減（症状の改善）、維持（悪化防止）または予防を目的とする医薬品として使用される。このような疾患としては、良性腫瘍（子宮筋腫など）；固形癌（食堂癌、大腸癌、肺癌、胃癌、膵臓癌、肝癌等の扁平上皮癌や腺癌など、または脳腫瘍）、神経膠腫、白血病、悪性リンパ腫等の悪性腫瘍もしくは癌（癌腫、肉腫）；腎炎（糸球体腎炎など）、ヒト自己免疫性リンパ球増殖性症候群、リンパ球増殖性疾患、血管免疫芽細胞性リンパ節症、免疫芽細胞性リンパ節症、全身性エリテマトーデス、炎症性腸疾患（クローン病、潰瘍性大腸炎等）、進行性全身性硬化症、多発性筋炎（皮膚筋炎）、シェーグレン症候群、骨髄異形成症候群、強皮症、リウマチ、乾癬、創傷の過形成（肉芽等）などの異常増殖性疾患が例示されるが、本発明の治療薬は、好ましくは悪性腫瘍、特に固形癌の治療に有効である。
【００２４】
〔製剤化〕
本発明化合物を、担体、賦形剤、その他の添加物と共に、経口又は非経口的に投与する製剤とすることができる。
経口製剤としては、散剤、顆粒剤、カプセル剤、錠剤等の固形製剤；シロップ剤、エリキシル剤、乳剤等の液状製剤を挙げることができる。散剤は、例えば乳糖、デンプン、結晶セルロース、乳酸カルシウム、リン酸水素カルシウム、メタケイ酸アルミン酸マグネシウム、無水ケイ酸等の賦形剤と混合して得ることができる。顆粒剤は、上記賦形剤のほか、必要に応じて、例えば白糖、ヒドロキシプロピルセルロース、ポリビニルピロリドン等の結合剤や、カルボキシメチルセルロース、カルボキシメチルセルロースカルシウム等の崩壊剤を更に加え、湿式又は乾式で造粒して得ることができる。錠剤は、上記散剤又は顆粒剤をそのまま、又はステアリン酸マグネシウム、タルク等の滑沢剤を加えて打錠して得ることができる。また、上記錠剤又は顆粒剤は、ヒドロキシプロピルメチルセルロースフタレート、メタクリル酸メチルコポリマー、ヒドロキシプロピルメチルセルロースアセテート、ヒドロキシプロピルメチルセルロースサクシネート等の腸溶性基剤で被覆し、あるいはエチルセルロース、カルナウバロウ、硬化油、白色セラック等で被覆し、これらを腸溶性又は持続性製剤にすることができる。カプセル剤のうち硬カプセル剤は、上記散剤又は顆粒剤を硬カプセルに充填して得ることができ、また軟カプセル剤は、本発明化合物を、グリセリン、ポリエチレングリコール、ゴマ油、オリーブ油等に溶解し、これをゼラチン膜で被覆して得ることができる。
【００２５】
シロップ剤は、白糖、ソルビトール、グリセリン等の甘味剤と本発明化合物とを、水に溶解して得ることができる。また、甘味剤及び水のほかに、精油、エタノール等を加えてエリキシル剤とするか、あるいはアラビヤゴム、トラガント、ポリソルベート類（ポリソルベート２０、ポリソルベート６０、ポリソルベート８０（トウィーン８０）等）、カルボキシメチルセルロースナトリウム等を加えて乳剤又は懸濁剤にすることもできる。またこれらの液状製剤には必要に応じ、矯味剤、着色剤、保存剤等を加えることができる。
【００２６】
非経口製剤としては、注射剤、直腸投与剤、ペッサリー、皮膚外用剤、吸入剤、エアゾール剤、点眼剤等を挙げることができる。注射剤は、本発明化合物に、必要に応じてポリソルベート類等の非イオン界面活性剤；塩酸、水酸化ナトリウム、乳酸、乳酸ナトリウム、リン酸一水素ナトリウム、リン酸二水素ナトリウム等のpH調整剤；塩化ナトリウム、ブドウ糖等の等張化剤；アミノ酸類等の安定化剤；及び注射用蒸留水又は生理食塩水を加え、滅菌濾過した後、アンプルに充填して得ることができる。また更にマンニトール、デキストラン、ゼラチン等を加えて真空凍結乾燥し、用時溶解型の注射剤とすることができる。その他、粉末充填型の注射剤とすることもできる。また本発明化合物に、レシチン、ポリソルベート類、ポリオキシエチレン硬化ヒマシ油、マクロゴール等の乳化剤を加えた後、水中で乳化させた注射用乳剤にすることもできる。
【００２７】
また、注射剤としては、溶解性、目標臓器への移行速度の改善が可能なリポソーム製剤やリピッドマイクロスフェア等が挙げられる。リポソーム製剤は公知のリポソーム調製法（C.G. Knight, Liposomes: From Physical Structure to Therapeutic Applications, pp. 51-82, Elsevier, Amsterdam (1981); Proc. Natl. Acad. Sci., U.S.A., Vol.75, 4194(1978)）に従って調製することができる。
【００２８】
すなわち、リポソーム膜を形成する両親媒性物質としては、天然リン脂質（卵黄レシチン、大豆レシチン、スフィンゴミエリン、ホスファチジルセリン、ホスファチジルグリセロール、ホスファチジルイノシトール、ジホスファチジルグリセロール、ホスファチジルエタノールアミン、カルジオリピン等）、合成リン脂質（ジステアロイルホスファチジルコリン、ジパルミトイルホスファチジルコリン、ジパルミトイルホスファチジルエタノールアミン等）等のリン脂質が使用される。また、膜の安定性、流動性、薬剤の膜透過性を改善するために、コレステロール類（コレステロール、エルゴステロール、フィトステロール、シトステロール、スチグマステロール等）、リポソームに負電荷を付与することが知られている物質（ホスファチジン酸、ジセチルホスフェート等）、正電荷を付与することが知られている物質（ステアリルアミン、ステアリルアミンアセテート等）、酸化防止剤（トコフェロール等）、油性物質（大豆油、綿実油、ゴマ油、肝油等）等、公知の種々の添加剤を使用してもよい。
【００２９】
リポソームの製造は、例えば、以下の方法で行うことができる。上記両親媒性物質及び添加剤と、本発明化合物を、有機溶媒（クロロホルム、ジクロロメタン、エタノール、メタノール、ヘキサン等の単独又は混合溶媒）にそれぞれ溶解し、両溶液を混合し、フラスコ等の容器中において不活性ガス（窒素ガス、アルゴンガス等）の存在下で有機溶媒を除去し、器壁に薄膜を付着させる。次いで、この薄膜を適当な水性媒体（生理食塩水、緩衝液、リン酸緩衝生理食塩水等）に加え、撹拌機で撹拌する。小粒径のリポソームを得るためには、超音波乳化機、加圧型乳化機、フレンチプレス細胞破砕機等を用いて更に分散させる。このようにリポソーム化に必要な両親媒性物質等と本発明化合物が水性媒体に分散した液をメンブランフィルター処理することによってリポソーム化が進行し、粒径分布が制御されたナノスフェア−リポソーム（脂質超微粒子；粒径２５〜５０nm程度）を得ることができる。また、リポソームを限外濾過、遠心分離、ゲル濾過等の分画処理に付し、担持されなかった薬剤を除去してもよい。
【００３０】
また、膜形成物質として、上記両媒性物質、添加剤の他に、β−オクチルグルコシド、Ｌ−チロシン−７−アミド−４−メチルクマリン、フェニルアミノマンノシド又はスルファチドを添加することによって得られる、グルコース残基、チロシン残基、マンノース残基又はスルファチドを膜上に有するリポソームに本発明の一般式（Ｉ）のアミノアルコール誘導体を担持させることもできる（方法自体は、特開平４−６９３３２号参照）。
【００３１】
リピッドマイクロスフェアは、本発明化合物を大豆油、ゴマ油等に溶解し、天然リン脂質、グリセリン、水等を加え撹拌機で撹拌し、更に超音波乳化機、加圧型乳化機、フレンチプレス細胞破砕機等を用いて分散させることにより得られる。
【００３２】
直腸投与剤は、本発明化合物に、カカオ脂肪酸のモノ、ジ又はトリグリセリド、ポリエチレングリコール等の坐剤用基剤を加えた後、加温して溶融し、これを型に流し込んで冷却するか、あるいは本発明化合物を、ポリエチレングリコール、大豆油等に溶解した後、ゼラチン膜で被覆して得ることができる。
皮膚外用剤は、本発明化合物に、白色ワセリン、ミツロウ、流動パラフィン、ポリエチレングリコール等を加え、必要に応じ加温し、混練して得ることができる。
テープ剤は、本発明化合物に、ロジン、アクリル酸アルキルエステル重合体等の粘着剤を混練し、これを不織布等に展延して得ることができる。
吸入剤は、例えば薬学的に許容される不活性ガス等の噴射剤に、本発明化合物を溶解又は分散し、これを耐圧容器に充填して得ることができる。
【００３３】
〔投与方法〕
本発明化合物を有効成分として含む医薬の投与方法は、特に限定されないが、特に異常増殖性疾患の治療に使用する場合、筋肉内注射、静脈内注射、皮下注射又は腹腔内注射等の注射、経直腸投与、経肺投与などが好ましい。異常増殖性疾患の治療法としては、疾患部位に直接投与する方法（局注等）があげられる。
投与量は、患者の年令、健康状態、体重等に応じ適宜決定するが、一般には、０．２５〜２００mg/kg 、好ましくは０．５〜１００mg/kg を一日１回あるいはそれ以上に分けて投与する。
〔毒性〕
本発明医薬の有効成分である一般式（Ｉ）で示されるアミノアルコール誘導体は、薬理活性を示す投与量において、ほとんどもしくは全く細胞毒性を示さない。
【００３４】
【実施例】
次に本発明を実施例により更に詳細に説明するが、本発明はその要旨を越えない限り以下の実施例に限定されるものではない。
本発明化合物およびその合成中間体の製造法を調製例として示す。
以下の実施例においてＭｅＯＨ、ＡｃＯＥｔ、ＡｃＯＨ、ＤＭＦはそれぞれ、メタノール、酢酸エチル、酢酸、Ｎ，Ｎ−ジメチルホルムアミドを示す。
【００３５】
調製例１ （１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステルの合成
（１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール（２１．２ｇ，７０．３ｍｍｏｌ）をピリジン（３５０ｍｌ）に溶かし、氷溶上でメタンスルホニルクロリド（５．６ｍｌ，７２．３ｍｍｏｌ）を５分間かけて滴下した。氷浴上で３０分間攪拌した後、室温で一晩攪拌した。反応が終了していることをＴＬＣ（クロロホルム：メタノール＝２０：１）で確認した後、溶媒を留去し、酢酸エチル（５００ｍｌ）を加え、１Ｎ−ＨＣｌ（２５０ｍｌ×３）、飽和食塩水（２５０ｍｌ）で洗浄後、硫酸ナトリウム上で乾燥し、溶媒を留去した。析出した結晶を酢酸エチル：ヘキサン＝１：１で洗浄し、白色結晶の標記物質（２５．３ｇ，収率９５．０％）を得た。
TLC Rf 0.55(CHCl₃:MeOH=20:1)、 0.83(AcOEt), 0.62(Hexane:AcOEt=1:2)
¹H-NMR(CDCl₃)δ : 7.35-7.26(10H,m,aromatic), 5.30(1H,d,J=7.81Hz,NH), 5.02(2H,s,CH ₂-O-CO), 4.99(1H,d,J=3.91Hz,CH-OH), 4.43-4.39,4.22-4.12(3H,m,N-CH-CH ₂), 2.98(3H,s,SO₃CH₃)
なお、（１Ｓ、２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオールの代わりに、（１Ｒ、２Ｒ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオールを用いることにより、（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステルを合成することができた。
【００３６】
調製例２ （１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−モルホリノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（１．２１ｇ，３．１９ｍｍｏｌ）をＮ，Ｎ−ジメチルホルムアミド（６ｍｌ）に溶かし、室温下、モルホリン（１．１１ｇ，１２．８ｍｍｏｌ）を加え、４０℃で２４時間攪拌した。反応がほぼ終了していることをＴＬＣ（クロロホルム：メタノール＝２０：１，ヘキサン：酢酸エチル＝１：２，酢酸エチル）で確認した後、飽和炭酸水素ナトリウム溶液（７０ｍｌ）、酢酸エチル（１００ｍｌ）を加え、有機層を水、飽和食塩水で順次洗浄し、硫酸ナトリウム上で乾燥、ろ過した。溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー（ヘキサン：酢酸エチル＝１：２）で精製し、無色油状の標記物質（５０７．５ｍｇ，収率４３．０％）を得た。
TLC Rf 0.32(CHCl₃:MeOH=20:1)、 0.12(Hexane:AcOEt=1:2)
¹H-NMR(CDCl₃)δ : 7.38-7.26(10H,m,aromatic), 5.04(2H,s,CH₂O-CO), 5.00(1H,d,J=3.41Hz,H-1), 4.11(1H,m,H-2), 3.72(4H,m,(CH₂)₂O), 2.68-2.47(6H,m,(CH₂)₃N)
【００３７】
調製例３ （１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−ピロリジノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（１．５２ｇ，４．０１ｍｍｏｌ）をＤＭＦ（８ｍｌ）に溶かし、ピロリジン（１．１４ｇ，１６．０３ｍｍｏｌ）を加え、４０〜５０℃で１８時間攪拌した後、酢酸エチル（１００ｍｌ）を加え、有機層を飽和炭酸水素ナトリウム溶液（７０ｍｌ）、水（７０ｍｌ）、飽和食塩水（７０ｍｌ）で順次洗浄後、硫酸ナトリウム上で乾燥し、溶媒を減圧留去した。得られた粗生成物をシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝２０：１）で精製し、無色油状の標記物質（１．２１ｇ，収率８５．５％）を得た。
TLC Rf 0.20(CHCl₃:MeOH=20:1), 0.20(AcOEt)
¹H-NMR(CDCl₃)δ : 7.39-7.24(10H,m,aromatic), 5.06-5.02(2H,m,CH₂-O-CO), 4.99(1H,d,J=3.91Hz,H-1), 4.07(1H,m,H-2), 2.9-2.6(6H,m,(CH₂)₃N), 1.83-1.74(4H,m,H-3',H-4')
¹³C-NMR(CDCl₃)δ : 156.0，140.8，136.5，128.4，128.2，128.0，127.8，127.4，126.1，75.7，66.6，58.1，55.2，53.4，23.6
【００３８】
また、本化合物の塩酸塩は下記の方法で調製した。
（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−ピロリジノ−１−フェニル−１−プロパノール（９５．１ｍｇ、０．２６９ｍｍｏｌ）をエタノール（３ｍｌ）に溶かし、室温下、２Ｎ−塩酸（１６１．２μｌ、０．３２２ｍｍｏｌ）を加え、１０分間撹拌した後、溶媒を減圧留去した。この後エタノール（３ｍｌ）を加え、減圧留去する操作を３回繰り返し、室温下１６時間減圧乾燥し、淡黄色油状の（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−ピロリジノ−１−フェニル−１−プロパノール塩酸塩１０５．５ｍｇ（収率１００％）を得た。
【００３９】
調製例４ （１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−ピペリジノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（１．２１ｇ，３．１９ｍｍｏｌ）をＤＭＦ（６ｍｌ）に溶かし、ピペリジン（１．０９ｇ，１２．８ｍｍｏｌ）を加え、４０〜５０℃で２４時間攪拌した。反応がほぼ終了していることをＴＬＣ（酢酸エチル、クロロホルム：メタノール＝２０：１）で確認した後、酢酸エチル（１００ｍｌ）を加え、有機層を飽和炭酸水素ナトリウム溶液（７０ｍｌ）、水（７０ｍｌ）、飽和食塩水（７０ｍｌ）で順次洗浄後、硫酸ナトリウム上で乾燥し、溶媒を減圧留去した。得られた粗生成物をシリカゲルカラムクロマトグラフィー（酢酸エチル）で精製し、無色油状の標記物質（７９５．４ｍｇ，収率６８．０％）を得た。
TLC Rf 0.20(CHCl₃:MeOH=20:1), 0.17(AcOEt)
¹H-NMR(CDCl₃)δ : 7.36-7.25(10H,m,aromatic), 5.04(2H,s,CH₂-O-CO), 5.01(1H,d,J=3.42Hz,H-1), 4.94(1H,d,J=7.33Hz,NH), 4.15(1H,m,H-2), 2.64-2.45(6H,m,(CH₂)₃N), 1.68-1.54(4H,m,H-3',H-5'), 1.5-1.4(2H,m,H-4')
¹³C-NMR(CDCl₃)δ : 155.9，140.8，136.4，128.5，128.3，128.1，127.9，127.4，126.3，75.7，66.7，60.5，55.8，51.7，26.1，23.9
【００４０】
調製例５ （１Ｓ，２Ｓ）−２−ｔ−ブトキシカルボニルアミノ−３−モルホリノ−１−フェニル−１−プロパノールの合成
調製例１の方法に従い（１Ｓ，２Ｓ）−２−ｔ−ブトキシカルボニルアミノ−１−フェニル−１，３−プロパンジオールをメシル化した後、調製例２の方法に従ってモルホリン置換反応を行った結果、無色油状の標記物質を収率６３％で得た。
TLC Rf 0.36(CHCl₃:MeOH=20:1)
¹H-NMR(CDCl₃)δ : 7.38-7.26(5H,m,aromatic), 4.98(1H,d,J=3.91Hz,H-1), 4.05(1H,m,H-2), 3.74(4H,m,(CH₂)₂O), 2.64-2.59(5H,m,H-2',H-6',H-3A), 2.46(1H,dd,J=4.89,13.19Hz,H-3B), 1.38(9H,s,(CH₃)₃)
【００４１】
調製例６ （１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−３−（Ｎ−メチルピペラジノ）−１−フェニル−１−プロパノールの合成
