JP4152576B2 - Anti-cancer agent - Google Patents

Description

〔式中、Ｒは
【化５】

又は
【化６】

を表わす〕で示される化合物若しくはその塩が，シスプラチンに比して毒性が少なく、しかも、シスプラチンと比較して、
種々の癌に対し大きい制がん作用を有すること及び線維芽細胞の増殖を抑制することを発見して本発明に至った。
【０００６】
即ち、本発明は、一般式〔Ｉ〕
【化７】

〔式中、Ｒは
【化８】

又は
【化９】

を表す〕で示される化合物若しくはその塩を含有することを特徴とする抗がん剤にある。
本発明における、一般式〔Ｉ〕
【化１０】

〔式中、Ｒは
【化１１】

又は
【化１２】

を表す〕で示される化合物（以下、前記一般式〔Ｉ〕で示される化合物という）若しくはその塩（以下、前記一般式〔Ｉ〕で示される化合物等という）及びそれらの製造方法は、国際公開番号ＷＯ９２／００９６２号の国際公開公報及び特開平４−６９３７７号公報（以下、前記公報等という）に開示されている。前記公報等には、さらに、前記一般式〔Ｉ〕で示される化合物等が、心抑制作用を伴うことなく、心筋の過収縮及び過伸展を抑制して、心筋を壊死から保護する作用を有すること、心筋梗塞の治療及び予防効果を有すること、並びに心筋壊死の抑制及び予防効果を有することなどが開示されている。
【０００８】
本発明者らは、生体内（in vivo）及び生体外（in vitro）において、前記一般式〔Ｉ〕で示される化合物等が、シスプラチン及びその他抗がん剤に比して、種々のがんに対して優れた抗がん作用を有し、種々の癌に対する広い抗がんスペクトルを示すことを発見し、また、前記一般式〔Ｉ〕で示される化合物等が、線維芽細胞の増殖を抑制することを発見した。
【０００９】
本発明において、抗がん剤の用語は、がんの治療剤及び／又は線維化抑制剤を包含する。
本発明において、前記一般式〔Ｉ〕で示される化合物の「塩」は、医薬的に許容されうる塩を意味し、例えば、塩酸塩、臭化水素酸塩、硫酸塩、リン酸塩又は硝酸塩等の無機酸付加塩；酢酸塩、プロピオン酸塩、コハク酸塩、グリコール酸塩、乳酸塩、リンゴ酸塩、シュウ酸塩、酒石酸塩、クェン酸塩、マレイン酸塩、フマール酸塩、メタンスルホン酸塩、ベンゼンスルホン酸塩、ｐ−トルエンスルホン酸塩又はアスコルビン酸塩等の有機酸付加塩、或いはアスパラギン酸塩又はグルタミン酸塩等のアミノ酸付加塩を含み、さらに含水物及び水和物を含むが、これらに限定されるものではない。
【００１０】
本発明に係る前記一般式〔Ｉ〕で示される化合物等は、優れた制がん作用を有する。すなわち、本発明の抗がん剤は、前記一般式〔Ｉ〕で示される化合物等を主薬として含有する。
本発明の抗がん剤は、他の抗がん剤と併用投与することにより、耐性化したがんに対しても抗がん剤の制がん作用を発揮させることができる。本発明に係る前記一般式〔Ｉ〕で示される化合物等を抗がん剤として用いる場合、通常、全身的又は局所的に、或いは経口又は非経口で投与することができる。本発明の抗がん剤は、他の抗がん剤と同時に投与してもよく、又は他の抗がん剤の投与前、又は投与後に投与してもよい。
【００１１】
投与量は、年齢、体重、症状、治療効果、投与方法及び処理時間等により異なるが、通常成人（平均体重６０ｋｇ）一人当たり、０．０１ｍｇ乃至１ｇの範囲内で、好ましくは、１００乃至５００ｍｇの範囲内で、一日一回若しくは一日数回に分けて経口又は非経口で投与される。非経口で投与する場合には、１２時間以上に亘って持続的に投与することができる。
【００１２】
本発明の前記一般式〔Ｉ〕で示される化合物等を主薬とし、経口投与のための固体組成物を調製する場合には、錠剤、丸剤、散剤、顆粒剤等の剤形が可能である。このような固体組成物においては、一種又は二種以上の主薬を、一以上の活性な希釈剤、分散剤又は吸着剤等、例えば乳糖、マンニトール、ブドウ糖、ヒドロキシプロピルセルロース、微晶性セルロース、澱粉、ポリビニルビロリドン、メタケイ酸アルミン酸マグネシウム又は無水ケイ酸末等と混合することができる。又、組成物は常法に従って、希釈剤以外の添加剤を混合させてもよい。
【００１３】
本発明の前記一般式〔Ｉ〕で示される化合物等を主薬として、錠剤又は丸剤に調製する場合には、必要により、白糖、ゼラチン、ヒドロキシプロピルセルロース又はヒドロキシメチルセルロースフタレート等の胃溶性あるいは腸溶性物質のフィルムで被覆してもよいし、二以上の層で被覆してもよい。さらに、ゼラチン又はエチルセルロースのような物質によりカプセルにしてもよい。
【００１４】
本発明の前記一般式〔Ｉ〕で示される化合物等を主薬として、経口投与のための液体組成物を調製する場合には、薬剤的に許容される乳濁剤、溶解剤、懸濁剤、シロップ剤又はエリキシル剤等の剤形が可能である。この場合、用いられる希釈剤としては、例えば精製水・エタノール、植物油又は乳化剤等がある。また、これらの組成物には、希釈剤以外に浸潤剤、懸濁剤、甘味剤、風味剤、芳香剤又は防腐剤等のような補助剤を混合させてもよい。
【００１５】
本発明の前記一般式〔Ｉ〕で示される化合物等を主薬として、非経口投与のための注射剤に調製する場合は、無菌の水性若しくは非水性の溶液剤、可溶化剤、懸濁剤又は乳化剤を用いることができる。水性の溶液剤、可溶化剤、懸濁剤としては、例えば注射用水、注射用蒸留水、生理食塩水、シクロデキストリン及びその誘導体、トリエタノールアミン、ジエタノールアミン、モノエタノールアミン、トリエチルアミン等の有機アミン類或いは無機アルカリ溶液等がある。
【００１６】
本発明の前記一般式〔Ｉ〕で示される化合物等を主薬として、水溶性の溶液剤にする場合、例えば、プロピレングリコール、ポリエチレングリコール若しくはオリーブ油のような植物油、又はエタノールのようなアルコール類等を用いてもよい。また、可溶化剤としては、例えば、ポリオキシエチレン硬化ヒマシ油、蔗糖脂肪酸エステル等の界面活性剤（混合ミセル形成）、或いはレシチン又は水添レシチン（リポソーム形成）等を使用することができる。さらに、植物油等非水溶性の溶解剤と、レシチン、ポリオキシエチレン硬化ヒマシ油又はポリオキシエチレンポリオキシプロピレングリコール等とからなるエマルジョン製剤にすることもできる。
