JP2005500365A - C-2 'methylated derivative of paclitaxel for use as an antitumor agent - Google Patents

Abstract

A taxane derivative of formula (I), wherein R is trifluoromethyl, phenyl, 2-furyl, 2-thienyl; R ¹ is t-butoxycarbonyl or benzoyl; R ₂ is hydroxy R ₃ together with hydrogen or R ₂ forms the residue of a cyclic carbonate of formula II, provided that when R ₃ is hydrogen, R is not phenyl. The compounds of formula (I) have antitumor activity.

Description

【０００４】
〔式中、
Ｒは、トリフルオロメチル、フェニル、２−フリル、２−チエニルであり；
Ｒ¹は、ｔ−ブトキシカルボニル又はベンゾイルであり；
Ｒ₂は、ヒドロキシであり；
Ｒ₃は、水素又は、Ｒ₂と一緒に、式：
【０００５】
【化４】

【０００６】
で示される環状カルボナートの残基を形成するが、但し、Ｒ₃が、水素である場合、Ｒは、フェニルではない〕
で示される化合物に関する。
【０００７】
式（Ｉ）の化合物は、抗腫瘍活性を有する公知の医薬品、パクリタキセル又はドセタキセルの誘導体である。
【０００８】
Ｒ₃が、水素であり、Ｒが、フェニルである式（Ｉ）の化合物は、Greene らにより、J. Chem. Soc. Perkin Trans.l, 1995, pp 1811-1815に述べられている。Ｃ−２′−メチル化（２Ｒ，３Ｓ）側鎖を有するパクリタキセル誘導体は、Kantらにより、Tetrahedron, vol. 37, n. 36, pp 6495-6498, 1996に述べられている。
【０００９】
Kantは、微小管に対するより優れた結合親和性とKBVIに対するより優れた細胞毒性のために、この構造修飾が、特定の試験（微小管脱重合の阻害）において、親化合物であるパクリタキセルと比較して、有意な応答の増強をもたらすことを示した。加えて、１４β−ヒドロキシバッカチン １，１４−カルボナートから合成されたこれらのタキサンの一部は、水溶性が改善されていた。最後に、チトクロームＰ−４５０クラスの酵素に関連する代謝経路を阻害するために、これらのタキサンの一つは、Ｃ−３位にトリフルオロメチル置換基を含む。これらの新規化合物の改善された薬理学的な特徴は、様々な腫瘍のタイプに対する活性スペクトルの修飾に良好に相関するだろう。
【００１０】
本発明の好ましい化合物は、以下のものである：
(２′Ｒ，３′Ｒ)−１３−［Ｎ−ベンゾイル−３−（２−フリル）−２−メチル−イソセリノイル］−バッカチンIII；
（２′Ｒ，３′Ｒ）−１３−［Ｎ−ｔ−ブトキシカルボニル−３−（２−フリル）−２−メチル−イソセリノイル］−バッカチンIII；
（２′Ｒ，３′Ｓ）−１３−［Ｎ−ｔ−ブトキシカルボニル−３−フェニル−２−メチル−イソセリノイル］−１４β−ヒドロキシバッカチンIII １，１４−カルボナート；
（２′Ｒ，３′Ｒ）−１３−［Ｎ−ｔ−ブトキシカルボニル−３−トリフルオロメチル−２−メチル−イソセリノイル］−１４β−ヒドロキシバッカチンIII １，１４−カルボナート；
（２′Ｒ，３′Ｒ）−１３−［Ｎ−ｔ−ブトキシカルボニル−３−（２−フリル）−２−メチル−イソセリノイル］−１４β−ヒドロキシバッカチンIII １，１４−カルボナート；
（２′Ｒ，３′Ｒ）−１３−［Ｎ−ｔ−ブトキシカルボニル−３−（２−チエニル）−２−メチル−イソセリノイル］−１４−ヒドロキシバッカチンIII １，１４−カルボナート。
【００１１】
式（Ｉ）の化合物は、式（II）：
【００１２】
【化５】

【００１３】
（式中、Ｒ₂及びＲ₃は、上記で定義したとおりであり、そしてＲ₄は、保護基、好ましくはトリエチルシリルである）
で示される化合物と、式（III）：
【００１４】
【化６】

【００１５】
（式中、Ｒ及びＲ₁は、上記で定義したとおりである）
で示される化合物との反応によって製造される。
【００１６】
式（III）の化合物を、アルコール溶媒中の加水分解によって、対応するアルキルエステルから、特に、Tetrahedron Asymm. 2001, 12, 1015-1027及びJ. Chem. Soc. Perkin Trans.l 1995, 1811-1816により公知のメチルエステルから製造することができる。得られた酸を、単離することなく、適切な溶媒中で、適切な縮合剤、例えばチオノ炭酸ジ−２−ピリジル及びジメチルアミノピリジンの存在下に、式（II）のバッカチンIII誘導体と直接に縮合することができる。７−位のヒドロキシ保護基を、所望の式（Ｉ）の化合物を得るために除去する。
【００１７】
あるいは、式（II）の化合物を、式（IV）：
【００１８】
【化７】

【００１９】
（式中、Ｒは、上記で定義したとおりである）
で示される化合物と反応させてもよい。
【００２０】
Ｒが、フェニルである式（IV）の化合物は、J. Org. Chem. 1991, 56, 1681-1684；Tetrahedron, 1992, 48, 6585-7012；EP 400971；US 5,175,315に記載されている。
【００２１】
Ｒが、トリフルオロメチルである式（IV）の化合物は、以下のスキームに従い、製造されうる。
【００２２】
【化８】

【００２３】
Ｒが、２−フリルである式（IV）の化合物を、J. Org. Chem. 1999, 64, 4643-4651 により公知の、（３Ｒ，４Ｒ）−４−（フラン−２−イル）−３−ヒドロキシ−３−メチル−アゼチジン−２−オンから出発して、以下のスキームに従って、製造することができる。
【００２４】
【化９】