（１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（１．８１ｇ，４．７８ｍｍｏｌ）をエタノール（４０ｍｌ）に溶かし、ヨウ化ナトリウム（７１２．８ｍｇ，４．７５ｍｍｏｌ）、Ｎ−メチルピペラジン（１．９２ｇ，１９．２ｍｍｏｌ）を加え、５０℃で５日間攪拌した。反応状況をＴＬＣ（クロロホルム：メタノール＝９：１）で確認した後、溶媒を留去し、水（５０ｍｌ）、酢酸エチル（１００ｍｌ）を加え、有機層を水、飽和食塩水で順次洗浄し、硫酸ナトリウム上で乾燥、ろ過した。溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝２０：１）で精製し、無色油状の標記物質（２４２．２ｍｇ，収率１３．２％）を得た。
TLC Rf 0.38(CHCl₃:MeOH=9:1)
¹H-NMR(CDCl₃)δ : 7.36-7.26(10H,m,aromatic), 5.04(2H,s,CH₂-O-CO), 5.00(1H,d,J=3.41Hz,H-1), 4.97(1H,d,NH), 4.12(1H,m,H-2), 2.70-2.49(10H,m,(CH₂)₃N,(CH₂)₂N), 2.28(3H,s,CH₃-N)
¹³C-NMR(CDCl₃)δ : 156.0，140.7，136.4，128.5，128.3，128.1，127.9，127.5，126.2，75.3，66.8，59.6，55.1，54.1，52.1，45.9
【００４２】
調製例７ （１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−３−（（２Ｓ）−２−ヒドロキシメチルピロリジノ）−１−フェニル−１−プロパノールの合成
（２Ｓ）−２−ヒドロキシメチルピロリジン（３２３．３ｍｇ，３．２０ｍｍｏｌ）をエタノール（１２ｍｌ）に溶かし、（１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−ｐ−ブロモベンゼンスルホニルエステル（７８２ｍｇ，１．５０ｍｍｏｌ）の塩化メチレン溶液（３ｍｌ）に滴下した。４５℃で２日間攪拌し、反応状況をＴＬＣ（クロロホルム：メタノール＝９：１、酢酸エチル：２−プロパノール＝２：１）で確認した後、溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル：２−プロパノール＝７：３）で精製し、無色油状の標記物質（７９．５ｍｇ，収率１３．８％）を得た。
TLC Rf 0.25(CHCl₃:MeOH=9:1), 0.39(AcOEt:(CH₃)₂CHOH=2:1)
¹H-NMR(CDCl₃)δ : 7.51-7.23(10H,m,aromatic), 5.32(1H,br,NH), 4.99(3H,m,H-1,CH₂-O-CO), 3.93(1H,m,H-2), 3.67,3.66,3.64,3.63(1H,dd,CH ₂(A)-OH), 3.51(1H,dd,J=4.40,11.23Hz,CH ₂(B)-OH), 3.28-3.23(1H,m,H-5'A), 3.08(1H,dd,J=5.86,13.19Hz,H-3A), 2.81(1H,dd,J=2.93,13.18Hz,H-3B), 2.71(1H,m,H-2'), 2.34-2.28(1H,m,H-5'B), 1.90-1.59(4H,m,H-3',H-4')
¹³C-NMR(CDCl₃)δ : 156.5，141.0，136.5，128.4，128.3，128.0，127.8，127.5，125.8，75.4，66.6，66.4，63.7，58.0，56.2，55.4，27.0，23.8
【００４３】
調製例８ （１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−３−（３−ヒドロキシピロリジノ）−１−フェニル−１−プロパノールの合成
（１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（２．６０ｇ，６．８６ｍｍｏｌ）をエタノール（２０ｍｌ）に溶かし、３−ヒドロキシピロリジン（１．１９ｇ，１３．６８ｍｍｏｌ）を加え、４５℃で５日間攪拌した。反応状況をＴＬＣ（クロロホルム：メタノール＝９：１）で確認した後、溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー（クロルホルム：メタノール＝９：１、酢酸エチル：メタノール＝９：１）で精製し、無色油状の標記物質（５２７．１ｍｇ，収率２０．８％）を得た。
TLC Rf 0.25(CHCl₃:MeOH=9:1), 0.35(AcOEt:MeOH=4:1)
¹H-NMR(CDCl₃)δ : 7.41-7.24(10H,m,aromatic), 5.26(0.7H,d,J=7.82Hz,NH,片方のジアステレオマー由来), 5.20(0.3H,d,NH,もう片方のジアステレオマー由来), 5.00(3H,s,H-1,CH₂-O-CO), 4.34(0.7H,m,H-3',片方のジアステレオマー由来), 4.28(0.3H,m,H-3',もう片方のジアステレオマー由来), 4.02(1H,m,H-2), 3.04-2.99, 2.89-2.42(6H,m,(CH₂)₃N), 2.20-2.07(1H,m,H-4'A), 1.80-1.68(1H,m,H-4'B)
¹³C-NMR(CDCl₃)δ : 156.5，141.2，141.1，136.7，128.8，128.6，128.3，128.2，127.8，126.4，75.4，75.2，71.3，67.0，64.0，58.0，54.2，54.1，53.8，34.9
【００４４】
調製例９ （１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−３−ピロリジノ−１−フェニル−１−プロパノールの合成
（１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（１．２１ｇ，３．１９ｍｍｏｌ）をＤＭＦ（６ｍｌ）に溶かし、ピロリジン（０．９１ｇ，１２．８ｍｍｏｌ）を加え、４０℃で２４時間攪拌した後、酢酸エチル（１００ｍｌ）を加え、有機層を飽和炭酸水素ナトリウム溶液（７０ｍｌ）、水（７０ｍｌ）、飽和食塩水（７０ｍｌ）で順次洗浄後、硫酸ナトリウム上で乾燥し、溶媒を減圧留去した。得られた粗生成物をシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝２０：１）で精製し、無色油状の標記物質（９８３．１ｍｇ，収率８７．０％）を得た。
TLC Rf 0.20(CHCl₃:MeOH=20:1), 0.20(AcOEt)
¹H-NMR(CDCl₃)δ : 7.39-7.24(10H,m,aromatic), 5.06-5.02(2H,m,CH₂-O-CO), 4.99(1H,d,J=3.91Hz,H-1), 4.07(1H,m,H-2), 2.9-2.6(6H,m,(CH₂)₃N), 1.83-1.74(4H,m,H-3',H-4')
¹³C-NMR(CDCl₃)δ : 156.0，140.8，136.5，128.4，128.2，128.0，127.8，127.4，126.1，75.7，66.6，58.1，55.2，53.4，23.6
【００４５】
調製例１０ （１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−３−（３−ヒドロキシメチルピペリジノ）−１−フェニル−１−プロパノールの合成
（１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（２．４３ｇ，６．４１ｍｍｏｌ）をエタノール（２０ｍｌ）に溶かし、３−ヒドロキシメチルピペリジン（１．４７ｇ，１２．７８ｍｍｏｌ）を加え、４５℃で５日間攪拌した。反応状況をＴＬＣ（クロロホルム：メタノール＝９：１）で確認した後、溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー（クロルホルム：メタノール＝２０：１、酢酸エチル：メタノール＝２０：１）で精製し、無色油状の標記物質（２９３．３ｍｇ，１１．５％）を得た。
TLC Rf 0.42(CHCl₃:MeOH=9:1), 0.16(AcOEt:MeOH=20:1)
¹H-NMR(CDCl₃)δ : 7.35-7.26(10H,m,aromatic), 5.03(2H,s,CH₂-O-CO), 4.994(0.5H,d,J=7.81Hz,H-1,片方のジアステレオマー由来), 4.986(0.5H,d,J=8.30Hz,H-1,もう片方のジアステレオマー由来), 4.15-4.09(1H,m,H-2), 3.56-3.45(2H,m,CH ₂-OH), 3.00-2.91, 2.75, 2.25-2.00(4H,m,H-2',H-6'), 2.65-2.59(1H,m,H-3A), 2.49-2.45(1H,m,H-3B), 1.82(1H,m,H-3'), 1.75-1.65, 1.63-1.53, 1.09-1.04(4H,m,H-4',H-5')
¹³C-NMR(CDCl₃)δ : 156.2，156.1，140.7，136.4，128.5，128.3，128.1，127.9，127.5，126.2，75.4，75.3，66.7，65.7，65.6，60.4，60.3，60.2，58.2，57.5，55.7，55.1，52.0，38.8，38.7，26.6，24.7，14.2
【００４６】
調製例１１ （１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−３−（４−ヒドロキシピペリジノ）−１−フェニル−１−プロパノールの合成
（１Ｓ，２Ｓ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（２７３．６ｍｇ，０．７２２ｍｍｏｌ）をエタノール（３ｍｌ）に溶かし、ヨウ化ナトリウム（１１９．２ｍｇ，０．７９５ｍｍｏｌ）、４−ヒドロキシピペリジン（１７１．５ｍｇ，１．７０ｍｍｏｌ）を加え、室温で４日間攪拌した。反応状況をＴＬＣ（クロロホルム：メタノール＝９：１）で確認した後、４−ヒドロキシピペリジン（１５７．０ｍｇ，１．５５ｍｍｏｌ）を追加し、４５℃でさらに２日間攪拌した。溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝２０：１）で精製し、無色油状の標記物質（１１２．６ｍｇ，収率４０．６％）を得た。
TLC Rf 0.24(CHCl₃:MeOH=9:1)
¹H-NMR(CDCl₃)δ : 7.36-7.25(10H,m,aromatic), 5.03(3H,m,CH₂-O-CO,NH), 5.00(1H,d,J=2.93Hz,H-1), 4.11(1H,m,H-2), 3.71(1H,m,H-4'), 2.91, 2.82，2.64，2.48，2.32(6H,m,(CH₂)₃N), 1.89(2H,m,H-3'A,H-5'A), 1.64-1.56(2H,m,H-3'B,H-5'B)
¹³C-NMR(CDCl₃)δ : 156.1，140.7，136.4，128.5，128.3，128.1，127.9，127.5，126.2，75.4，66.9，66.8，66.7，59.5，52.2，51.9，34.4
【００４７】
調製例１２ （１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−（４−ヒドロキシピペリジノ）−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−１，３−プロパンジオール−３−メタンスルホニルエステル（２９５．６ｍｇ，０．７８０ｍｍｏｌ）をエタノール（３ｍｌ）に溶かし、４−ヒドロキシピペリジン（３１５．１ｍｇ，３．１２ｍｍｏｌ）を加え、４５℃で２日間攪拌した。反応状況をＴＬＣ（クロロホルム：メタノール＝９：１）で確認した後、溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝２０：１）で精製し、無色油状の標記物質（１７９．７ｍｇ，収率６０．０％）を得た。
TLC Rf 0.24(CHCl₃:MeOH=9:1)
¹H-NMR(CDCl₃)δ : 7.36-7.25(10H,m,aromatic), 5.03(3H,m,CH₂-O-CO,NH), 5.00(1H,d,J=2.93Hz,H-1), 4.11(1H,m,H-2), 3.71(1H,m,H-4'), 2.91, 2.82，2.64，2.48，2.32(6H,m,(CH₂)₃N), 1.89(2H,m,H-3'A,H-5'A), 1.64-1.56(2H,m,H-3'B,H-5'B)
¹³C-NMR(CDCl₃)δ : 156.1，140.7，136.4，128.5，128.3，128.1，127.9，127.5，126.2，75.4，66.9，66.8，66.7，59.5，52.2，51.9，34.4
【００４８】
調製例１３ （１Ｒ，２Ｒ）−２−アミノ−３−ピロリジノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−ピロリジノ−１−フェニル−１−プロパノール（３９４．２ｍｇ，１．１１ｍｍｏｌ）をメタノール（１０ｍｌ）に溶かし、１０％パラジウム炭素（１２４．８ｍｇ，１０．６ｍｏｌ％）を加え、水素雰囲気下、室温で30分間撹拌した。TLC（クロロホルム：メタノール＝７：３および酢酸エチル：メタノール＝２：１）で反応が終了していることを確認した後、パラジウム炭素をろ過除去した。ろ液を濃縮し、無色油状の標記物質（２５０．２ｍｇ）を得た。
【００４９】
調製例１４ （１Ｒ，２Ｒ）−２−オクチルオキシカルボニルアミノ−３−ピロリジノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−アミノ−３−ピロリジノ−１−フェニル−１−プロパノール（２５０．２ｍｇ，１．１１ｍｍｏｌ）をメタノール（５ｍｌ）に溶かし、室温下、トリエチルアミン（０．１８６ｍｌ，１．３３７ｍｍｏｌ）を加えた後、氷浴上にてクロロぎ酸ｎ−オクチルエステル（０．２４０ｍｌ，１．２２６ｍｍｏｌ）を加え、室温下攪拌した。９０分後、トリエチルアミン（０．１８６ｍｌ，１．３３７ｍｍｏｌ）、クロロぎ酸ｎ−オクチルエステル（０．２４０ｍｌ，１．２２６ｍｍｏｌ）追加し、室温下攪拌した。２０時間後、溶媒を減圧留去し、酢酸エチル（１００ｍｌ）を加え、有機層を飽和炭酸水素ナトリウム溶液、水、飽和食塩水それぞれ７０ｍｌで順次洗浄し、硫酸ナトリウム上で乾燥後、ろ過し、溶媒を減圧留去した。得られた粗生成物をシリカゲルカラムクロマトグラフィーで精製し（溶出溶媒；クロロホルム：メタノール＝２０：１）、無色油状の標記物質（９９．５ｍｇ、収率２３．８％）を得た。
TLC Rf 0.28(AcOEt:MeOH=4:1)、0.30(CHCl₃:MeOH=9:1)
¹H-NMR(CDCl₃)δ:7.36-7.24(5H,m,aromatic)，5.05(1H,d,J=2.93Hz,H-1)，4.9(1H,d,NH)，4.04(1H,m,H-2)，3.96(2H,m,COOCH₂)，2.91-2.68(6H,m,CH₂N(CH₂)₂)，
1.80(4H,m,H-3',H-4')，1.52(2H,m,COOCH₂CH ₂)，1.26(10H,m,(CH ₂)₅CH₃)，0.88(3H,t,CH₂CH ₃)
¹³C-NMR(CDCl₃)δ : 156.4，140.9，128.2，127.3，126.1，75.6，65.1，58.1，55.2，53.3，31.7，29.1，28.9，25.7，23.6，22.6，14.0
【００５０】
調製例１５ （１Ｒ，２Ｒ）−２−デシルアミノ−３−ピロリジノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−デカノイルアミノ−３−ピロリジノ−１−フェニル−１−プロパノール（１８１．８ｍｇ，０．４８６ｍｍｏｌ）を塩化メチレン（５ｍｌ）に溶かし、室温下、水素化リチウムアルミニウム（１５３．０ｍｇ，４．０３２ｍｍｏｌ）を加え、３５〜４０℃で２．５時間還流後、氷浴上にて１Ｎ−塩酸（１５ｍｌ）を加え、３０分間攪拌した後、飽和炭酸水素ナトリウム溶液（７０ｍｌ）、クロロホルム（１００ｍｌ）を加え、有機層を水、飽和食塩水それぞれ７０ｍｌで順次洗浄し、硫酸ナトリウム上で乾燥後、ろ過し、溶媒を減圧留去した。得られた粗生成物をシリカゲルカラムクロマトグラフィーで精製し（溶出溶媒；クロロホルム：メタノール＝２０：１、酢酸エチル：メタノール＝２：１）、無色油状の標記物質（１２１．２ｍｇ、収率６９．３％）を得た。
TLC Rf 0.39(CHCl₃:MeOH=9:1)、0.19(AcOEt:MeOH=2:1)
¹H-NMR(CDCl₃)δ:7.37-7.22(5H,m,aromatic)，4.68(1H,d,J=3.90Hz,H-1)，2.99(1H,m,H-2)，2.63-2.42(8H,m,CH₂N(CH₂)₂,NHCH ₂)，1.77(4H,m,H-3',H-4')，1.41-1.24（16H,m,(CH ₂)₈CH₃），0.88(3H,t,CH₃)
¹³C-NMR(CDCl₃)δ : 143.1，128.1，127.0，126.2，73.9，61.2，57.6，54.5，48.5，31.9，30.2，29.7，29.6，29.4，29.3，27.1，23.6，22.7，14.1
【００５１】
調製例１６ （１Ｒ，２Ｒ）−２−ヘキサノイルアミノ−３−ピロリジノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−アミノ−３−ピロリジノ−１−フェニル−１−プロパノール（１６０．０ｍｇ，０．７２７ｍｍｏｌ）をメタノール（４ｍｌ）に溶かし、室温下、トリエチルアミン（０．１２１ｍｌ，０．８７３ｍｍｏｌ）を加えた後、氷浴上にてｎ−ヘキサノイルクロリド（０．１０２ｍｌ，０．７２７ｍｍｏｌ）を加え、室温下攪拌した。２時間後、トリエチルアミン（０．１２１ｍｌ，０．８７３ｍｍｏｌ）、ｎ−ヘキサノイルクロリド（０．０５ｍｌ，０．３５８ｍｍｏｌ）を追加し、室温下攪拌した。９０分後後、メタノール（５ｍｌ）を加え、溶媒を減圧留去し、酢酸エチル（４０ｍｌ）を加え、有機層を飽和炭酸水素ナトリウム溶液、水、飽和食塩水それぞれ２５ｍｌで順次洗浄し、硫酸ナトリウム上で乾燥後、ろ過し、溶媒を減圧留去した。得られた粗生成物をシリカゲルカラムクロマトグラフィーで精製し（溶出溶媒；酢酸エチル：メタノール＝２：１）、無色油状の標記物質（２１．７ｍｇ、収率９．４％）を得た。