非経口投与のためのその他の組成物としては、一種又は二種以上の主薬、即ち、前記一般式〔Ｉ〕で示される化合物等を含み、それ自体公知の方法により処方される外用液剤、軟膏のような塗布剤、座剤又はペッサリー等にしてもよい。
【００１７】
実施例
以下に、本発明の前記一般式〔Ｉ〕で示される化合物等を抗がん剤の主薬とする製剤例について具体的に説明する。しかし、本発明は以下の説明により何ら限定されるものではない。
【００１８】
例１
本実施例の抗がん剤の注射製剤においては、主薬として、１−［１−４−（ジフェニルメチル）ピペラジニル］−３−［１−〔４−（４−クロロフェニル）−４−ヒドロキシ〕ピペリジニル］−２−プロパノール（以下、化合物１という）が用いられている。
前記化合物１の合成例について、以下に説明する。なお、以下の説明において、核磁気共鳴スペクトル（ＮＭＲ）の測定は、テトラメチルシランを内部標準として行ってｐｐｍにて表示してある。部は容量部を示す。
【００１９】
(１) １−（ジフェニルメチル）−４−（１−（２，３−エポキシ）プロピル）ピペラジンの合成
１−（ジフェニルメチル）ピペラジン（10.0ｇ）をアセトニトリル（50ｍｌ）に溶かし、炭酸ナトリウム（6.5ｇ）及びエピブロモヒドリン（6．8ｇ）を加え、２．５時間加熱還流した。塩を濾別後、濾液を減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー（ワコーゲルＣ−200、200ｇ）にて精製し、クロロホルム99部とメタノール１部の混合溶媒により溶出し、１−（ジフェニルメチル）−４−（１−（２，３−エポキシ）プロピル）ピペラジン（5.9ｇ）を得た。
核磁気共鳴スペクトル
¹Ｈ−ＮＭＲ（ＣＤＣ１3、５00ＭＨｚ）δ：2.30〜2.80（12Ｈ，ｍ）、3.06〜3.10（1Ｈ，ｍ）4.23（１Ｈ，ｓ）、7.16（2Ｈ，ｔ，J＝7.3Ｈｚ）、7.25（4Ｈ．ｔ，Ｊ＝7.3Ｈｚ）、7.40（4Ｈ，ｄ，Ｊ＝7.3Ｈｚ）。
【００２０】
(２) １−〔１−４−（ジフェニルメチル）ピペラジニル〕−３−［１−〔４−（４−クロロフェニル）−４−ヒドロキシ〕ピペリジニル］−２−プロパノールの合成
１−（ジフェニルメチル）−４−（１−（２，３−エポキシ）プロピル）ピペラジン（3.0ｇ）および４−（４−クロロフェニル）−４−ヒドロキシピペリジン（2.5 ｇ）をｏ−ジクロロベンゼン（20ｍｌ）に溶解し、２．５時間加熱還流した。放冷後シリカゲルカラムクロマトグラフィー（ワコーゲルＣ−200、100ｇ）にて精製し、１−〔１−４−（ジフェニルメチル）ピペラジニル〕−３−［１−〔４−（４−クロロフェニル）−４−ヒドロキシ〕ピペリジニル］−２−プロパノール（化合物１)の4.6ｇを得た。
赤外吸収スペクトル：ＩＲ ν ｍａｘ（ｃｍ-1）ＫＢｒ：3300，2950，2650，1620，1450，1100，910，830，750，710（塩酸塩として）
核磁気共鳴スペクトル
¹Ｈ−ＮＭＲ（ＣＤＣ１3、500 ＭＨｚ）δ：1.50〜1.90（４Ｈ，ｍ）、2.01〜2.21（２Ｈ，ｍ）2.30〜2.55（１０Ｈ，ｍ）、2.80〜2.90
（２Ｈ，ｍ）、3.87〜3.93（１Ｈ，ｍ）、4.22（１Ｈ，ｓ）、7.16（２Ｈ，ｔ，Ｊ＝7.3Ｈｚ）、7.26（４Ｈ．ｔ，Ｊ＝7.3Ｈｚ）、7.30（２Ｈ，ｄ，Ｊ＝8.5 Ｈｚ）、7.40（４Ｈ，ｄ，Ｊ＝7.3 Ｈｚ）、7.42（２Ｈ，ｄ，Ｊ＝8.5 Ｈｚ）。
ＦＤマススペクトル
ＦＤ−ＭＳ（ｍ／ｚ）：519，521（Ｍ+）。
【００２１】
（１）化合物１の注射製剤
化合物１ ２〜４０ｍｇ
Ｄ−ソルビトール １０００ｍｇ
クエン酸 １０ｍｇ
水酸化ナトリウム 適量
注射用水 注射用水を加えて２０．０ｍ１とする。
Ｄ−ソルビトール及びクエン酸を充分量の注射用水に溶解した。得られた溶液に化合物１を溶解させ、水酸化ナトリウムで、ｐＨを３．２〜３．３に調節した。次いで、攪拌しつつ、残りの注射用水を加えた。この溶液を濾過し、２０．０ｍ１アンプルに詰めて封入した。このアンプルの内容物をオートクレーブにかけて滅菌した。
【００２２】
（２）各種ヒトがん細胞株における in vitro 抗腫瘍効果についてのＭＴＴアッセイ法による検討。
ヒト非小細胞肺臓癌細胞株ＰＣ−１４、ヒト小細胞肺臓癌細胞株ＳＢＣ−３、ヒト乳癌細胞株ＭＣＦ−７、ヒト卵巣癌細胞株ＳＫＯＶ３、ヒト白血病細胞株ＨＬ６０及びヒト大腸癌細胞株ＷｉＤＲを、夫々、ＲＰＭＩ
１６４０培地中でトリプシン処理又はセルスクレイパーを用いてシングル細胞とし、１５μｌ当たり１００個の細胞浮遊液を調製した。これに抗がん剤として化合物１をジメチルスルホキシドに溶解させて、化合物１の濃度が０．５乃至１μＭの範囲となるように加え、これを９６穴プレートに１ウェル当たり１５０μｌづつ入れた。このプレートを、３７℃の温度下で、５％の二酸化炭素濃度及び飽和水蒸気の条件下に、９６時間保持し、培養した。培養後、Ｄ−ＰＢＳ(-)に５ｍｇ／ｍ１の濃度で溶解させたＭＴＴ〔３−（４，５−ジメチルチアゾール−２−イル）−２，５−ジフェニルテトラゾリウムブロマイド（3-（4,5-dimethy1thiazo1-2-y1）-2,5-diphenyltetrazoliumbromide）〕試薬を、２０μｌ加えて、３７℃で更に４時間培養した。培養終了後、プレートごと遠心し、上清を捨てた。２００μｌのジメチルスルホキシドを加えて、黄色のＭＴＴが、がん細胞内のミトコンドリアにある脱水素酵素の作用により生成した紫色のホルマザンを溶解させ、生成したホルマザンの量を、波長５６２〜６３０ｎｍについての吸光度をマルチプレートリーダーにより測定した。陰性対照の平均発育率を０％、陽性対照の平均発育率を１００％として腫瘍容量発育曲線を描き、化合物1の、５０％のヒトがん細胞増殖抑制濃度を算出した。これを表１に示す。
【００２３】
表１