【００２５】
式（II）の化合物は、J. Med. Chem. 1997, 40, 267-278により公知である。
【００２６】
化合物（II）と化合物（IV）との反応は、不活性雰囲気下に、非プロトン性溶媒中で、実施される。
【００２７】
典型的には、この反応は、ヘキサメチルジシラザンナトリウムの存在下に、約−４０℃〜５℃の温度で実施される。次いで、７位のヒドロキシ保護基を除去し、式（Ｉ）の化合物を与える。
【００２８】
本発明の化合物は、乳房、肺、卵巣、結腸、前立腺、腎臓、すい臓の腫瘍細胞に対して、並びにアドリアマイシン、ビンブラスチン及び白金誘導体のような公知の抗腫瘍薬に耐性のある細胞に対して強力な抗腫瘍活性を示した。
【００２９】
したがって、本発明は、薬理学的に許容されうる担体及び賦形剤と共に、有効量の本発明の化合物を含む医薬製剤に関する。さらに特には、化合物を、錠剤、粉剤、顆粒剤、カプセル剤、注射溶液、坐剤、乳剤、分散剤などの形態に調製してもよい。静脈内投与のために、Chremophor L及びエタノール、ポリソルベート及びエタノール、又は天然若しくは合成のホスファチジルコリンにより調製したリポソーム製剤の混合物、あるいはコレステロール存在下の天然のリン脂質の混合物が、好ましく用いられ；経口投与のために、ソフトゼラチンカプセルが、場合によりリン脂質の存在下に、生成物が、ポリソルベート、ＰＥＧ又はそれらの混合物で可溶化されるように、好ましく調製される。化合物（Ｉ）は、５０〜５００mg／m²の範囲の濃度で、ヒトに投与されうる。
【００３０】
以下の実施例は、より詳細に本発明を説明する。
【００３１】
用いた略号は、以下の通りである：
ＴＥＳ＝トリエチルシリル；ＤＭＦ＝ジメチルホルムアミド；ＤＭＡＰ＝（Ｎ，Ｎ−ジメチルアミノ）ピリジン；ＮａＨＭＤＳ＝ナトリウム ヘキサメチルジシラジド；ＬｉＨＭＤＳ＝リチウム ヘキサメチルジシラジド；ＴＨＦ＝テトラヒドロフラン；ＨＭＰＡ＝ヘキサメチルリン酸トリアミド。
【００３２】
実施例１
メタノール中の（４Ｓ，５Ｒ）−３−ベンゾイル−２−（２，４−ジメトキシフェニル）−４−（フラン−２−イル）−５−メチルオキサゾリジン−５−カルボン酸メチルエステル（０．１２８g、０．２８５mmol）を、Ｋ₂ＣＯ₃（２eq）と無水媒体中で、２５℃で２４時間撹拌した。反応混合物を減圧下に濃縮し、飽和ＮＨ₄Ｃｌ溶液で希釈し、酢酸エチルで抽出した。水相を５％ＮａＨＳＯ₄でｐＨ４に酸性化し、酢酸エチルで抽出した。有機相を乾燥させ、そして溶媒を蒸発除去した。残渣（１eq）を、アルゴン雰囲気下、２０℃で、チオノ炭酸ジ−２−ピリジル（１eq）及びジメチルアミノピリジン（０．５eq）の存在下に、７−ＴＥＳ−バッカチンIII（０．０４g、０．０５７mmoｌ）のトルエン溶液に加えた。反応混合物を、撹拌しながら８０時間、６０℃で加熱した。酢酸エチルの添加後、反応混合物をブラインで抽出した。有機相をブラインで洗浄し、次いで乾燥させ、蒸発させた。クロマトグラフィー（ＳｉＯ₂、酢酸エチル／ｎ−へキサン、１：１）により、（２′Ｒ，３′Ｒ）−１３−［Ｎ−ベンゾイル−Ｎ，Ｏ−（２，４−ジメトキシベンジリデン）−３−（２−フラニル）−メチルイソセリノイル］−７−ＴＥＳ−バッカチンIII（０．０２３g、２．２８mmol、４０％）を得た。
【００３３】
この化合物は、以下の特性を有する：
¹H NMR (CDC1₃) : δ＝0.61 (m, 6H, 3 CH₂), 0.95 (t, 9H, 3 Me), 1.22 (s, 3H, Me), 1.27 (s, 3H, Me), 1.68 (s, 3H, Me), 1.89 (m, 1H of C6-H), 2.10-2.25 (m, 2H of C14-H), 2.21 (s, 3H, Me), 2.24 (s, 3H, Me), 2.33 (s, 3H, Me), 2.50 (m, 1H of C6-H), 2.65 (s, 3H, Me, OAc of C-4), 3.90-4.0 (m, 8H, 6H of 2 Me,1H of C3-H, and 1H of OH), 4.12 (d, 1H of C20-H, J = 8.0 Hz), 4.32 (d, 1H, C20-H), 4.58 (m, 1H of C7-H,J_l = 5.8 Hz, J₂ = 10.2 Hz), 4.95 (dd, 1H of C5-H, J₁ = 1.5 Hz, J₂ = 9.6 Hz), 5.55 (b, 1H of C3'-H), 5.70 (d, 1H of C2-H,J=5.7 Hz), 6.30-6.40 (m, 2H), 6.40-6. 50 (m, 3H), 6.54 (s,1H), 6.82 (s, 1H), 7.20-7.40 (m, 5H, arom), 7.40-7.50 (m, 3H, arom), 7.54-7.60 (m, 1H, arom), 8.04-8.07 (m, 2H, arom)。
【００３４】
この化合物（０．０２３g、２．２８mmol）を、ＣＨ₂Ｃ１₂（３．０ml）に溶解し、２５℃で、塩化アセチルのメタノール（ＭｅＯＨ１０ml中に、塩化アセチル７０μl）溶液（１００μl）により処理した。７時間後、反応混合物を水で抽出した。有機相を乾燥させ、そして蒸発させた。残渣をクロマトグラフィー（ＳｉＯ₂、ｎ−へキサン／酢酸エチル、１：１）により分離し、０．０１９g（０．０２２mmol、９８％）の（２′Ｒ，３′Ｒ）−３′−デフェニル−３′−（２−フリル）−２′−メチル−パクリタキセルを得た。
【００３５】
この化合物は、以下の特性を有する：
¹H NMR (CDC1₃) : δ＝1.13 (s, 3H, Me), 1.23 (s, 3H, Me), 1.55 (s, 3H, Me), 1.70 (s, 3H, Me), 1.81 (s, 3H, Me), 1.89(m, 1H, H-6β), 2.07(m, 1H of C14-H), 2.24 (s, 3H, Me), 2.40 (m, 1H of C14-H), 2.49 (d, 1H, OH of C7-H), 2.55 (m, 1H of C6-H), 2.65 (s, 3H, Me), 3.75 (s, 1H, OH), 3.82 (d, 1H of C3-H, J = 7.2 Hz), 4.21 (d, 1H of C20-H J = 8.4 Hz), 4.31 (d, 1H of C20-H), 4.40 (m, 1H of C7-H), 4.95 (dd, 1H of C5-H, J₁ = 1.5 Hz, J₂ = 9.6 Hz), 5.66 (d, 1H of C2-H), 5.79 (d, 1H, of C3'-H, J = 9.5 Hz), 6.26 (s, 1H of C10-H), 6.30(m, 1H of C13-H), 6.41(m, 2H, 2-フリル), 7.12 (d, 1H, NH), 7.33 (m, 2H, arom), 7.44 (m, 1H, 2-フリル), 7.46 (m, 1H, arom), 7.52 (m, 2H, arom), 7.60 (m, 1H, arom), 7.67 (m, 2H, arom), 8.19 (m, 2H, arom)。
【００３６】
実施例２
実施例１に記載された手順に従って、（４Ｓ，５Ｒ）−Ｎ−Ｂｏｃ−Ｎ，Ｏ−（２，４−ジメトキシベンジリデン） −３−フェニル−２−メチルイソセン メチルエステル（J. Chem. Soc. Perkin Trans. 1995, 1811-1816）（０．１５４g、０．３３６mmol）と、７−ＴＥＳ−バッカチンIII １，１４−カルボナート¹ （０．０５g、０．０６７mmol）とのカップリングにより、０．０２１g（０．０２３mmol、３４％）の（２′Ｒ，３′Ｓ）−１３−（Ｎ−Ｂｏｃ−２−メチル−３−フェニルイソセリノイル）−１４β−ヒドロキシバッカチンIII １，１４−カルボナートを得た。
【００３７】
この化合物は、以下の特性を有する：
¹H NMR (CDC1₃) : δ＝1.28 (s, 9H, 3 Me of t-Boc), 1.30 (s, 3H, Me of C-15), 1.38 (s, 3H, Me of C-15), 1.41 (s, 3H, Me of C2'), 1.73 (s, 3H, Me of C-8), 1.84 (s, 3H, Me of C-12), 1.92 (m, 1H of C6-H), 2,26 (s, 3H, Me, OAc of C-10), 2.36 (b, 1H, OH), 2.56 (m, 1H of C6-H), 2.69 (s, 3H, Me, OAc of C-4), 3.52-3.55 (b, 1H, OH), 3.73 (d, 1H of C3-H, J = 7.3 Hz), 4.25 (d, 1H of C20-H, J = 8.5 Hz), 4.31 (d, 1H, of C20-H), 4.38 (m, 1H of C7-H), 4.85 (d, 1H of C14-H, J = 7.1 Hz), 4.95 (dd, 1H of C5-H, J_l = 2.4 Hz, J₂ = 9.6 Hz), 5.09 (d, 1H of C3'-H), 5.57 (d, 1H, NH, J = 10.0 Hz), 6.13 (d, 1H of C2-H), 6,27 (s, 1H of C10-H), 6.36 (m, 1H of C13-H), 7.30-7.40 (m, 5H, arom), 7.48 (m, 2H, arom), 7.59 (m, 1H, arom), 8.04 (m, 2H, arom)。
【００３８】
実施例３
ａ）（３Ｒ，４Ｓ）−３−トリエチルシリルオキシ−４−（フラン−２−イル）−３−メチル−アゼチジノン
塩化トリエチルシリル（０．３１６g、２．１mmol）及びイミダゾール（０．１００g、１．５mmol）を、アルゴン雰囲気下、２５℃で、ＤＭＦ（６．０ml）中の（３Ｒ，４Ｒ）−４−（フラン−２−イル）−３−ヒドロキシ−３−メチル−アゼチジン−２−オン（０．１６７g、１．０mmol）の撹拌溶液に加えた。反応溶液を、４時間後に、飽和ＮＨ₄Ｃｌ水溶液でクエンチし、酢酸エチルで抽出した。有機相を乾燥させ、減圧下に濃縮した。残渣のクロマトグラフィー（ＳｉＯ₂、酢酸エチル／ｎ−ペンタン、１：２）により、０．１９０g（０．６７mmol、６７％）の（３Ｒ，４Ｓ）−４−（フラン−２−イル）−３−トリエチルシリルオキシ−３−メチル−アゼチジン−２−オンを得た。
【００３９】
[α]_D ²⁰ ＝ + 42.8 (c 1.04, CHCl₃)；
IR (CDCl₃, cm^-1) : 3600-3000, 3413, 2957, 1768, 1458, 1377, 1012；
¹H NMR (CDCl₃) : δ＝0.65 (m, 6H, 3 CH₂), 0.95 (t, 9H, 3 Me), 1.19 (s, 3H, Me), 4.54 (s, 1H), 6.27 (d, 1H, 2-フリル), 6.28 (m, 1H, 2-フリル), 6.72 (b, 1H, NH), 7.40 (m, 1H, 2-フリル)；