TLC Rf 0.12(CHCl₃:MeOH=9:1)、0.23(AcOEt:MeOH=2:1)
¹H-NMR(CDCl₃)δ:7.36-7.23(5H,m,aromatic)，5.86(1H,d,J=7.33Hz,NH),5.04(1H,d,J=2.93Hz,H-1)，4.27-4.22(1H,m,H-2)，2.85-2.83(2H,m)，2.71-2.63(4H,m),2.09-2.04(2H,m,COCH₂)，1.79(4H,m),1.52-1.44(2H,m,COCH₂CH ₂),1.29-1.20(2H,m),1.17-1.11(2H,m),0.84(3H,t,CH₃)
¹³C-NMR(CDCl₃)δ : 173.4, 141.1, 128.2, 127.4, 125.9, 75.8, 58.1, 55.3, 52.4, 36.7, 31.2, 25.3, 23.7, 22.3, 13.8
【００５２】
調製例１７ （１Ｒ，２Ｒ，２’Ｓ）−２−（２’−ヒドロキシオクタノイルアミノ）−３−ピロリジノ−１−フェニル−１−プロパノールおよび（１Ｒ，２Ｒ，２’Ｒ）−２−（２’−ヒドロキシオクタノイルアミノ）−３−ピロリジノ−１−フェニル−１−プロパノールの合成
（１Ｒ，２Ｒ）−２−アミノ−３−ピロリジノ−１−フェニル−１−プロパノール（１７９．０ｍｇ，０．８１４ｍｍｏｌ）を塩化メチレン（６ｍｌ）に溶かし、室温下、２−ヒドロキシオクタン酸（１６６．３ｍｇ，１．０８３ｍｍｏｌ）、Ｎ−ヒドロキシスクシンイミド（１９８．２ｍｇ，１．７２２ｍｍｏｌ）を加え２０分間撹拌した後、氷浴上で１−エチル−３−（３−ジメチルアミノプロピル）カルボジイミド塩酸塩（１７１．１ｍｇ，０．８９３ｍｍｏｌ）を加え、室温下一夜攪拌した。ＴＬＣ（クロロホルム：メタノール＝７：３）で反応がほとんど終了していることを確認した後、飽和炭酸水素ナトリウム溶液（２０ｍｌ）を加え、クロロホルムで抽出（３０ｍｌｘ３）した。有機層を硫酸ナトリウム上で乾燥後、ろ過し、溶媒を減圧留去した。得られた粗生成物をシリカゲルカラムクロマトグラフィーで精製し（溶出溶媒；クロロホルム：メタノール＝２０：１、酢酸エチル：メタノール＝２：１）、無色油状の標記物質（（１Ｒ，２Ｒ，２’Ｓ）体を６．９ｍｇ、（１Ｒ，２Ｒ，２’Ｒ）体を８．８ｍｇ）を得た。
（１Ｒ，２Ｒ，２’Ｓ）−２−（２’−ヒドロキシオクタノイルアミノ）−３−ピロリジノ−１−フェニル−１−プロパノール
TLC Rf 0.22(CHCl₃:MeOH=9:1)、0.32(AcOEt:MeOH=2:1)
¹H-NMR(CDCl₃)δ:7.34-7.23(5H,m,aromatic)，6.72(1H,d,NH),5.08(1H,d,J=2.92Hz,H-1)，4.24(1H,m,H-2)，3.95(1H,m,H-2'), 2.98-2.85(2H,m)，2.73-2.65(4H,m), 1.80(4H,m),1.64(1H,m,H-3'A),1.42(1H,m,H-3'B),1.28-1.21(8H,m,(CH ₂)₄CH₃),0.87(3H,t,CH₃)
¹³C-NMR(CDCl₃)δ : 174.1, 141.0, 128.3, 127.4, 125.8, 75.6, 71.9, 58.2, 55.4, 52.5, 34.8, 31.6, 29.0, 24.7, 23.7, 22.6, 14.0
【００５３】
調製例１８ （１Ｒ，２Ｒ，２’Ｒ）−２−（２’−ヒドロキシオクタノイルアミノ）−３−ピロリジノ−１−フェニル−１−プロパノールの合成
調製例１７の方法に従い、２−ヒドロキシオクタン酸の代わりに、（２Ｒ）−２−ヒドロキシオクタン酸を用い、標記化合物を合成すると同時に、調製例１７で合成、分離した化合物を同定した。
（１Ｒ，２Ｒ，２’Ｒ）−２−（２’−ヒドロキシオクタノイルアミノ）−３−ピロリジノ−１−フェニル−１−プロパノール
TLC Rf 0.32(CHCl₃:MeOH=4:1)、0.20(AcOEt:MeOH=2:1)
¹H-NMR(CDCl₃)δ:7.33-7.23(5H,m,aromatic)，6.97(1H,d,NH),5.02(1H,d,J=3.41Hz,H-1)，4.35(1H,m,H-2)，4.03(1H,m,H-2'), 3.09-3.04(1H,m),2.87-2.78(5H,m)，1.85(4H,brs),1.58(1H,m,H-3'A),1.44(1H,m,H-3'B),1.27-1.21(8H,m,(CH ₂)₄CH₃),0.87(3H,t,CH₃)
¹³C-NMR(CDCl₃)δ : 174.4, 140.6, 128.3, 127.6, 125.9, 75.1, 72.3, 57.9, 55.2, 51.7, 34.4, 31.6, 29.0, 24.6, 23.5, 22.6, 14.0
【００５４】
調製例１９ （１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−３−ピロリジノ−１−フェニル−１−プロピル アセテートの合成
（１Ｒ，２Ｒ）−２−ベンジルオキシカルボニルアミノ−１−フェニル−３−ピロリジノ−１−プロパノール２８４．２ｍｇ（０．８０３ｍｍｏｌ）をピリジン５ｍｌに溶かし、氷浴上にて、無水酢酸１５１．４μｌ（１．６１ｍｍｏｌ）を加え、室温下撹拌し、１６時間後、無水酢酸３８．０μｌ（０．４０３ｍｍｏｌ）を追加した。反応終了後、１Ｎ 塩酸３０ｍｌを加え、２０分間撹拌した後、酢酸エチル５０ｍｌで抽出し、有機層を水、飽和炭酸水素ナトリウム溶液、水、飽和食塩水それぞれ３０ｍｌで順次洗浄し、硫酸ナトリウム上で乾燥後、ろ過し、溶媒を減圧留去した。また、合わせた洗液をクロロホルム７０ｍｌで３回抽出し、有機層を硫酸ナトリウム上で乾燥後、ろ過し、溶媒を減圧留去した。得られた粗生成物を合わせてシリカゲルカラムクロマトグラフィー（クロロホルム：メタノール＝２０：１、酢酸エチル：メタノール＝２０：１）で精製し、無色油状の標記物質２３０．９ｍｇ（収率７２．６％）を得た。
TLC Rf. 0.24（CHCl₃：MeOH＝20:1）、0.31（AcOEt：MeOH＝9:1)
¹H-NMR(CDCl₃):7.34-7.25(10H,m,aromatic)，5.95(1H,d,J=4.88Hz,H-1)，5.10-4.94(2H,m,COOCH₂)，4.17(1H,m,H-2)，2.51(6H,m,CH₂N(CH₂)₂),2.03(3H,s,COCH₃),1.73(4H,m,H-3',H-4')
¹³C-NMR(CDCl₃): 169.7, 156.1，137.7, 136.6，128.6, 128.3，128.2, 128.0，127.9, 126.6，75.3，66.5，56.5，54.5，54.3，23.5，20.8
【００５５】
実施例１
本発明化合物とＤ−トレオ−ＰＤＭＰのグルコシルセラミド合成酵素に対する阻害効果及び細胞毒性を比較検討した。
〔方法及び結果〕
グルコシルセラミド合成酵素に対する阻害活性は、マウス肝臓のミクロゾーム画分を酵素源とし、井ノ口とRadinによって報告された方法（J. Lipid Res. vol.28, 565-571, 1987）によって測定し、各化合物のグルコシルセラミド合成酵素に対する５０％酵素活性阻害濃度（ＩＣ₅₀）として求めた。また、各化合物の培養ヒト胎児繊維芽細胞に対する細胞毒性を２４時間培養後、ＭＴＴ法（Pauwels et al, J. Virol. Methods, Vol. 20, pp.309-321,(1988)）により、５０％の細胞が死滅する濃度（ＣＤ₅₀）として求めた。さらに、細胞毒性を示す濃度が高く、グルコシルセラミド合成酵素を阻害する濃度が低ければ、その化合物の安全性が高いと言えることから、ＣＤ₅₀／ＩＣ₅₀値を算出した。これらの結果を表１に示した。その結果、本発明の化合物は、Ｄ−トレオ−ＰＤＭＰに比較して、グルコシルセラミド合成酵素に対する阻害活性が著しく高く、さらに細胞毒性は調べた濃度範囲では全く認められなかった。ＣＤ₅₀／ＩＣ₅₀値においてもＤ−トレオ−ＰＤＭＰは５であるのに対し、本発明の化合物はいずれも高く、安全性においても優れていることが判明した。
【００５６】
【表１】

【００５７】
実施例２
調製例３の本発明化合物のヒト食道癌に対する治療効果を検討した。
〔方法及び結果〕
ヒト食道癌より樹立された細胞株ＫＥＳ−３を１ｘ１０⁷個をヌードマウスの側背部皮下に移入し、１〜２週間後の腫瘍形成時に調製例３の化合物の塩酸塩の生理食塩水溶液（７．７８ｍｇ／ｍｌ）１０μｌを１日１回、７日間連続局注し、病理組織学的変化を検討した。比較対照としては、溶媒の生理食塩水を同量局注した。その結果を光学顕微鏡写真１及び２（倍率２４０）により示す。
ＫＥＳ−３の上記投与実験の移植組織像（ヘマトキシエオジン染色）を比較してみると、生理食塩水対照群の組織像（写真１）と比べて調製例３の本発明化合物投与群は全般的に核の空ほう化や凝縮、および細胞膨化を呈しており、明らかな変性が顕著に認められた（写真２）。
【００５８】
【発明の効果】
本発明によれば一般式（Ｉ）で示されるアミノアルコール誘導体またはその薬学的に許容される塩を有効成分として含む医薬、特に悪性腫瘍のように未分化な状態で異常な細胞増殖を呈することに基づく疾患の治療に有効な医薬、さらには固形癌治療薬として有望な医薬を提供することができる。
【図面の簡単な説明】
【図１】ＫＥＳ−３をヌードマウスに移植後、生理食塩水を局注した場合の病理組織学的な変化を表す光学顕微鏡写真１である。
【図２】ＫＥＳ−３をヌードマウスに移植後、調製例３の化合物の塩酸塩の生理食塩水溶液を局注した場合の病理組織学的な変化を表す光学顕微鏡写真２である。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a medicament containing an amino alcohol derivative or a pharmaceutically acceptable salt thereof and a therapeutic agent for an abnormal proliferative disease. Specifically, the present invention induces differentiation of a glycosphingoglycolipid (hereinafter, also referred to as “glycolipid”) biosynthesis-controlling derivative, particularly a cell that proliferates abnormally in an undifferentiated state. Furthermore, the present invention relates to a therapeutic agent for an abnormal proliferative disease, which is intended to differentiate from terminally differentiated cells having a certain life span and to escape from an abnormally proliferative state, and further to a therapeutic agent for cancer.
[0002]
[Prior art]
Glucosphingoglycolipid is present as a cell surface membrane constituent of mammalian cells, and is generated, proliferated, differentiated through the receptor function of biologically active substances, the mutual recognition function between cells, or the interaction between cells, etc. It is known to be closely related to cell functions such as canceration and immune reaction.
Currently, the most commonly used method for exploring the function of glycolipids is to add glycolipids from the outside to the experimental system. In this case, the relationship with endogenous glycolipids becomes a problem. . In other words, while the endogenous glycolipid present in the cell membrane has already formed a complex with various cell surface receptors, etc., the result derived by adding additional glycolipid is the true result of endogenous glycolipid. It may not always reflect the cellular physiological significance of. Therefore, in order to know the original role of glycolipids in cell physiology, a method for specifically inhibiting endogenous glycolipid biosynthesis was required.
[0003]
The present inventors first synthesized various 2-acylaminopropanol derivatives, which are ceramide analogs, and examined their glycolipid biosynthesis inhibitory activity, thereby producing 1-phenyl-2-decanoylamino-3-morpholino. Among the stereoisomers of -1-propanol (PDMP), D- or DL-threo form specifically inhibits glucosylceramide biosynthetic enzymes, and the intracellular content of all glycolipids starting from glucosylceramide (Adv. Lipid Res., 26, 183-213, 1993). The use of PDMP or an analog thereof as a cancer therapeutic agent is disclosed in JP-A-1-254623. Furthermore, PDMP has the effect of increasing the intracellular content of ceramide, which is a biosynthetic precursor of glucosylceramide, by inhibiting glucosylceramide synthesis, but in recent years ceramide molecules are involved in the suppression of differentiation, apoptosis and cell proliferation. Many evidences as important intracellular signal transduction molecules in the intracellular signal transduction system have been reported (Immunology Today, 16, 294-295, 1995).
[0004]
[Problems to be solved by the invention]