表１の値は、化合物１の使用下で３回の独立したがん細胞の培養実験におけるＭＴＴアッセイの５０パーセントのがん細胞増殖抑制濃度（ＩＣ₅₀）値を平均値±誤差範囲（μＭ）で示す。
【００２４】
表１に示すように、化合物１は、０．５乃至１．１μＭの濃度で、各種がんについての５０％増殖抑制効果（ＩＣ₅₀）を示した。この化合物１についてのＩＣ₅₀値は、固形癌に対し最も有効であるとされているシスプラチンにおけるＭＴＴアッセイ法によるＩＣ₅₀値の２乃至３μＭに比して、遥かに低い値であり、化合物１のＩＣ₅₀の値より、化合物１の制がん効果が大きいことが分かり、抗がん剤として有効である。また、肺臓癌、乳癌、大腸癌、卵巣癌等の固形癌細胞に対しても、白血病細胞に対してと同程度の抗腫瘍効果を示すことは、広い抗腫瘍スペクトラムを有するものである。
【００２５】
例２
対照、化合物１の濃度が１×１０^{- ６}Ｍの系列及び化合物１の濃度が３×１０^-6Ｍの系列の夫々に、９枚のシャーレ（合計２７枚）を準備した。２７枚のシャーレの夫々に、シャーレ当たり４．４×１０⁵個の繊維芽細胞ＮＩＨ３Ｔ３を加え、夫々に、１０％ＦＢＳ(ウシ胎仔血清)を添加したＤ−ＭＥＭ培養液を加えた。対照には、化合物１の添加量相当の水を加えて培養した。２４時間毎に、各系列３枚の細胞数を測定し、平均して、シャーレ当たりの平均細胞数とした。その結果を図１に示す。この結果に示されるように、化合物１は，抗がん作用と共に、線維芽細胞の増殖による間質性肺炎やケロイドなどの増殖性病変に対しても有効である。
【００２６】
例３
雌性ヌードマウスＢＡＬＢ／ｃ ｎｕ／ｎｕ（６週齢）に、２×１０⁷個のヒト非小細胞肺臓がん株ＰＣ−１４の細胞を生理食塩水浮遊液とし、背部皮下に植え込んで生着の経過を観察した。移植後７日目に腫瘍の状態を確認して試験可能な個体を選出し、偏りが生じないように各個体のランダマイズを行って、夫々６匹宛、対照系列、第一被検系列及び第二被検系列に分けた。各々の移植された腫瘍の腫瘍容積を、腫瘍の短径及び腫瘍の長径を測定して、腫瘍の短径の二乗と腫瘍長径の積、即ち
式:(腫瘍の短径)^２×(腫瘍の長径)により求めた。腫瘍の移植後、第７日目より化合物１の投与を開始した。
化合物１は、その１．２ｍｇを０．０５ｍｌのジメチルスルホキシドに溶解させ、０．９５ｍ１の５％ソルビトール−０．２％クエン酸１水和物溶液（ｐＨ３．３）を加えて均一な溶液とし、化合物１の注射液とした。
化合物１の投与は、系列番号１の被検体群の６匹には、腫瘍移植後の７日目、８日目、９日目、１０日目及び１１日目に、夫々、一日一回宛、体重１ｋｇ当たり３ｍｇの化合物１が、ヌードマウスの尻尾から注入され、系列番号２の被検体群の６匹には、腫瘍移植後の７日目、８日目、９日目、１０日目及び１１日目に、夫々、一日一回宛、体重１ｋｇ当たり５ｍｇの化合物１が、ヌードマウスの尻尾から注入された。対照となる系列番号１の被検体群は、化合物１が投与されない系列であり、化合物１の投与及び治療は一切行われなかった。総ての被検体群について、化合物１の投与第１日目から第８日目まで毎日一回、化合物１の投与する前の時点で、毎日一回、各マウスの全身状態についての観察並びに各マウスの体重及び腫瘍容積についての測定を行った。マウスの全身状態についての観察及び体重の測定結果においては、系列番号１乃至３の被検体群の間で殆ど差がみられなかったが、マウスの腫瘍容積については、系列番号１乃至３の被検体群の間で差がみられた。腫瘍容積についての測定結果を次の表２に示し、その系列番号毎の、被検体群の腫瘍容積の平均値を表３に示す。表２及び表３において、第２日目以降の腫瘍容積の大きさは、第１日目の腫瘍容積の大きさを１００として示されている。
【００２７】
表２

