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    Electrophysiological assay using oocytes that express human ENaC and the use phenamil to improve the effect of ENaC enhancers in assays using membrane potential reporting dyes
    1.
    发明申请
    Electrophysiological assay using oocytes that express human ENaC and the use phenamil to improve the effect of ENaC enhancers in assays using membrane potential reporting dyes 有权
    使用表达人类ENaC的卵母细胞进行电生理测定，并使用苯胺胺改善ENaC增强子在使用膜电位报告染料的测定中的作用

    公开(公告)号：US20050059094A1

    公开(公告)日：2005-03-17

    申请号：US10887233

    申请日：2004-07-09

    申请人： Guy Servant ,  Hong Chang ,  Cyril Redcrow ,  Sumita Ray ,  Imran Clark

    发明人： Guy Servant ,  Hong Chang ,  Cyril Redcrow ,  Sumita Ray ,  Imran Clark

    IPC分类号： C07H21/04 ,  C07K1/00 ,  C12N1/20 ,  C12P21/06 ,  C12Q20060101 ,  G01N33/50 ,  G01N33/53 ,  G01N33/566 ,  G01N33/567 ,  G01N33/68

    CPC分类号： G01N33/6872 ,  C12N15/85 ,  C12N2320/10 ,  C12N2503/02 ,  C12Q1/025 ,  G01N33/5044 ,  G01N2500/10

    摘要： In one aspect, the present invention relates to a mammalian cell-based high-throughput assay for the profiling and screening of human epithelial sodium channel (hENaC) cloned from a human kidney c-DNA library and is also expressed in other tissues including human taste tissue. The present invention further relates to amphibian oocyte-based medium-throughput electrophysiological assays for identifying human ENaC modulators, preferably ENaC enhancers. Compounds that modulate ENaC function in a cell-based ENaC assay are expected to affect salty taste in humans. The assays described herein have advantages over existing cellular expression systems. In the case of mammalian cells, such assays can be run in standard 96 or 384 well culture plates in high-throughput mode with enhanced assay results being achieved by the use of a compound that inhibits ENaC function, preferably an amiloride derivative such as Phenamil. In the case of the inventive oocyte electrophysiological assays (two-electrode voltage-clamp technique), these assays facilitate the identification of compounds which specifically modulate human ENaC. The assays of the invention provide a robust screen useful to detect compounds that facilitate (enhance) or inhibit hENaC function. Compounds that enhance or block human ENaC channel activity should thereby modulate salty taste in humans.

    摘要翻译： 一方面，本发明涉及从人肾C-DNA文库克隆的人上皮钠通道（hENaC）的分析和筛选的基于哺乳动物细胞的高通量测定，并且还在包括人类味道的其它组织中表达 组织。 本发明还涉及用于鉴定人类ENaC调节剂，优选ENaC增强子的两栖动物基于卵母细胞的中等通量电生理测定法。 在基于细胞的ENaC测定中调节ENaC功能的化合物预期会影响人类的咸味。 本文所述的测定法优于现有的细胞表达系统。 在哺乳动物细胞的情况下，这种测定可以在高通量模式的标准96或384孔培养板中进行，通过使用抑制ENaC功能的化合物，优选阿米洛利衍生物，如苯胺，可以实现增强的测定结果。 在本发明的卵母细胞电生理测定（双电极电压钳技术）的情况下，这些测定有助于鉴定特异性调节人类ENaC的化合物。 本发明的测定提供了可用于检测促进（增强）或抑制hENaC功能的化合物的强壮屏幕。 增强或阻断人类ENaC通道活性的化合物应该调节人体的咸味。

    Electrophysiological assays using oocytes that express human enac and the use of phenamil to improve the effect of enac enhancers in assays using membrane potential reporting dyes
    2.
    发明申请
    Electrophysiological assays using oocytes that express human enac and the use of phenamil to improve the effect of enac enhancers in assays using membrane potential reporting dyes 审中-公开
    使用表达人类烯醇的卵母细胞进行电生理测定，以及使用苯胺胺改善enac增强子在使用膜电位报告染料的测定中的作用

    公开(公告)号：US20090123942A1

    公开(公告)日：2009-05-14

    申请号：US10563758

    申请日：2004-07-09

    申请人： Guy Servant ,  Hong Chang ,  Cyril Redcrow ,  Sumita Ray ,  Imran Clark ,  Bryan Moyer

    发明人： Guy Servant ,  Hong Chang ,  Cyril Redcrow ,  Sumita Ray ,  Imran Clark ,  Bryan Moyer