¹³H NMR (CDCl3) : δ＝5.71 (CH₂), 6.73 (Me), 19.3 (Me), 61.0 (CH), 89.1 (C), 108.1 (CH), 110.5 (CH), 142.7 (CH), 150.9 (C), 171.5 (C)。
【００４０】
ｂ）（３Ｒ，４Ｓ）−１−tert−ブトキシカルボニル−３−トリエチルシリルオキシ−４−（フラン−２−イル）−３−メチル−アゼチジノン
前工程からの誘導体（０．０６０g、０．２１mmol）、ＤＭＡＰ（０．０１０g）、及びトリエチルアミン（８８μl、０．６３mmol）のＣＨ₂Ｃ１₂（３．０ml）溶液に、０℃で、二炭酸ジ−tert−ブチル（１２１ml、０．５２mmol）のＣＨ₂Ｃ１₂（１．０ｍｌ）溶液を加えた。反応混合物を、２５℃で１時間撹拌し、次いで飽和ＮＨ₄Ｃｌでクエンチした。混合物を、酢酸エチルで抽出し、有機層をブラインで洗浄し、乾燥させ、そして減圧下に濃縮した。クロマトグラフィー（ＳｉＯ₂、ｎ−ペンタン／酢酸エチル、８：２）により、０．０７７g（０．０２０mmol、９７％）の（３Ｒ，４Ｓ）−１−tert−ブトキシカルボニル−３−トリエチルシリルオキシ−４−（フラン−２−イル）−３−メチル−アゼチジノンを得た。
【００４１】
[α]_D ²⁰ ＝ + 26.6 (c 0.98，CHCl₃)；
IR (CDCl₃, cm^-1): 2958, 1813, 1726, 1327, 1152;
¹H NMR (CDC1₃) : δ＝0.54 (m, 6H,3 CH₂), 0.80 (t, 9H, 3 Me), 1.41 (s, 9H, 3 Me), 1.61 (s, 3H, Me), 4.73 (s, 1H), 6.27 (d, 1H, 2-フリル), 6.34 (m, 1H, 2-フリル), 7.37 (m, 1H, 2-フリル)
¹³H NMR (CDC1₃): δ＝5.6 (CH₂), 6.5 (Me), 23.0 (Me), 27.8 (3 Me), 62.7 (CH), 83.3 (C), 85.4 (C), 108.9 (CH), 110.3 (CH), 142.4 (CH), 147.9 (C), 148.7 (C), 167.7 (C)。
【００４２】
ｃ）（２′Ｒ，３′Ｒ）−１３−［Ｎ−Ｂｏｃ−２−メチル−３−（２−フラニル）−イソセリノイル］−バッカチンIII
工程ｂ）からのβ−ラクタム（０．０８２g、０．２１４mmol）及び７−ＴＥＳ−バッカチンIII（０．０６０ｇ、０．０８６mmol）を、アルゴン雰囲気下、ＴＨＦに溶解した。溶液を、−４５℃に冷却した。ＮａＨＭＤＳ（１．０Ｍ ｎ−ヘキサン溶液、２．５eq）を撹拌しながら、一滴ずつ加えた。温度を、４時間で−２０℃まで上げた。反応を、飽和ＮＨ₄Ｃｌでクエンチし、混合物を、酢酸エチルで抽出し、乾燥させた。溶媒の蒸発後、未精製物を、クロマトグラフィーにより精製した。反応生成物を、０℃で、１：１ ＭｅＣＮ／ピリジン溶媒に溶解した。ＨＦ／ピリジン（７０／３０）の溶液を、一滴ずつ（０．１ml／１０mgの試薬）を滴下した。反応混合物を、０℃で、１時間撹拌し、次いで２５℃で、６時間撹拌し、そして酢酸エチルで抽出した。抽出物を、飽和ＣｕＳＯ₄及び水で３回洗浄し、無水ＭｇＳＯ₄で乾燥させ、表題生成物を０．０３５g（０．０４１mmol、４８％）得た。
【００４３】
¹H NMR (CDC1₃) : δ＝1.15 (s, 3H, Me a C-15), 1.23 (s, 9H, 3 Me of t-Boc), 1.30 (s, 3H, Me a C-15), 1.42 (s, 3H, Me a C2'), 1.69 (s, 3H, Me a C-8), 1.84 (s, 3H, Me a C-12), 1.89 (m, 1H, H-6β), 2.15 (m, 1H, H-14), 2,25 (s, 3H, Me, OAc of C-10), 2.37 (m, 1H, H-14), 2.55 (m, 1H of C6-H), 2.61 (s, 3H, Me, OAc of C-4), 3.58-3.65 (b, 1H, OH), 3.82 (d, 1H of C3-H, J = 7.0 Hz), 4.18 (d, 1H of C20-H, J = 8.4 Hz), 4.31 (d, 1H of C20-H), 4.42 (dd, 1H of C7-H, J_l = 6.4 Hz, J₂ = 10.8 Hz), 4.95 (dd, 1H of C5-H, J₁ = 2.4 Hz, J₂ = 9.6 Hz), 5.23 (d, 1H, J = 9.6 Hz, NH), 5.39 (d, 1H of C3'-H), 5.66 (d, 1H of C2-H), 6.28 (s, 1H, of C10-H), 6.34 (m, 1H, 2-フリル), 6.36 (m, 1H, H-13), 6.38 (m, 1H, 2-フリル), 7.42 (m, 1H, 2-フリル), 7.49 (m, 2H, arom), 7.59 (m, 1H, arom), 8.14 (m, 2H, arom)。
【００４４】
実施例４
（２′Ｒ，３′Ｒ）−１３−［Ｎ−Ｂｏｃ−２−メチル−３−（２−フラニル）イソセリノイル］−１４β−ヒドロキシバッカチンIII １，１４−カルボナート
７−ＴＥＳ−１４β−ヒドロキシバッカチンIII １，１４−カルボナート（０．０８０g、０．１２５mmol）を、実施例３ｃに記載した条件で、（３Ｒ，４Ｓ）−ｌ−tert−ブトキシカルボニル−３−トリエチルシリルオキシ−４−（フラン−２−イル） −３−メチル−アゼチジノン（０．１２g、０．３１２mmol）とカップリングさせ、（２′Ｒ，３′Ｓ）−１３−［Ｎ−Ｂｏｃ−２−メチル−３−（２−フラニル）イソセリノイル］−３′，７−ジＴＥＳ−１４β−ヒドロキシバッカチンIII １，１４−カルボナート（０．０８７g、０．０７７５mmol、６２％）を得た。
【００４５】
¹H NMR (CDC1₃) : δ＝0.5-0.6 (m, 12H, 6 CH₂), 0.85-0.95 (m, 18H, 6 Me), 1.25 (s, 3H, Me), 1.29 (s, 9H, 3 Me), 1.31 (s, 3H, Me), 1.56 (s, 3H, Me), 1.75 (s, 3H, Me), 1.92 (m, 1H of C6-H, J_6-6, = 14.3 Hz), 2.02 (s, 3H, Me), 2.20 (s, 3H, Me), 2.51 (m, 1H of C6-H), 2.71 (s, 3H, Me), 3.76 (d, 1H of C3-H, J= 8.1 Hz), 4.26 (q, 2H of C20, J= 8.5 Hz), 4.44 (m, 1H of C7-H, J_l = 10.8 Hz, J₂ = 7.0 Hz), 4.85 (d, 1H of C14-H, J = 7.0 Hz), 4.89 (m, 1H of C5-H, J₁ = 1.8 Hz, J₂ = 9.8 Hz), 5.24 (d, 1H, 1H of C3'-H, J = 10.0 Hz), 5.28 (d, 1H of NH), 6.10 (d, 1H of C2-H), 6.26 (d, 1H of 2-フリル), 6.38 (m, 1H of 2-フリル), 6.42 (s, 1H of C10-H), 6.47 (d, 1H of C13-H), 7.38 (m, 1H of 2-フリル), 7.42-7.50 (m, 2H, arom), 7.54-7.60 (m, 1H, arom), 8.02-8.08 (m, 2H, arom)。
【００４６】
このような誘導体を、ＨＦ／ピリジン溶液で処理することにより、表題化合物（０．０６３g、０．７０mmol、９１％）を得た。
【００４７】
¹H NMR (CDC1₂) : δ1.25 (s, 9H, 3 Me), 1.28 (s, 3H, Me), 1.43 (s, 3H, Me), 1.75 (s, 3H, Me), 1.86 (s, 3H, Me), 1.92 (m, 1H of C6'-H, J_6-6' = 14.3 Hz), 2.24 (s, 3H, Me), 2.55 (m, 1H of C6-H), 2.63 (s, 3H, Me), 3.73 (d, 1H of C3-H, J = 8.0 Hz), 4.26 (q, 2H of C20, J = 8.5 Hz), 4.38 (m, 1H of C7-H, J₁ = 10.8 Hz, J₂ = 7.0 Hz), 4.86 (d, 1H of C14-H, J = 7.0 Hz), 4.93 (m,1H of C5-H, J_l = 1.8 Hz, J₂ = 9.8Hz), 5.26 (d, 1H, 1H of C3'-H, J = 9.5 Hz), 5.44 (d, 1H of NH), 6.12 (d, 1H of C2-H), 6.28 (s, 1H of C10-H), 6.36 (d, 1H of 2-フリル), 6.40 (m, 1H of 2-フリル), 6.47 (d, 1H of C13-H), 7.42 (m, 1H of 2-フリル), 7.44-7.50 (m, 2H, arom), 7.58-7.62 (m, 1H, arom), 8.02-8. 08 (m, 2H, arom)。
【００４８】
実施例５
ａ）（３Ｒ，４Ｒ）−３−ヒドロキシ−１−（４−メトキシフェニル）−３−メチル−４−トリフルオロメチル−アゼチジン−２−オン
（J. Org. Chem., 1999, 64, 4643-4651に記載されている）（２Ｓ，５Ｓ）−２−tert−ブチル−２，５−ジメチル−［１，３］ジオキソラン−４−オン（０．３４４g、２．０３mmol）のＴＨＦ溶液に、ＬＩＨＭＤＳの１Ｍ ＴＨＦ溶液（２．４ml、２．４mmol）を−７８℃で加えた。３０分後、ＨＭＰＡ及びＮ−（４−メトキシフェニル）トリフルオロアセタールジミン（０．８１g，４.００mmol）の溶液を連続して加えた（ＴＨＦ：ＨＭＰＡ＝８５：１５）。４時間後、反応混合物を、−７８℃で、１Ｍ ＣＨ₃ＣＯＯＨ水溶液５mlにより処理し、１Ｎ ＨＣｌ、次いでＮＨ₄Ｃｌ、最後にブラインで抽出した。有機相を乾燥させ（Ｎａ₂ＳＯ₄）、溶媒を減圧下に蒸発除去した。残渣のクロマトグラフィー（ＳｉＯ₂、酢酸エチル／ｎ−ペンタン、１：２）により、表記のβ−ラクタム（３Ｒ，４Ｒ）を０．２５g（０．９１mmol、４５％）得た。
【００４９】
[α]_D ²⁰ ＝ + 28.4 (c 1.01，CHCl₃)；
¹H NMR (CDC1₃) : δ＝1.70 (d, 3H, Me, J = 1.2 Hz), 3.02-3.10 (b, 1H, OH), 3.78 (s, 3H, OMe), 4.33 (q, 1H, J_H-F = 5.7 Hz), 6.85-7.40 (m, 4H, arom);
¹³H NMR (CDC1₃): δ＝22.2 (Me), 55.5 (OMe), 63.9 (q, CH, J = 31 Hz), 82.8 (C), 114.4 (2 CH), 119.7 (2 CH), 123.7(CF₃, J = 279 Hz), 129.3 (C), 157.3 (C), 167.9 (C)。
【００５０】