The inventors of the present invention synthesized various PDMP intermediates and analogs and confirmed their effects on glycolipid biosynthesis. Compared to PDMP analogs that have been confirmed to be active in the past, the present inventors have found that the amino acids of the present invention The present inventors have found that an alcohol derivative exhibits excellent activity and have reached the present invention. The present invention relates to a pharmaceutical containing an amino alcohol derivative or a pharmaceutically acceptable salt thereof, particularly in a pathological state in which cells grow abnormally in an undifferentiated state, a specific glycolipid molecule is contained in normal cells. Although it is abnormally expressed compared to, a drug that treats a disease based on abnormal growth of cells by causing the cells to leave the abnormal growth state by suppressing the expression of such glycolipids, and to differentiate into a normal state, Furthermore, it aims at providing a cancer therapeutic agent.
[0005]
[Means for Solving the Problems]
That is, the gist of the present invention is the general formula (I)
    [Chemical formula 3]
  R²-CH₂-C * H (NHP¹) -C * H (OR^Three-R¹・ ・ ・ ・ ・ ・ ・ (I)
(In the formula, * represents an asymmetric carbon and R¹IsPhenylRepresents the group R²IsPyrrolidino, piperidino or piperazino group optionally substituted by hydroxyl, methyl or hydroxymethyl group, or morpholinoRepresents the group R^ThreeIs a hydrogen fieldChildP¹Is4 to 16 carbon atomsAn alkyl group,6-18 carbon atomsAn aralkyloxycarbonyl group,4 to 16 carbon atomsAn alkoxycarbonyl group,Or 4-16 carbon atomsAlkenyloxycarbonylGroupTo express. )
Or a pharmaceutically acceptable salt thereof.
[0006]
Another gist of the present invention is the general formula (I).
      [Chemical formula 4]
    R²-CH₂-C * H (NHP¹) -C * H (OR^Three-R¹ ・ ・ ・ ・ ・ ・ (I)
(In the formula, * represents an asymmetric carbon and R¹IsPhenylRepresents the group R²IsPyrrolidino, piperidino or piperazino group optionally substituted by hydroxyl, methyl or hydroxymethyl group, or morpholinoRepresents the group R^ThreeIs a hydrogen fieldChildP¹Is4 to 16 carbon atomsAn alkyl group,6-18 carbon atomsAn aralkyloxycarbonyl group,4 to 16 carbon atomsAn alkoxycarbonyl group,Or 4-16 carbon atomsAlkenyloxycarbonylGroupTo express. It exists in the therapeutic agent for the abnormal proliferative disease containing the amino alcohol derivative shown by this.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in detail.
The present invention relates to a pharmaceutical comprising as an active ingredient an aminoalcohol derivative that is a glycosphingoglycolipid biosynthesis regulator, or a pharmaceutically acceptable salt thereof, as well as an aminoalcohol derivative exhibiting glycolipid biosynthesis inhibitory activity or a pharmaceutical product thereof The present invention relates to a therapeutic agent for abnormally proliferative diseases, particularly cancer, comprising a salt acceptable as an active ingredient. Here, “glucosphingoglycolipid” means glucosylceramide and glycosphingolipid biosynthesized using it as a starting material.
In the present invention, “lower” means having 1 to 6 carbon atoms.
[0008]
The amino alcohol derivative used as an active ingredient of the medicament and the therapeutic agent for abnormal proliferative disease of the present invention is a compound represented by the above general formula (I). In general formula (I), R¹Represents an aryl group, preferably a phenyl group which may be substituted with 1 to 3 identical or different substituents selected from lower alkyl, lower alkoxy, hydroxyl, hydroxy lower alkyl and nitro, more preferably Is a phenyl group. R²Is a cyclic amino group, preferably a pyrrolidino, piperidino or piperazino group which may have a hydroxyl, lower alkyl or hydroxy lower alkyl group, or a morpholino group, such as pyrrolidino, hydroxypyrrolidino, hydroxymethylpyrrolidino , Piperidino, hydroxypiperidino, piperazino, N-methylpiperazino or morpholino groups, more preferably cyclic amino groups such as pyrrolidino, hydroxypyrrolidino, piperidino, hydroxypiperidino or morpholino groups, most preferred Is a pyrrolidino group.
[0009]
R^ThreeIs a hydrogen or acyl group, preferably a lower acyl group which may have a substituent such as hydrogen, acetyl group, succinyl group, N, N-dimethylaminoacetyl group or methoxyacetyl group, more preferably hydrogen. is there. P¹Represents an alkyl group, an aralkyloxycarbonyl group, an alkoxycarbonyl group, an alkenyloxycarbonyl group, a hydroxyalkylcarbonyl group or an alkylcarbonyl group having 4 to 8 carbon atoms, preferably an alkyl group having 4 to 16 carbon atoms, or 6 to 6 carbon atoms. An aralkyloxycarbonyl group having 18 to 16 carbon atoms, an alkoxycarbonyl group having 4 to 16 carbon atoms, an alkenyloxycarbonyl group having 4 to 16 carbon atoms, a hydroxyalkylcarbonyl group having 4 to 16 carbon atoms, or an alkylcarbonyl group having 4 to 8 carbon atoms. More preferably a benzyloxycarbonyl group, a hydroxyalkylcarbonyl group having 6 to 8 carbon atoms, or an alkylcarbonyl group having 6 to 8 carbon atoms.
[0010]
The pharmaceutical and therapeutic agent for abnormal proliferative disease containing the amino alcohol derivative represented by the general formula (I) of the present invention as an active ingredient are represented by the formula R¹Is a phenyl group optionally substituted with 1 to 3 identical or different substituents selected from lower alkyl, lower alkoxy, hydroxyl, hydroxy lower alkyl and nitro, and R²Is a pyrrolidino, hydroxypyrrolidino, piperidino, hydroxypiperidino or morpholino group and R^ThreeAn amino alcohol derivative in which is hydrogen or a lower acyl group, or a pharmaceutically acceptable salt thereof.
Furthermore, the therapeutic agent for abnormal proliferative diseases is represented by the general formula (I):¹Is a phenyl group and R²Is a pyrrolidino group and R^ThreeIs hydrogen and P¹Is a benzyloxycarbonyl group, a hydroxyalkylcarbonyl group having 6 to 8 carbon atoms, or an alkylcarbonyl group having 6 to 8 carbon atoms, particularly an aminoalcohol derivative having a configuration of (1R, 2R) or a pharmaceutically acceptable derivative thereof Salt as an active ingredient.
[0011]
Among the aminoalcohol derivative compounds of the present invention, a compound whose steric configuration is (1R, 2R) in the general formula (I) has high glycolipid biosynthesis inhibitory activity, and further R in the general formula (I).¹Is a phenyl group and R²Is a pyrrolidino group and R^ThreeIs hydrogen and P¹Is a benzyloxycarbonyl group, and the configuration thereof is (1R, 2R) has a significantly higher activity than PDMP and its analogs, which currently have glycolipid biosynthesis inhibitory activity. Low toxicity was confirmed.
[0012]
The amino alcohol derivative of the present invention can be synthesized by the method shown in the following synthesis routes 1 to 3.
[Synthesis route 1]
Treatment of optically active N-protected-2-aminoalkanediol represented by compound a with methanesulfonyl chloride, for example, in the presence of pyridine in a solvent such as pyridine or in an anhydrous solvent such as dichloromethane, from ice cooling to room temperature. Then, the leaving group (Y) is introduced only into the primary hydroxyl group of the diol (for example, methanesulfonylation (mesylation)) to synthesize the compound b. In the formula, Y represents a leaving group, and specifically represents methanesulfonyl (mesyl), trihalogenomethanesulfonyl such as trifluoromethanesulfonyl, p-toluenesulfonyl, benzenesulfonyl, p-bromobenzenesulfonyl group and the like. After isolation of this product (compound b), in some cases without isolation, the starting material is used as it is in an organic solvent such as ethyl alcohol or N, N-dimethylformamide.²Treatment with an amine represented by H can lead to a compound represented by the general formula (I) -1. This amine is R²H, where R²Represents a cyclic amino group. Further, if necessary, the hydroxyl group of the obtained compound of formula (I) -1 is acylated (esterified) by a method known per se to give a compound of general formula (I) -2 (provided that R^ThreeCan lead to a compound represented by (acyl group).
