11-[(4-{2-[2(2-Aminoethoxy)ethoxy]ethyl}piperazine-1-yl)acetyl]-5,11-dihydro-6H-pyrido-[2,3-b][1,4]benzodiazepine-6-one | 745048-24-2

• 分子结构分类: 有机化合物 - 有机杂环化合物 - 苯二氮卓类
• 英文名称: 11-[(4-{2-[2(2-Aminoethoxy)ethoxy]ethyl}piperazine-1-yl)acetyl]-5,11-dihydro-6H-pyrido-[2,3-b][1,4]benzodiazepine-6-one
• 英文别名: 11-[(4-{2-[2-(2-aminoethoxy)ethoxy]ethyl}piperazin-1-yl)acetyl]-5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepin-6-one；11-[2-[4-[2-[2-(2-aminoethoxy)ethoxy]ethyl]piperazin-1-yl]acetyl]-5H-pyrido[2,3-b][1,4]benzodiazepin-6-one
• CAS: 745048-24-2
• 化学式: C₂₄H₃₂N₆O₄
• 分子量: 468.556
• InChiKey: OFGKUZPFCKMULR-UHFFFAOYSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -1.1
• 重原子数：
  34
• 可旋转键数：
  10
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.46
• 拓扑面积：
  113
• 氢给体数：
  2
• 氢受体数：
  8

反应信息

• 作为反应物：
  描述：

  [4-((4-diethylaminophenyl)-{4-[5-(2,5-dioxopyrrolidin-1-yloxycarbonyl)pentylsulfamoyl]-2-sulfophenyl}methylene)cyclohexa-2,5-dienylidene]diethylammonium 、 11-[(4-{2-[2(2-Aminoethoxy)ethoxy]ethyl}piperazine-1-yl)acetyl]-5,11-dihydro-6H-pyrido-[2,3-b][1,4]benzodiazepine-6-one 在 N,N-二异丙基乙胺 、 N,N-二甲基甲酰胺 作用下， 以 乙腈 为溶剂， 反应 3.0h， 以34%的产率得到2-[[4-(diethylamino)phenyl]-(4-diethylazaniumylidenecyclohexa-2,5-dien-1-ylidene)methyl]-5-[[6-oxo-6-[2-[2-[2-[4-[2-oxo-2-(6-oxo-5H-pyrido[2,3-b][1,4]benzodiazepin-11-yl)ethyl]piperazin-1-yl]ethoxy]ethoxy]ethylamino]hexyl]sulfamoyl]benzenesulfonate
  参考文献：
  名称：

  On the Use of Nonfluorescent Dye Labeled Ligands in FRET-Based Receptor Binding Studies

  摘要：

  The efficiency of fluorescence resonance energy transfer (FRET) is dependent upon donor-acceptor proximity and spectral overlap, whether the acceptor partner is fluorescent or not. We report here on the design, synthesis, and characterization of two novel pirenzepine derivatives that were coupled to patent blue VF and pinacyanol dyes. These nonfluorescent compounds, when added to cells stably expressing enhanced green fluorescent protein (EGFP)fused muscarinic M1 receptors, promote EGFP fluorescence extinction in a time-, concentration-, and atropine-dependent manner. They display nanomolar affinity for the muscarinic receptor, determined using either FRET or classical radioligand binding conditions. We provide evidence that these compounds behave as potent acceptors of energy from excited EGFP with quenching efficiencies comparable to those of analogous fluorescent bodipy or rhodamine red pirenzepine derivatives. The advantages they offer over fluorescent ligands are illustrated and discussed in terms of reliability, sensitivity, and wider applicability of FRET-based receptor binding assays.

  DOI：

  10.1021/jm050459+
• 作为产物：
  描述：

  邻硝基苯甲酸 在 吡啶 、 盐酸 、 草酰氯 、 水 、 potassium carbonate 、 三乙胺 、 N,N-二甲基甲酰胺 、 三苯基膦 、 tin(ll) chloride 作用下， 以 四氢呋喃 、 1,4-二氧六环 、 乙腈 为溶剂， 反应 43.33h， 生成 11-[(4-{2-[2(2-Aminoethoxy)ethoxy]ethyl}piperazine-1-yl)acetyl]-5,11-dihydro-6H-pyrido-[2,3-b][1,4]benzodiazepine-6-one
  参考文献：
  名称：

  Fluorescent Pirenzepine Derivatives as Potential Bitopic Ligands of the Human M1 Muscarinic Receptor

  摘要：

  Following a recent description of fluorescence resonance energy transfer between enhanced green fluorescent protein (EGFP)-fused human muscarinic M1 receptors and Bodipy-labeled pirenzepine, we synthesized seven fluorescent derivatives of this antagonist in order to further characterize ligand-receptor interactions. These compounds carry Bodipy [558/568], Rhodamine Red-X [560/580], or Fluorolink Cy3 [550/570] fluorophores connected to pirenzepine through various linkers. All molecules reversibly bind with high affinity to M1 receptors (radioligand and energy transfer binding experiments) provided that the linker contains more than six atoms. The energy transfer efficiency exhibits modest variations among ligands, indicating that the distance separating EGFP from the fluorophores remains almost constant. This also supports the notion that the fluorophores may bind to the receptor protein. Kinetic analyses reveal that the dissociation of two Bodipy derivatives (10 or 12 atom long linkers) is sensitive to the presence of the allosteric modulator brucine, while that of all other molecules (15-24 atom long linkers) is not. The data favor the idea that these analogues might interact with both the acetylcholine and the brucine binding domains.