ｂ）（３Ｒ，４Ｒ）−３−トリエチルシリルオキシ−１−（４−メトキシ−フェニル）−３−メチル−４−トリフルオロメチル−アゼチジン−２−オン
ａ）からの化合物（０．２５g、０．９１mmol）のＤＭＦ（３．０ml）溶液に、２５℃でＥｔ₃ＳｉＣｌ（０．３１g、２．０mmol）及びＮ−メチルイミダゾール（０．２８g、４mmol）を加えた。２時間の撹拌後、反応混合物を氷水に注加し、酢酸エチルで抽出し、乾燥させた。溶媒を蒸発除去し、残渣をクロマトグラフィーに付し、表題生成物を０．３２g（０．８２mmol、９０％）得た。
【００５１】
IR(CDCl₃, cm^-1) : 2957, 2878, 1778, 1514, 1298, 1251;
¹H NMR (CDC1₃) : δ＝0.75 (m, 6H, 3 CH₂), 0.98 (t, 9H, 3 Me), 1.65 (s, 3H, Me), 3.79 (s, 3H, OMe), 4.22 (q, 1H, J_H-F = 5.9 Hz), 6.85-7.40 (m, 4H, arom);
¹³H NMR (CDC1₃): δ＝5.7 (CH₂), 6.6 (Me), 23.4 (Me), 55.5 (OMe), 64.3 (q, CH, J = 32 Hz), 84.0 (C), 114.4 (2 CH), 119.5 (2 CH), 123.6(CF₃, J = 283 Hz), 129.8 (C), 157.1 (C), 167.1 (C)。
【００５２】
ｃ）（３Ｒ，４Ｒ）−３−トリエチルシリルオキシ−３−メチル−４−トリフルオロメチル−アゼチジン−２−オン
ｂ）からの化合物（０．３０g、０．７７mmol）のアセトニトリル（１３．０ml）溶液に、−５０℃、２時間で、水２０．０ml中の硝酸アンモニウムセリウム（IV）（１．５g、２．７４mmol）、さらに水３０mlを一滴ずつ加えた。混合物を、水３０mlで希釈し、酢酸エチルで抽出した。有機相を飽和ＮａＨＣＯ₃、飽和ＮａＨＳＯ₃、そして再度飽和ＮａＨＣＯ₃で洗浄した。有機層を乾燥し、減圧濃縮した。残渣をカラムクロマトグラフィー（ＳｉＯ₂、ＣＨ₂Ｃｌ₂／酢酸エチル、３：１）に付し、表題化合物（０．１６８g、０．５９mmol、７７％）を得た。
【００５３】
¹H NMR (CDC1₃) : δ＝0.69 (m, 6H, 3 CH₂), 0.94 (t, 9H, 3 Me), 1.60 (s, 3H, Me), 3.77 (q, 1H, J_H-F = 6.2 Hz), 6.20-6.45 (b, 1H, NH);
¹³H NMR (CDC1₃): δ＝5.8 (CH₂), 6.8 (Me), 23.7 (Me), 61.0 (q, CH, J = 32 Hz), 86.3 (C), 123.8(CF₃, J = 280 Hz), 170.8 (C)。
【００５４】
ｄ）（３Ｒ，４Ｒ）−１−（tert−ブトキシカルボニル）−３−トリエチルシリルオキシ−３−メチル−トリフルオロメチル−アゼチジン−２−オン
ｃ）からの化合物（０．１６８g、０．５９mmol）、ＤＭＡＰ（１０mg）及びトリエチルアミン（０．２５ml、１．７７mmol）のジクロロメタン（２．０ml）溶液に、０℃で、二炭酸ジ−tert−ブチル（０．３２g、１．４７mmol）のジクロロメタン（１．０ml）溶液を加えた。反応混合物を、２５℃で２時間撹拌し、次いで飽和ＮＨ₄Ｃｌでクエンチした。混合物を酢酸エチルで抽出し、有機層をブラインで洗浄し、乾燥させ、そして減圧下に濃縮した。クロマトグラフィー（ＳｉＯ₂、ｎ−ペンタン／酢酸エチル、４：１）により表題生成物０．２１０g（０．５６mmol、９５％）を得た。
【００５５】
¹H NMR (CDC1₃) : δ＝0.69 (m, 6H, 3 CH₂), 0.95 (t, 9H, 3 Me), 1.52 (s, 9H, 3 Me), 1.63 (s, 3H, Me), 4.10 (q, 1H, J_H-F = 6.2 Hz);
¹³H NMR (CDC1₃): δ＝5.6 (CH₂), 6.5 (Me), 23.5 (Me), 27.8 (3 Me), 63.2 (q, CH, J= 32 Hz), 84.7 (C), 122.8 (CF₃, J = 281 Hz), 147.2 (C), 167.4 (C)。
【００５６】
ｅ）（２′Ｒ，３′Ｒ）−１３−［Ｎ−Ｂｏｃ−２−メチル−３−トリフルオロメチルイソセリノイル］−１４β−ヒドロキシバッカチンIII １，１４−カルボナート
実施例４の手順に従って、７−ＴＥＳ−１４β−ヒドロキシバッカチンIII（０．０８０g、０．１１mmol）と、ｄ）からの化合物（０．１０３g、０．２７mmol）との反応により、表題化合物の７，３′−ジ−ＴＥＳ−誘導体（０．０７１g、０．０６３mmol、５７％）を得た。
【００５７】
¹H NMR (CDC1₃) : δ＝0.62-0.72 (m, 12H, 6 CH₂), 0.88-0.95 (m, 18H, 6 Me), 1.25 (s, 9H, 3 Me), 1.31 (s, 3H, Me), 1.52 (s, 6H, 2 Me), 1.63 (s, 3H, Me), 1.90 (m, 1H of C6'-H), 2.00 (s, 3H, Me), 2.19 (s, 3H, Me), 2.48 (m, 1H of C6-H), 2.58 (s, 3H, Me), 3.74 (d, 1H of C3-H, J = 7.5 Hz), 4.24 (q, 2H of C20, J = 8.8 Hz), 4.41 (m, 1H of C7-H, J_l = 10.6 Hz, J₂ = 6.4 Hz), 4.74 (m, 1H, C3'-H), 4.83 (d, 1H of C14-H, J = 6.9 Hz), 4.86 (m, 1H of C5-H, J_l = 1.90Hz, J₂ = 9.8 Hz), 5.10 (d, 1H of NH), 6.10 (d, 1H of C2-H), 6.40 (s, 1H of C10-H), 6.42 (m, 1H of C13-H), 7.44-7.50 (m, 2H, arom), 7.58-7.60 (m,1H, arom), 8.06-8.10 (m, 2H, arom)。
【００５８】
この誘導体を、ＨＦ／ピリジン溶液で処理し、表題化合物（０．０４８g、０．５４mmol、８５％）を得た。
【００５９】
¹H NMR (CDC1₃) : 1.28 (s, 9H, 3 Me), 1.31 (s, 3H, Me), 1.33 (s, 3H, Me), 1.73 (s, 3H, Me), 1.90 (s, 3H, Me), 1.90 (m, 1H of C6'-H), 2.25 (s, 3H, Me), 2.55 (m, 1H of C6-H), 2.57 (s, 3H, Me), 3.736 (d, 1H of C3-H, J = 7.6 Hz), 4.26 (q, 2H of C20, J = 8.8 Hz), 4.38 (m, 1H of C7-H, J_l = 10.8 Hz, J₂ = 6.4 Hz), 4.82 (m, 1H, C3'-H), 4.86 (d, 1H of C14-H, J = 6.8 Hz), 4.90 (m, 1H of C5-H, J₁ = 2.3 Hz, J₂ = 9.9 Hz), 5.24 (d, 1H of NH), 6.10 (d, 1H of C2-H), 6.26 (s, 1H of C10-H), 6.47 (d, 1H of C13-H), 7.48-7.54 (m, 2H, arom), 7.58-7.64 (m, 1H, arom), 8.08-8.12 (m, 2H, arom)。
【００６０】
実施例６
（２′Ｒ，３′Ｒ）−１３−［Ｎ−Ｂｏｃ−２−メチル−３−（２−チエニル）イソセリノイル］−１４β−ヒドロキシバッカチンIII １，１４−カルボナート
７−ＴＥＳ−１４β−ヒドロキシバッカチンIII １，１４−カルボナート（０．２４g、０．３７５mmol）を、実施例３ｃに記載の条件で、（３Ｒ，４Ｓ）−１−tert−ブトキシカルボニル−３−トリエチルシリルオキシ−４−（チエン−２−イル）−３−メチル−アゼチジノン（０．３６g、０．９３６mmol）とカップリングさせた。ＨＦ／ピリジン溶液による脱保護後、表題化合物を白色の固体（０．１８９g、２．１mmol、５５％）を得た。
【００６１】
¹H NMR (CDC1₂) : δ 1.25 (s, 9H, 3 Me), 1.28 (s, 3H, Me), 1.43 (s, 3H, Me), 1.75 (s, 3H, Me), 1.86 (s, 3H, Me), 1.92 (m, 1H of C6'-H, J_6-6' = 14.3 Hz), 2.24 (s, 3H, Me), 2.55 (m, 1H of C6-H), 2.63 (s, 3H, Me), 3.73 (d, 1H of C3-H, J = 8.0 Hz), 4.26 (q, 2H of C20, J = 8.5 Hz), 4.38 (m, 1H of C7-H, J_l = 10.8 Hz, J₂ = 7.0 Hz), 4.86 (d, 1H of C14-H, J = 7.0 Hz), 4.93 (m, 1H of C5-H, J₁ = 1.8 Hz, J₂ = 9.8 Hz), 5.26 (d, 1H, 1H of C3'-H, J = 9.5 Hz), 5.44 (d, 1H of NH), 6.12 (d, 1H of C2-H), 6.28 (s, 1H of C10-H), 7.07 (dd, 5.0, 3.6 Hz, H-3 チエニル), 7.16 (dd, 3.6, 1.0, H-4 チエニル), 6.47 (d, 1H of C13-H), 7.35 (dd, 5.0, 1.0, H-5 チエニル), 7.44-7.50 (m, 2H, arom), 7.58-7.62 (m, 1H, arom), 8.02-8.08 (m, 2H, arom)。
【００６２】
薬理試験
薬理試験を、ジメチルスルホキシド中、０．１％濃度の本発明の化合物で実施した。Ａ２７８０ｗｔ、Ａ２７８０ｃｉｓ、Ａ２７８０ａｄｒ及びＡ２７８０ｔａｘ細胞株を使用した。細胞を、９６ウェル平底プレート（Viewplates、パッカード）に培養した。２４時間後、培地を取り替え、そして洗浄後、試験化合物を含む培地を加えた。０．０１から始め、１０００００，０００ｎＭまでの、用量−反応対数曲線を、各々のプレートに対し４通り設けた。各々のアッセイを、２通り、３回行った。試験化合物の存在下で７２時間培養後、細胞を回収し、生存細胞数を、ＡＴＰｌｉｔｅキット（パッカード、Meridien、ミズーリ州、米国）及びＴｏｐｃｏｕｎｔ（パッカード）オートルミノメーターを用いて、ＡＴＰ用量により評価した。各々の薬物／細胞株について、用量−反応曲線をプロットし、予測される効果の二乗の逆数によって加重された、非線形回帰を使用したＳ字状−Ｅｍａｘモデル（sigmoid-Emax model）による３回の試験で得られた濃度−効果曲線から、ＩＣ₅₀値を計算した。７２時間の試験化合物に対する連続的な曝露後の得られたＩＣ₅₀値を、以下の表に報告する。
【００６３】
【表１】

