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    Luminescence-based methods and probes for measuring cytochrome P450 activity
    1.
    发明授权
    Luminescence-based methods and probes for measuring cytochrome P450 activity 有权
    用于测量细胞色素P450活性的基于发光的方法和探针

    公开(公告)号：US08106052B2

    公开(公告)日：2012-01-31

    申请号：US12754164

    申请日：2010-04-05

    申请人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel Kin Sang Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

    发明人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel Kin Sang Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

    IPC分类号： A61K31/403 ,  A61K31/4164 ,  A61K31/422 ,  A61K31/437 ,  A61K31/4985 ,  C07D413/04 ,  C07D417/04 ,  C07D471/04 ,  C07D487/04

    CPC分类号： G01N33/502 ,  A61K49/0008 ,  C07D417/04 ,  C12Q1/66 ,  G01N33/5088 ,  G01N33/582 ,  G01N2333/90245 ,  G01N2333/90251 ,  G01N2500/10

    摘要： The present invention provides methods, compositions, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferin or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. Upon addition of the luciferin derivative or other luminogenic molecule into a P450 reaction, the P450 enzyme metabolizes the molecule into a bioluminescent enzyme substrate, e.g., luciferin and/or luciferin derivative metabolite, in a P450 reaction. The resulting metabolite(s) serves as a substrate of the bioluminescent enzyme, e.g., luciferase, in a second light-generating reaction. Luminescent cytochrome P450 assays with low background signals and high sensitivity are disclosed and isoform selectivity is demonstrated. The present invention also provides an improved method for performing luciferase reactions which employs added pyrophosphatase to remove inorganic pyrophosphate, a luciferase inhibitor which may be present in the reaction mixture as a contaminant or may be generated during the reaction. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible luciferase inhibitors.

    摘要翻译： 本发明提供了可用于分析细胞，组织和动物中的代谢活性以及筛选测试化合物对细胞色素P450活性的影响的方法，组合物，底物和试剂盒。 特别地，使用发光分子的一步和两步法。 荧光素或coelenterazines，其是细胞色素P450底物并且也是生物发光酶，例如萤光素酶，原底物。 在P450反应中加入荧光素衍生物或其他发光分子时，P450酶在P450反应中将分子代谢为生物发光酶底物，例如荧光素和/或荧光素衍生物代谢产物。 所产生的代谢物在第二次发光反应中用作生物发光酶，例如荧光素酶的底物。 公开了具有低背景信号和高灵敏度的发光细胞色素P450测定，证明了同工型选择性。 本发明还提供一种用于进行荧光素酶反应的改进方法，其使用添加的焦磷酸酶来除去无机焦磷酸盐，荧光素酶抑制剂，其可作为污染物存在于反应混合物中，或可能在反应期间产生。 本发明方法还提供了一种使用荧光素酶稳定剂如可逆荧光素酶抑制剂在基于荧光素酶的测定中稳定和延长发光信号的方法。

    Luminescence-based methods and probes for measuring cytochrome P450 activity
    2.
    发明授权
    Luminescence-based methods and probes for measuring cytochrome P450 activity 有权
    用于测量细胞色素P450活性的基于发光的方法和探针

    公开(公告)号：US07692022B2

    公开(公告)日：2010-04-06

    申请号：US10665314

    申请日：2003-09-19

    申请人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel Kin Sang Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

    发明人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel Kin Sang Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

    IPC分类号： C07D277/68 ,  C07D417/04

    CPC分类号： G01N33/502 ,  A61K49/0008 ,  C07D417/04 ,  C12Q1/66 ,  G01N33/5088 ,  G01N33/582 ,  G01N2333/90245 ,  G01N2333/90251 ,  G01N2500/10

    摘要： The present invention provides methods, compositions, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferin or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. Upon addition of the luciferin derivative or other luminogenic molecule into a P450 reaction, the P450 enzyme metabolizes the molecule into a bioluminescent enzyme substrate, e.g., luciferin and/or luciferin derivative metabolite, in a P450 reaction. The resulting metabolite(s) serves as a substrate of the bioluminescent enzyme, e.g., luciferase, in a second light-generating reaction. Luminescent cytochrome P450 assays with low background signals and high sensitivity are disclosed and isoform selectivity is demonstrated. The present invention also provides an improved method for performing luciferase reactions which employs added pyrophosphatase to remove inorganic pyrophosphate, a luciferase inhibitor which may be present in the reaction mixture as a contaminant or may be generated during the reaction. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible luciferase inhibitors.

