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1. 发明申请

US20060134693A1 Olfactory cyclic nucleotide-gated channel cell-based assays to identify T1R and T2R taste modulators 审中-公开
标题翻译：基于嗅觉环状核苷酸门控通道细胞的测定来鉴定T1R和T2R调味剂
公开(公告)号：US20060134693A1
公开(公告)日：2006-06-22
申请号：US11006802
申请日：2004-12-08
申请人： Guy Servant , Mark Ozeck , Paul Brust , Hong Xu , Lena Staszewski , Jon Adler , Mark Zoller , Nicholas Callamaras , Alexey Pronin , Bryan Moyer
发明人： Guy Servant , Mark Ozeck , Paul Brust , Hong Xu , Lena Staszewski , Jon Adler , Mark Zoller , Nicholas Callamaras , Alexey Pronin , Bryan Moyer
IPC分类号： G01N33/53 , C07K14/705
CPC分类号： C07K14/705 , G01N33/5008 , G01N33/502 , G01N33/566 , G01N2333/726
摘要： Screening assays, preferably high throughput, are provided that screen libraries of candidate compounds to identify agonists, antagonists, enhancers or modulators of taste receptors (bitter, sweet or savory (umami) taste receptor) using test cells that co-express at least one functional taste receptor and an olfactory cyclic nucleotide-gated channel (oCNGC). (The oCNGC preferably comprises at least one mutation in one or more subunits that renders the resultant oCNGC more sensitive to CAMP (which in turn enhances the sensitivity of assay using this oCNGC). These taste modulatory compounds are identified based on their effect on oCNGC activity, e.g., using fluorimetric assays that screen for changes in intracellular calcium or sodium concentration in test cells that co-express at least one taste receptor, oCNGC and a Gαi/o protein.
摘要翻译：提供筛选分析，优选高通量，使用候选化合物的筛选文库，以鉴定共感染至少一种功能性的测试细胞的味觉受体（苦味，甜味或咸味（鲜味）味觉受体）的激动剂，拮抗剂，增强剂或调节剂味觉受体和嗅觉环状核苷酸门控通道（oCNGC）。（oCNGC优选地包含一个或多个亚基中的至少一个突变，使得所得到的oCNGC对CAMP更敏感（反过来又增强了使用该oCNGC的测定的灵敏度）。这些口味调节化合物基于它们对oCNGC活性的影响例如，使用荧光测定法筛选测试细胞中细胞内钙或钠浓度的变化，所述测试细胞共表达至少一种味觉受体（oCNGC）和蛋白质（G）。

2. 发明申请

US20070037134A1 Functional coupling of T1Rs and T2Rs by Gi proteins, and cell-based assays for the identification of T1R and T2R modulators 审中-公开
标题翻译：通过Gi蛋白的T1R和T2Rs的功能偶联和用于鉴定T1R和T2R调节剂的基于细胞的测定
公开(公告)号：US20070037134A1
公开(公告)日：2007-02-15
申请号：US11395375
申请日：2006-04-03
申请人： Guy Servant , Mark Ozeck , Paul Brust , Hong Xu
发明人： Guy Servant , Mark Ozeck , Paul Brust , Hong Xu
IPC分类号： G01N33/567 , C12Q1/00
CPC分类号： G01N33/5008 , G01N33/5041 , G01N33/566 , G01N2333/726 , G01N2500/10
摘要： The invention resides in part in the discovery that G proteins other than Gα15 couples to T1R and T2R taste receptors, particularly Gi proteins such as Gαi. Related to this discovery, the invention provides cell-based assay methods for identifying compounds that modulate the activity of specific T1R or T2R taste receptors or which modulate the effect of other T1R or T2R modulators on T1R or T2R activity. These assay methods preferably detect the effect of a putative T1R or T2R modulator compound on MAPK activation cAMP accumulation, or adenylyl cyclase activity or another signaling pathway regulated by Gi proteins. The level of MAPK activation, cAMP accumulation or adenylyl cyclase is preferably determined by immunoassay methods that use ligands (monoclonal or polyclonal antibodies) that specifically bind an activated (phosphorylated) MAPK, cAMP, or adenylyl cyclase.
摘要翻译：本发明部分地在于以下发现：除了Galpha 15之外的G蛋白与T1R和T2R味觉受体，特别是GII蛋白（例如Gα- SUB>。与该发现有关，本发明提供用于鉴定调节特异性T1R或T2R味觉受体的活性的化合物的基于细胞的测定方法，或调节其它T1R或T2R调节剂对T1R或T2R活性的影响。这些测定方法优选检测推测的T1R或T2R调节剂化合物对MAPK激活cAMP积聚或腺苷酸环化酶活性或由蛋白质调节的另一信号传导途径的影响。 MAPK活化水平，cAMP积聚或腺苷酸环化酶优选通过使用特异性结合活化（磷酸化）MAPK，cAMP或腺苷酸环化酶的配体（单克隆或多克隆抗体）的免疫测定方法来确定。

3. 发明授权

US07022488B2 Functional coupling of T1Rs and T2Rs by Gi proteins, and cells-based assays for the identification of T1R and T2R modulators 有权
标题翻译：通过Gi蛋白的T1R和T2Rs的功能偶联，以及用于鉴定T1R和T2R调节剂的基于细胞的测定
公开(公告)号：US07022488B2
公开(公告)日：2006-04-04
申请号：US10770127
申请日：2004-02-03
申请人： Guy Servant , Mark Ozeck , Paul Brust , Hong Xu
发明人： Guy Servant , Mark Ozeck , Paul Brust , Hong Xu
IPC分类号： G01N33/53 , G01N33/567 , C12P21/06 , C12N1/20
CPC分类号： G01N33/5041 , G01N33/5008 , G01N33/566 , G01N2333/726 , G01N2500/10
摘要： The invention resides in part in the discovery that G proteins other than Gα15 couples to T1R and T2R taste receptors, particularly Gi proteins such as Gαi. Related to this discovery, the invention provides cell-based assay methods for identifying compounds that modulate the activity of specific T1R or T2R taste receptors or which modulate the effect of other T1R or T2R modulators on T1R or T2R activity. These assay methods preferably detect the effect of a putative T1R or T2R modulator compound on MAPK activation, cAMP accumulation, or adenylyl cyclase activity or another signaling pathway regulated by Gi proteins. The level of MAPK activation, cAMP accumulation or adenylyl cyclase is preferably determined by immunoassay methods that use ligands (monoclonal or polyclonal antibodies) that specifically bind an activated (phosphorylated) MAPK, cAMP, or adenylyl cyclase.
摘要翻译：本发明部分地在于以下发现：除了Galpha 15之外的G蛋白与T1R和T2R味觉受体，特别是GII蛋白（例如Gα- SUB>。与该发现有关，本发明提供用于鉴定调节特异性T1R或T2R味觉受体的活性的化合物的基于细胞的测定方法，或调节其它T1R或T2R调节剂对T1R或T2R活性的影响。这些测定方法优选地检测推定的T1R或T2R调节剂化合物对MAPK激活，cAMP积聚或腺苷酸环化酶活性或由GII蛋白调节的另一信号传导途径的影响。 MAPK活化水平，cAMP积聚或腺苷酸环化酶优选通过使用特异性结合活化（磷酸化）MAPK，cAMP或腺苷酸环化酶的配体（单克隆或多克隆抗体）的免疫测定方法来确定。

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