The esterification reaction is usually performed at about 0 to 50 ° C., preferably at room temperature (5-35 ° C. (JIS K0050)) for several hours to several days, preferably 11 hours to 18 hours. If so, it can be appropriately set by a preliminary experiment.
[0013]
      [Synthesis Route 2] P in general formula (I)¹Is an alkoxycarbonyl group, an alkenyloxycarbonyl group, a hydroxyalkylcarbonyl group, or an alkylcarbonyl group (in the following description, P¹R^FourCO can be synthesized according to the synthesis route 2 using the compound represented by the general formula (I) -1 as a starting material. That is, the amino protecting group (P¹), For example, an aralkyloxycarbonyl group such as a benzyloxycarbonyl group is removed by a conventional deprotection method such as catalytic reduction, acid treatment, base treatment, etc., leading to compound c.
  The resulting amino group is then represented by the formulaR ^Four Amino acid represented by the general formula (I) -3 is obtained by acylation (amidation) with a carboxylic acid represented by COOH or a reactive derivative thereof such as an acid halide or acid anhydride of the carboxylic acid. Can lead to alcohol derivatives. In this formula, R^FourCO- represents an alkoxycarbonyl group having 4 to 16 carbon atoms, an alkenyloxycarbonyl group having 4 to 16 carbon atoms, a hydroxyalkylcarbonyl group having 4 to 16 carbon atoms, or an alkylcarbonyl group having 4 to 8 carbon atoms. Specifically, the acyl group (R^FourIn the case where the carbon atom of CO-) is an alkylcarbonyl group having 6 carbon atoms, hexanoyl chloride or hexanoic anhydride is used as the acylating agent.
[0014]
In addition, compound c is converted to carboxylic acid (R^FourCOOH) and condensing agents commonly used in amide bond reactions can also be reacted to the desired amino alcohol derivatives. Specifically, fatty acids such as hexanoic acid and 2-hydroxyoctanoic acid or hydroxy-substituted fatty acids are used as the carboxylic acid, and known ones such as dicyclohexylcarbodiimide (DCC) and water-soluble carbodiimide are used as the condensing agent. Specific examples of the water-soluble carbodiimide include 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (EDC).
Further, if necessary, the hydroxyl group of the compound of the formula (I) -3 is acylated by a method known per se in the same manner as in the synthesis route 1, and the compound of the general formula (I) -4^ThreeCan lead to a compound represented by (acyl group).
[0015]
[Synthesis route 3]
In general formula (I), P¹A compound of the general formula (I) -5 in which is an alkyl group (in the following description, P¹R^FiveCH₂(Usually having 4 to 16 carbon atoms) can be synthesized by the synthesis route 3. That is, a compound represented by compound d (wherein R^FiveCO represents an alkylcarbonyl group having 4 to 16 carbon atoms) and is treated with a reducing agent such as lithium aluminum hydride to lead to the target amino alcohol derivative represented by the general formula (I) -5. it can.
Further, if necessary, the hydroxyl group at the 3-position of the compound of the formula (I) -5 is acylated (esterified) by a method known per se in the same manner as in the synthesis route 1 to give a compound of the general formula (I) -6 (where R^ThreeCan lead to a compound represented by (acyl group).
[0016]
(Isolation, purification)
Isolation / purification of the compound of the present invention obtained by each of the above synthesis routes is known after the reaction, solvent extraction with ethyl acetate, chloroform, etc., various chromatography (adsorption chromatography, ion exchange chromatography, etc.), crystallization itself. The purification means can be combined appropriately.
[0017]
Synthesis route 1
[Chemical formula 5]

[0018]
Synthesis route 2
[Chemical 6]

[0019]
Synthesis route 3
[Chemical 7]

[0020]
As a method for synthesizing N-protected-2-aminoalkanediols which are starting materials of the above synthesis route 1, a method of reducing N-protected-α-aminoketones (J. Org. Chem., 54, 1866 (1989)). )), N- (diphenylmethylene) amino acid ester treated with diisobutylaluminum hydride followed by Grignard reagent (J. Org. Chem., 57, 5469 (1992)), N-protected amino aldehydes and N- Protection-Method of reacting an acid chloride of an amino acid with an organometallic reagent (J. Am. Chem. Soc., 95, 4098 (1973)), Asymmetric aldol reaction of 2-oxazolidinone and an aldehyde (J. Am. Chem. Soc., 108, 6757 (1986)) (Evans method), chiral imidazolidinone and asymmetric aldol reaction of oxazolidinone and aldehyde (Helv. Chem. Acta, 70, 237 (1987)) and the like are known.
On the other hand, as a method for synthesizing the N-protected-α-amino ketones, a method in which a carboxyl group of an amino acid is converted into an acid chloride using N-protected-α-amino acid as a starting material, and then reacted with benzene and Friedel-Crafts reaction ( J. Am. Chem. Soc., 103, 6157 (1981)), a method in which the carboxyl group of the amino acid is treated with an alkyl lithium reagent to form a lithium salt, and then reacted with a Grignard reagent (J. Am. Org. Chem., 54, 1866 (1989)).
[0021]
Pharmaceutically acceptable salts of the compound represented by formula (I) include inorganic acid salts such as hydrochloric acid, phosphoric acid, sulfuric acid and nitric acid, formic acid, acetic acid, citric acid, lactic acid, malic acid, oxalic acid and maleic acid. Examples thereof include salts of organic acids such as acid, fumaric acid, succinic acid, trifluoroacetic acid, methanesulfonic acid (mesyl acid), and P-toluenesulfonic acid. Such a salt can be produced by a method known per se. For example, the compound represented by the general formula (I) (free form) is dissolved in an appropriate solvent such as alcohol, and the above-mentioned acid is usually equimolar. May be added and reacted, and the solvent may be distilled off if desired.
The solubility of the compound of the present invention in water or physiological saline is improved when the compound of the present invention is in a salt form such as hydrochloride, citrate, lactate, succinate, etc. than in the free form.
[0022]
[Action]
The amino alcohol derivative of the present invention represented by the general formula (I) or a pharmaceutically acceptable salt thereof (hereinafter sometimes referred to as “the compound of the present invention”) has an action of controlling the biosynthesis of glycolipid. , It has utility as a medicine based on this action.
The compound represented by the general formula (I) has asymmetric carbons in at least two positions in the molecule, and each isomer by its configuration (L-threo, L-erythro, D-threo, D-erythro) Each of these biosynthesis control actions is different. In particular, among the compounds of the present invention, a compound having an inhibitory action on glycolipid biosynthesis has an action of inducing differentiation of cells that proliferate abnormally in an undifferentiated state or an action of normalizing abnormally proliferating cells, and is abnormally proliferative. It is useful as a therapeutic agent for diseases, particularly as a therapeutic agent for cancer. The compound of the present invention preferable for such use has a steric configuration of D-threo or L-threo, and D-threo is particularly preferable.
In addition, the configuration of the compound of the general formula (I), (1S, 2S), (1R, 2S), (1S, 2R) or (1R, 2R) are respectively an L-threo isomer, an L-erythro isomer, D -Corresponds to erythro or D-threo form.
[0023]
Since the compound of the present invention has an action of inducing differentiation of undifferentiated and abnormally proliferating cells, the therapeutic agent of the present invention exhibits abnormal proliferative diseases in mammals including humans, that is, abnormal proliferation in the state of undifferentiated cells. It is used as a pharmaceutical for the treatment of various diseases based on presenting, that is, for the purpose of treatment, alleviation (amelioration of symptoms), maintenance (prevention of deterioration) or prevention of such diseases. Such diseases include benign tumors (such as uterine fibroids); solid cancers (such as canine cancer, colon cancer, lung cancer, stomach cancer, pancreatic cancer, squamous cell carcinoma such as pancreatic cancer, liver cancer, or adenocarcinoma, or brain tumor), glioma, Malignant tumor or cancer (carcinoma, sarcoma) such as leukemia and malignant lymphoma; nephritis (glomerulonephritis, etc.), human autoimmune lymphoproliferative syndrome, lymphoproliferative disorder, angioimmunoblastic lymphadenopathy, immunity Blastic lymphadenopathy, systemic lupus erythematosus, inflammatory bowel disease (Crohn's disease, ulcerative colitis, etc.), progressive systemic sclerosis, polymyositis (dermatomyositis), Sjogren's syndrome, myelodysplastic syndrome, strong Although abnormal proliferative diseases such as dermatosis, rheumatism, psoriasis, and wound hyperplasia (such as granulation) are exemplified, the therapeutic agent of the present invention is preferably effective for the treatment of malignant tumors, particularly solid cancers.
[0024]
[Formulation]
The compound of the present invention can be made into a preparation administered orally or parenterally together with a carrier, an excipient, and other additives.
Examples of oral preparations include solid preparations such as powders, granules, capsules and tablets; liquid preparations such as syrups, elixirs and emulsions. The powder can be obtained by mixing with excipients such as lactose, starch, crystalline cellulose, calcium lactate, calcium hydrogen phosphate, magnesium aluminate metasilicate, and silicic anhydride. In addition to the above-mentioned excipients, granules may be prepared in a wet or dry manner by further adding, for example, a binder such as sucrose, hydroxypropylcellulose, polyvinylpyrrolidone, or a disintegrant such as carboxymethylcellulose or carboxymethylcellulose calcium. Can be obtained by granulation. Tablets can be obtained by compressing the powder or granules as described above or by adding a lubricant such as magnesium stearate or talc. The tablet or granule is coated with an enteric base such as hydroxypropylmethylcellulose phthalate, methyl methacrylate copolymer, hydroxypropylmethylcellulose acetate, hydroxypropylmethylcellulose succinate, or ethylcellulose, carnauba wax, hydrogenated oil, white shellac, etc. These can be made into enteric or persistent formulations. Among capsules, a hard capsule can be obtained by filling the above powder or granule into a hard capsule, and a soft capsule dissolves the compound of the present invention in glycerin, polyethylene glycol, sesame oil, olive oil, etc. This can be obtained by coating with a gelatin film.
[0025]
A syrup can be obtained by dissolving a sweetener such as sucrose, sorbitol, glycerin and the compound of the present invention in water. In addition to sweeteners and water, essential oils, ethanol, etc. are added to make elixirs, or Arabia gum, tragacanth, polysorbates (polysorbate 20, polysorbate 60, polysorbate 80 (Tween 80), etc.), sodium carboxymethylcellulose, etc. Can be added to form an emulsion or suspension. Moreover, a corrigent, a coloring agent, a preservative, etc. can be added to these liquid preparations as needed.
[0026]
Examples of parenteral preparations include injections, rectal administration agents, pessaries, external preparations for skin, inhalants, aerosols, eye drops and the like. Injections include compounds of the present invention and nonionic surfactants such as polysorbates as necessary; pH adjusters such as hydrochloric acid, sodium hydroxide, lactic acid, sodium lactate, sodium monohydrogen phosphate, sodium dihydrogen phosphate An isotonic agent such as sodium chloride and glucose; a stabilizer such as amino acids; and distilled water for injection or physiological saline; Further, mannitol, dextran, gelatin and the like can be added and lyophilized in vacuum to obtain a dissolution type injection for use. In addition, it can be a powder-filled injection. Moreover, after adding an emulsifier, such as a lecithin, polysorbates, polyoxyethylene hydrogenated castor oil, macrogol, to this invention compound, it can also be set as the emulsion for injection emulsified in water.
[0027]
Examples of injections include liposome preparations and lipid microspheres that can improve solubility and the rate of transfer to target organs. Liposome preparation is a known method for preparing liposomes (CG Knight, Liposomes: From Physical Structure to Therapeutic Applications, pp. 51-82, Elsevier, Amsterdam (1981); Proc. Natl. Acad. Sci., USA, Vol. 75, 4194. (1978)).
[0028]
That is, examples of amphipathic substances that form liposome membranes include natural phospholipids (egg yolk lecithin, soybean lecithin, sphingomyelin, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, diphosphatidylglycerol, phosphatidylethanolamine, cardiolipin). Phospholipids such as lipids (distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, etc.) are used. It is also known to impart negative charges to cholesterols (cholesterol, ergosterol, phytosterol, sitosterol, stigmasterol, etc.) and liposomes in order to improve membrane stability, fluidity, and drug membrane permeability. Substances (phosphatidic acid, dicetyl phosphate, etc.), substances known to impart a positive charge (stearylamine, stearylamine acetate, etc.), antioxidants (tocopherol, etc.), oily substances (soybean oil, cottonseed oil) , Sesame oil, liver oil, etc.) may be used.
[0029]
The liposome can be produced, for example, by the following method. The above amphiphile and additive and the compound of the present invention are each dissolved in an organic solvent (single or mixed solvent such as chloroform, dichloromethane, ethanol, methanol, hexane, etc.), and both solutions are mixed together in a container such as a flask. The organic solvent is removed in the presence of an inert gas (nitrogen gas, argon gas, etc.) and a thin film is deposited on the vessel wall. Next, this thin film is added to an appropriate aqueous medium (physiological saline, buffer solution, phosphate buffered physiological saline, etc.) and stirred with a stirrer. In order to obtain a liposome having a small particle size, it is further dispersed using an ultrasonic emulsifier, a pressure emulsifier, a French press cell crusher, or the like. In this way, a solution in which an amphiphile necessary for liposome formation and the compound of the present invention are dispersed in an aqueous medium is subjected to membrane filter treatment, whereby liposome formation proceeds, and the nanosphere-liposome with controlled particle size distribution (ultralipid Fine particles; particle size of about 25 to 50 nm) can be obtained. Further, the liposome may be subjected to fractionation treatment such as ultrafiltration, centrifugation, and gel filtration to remove the unsupported drug.