【００２８】
表３

１−〔１−４−（ジフェニルメチル）ピペラジニル〕−３−［１−〔４−（４−クロロフェニル）−４−ヒドロキシ〕ピペリジニル］−２−プロパノール
例１乃至例３の結果よりみて、化合物１による制がん効果が大きいことが分かる。
【００２９】
前記例１乃至３においては、化合物１の１−［１−４−（ジフェニルメチル）ピペラジニル］−３−［１−〔４−（４−クロロフェニル）−４−ヒドロキシ〕ピペリジニル］−２−プロパノールが、主薬として用いられているが、１−〔２−(１，２，３，４−テトラヒドロ）イソキノリニル〕−３−〔１−（４−ジフェニルメチル）ピペラジニル〕−２−プロパノールを主薬として用いても、同様の結果を得ることができた。
【００３０】
１−〔２−(１，２，３，４−テトラヒドロ）イソキノリニル〕−３−〔１−（４−ジフェニルメチル）ピペラジニル〕−２−プロパノールの合成例について、以下に説明する。なお、以下の説明において、核磁気共鳴スペクトル（ＮＭＲ）の測定は、テトラメチルシランを内部標準として行ってｐｐｍにて表示してある。部は容量部を示す。
【００３１】
(１) ２−〔１−（２，３−エポキシ）プロピル〕−１，２，３，４−テトラヒドロイソキノリンの合成
１，２，３，４−テトラヒドロイソキノリン（25.0 ｇ）をアセトニトリル（100 ｍｌ）に溶解し、これに炭酸ナトリウム（40.0 ｇ）およびエピブロモヒドリン（31.0 ｇ）を加え４時間加熱還流した。塩を濾別後濾液を減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー（ワコーゲルＣ−200、500ｇ）にて精製し、クロロホルム99部とメタノール１部の混合溶媒により溶出し、２−〔１−（２，３−エポキシ）プロピル〕−１，２，３，４−テトラヒドロイソキノリン（15.6
ｇ）を得た。
核磁気共鳴スペクトル
¹Ｈ−ＮＭＲ（ＣＤＣ１3、100 ＭＨｚ）δ：2.36〜2.60（２Ｈ，ｍ）、2.73〜3.03（６Ｈ，ｍ）、3.09〜3.29（１Ｈ，ｍ）、3.65（１Ｈ，ｄ，Ｊ＝14.9Ｈｚ）、3.83（１Ｈ，ｄ，
【００３２】
例４
１−〔２−（１，２，３，４−テトラヒドロ）イソキノリニル〕−３−〔１−（４−ジフェニルメチル）ピペラジニル〕−２−プロパノールの合成
２−［１−（２，３−エポキシ）プロピル）−１，２，３，４−テトラヒドイソキノリン（3.0ｇ）および１−（ジフェニルメチル）ピペラジン（4.4ｇ）を、ｏ−ジクロロベンゼン（20ｍｌ）に溶解し、２．５時間の間加熱還流した。放冷後シリカゲルカラムクロマトグラフィー（ワコーゲルＣ−200、150ｇ）にて精製し、１−〔２−（１，２，３，４−テトラヒドロ）イソキノリニル〕−３−〔１−（４−ジフェニルメチル）ピペラジニル〕−２−プロパノール（ 6.0ｇ）を得た。
赤外吸収スペクトル：ＩＲ νｍａｘ（ｃｍ-1）ＫＢｒ：3400，3000，2550，1620，1450，1080，920，760，710（塩酸塩として）
核磁気共鳴スペクトル
¹Ｈ−ＮＭＲ（ＣＤＣ１3、100ＭＨｚ）δ：2.30〜2.60（1,2Ｈ，ｍ）、2.75〜2.95（４Ｈ，ｍ）3.62〜3.80（２Ｈ，ｍ）、3.92〜4.03（１Ｈ，ｍ）、4.21（１Ｈ，ｓ）、7.00〜7,51（14Ｈ，ｍ）。
ＦＤマススペクトル
ＦＤ−ＭＳ（ｍ／ｚ）：441（Ｍ+）。
【００３３】
【発明の効果】
本発明に係る前記一般式〔Ｉ〕で示される化合物等は、従来の抗がん剤、例えばシスプラチンに比して毒性が少なく、しかも、従来の抗がん剤に比して、種々のがんに対し大きい制がん作用を有し、広い制がんスペクトルを示すものであり、従来の抗がん剤に比して、がん(固形癌)及びその他の悪性腫瘍等のがん治療の上で、大きく貢献するものである。また本発明は、肺線維症等の、線維芽細胞の増殖を抑制する作用を有し、従来の肺線維症やケロイドなどの治療薬に比して、優れた線維化抑制作用を有し、肺線維症やケロイド治療の上で大きく貢献するものである。
【図面の簡単な説明】
【図１】 例２に示す化合物１の繊維芽細胞に対する細胞増殖抑制作用を示すグラフである。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an anticancer agent that suppresses the growth of cancer cells, that is, an antineoplastic agent, and in particular, an anticancer agent containing a diphenylmethylpiperazine derivative that suppresses the growth of cancer cells of mammals including humans. The present invention relates to cancer drugs, that is, anticancer drugs. The present invention also relates to a pulmonary fibrosis disease caused by fibroblast proliferation and a fibrosis inhibitor for treating proliferative lesions of pulmonary fibrosis disease caused by fibroblast proliferation as a side effect of an anticancer agent, In particular, the present invention relates to a fibrosis inhibitor for treating proliferative lesions in mammals including humans, which contains a diphenylmethylpiperazine derivative.
[0002]
[Prior art]
Anticancer agents or antineoplastic agents include (1) nitrogen mustards such as melphalan, mechlorethamine and cyclophosphamide, (2) nitrosoureas such as BCNU, CCNU, methyl-CCNU and ACNU, ( 3) Azirine and epoxides such as thioteba, mitomycin AZQ, carbocon, dianhydrogalactitol and dibromodalcitol, (4) alkylating agents such as procarbazine, dacarbazine and hexamethylenemelamine, (5) metrexate (MTX), 6 -Metabolism of 5-fluoropyrimidine and related compounds such as mercaptopurine (6-MP), 6-thioguanine (6-TG) and 5-fluorouracil (5-FU), tegafur, UFT, 5'-DFUR and HCFU Antagonists, (6) pinklistin, pin plastin and Plant-derived anticancer agents such as pinca alkaloid compounds such as pindesine, etoposide compounds such as etoposide and teniposide, taxanes such as baclitaxel and docetaxel, and camptothecin compounds such as irinotecan, (7) adriamycin, servidin, actinomycin D Anticancer antibiotics such as cosmegen, prenoxane, mutamycin, medamicin and novantrone, (8) adenocorticoids, estrogens, progestins, antiestrogens, aromatase inhibitors, hormones such as androgens, antiandrogens and LH-RH analogs, (9) Enzymes such as L-asparaginase, (10) Platinum complex compounds such as cisplatin, carboplatin and nedaplatin (254-S), (11) non-specific immunostimulatory substances, (12) interferon and (13) TNF And the like.
[0003]
[Problems to be solved by the invention]
As treatments with these anticancer agents are established, for example, acute lymphocytic leukemia and Hodgkin's disease can return to the social life of patients with considerable frequency. However, for solid cancers such as gastric cancer, lung cancer and colon cancer, most depend on surgical therapy and radiation therapy. Cisplatin is used for the treatment of solid cancer because cisplatin has a broad antitumor spectrum for these solid cancers. However, nephrotoxicity, gastrointestinal toxicity, auditory toxicity and peripheral neurotoxicity have been observed for cisplatin, and in addition, the cure rate is considered to be low.
Moreover, it is known that some anticancer agents have pulmonary fibrosis as a side effect and have a significant effect on the prognosis of life.
The object of the present invention is to solve the problems of these anticancer agents.
[0004]
[Means for Solving the Problems]
The present invention has a large anticancer activity against various cancers (that is, an anti-malignant tumor activity) or a property of suppressing the proliferation of lung fibroblasts, or has a large resistance against various cancers. An object of the present invention is to provide a compound having low toxicity and a salt thereof and a derivative thereof, which have both cancer activity (that is, anti-malignant tumor activity) and a property of suppressing proliferation of lung fibroblasts.
[0005]
The inventors of the present invention have the general formula [I]
[Formula 4]