【Technical field】
[0001]
The present invention relates to a novel taxane derivative having antitumor activity and a method for producing the same.
[0002]
More particularly, the present invention provides compounds of general formula (I):
[0003]
[Chemical Formula 3]

[0004]
[Where,
R is trifluoromethyl, phenyl, 2-furyl, 2-thienyl;
R ¹ is t-butoxycarbonyl or benzoyl;
R ₂ is hydroxy;
R ₃ is hydrogen or, together with R ₂ , the formula:
[0005]
[Formula 4]

[0006]
And provided that when R ₃ is hydrogen, R is not phenyl.]
It is related with the compound shown by these.
[0007]
The compound of formula (I) is a known pharmaceutical product having antitumor activity, paclitaxel or a derivative of docetaxel.
[0008]
Compounds of formula (I) wherein R ₃ is hydrogen and R is phenyl are described by Greene et al. In J. Chem. Soc. Perkin Trans.l, 1995, pp 1811-1815. Paclitaxel derivatives having a C-2'-methylated (2R, 3S) side chain are described by Kant et al. In Tetrahedron, vol. 37, n. 36, pp 6495-6498, 1996.
[0009]
Kant compares this structural modification with paclitaxel, the parent compound, in certain tests (inhibition of microtubule depolymerization) because of its better binding affinity for microtubules and better cytotoxicity for KBVI. Have shown significant enhancement of response. In addition, some of these taxanes synthesized from 14β-hydroxybaccatin 1,14-carbonate had improved water solubility. Finally, to inhibit the metabolic pathways associated with cytochrome P-450 class enzymes, one of these taxanes contains a trifluoromethyl substituent at the C-3 position. The improved pharmacological characteristics of these novel compounds will correlate well with modification of the activity spectrum for various tumor types.
[0010]
Preferred compounds of the invention are the following:
(2'R, 3'R) -13- [N-benzoyl-3- (2-furyl) -2-methyl-isoserinoyl] -baccatin III;
(2'R, 3'R) -13- [Nt-butoxycarbonyl-3- (2-furyl) -2-methyl-isoserinoyl] -baccatin III;
(2′R, 3 ′S) -13- [Nt-butoxycarbonyl-3-phenyl-2-methyl-isoserinoyl] -14β-hydroxybaccatin III 1,14-carbonate;
(2′R, 3′R) -13- [Nt-butoxycarbonyl-3-trifluoromethyl-2-methyl-isoserinoyl] -14β-hydroxybaccatin III 1,14-carbonate;
(2′R, 3′R) -13- [Nt-butoxycarbonyl-3- (2-furyl) -2-methyl-isoserinoyl] -14β-hydroxybaccatin III 1,14-carbonate;
(2'R, 3'R) -13- [Nt-Butoxycarbonyl-3- (2-thienyl) -2-methyl-isoserinoyl] -14-hydroxybaccatin III 1,14-carbonate.
[0011]
The compound of formula (I) is represented by formula (II):
[0012]
[Chemical formula 5]

[0013]
Wherein R ₂ and R ₃ are as defined above and R ₄ is a protecting group, preferably triethylsilyl.
A compound of formula (III):
[0014]
[Chemical 6]

[0015]
(Wherein R and R ₁ are as defined above)
It is manufactured by reaction with the compound shown by these.
[0016]
The compounds of formula (III) are obtained from the corresponding alkyl esters by hydrolysis in alcohol solvents, in particular Tetrahedron Asymm. 2001, 12, 1015-1027 and J. Chem. Soc. Perkin Trans.l 1995, 1811-1816. Can be produced from a known methyl ester. The resulting acid is directly isolated from the baccatin III derivative of formula (II) in the presence of a suitable condensing agent such as di-2-pyridyl thionocarbonate and dimethylaminopyridine in a suitable solvent without isolation. Can be condensed. The hydroxy protecting group at the 7-position is removed to obtain the desired compound of formula (I).
[0017]
Alternatively, the compound of formula (II) is converted to formula (IV):
[0018]
[Chemical 7]

[0019]
(Wherein R is as defined above)
You may make it react with the compound shown by these.
[0020]
Compounds of formula (IV) in which R is phenyl are described in J. Org. Chem. 1991, 56, 1681-1684; Tetrahedron, 1992, 48, 6585-7012; EP 400971; US 5,175,315.
[0021]
Compounds of formula (IV) where R is trifluoromethyl can be prepared according to the following scheme.
[0022]
[Chemical 8]

[0023]
A compound of formula (IV) in which R is 2-furyl is known from (3R, 4R) -4- (furan-2-yl) -3, known from J. Org. Chem. 1999, 64, 4643-4651. It can be prepared according to the following scheme starting from -hydroxy-3-methyl-azetidin-2-one.
[0024]
[Chemical 9]