    摘要翻译： 本发明提供了可用于分析细胞，组织和动物中的代谢活性以及筛选测试化合物对细胞色素P450活性的影响的方法，组合物，底物和试剂盒。 特别地，使用发光分子的一步和两步法。 荧光素或coelenterazines，其是细胞色素P450底物并且也是生物发光酶，例如萤光素酶，原底物。 在P450反应中加入荧光素衍生物或其他发光分子时，P450酶在P450反应中将分子代谢为生物发光酶底物，例如荧光素和/或荧光素衍生物代谢产物。 所产生的代谢物在第二次发光反应中用作生物发光酶，例如荧光素酶的底物。 公开了具有低背景信号和高灵敏度的发光细胞色素P450测定，证明了同工型选择性。 本发明还提供一种用于进行荧光素酶反应的改进方法，其使用添加的焦磷酸酶来除去无机焦磷酸盐，荧光素酶抑制剂，其可作为污染物存在于反应混合物中，或可能在反应期间产生。 本发明方法还提供了一种使用荧光素酶稳定剂如可逆荧光素酶抑制剂在基于荧光素酶的测定中稳定和延长发光信号的方法。

    Bioluminescent protease assay with modified aminoluciferin or derivatives thereof
    5.
    发明授权
    Bioluminescent protease assay with modified aminoluciferin or derivatives thereof 有权
    具有修饰的氨基荧光素或其衍生物的生物发光蛋白酶测定

    公开(公告)号：US07384758B2

    公开(公告)日：2008-06-10

    申请号：US11346043

    申请日：2006-02-02

    申请人： Martha O'Brien ,  Keith V. Wood ,  Dieter Klaubert ,  William Daily

    发明人： Martha O'Brien ,  Keith V. Wood ,  Dieter Klaubert ,  William Daily

    IPC分类号： C12Q1/66

    CPC分类号： C07D277/66 ,  C12Q1/37 ,  C12Q1/66

    摘要： A sensitive bioluminescent assay to detect proteases, including caspases and proteases that specifically cleave a peptide substrate having aspartate, is provided. In one embodiment, the assay employs an aminoluciferin or a carboxy-terminal protected derivative thereof covalently linked via a peptide bond to a substrate for a caspase or an aminoluciferin or a carboxy-terminal protected derivative thereof covalently linked via a peptide bond to a peptide substrate comprising aspartate that is specifically cleaved by a protease specific for the substrate.

    摘要翻译： 提供了一种敏感的生物发光测定法，用于检测蛋白酶，包括特异性切割具有天冬氨酸的肽底物的胱天蛋白酶和蛋白酶。 在一个实施方案中，该测定使用通过肽键共价连接的氨基荧光素或其羧基末端保护的衍生物与底物共价连接到通过肽键与肽底物共价连接的胱天蛋白酶或氨基荧光素或其羧基端保护的衍生物 其包含特异性用于底物的蛋白酶特异性切割的天冬氨酸。

    Utility management system
    6.
    发明申请
    Utility management system 审中-公开
    实用管理系统

    公开(公告)号：US20050149345A1

    公开(公告)日：2005-07-07

    申请号：US10751759

    申请日：2004-01-05

    申请人： William Daily

    发明人： William Daily

    IPC分类号： G01D4/00 ,  G06Q30/00 ,  G06F17/60

    CPC分类号： G06Q30/04 ,  G01D4/004 ,  Y02B90/242 ,  Y04S20/322

    摘要： A remote site measures utility data and pushes the data to a processing site. The processing site processes the utility data into a utility usage report that takes into account customer desired spreadsheets, setup parameters, and utility tariff data. The usage report is pushed to the customer or other recipient in the form of an email or other communication.