[Where R is embedded image

Or [Chemical formula 6]

Or a salt thereof is less toxic than cisplatin, and compared with cisplatin,
The present inventors have found that it has a large anticancer effect against various cancers and suppresses the proliferation of fibroblasts.
[0006]
That is, the present invention relates to the general formula [I]
[Chemical 7]

[Where R is embedded image

Or [Chemical 9]

It is an anticancer agent characterized by containing a compound represented by
In the present invention, the general formula [I]
[Chemical Formula 10]

[Wherein R is embedded image

Or [Chemical Formula 12]

Compounds represented by the representative] (hereinafter, the referred compounds of the formula represented by (I)) or a salt thereof (hereinafter, the general formula [that compounds represented by I]) and methods for their preparation are described in International Publication This is disclosed in International Publication No. WO92 / 00962 and Japanese Patent Laid-Open No. 4-69377 (hereinafter referred to as the above publication). In the above publications, the compound represented by the general formula [I] has an action of protecting the myocardium from necrosis by suppressing myocardial overcontraction and hyperextension without accompanying the cardiac inhibitory action. In other words, it has a therapeutic and preventive effect on myocardial infarction, and has an inhibitory and preventive effect on myocardial necrosis.
[0008]
The present inventors have found that the compounds represented by the above general formula [I] can be used in various cancers in vivo and in vitro compared to cisplatin and other anticancer agents. It has been found that it has an excellent anticancer activity against various cancers and exhibits a wide anticancer spectrum against various cancers, and the compound represented by the general formula [I], etc., promotes the proliferation of fibroblasts. Found to suppress.
[0009]
In the present invention, the term anticancer agent includes a therapeutic agent for cancer and / or a fibrosis inhibitor.
In the present invention, the “salt” of the compound represented by the general formula [I] means a pharmaceutically acceptable salt, for example, hydrochloride, hydrobromide, sulfate, phosphate or nitrate. Inorganic acid addition salts such as acetate, propionate, succinate, glycolate, lactate, malate, oxalate, tartrate, kenate, maleate, fumarate, methanesulfone Organic acid addition salts such as acid salts, benzenesulfonic acid salts, p-toluenesulfonic acid salts and ascorbic acid salts, or amino acid addition salts such as aspartic acid salts and glutamic acid salts, and further hydrates and hydrates. However, it is not limited to these.
[0010]
The compound represented by the general formula [I] according to the present invention has an excellent anticancer effect. That is, the anticancer agent of this invention contains the compound etc. which are shown by the said general formula [I] as a main ingredient.
The anticancer agent of the present invention can exert the antitumor action of the anticancer agent even against resistant cancer by being administered in combination with another anticancer agent. When the compound represented by the above general formula [I] according to the present invention is used as an anticancer agent, it can be generally administered systemically or locally, or orally or parenterally. The anticancer agent of the present invention may be administered simultaneously with other anticancer agents, or may be administered before or after administration of other anticancer agents.
[0011]
The dose varies depending on age, body weight, symptoms, therapeutic effect, administration method and treatment time, etc., but is usually within the range of 0.01 mg to 1 g, preferably 100 to 500 mg per adult (average weight 60 kg). Within the range, it is administered orally or parenterally once a day or divided into several times a day. When administered parenterally, it can be administered continuously over 12 hours.
[0012]
When preparing a solid composition for oral administration using the compound represented by the above general formula [I] of the present invention as the main drug, dosage forms such as tablets, pills, powders, granules and the like are possible. . In such a solid composition, one or more active ingredients are added to one or more active diluents, dispersants or adsorbents such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch. , Polyvinyl pyrrolidone, magnesium aluminate metasilicate, or anhydrous silicic acid powder. Further, the composition may be mixed with additives other than the diluent according to a conventional method.
[0013]
When preparing a tablet or pill using the compound represented by the above general formula [I] of the present invention as a main drug, if necessary, gastric or enteric properties such as sucrose, gelatin, hydroxypropylcellulose or hydroxymethylcellulose phthalate You may coat | cover with the film of a substance, and may coat | cover with two or more layers. Further, the capsule may be formed by a material such as gelatin or ethyl cellulose.
[0014]
When preparing a liquid composition for oral administration using the compound represented by the general formula [I] of the present invention as a main drug, a pharmaceutically acceptable emulsion, solubilizer, suspension, Dosage forms such as syrups or elixirs are possible. In this case, examples of the diluent used include purified water / ethanol, vegetable oil, and emulsifier. In addition to the diluent, these compositions may be mixed with an auxiliary agent such as a wetting agent, a suspending agent, a sweetening agent, a flavoring agent, a fragrance or a preservative.
[0015]
In the case of preparing an injection for parenteral administration using the compound represented by the general formula [I] of the present invention as a main drug, a sterile aqueous or non-aqueous solution, solubilizer, suspension or An emulsifier can be used. Examples of aqueous solutions, solubilizers, and suspensions include water for injection, distilled water for injection, physiological saline, cyclodextrin and derivatives thereof, and organic amines such as triethanolamine, diethanolamine, monoethanolamine, and triethylamine. Or there is an inorganic alkaline solution or the like.