[0025]
Compounds of formula (II) are known from J. Med. Chem. 1997, 40, 267-278.
[0026]
The reaction between compound (II) and compound (IV) is carried out in an aprotic solvent under an inert atmosphere.
[0027]
Typically, this reaction is carried out at a temperature of about -40 ° C to 5 ° C in the presence of sodium hexamethyldisilazane. The 7-position hydroxy protecting group is then removed to give the compound of formula (I).
[0028]
The compounds of the present invention are potent against breast, lung, ovarian, colon, prostate, kidney, pancreatic tumor cells and against cells resistant to known anti-tumor agents such as adriamycin, vinblastine and platinum derivatives. Antitumor activity.
[0029]
The present invention therefore relates to pharmaceutical formulations comprising an effective amount of a compound of the invention together with pharmacologically acceptable carriers and excipients. More particularly, the compounds may be prepared in the form of tablets, powders, granules, capsules, injection solutions, suppositories, emulsions, dispersions and the like. For intravenous administration, mixtures of liposomal preparations prepared with Chremophor L and ethanol, polysorbate and ethanol, or natural or synthetic phosphatidylcholine, or natural phospholipids in the presence of cholesterol are preferably used; For this purpose, soft gelatin capsules are preferably prepared so that the product is solubilized with polysorbate, PEG or mixtures thereof, optionally in the presence of phospholipids. Compound (I), at concentrations ranging from 50 to 500 mg / m ^2, may be administered to a human.
[0030]
The following examples illustrate the invention in more detail.
[0031]
Abbreviations used are as follows:
TES = triethylsilyl; DMF = dimethylformamide; DMAP = (N, N-dimethylamino) pyridine; NaHMDS = sodium hexamethyldisilazide; LiHMDS = lithium hexamethyldisilazide; THF = tetrahydrofuran; HMPA = hexamethylphosphoric acid Triamide.
[0032]
Example 1
(4S, 5R) -3-benzoyl-2- (2,4-dimethoxyphenyl) -4- (furan-2-yl) -5-methyloxazolidine-5-carboxylic acid methyl ester (0.128 g, 0.285 mmol) was stirred in K ₂ CO ₃ ( ₂ eq) and anhydrous medium at 25 ° C. for 24 hours. The reaction mixture was concentrated under reduced pressure, diluted with saturated NH ₄ Cl solution and extracted with ethyl acetate. The aqueous phase was acidified with 5% NaHSO ₄ to pH 4 and extracted with ethyl acetate. The organic phase was dried and the solvent was evaporated off. The residue (1 eq) was purified in the presence of di-2-pyridyl thionocarbonate (1 eq) and dimethylaminopyridine (0.5 eq) at 20 ° C. under argon atmosphere in the presence of 7-TES-baccatin III (0.04 g, 0 0.057 mol) in toluene solution. The reaction mixture was heated at 60 ° C. with stirring for 80 hours. After the addition of ethyl acetate, the reaction mixture was extracted with brine. The organic phase was washed with brine, then dried and evaporated. Chromatography (SiO _2, ethyl acetate / n-hexane, 1: 1), (2'R, 3'R) -13- [ N- benzoyl -N, O-(2,4-dimethoxy benzylidene) - 3- (2-Furanyl) -methylisoserinoyl] -7-TES-baccatin III (0.023 g, 2.28 mmol, 40%) was obtained.
[0033]
This compound has the following properties:
¹ H NMR (CDC1 ₃ ): δ = 0.61 (m, 6H, 3 CH ₂ ), 0.95 (t, 9H, 3 Me), 1.22 (s, 3H, Me), 1.27 (s, 3H, Me), 1.68 (s, 3H, Me), 1.89 (m, 1H of C6-H), 2.10-2.25 (m, 2H of C14-H), 2.21 (s, 3H, Me), 2.24 (s, 3H, Me), 2.33 (s, 3H, Me), 2.50 (m, 1H of C6-H), 2.65 (s, 3H, Me, OAc of C-4), 3.90-4.0 (m, 8H, 6H of 2 Me, 1H of C3-H, and 1H of OH), 4.12 (d, 1H of C20-H, J = 8.0 Hz), 4.32 (d, 1H, C20-H), 4.58 (m, 1H of C7-H, J _l = 5.8 Hz, J ₂ = 10.2 Hz), 4.95 (dd, 1H of C5-H, J ₁ = 1.5 Hz, J ₂ = 9.6 Hz), 5.55 (b, 1H of C3'-H), 5.70 (d, 1H of C2-H, J = 5.7 Hz), 6.30-6.40 (m, 2H), 6.40-6. 50 (m, 3H), 6.54 (s, 1H), 6.82 (s, 1H), 7.20-7.40 (m , 5H, arom), 7.40-7.50 (m, 3H, arom), 7.54-7.60 (m, 1H, arom), 8.04-8.07 (m, 2H, arom).
[0034]
This compound (0.023 g, 2.28 mmol) was dissolved in CH ₂ C1 ₂ (3.0 ml) and treated at 25 ° C. with a solution of acetyl chloride in methanol (70 μl acetyl chloride in 10 ml MeOH) (100 μl). . After 7 hours, the reaction mixture was extracted with water. The organic phase was dried and evaporated. The residue was chromatographed (SiO _2, hexane / ethyl acetate to n-, 1: 1) by separating, 0.019g (0.022mmol, 98%) of (2'R, 3'R) -3'- dephenyl -3 '-(2-furyl) -2'-methyl-paclitaxel was obtained.
[0035]
This compound has the following properties:
¹ H NMR (CDC1 ₃ ): δ = 1.13 (s, 3H, Me), 1.23 (s, 3H, Me), 1.55 (s, 3H, Me), 1.70 (s, 3H, Me), 1.81 (s, 3H, Me), 1.89 (m, 1H, H-6β), 2.07 (m, 1H of C14-H), 2.24 (s, 3H, Me), 2.40 (m, 1H of C14-H), 2.49 (d , 1H, OH of C7-H), 2.55 (m, 1H of C6-H), 2.65 (s, 3H, Me), 3.75 (s, 1H, OH), 3.82 (d, 1H of C3-H, J = 7.2 Hz), 4.21 (d, 1H of C20-H J = 8.4 Hz), 4.31 (d, 1H of C20-H), 4.40 (m, 1H of C7-H), 4.95 (dd, 1H of C5- H, J ₁ = 1.5 Hz, J ₂ = 9.6 Hz), 5.66 (d, 1H of C2-H), 5.79 (d, 1H, of C3'-H, J = 9.5 Hz), 6.26 (s, 1H of C10-H), 6.30 (m, 1H of C13-H), 6.41 (m, 2H, 2-furyl), 7.12 (d, 1H, NH), 7.33 (m, 2H, arom), 7.44 (m, 1H , 2-furyl), 7.46 (m, 1H, arom), 7.52 (m, 2H, arom), 7.60 (m, 1H, arom), 7.67 (m, 2H, arom), 8.19 (m, 2H, arom) .
[0036]
Example 2
Following the procedure described in Example 1, (4S, 5R) -N-Boc-N, O- (2,4-dimethoxybenzylidene) -3-phenyl-2-methylisocene methyl ester (J. Chem. Soc. Perkin Trans. 1995, 1811-1816) (0.154 g, 0.336 mmol) and 7-TES-baccatin III 1,14-carbonate ¹ (0.05 g, 0.067 mmol) to give 0.021 g ( 0.023 mmol, 34%) of (2′R, 3 ′S) -13- (N-Boc-2-methyl-3-phenylisoserinoyl) -14β-hydroxybaccatin III 1,14-carbonate is obtained. It was.
[0037]
This compound has the following properties:
¹ H NMR (CDC1 ₃ ): δ = 1.28 (s, 9H, 3 Me of t-Boc), 1.30 (s, 3H, Me of C-15), 1.38 (s, 3H, Me of C-15), 1.41 (s, 3H, Me of C2 '), 1.73 (s, 3H, Me of C-8), 1.84 (s, 3H, Me of C-12), 1.92 (m, 1H of C6-H), 2 , 26 (s, 3H, Me, OAc of C-10), 2.36 (b, 1H, OH), 2.56 (m, 1H of C6-H), 2.69 (s, 3H, Me, OAc of C-4) , 3.52-3.55 (b, 1H, OH), 3.73 (d, 1H of C3-H, J = 7.3 Hz), 4.25 (d, 1H of C20-H, J = 8.5 Hz), 4.31 (d, 1H, of C20-H), 4.38 (m, 1H of C7-H), 4.85 (d, 1H of C14-H, J = 7.1 Hz), 4.95 (dd, 1H of C5-H, J _l = 2.4 Hz, J ₂ = 9.6 Hz), 5.09 (d, 1H of C3'-H), 5.57 (d, 1H, NH, J = 10.0 Hz), 6.13 (d, 1H of C2-H), 6,27 (s, 1H of C10-H), 6.36 (m, 1H of C13-H), 7.30-7.40 (m, 5H, arom), 7.48 (m, 2H, arom), 7.59 (m, 1H, arom), 8.04 (m, 2H, arom).
[0038]
Example 3
a) (3R, 4S) -3-triethylsilyloxy-4- (furan-2-yl) -3-methyl-azetidinone triethylsilyl chloride (0.316 g, 2.1 mmol) and imidazole (0.100 g, 1. 5 mmol) at 25 ° C. under argon atmosphere at (3R, 4R) -4- (furan-2-yl) -3-hydroxy-3-methyl-azetidin-2-one in DMF (6.0 ml) 0.167 g, 1.0 mmol) was added to the stirred solution. The reaction solution was quenched after 4 hours with saturated aqueous NH ₄ Cl and extracted with ethyl acetate. The organic phase was dried and concentrated under reduced pressure. Chromatography of the residue (SiO _2, ethyl acetate / n-pentane, 1: 2), 0.190 g (0.67 mmol, 67%) of (3R, 4S) -4- (furan-2-yl) -3 -Triethylsilyloxy-3-methyl-azetidin-2-one was obtained.
[0039]
[α] _D ²⁰ = +42.8 (c 1.04, CHCl ₃ );
IR (CDCl ₃ , cm ^-1 ): 3600-3000, 3413, 2957, 1768, 1458, 1377, 1012;
¹ H NMR (CDCl ₃ ): δ = 0.65 (m, 6H, 3 CH ₂ ), 0.95 (t, 9H, 3 Me), 1.19 (s, 3H, Me), 4.54 (s, 1H), 6.27 (d , 1H, 2-furyl), 6.28 (m, 1H, 2-furyl), 6.72 (b, 1H, NH), 7.40 (m, 1H, 2-furyl);
¹³ H NMR (CDCl3): δ = 5.71 (CH ₂ ), 6.73 (Me), 19.3 (Me), 61.0 (CH), 89.1 (C), 108.1 (CH), 110.5 (CH), 142.7 (CH), 150.9 (C), 171.5 (C).
[0040]
b) (3R, 4S) -1-tert-butoxycarbonyl-3-triethylsilyloxy-4- (furan-2-yl) -3-methyl-azetidinone Derivative from the previous step (0.060 g, 0.21 mmol) , DMAP (0.010 g), and triethylamine (88 μl, 0.63 mmol) in CH ₂ Cl ₂ (3.0 ml) at 0 ° C. with di-tert-butyl dicarbonate (121 ml, 0.52 mmol) in CH 2. ₂ C1 ₂ (1.0 ml) solution was added. The reaction mixture was stirred at 25 ° C. for 1 hour and then quenched with saturated NH ₄ Cl. The mixture was extracted with ethyl acetate and the organic layer was washed with brine, dried and concentrated under reduced pressure. Chromatography (SiO _2, n-pentane / ethyl acetate, 8: 2), 0.077 g (0.020 mmol, 97%) of (3R, 4S) -1-tert- butoxycarbonyl-3-triethylsilyloxy - 4- (Furan-2-yl) -3-methyl-azetidinone was obtained.
[0041]
[α] _D ²⁰ = +26.6 (c 0.98, CHCl ₃ );
IR (CDCl ₃ , cm ^-1 ): 2958, 1813, 1726, 1327, 1152;
¹ H NMR (CDC1 ₃ ): δ = 0.54 (m, 6H, 3 CH ₂ ), 0.80 (t, 9H, 3 Me), 1.41 (s, 9H, 3 Me), 1.61 (s, 3H, Me), 4.73 (s, 1H), 6.27 (d, 1H, 2-furyl), 6.34 (m, 1H, 2-furyl), 7.37 (m, 1H, 2-furyl)
¹³ H NMR (CDC1 ₃ ): δ = 5.6 (CH ₂ ), 6.5 (Me), 23.0 (Me), 27.8 (3 Me), 62.7 (CH), 83.3 (C), 85.4 (C), 108.9 (CH ), 110.3 (CH), 142.4 (CH), 147.9 (C), 148.7 (C), 167.7 (C).
[0042]
c) (2'R, 3'R) -13- [N-Boc-2-methyl-3- (2-furanyl) -isoserinoyl] -baccatin III
Β-lactam from step b) (0.082 g, 0.214 mmol) and 7-TES-baccatin III (0.060 g, 0.086 mmol) were dissolved in THF under an argon atmosphere. The solution was cooled to -45 ° C. NaHMDS (1.0 M n-hexane solution, 2.5 eq) was added dropwise with stirring. The temperature was raised to -20 ° C in 4 hours. The reaction was quenched with saturated NH ₄ Cl and the mixture was extracted with ethyl acetate and dried. After evaporation of the solvent, the crude was purified by chromatography. The reaction product was dissolved in 1: 1 MeCN / pyridine solvent at 0 ° C. A solution of HF / pyridine (70/30) was added dropwise (0.1 ml / 10 mg reagent) dropwise. The reaction mixture was stirred at 0 ° C. for 1 hour, then at 25 ° C. for 6 hours and extracted with ethyl acetate. The extract was washed 3 times with saturated CuSO ₄ and water and dried over anhydrous MgSO ₄ to give 0.035 g (0.041 mmol, 48%) of the title product.
[0043]
¹ H NMR (CDC1 ₃ ): δ = 1.15 (s, 3H, Me a C-15), 1.23 (s, 9H, 3 Me of t-Boc), 1.30 (s, 3H, Me a C-15), 1.42 (s, 3H, Me a C2 '), 1.69 (s, 3H, Me a C-8), 1.84 (s, 3H, Me a C-12), 1.89 (m, 1H, H-6β), 2.15 (m, 1H, H-14), 2,25 (s, 3H, Me, OAc of C-10), 2.37 (m, 1H, H-14), 2.55 (m, 1H of C6-H), 2.61 (s, 3H, Me, OAc of C-4), 3.58-3.65 (b, 1H, OH), 3.82 (d, 1H of C3-H, J = 7.0 Hz), 4.18 (d, 1H of C20-H , J = 8.4 Hz), 4.31 (d, 1H of C20-H), 4.42 (dd, 1H of C7-H, J _l = 6.4 Hz, J ₂ = 10.8 Hz), 4.95 (dd, 1H of C5-H , J ₁ = 2.4 Hz, J ₂ = 9.6 Hz), 5.23 (d, 1H, J = 9.6 Hz, NH), 5.39 (d, 1H of C3'-H), 5.66 (d, 1H of C2-H) , 6.28 (s, 1H, of C10-H), 6.34 (m, 1H, 2-furyl), 6.36 (m, 1H, H-13), 6.38 (m, 1H, 2-furyl), 7.42 (m, 1H, 2-furyl), 7.49 (m, 2H, arom), 7.59 (m, 1H, arom), 8.14 (m, 2H, arom).
