1-(4-Hydroxyphenyl)-3-(4-methyl-2-oxochromen-7-yl)urea | 960522-81-0

• 分子结构分类: 有机化合物 - 苯丙烷和聚酮 - 香豆素及其衍生物
• 英文名称: 1-(4-Hydroxyphenyl)-3-(4-methyl-2-oxochromen-7-yl)urea
• CAS: 960522-81-0
• 化学式: C₁₇H₁₄N₂O₄
• 分子量: 310.309
• InChiKey: QALXYEVCEYPEGK-UHFFFAOYSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  2.5
• 重原子数：
  23
• 可旋转键数：
  2
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.06
• 拓扑面积：
  87.7
• 氢给体数：
  3
• 氢受体数：
  4

反应信息

• 作为产物：
  描述：

  7-氨基-4-甲基香豆素 、 4-Hydroxyphenylisocyanat 在 吡啶 作用下， 以 N,N-二甲基甲酰胺 、 乙腈 为溶剂， 反应 20.0h， 生成 1-(4-Hydroxyphenyl)-3-(4-methyl-2-oxochromen-7-yl)urea
  参考文献：
  名称：

  Discovery of small-molecule inhibitors of tyrosinase

  摘要：

  To identify novel inhibitors of tyrosinase, a fluorescent assay was developed which is suitable for high-throughput screening. In the assay, oxidation of the substrate by tyrosinase leads to the release of a fluorescent coumarin. Several small molecules were identified that inhibited mushroom tyrosinase in vitro and human tyrosinase in cell culture. These compounds may represent lead structures for therapies targeted at disorders of hyperpigmentation. (c) 2007 Elsevier Ltd. All rights reserved.

  DOI：

  10.1016/j.bmcl.2007.10.014

文献信息

• Means and methods for the determination of the biological activity of BoNT/E in cells
  申请人：MERZ PHARMA GMBH & CO. KGAA

  公开号：US10900955B2

  公开（公告）日：2021-01-26

  The present invention pertains to a polyclonal or monoclonal antibody specifically binding to BoNT/E-cleaved SNAP-25. Further, the invention provides a method for directly determining the biological activity of BoNT/E in cells, comprising the steps of: a) incubating cells susceptible to BoNT/E intoxication with BoNT/E for a time and under conditions which allow for the BoNT/E to exert its biological activity; b) fixing the cells and, optionally, permeabilizing the cells with a detergent; c) contacting the cells with at least a first capture antibody specifically binding to non-cleaved and BoNT/E-cleaved SNAP-25, and with at least a second capture antibody specifically binding to BoNT/E-cleaved SNAP-25, wherein the second capture antibody is an antibody of the invention, under conditions which allow for binding of said capture antibodies to the indicated substrates; d) contacting the cells with at least a first detection antibody specifically binding to the first capture antibody, under conditions which allow for binding of said first detection antibody to said first capture antibody, thus forming first detection complexes, and with at least a second detection antibody specifically binding to the second capture antibody, under conditions which allow for binding of said second detection antibody to said second capture antibody, thus forming second detection complexes, and wherein the first detection antibody is different from the second detection antibody; e) determining the amount of the first and second detection complexes of step d); and f) calculating the amount of SNAP-25 cleaved by BoNT/E in said cells by means of the second detection complexes, thereby determining the biological activity of BoNT/E in said cells. Furthermore, the invention relates to a kit for carrying out the method of the invention.