[0016]
When the compound represented by the general formula [I] of the present invention is used as a main drug to form a water-soluble solution, for example, a vegetable oil such as propylene glycol, polyethylene glycol or olive oil, or an alcohol such as ethanol is used. It may be used. As the solubilizer, for example, surfactants such as polyoxyethylene hydrogenated castor oil and sucrose fatty acid ester (mixed micelle formation), lecithin or hydrogenated lecithin (liposome formation) and the like can be used. Furthermore, an emulsion preparation comprising a water-insoluble solubilizer such as vegetable oil and lecithin, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene glycol, or the like can be used.
Other compositions for parenteral administration include one or two or more kinds of main agents, that is, external preparations and ointments containing a compound represented by the above general formula [I] and the like and formulated by a method known per se Such a coating agent, suppository or pessary may be used.
[0017]
Examples Formulation examples in which the compound represented by the above general formula [I] of the present invention is used as an active ingredient of an anticancer agent will be specifically described below. However, the present invention is not limited by the following description.
[0018]
Example 1
In the injection preparation of the anticancer agent of this example, 1- [1-4- (diphenylmethyl) piperazinyl] -3- [1- [4- (4-chlorophenyl) -4-hydroxy] piperidinyl is used as the main drug. ] -2-propanol (hereinafter referred to as Compound 1) is used.
A synthesis example of the compound 1 will be described below. In the following description, the nuclear magnetic resonance spectrum (NMR) is measured using tetramethylsilane as an internal standard and is expressed in ppm. The part indicates a capacity part.
[0019]
(1) 1- dissolved (diphenylmethyl) -4- (1- (2,3-epoxy) propyl) piperazine emissions of 1- (diphenylmethyl) piperazine (10.0 g) in acetonitrile (50 ml), sodium carbonate ( 6.5 g) and epibromohydrin (6.8 g) were added, and the mixture was heated to reflux for 2.5 hours. After the salt was filtered off, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Wakogel C-200, 200 g) and eluted with a mixed solvent of 99 parts of chloroform and 1 part of methanol to give 1- (diphenylmethyl) -4- (1- (2,3 -Epoxy) propyl) piperazine (5.9 g) was obtained.
Nuclear magnetic resonance spectrum
¹ H-NMR (CDC13, 500 MHz) δ: 2.30 to 2.80 (12H, m), 3.06 to 3.10 (1H, m) 4.23 (1H, s), 7.16 (2H, t, J = 7.3 Hz), 7.25 (4H .T, J = 7.3 Hz), 7.40 (4H, d, J = 7.3 Hz).
[0020]
(2) Synthesis of 1- [1-4- (diphenylmethyl) piperazinyl] -3- [1- [4- (4-chlorophenyl) -4-hydroxy] piperidinyl] -2-propanol 1- (Diphenylmethyl)- 1. 4- (1- (2,3-epoxy) propyl) piperazine (3.0 g) and 4- (4-chlorophenyl) -4-hydroxypiperidine (2.5 g) are dissolved in o-dichlorobenzene (20 ml). The mixture was heated to reflux for 5 hours. After cooling, the mixture was purified by silica gel column chromatography (Wakogel C-200, 100 g), and 1- [1-4- (diphenylmethyl) piperazinyl] -3- [1- [4- (4-chlorophenyl) -4- 4.6 g of (hydroxy) piperidinyl] -2-propanol (Compound 1) were obtained.
Infrared absorption spectrum: IR ν max (cm −1) KBr: 3300, 2950, 2650, 1620, 1450, 1100, 910, 830, 750, 710 (as hydrochloride)
Nuclear magnetic resonance spectrum
¹ H-NMR (CDC13, 500 MHz) δ: 1.50 to 1.90 (4H, m), 2.01 to 2.21 (2H, m) 2.30 to 2.55 (10H, m), 2.80 to 2.90
(2H, m), 3.87 to 3.93 (1H, m), 4.22 (1H, s), 7.16 (2H, t, J = 7.3 Hz), 7.26 (4H.t, J = 7.3 Hz), 7.30 (2H, d, J = 8.5 Hz), 7.40 (4H, d, J = 7.3 Hz), 7.42 (2H, d, J = 8.5 Hz).
FD mass spectrum FD-MS (m / z): 519, 521 (M +).
[0021]
(1) Compound 1 Injection Formulation Compound 1 2-40 mg
D-sorbitol 1000mg
Citric acid 10mg
Sodium hydroxide Appropriate amount Water for injection Add water for injection to make 20.0 ml.
D-sorbitol and citric acid were dissolved in a sufficient amount of water for injection. Compound 1 was dissolved in the resulting solution, and the pH was adjusted to 3.2 to 3.3 with sodium hydroxide. The remaining water for injection was then added with stirring. The solution was filtered, filled into a 20.0 ml ampoule and sealed. The contents of this ampule were sterilized by autoclaving.
[0022]
(2) Examination of in vitro antitumor effect in various human cancer cell lines by MTT assay.
Human non-small cell lung cancer cell line PC-14, human small cell lung cancer cell line SBC-3, human breast cancer cell line MCF-7, human ovarian cancer cell line SKOV3, human leukemia cell line HL60 and human colon cancer cell line WiDR , RPMI, respectively
Single cells were prepared using trypsin treatment or cell scraper in 1640 medium, and 100 cell suspensions per 15 μl were prepared. To this, Compound 1 as an anticancer agent was dissolved in dimethyl sulfoxide and added so that the concentration of Compound 1 was in the range of 0.5 to 1 μM, and 150 μl per well was added to a 96-well plate. The plate was maintained for 96 hours and cultured under the condition of 5% carbon dioxide concentration and saturated water vapor at a temperature of 37 ° C. After culturing, MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (3- (4,5,5) dissolved in D-PBS (-) at a concentration of 5 mg / ml. -dimethy1thiazo1-2-y1) -2,5-diphenyltetrazoliumbromide)] reagent was added, and the mixture was further cultured at 37 ° C. for 4 hours. After completion of the culture, the whole plate was centrifuged and the supernatant was discarded. 200 μl of dimethyl sulfoxide is added, and yellow MTT dissolves purple formazan produced by the action of dehydrogenase in mitochondria in cancer cells, and the amount of produced formazan is determined by the absorbance at a wavelength of 562 to 630 nm. Was measured with a multiplate reader. A tumor volume growth curve was drawn with the average growth rate of the negative control being 0% and the average growth rate of the positive control being 100%, and the human cancer cell growth inhibitory concentration of Compound 1 was calculated. This is shown in Table 1.
[0023]
Table 1