  DOI：

  10.1021/jm040800a

文献信息

• Method for Generating a Recombinant Clonal Cell Line and Novel Reagents for Use in the Method
  申请人：Hill Stephen John

  公开号：US20110300116A1

  公开（公告）日：2011-12-08

  A method for generating a recombinant clonal cell line expressing a target cell surface receptor at a specific level of expression from a cell population comprising cells transfected with a plasmid encoding the cDNA sequence of the target receptor and expressing the target cell surface receptor, the method comprising (c) incubating the cell population with a receptor specific fluorescent ligand (d) selecting single cells from step (c) expressing the target cell surface receptor by monitoring the specific binding of the fluorescent ligand using flow cytometry; and novel fluorescent ligands.

  一种用于从包含转染携带目标受体cDNA序列的质粒并表达目标细胞表面受体的细胞群体中生成表达特定水平目标细胞表面受体的重组克隆细胞系的方法，该方法包括：(c)将细胞群体与受体特异性荧光配体孵育；(d)通过使用流式细胞仪监测荧光配体的特异结合来选择步骤(c)中表达目标细胞表面受体的单个细胞；以及新型荧光配体。
• A METHOD FOR GENERATING A RECOMBINANT CLONAL CELL LINE AND NOVEL REAGENTS FOR USE IN THE METHOD
  申请人：Cellaura Technologies LTD

  公开号：EP2238248A2

  公开（公告）日：2010-10-13
• [EN] A METHOD FOR GENERATING A RECOMBINANT CLONAL CELL LINE AND NOVEL REAGENTS FOR USE IN THE METHOD<br/>[FR] MÉTHODE D'OBTENTION D'UNE LIGNÉE DE CELLULES CLONALES DE RECOMBINAISON ET NOUVEAUX RÉACTIFS UTILISÉS PAR LA MÉTHODE
  申请人：CELLAURA TECHNOLOGIES LTD

  公开号：WO2009040555A2

  公开（公告）日：2009-04-02

  A method for generating a recombinant clonal cell line expressing a target cell surface receptor at a specific level of expression from a cell population comprising cells transfected with a plasmid encoding the cDNA sequence of the target receptor and expressing the target cell surface receptor, the method comprising (c) incubating the cell population with a receptor specific fluorescent ligand (d) selecting single cells from step (c) expressing the target cell surface receptor by monitoring the specific binding of the fluorescent ligand using flow cytometry; and novel fluorescent ligands.
• Fluorescent Pirenzepine Derivatives as Potential Bitopic Ligands of the Human M1 Muscarinic Receptor
  作者：Chouaib Tahtaoui、Isabelle Parrot、Philippe Klotz、Fabrice Guillier、Jean-Luc Galzi、Marcel Hibert、Brigitte Ilien

  DOI：10.1021/jm040800a

  日期：2004.8.1

  Following a recent description of fluorescence resonance energy transfer between enhanced green fluorescent protein (EGFP)-fused human muscarinic M1 receptors and Bodipy-labeled pirenzepine, we synthesized seven fluorescent derivatives of this antagonist in order to further characterize ligand-receptor interactions. These compounds carry Bodipy [558/568], Rhodamine Red-X [560/580], or Fluorolink Cy3 [550/570] fluorophores connected to pirenzepine through various linkers. All molecules reversibly bind with high affinity to M1 receptors (radioligand and energy transfer binding experiments) provided that the linker contains more than six atoms. The energy transfer efficiency exhibits modest variations among ligands, indicating that the distance separating EGFP from the fluorophores remains almost constant. This also supports the notion that the fluorophores may bind to the receptor protein. Kinetic analyses reveal that the dissociation of two Bodipy derivatives (10 or 12 atom long linkers) is sensitive to the presence of the allosteric modulator brucine, while that of all other molecules (15-24 atom long linkers) is not. The data favor the idea that these analogues might interact with both the acetylcholine and the brucine binding domains.
• On the Use of Nonfluorescent Dye Labeled Ligands in FRET-Based Receptor Binding Studies
  作者：Chouaib Tahtaoui、Fabrice Guillier、Philippe Klotz、Jean-Luc Galzi、Marcel Hibert、Brigitte Ilien

  DOI：10.1021/jm050459+

  日期：2005.12.1

  The efficiency of fluorescence resonance energy transfer (FRET) is dependent upon donor-acceptor proximity and spectral overlap, whether the acceptor partner is fluorescent or not. We report here on the design, synthesis, and characterization of two novel pirenzepine derivatives that were coupled to patent blue VF and pinacyanol dyes. These nonfluorescent compounds, when added to cells stably expressing enhanced green fluorescent protein (EGFP)fused muscarinic M1 receptors, promote EGFP fluorescence extinction in a time-, concentration-, and atropine-dependent manner. They display nanomolar affinity for the muscarinic receptor, determined using either FRET or classical radioligand binding conditions. We provide evidence that these compounds behave as potent acceptors of energy from excited EGFP with quenching efficiencies comparable to those of analogous fluorescent bodipy or rhodamine red pirenzepine derivatives. The advantages they offer over fluorescent ligands are illustrated and discussed in terms of reliability, sensitivity, and wider applicability of FRET-based receptor binding assays.