    IPC分类号： C12Q1/02 ,  G01N33/554 ,  C12N5/06

    CPC分类号： G01N33/6872 ,  C12N15/85 ,  C12N2320/10 ,  C12N2503/02 ,  C12Q1/025 ,  G01N33/5044 ,  G01N2500/10

    摘要： In one aspect, the present invention relates to a mammalian cell-based high-throughput assay for the profiling and screening of human epithelial sodium channel (hENaC) cloned from a human kidney c-DNA library and is also expressed in other tissues including human taste tissue. The present invention further relates to amphibian oocyte-based medium-throughput electrophysiological assays for identifying human ENaC modulators, preferably ENaC enhancers. Compounds that modulate ENaC function in a cell-based ENaC assay are expected to affect salty taste in humans. The assays described herein have advantages over existing cellular expression systems. In the case of mammalian cells, such assays can be run in standard 96 or 384 well culture plates in high-throughput mode with enhanced assay results being achieved by the use of a compound that inhibits ENaC function, preferably an amiloride derivative such as Phenamil. In the case of the inventive oocyte electrophysiological assays (two-electrode voltage-clamp technique), these assays facilitate the identification of compounds which specifically modulate human ENaC. The assays of the invention provide a robust screen useful to detect compounds that facilitate (enhance) or inhibit hENaC function. Compounds that enhance or block human ENaC channel activity should thereby modulate salty taste in humans.

    摘要翻译： 一方面，本发明涉及从人肾C-DNA文库克隆的人上皮钠通道（hENaC）的分析和筛选的基于哺乳动物细胞的高通量测定，并且还在包括人类味道的其它组织中表达 组织。 本发明还涉及用于鉴定人类ENaC调节剂，优选ENaC增强子的两栖动物基于卵母细胞的中等通量电生理测定法。 在基于细胞的ENaC测定中调节ENaC功能的化合物预期会影响人类的咸味。 本文所述的测定法优于现有的细胞表达系统。 在哺乳动物细胞的情况下，这种测定可以在高通量模式的标准96或384孔培养板中进行，通过使用抑制ENaC功能的化合物，优选阿米洛利衍生物，如苯胺，可以实现增强的测定结果。 在本发明的卵母细胞电生理测定（双电极电压钳技术）的情况下，这些测定有助于鉴定特异性调节人类ENaC的化合物。 本发明的测定提供了可用于检测促进（增强）或抑制hENaC功能的化合物的强壮屏幕。 增强或阻断人类ENaC通道活性的化合物应该调节人体的咸味。

    Method of modulating human ENaC sodium channel
    7.
    发明授权
    Method of modulating human ENaC sodium channel 有权
    调节人类ENaC钠通道的方法

    公开(公告)号：US08105792B2

    公开(公告)日：2012-01-31

    申请号：US10887233

    申请日：2004-07-09

    申请人： Guy Servant ,  Hong Chang ,  Cyril Redcrow ,  Sumita Ray ,  Imran Clark ,  Bryan Moyer

    发明人： Guy Servant ,  Hong Chang ,  Cyril Redcrow ,  Sumita Ray ,  Imran Clark ,  Bryan Moyer

    IPC分类号： G01N33/53

    CPC分类号： G01N33/6872 ,  C12N15/85 ,  C12N2320/10 ,  C12N2503/02 ,  C12Q1/025 ,  G01N33/5044 ,  G01N2500/10

    摘要： In one aspect, the present invention relates to a mammalian cell-based high-throughput assay for the profiling and screening of human epithelial sodium channel (hENaC) cloned from a human kidney c-DNA library and is also expressed in other tissues including human taste tissue. The present invention further relates to amphibian oocyte-based medium-throughput electrophysiological assays for identifying human ENaC modulators, preferably ENaC enhancers. Compounds that modulate ENaC function in a cell-based ENaC assay are expected to affect salty taste in humans. The assays described herein have advantages over existing cellular expression systems. In the case of mammalian cells, such assays can be run in standard 96 or 384 well culture plates in high-throughput mode with enhanced assay results being achieved by the use of a compound that inhibits ENaC function, preferably an amiloride derivative such as Phenamil. In the case of the inventive oocyte electrophysiological assays (two-electrode voltage-clamp technique), these assays facilitate the identification of compounds which specifically modulate human ENaC. The assays of the invention provide a robust screen useful to detect compounds that facilitate (enhance) or inhibit hENaC function. Compounds that enhance or block human ENaC channel activity should thereby modulate salty taste in humans.

    摘要翻译： 一方面，本发明涉及从人肾C-DNA文库克隆的人上皮钠通道（hENaC）的分析和筛选的基于哺乳动物细胞的高通量测定，并且还在包括人类味道的其它组织中表达 组织。 本发明还涉及用于鉴定人类ENaC调节剂，优选ENaC增强子的两栖动物基于卵母细胞的中等通量电生理测定法。 在基于细胞的ENaC测定中调节ENaC功能的化合物预期会影响人类的咸味。 本文所述的测定法优于现有的细胞表达系统。 在哺乳动物细胞的情况下，这种测定可以在高通量模式的标准96或384孔培养板中进行，通过使用抑制ENaC功能的化合物，优选阿米洛利衍生物，如苯胺，可以实现增强的测定结果。 在本发明的卵母细胞电生理测定（双电极电压钳技术）的情况下，这些测定有助于鉴定特异性调节人类ENaC的化合物。 本发明的测定提供了可用于检测促进（增强）或抑制hENaC功能的化合物的强壮屏幕。 增强或阻断人类ENaC通道活性的化合物应该调节人体的咸味。