The values in Table 1 are the mean ± error range (μM) of 50 percent cancer cell growth inhibitory concentration (IC ₅₀ ) value of MTT assay in 3 independent cancer cell culture experiments using Compound 1. It shows with.
[0024]
As shown in Table 1, Compound 1 exhibited a 50% growth inhibitory effect (IC ₅₀ ) for various cancers at a concentration of 0.5 to 1.1 μM. The IC ₅₀ value for Compound 1 is much lower than the IC ₅₀ value of 2 to 3 μM according to the MTT assay in cisplatin, which is said to be most effective for solid cancer. From the IC ₅₀ value, it can be seen that the anticancer effect of Compound 1 is large, and it is effective as an anticancer agent. In addition, solid tumor cells such as lung cancer, breast cancer, colorectal cancer, ovarian cancer and the like have a broad antitumor spectrum to show the same antitumor effect as leukemia cells.
[0025]
Example 2
Control, Compound 1 concentrations 1 × 10 - husband sequence of ^{6 M} series and compound 1 concentrations 3 × 10 ^-6 M s, was prepared nine dish (total of 27 sheets). To each of the 27 petri dishes, 4.4 × 10 ⁵ fibroblasts NIH3T3 were added per petri dish, and D-MEM culture medium supplemented with 10% FBS (fetal calf serum) was added to each of the 27 dishes. As a control, water corresponding to the amount of Compound 1 added was added and cultured. Every 24 hours, the number of cells in each series was measured and averaged to obtain the average number of cells per dish. The result is shown in FIG. As shown in the results, Compound 1 is effective against proliferative lesions such as interstitial pneumonia and keloid caused by fibroblast proliferation as well as anticancer activity.
[0026]
Example 3
Into female nude mice BALB / c nu / nu (6 weeks old), the cells 2 × 10 ⁷ cells of human non-small cell lung carcinoma lines PC-14 and physiological saline suspension, engraftment be implanted subcutaneously in the back The progress of was observed. Check the status of the tumor to 7 days after transplantation elect a test that can be individuals, and line Tsu randomizing of each individual so as not to cause deviation, respectively six addressed to the control series, the first test series and Divided into second test series. The tumor volume of each transplanted tumor is measured by measuring the tumor minor axis and the tumor major axis, and the product of the square of the tumor minor axis and the tumor major axis, ie: (tumor minor axis) ² × (tumor minor) The major axis was obtained. Administration of Compound 1 was started on day 7 after tumor implantation.
Compound 1 is dissolved in 1.2 mg of 0.05 ml of dimethyl sulfoxide, and 0.95 ml of 5% sorbitol-0.2% citric acid monohydrate solution (pH 3.3) is added to make a uniform solution. Compound 1 injection solution.
Compound 1 was administered to the 6 subjects in the series 1 subject group once a day on the 7th, 8th, 9th, 10th and 11th days after tumor transplantation. 3 mg of compound 1 per kg of body weight was injected from the tail of nude mice, and 6 animals in the group No. 2 subject group were on the 7th, 8th, 9th, and 10th days after tumor transplantation. On the eyes and on the 11th day, 5 mg of Compound 1 per kg of body weight was infused from the tail of nude mice once a day. The group of subjects with series number 1 as a control was a series in which compound 1 was not administered, and administration and treatment of compound 1 were not performed at all. For all groups of subjects, observation of the general condition of each mouse as well as once each day from the first day to the eighth day of administration of compound 1 and once every day before administration of compound 1 Measurements were taken of mouse body weight and tumor volume. In the observation and weight measurement result of the general condition of the mice was almost Tsu or a difference was observed between the sample group of sequence numbers 1 to 3, for the tumor volume of mice of the sequence numbers 1 to 3 Differences were seen between subject groups. The measurement results for the tumor volume are shown in the following Table 2, and the average value of the tumor volume of the subject group for each series number is shown in Table 3. In Tables 2 and 3, the size of the tumor volume after the second day is shown with the size of the tumor volume on the first day being 100.
[0027]
Table 2