[0044]
Example 4
(2'R, 3'R) -13- [N-Boc-2-methyl-3- (2-furanyl) isoserinoyl] -14β-hydroxybaccatin III 1,14-carbonate 7-TES-14β-hydroxybacca Tin III 1,14-carbonate (0.080 g, 0.125 mmol) under the conditions described in Example 3c, (3R, 4S) -1-tert-butoxycarbonyl-3-triethylsilyloxy-4- (furan 2-yl) -3-methyl-azetidinone (0.12 g, 0.312 mmol) and coupled to (2′R, 3 ′S) -13- [N-Boc-2-methyl-3- (2 -Furanyl) isoserinoyl] -3 ', 7-diTES-14β-hydroxybaccatin III 1,14-carbonate (0.087 g, 0.0775 mmol, 62%).
[0045]
¹ H NMR (CDC1 ₃ ): δ = 0.5-0.6 (m, 12H, 6 CH ₂ ), 0.85-0.95 (m, 18H, 6 Me), 1.25 (s, 3H, Me), 1.29 (s, 9H, 3 Me), 1.31 (s, 3H, Me), 1.56 (s, 3H, Me), 1.75 (s, 3H, Me), 1.92 (m, 1H of C6-H, J _6-6 , = 14.3 Hz) , 2.02 (s, 3H, Me), 2.20 (s, 3H, Me), 2.51 (m, 1H of C6-H), 2.71 (s, 3H, Me), 3.76 (d, 1H of C3-H, J = 8.1 Hz), 4.26 (q, 2H of C20, J = 8.5 Hz), 4.44 (m, 1H of C7-H, J _l = 10.8 Hz, J ₂ = 7.0 Hz), 4.85 (d, 1H of C14-H, J = 7.0 Hz), 4.89 (m, 1H of C5-H, J ₁ = 1.8 Hz, J ₂ = 9.8 Hz), 5.24 (d, 1H, 1H of C3'-H, J = 10.0 Hz), 5.28 (d, 1H of NH), 6.10 (d, 1H of C2-H), 6.26 (d, 1H of 2-furyl), 6.38 (m, 1H of 2-furyl), 6.42 (s, 1H of C10-H), 6.47 (d, 1H of C13-H), 7.38 (m, 1H of 2-furyl), 7.42-7.50 (m, 2H, arom), 7.54 -7.60 (m, 1H, arom), 8.02-8.08 (m, 2H, arom).
[0046]
Such a derivative was treated with HF / pyridine solution to give the title compound (0.063 g, 0.70 mmol, 91%).
[0047]
¹ H NMR (CDC1 ₂ ): δ1.25 (s, 9H, 3 Me), 1.28 (s, 3H, Me), 1.43 (s, 3H, Me), 1.75 (s, 3H, Me), 1.86 (s , 3H, Me), 1.92 (m, 1H of C6'-H, J _{6-6 '} = 14.3 Hz), 2.24 (s, 3H, Me), 2.55 (m, 1H of C6-H), 2.63 (s , 3H, Me), 3.73 (d, 1H of C3-H, J = 8.0 Hz), 4.26 (q, 2H of C20, J = 8.5 Hz), 4.38 (m, 1H of C7-H, J ₁ = 10.8 Hz, J ₂ = 7.0 Hz), 4.86 (d, 1H of C14-H, J = 7.0 Hz), 4.93 (m, 1H of C5-H, J _l = 1.8 Hz, J ₂ = 9.8 Hz), 5.26 ( d, 1H, 1H of C3'-H, J = 9.5 Hz), 5.44 (d, 1H of NH), 6.12 (d, 1H of C2-H), 6.28 (s, 1H of C10-H), 6.36 ( d, 1H of 2-furyl), 6.40 (m, 1H of 2-furyl), 6.47 (d, 1H of C13-H), 7.42 (m, 1H of 2-furyl), 7.44-7.50 (m, 2H, arom), 7.58-7.62 (m, 1H, arom), 8.02-8.08 (m, 2H, arom).
[0048]
Example 5
a) (3R, 4R) -3-hydroxy-1- (4-methoxyphenyl) -3-methyl-4-trifluoromethyl-azetidin-2-one (J. Org. Chem., 1999, 64, 4643- (2651) (2S, 5S) -2-tert-butyl-2,5-dimethyl- [1,3] dioxolan-4-one (0.344 g, 2.03 mmol) in a THF solution. Of 1M in THF (2.4 ml, 2.4 mmol) was added at -78 ° C. After 30 minutes, a solution of HMPA and N- (4-methoxyphenyl) trifluoroacetal dimine (0.81 g, 4.00 mmol) was added successively (THF: HMPA = 85: 15). After 4 hours, the reaction mixture was treated with 5 ml of 1M aqueous CH ₃ COOH at −78 ° C. and extracted with 1N HCl, then NH ₄ Cl, and finally with brine. The organic phase was dried (Na ₂ SO ₄ ) and the solvent was removed by evaporation under reduced pressure. Chromatography of the residue (SiO _2, ethyl acetate / n-pentane, 1: 2) gave notation β- lactam (3R, 4R) and 0.25g (0.91mmol, 45%).
[0049]
[α] _D ²⁰ = +28.4 (c 1.01, CHCl ₃ );
¹ H NMR (CDC1 ₃ ): δ = 1.70 (d, 3H, Me, J = 1.2 Hz), 3.02-3.10 (b, 1H, OH), 3.78 (s, 3H, OMe), 4.33 (q, 1H, J _HF = 5.7 Hz), 6.85-7.40 (m, 4H, arom);
¹³ H NMR (CDC1 ₃ ): δ = 22.2 (Me), 55.5 (OMe), 63.9 (q, CH, J = 31 Hz), 82.8 (C), 114.4 (2 CH), 119.7 (2 CH), 123.7 (CF ₃ , J = 279 Hz), 129.3 (C), 157.3 (C), 167.9 (C).
[0050]
b) Compound (0.25 g, 0) from (3R, 4R) -3-triethylsilyloxy-1- (4-methoxy-phenyl) -3-methyl-4-trifluoromethyl-azetidin-2-one a) .91 mmol) in DMF (3.0 ml) at 25 ° C. were added Et ₃ SiCl (0.31 g, 2.0 mmol) and N-methylimidazole (0.28 g, 4 mmol). After stirring for 2 hours, the reaction mixture was poured into ice water, extracted with ethyl acetate and dried. The solvent was removed by evaporation and the residue was chromatographed to give 0.32 g (0.82 mmol, 90%) of the title product.
[0051]
IR (CDCl ₃ , cm ^-1 ): 2957, 2878, 1778, 1514, 1298, 1251;
¹ H NMR (CDC1 ₃ ): δ = 0.75 (m, 6H, 3 CH ₂ ), 0.98 (t, 9H, 3 Me), 1.65 (s, 3H, Me), 3.79 (s, 3H, OMe), 4.22 (q, 1H, J _HF = 5.9 Hz), 6.85-7.40 (m, 4H, arom);
¹³ H NMR (CDC1 ₃ ): δ = 5.7 (CH ₂ ), 6.6 (Me), 23.4 (Me), 55.5 (OMe), 64.3 (q, CH, J = 32 Hz), 84.0 (C), 114.4 ( 2 CH), 119.5 (2 CH), 123.6 (CF ₃ , J = 283 Hz), 129.8 (C), 157.1 (C), 167.1 (C).
[0052]
c) Compound (0.30 g, 0.77 mmol) from acetonitrile (13.0 ml) from (3R, 4R) -3-triethylsilyloxy-3-methyl-4-trifluoromethyl-azetidin-2-one b) To the solution was added cerium (IV) ammonium nitrate (1.5 g, 2.74 mmol) in 20.0 ml of water and 30 ml of water dropwise at -50 ° C. for 2 hours. The mixture was diluted with 30 ml of water and extracted with ethyl acetate. The organic phase was washed with saturated NaHCO ₃ , saturated NaHSO ₃ and again with saturated NaHCO ₃ . The organic layer was dried and concentrated under reduced pressure. The residue was purified by column chromatography _{(SiO 2, CH 2 Cl 2} / ethyl acetate, 3: 1) to give to give the title compound (0.168g, 0.59mmol, 77%) .
[0053]
¹ H NMR (CDC1 ₃ ): δ = 0.69 (m, 6H, 3 CH ₂ ), 0.94 (t, 9H, 3 Me), 1.60 (s, 3H, Me), 3.77 (q, 1H, J _HF = 6.2 Hz), 6.20-6.45 (b, 1H, NH);
¹³ H NMR (CDC1 ₃ ): δ = 5.8 (CH ₂ ), 6.8 (Me), 23.7 (Me), 61.0 (q, CH, J = 32 Hz), 86.3 (C), 123.8 (CF ₃ , J = 280 Hz), 170.8 (C).
[0054]
d) Compound from (3R, 4R) -1- (tert-butoxycarbonyl) -3-triethylsilyloxy-3-methyl-trifluoromethyl-azetidin-2-one c) (0.168 g, 0.59 mmol) , DMAP (10 mg) and triethylamine (0.25 ml, 1.77 mmol) in dichloromethane (2.0 ml) at 0 ° C. with di-tert-butyl dicarbonate (0.32 g, 1.47 mmol) in dichloromethane (1 0.0 ml) solution was added. The reaction mixture was stirred at 25 ° C. for 2 hours and then quenched with saturated NH ₄ Cl. The mixture was extracted with ethyl acetate and the organic layer was washed with brine, dried and concentrated under reduced pressure. Chromatography (SiO _2, n-pentane / ethyl acetate, 4: 1) to give the title product 0.210 g (0.56 mmol, 95%) by.
[0055]
¹ H NMR (CDC1 ₃ ): δ = 0.69 (m, 6H, 3 CH ₂ ), 0.95 (t, 9H, 3 Me), 1.52 (s, 9H, 3 Me), 1.63 (s, 3H, Me), 4.10 (q, 1H, J _HF = 6.2 Hz);
¹³ H NMR (CDC1 ₃ ): δ = 5.6 (CH ₂ ), 6.5 (Me), 23.5 (Me), 27.8 (3 Me), 63.2 (q, CH, J = 32 Hz), 84.7 (C), 122.8 (CF ₃ , J = 281 Hz), 147.2 (C), 167.4 (C).
[0056]
e) (2′R, 3′R) -13- [N-Boc-2-methyl-3-trifluoromethylisoserinoyl] -14β-hydroxybaccatin III 1,14-carbonate According to the procedure of Example 4. 7-TES-14β-hydroxybaccatin III (0.080 g, 0.11 mmol) with the compound from d) (0.103 g, 0.27 mmol) to give 7,3′-di- -TES-derivative (0.071 g, 0.063 mmol, 57%) was obtained.
[0057]
¹ H NMR (CDC1 ₃ ): δ = 0.62-0.72 (m, 12H, 6 CH ₂ ), 0.88-0.95 (m, 18H, 6 Me), 1.25 (s, 9H, 3 Me), 1.31 (s, 3H , Me), 1.52 (s, 6H, 2 Me), 1.63 (s, 3H, Me), 1.90 (m, 1H of C6'-H), 2.00 (s, 3H, Me), 2.19 (s, 3H, Me), 2.48 (m, 1H of C6-H), 2.58 (s, 3H, Me), 3.74 (d, 1H of C3-H, J = 7.5 Hz), 4.24 (q, 2H of C20, J = 8.8 Hz), 4.41 (m, 1H of C7-H, J _l = 10.6 Hz, J ₂ = 6.4 Hz), 4.74 (m, 1H, C3'-H), 4.83 (d, 1H of C14-H, J = 6.9 Hz), 4.86 (m, 1H of C5-H, J _l = 1.90Hz, J ₂ = 9.8 Hz), 5.10 (d, 1H of NH), 6.10 (d, 1H of C2-H), 6.40 (s , 1H of C10-H), 6.42 (m, 1H of C13-H), 7.44-7.50 (m, 2H, arom), 7.58-7.60 (m, 1H, arom), 8.06-8.10 (m, 2H, arom ).
[0058]
This derivative was treated with a HF / pyridine solution to give the title compound (0.048 g, 0.54 mmol, 85%).
[0059]
¹ H NMR (CDC1 ₃ ): 1.28 (s, 9H, 3 Me), 1.31 (s, 3H, Me), 1.33 (s, 3H, Me), 1.73 (s, 3H, Me), 1.90 (s, 3H , Me), 1.90 (m, 1H of C6'-H), 2.25 (s, 3H, Me), 2.55 (m, 1H of C6-H), 2.57 (s, 3H, Me), 3.736 (d, 1H of C3-H, J = 7.6 Hz), 4.26 (q, 2H of C20, J = 8.8 Hz), 4.38 (m, 1H of C7-H, J _l = 10.8 Hz, J ₂ = 6.4 Hz), 4.82 ( m, 1H, C3'-H), 4.86 (d, 1H of C14-H, J = 6.8 Hz), 4.90 (m, 1H of C5-H, J ₁ = 2.3 Hz, J ₂ = 9.9 Hz), 5.24 (d, 1H of NH), 6.10 (d, 1H of C2-H), 6.26 (s, 1H of C10-H), 6.47 (d, 1H of C13-H), 7.48-7.54 (m, 2H, arom ), 7.58-7.64 (m, 1H, arom), 8.08-8.12 (m, 2H, arom).
[0060]
Example 6
(2′R, 3′R) -13- [N-Boc-2-methyl-3- (2-thienyl) isoserinoyl] -14β-hydroxybaccatin III 1,14-carbonate 7-TES-14β-hydroxybacca Tin III 1,14-carbonate (0.24 g, 0.375 mmol) was prepared under the conditions described in Example 3c (3R, 4S) -1-tert-butoxycarbonyl-3-triethylsilyloxy-4- (thien Coupled with 2-yl) -3-methyl-azetidinone (0.36 g, 0.936 mmol). After deprotection with HF / pyridine solution, the title compound was obtained as a white solid (0.189 g, 2.1 mmol, 55%).
[0061]
¹ H NMR (CDC1 ₂ ): δ 1.25 (s, 9H, 3 Me), 1.28 (s, 3H, Me), 1.43 (s, 3H, Me), 1.75 (s, 3H, Me), 1.86 (s, 3H, Me), 1.92 (m, 1H of C6'-H, J _{6-6 '} = 14.3 Hz), 2.24 (s, 3H, Me), 2.55 (m, 1H of C6-H), 2.63 (s, 3H, Me), 3.73 (d, 1H of C3-H, J = 8.0 Hz), 4.26 (q, 2H of C20, J = 8.5 Hz), 4.38 (m, 1H of C7-H, J _l = 10.8 Hz , J ₂ = 7.0 Hz), 4.86 (d, 1H of C14-H, J = 7.0 Hz), 4.93 (m, 1H of C5-H, J ₁ = 1.8 Hz, J ₂ = 9.8 Hz), 5.26 (d , 1H, 1H of C3'-H, J = 9.5 Hz), 5.44 (d, 1H of NH), 6.12 (d, 1H of C2-H), 6.28 (s, 1H of C10-H), 7.07 (dd , 5.0, 3.6 Hz, H-3 thienyl), 7.16 (dd, 3.6, 1.0, H-4 thienyl), 6.47 (d, 1H of C13-H), 7.35 (dd, 5.0, 1.0, H-5 thienyl) 7.44-7.50 (m, 2H, arom), 7.58-7.62 (m, 1H, arom), 8.02-8.08 (m, 2H, arom).
[0062]
Pharmacological tests Pharmacological tests were carried out with the compounds of the invention at a concentration of 0.1% in dimethyl sulfoxide. A2780 wt, A2780cis, A2780adr and A2780tax cell lines were used. Cells were cultured in 96 well flat bottom plates (Viewplates, Packard). After 24 hours, the medium was changed and after washing, medium containing the test compound was added. There were four dose-response logarithmic curves for each plate starting at 0.01 and going up to 100,000,000 nM. Each assay was performed in duplicate and in triplicate. After 72 hours of culture in the presence of the test compound, cells were harvested and viable cell number was assessed by ATP dose using ATPlite kit (Packard, Meridien, MO, USA) and Topcount (Packard) autoluminometer. . For each drug / cell line, a dose-response curve is plotted, and weighted by the inverse of the square of the expected effect, three times by sigmoid-Emax model using non-linear regression. IC ₅₀ values were calculated from the concentration-effect curves obtained in the test. The IC ₅₀ values obtained after continuous exposure to the test compound for 72 hours are reported in the table below.
[0063]
[Table 1]