[0030]
Moreover, it is obtained by adding β-octylglucoside, L-tyrosine-7-amido-4-methylcoumarin, phenylamino mannoside or sulfatide as a film-forming substance in addition to the above-mentioned amphipathic substances and additives. The amino alcohol derivative of the general formula (I) of the present invention can be supported on a liposome having a glucose residue, a tyrosine residue, a mannose residue or a sulfatide on the membrane (the method itself is disclosed in JP-A-4-69332). Issue).
[0031]
Lipid microspheres are prepared by dissolving the compound of the present invention in soybean oil, sesame oil, etc., adding natural phospholipids, glycerin, water, etc., and stirring with a stirrer. Further, ultrasonic emulsifier, pressure emulsifier, French press cell crusher It can be obtained by dispersing using, for example.
[0032]
For rectal administration, a suppository base such as mono-, di- or triglyceride of cacao fatty acid, polyethylene glycol, etc. is added to the compound of the present invention, and then heated and melted, poured into a mold and cooled, Alternatively, the compound of the present invention can be obtained by dissolving in polyethylene glycol, soybean oil or the like and then coating with a gelatin film.
The external preparation for skin can be obtained by adding white petrolatum, beeswax, liquid paraffin, polyethylene glycol or the like to the compound of the present invention, heating as necessary, and kneading.
The tape agent can be obtained by kneading a pressure-sensitive adhesive such as rosin or alkyl acrylate polymer in the compound of the present invention and spreading it on a nonwoven fabric or the like.
The inhalant can be obtained by, for example, dissolving or dispersing the compound of the present invention in a propellant such as a pharmaceutically acceptable inert gas and filling it in a pressure resistant container.
[0033]
[Method of administration]
The administration method of the pharmaceutical comprising the compound of the present invention as an active ingredient is not particularly limited, but particularly when used for the treatment of abnormal proliferative diseases, intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, etc. Rectal administration, transpulmonary administration and the like are preferred. As a method for treating an abnormal proliferative disease, a method of direct administration to a disease site (local injection, etc.) can be mentioned.
The dose is appropriately determined according to the patient's age, health condition, body weight, etc., but is generally 0.25 to 200 mg / kg, preferably 0.5 to 100 mg / kg once or more a day. Divide and administer.
〔toxicity〕
The aminoalcohol derivative represented by the general formula (I), which is an active ingredient of the medicament of the present invention, exhibits little or no cytotoxicity at doses exhibiting pharmacological activity.
[0034]
【Example】
EXAMPLES Next, although an Example demonstrates this invention still in detail, this invention is not limited to a following example, unless the summary is exceeded.
Production methods of the compound of the present invention and synthetic intermediates thereof are shown as preparation examples.
In the following examples, MeOH, AcOEt, AcOH, and DMF represent methanol, ethyl acetate, acetic acid, and N, N-dimethylformamide, respectively.
[0035]
Preparation Example 1 Synthesis of (1S, 2S) -2-benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester
(1S, 2S) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol (21.2 g, 70.3 mmol) was dissolved in pyridine (350 ml), and methanesulfonyl chloride (5. 6 ml, 72.3 mmol) was added dropwise over 5 minutes. After stirring for 30 minutes on an ice bath, the mixture was stirred overnight at room temperature. After confirming the completion of the reaction by TLC (chloroform: methanol = 20: 1), the solvent was distilled off, ethyl acetate (500 ml) was added, 1N-HCl (250 ml × 3), saturated brine ( 250 ml) and dried over sodium sulfate, and the solvent was distilled off. The precipitated crystals were washed with ethyl acetate: hexane = 1: 1 to obtain the title material (25.3 g, yield 95.0%) as white crystals.
TLC Rf 0.55 (CHCl_Three: MeOH = 20: 1), 0.83 (AcOEt), 0.62 (Hexane: AcOEt = 1: 2)
¹H-NMR (CDCl_Three) δ: 7.35-7.26 (10H, m, aromatic), 5.30 (1H, d, J = 7.81Hz, NH), 5.02 (2H, s, CH ₂-O-CO), 4.99 (1H, d, J = 3.91Hz, CH-OH), 4.43-4.39,4.22-4.12 (3H, m, N-CH-CH ₂), 2.98 (3H, s, SO_ThreeCH_Three)
In addition, instead of (1S, 2S) -2-benzyloxycarbonylamino-1-phenyl-1,3-propanediol, (1R, 2R) -2-benzyloxycarbonylamino-1-phenyl-1,3- By using propanediol, (1R, 2R) -2-benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester could be synthesized.
[0036]
Preparation Example 2 Synthesis of (1R, 2R) -2-benzyloxycarbonylamino-3-morpholino-1-phenyl-1-propanol
(1R, 2R) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (1.21 g, 3.19 mmol) was dissolved in N, N-dimethylformamide (6 ml). At room temperature, morpholine (1.11 g, 12.8 mmol) was added, and the mixture was stirred at 40 ° C. for 24 hours. After confirming that the reaction was almost complete by TLC (chloroform: methanol = 20: 1, hexane: ethyl acetate = 1: 2, ethyl acetate), saturated sodium hydrogen carbonate solution (70 ml), ethyl acetate (100 ml) The organic layer was washed successively with water and saturated brine, dried over sodium sulfate and filtered. The solvent was distilled off, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 2) to obtain the title material (507.5 mg, yield 43.0%) as a colorless oil.
TLC Rf 0.32 (CHCl_Three: MeOH = 20: 1), 0.12 (Hexane: AcOEt = 1: 2)
¹H-NMR (CDCl_Three) δ: 7.38-7.26 (10H, m, aromatic), 5.04 (2H, s, CH₂O-CO), 5.00 (1H, d, J = 3.41Hz, H-1), 4.11 (1H, m, H-2), 3.72 (4H, m, (CH₂)₂O), 2.68-2.47 (6H, m, (CH₂)_ThreeN)
[0037]
Preparation Example 3 Synthesis of (1R, 2R) -2-benzyloxycarbonylamino-3-pyrrolidino-1-phenyl-1-propanol
(1R, 2R) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (1.52 g, 4.01 mmol) was dissolved in DMF (8 ml) and pyrrolidine (1. 14 g, 16.03 mmol) was added, and the mixture was stirred at 40-50 ° C. for 18 hours. Ethyl acetate (100 ml) was added, and the organic layer was saturated sodium hydrogen carbonate solution (70 ml), water (70 ml), saturated brine (70 ml). ) And then dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting crude product was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain the title material (1.21 g, yield 85.5%) as a colorless oil.
TLC Rf 0.20 (CHCl_Three: MeOH = 20: 1), 0.20 (AcOEt)
¹H-NMR (CDCl_Three) δ: 7.39-7.24 (10H, m, aromatic), 5.06-5.02 (2H, m, CH₂-O-CO), 4.99 (1H, d, J = 3.91Hz, H-1), 4.07 (1H, m, H-2), 2.9-2.6 (6H, m, (CH₂)_ThreeN), 1.83-1.74 (4H, m, H-3 ', H-4')
¹³C-NMR (CDCl_Three) δ: 156.0, 140.8, 136.5, 128.4, 128.2, 128.0, 127.8, 127.4, 126.1, 75.7, 66.6, 58.1, 55.2, 53.4, 23.6
[0038]
Moreover, the hydrochloride of this compound was prepared by the following method.
(1R, 2R) -2-Benzyloxycarbonylamino-3-pyrrolidino-1-phenyl-1-propanol (95.1 mg, 0.269 mmol) was dissolved in ethanol (3 ml), and 2N hydrochloric acid (161. 2 μl, 0.322 mmol) was added, and after stirring for 10 minutes, the solvent was distilled off under reduced pressure. Thereafter, the operation of adding ethanol (3 ml) and evaporating under reduced pressure was repeated three times, followed by drying under reduced pressure at room temperature for 16 hours to give (1R, 2R) -2-benzyloxycarbonylamino-3-pyrrolidino-1-like pale yellow oil. 105.5 mg (100% yield) of phenyl-1-propanol hydrochloride was obtained.
[0039]
Preparation Example 4 Synthesis of (1R, 2R) -2-benzyloxycarbonylamino-3-piperidino-1-phenyl-1-propanol
(1R, 2R) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (1.21 g, 3.19 mmol) was dissolved in DMF (6 ml) and piperidine (1. (09 g, 12.8 mmol) was added, and the mixture was stirred at 40 to 50 ° C. for 24 hours. After confirming that the reaction was almost complete by TLC (ethyl acetate, chloroform: methanol = 20: 1), ethyl acetate (100 ml) was added, and the organic layer was saturated sodium bicarbonate solution (70 ml), water (70 ml). ), Saturated brine (70 ml), and then dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting crude product was purified by silica gel column chromatography (ethyl acetate) to obtain the title material (795.4 mg, yield 68.0%) as a colorless oil.
TLC Rf 0.20 (CHCl_Three: MeOH = 20: 1), 0.17 (AcOEt)
¹H-NMR (CDCl_Three) δ: 7.36-7.25 (10H, m, aromatic), 5.04 (2H, s, CH₂-O-CO), 5.01 (1H, d, J = 3.42Hz, H-1), 4.94 (1H, d, J = 7.33Hz, NH), 4.15 (1H, m, H-2), 2.64-2.45 (6H, m, (CH₂)_ThreeN), 1.68-1.54 (4H, m, H-3 ', H-5'), 1.5-1.4 (2H, m, H-4 ')
¹³C-NMR (CDCl_Three) δ: 155.9, 140.8, 136.4, 128.5, 128.3, 128.1, 127.9, 127.4, 126.3, 75.7, 66.7, 60.5, 55.8, 51.7, 26.1, 23.9
[0040]
Preparation Example 5 Synthesis of (1S, 2S) -2-t-butoxycarbonylamino-3-morpholino-1-phenyl-1-propanol
(1S, 2S) -2-t-Butoxycarbonylamino-1-phenyl-1,3-propanediol was mesylated according to the method of Preparation Example 1 and then subjected to a morpholine substitution reaction according to the method of Preparation Example 2. The title material was obtained as a colorless oil in a yield of 63%.
TLC Rf 0.36 (CHCl_Three: MeOH = 20: 1)
¹H-NMR (CDCl_Three) δ: 7.38-7.26 (5H, m, aromatic), 4.98 (1H, d, J = 3.91Hz, H-1), 4.05 (1H, m, H-2), 3.74 (4H, m, (CH₂)₂O), 2.64-2.59 (5H, m, H-2 ', H-6', H-3A), 2.46 (1H, dd, J = 4.89,13.19Hz, H-3B), 1.38 (9H, s, (CH_Three)_Three)
[0041]
Preparation Example 6 Synthesis of (1S, 2S) -2-benzyloxycarbonylamino-3- (N-methylpiperazino) -1-phenyl-1-propanol
(1S, 2S) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (1.81 g, 4.78 mmol) was dissolved in ethanol (40 ml) and sodium iodide ( (712.8 mg, 4.75 mmol) and N-methylpiperazine (1.92 g, 19.2 mmol) were added, and the mixture was stirred at 50 ° C. for 5 days. After confirming the reaction status by TLC (chloroform: methanol = 9: 1), the solvent was distilled off, water (50 ml) and ethyl acetate (100 ml) were added, and the organic layer was washed successively with water and saturated brine, Dried over sodium sulfate and filtered. The solvent was distilled off, and the residue was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain the title material (242.2 mg, yield 13.2%) as a colorless oil.
TLC Rf 0.38 (CHCl_Three: MeOH = 9: 1)
¹H-NMR (CDCl_Three) δ: 7.36-7.26 (10H, m, aromatic), 5.04 (2H, s, CH₂-O-CO), 5.00 (1H, d, J = 3.41Hz, H-1), 4.97 (1H, d, NH), 4.12 (1H, m, H-2), 2.70-2.49 (10H, m, (CH₂)_ThreeN, (CH₂)₂N), 2.28 (3H, s, CH_Three-N)
¹³C-NMR (CDCl_Three) δ: 156.0, 140.7, 136.4, 128.5, 128.3, 128.1, 127.9, 127.5, 126.2, 75.3, 66.8, 59.6, 55.1, 54.1, 52.1, 45.9
[0042]
Preparation Example 7 Synthesis of (1S, 2S) -2-benzyloxycarbonylamino-3-((2S) -2-hydroxymethylpyrrolidino) -1-phenyl-1-propanol
(2S) -2-hydroxymethylpyrrolidine (323.3 mg, 3.20 mmol) was dissolved in ethanol (12 ml), and (1S, 2S) -2-benzyloxycarbonylamino-1-phenyl-1,3-propanediol- The solution was added dropwise to a methylene chloride solution (3 ml) of 3-p-bromobenzenesulfonyl ester (782 mg, 1.50 mmol). After stirring at 45 ° C. for 2 days, the reaction status was confirmed by TLC (chloroform: methanol = 9: 1, ethyl acetate: 2-propanol = 2: 1), the solvent was distilled off, and the residue was subjected to silica gel column chromatography ( Purification with ethyl acetate: 2-propanol = 7: 3) gave the title material (79.5 mg, yield 13.8%) as a colorless oil.
TLC Rf 0.25 (CHCl_Three: MeOH = 9: 1), 0.39 (AcOEt: (CH_Three)₂(CHOH = 2: 1)
¹H-NMR (CDCl_Three) δ: 7.51-7.23 (10H, m, aromatic), 5.32 (1H, br, NH), 4.99 (3H, m, H-1, CH₂-O-CO), 3.93 (1H, m, H-2), 3.67,3.66,3.64,3.63 (1H, dd, CH ₂(A) -OH), 3.51 (1H, dd, J = 4.40,11.23Hz, CH ₂(B) -OH), 3.28-3.23 (1H, m, H-5'A), 3.08 (1H, dd, J = 5.86,13.19Hz, H-3A), 2.81 (1H, dd, J = 2.93, 13.18Hz, H-3B), 2.71 (1H, m, H-2 '), 2.34-2.28 (1H, m, H-5'B), 1.90-1.59 (4H, m, H-3', H- Four')
¹³C-NMR (CDCl_Three) δ: 156.5, 141.0, 136.5, 128.4, 128.3, 128.0, 127.8, 127.5, 125.8, 75.4, 66.6, 66.4, 63.7, 58.0, 56.2, 55.4, 27.0, 23.8
[0043]
Preparation Example 8 Synthesis of (1S, 2S) -2-benzyloxycarbonylamino-3- (3-hydroxypyrrolidino) -1-phenyl-1-propanol
(1S, 2S) -2-benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (2.60 g, 6.86 mmol) was dissolved in ethanol (20 ml) to give 3-hydroxypyrrolidine. (1.19 g, 13.68 mmol) was added, and the mixture was stirred at 45 ° C. for 5 days. After confirming the reaction status by TLC (chloroform: methanol = 9: 1), the solvent was distilled off, and the residue was purified by silica gel column chromatography (chloroform: methanol = 9: 1, ethyl acetate: methanol = 9: 1). This gave the title material (527.1 mg, yield 20.8%) as a colorless oil.