    摘要翻译： 远程站点测量实用程序数据并将数据推送到处理站点。 处理站点将实用程序数据处理到实用程序使用情况报告中，该报告考虑到客户期望的电子表格，设置参数和实用程序资费数据。 使用报告以电子邮件或其他通信的形式推送给客户或其他收件人。

    LUMINESCENCE-BASED METHODS AND PROBES FOR MEASURING CYTOCHROME P450 ACTIVITY
    7.
    发明申请
    LUMINESCENCE-BASED METHODS AND PROBES FOR MEASURING CYTOCHROME P450 ACTIVITY 有权
    用于测量细胞色素P450活性的基于荧光的方法和探针

    公开(公告)号：US20110003316A1

    公开(公告)日：2011-01-06

    申请号：US12754164

    申请日：2010-04-05

    申请人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel Kin Sang Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

    发明人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel Kin Sang Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

    IPC分类号： G01N33/53 ,  C12Q1/66

    CPC分类号： G01N33/502 ,  A61K49/0008 ,  C07D417/04 ,  C12Q1/66 ,  G01N33/5088 ,  G01N33/582 ,  G01N2333/90245 ,  G01N2333/90251 ,  G01N2500/10

    摘要： The present invention provides methods, compositions, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferin or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. Upon addition of the luciferin derivative or other luminogenic molecule into a P450 reaction, the P450 enzyme metabolizes the molecule into a bioluminescent enzyme substrate, e.g., luciferin and/or luciferin derivative metabolite, in a P450 reaction. The resulting metabolite(s) serves as a substrate of the bioluminescent enzyme, e.g., luciferase, in a second light-generating reaction. Luminescent cytochrome P450 assays with low background signals and high sensitivity are disclosed and isoform selectivity is demonstrated. The present invention also provides an improved method for performing luciferase reactions which employs added pyrophosphatase to remove inorganic pyrophosphate, a luciferase inhibitor which may be present in the reaction mixture as a contaminant or may be generated during the reaction. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible luciferase inhibitors.

    摘要翻译： 本发明提供了可用于分析细胞，组织和动物中的代谢活性以及筛选测试化合物对细胞色素P450活性的影响的方法，组合物，底物和试剂盒。 特别地，使用发光分子的一步和两步法。 荧光素或coelenterazines，其是细胞色素P450底物并且也是生物发光酶，例如萤光素酶，原底物。 在P450反应中加入荧光素衍生物或其他发光分子时，P450酶在P450反应中将分子代谢为生物发光酶底物，例如荧光素和/或荧光素衍生物代谢产物。 所产生的代谢物在第二次发光反应中用作生物发光酶，例如荧光素酶的底物。 公开了具有低背景信号和高灵敏度的发光细胞色素P450测定，证明了同工型选择性。 本发明还提供一种用于进行荧光素酶反应的改进方法，其使用添加的焦磷酸酶来除去无机焦磷酸盐，荧光素酶抑制剂，其可作为污染物存在于反应混合物中，或可能在反应期间产生。 本发明方法还提供了一种使用荧光素酶稳定剂如可逆荧光素酶抑制剂在基于荧光素酶的测定中稳定和延长发光信号的方法。

    METHODS TO QUENCH LIGHT FROM OPTICAL REACTIONS
    8.
    发明申请
    METHODS TO QUENCH LIGHT FROM OPTICAL REACTIONS 审中-公开
    从光学反应中排除光的方法

    公开(公告)号：US20080248511A1

    公开(公告)日：2008-10-09

    申请号：US12056073

    申请日：2008-03-26

    申请人： William Daily ,  Erika Hawkins ,  Dieter Klaubert ,  Mark McDougall ,  James Unch ,  Keith V. Wood ,  Wenhui Zhou ,  Ji Zhu

    发明人： William Daily ,  Erika Hawkins ,  Dieter Klaubert ,  Mark McDougall ,  James Unch ,  Keith V. Wood ,  Wenhui Zhou ,  Ji Zhu

    IPC分类号： C07D209/16 ,  C12Q1/66

    CPC分类号： C12Q1/66

    摘要： The present invention relates to single and dual reporter luminescence assays utilizing reagents to quench an optical, e.g., an enzyme-mediated luminescence, reaction. In one embodiment of the invention, a reagent is added to an assay which selectively quenches a first enzyme-mediated luminescence reaction without affecting a subsequent distinct enzyme-mediated luminescent reaction(s). An assay kit containing one or more selective quench reagents, and compositions comprising the quench reagent(s), are also provided.

    摘要翻译： 本发明涉及使用试剂淬灭光学例如酶介导的发光反应的单和双重报道发光测定。 在本发明的一个实施方案中，将试剂加入到选择性淬灭第一酶介导的发光反应而不影响随后不同的酶介导的发光反应的测定中。 还提供了含有一种或多种选择性骤冷试剂和包含骤冷试剂的组合物的测定试剂盒。