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    • 7. 发明申请
    • ASSAYS FOR IDENTIFYING GENES ENCODING SALTY TASTE RECEPTORS AND MODULATORS
    • 用于识别编码盐类受体和调节剂的基因的测定
    • WO2010065863A3
    • 2010-10-14
    • PCT/US2009066788
    • 2009-12-04
    • SENOMYX INCMOYER BRYANZLOTNIK ALBERTHEVEZI PETERSOTO HORTENSIALU MINGAO NASAGANICH MICHAELDAHAN MICHAELLAITA BIANCASERVANT GUYBRUST PAULWILLIAMS MARK
    • MOYER BRYANZLOTNIK ALBERTHEVEZI PETERSOTO HORTENSIALU MINGAO NASAGANICH MICHAELDAHAN MICHAELLAITA BIANCASERVANT GUYBRUST PAULWILLIAMS MARK
    • C07K14/705
    • G01N33/5008A01K2267/03C07K14/705G01N2333/705
    • The present invention relates to rationales for selecting salty taste receptors from a discrete set of taste specific genes disclosed infra by use of methods including electrophysiology, bioiirformatics, and quantitative gene expression assays using transgenic animals as well as functional assays wherein these salty taste receptors may be expressed alone or in association with other taste specific polypeptides such as TRPML3, PKD2L1 or GPRl 13. In addition the invention relates to the use of the identified salty taste receptors in assays, preferably high- throughput, in order to identify modulators which maybe used as flavor additives to modulate salty taste. Preferably, the invention provides high- throughput mammalian and medium- throughput oocyte- based electrophysiological assays for identifying human TRPML3 modulators, preferably TRPML3 enhancers. Compounds that modulate TRPML3 function in the assay are expected to affect salty taste in humans. The inventive electrophysiological assays, such as the two-electrode voltage-clamp technique, facilitate the identification of compounds which specifically modulate human TRPML3. The assays of the invention provide a robust screen useful to detect compounds that facilitate (enhance) or inhibit TRPML3 function. Compounds that enhance or block TRPML3 channel activity should thereby modulate salty taste. In addition, these compounds maybe used to regulate sodium excretion, urinary output and other biological functions relating to sodium levels. This invention relates to the elucidation that TRPML3 is involved in salty taste perception in primates including humans and likely other mammals (given the significance of sodium and other ions to physiological functions and conditions this phenotype is likely strongly conserved in different animals). The TRPML3 gene also modulates one or more of sodium metabolism, sodium excretion, blood pressure, fluid retention, cardiac function and urinary functions such as urine production and excretion.
    • 本发明涉及通过使用包括电生理学,生物信息学和使用转基因动物的定量基因表达测定的方法以及其中这些咸味受体可能的功能测定法从下文所公开的离散的味道特异性基因组中选择咸味受体的理由 单独表达或与其它味道特异性多肽如TRPML3,PKD2L1或GPR113共同表达。此外,本发明涉及鉴定的咸味受体在测定中的应用,优选高通量,以便鉴定可用作 风味添加剂调味咸味。 优选地,本发明提供用于鉴定人类TRPML3调节剂(优选TRPML3增强子)的高通量哺乳动物和中等通量的基于卵母细胞的电生理测定。 在测定中调节TRPML3功能的化合物预期会影响人体的咸味。 本发明的电生理测定,例如双电极电压钳技术,有助于鉴定特异性调节人TRPML3的化​​合物。 本发明的测定法提供了一种可用于检测促进(增强)或抑制TRPML3功能的化合物的稳健筛选。 增强或阻断TRPML3通道活性的化合物应该调节咸味。 此外,这些化合物也可用于调节与钠水平相关的钠排泄,泌尿生产和其他生物学功能。 本发明涉及TRPML3涉及包括人和可能的其他哺乳动物在内的灵长类动物的咸味感知(给出钠和其它离子对生理功能和条件的重要性,该表型可能在不同动物中强烈保守)。 TRPML3基因还调节钠代谢,钠排泄,血压,流体保留,心脏功能和尿液功能如尿液产生和排泄中的一种或多种。