  本发明涉及一种特异性结合 BoNT/E 裂解 SNAP-25 的多克隆或单克隆抗体。此外，本发明还提供了一种直接测定细胞中 BoNT/E 生物活性的方法，包括以下步骤：a) 将易受 BoNT/E 感染的细胞与 BoNT/E 培养一段时间，并在允许 BoNT/E 发挥其生物活性的条件下培养；b) 固定细胞，可选地，用去垢剂使细胞渗透；c) 在允许所述捕获抗体与所述底物结合的条件下，使细胞与至少一种特异性结合非裂解和 BoNT/E 裂解 SNAP-25 的第一捕获抗体和至少一种特异性结合 BoNT/E 裂解 SNAP-25 的第二捕获抗体（其中第二捕获抗体是本发明的抗体）接触；d) 在允许所述第一检测抗体与所述第一捕获抗体结合从而形成第一检测复合物的条件下，至少用特异性结合第一捕获抗体的第一检测抗体与细胞接触，以及在允许所述第二检测抗体与所述第二捕获抗体结合从而形成第二检测复合物的条件下，至少用特异性结合第二捕获抗体的第二检测抗体与细胞接触，其中第一检测抗体不同于第二检测抗体；e) 确定步骤 d) 中第一和第二检测复合物的数量；以及 f) 通过第二检测复合物计算所述细胞中被 BoNT/E 裂解的 SNAP-25 的数量，从而确定所述细胞中 BoNT/E 的生物活性。此外，本发明还涉及一种用于实施本发明方法的试剂盒。
• MEANS AND METHODS FOR THE DETERMINATION OF THE BIOLOGICAL ACTIVITY OF NEUROTOXIN POLYPEPTIDES IN CELLS
  申请人：Merz Pharma GmbH & Co. KGaA

  公开号：EP3014267A1

  公开（公告）日：2016-05-04
• MEANS AND METHODS FOR THE DETERMINATION OF THE BIOLOGICAL ACTIVITY OF BONT/E IN CELLS
  申请人：Merz Pharma GmbH & Co. KGaA

  公开号：EP3233911A1

  公开（公告）日：2017-10-25
• Means and Methods for the determination of the biological activity of Neurotoxin polypeptides in cells
  申请人：MERZ PHARMA GMBH & CO. KGAA

  公开号：US20160202245A1

  公开（公告）日：2016-07-14

  The present invention pertains to a method for directly determining the biological activity of a Neurotoxin polypeptide in cells, comprising the steps of: a) incubating cells susceptible to Neurotoxin intoxication with a Neurotoxin polypeptide for a time and under conditions which allow for the Neurotoxin polypeptide to exert its biological activity; b) fixing the cells and, optionally, permeabilizing the cells with a detergent; c) contacting the cells with at least a first capture antibody specifically binding to the non-cleaved and Neurotoxin-cleaved substrate and with at least a second capture antibody specifically binding to the cleavage site of the Neurotoxin-cleaved substrate, under conditions which allow for binding of said capture antibodies to said substrates; d) contacting the cells with at least a first detection antibody specifically binding to the first capture antibody, under conditions which allow for binding of said first detection antibody to said first capture antibody, thus forming first detection complexes, and at least a second detection antibody specifically binding to the second capture antibody, under conditions which allow for binding of said second detection antibody to said second capture antibody, thus forming second detection complexes; e) determining the amount of the first and second detection complexes of step d), and f) calculating the amount of substrate cleaved by said Neurotoxin polypeptide in said cells by means of the second detection complexes, thereby determining the biological activity of said Neurotoxin polypeptide in said cells. The invention further provides for a kit for carrying out the method of the invention.
• [EN] MEANS AND METHODS FOR THE DETERMINATION OF THE BIOLOGICAL ACTIVITY OF NEUROTOXIN POLYPEPTIDES IN CELLS<br/>[FR] MOYENS ET PROCÉDÉS POUR DÉTERMINER L'ACTIVITÉ BIOLOGIQUE DE POLYPEPTIDES DE NEUROTOXINE DANS DES CELLULES
  申请人：MERZ PHARMA GMBH & CO KGAA

  公开号：WO2014207109A1

  公开（公告）日：2014-12-31

  The present invention pertains to methods and kits for determining the biological activity of Clostridial Neurotoxins. The method of the invention allows for the direct determination of the biological activity of a Neurotoxin polypeptide in cells. Cells susceptible to a Neurotoxin, e.g. neurological cells, are incubated with the neurotoxin (e.g. BoTN/ A), then fixed, and stained using an antibody specifically binding the cleaved neurotoxin (e.g. SNAP-25) and an antibody binding to both cleaved and uncleaved neurotoxin, for determination of total amount or content of neurotoxin substrate in the cells. The biological activity of said neurotoxin polypeptide is determined directly in the cells, and calculated by means of detection of the two complexes.