[0028]
Table 3

1- [1-4- (diphenylmethyl) piperazinyl] -3- [1- [4- (4-chlorophenyl) -4-hydroxy] piperidinyl] -2-propanol From the results of Examples 1 to 3, compound 1 It can be seen that the anticancer effect by is great.
[0029]
In Examples 1 to 3, 1- [1-4- (diphenylmethyl) piperazinyl] -3- [1- [4- (4-chlorophenyl) -4-hydroxy] piperidinyl] -2-propanol of Compound 1 is 1- [2- (1,2,3,4-tetrahydro) isoquinolinyl] -3- [1- (4-diphenylmethyl) piperazinyl] -2-propanol as the main drug The same result was obtained.
[0030]
A synthesis example of 1- [2- (1,2,3,4-tetrahydro) isoquinolinyl] -3- [1- (4-diphenylmethyl) piperazinyl] -2-propanol will be described below. In the following description, the nuclear magnetic resonance spectrum (NMR) is measured using tetramethylsilane as an internal standard and is expressed in ppm. The part indicates a capacity part.
[0031]
(1) Synthesis of 2- [1- (2,3-epoxy) propyl] -1,2,3,4-tetrahydroisoquinoline 1,2,3,4-tetrahydroisoquinoline (25.0 g) in acetonitrile (100 ml) To this was added sodium carbonate (40.0 g) and epibromohydrin (31.0 g), and the mixture was heated to reflux for 4 hours. The salt was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Wakogel C-200, 500 g) and eluted with a mixed solvent of 99 parts of chloroform and 1 part of methanol to give 2- [1- (2,3-epoxy) propyl] -1 , 2,3,4-Tetrahydroisoquinoline (15.6
g) was obtained.
Nuclear magnetic resonance spectrum
¹ H-NMR (CDC13, 100 MHz) δ: 2.36 to 2.60 (2H, m), 2.73 to 3.03 (6H, m), 3.09 to 3.29 (1H, m), 3.65 (1H, d, J = 14.9 Hz) , 3.83 (1H, d,
[0032]
Example 4
1 - [2- (1,2,3,4-tetrahydro) isoquinolinyl] -3- [1- (4-diphenylmethyl) piperazinyl] -2-propanol synthesized 2- [1- (2,3-epoxy) Propyl) -1,2,3,4-tetrahydrisoquinoline (3.0 g) and 1- (diphenylmethyl) piperazine (4.4 g) are dissolved in o-dichlorobenzene (20 ml) and heated for 2.5 hours. Refluxed. The reaction mixture was allowed to cool and then purified by silica gel column chromatography (Wakogel C-200, 150 g) to give 1- [2- (1,2,3,4-tetrahydro) isoquinolinyl] -3- [1- (4-diphenylmethyl). was obtained piperazinyl] -2-propanol (6 .0g).
Infrared absorption spectrum: IR νmax (cm −1) KBr: 3400, 3000, 2550, 1620, 1450, 1080, 920, 760, 710 (as hydrochloride)
Nuclear magnetic resonance spectrum
¹ H-NMR (CDC 13, 100 MHz) δ: 2.30 to 2.60 ( ^{1, 2} H, m), 2.75 to 2.95 (4 H, m) 3.62 to 3.80 (2 H, m), 3.92 to 4.03 (1 H, m), 4.21 ( 1H, s), 7.00-7,51 (14H, m).
FD mass spectrum FD-MS (m / z): 441 (M +).
[0033]
【The invention's effect】
The compound represented by the general formula [I] according to the present invention is less toxic than a conventional anticancer agent such as cisplatin, and has various properties compared to a conventional anticancer agent. Compared to conventional anticancer drugs, cancer treatment for cancer (solid cancer) and other malignant tumors, etc. It makes a great contribution to the above. Further, the present invention has an action of suppressing the proliferation of fibroblasts such as pulmonary fibrosis, and has an excellent fibrosis-inhibiting action as compared with conventional therapeutic agents such as pulmonary fibrosis and keloid, It greatly contributes to the treatment of pulmonary fibrosis and keloid.
[Brief description of the drawings]
1 is a graph showing the cell proliferation inhibitory effect against fibroblasts shown to reduction compound 1 in Example 2.

Claims (3)

Formula [I]

[Wherein R is

Or

The anticancer agent characterized by containing the compound shown by these, or its salt .   The compound is 1- [1-4- (diphenylmethyl) piperazinyl] -3- [1- [4- (4-chlorophenyl) -4-hydroxy] piperidinyl] -2-propanol 1. The anticancer agent according to 1.   2. The compound according to claim 1, wherein the compound is 1- [2- (1,2,3,4-tetrahydro) isoquinolinyl] -3- [1- (4-diphenylmethyl) piperazinyl] -2-propanol. The anticancer agent described.

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