Claims (4)

Formula (I):

[Where,
R is trifluoromethyl, phenyl, 2-furyl, 2-thienyl;
R ¹ is t-butoxycarbonyl or benzoyl;
R ₂ is hydroxy;
R ₃ is hydrogen or, together with R ₂ , the formula:

And provided that when R ₃ is hydrogen, R is not phenyl.]
A compound represented by As a compound of formula (I):
(2'R, 3'R) -13- [N-benzoyl-3- (2-furyl) -2-methyl-isoserinoyl] -baccatin III;
(2'R, 3'R) -13- [Nt-butoxycarbonyl-3- (2-furyl) -2-methyl-isoserinoyl] -baccatin III;
(2'R, 3'S) -13- [Nt-butoxycarbonyl-3-phenyl-2-methyl-isoserinoyl] -14-hydroxybaccatin III 1,14-carbonate;
(2'R, 3'R) -13- [Nt-butoxycarbonyl-3-trifluoromethyl-2-methyl-isoserinoyl] -14-hydroxybaccatin III 1,14-carbonate;
(2'R, 3'R) -13- [Nt-butoxycarbonyl-3- (2-furyl) -2-methyl-isoserinoyl] -14-hydroxybaccatin III 1,14-carbonate;
(2'R, 3'R) -13- [Nt-Butoxycarbonyl-3- (2-thienyl) -2-methyl-isoserinoyl] -14-hydroxybaccatin III 1,14-carbonate. A pharmaceutical composition comprising one of the compounds of formula (I) together with a pharmaceutically acceptable carrier and excipient. Use of a compound of formula (I) for the manufacture of a medicament having antitumor activity.