TLC Rf 0.25 (CHCl_Three: MeOH = 9: 1), 0.35 (AcOEt: MeOH = 4: 1)
¹H-NMR (CDCl_Three) δ: 7.41-7.24 (10H, m, aromatic), 5.26 (0.7H, d, J = 7.82Hz, NH, derived from one diastereomer), 5.20 (0.3H, d, NH, other diastereomer) 5.00 (3H, s, H-1, CH₂-O-CO), 4.34 (0.7H, m, H-3 ', derived from one diastereomer), 4.28 (0.3H, m, H-3', derived from the other diastereomer), 4.02 (1H , m, H-2), 3.04-2.99, 2.89-2.42 (6H, m, (CH₂)_ThreeN), 2.20-2.07 (1H, m, H-4'A), 1.80-1.68 (1H, m, H-4'B)
¹³C-NMR (CDCl_Three) δ: 156.5, 141.2, 141.1, 136.7, 128.8, 128.6, 128.3, 128.2, 127.8, 126.4, 75.4, 75.2, 71.3, 67.0, 64.0, 58.0, 54.2, 54.1, 53.8, 34.9
[0044]
Preparation Example 9 Synthesis of (1S, 2S) -2-benzyloxycarbonylamino-3-pyrrolidino-1-phenyl-1-propanol
(1S, 2S) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (1.21 g, 3.19 mmol) was dissolved in DMF (6 ml) and pyrrolidine (0. 91 g, 12.8 mmol) was added, and the mixture was stirred at 40 ° C. for 24 hours. Ethyl acetate (100 ml) was added, and the organic layer was washed with saturated sodium bicarbonate solution (70 ml), water (70 ml), and saturated brine (70 ml). After sequential washing, the organic layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting crude product was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain the title material (983.1 mg, yield 87.0%) as a colorless oil.
TLC Rf 0.20 (CHCl_Three: MeOH = 20: 1), 0.20 (AcOEt)
¹H-NMR (CDCl_Three) δ: 7.39-7.24 (10H, m, aromatic), 5.06-5.02 (2H, m, CH₂-O-CO), 4.99 (1H, d, J = 3.91Hz, H-1), 4.07 (1H, m, H-2), 2.9-2.6 (6H, m, (CH₂)_ThreeN), 1.83-1.74 (4H, m, H-3 ', H-4')
¹³C-NMR (CDCl_Three) δ: 156.0, 140.8, 136.5, 128.4, 128.2, 128.0, 127.8, 127.4, 126.1, 75.7, 66.6, 58.1, 55.2, 53.4, 23.6
[0045]
Preparation Example 10 Synthesis of (1S, 2S) -2-benzyloxycarbonylamino-3- (3-hydroxymethylpiperidino) -1-phenyl-1-propanol
(1S, 2S) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (2.43 g, 6.41 mmol) was dissolved in ethanol (20 ml) to give 3-hydroxymethyl. Piperidine (1.47 g, 12.78 mmol) was added, and the mixture was stirred at 45 ° C. for 5 days. After confirming the reaction status by TLC (chloroform: methanol = 9: 1), the solvent was distilled off, and the residue was purified by silica gel column chromatography (chloroform: methanol = 20: 1, ethyl acetate: methanol = 20: 1). The title material (293.3 mg, 11.5%) was obtained as a colorless oil.
TLC Rf 0.42 (CHCl_Three: MeOH = 9: 1), 0.16 (AcOEt: MeOH = 20: 1)
¹H-NMR (CDCl_Three) δ: 7.35-7.26 (10H, m, aromatic), 5.03 (2H, s, CH₂-O-CO), 4.994 (0.5H, d, J = 7.81Hz, H-1, from one diastereomer), 4.986 (0.5H, d, J = 8.30Hz, H-1, the other diastereomer) (From stereomer), 4.15-4.09 (1H, m, H-2), 3.56-3.45 (2H, m, CH ₂-OH), 3.00-2.91, 2.75, 2.25-2.00 (4H, m, H-2 ', H-6'), 2.65-2.59 (1H, m, H-3A), 2.49-2.45 (1H, m, H-3B), 1.82 (1H, m, H-3 '), 1.75-1.65, 1.63-1.53, 1.09-1.04 (4H, m, H-4', H-5 ')
¹³C-NMR (CDCl_Three) δ: 156.2, 156.1, 140.7, 136.4, 128.5, 128.3, 128.1, 127.9, 127.5, 126.2, 75.4, 75.3, 66.7, 65.7, 65.6, 60.4, 60.3, 60.2, 58.2, 57.5, 55.7, 55.1, 52.0, 38.8 , 38.7, 26.6, 24.7, 14.2
[0046]
Preparation Example 11 Synthesis of (1S, 2S) -2-benzyloxycarbonylamino-3- (4-hydroxypiperidino) -1-phenyl-1-propanol
(1S, 2S) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (273.6 mg, 0.722 mmol) was dissolved in ethanol (3 ml) and sodium iodide ( 119.2 mg, 0.795 mmol) and 4-hydroxypiperidine (171.5 mg, 1.70 mmol) were added, and the mixture was stirred at room temperature for 4 days. After confirming the reaction status by TLC (chloroform: methanol = 9: 1), 4-hydroxypiperidine (157.0 mg, 1.55 mmol) was added, and the mixture was further stirred at 45 ° C. for 2 days. The solvent was distilled off, and the residue was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain the title material (112.6 mg, yield 40.6%) as a colorless oil.
TLC Rf 0.24 (CHCl_Three: MeOH = 9: 1)
¹H-NMR (CDCl_Three) δ: 7.36-7.25 (10H, m, aromatic), 5.03 (3H, m, CH₂-O-CO, NH), 5.00 (1H, d, J = 2.93Hz, H-1), 4.11 (1H, m, H-2), 3.71 (1H, m, H-4 '), 2.91, 2.82 , 2.64, 2.48, 2.32 (6H, m, (CH₂)_ThreeN), 1.89 (2H, m, H-3'A, H-5'A), 1.64-1.56 (2H, m, H-3'B, H-5'B)
¹³C-NMR (CDCl_Three) δ: 156.1, 140.7, 136.4, 128.5, 128.3, 128.1, 127.9, 127.5, 126.2, 75.4, 66.9, 66.8, 66.7, 59.5, 52.2, 51.9, 34.4
[0047]
Preparation Example 12 Synthesis of (1R, 2R) -2-benzyloxycarbonylamino-3- (4-hydroxypiperidino) -1-phenyl-1-propanol
(1R, 2R) -2-Benzyloxycarbonylamino-1-phenyl-1,3-propanediol-3-methanesulfonyl ester (295.6 mg, 0.780 mmol) was dissolved in ethanol (3 ml) to give 4-hydroxypiperidine. (315.1 mg, 3.12 mmol) was added, and the mixture was stirred at 45 ° C. for 2 days. After confirming the reaction status by TLC (chloroform: methanol = 9: 1), the solvent was distilled off, and the residue was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to give the title material (179, colorless oil). 0.7 mg, yield 60.0%).
TLC Rf 0.24 (CHCl_Three: MeOH = 9: 1)
¹H-NMR (CDCl_Three) δ: 7.36-7.25 (10H, m, aromatic), 5.03 (3H, m, CH₂-O-CO, NH), 5.00 (1H, d, J = 2.93Hz, H-1), 4.11 (1H, m, H-2), 3.71 (1H, m, H-4 '), 2.91, 2.82 , 2.64, 2.48, 2.32 (6H, m, (CH₂)_ThreeN), 1.89 (2H, m, H-3'A, H-5'A), 1.64-1.56 (2H, m, H-3'B, H-5'B)
¹³C-NMR (CDCl_Three) δ: 156.1, 140.7, 136.4, 128.5, 128.3, 128.1, 127.9, 127.5, 126.2, 75.4, 66.9, 66.8, 66.7, 59.5, 52.2, 51.9, 34.4
[0048]
Preparation Example 13 Synthesis of (1R, 2R) -2-amino-3-pyrrolidino-1-phenyl-1-propanol
(1R, 2R) -2-Benzyloxycarbonylamino-3-pyrrolidino-1-phenyl-1-propanol (394.2 mg, 1.11 mmol) was dissolved in methanol (10 ml) and 10% palladium on carbon (124.8 mg, 10.6 mol%) was added, and the mixture was stirred at room temperature for 30 minutes in a hydrogen atmosphere. After confirming the completion of the reaction by TLC (chloroform: methanol = 7: 3 and ethyl acetate: methanol = 2: 1), palladium carbon was removed by filtration. The filtrate was concentrated to give the title material (250.2 mg) as a colorless oil.
[0049]
Preparation Example 14 Synthesis of (1R, 2R) -2-octyloxycarbonylamino-3-pyrrolidino-1-phenyl-1-propanol
(1R, 2R) -2-amino-3-pyrrolidino-1-phenyl-1-propanol (250.2 mg, 1.11 mmol) was dissolved in methanol (5 ml), and triethylamine (0.186 ml, 1.337 mmol) was dissolved at room temperature. ) Was added, followed by addition of chloroformic acid n-octyl ester (0.240 ml, 1.226 mmol) on an ice bath and stirring at room temperature. 90 minutes later, triethylamine (0.186 ml, 1.337 mmol) and chloroformic acid n-octyl ester (0.240 ml, 1.226 mmol) were added, and the mixture was stirred at room temperature. After 20 hours, the solvent was distilled off under reduced pressure, ethyl acetate (100 ml) was added, and the organic layer was washed successively with saturated sodium hydrogen carbonate solution, water and saturated saline 70 ml each, dried over sodium sulfate, filtered, The solvent was removed under reduced pressure. The obtained crude product was purified by silica gel column chromatography (elution solvent; chloroform: methanol = 20: 1) to obtain the title material (99.5 mg, yield 23.8%) as a colorless oil.
TLC Rf 0.28 (AcOEt: MeOH = 4: 1), 0.30 (CHCl_Three: MeOH = 9: 1)
¹H-NMR (CDCl_Three) δ: 7.36-7.24 (5H, m, aromatic), 5.05 (1H, d, J = 2.93Hz, H-1), 4.9 (1H, d, NH), 4.04 (1H, m, H-2), 3.96 (2H, m, COOCH₂), 2.91-2.68 (6H, m, CH₂N (CH₂)₂),
1.80 (4H, m, H-3 ', H-4'), 1.52 (2H, m, COOCH₂CH ₂), 1.26 (10H, m, (CH ₂)_FiveCH_Three), 0.88 (3H, t, CH₂CH _Three)
¹³C-NMR (CDCl_Three) δ: 156.4, 140.9, 128.2, 127.3, 126.1, 75.6, 65.1, 58.1, 55.2, 53.3, 31.7, 29.1, 28.9, 25.7, 23.6, 22.6, 14.0
[0050]
Preparation Example 15 Synthesis of (1R, 2R) -2-decylamino-3-pyrrolidino-1-phenyl-1-propanol
(1R, 2R) -2-decanoylamino-3-pyrrolidino-1-phenyl-1-propanol (181.8 mg, 0.486 mmol) was dissolved in methylene chloride (5 ml), and lithium aluminum hydride (153) was added at room temperature. 0.0 mg, 4.032 mmol) was added, and the mixture was refluxed at 35-40 ° C. for 2.5 hours. Then, 1N-hydrochloric acid (15 ml) was added on an ice bath, stirred for 30 minutes, and then saturated sodium bicarbonate solution (70 ml). Chloroform (100 ml) was added, and the organic layer was washed with 70 ml of water and saturated brine successively, dried over sodium sulfate, filtered, and the solvent was distilled off under reduced pressure. The obtained crude product was purified by silica gel column chromatography (elution solvent; chloroform: methanol = 20: 1, ethyl acetate: methanol = 2: 1), and the title material (121.2 mg, yield 69.) was obtained as a colorless oil. 3%).
TLC Rf 0.39 (CHCl_Three: MeOH = 9: 1), 0.19 (AcOEt: MeOH = 2: 1)
¹H-NMR (CDCl_Three) δ: 7.37-7.22 (5H, m, aromatic), 4.68 (1H, d, J = 3.90Hz, H-1), 2.99 (1H, m, H-2), 2.63-2.42 (8H, m, CH₂N (CH₂)₂, NHCH ₂), 1.77 (4H, m, H-3 ', H-4'), 1.41-1.24 (16H, m, (CH ₂)₈CH_Three), 0.88 (3H, t, CH_Three)
¹³C-NMR (CDCl_Three) δ: 143.1, 128.1, 127.0, 126.2, 73.9, 61.2, 57.6, 54.5, 48.5, 31.9, 30.2, 29.7, 29.6, 29.4, 29.3, 27.1, 23.6, 22.7, 14.1
[0051]
Preparation Example 16 Synthesis of (1R, 2R) -2-hexanoylamino-3-pyrrolidino-1-phenyl-1-propanol
(1R, 2R) -2-Amino-3-pyrrolidino-1-phenyl-1-propanol (160.0 mg, 0.727 mmol) was dissolved in methanol (4 ml), and triethylamine (0.121 ml, 0.873 mmol) at room temperature. ) Was added, n-hexanoyl chloride (0.102 ml, 0.727 mmol) was added on an ice bath, and the mixture was stirred at room temperature. Two hours later, triethylamine (0.121 ml, 0.873 mmol) and n-hexanoyl chloride (0.05 ml, 0.358 mmol) were added, and the mixture was stirred at room temperature. After 90 minutes, methanol (5 ml) was added, the solvent was distilled off under reduced pressure, ethyl acetate (40 ml) was added, and the organic layer was washed successively with saturated sodium hydrogen carbonate solution, water and saturated brine 25 ml each, and sodium sulfate. After drying above, it filtered and the solvent was depressurizingly distilled. The resulting crude product was purified by silica gel column chromatography (elution solvent; ethyl acetate: methanol = 2: 1) to obtain the title material (21.7 mg, yield 9.4%) as a colorless oil.
TLC Rf 0.12 (CHCl_Three: MeOH = 9: 1), 0.23 (AcOEt: MeOH = 2: 1)
¹H-NMR (CDCl_Three) δ: 7.36-7.23 (5H, m, aromatic), 5.86 (1H, d, J = 7.33Hz, NH), 5.04 (1H, d, J = 2.93Hz, H-1), 4.27-4.22 (1H, m, H-2), 2.85-2.83 (2H, m), 2.71-2.63 (4H, m), 2.09-2.04 (2H, m, COCH₂), 1.79 (4H, m), 1.52-1.44 (2H, m, COCH₂CH ₂), 1.29-1.20 (2H, m), 1.17-1.11 (2H, m), 0.84 (3H, t, CH_Three)
¹³C-NMR (CDCl_Three) δ: 173.4, 141.1, 128.2, 127.4, 125.9, 75.8, 58.1, 55.3, 52.4, 36.7, 31.2, 25.3, 23.7, 22.3, 13.8
[0052]
Preparation Example 17 (1R, 2R, 2 ′S) -2- (2′-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol and (1R, 2R, 2′R) -2- ( Synthesis of 2'-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol
(1R, 2R) -2-amino-3-pyrrolidino-1-phenyl-1-propanol (179.0 mg, 0.814 mmol) was dissolved in methylene chloride (6 ml), and 2-hydroxyoctanoic acid (166. 3 mg, 1.083 mmol) and N-hydroxysuccinimide (198.2 mg, 1.722 mmol) were added and stirred for 20 minutes, and then 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (171 on an ice bath. 0.1 mg, 0.893 mmol) was added, and the mixture was stirred overnight at room temperature. After confirming that the reaction was almost completed by TLC (chloroform: methanol = 7: 3), a saturated sodium hydrogen carbonate solution (20 ml) was added, followed by extraction with chloroform (30 ml × 3). The organic layer was dried over sodium sulfate and filtered, and the solvent was distilled off under reduced pressure. The resulting crude product was purified by silica gel column chromatography (elution solvent; chloroform: methanol = 20: 1, ethyl acetate: methanol = 2: 1), and the title material ((1R, 2R, 2 ′S) was obtained as a colorless oil. ) 6.9 mg and (1R, 2R, 2′R) 8.8 mg).
(1R, 2R, 2'S) -2- (2'-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol
TLC Rf 0.22 (CHCl_Three: MeOH = 9: 1), 0.32 (AcOEt: MeOH = 2: 1)
¹H-NMR (CDCl_Three) δ: 7.34-7.23 (5H, m, aromatic), 6.72 (1H, d, NH), 5.08 (1H, d, J = 2.92Hz, H-1), 4.24 (1H, m, H-2), 3.95 (1H, m, H-2 '), 2.98-2.85 (2H, m), 2.73-2.65 (4H, m), 1.80 (4H, m), 1.64 (1H, m, H-3'A), 1.42 (1H, m, H-3'B), 1.28-1.21 (8H, m, (CH ₂)_FourCH_Three), 0.87 (3H, t, CH_Three)
¹³C-NMR (CDCl_Three) δ: 174.1, 141.0, 128.3, 127.4, 125.8, 75.6, 71.9, 58.2, 55.4, 52.5, 34.8, 31.6, 29.0, 24.7, 23.7, 22.6, 14.0
[0053]
Preparation Example 18 Synthesis of (1R, 2R, 2'R) -2- (2'-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol
According to the method of Preparation Example 17, (2R) -2-hydroxyoctanoic acid was used instead of 2-hydroxyoctanoic acid to synthesize the title compound, and at the same time, the compound synthesized and separated in Preparation Example 17 was identified.
(1R, 2R, 2'R) -2- (2'-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol
TLC Rf 0.32 (CHCl_Three: MeOH = 4: 1), 0.20 (AcOEt: MeOH = 2: 1)
¹H-NMR (CDCl_Three) δ: 7.33-7.23 (5H, m, aromatic), 6.97 (1H, d, NH), 5.02 (1H, d, J = 3.41Hz, H-1), 4.35 (1H, m, H-2), 4.03 (1H, m, H-2 '), 3.09-3.04 (1H, m), 2.87-2.78 (5H, m), 1.85 (4H, brs), 1.58 (1H, m, H-3'A), 1.44 (1H, m, H-3'B), 1.27-1.21 (8H, m, (CH ₂)_FourCH_Three), 0.87 (3H, t, CH_Three)
¹³C-NMR (CDCl_Three) δ: 174.4, 140.6, 128.3, 127.6, 125.9, 75.1, 72.3, 57.9, 55.2, 51.7, 34.4, 31.6, 29.0, 24.6, 23.5, 22.6, 14.0
[0054]
Preparation Example 19 Synthesis of (1R, 2R) -2-benzyloxycarbonylamino-3-pyrrolidino-1-phenyl-1-propyl acetate
(1R, 2R) -2-Benzyloxycarbonylamino-1-phenyl-3-pyrrolidino-1-propanol 284.2 mg (0.803 mmol) was dissolved in 5 ml of pyridine, and 151.4 μl of acetic anhydride ( 1.61 mmol) was added, and the mixture was stirred at room temperature. After 16 hours, 38.0 μl (0.403 mmol) of acetic anhydride was added. After completion of the reaction, 30 ml of 1N hydrochloric acid was added and stirred for 20 minutes, followed by extraction with 50 ml of ethyl acetate, and the organic layer was washed with 30 ml of water, saturated sodium hydrogen carbonate solution, water and saturated brine successively, and over sodium sulfate. After drying, the mixture was filtered and the solvent was distilled off under reduced pressure. The combined washings were extracted three times with 70 ml of chloroform, the organic layer was dried over sodium sulfate and filtered, and the solvent was distilled off under reduced pressure. The obtained crude products were combined and purified by silica gel column chromatography (chloroform: methanol = 20: 1, ethyl acetate: methanol = 20: 1) to give 230.9 mg (yield 72.6%) of the title material as a colorless oil. )
TLC Rf. 0.24 (CHCl_Three: MeOH = 20: 1), 0.31 (AcOEt: MeOH = 9: 1)
¹H-NMR (CDCl_Three): 7.34-7.25 (10H, m, aromatic), 5.95 (1H, d, J = 4.88Hz, H-1), 5.10-4.94 (2H, m, COOCH₂), 4.17 (1H, m, H-2), 2.51 (6H, m, CH₂N (CH₂)₂), 2.03 (3H, s, COCH_Three), 1.73 (4H, m, H-3 ', H-4')
¹³C-NMR (CDCl_Three): 169.7, 156.1, 137.7, 136.6, 128.6, 128.3, 128.2, 128.0, 127.9, 126.6, 75.3, 66.5, 56.5, 54.5, 54.3, 23.5, 20.8
[0055]
Example 1
The inhibitory effect and cytotoxicity of the compounds of the present invention and D-threo-PDMP on glucosylceramide synthase were compared.
[Method and results]
The inhibitory activity against glucosylceramide synthase was measured by the method reported by Inoguchi and Radin (J. Lipid Res. Vol.28, 565-571, 1987) using the microsomal fraction of mouse liver as the enzyme source. Of 50% enzyme activity against glucosylceramide synthase (IC)₅₀). In addition, the cytotoxicity of each compound on cultured human fetal fibroblasts was cultured for 24 hours, and then 50% by the MTT method (Pauwels et al, J. Virol. Methods, Vol. 20, pp.309-321, (1988)). % Concentration of cells killed (CD₅₀). Furthermore, if the concentration showing cytotoxicity is high and the concentration inhibiting glucosylceramide synthase is low, it can be said that the safety of the compound is high.₅₀/ IC₅₀The value was calculated. These results are shown in Table 1. As a result, the compound of the present invention had significantly higher inhibitory activity against glucosylceramide synthase than D-threo-PDMP, and no cytotoxicity was observed in the concentration range examined. CD₅₀/ IC₅₀In terms of value, D-threo-PDMP was 5, whereas all of the compounds of the present invention were high, and it was found that they were excellent in safety.
[0056]
[Table 1]

[0057]
Example 2
The therapeutic effect of the compound of the present invention of Preparation Example 3 on human esophageal cancer was examined.
[Method and results]
The cell line KES-3 established from human esophageal cancer is 1 × 10⁷The mice were transferred subcutaneously to the dorsal region of nude mice, and 10 μl of a physiological saline solution (7.78 mg / ml) of the hydrochloride of the compound of Preparation Example 3 once a day for 7 days at the time of tumor formation after 1 to 2 weeks. Local injection and histopathological changes were examined. As a comparative control, the same amount of the physiological saline solution was locally administered. The results are shown by optical micrographs 1 and 2 (magnification 240).
When comparing the transplanted tissue images (hematoxyeosin staining) of the above administration experiment of KES-3, the group of the compound of the present invention in Preparation Example 3 was generally compared with the tissue image of the physiological saline control group (Photo 1). In particular, nuclei were evacuated and condensed, and cell swelling was observed, and obvious degeneration was observed (Photo 2).
[0058]
【The invention's effect】
According to the present invention, a drug containing an amino alcohol derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, particularly exhibiting abnormal cell proliferation in an undifferentiated state like a malignant tumor. It is possible to provide a medicament effective for treating diseases based on the above, and a promising medicament as a solid cancer therapeutic agent.
[Brief description of the drawings]
FIG. 1 is an optical micrograph 1 showing histopathological changes when physiological saline is locally injected after transplantation of KES-3 into nude mice.
FIG. 2 is a photomicrograph 2 showing histopathological changes when a physiological saline solution of the hydrochloride of the compound of Preparation Example 3 is locally injected after transplantation of KES-3 into nude mice.

Claims (13)

A pharmaceutical comprising an amino alcohol derivative represented by the following general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Chemical 1]
R ² —CH ₂ —C * H (NHP ¹ ) —C * H (OR ³ ) —R ¹ (I)
(Wherein * represents an asymmetric carbon, R ¹ represents a phenyl group, R ² represents a pyrrolidino, piperidino or piperazino group optionally substituted with a hydroxyl, methyl or hydroxymethyl group, or a morpholino group, R ³ represents a hydrogen atom, P ¹ represents an alkyl group having 4 to 16 carbon atoms, aralkyloxycarbonyl group having 6 to 18 carbon atoms, an alkoxycarbonyl group having 4 to 16 carbon atoms or alkenyl having 4 to 16 carbon atoms, Represents an oxycarbonyl group .) In the general formula (I), R ¹ is a phenyl group, R ² is a pyrrolidino, hydroxypyrrolidino, piperidino, hydroxypiperidino or morpholino group, R ³ is a hydrogen atom, and P ¹ has 4 carbon atoms. The medicament according to claim 1, which is an alkyl group of -16, an aralkyloxycarbonyl group having 6-18 carbon atoms, an alkoxycarbonyl group having 4-16 carbon atoms, or an alkenyloxycarbonyl group having 4-16 carbon atoms. A therapeutic agent for an abnormal proliferative disease comprising an amino alcohol derivative represented by the following general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Chemical 2]
R ² —CH ₂ —C * H (NHP ¹ ) —C * H (OR ₃ ) —R ¹ (I)
(Wherein * represents an asymmetric carbon, R ¹ represents a phenyl group, R ² represents a pyrrolidino, piperidino or piperazino group optionally substituted with a hydroxyl, methyl or hydroxymethyl group, or a morpholino group, R ³ represents a hydrogen atom, P ¹ represents an alkyl group having 4 to 16 carbon atoms, aralkyloxycarbonyl group having 6 to 18 carbon atoms, an alkoxycarbonyl group having 4 to 16 carbon atoms or alkenyl having 4 to 16 carbon atoms, Represents an oxycarbonyl group .) In the general formula (I), R ¹ is a phenyl group, R ² is a pyrrolidino, hydroxypyrrolidino, piperidino, hydroxypiperidino or morpholino group, R ³ is a hydrogen atom, and P ¹ has 4 carbon atoms. The abnormal proliferative disease according to claim 3 , which is an alkyl group having 16 to 16 carbon atoms, an aralkyloxycarbonyl group having 6 to 18 carbon atoms, an alkoxycarbonyl group having 4 to 16 carbon atoms, or an alkenyloxycarbonyl group having 4 to 16 carbon atoms. Remedy. The treatment for an abnormal proliferative disease according to claim 3 , wherein, in the general formula (I), R ¹ is a phenyl group, R ² is a pyrrolidino group, R ³ is a hydrogen atom , and P ¹ is a benzyloxycarbonyl group. medicine. The therapeutic agent for an abnormal proliferative disease according to any one of claims 3 to 5 , wherein in the general formula (I), the configuration is (1R, 2R). The therapeutic agent according to any one of claims 3 to 6 , wherein the abnormal proliferative disease is cancer. (1R, 2R) -2-Hexanoylamino-3-pyrrolidino-1-phenyl-1-propanol or a pharmaceutically acceptable salt thereof. (1R, 2R, 2'R) -2- (2'-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol or a pharmaceutically acceptable salt thereof. The (1R, 2R) -2-hexanoylamino-3-pyrrolidino-1- according to claim 8. A medicament comprising phenyl-1-propanol or a pharmaceutically acceptable salt thereof as an active ingredient. The (1R, 2R, 2′R) -2- (2′-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol or a pharmaceutically acceptable salt thereof according to claim 9 is effective. A pharmaceutical containing as an ingredient. A therapeutic agent for an abnormal proliferative disease comprising (1R, 2R) -2-hexanoylamino-3-pyrrolidino-1-phenyl-1-propanol or a pharmaceutically acceptable salt thereof according to claim 8 as an active ingredient. The (1R, 2R, 2′R) -2- (2′-hydroxyoctanoylamino) -3-pyrrolidino-1-phenyl-1-propanol or a pharmaceutically acceptable salt thereof according to claim 9 is effective. A therapeutic agent for abnormal proliferative diseases, which is included as an ingredient

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